WO2006074227A2 - HIGHLY ANTIPROLIFERATIVE, LOW-CALCEMIC, ANALOGS OF THE HORMONE 1α, 25-DIHYDROXYVITAMIN D3 WITH CARBONYL SIDE CHAINS - Google Patents

Abstract

The present invention discloses a series of new carbonyl side chain anlogs of calcitriol (I). Unexpectedly, several of these ketone analogs, such as the 24- and 25-tert-butyl ketones, amides, hydroxymate esters and a hetero atom ketones even though lacking the classical side-chain tertiary hydroxyl group, are considerably more antiproliferative in vitro than the hormone calcitriol (I) even at physiologically relevant low nanomolar concentrations and are less calcemic than calcitriol (I) in vivo.

Description

HIGHLY ANTIPROLIFERATIVE, LOW-CALCEMIC, ANALOGS OF THE HORMONE lα, 25-DIHYDROXYVITAMIN D₃ WITH CARBONYL

SIDE CHAINS

CROSS-REFERENCES TO RELATED APPLICATIONS

[0001] This application claims the benefit of United States Provisional Patent Application No. 60/641,340, filed January 4, 2005, which is incorporated herein by reference in its entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER

FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This study was supported in-part by National Institutes of Health grant CA 93547 (to G.H.P.) and all joint inventors have assigned their rights to Johns Hopkins University.

BACKGROUND OF THE INVENTION

[0003] For chemotherapy of various human illnesses, medicinal organic chemists are designing and preparing analogs that are at least as potent as, but less calcemic than, the natural hormone lα,25-dihydroxyvitamin D3 (I, calcitriol), Holick, M.F., VitaminD: Molecular Biology, Physiology, and Clinical Applications; Humana Press: Totowa, NJ, 1999; Feldman, D.; et al, Vitamin D; Academic Press: San Diego, CA, 1997; Bouillon, R.; et al, Endocr. Rev., 16: 200-257 (1995); Posner, G. H., et al., Eur. J. Org. Chem., 20:3889-3895 (2003).

[0004] Currently, approximately eight such analogs of calcitriol (I) are regularly used as drugs that promote healthier living, (Posner, G. H., et al, Eur. J. Org. Chem., 20:3889- 3895 (2003)). A continuing chemical challenge, however, is to determine which small structural changes and which simple functional group alterations in the large natural hormone molecule will elicit desirable biological responses.

[0005] Some side-chain sulfone (Posner, G. H., et al., J. Steroid Biochem. MoI. Biol, 89- 90:5-12 (2004) and sulfoximine (Kahraman, M., et al, H. J. Med. Chem., 47:6854-6863 (2004) analogs have been designed and prepared such that these functional groups might act as H-bond acceptors (rather than as H-bond donors like the natural 25-OH group) toward the vitamin D receptor (VDR); several of these new analogs are potent, selective, and low-calcemic inhibitors of the human CYP24 hydroxylase enzyme. (Posner, G. H., et al, J. Steroid Biochem. MoI Biol, 89-90:5-12 (2004); and Kahraman, M., et al, H. J. Med. Chem., 47:6854-6863 (2004)). Hoffman-La Roche researchers showed that incorporating a 16,17-double bond into an analog generally promotes antiproliferative activity (Uskokovic, M. R., et al, Curr. Pharm. Des., 3:99-123 (1997)). Reddy and colleagues (Siu-Caldera, M., et al, J. Steroid Biochem. MoI. Biol, 59:405-412 (1996)) showed that such a 16-ene 24-ketone 25-OH metabolite is even more antiproliferatively potent than its 24-CH₂ non-ketone parent, and we have similarly observed the high antiproliferative activity of a 16-ene 25-ketone analog (Posner, G. H., et al, J. Med. Chem., 45:1723-1730 (2002)). Finally, DeLuca and colleagues (Plum, L. A., et al, Proc. Natl. Acad. ScL U.S.A., 101:6900-6904 (2004)) have reported recently a biologically active abbreviated side-chain analog having no side-chain heteroatom. [0006] Understanding such chemical structure-biological activity relationships (SAR) is important for rational design of new analogs as potential drug candidates, Bouillon, R., et al, Endocr. Rev., 16:200-257 (1995) and Posner, G. H., et al, Eur. J. Org. Chem., 20:3889-3895 (2003).

BRIEF SUMMARY OF THE INVENTION

[0007] hi one aspect of the present invention an antiproliferative, low-calcemic compound having the Formula (II):

wherein L is selected from the following side chains:

wherein n = 1, 2;

m = 0-2;

p - 3-6;

Z = CH₂, O, NR¹, S₃ SO, SO₂, O-N-R¹, N(R')-N(R^?), -(C=O)-;

Y = O₅CH₂;

X = C, O, S₅ N;

V = H₅ Or V = V = CH₂;

R = isoalkyl, t-butyl, l-(Methyl)cycloalkyl (3-7), l-(OH)cycloalkyl (3-7), 1-adamantyl;

R¹ = H₅ F₅ Me, CD₃, Et₅ z-Pr, allyl, CH₂OR¹, CMe₂Ph, R=R'=cycloalkyl (3-7); and

R" = H, F, Me₅ CD₃, Et, z-Pr, allyl, CH₂OR', R'=R'=cycloalkyl (3-7),

is disclosed.

[0008] In another aspect, the present invention provides methods of treating a disorder or condition such as but not limited to osteoporesis, psoriasis, immunosuppressed states. The method includes administering to a subject in need of such treatment, an effective amount of the compound of Formula (II).

[0009] In another aspect, the present invention provides a pharmaceutical composition. The pharmaceutical composition includes a pharmaceutically acceptable excipient and a compound having the Formula (II):

wherein L is selected from the following side chains:

wherein n = 1, 2;

m = 0-2;

P - 3-6;

Z = CH₂, O, NR', S, SO, SO₂, O-N-R¹, N(R')-N(R'), -(C=O)-;

Y = O₅CH₂;

X = C, O, S, N;

V = H₅ Or V = V = CH₂;

R = isoalkyl, t-butyl, l-(Methyl)cycloalkyl (3-7), l-(OH)cycloalkyl (3-7),l-adamantyl; R¹ = H, F, Me, CD₃, Et, z-Pr, allyl, CH₂OR¹, CMe₂Ph, R^R^cycloalkyl (3-7); and

R" = H₅ F, Me, CD₃, Et₃ /-Pr, allyl, CH₂OR¹, R'=R'=cycloalkyl (3-7),

is disclosed.

[0010] Toward this goal, a series of calcitriol analogs, have been developed, that lack the natural 25-OH group widely thought to be necessary for high biological activity (Holick, M.F., VitaminD: Molecular Biology, Physiology, and Clinical Applications; Humana Press: Totowa, NJ, 1999; Feldman, D.; et al, Vitamin D; Academic Press: San Diego, CA, 1997; Bouillon, R.; et al, Endocr. Rev., 16: 200-257 (1995)). In addition to these ketones the present invention is directed toward highly antiproliferative, low-calcemic analogs with carbonyl side chains such as amides, hydroxymate esters and αheteroatom ketones.

BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The accompanying drawings, which are incorporated in and form a part of the specification, illustrate the preferred embodiments of the present invention, and together with the description serve to explain the principles of the invention.

[0012] In the drawings, Figures 1-10, the horizontal axis depicts various dilutions of the test compounds, ranging from 10^"6 to 10^"9 molar, that were exposed to murine keratinocytes. The vertical axis (cell number) depicts the number of murine keratinocyte cells present after exposure to a specific concentration of the tested compound as compared to the cell number at time zero.

[0013] hi the drawings, Figures 11-15, the effects of the test compounds on calcium levels in rat urine are measured. The horizontal axis depicts various dilutions of the test compound, ranging from 1 μg/Kg to 50 /xg/Kg, that were exposed to the test animals. The vertical axis depicts the quantity of calcium excreted per Kg body weight when exposed to a specific concentration of the tested compound as compared to the quantity of calcium excreted per Kg body weight when exposed only to the vehicle and calcitrol. In the Drawings:

[0014] Figure 1 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compounds Ia, 4a, 3 and 5 of the present invention versus calcitriol (I) and versus a control using only a solvent. [0015] Figure 2 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compound 2 of the present invention versus calcitriol (I) and versus a control using only a solvent.

[0016] Figure 3 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compounds 5, 6 and 9 of the present invention versus calcitriol (I) and versus a control using only a solvent.

[0017] Figure 4 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compounds 3, 7 and 11 of the present invention versus calcitriol (I) and versus a control using only a solvent. [0018] Figure 5 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compounds 10 and 14 of the present invention versus calcitriol (I) and versus a control using only a solvent.

[0019] Figure 6 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compounds Ia and 12 of the present invention versus calcitriol (I) and versus a control using only a solvent.

[0020] Figure 7 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compounds 4a, 12 and 15 of the present invention versus calcitriol (I) and versus a control using only a solvent.

[0021] Figure 8 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compound 16 of the present invention versus calcitriol (I) and versus a control using only a solvent.

[0022] Figure 9 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compound 17 of the present invention versus calcitriol (I) and versus a control using only a solvent. [0023] Figure 10 depicts the dose response curves generated by exposing murine keratinocytes to various concentrations of compound 20 of the present invention versus calcitriol (I) and versus a control using only a solvent.

[0024] Figure 11 depicts the effect of compounds Ia, 2, 4a and 3 on calcium levels in rat urine versus calcitriol (I) and versus a control using only a solvent. [0025] Figure 12 depicts the effect of compounds Ia, and 2 on calcium levels in rat urine versus calcitriol (I) and versus a control using only a solvent.

[0026] Figure 13 depicts the effect of compounds 3 and 11 on calcium levels in rat urine versus calcitriol (I) and versus a control using only a solvent. [0027] Figure 14 depicts the effect of compounds 4a and 12 on calcium levels in rat urine versus calcitriol (I) and versus a control using only a solvent.

[0028] Figure 15 depicts the effect of compounds 16 and 20 on calcium levels in rat urine versus calcitriol (I) and versus a control using only a solvent. [0029] Figure 16 schematically depicts the synthesis scheme for compounds Ia and Ib of the present invention.

[0030] Figure 17 schematically depicts the synthesis scheme for compound 2 of the present invention.

[0031] Figure 18 schematically depicts the synthesis scheme for compound 3 of the present invention.

[0032] Figure 19 schematically depicts the synthesis scheme for compounds 4a and 4b of the present invention.

[0033] Figure 20 schematically depicts the synthesis scheme for compound 5 of the present invention. [0034] Figure 21 schematically depicts the synthesis scheme for compounds 6 and 7 of the present invention.

[0035] Figure 22 schematically depicts the synthesis scheme for compound 8 of the present invention.

[0036] Figure 23 schematically depicts the synthesis scheme for compound 9 of the present invention.

[0037] Figure 24 schematically depicts the synthesis scheme for compound 10 of the present invention.

[0038] Figure 25 schematically depicts the synthesis scheme for compounds 11 and 12 of the present invention. [0039] Figure 26 schematically depicts the synthesis scheme for compounds 13 and 13a of the present invention.

[0040] Figure 27 schematically depicts the synthesis scheme for compound 14 of the present invention.

[0041] Figure 28 schematically depicts the synthesis scheme for compound 15 of the present invention.

[0042] Figure 29 schematically depicts the synthesis scheme for compound 16 of the present invention.

[0043] Figure 30 schematically depicts the synthesis scheme for compound 17 of the present invention. [0044] Figure 31 schematically depicts the synthesis scheme for compound 18 of the present invention.

[0045] Figure 32 schematically depicts the synthesis scheme for compound 19 of the present invention. [0046] Figure 33 schematically depicts the synthesis scheme for compound 20 of the present invention. [0047] Figure 34 depicts the generic structure of the compounds of the present invention.

DETAILED DESCRIPTION OF THE INVENTION ABBREVIATIONS AND DEFINITIONS

[0048] The abbreviations used herein have their conventional meaning within the chemical and biological arts.

[0049] Where moieties are specified by their conventional chemical formulae, written from left to right, they equally encompass the chemically identical moieties that would result from writing the structure from right to left, e.g., -CH₂O- is equivalent to -OCH₂-.

[0050] The term "alkyl," by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e. unbranched) or branched chain, or cyclic hydrocarbon radical, or combination thereof, which may be fully saturated, mono- or polyunsaturated and can include di- and multivalent radicals, having the number of carbon atoms designated (i.e. C₁-C₁₀ means one to ten carbons). Examples of saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group is one having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2- isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(l,4-pentadienyl), ethynyl, 1- and 3- propynyl, 3-butynyl, and the higher homologs and isomers.

[0051] The term "alkylene" by itself or as part of another substituent means a divalent radical derived from an alkyl, as exemplified, but not limited, by -CH₂CH₂CH₂CH₂-. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, including those groups having 10 or fewer carbon atoms. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms. [0052] The terms "alkoxy," "alkylamino," and "alkylthio" (or thioalkoxy) are used in their conventional sense, and refer to those alkyl groups attached to the remainder of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively.

[0053] The term "heteroalkyl," by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain, or cyclic hydrocarbon radical, or combinations thereof, consisting of the stated number of carbon atoms and a heteroatom selected from the group consisting of O, N, P, Si and S, and wherein the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N, P and S and Si may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Examples include, but are not limited to, -CH₂-CH₂-O-CH₃, -CH₂-CH₂-NH-CH₃, -CH₂-CH₂-N(CHa)-CH₃, -CH₂-S-CH₂-CH₃, -CH₂-CH₂₃-S(O)-CH₃, -CH₂-CH₂-S(O)₂-CH₃, -CH=CH-O-CH₃, -Si(CH₃)S, -CH₂-CH=N-OCH₃, -CH=CH-N(CH₃)- CH₃, 0-CH₃, -0-CH₂-CH₃, and -CN. Up to two heteroatoms may be consecutive, such as, for example, -CH₂-NH-OCH₃ and -CH₂-O-Si(CH₃)₃. Similarly, the term "heteroalkylene" by itself or as part of another substituent means a divalent radical derived from heteroalkyl, as exemplified, but not limited by, -CH₂-CH₂-S-CH₂-CH₂- and -CH₂-S-CH₂-CH₂-NH-CH₂- . For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Still further, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula -C(O)₂R'- represents both -C(O)₂R'- and -R¹C(O)₂-. As described above, heteroalkyl groups, as used herein, include those groups that are attached to the remainder of the molecule through a heteroatom, such as -C(O)R', -C(O)NR', -NR'R", -OR', -SR', and/or -SO₂R'. Where "heteroalkyl" is recited, followed by recitations of specific heteroalkyl groups, such as -NR'R or the like, it will be understood that the terms heteroalkyl and -NR'R" are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited to add clarity. Thus, the term "heteroalkyl" should not be interpreted herein as excluding specific heteroalkyl groups, such as -NR'R or the like.

[0054] The terms "cycloalkyl" and "heterocycloalkyl", by themselves or in combination with other terms, represent, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl", respectively. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3- cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1 -(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4- morpholinyl, 3-morpholinyl, tefrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2- yl, tetrahydrothien-3-yl, 1 -piperazinyl, 2-piperazinyl, and the like.

[0055] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C₁-C₄)alkyl" is mean to include, but not be limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0056] The term "aryl" means, unless otherwise stated, a polyunsaturated, aromatic, hydrocarbon substituent which can be a single ring or multiple rings (preferably from 1 to 3 rings) which are fused together or linked covalently. The term "heteroaryl" refers to aryl groups (or rings) that contain from one to four heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3- pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4- oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5- thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5- isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituent moieties for each of the above noted aryl and heteroaryl ring systems may be selected from the group of acceptable substituent moieties described below.

[0057] For brevity, the term "aryl" when used in combination with other terms (e.g. , aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroaryl rings as defined above. Thus, the term "arylalkyl" is meant to include those radicals in which an aryl group is attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl and the like) including those alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced by, for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(l- naphthyloxy)propyl, and the like). [0058] The term "oxo" as used herein means an oxygen that is double bonded to a carbon atom.

[0059] Each of the above terms (e.g., "alkyl," "heteroalkyl," "aryl" and "lieteroaryl") are meant to include both substituted and unsubstituted forms of the indicated radical. Preferred substituent moieties for each type of radical are provided below.

[0060] Substituent moieties for the alkyl and heteroalkyl radicals (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one or more of a variety of groups selected from, but not limited to: -OR', =0, =NR', =N-0R', -NR¹R", -SR', -halogen, -SiR¹R¹¹R"¹, -OC(O)R¹, -C(O)R¹, -CO₂R', -CONR¹R", -OC(O)NR¹R", -NR¹¹C(O)R¹,

-NR'-C(O)NR"R"', -NR¹¹C(O)₂R¹, -NR-C(NR¹R¹¹R¹¹O=NR"", -NR-C(NR¹R¹O=NR"¹, -S(O)R¹, -S(O)₂R¹, -S(O)₂NR¹R", -NRSO₂R¹, -CN and -NO₂ in a number ranging from zero to (2m'+l), where m' is the total number of carbon atoms in such radical. R', R", R"¹ and R"" each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted alkyl, alkoxy or thioalkoxy groups, or arylalkyl groups. When a compound of the invention includes more than one R group, for example, each of the R groups is independently selected as are each R', R", R"¹ and R¹¹" groups when more than one of these groups is present. When R' and R" are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a A-, 5-, 6-, or 7-membered ring. For example, -NR¹R" is meant to include, but not be limited to, 1-pyrrolidinyl and 4-morpholinyl. From the above discussion of substituent moieties, one of skill in the art will understand that the term "alkyl" is meant to include groups including carbon atoms bound to groups other than hydrogen groups, such as haloalkyl (e.g., -CF₃ and -CH₂CF₃) and acyl (e.g., -C(O)CH₃, -C(O)CF₃, -C(O)CH₂OCH₃, and the like).

[0061] Similar to the substituent moieties described for the alkyl radical, substituent moieties for the aryl and heteroaryl groups are varied and may be selected from, for example: halogen, -OR', -NR¹R", -SR¹, -halogen, -SiR¹R¹¹R¹", -OC(O)R¹, -C(O)R¹, -CO₂R', - CONR¹R", -OC(O)NR¹R", -NR¹¹C(O)R', -NR'-C(O)NR"R"', -NR¹¹C(O)₂R¹, -NR-

C(NR¹R¹¹R¹O=NR"¹', -NR-C(NR¹R¹O=NR'", -S(O)R¹, -S(O)₂R¹, -S(O)₂NR¹R", -NRSO₂R¹, - CN and -NO₂, -R', -N₃, -CH(Ph)₂, UuOrO(C₁ -C₄)alkoxy, and fluoro(C₁-C₄)alkyl, in a number ranging from zero to the total number of open valences on the aromatic ring system; and where R, R", R'" and R"" are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl. When a compound of the invention includes more than one R group, for example, each of the R groups is independently selected as are each R, R", R'" and R"" groups when more than one of these groups is present.

[0062] Two of the substituent moieties on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -T-C(O)-(CRR')_q-U-, wherein T and U are independently -NR-, -O-, -CRR¹- or a single bond, and q is an integer of from 0 to 3. Alternatively, two of the substituent moieties on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -A-(CH₂)_r-B-, wherein A and B are independently -CRR¹-, -O-, -NR-, -S-, -S(O)-, -S(O)₂-, -S(O)₂NR'- or a single bond, and r is an integer of from 1 to 4. One of the single bonds of the new ring so formed may optionally be replaced with a double bond. Alternatively, two of the substituent moieties on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -(CRR')_s-X'~(C"R"')d-, where s and d are independently integers of from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O)₂-, Or-S(O)₂NR'-. The substituent moieties R, R, R" and R'" are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0063] As used herein, the term "heteroatom" or "ring heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0064] The term "pharmaceutically acceptable salts" is meant to include salts of the active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ,

ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable 5 acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, 0 suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al, "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and 5 acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0065] The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical 0 properties, such as solubility in polar solvents.

[0066] In addition to salt forms, the present invention provides compounds, which are in a prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Additionally, prodrugs can be converted to the compounds of the 5 present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0067] Certain compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent 0 to unsolvated forms and are encompassed within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0068] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; the racemates, diastereomers, tautomers, geometric isomers and individual isomers are encompassed within the scope of the present invention. The compounds of the present invention do not include those which are known in the art to be too unstable to synthesize and/or isolate.

[0069] The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). AU isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[0070] Where two groups are "optionally joined together to form a ring," the two groups are covalently bonded together with the atom or atoms to which the two groups are joined to form a substituted or unsubstituted aryl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted cycloalkyl, or a substituted or unsubstituted heterocycloalkyl ring.

[0071] The terms "arylalkyl," "heteroarylalkyl," "cycloalkyl-alkyl," and "heterocycloalkyl-alkyl," as used herein, refer to an aryl, heteroaryl, cycloalkyl and heterocycloalkyl, respectively, attached to the remainder of the molecule via an alkylene group. Where an "arylalkyl," "heteroarylalkyl," "cycloalkyl-alkyl," or "heterocycloalkyl- alkyl" is substituted, one or more substituent moieties may be covalently bonded to the alkylene moiety and/or the aryl, heteroaryl, cycloalkyl and heterocycloalkyl moieties, respectively. A "C₁-C₂₀" arylalkyl, heteroarylalkyl, cycloalkyl-alkyl, or heterocycloalkyl- alkyl, are moieties in which a C₁-C₂₀ alkylene links an aryl, heteroaryl, C₄-C₈ cycloalkyl, and 4 to 8 membered heterocycloalkyl, respectively, to the remainder of the molecule. A "C₁-Cs" arylalkyl, heteroarylalkyl, cycloalkyl-alkyl, or heterocycloalkyl-alkyl, are moieties in which a C₁-C₈ alkylene links an aryl, heteroaryl, C₅-C₇ cycloalkyl, and 5 to 7 membered heterocycloalkyl, respectively, to the remainder of the molecule

[0072] A "substituent group," as used herein, means a group selected from the following moieties: [0073] (A) -OH, -NH₂, -SH, -CN, -CF₃, oxy, halogen, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and

[0074] (B) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, substituted with at least one substituent selected from:

[0075] (i) oxy, -OH, -NH₂, -SH, -CN, -CF₃, halogen, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and

[0076] (ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, substituted with at least one substituent selected from:

[0077] ^> (a) oxy, -OH, -NH₂, -SH, -CN, -CF₃, halogen, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, and

[0078] (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, substituted with at least one substituent selected from oxy, -OH, -NH₂, -SH, -CN, -CF₃, halogen, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, and unsubstituted heteroaryl.

[0079] A "size-limited substituent" or " size-limited substituent group," as used herein means a group selected from all of the substituents described above for a "substituent group," wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C₁- C₂₀ alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C₄-C₈ cycloalkyl, and each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 4 to 8 membered heterocycloalkyl.

[0080] A "lower substituent" or " lower substituent group," as used herein means a group selected from all of the substituents described above for a "substituent group," wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C₁-C₈ alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8 membered 00183

heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C₅- C₇ cycloalkyl, and each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 5 to 7 membered heterocycloalkyl.

[0081] The term "treating" refers to any indicia of success in the treatment or amelioration of an injury, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving a patient's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subj ective parameters; including the results of a physical examination, neuropsychiatric exams, and/or a psychiatric evaluation. For example, the methods of the invention successfully treat a patient's delirium by decreasing the incidence of disturbances in consciousness or cognition.

[0082] The term "higher alkyl" refers to those alkyl groups having at least six carbon atoms. The term "lower alkyl" refers to those alkyl groups having from one to five carbon atoms.

DESCRIPTION OF THE EMBODIMENTS

I. COMPOUNDS OF THE PRESENT INVENTION

[0083] In one aspect, the present invention provides an antiproliferative, low-calcemic compound having the Formula (II) :

wherein L is selected from the following carbonyl side chains:

wherein n = I₅ 2;

m = 0-2;

P = 3-6;

Z = CH₂, O, NR', S₅ SO, SO₂, O-N-R', N(R')-N(R'), -(C=O)-;

Y = CCH₂;

X = C₅ O₅ S₅ N;

V = H₅ Or V = V = CH₂;

R = isoalkyl, t-butyl, l-(Methyl)cycloalkyl (3-7), l-(OH)cycloalkyl (3-7), 1-adamantyl;

R = H, F, Me, CD₃, Et, /-Pr₅ allyl, CH₂OR₅ CMe₂Ph, R=R=cycloalkyl (3-7); and

R" = H₅ F₅ Me₅ CD₃, Et₅ /-Pr, allyl, CH₂OR₅ R=R'=cycloalkyl (3-7).

[0084] In another embodiment, the compounds of the present invention has the formulas provided in Table 1. These compounds were prepared according to the synthesis schemes that follow in detail below.

Table 1

000183

TUS2006/000183

6000183

2006/000183

2006/000183

II. ASSAYS

Determination of Antiproliferative and Antitumor Activities

[0085] Growth Inhibition. To determine the inhibitory effect of the compositions of the present invention on cell proliferation, screening assays were performed on murine B16 malignant melanoma cells and growth curves for the tested compounds are ahson in figures 1-10.

[0086] Murine Bl 6 malignant melanoma cells were grown and propagated in RPMI medium supplemented with 10% fetal bovine serum, 1-glutamine, penicillin, and streptomycin and incubated at 37° C in 5% CO₂. For proliferation studies, cells were washed with PBS, trypsinized, and suspended in 8 mL of supplemented RPMI medium. The cell density was then determined using a hemacytometer, and cells were resuspended in RPMI at 10,000 cells/cm³. One milliliter of cell suspension (10,000 cells) was added to each of a Falcon 24 well flat bottom tissue culture plate (Becton-Dickinson, Lincoln Park, NJ). Plates were incubated for 24 h to allow for cell attachment. The medium was then removed and replaced with fresh RPMI medium containing either 0.4% solvent (2- propanol) or drug at concentrations ranging from 1 to 1000 nM in triplicate. When control wells neared confluence, cells were washed with PBS, trypsinized, and suspended in 10 mL of Isoton II Coulter balanced electrolyte solution in FISHER brand DiIu- Vial cuvettes. Cell number was then determined for each well as an average of two readings on a ZM Coulter counter. Results are expressed as the average cell number for each drug treatment group divided by the initial cell number (NZNo)-

[0087] The standard in vitro murine keratinocyte assay (Posner, G. H., et al, J. Med. Chem., 41:3008-3014 (1998)) described briefly above indicates that the 25-oxo analogs Ia, and 4a, as well as the 24-oxo analogs 3 and 5 are much more antiproliferative than natural calcitriol (I) as is especially notable at physiologically relevant low nanomolar concentrations (Fig. 1).

[0088] Noteworthy is that analog 3, differing only slightly from natural calcitriol (I) by being a 24-ketone and by having a methyl group in place of the classical 25-OH group, is desirably much more antiproliferative than calcitriol (I). Noteworthy also for SAR generalizations is that there is little observed difference in antiproliferative activity between the pair of saturated versus α,/3-unsaturated ketones Ia versus 4b. 24,24-Difluorinated analog 6 and 24,24-dimethylated analog 7, however, are much less antiproliferative than calcitriol (I) and than the corresponding 24-unsubstituted analog Ia (data not shown); thus, both electron- withdrawing groups (i.e., F) and a sterically hindered 25-carbonyl as in analog 7 dimmish antiproliferative activity considerably.

Determination of Urinary Calcium Levels

[0089] Male F344 rats (150 g) were housed individually in glass metabolism cages and received food and water ad libitum. After several days acclimation, rats received 1 μg/kg of body weight of test glycol/0.05 M Na₂HPO₄ (80:20). Urine samples, which were collected on ice, were centrifuged at 650g for 10 min, adjusted to pH 6.0 as necessary, and assayed for calcium content spectrophotometrically at 575 nm using reagents and standards from Sigma calcium kit no. 587. The effectsof the compounds of the present invention on urinary excretion in rats can be seen in figures 11-15. [0090] Our standard in vivo mouse urine assay (Posner, G. H., et al, J. Med. Chem.,

41:3008-3014 (1998)) described briefly above shows that at least 10 times higher doses of the saturated ketone analogs Ia and 3 are required to cause the same level of urinary calcium excretion as the natural hormone calcitriol (I, Fig. 11). [0091] When dosed at the same level as calcitriol (I), ketone Ia was much less calcemic than calcitriol (I). Introduction of a conjugated C=C double bond as in enone 4a and 4b does not alter calcemic activity very much (Fig. 11). Surprisingly, based on DeLuca's observations that 19-nor analogs usually are much less calcemic than their 19-methylene versions, Uskokovic, M. R., et al, P. Curr. Pharm. Des., 3:99-123 (1997) and Sicinski, R. R., et al, J. Med. Chem., 45:3366 (2002), the 19-nor version of 25-ketone Ia (i.e., 19-nor- Ia or compound 2) does not have significantly less calcemic activity than its 19-methylene counterpart Ia. _f

[0092] Two unexpected observations arise from this work: (1) simply replacing the natural 25-OH group in calcitriol (I) by a-CH3 group and incorporating a 24-ketone functionality produces analog 3 that is substantially more antiproliferative in vitro than the natural hormone even at physiologically relevant nanomolar concentrations; and (2) simply 5 removing the one carbon exocyclic 19- methylene group produces analog 2 (i.e., 19-nor-la) that is not significantly less calcemic than 19-methylene analog Ia. These important SAR generalizations may help design even more potent and safe new analogs of calcitriol (I).

[0093] In another aspect, the present invention provides pharmaceutical compositions. The pharmaceutical composition includes a pharmaceutically acceptable excipient and a 10 compound having the formula (II) :

wherein L is selected from the following carbonyl side chains:

wherein n = 1, 2;

m = 0-2;

p = 3-6;

Z = CH₂, O, NR¹, S₅ SO₅ SO₂, 0-N-R, N(RO-N(R¹), -(C=O)-;

20 Y = O₅CH₂;

X = C₅ O₅ S₅ N; _t

V = H₅ OrV = V = CH₂;

R = isoalkyl, f-butyl, l-(Methyl)cycloalkyl (3-7), l-(OH)cycloalkyl (3-7), 1-adamantyl;

R' = H, F, Me, CD₃, Et, /-Pr, allyl, CH₂OR', CMe₂Ph, R'=R'=cycloalkyl (3-7); and

R" = H, F, Me, CD₃, Et, z-Pr, allyl, CH₂OR', R'=R'=cycloalkyl (3-7).

5 [0094] The pharmaceutical compositions described herein are typically used for chemotherapy of various human illnesses, such as but not limited to osteoporesis, psoriasis, kidney failure, and immunosuppressant disorders in a subject in need of such treatment.

[0095] In an exemplary embodiment, the pharmaceutical composition includes from 1 to 2000 milligrams of the compound of Formula (II). hi some embodiments, the 0 pharmaceutical composition includes from 1 to 1500 milligrams of the compound of Formula (II). In other embodiments, the pharmaceutical composition includes from 1 to 1000 milligrams of the compound of Formula (II).

[0096] The compounds of the present invention can be prepared and administered in a wide variety of oral, parenteral and topical dosage forms. Oral preparations include tablets,

15 pills, powder, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions, etc., suitable for ingestion by the patient. The compounds of the present invention can also be administered by injection, that is, intravenously, intramuscularly, intracutaneously, subcutaneously, intraduodenally, or intraperitoneally. Also, the compounds described herein can be administered by inhalation, for example, intranasally. Additionally, the 0 compounds of the present invention can be administered transdermally. The compounds of this invention can also be administered by in intraocular, intravaginal, and intrarectal routes including suppositories, insufflation, powders and aerosol formulations (for examples of steroid inhalants, see Rohatagi, J. Clin. Pharmacol. 35:1187-1193, 1995; Υyw&, Ann. Allergy Asthma Immunol. 75:107-111, 1995). Thus, the pharmaceutical compositions

25 described herein may be adapted for oral administration. In some embodiments, the pharmaceutical composition is in the form of a tablet. Moreover, the present invention provides pharmaceutical compositions including a pharmaceutically acceptable carrier or excipient and either a compound of Formula (II), or a pharmaceutically acceptable salt of a compound of Formula (II).

30 [0097] For preparing pharmaceutical compositions from the compounds of the present invention, pharmaceutically acceptable carriers can be either solid or liquid. Solid form „_,._,

preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances, which may also act as diluents, flavoring agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. Details on techniques for formulation and administration are well described in the 5 scientific and patent literature, see, e.g., the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's").

[0098] hi powders, the carrier is a finely divided solid, which is in a mixture with the finely divided active component, hi tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape 10 and size desired.

[0099] The powders and tablets preferably contain from 5% or 10% to 70% of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term 15 "preparation" is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is surrounded by a carrier, which is thus in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

20 [0100] Suitable solid excipients are carbohydrate or protein fillers include, but are not limited to sugars, including lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato, or other plants; cellulose such as methyl cellulose, hydroxypropylmethyl- cellulose, or sodium carboxymethylcellulose; and gums including arabic and tragacanth; as well as proteins such as gelatin and collagen. If desired, disintegrating or solubilizing

25 agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate.

[0101] Dragee cores are provided with suitable coatings such as concentrated sugar solutions, which may also contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic 30 solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for product identification or to characterize the quantity of active compound (i.e., dosage). Pharmaceutical preparations of the invention can also be used orally using, for example, push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a coating such as glycerol or sorbitol. Push-fit capsules can contain the compounds of this invention mixed with a filler or binders such as lactose or starches, lubricants such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the compounds of this invention compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycol with or without stabilizers.

[0102] For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active component is dispersed homogeneously therein, as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool, and thereby to solidify.

[0103] Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water/propylene glycol solutions. For parenteral injection, liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.

[0104] Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethylene oxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or a condensation product of ethylene oxide with a partial ester derived from fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate). The aqueous suspension can also contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, aspartame or saccharin. Formulations can be adjusted for osmolality.

[0105] Also included are solid form preparations, which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

[0106] Oil suspensions can be formulated by suspending a compound of this invention in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin; or a mixture of these. The oil suspensions can contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents can be added to provide a palatable oral preparation, such as glycerol, sorbitol or sucrose. These formulations can be preserved by the addition of an antioxidant such as ascorbic acid. As an example of an injectable oil vehicle, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997. The pharmaceutical formulations of the invention can also be in the form of oil-in- water emulsions. The oily phase can be a vegetable oil or a mineral oil, described above, or a mixture of these. Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan mono-oleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan mono-oleate. The emulsion can also contain sweetening agents and flavoring agents, as in the formulation of syrups and elixirs. Such formulations can also contain a demulcent, a preservative, or a coloring agent.

[0107] The compounds of this invention of the invention can be delivered by transdermally, by a topical route, formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.

[0108] The compounds of this invention of the invention can also be delivered as microspheres for slow release in the body. For example, microspheres can be administered via intradermal injection of drug-containing microspheres, which slowly release subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645, 1995; as biodegradable and injectable gel formulations (see, e.g., Gao Pharm. Res. 12:857-863, 1995); or, as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674, 1997) . Both transdermal and intradermal routes afford constant delivery for weeks or months.

[0109] The compounds of this invention pharmaceutical formulations of the invention can be provided as a salt and can be formed with many acids, including but not limited to 2006/000183

hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend to be more soluble in aqueous or other protonic solvents that are the corresponding free base forms. In other cases, the preparation may be a lyophilized powder in 1 mM-50 mM histidine, 0.1%- 2% sucrose, 2%-7% marmitol at a pH range of 4.5 to 5.5, that is combined with buffer prior to use.

[0110] In another embodiment, the compounds of this invention formulations of the invention are useful for parenteral administration, such as intravenous (IV) administration or administration into a body cavity or lumen of an organ. The formulations for administration will commonly comprise a solution of the compounds of this invention dissolved in a pharmaceutically acceptable carrier. Among the acceptable vehicles and solvents that can be employed are water and Ringer's solution, an isotonic sodium chloride. In addition, sterile fixed oils can conventionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can likewise be used in the preparation of injectables. These solutions are sterile and generally free of undesirable matter. These formulations may be sterilized by conventional, well known sterilization techniques. The formulations may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents, e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of the compounds of this invention in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight, and the like, in accordance with the particular mode of administration selected and the patient's needs. For IV administration, the formulation can be a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally-acceptable diluent or solvent, such as a solution of 1,3-butanediol.

[0111] In another embodiment, formulations having the compounds of this invention can be delivered by the use of liposomes which fuse with the cellular membrane or are endocytosed, i.e., by employing ligands attached to the liposome, or attached directly to the oligonucleotide, that bind to surface membrane protein receptors of the cell resulting in endocytosis. By using liposomes, particularly where the liposome surface carries ligands _t

specific for target cells, or are otherwise preferentially directed to a specific organ, one can focus the delivery of the compounds of this invention into the target cells in vivo. (See, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, ^m. J. Hosp. Pharm. 46:1576-1587, 1989).

5 [0112] The pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it 10 can be the appropriate number of any of these in packaged form.

[0113] The quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 10000 mg, more typically 1.0 mg to 1000 mg, most typically 10 mg to 500 mg, according to the particular application and the potency of the active component. The composition can, if desired, also contain other compatible therapeutic agents.

15 II. METHODS FOR TREATING CONDITIONS IN NEED OF CHEMOTHERAPY

[0114] hi still another aspect, the present invention provides a method for the treatment of a disorder or condition which requires chemotherapy, m this method, a subject in need of such treatment is administered an effective amount of a compound having one of the 20 formulae provided above. The amount is effective in treating the illness that the patient is afflicted with.

[0115] A variety of disease sates are capable of being treated with chemotherapy. Exemplary disease states include, but are not limited to cancer, kidney failure, osteoporesis, psoriasis, kidney failure, and immunosuppressant disorders. The methods of treatment 25 includes administering to a patient in need of such treatment, a therapeutically effective amount of a compound according to Formula (II), or a pharmaceutically acceptable salt thereof.

[0116] Thus, in an exemplary embodiment, the present invention provides a method of treating a disorder or condition through chemotherapy, the method including administering 30 to a subject in need of such treatment, an effective amount of a compound of the present invention, such as a compound of Formula (II). j

[0117] The amount of the compounds of this invention adequate to treat a disease is defined as a "therapeutically effective dose". The dosage schedule and amounts effective for this use, i.e., the "dosing regimen," will depend upon a variety of factors, including the stage of the disease or condition, the severity of the disease or condition, the general state of 5 the patient's health, the patient's physical status, age and the like. In calculating the dosage regimen for a patient, the mode of administration also is taken into consideration.

[0118] The dosage regimen also takes into consideration pharmacokinetics parameters well known in the art, i.e., the rate of absorption, bioavailability, metabolism, clearance, and the like (see, e.g., Hidalgo-Aragones (1996) J. Steroid Biochem. MoI. Biol. 58:611-617; 10 Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sd. 84:1144-1146; Rohatagi (1995) Pharmazie 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol. 24:103-108; the latest Remington's, supra). The state of the art allows the clinician to determine the dosage regimen for each individual patient, and disease or condition treated.

15 [0119] Single or multiple administrations of formulations having the compounds of this invention can be administered depending on the dosage and frequency as required and tolerated by the patient. The formulations should provide a sufficient quantity of active agent to effectively treat the disease state. Thus, in one embodiment, the pharmaceutical formulations for oral administration of the compounds of this invention is in a daily amount

20 of between about 0.5 to about 20 mg per kilogram of body weight per day. In an alternative embodiment, dosages are from about 1 mg to about 4 mg per kg of body weight per patient per day are used. Lower dosages can be used, particularly when the drug is administered to an anatomically secluded site, such as the cerebral spinal fluid (CSF) space, in contrast to administration orally, into the blood stream, into a body cavity or into a lumen of an organ.

25 Substantially higher dosages can be used in topical administration. Actual methods for preparing parenterally administrable formulations having the compounds of this invention will be known or apparent to those skilled in the art and are described in more detail in such publications as Remington's, supra. See also Nieman, In "Receptor Mediated Antisteroid Action," Agarwal, et al., eds., De Grayter, New York (1987).

30 [0120] After a pharmaceutical composition including a compound of the invention has been formulated in an acceptable carrier, it can be placed in an appropriate container and labeled for treatment of an indicated condition. For administration of the compounds of this ,

invention, such labeling would include, e.g., instructions concerning the amount, frequency and method of administration. In one embodiment, the invention provides for a kit for the treatment of cancer or kidney failure in a human which includes the compounds of this invention and instructional material teaching the indications, dosage and schedule of 5 administration of the compounds of this invention.

III. EXEMPLARY SYNTHESES

[0121] The compounds of the invention are synthesized by an appropriate combination of generally well known synthetic methods. Techniques useful in synthesizing the compounds of the invention are both readily apparent and accessible to those of skill in the relevant art. 10 The discussion below is offered to illustrate certain of the diverse methods available for use in assembling the compounds of the invention. However, the discussion is not intended to define the scope of reactions or reaction sequences that are useful in preparing the compounds of the present invention.

IV. EXAMPLES

15 [0122] As can be appreciated by the skilled artisan, methods of synthesizing the compounds of the described herein will be evident to those of ordinary skill in the art. Additionally, the various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing the compounds

20 described herein are known in the art and include, for example, those such as described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d. Ed. , John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic

25 Synthesis, John Wiley and Sons (1995), and subsequent editions thereof.

[0123] The discussion below is offered to illustrate certain of the diverse methods available for use in assembling the compounds of the invention. However, the discussion is not intended to define the scope of reactions or reaction sequences that are useful in preparing the compounds of the present invention. Experimental Section:

[0124] Unless otherwise noted, reactions were run in flame-dried round-bottomed flasks under an atmosphere of ultra-high-purity (UHP) argon. AU reactive liquid reagents were transferred by syringe or cannula and were added into the flask through a rubber septum. Tetrahydrofuran was freshly distilled from sodium benzophenone ketyl immediately prior to use. All other solvents and reagents were used as received unless otherwise stated, n- BuLi was obtained from commercial sources and was titrated with JV-pivaloyl-O-toluidine prior to use. Diethyl ether (ether) and tetrahydrofuran (THF) were distilled from sodium benzophenone ketyl prior to use. Methylene chloride (CH₂Cl₂) was distilled from calcium hydride prior to use. AU other compounds were purchased from Aldrich Chemical Co. and used without further purification. Analytical thin-layer chromatography (TLC) was conducted with silica gel 60 F254 plates (250 Im thickness; Merck). Column chromatography was performed using short path silica gel (particle size <230 mesh), flash silica gel (particle size 400-230 mesh), or Florisil (200 mesh). Yields are not optimized. Purity of products was judged to be >95% based on their chromatographic homogeneity. High-performance liquid chromatography (HPLC) was carried out with a Rainin HPLX system equipped with two 25 mL/min preparative pump heads using Rainin Dynamax 10 250 mm (semipreparative) columns packed with 60 A ° silica gel (8 Im pore size), either as bare silica or as C-18-bonded silica. Melting points were measured using a Mel-Temp metal-block apparatus and were uncorrected. Nuclear magnetic resonance (NMR) spectra were obtained either on a Varian XL-400 spectrometer, operating at 400 MHz for ¹H and 100 MHz for ¹³C, or on a Varian XL-500 spectrometer, operating at 500 MHz for ¹H and 125 MHz for ¹³C. Chemical shifts are reported in parts per million (ppm, δ) downfield from tetramethylsilane (TMS). Multiplicities of signals in the ¹H NMR spectra are reported as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd (doublet of doublets), dt (doublet of triplet), etc.

[0125] Infrared spectra were obtained on a Perkin Elmer 1600 FT-IR spectrometer as liquid films and thin layer with NaCl cells. Intensities were reported as s (strong 67- 100%), m (medium 34-66%), and w (weak 0-33%) with the following notations, br (broadened), sh (shoulder), etc. ₍

[0126] Optical rotations were recorded on JASCO, P-1100 model polarimeter (Japan Spectroscopic Co., Ltd.) with sodium D line at the temperatures as indicated in the experimental section for the specific compounds.

[0127] Analytical Thin Layer Chromatography (TLC) was performed on Merck silica 5 gel plates (Merck Kieselgel, 60, 0.25-mm thickness) with F₂₅₄ indicator. Compounds were visualized under UV lamp and/or by developing with iodine, vanillin, p-anisaldehyde or KMnO₄ followed by heating with a heat gun. Flash chromatography (Still, W. C, et al., J. Org. Chem., 43:1404 (1978)) was performed as reported by Still and coworkers on 230- 400 mesh silica gel (E.M. Science) with technical and/or FIPLC grade solvents. Medium

10 Pressure Liquid Chromatography (MPLC) was performed with FMI pump and prepacked silica gel column (Merck, Labor Columns, LiChroprep Si 60, 40-63 μm). High Pressure Liquid Chromatography (HPLC) was performed on a Rainin HPLX system equipped with two 25 mL pump heads and a Rainin Dynamax UV-C dual-beam variable wavelength detector set at 254 or 260 nm using Phenomenex, Luna 5 μ C18 semipreparative (250 x 10

15 mm) column and Chiralcel OJ semipreparative (250 x 10 mm) column.

[0128] Low and high-resolution mass spectra (LRMS and HRMS) were obtained with electronic or chemical ionization (EI or CI) either (1) at Johns Hopkins University on a VG Instruments 70-S spectrometer run at 70 eV for EI and run with ammonia (NH₃) as a carrier gas for CI or (2) at the Ohio State University on a Finnigan- MAT CH5, a 20 Finnigan-MAT 731 , or a VG Instruments 70-VSE spectrometer run at 70 eV for EI and run with methane (CH₄) for CL

[0129] The t-butyl-iV-(H), t-butyl-iV-(Me) hydroxamic acids, iodide (+)-25, and Tosylate 83 were prepared according to literature precedent (Barbaric, M., et al, J. Med. Chem., 48:884-887 (2005); Posner, G. H., et al, J. Med. Chem., 41:3008 (1998); Posner, G.H., et 25 al, J. Med. Chem., 42:3425 (1999); and Craig, D., et al, Tetrahedron, 55:15025 (1999)).

[0130] Synthesis of new analogs Ia, 19-nor-la (compound 2), and 3 is described in Examples 1-3 and outlined schematically in Figures 10-13, starting from previously reported side-chain iodides 22 (Posner, G. H., et al, J. Med. Chem., 41,:3008-3014 (1998)) and 25 (Dai, H., et al, Synthesis, 1383-1398 (1994)). As noted previously in the 16-ene 30 series (Posner, G. H., et al, J. Med. Chem., 41,:3008-3014 (1998)), so now in the 16,17- saturated series, Horner-Wadsworth-Emmons (HWE) coupling of the A-ring nucleophile with 8,24- and 8,25-diketones 29 and 30 proceeds regiospecifically at only C-8, the less 83

hindered detone carbonyl group. Synthesis of new enone analog 4a and 4b is summarized in Example 4, Figure 13, featuring successful HWE coupling of triene aldehyde 34 without compromising the essential but sensitive conjugated triene unit. Synthesis of new analog 5 is outlined in Example 5, Figure 14 and synthesis of analogs 6 and 7 is described in Examples 6 and 7, respectively and shown in Figure 15.

EXAMPLE l 25(0) Analogs (+VIa and (+VIb (Figure 16)

[0131] A. Preparation of tert-Butyl C,D-ring ketone 30. A 10-mL round-bottomed flask was charged with diisopropylamine (322 μL, 2.29 mmol distilled over calcium hydride prior to use) and 2 mL of distilled THF. This solution was cooled to -78 ⁰C, and n-BuLi (1.39 mL of 1.6 M solution in hexane, 2.23 mmol) was added via syringe. Pinacolone (272 JU.L, 2.17 mmol dried over potassium carbonate and activated molecular sieve for 24 hours immediately prior to use) was dissolved in 1 mL of distilled THF and cooled to -78 ⁰C at which point it was added to the reaction flask via cannula. The reaction was left to stir for 30 minutes. Hexamethylphosphoramide (HMPA, 300 μL) was then added via syringe, and the reaction mixture was allowed to stir for an additional 15 minutes. A solution of iodide 22 (140 mg, 0.31 mmol) in 2 mL of THF was cooled to -78 °C and added to the reaction mixture via cannula. The reaction mixture was stirred at -78 ⁰C for 2 hours and allowed to warm to room temperature slowly. The resulting yellow mixture was quenched with 2 mL of water, extracted with ethyl acetate (3x 25 mL), dried over MgSO₄, concentrated, and purified using silica gel column chromatography (4% ethyl acetate/hexanes) to give 24 as a colorless oil (130 mg, 97%). A 15-mL round-bottomed flask was charged with TES tert- butyl ketone 24 (130 mg, 0.30 mmol) dissolved in 5 mL of distilled THF.

Tetrabutylammonium fluoride (TBAF, 1 M solution in THF, 369 μL, 0.36 mmol) was added to the reaction flask, and this solution was left to stir at room temperature for 7 hours. The resulting mixture quenched with 2 mL of water, extracted with ethyl acetate (3x 25 mL), dried over MgSO₄, concentrated, and purified using silica gel column chromatography (10% ethyl acetate/hexanes) to give a colorless oil (80 mg, 84%): [α]" +20.8 (c 0.65, CHCl₃); ¹HNMR (400 MHz, CDCl₃) δ 4.04 (m, IH), 2.42-2.39 (m, 2H), 1.97-1.94 (m, IH), 1.84-1.76 (m, 3H), 1.59-1.15 (m, 8H), 1.10 (s, 9H), 0.90 (s, 3H), 0.89 (d, J= 7.6 Hz, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.1, 66.3, 56.3, 52.5, 44.0, 41.8, 40.3, 36.8, 35.3, 35.1, 33.5, 27.1, 26.3, 22.4, 20.2, 18.4, 17.3, 13.5; IR (neat, cm ⁴) 3358, 2932, 2872, 1701, 1365, 1152, 1023; HRMS m/z [M + Na⁺] calcd 331.2607 for C₂₀H₃₆O₂Na⁺, found 331.2590.

[0132] A flame-dried 25-mL flask equipped with a magnetic stir bar, and a septum along with an Ar ballon was charged with the alcohol tert-butyl ketone (80 mg, 0.25 mmol) and 5 dissolved in 2 niL freshly distilled CH₂Cl₂. Then, to this solution, PDC (273 mg, 0.72 mmol) was added. The resulting mixture was allowed to stir at room temperature for 8 hours. The mixture were filtered with Celite, concentrated, and purified using silica gel chromatography (10% ethyl acetate/hexanes) to give diketone 30 as a colorless oil (79 mg,

99%): [ocg +7.6 (c 0.75, CHCl₃). ¹H NMR (400 MHz, CDCl₃) δ 2.44 (m, 3H), 2.29-2.16 10 (m, 2H), 2.13-2.08 (m, IH), 2.03-1.80 (m, 2H), 1.77-1.25 (m, 12H), 1.12 (s, 9H), 0.97 (d, J = 6.4 Hz, 3H), 0.62 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.0, 212.1, 62.0, 56.3, 49.9, 44.1, 40.9, 38.9, 36.6, 35.4, 35.3, 27.4, 26.4, 24.0, 20.1, 19.0, 18.6, 12.4; IR (neat, cm^"1) 2958, 2873, 1711, 1478, 1367, 1073; HRMS m/z [M + Na⁺] calcd 329.2451 for C₂₀H₃₄O₂Na⁺, found 329.2469.

15 [0133] B. Preparation of 25(O) Analogs (+)-l a and (+)-lb. Racemic phosphine oxide (±)-31 and enantiomerically pure C,D-ring ketone (+)-30, each was separately azeotropically dried with anhydrous benzene (4x 1 mL) on a rotary evaporator and held under vacuum (ca. 0.1 mmHg) for at least 48 hours prior to use. A flame-dried 10-mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon was

20 charged with racemic phosphine oxide (±)-31 (70 mg, 0.12 mmol) which was dissolved in 2 mL freshly distilled THF. The flask was cooled down to -78 ⁰C in a dry ice bath. To this solution, n-BuLi (82 μL, 0.12 mmol, 1.6 M solution in hexanes) was added dropwise over several minutes, during which time a deep red color was developed and persisted. This mixture was allowed to stir at -78 ⁰C for an additional 10 minutes. Meanwhile, a fiame-

25 dried 10-mL recovery flask equipped with a magnetic stir bar, and a septum along with an Ar balloon was charged with C,D-ring ketone 30 (33 mg, 0.1 mmol), dissolved in 1 mL freshly distilled THF, and cooled down to -78 ⁰C in a dry ice bath. The solution of C,D-ring ketone was gently transferred into the flask containing the phosphine oxide anion at -78 ⁰C over several minutes. After the addition was complete, the deep red color persisted and the

30 mixture was allowed to stir at -78 ⁰C for 8 hours, during that time it was visually checked. On observation of the light yellow color, the reaction was quenched at -78 ⁰C by adding 3 mL of pH 7 buffer and allowed to come to room temperature. The mixture was then rinsed into a separatory funnel with ethyl acetate and extracted with ethyl acetate (3x 25 mL). The p T/US2006/000183

combined extracts were washed with water (Ix 25 niL) and brine solution, dried over MgSO₄, filtered, and purified using silica gel chromatography (5% ethyl acetate/ hexanes) to give a colorless oil (40 mg, 55%). A 15-mL round-bottomed flask was charged with the coupled compound (20 mg, 0.02 mmol) dissolved in 5 mL of distilled THF. 5 Tetrabutylammonium fluoride (TBAF, 1 M solution in THF, 148 μL, 0.1 mmol) was added to the reaction flask, and this solution was left to stir at room temperature for 7 hours. The resulting mixture was quenched with 2 mL of water, extracted with ethyl acetate (3x 25 mL), dried over MgSO₄, concentrated, and purified using silica gel column chromatography (25% ethyl acetate/hexanes) to give Ia and Ib (11 mg, 83%) as a mixture of diastereomers. 10 The diastereomeric mixture was separated by an HPLC using Chiral OD [semipreparative (Ix 25 cm), flow rate = 2.5 mL/min] eluted with 2.5% isopropyl alcohol in hexanes to afford 4 mg of Ia (lα , 3β) and 1.3 mg of Ib (IB, 3α) in 55 and 17% yield. The retention time for Ia is 103.15 min and Ib is 121.28 minutes. Data for Ia (lα, 3B): [αg +20.3 (c 0.1, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.36 (d, J= 11.2 Hz, IH), 5.99 (d, J= 11.2 Hz,

15 IH), 5.31 (s, IH), 4.98 (s, IH), 4.41 (m, IH), 4.21 (m, IH), 2.80 (dd, J= 12.0, 4.0 Hz, IH), 2.58 (dd, J= 12.0, 2.8 Hz, IH), 2.42 (m, 2H), 2.29 (dd, J= 13.6, 6.4 Hz, IH), 2.03-1.82 (m, 5H), 1.66-1.62 (m, 2H), 1.53-1.51 (m, 3H), 1.48-1.23 (m, 10H), 1.11 (s, 9H), 0.92 (d, J= 6.4 Hz, 3H), 0.515 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.2, 147.6, 143.3, 132.8, 125.0, 116.9, 111.8, 70.8, 66.9, 56.3, 56.2, 45.9, 45.3, 44.1, 42.8, 40.4, 36.9, 36.0, 35.5,

20 29.1, 27.6, 26.4, 23.6, 22.2, 20.4, 18.8, 12.0; IR (neat, cm^"1) 3374, 2949, 2360, 1705, 1464, 1366, 1054; HRMS m/z [M + Na⁺] calcd 465.3339 for C₂₉H₄₆O₃Na⁺, found 465.3310. Data for Ib (IB, 3α): [aj? +3.9 (c 0.1, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.39 (d, J= 11.2 Hz, IH), 6.05 (d, J= 11.2 Hz, IH), 5.32 (s, IH), 5.00 (s, IH), 4.44 (m, IH), 4.21 (m, IH), 2.82 (dd, J- 12.0, 3.6 Hz, IH), 2.61 (dd, J= 13.2, 4.0 Hz, IH), 2.44 (m, 3H), 2.30 (dd, J=

25 13.2, 7.6 Hz, IH), 2.04-1.82 (m, 5H), 1.68-1.63 (m, 2H), 1.55-1.49 (m, 3H), 1.47-1.25 (m, 10H), 1.31 (s, 9H), 0.94 (d, J= 6.4 Hz, 3H), 0.54 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.2, 247.2, 143.3, 132.6, 125.0, 117.0, 112.6, 94.4, 71.4, 66.8, 56.3, 56.2, 45.9, 45.5, 42.8, 40.4, 36.9, 36.0, 35.5, 29.0, 27.6, 26.4, 23.6, 22.3, 20.4, 18.8, 12.0; IR (neat, cm^"1) 3368, 2951, 1705, 1465, 1054; HRMS m/z [M + Na⁺] calcd 465.3339 for C₂₉H₄₆O₃Na⁺, found

30 465.3332. EXAMPLE 2

19-nor-25(O) Analog (+)-2 (Figure 17)

[0134] Preparation of 19-nor-25(0) Analog (+)-2. 19-nor Phosphine oxide (+)-31a and enantiomerically pure C,D-ring ketone (+)-30, each was separately azeotropically dried with anhydrous benzene. (4x 1 niL) on a rotary evaporator and held under vacuum (ca. 0.1 lnmHg) for at least 48 hours prior to use. A flame-dried 10-mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon was charged with phosphine oxide (+)-31 (60 mg, 0.12 mmol) which was dissolved in 2 niL freshly distilled THF. The flask was cooled down to -78 ⁰C in a dry ice bath. To this solution was added n-BuLi (81 μL, 0.12 mmol, 1.42 M solution in hexanes) dropwise over several minutes, during which time a deep red color was developed and persisted. This mixture was allowed to stir at -78 ⁰C for an additional 10 minutes. Meanwhile, a flame-dried 10-mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon was charged with C,D-ring ketone (+)- 30 (27 mg, 0.08 mmol), dissolved in 1 mL freshly distilled THF, and cooled down to -78 ⁰C in a dry ice bath. The solution of C,D-ring ketone was gently transferred into the flask containing the phosphine oxide anion at -78 ⁰C via cannula over several minutes. After the addition was complete, the deep red color persisted and the mixture was allowed to stir at -78 ⁰C for 8 hours, during which time it was visually checked. On observation of the light yellow color, the reaction was quenched at -78 ⁰C by adding 3 mL of pH 7 buffer and allowed to come to room temperature. The mixture was then rinsed into a separatory funnel with ethyl acetate and extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL) and brine solution, dried over MgSO₄, filtered, and purified using silica gel chromatography (2% ethyl acetate/hexanes) to give a colorless oil (8 mg, 15%). A 15-mL round-bottomed flask was charged with the coupled compound (8 mg, 0.02 mmol) dissolved in 5 mL of distilled THF. Tetrabutylammonium fluoride (TBAF, 1 M solution in THF, 148 μL, 0.1 mmol) was added to the reaction flask, and this solution was left to stir at room temperature for 7 hours. The resulting mixture was quenched with 2 mL of water, extracted with ethyl acetate (3x 25 mL), dried over MgSO₄, concentrated, and purified using silica gel column chromatography

(25% ethyl acetate/hexanes) to give 19-nor-l (compound 2) (4 mg, 85%): [α£^s +15.2 (c 0.5, CHCl₃); ¹HNMR (400 MHz, CDCl₃) δ 6.31 (d, J= 11.6 Hz, IH), 5.85 (d, J= 11.2 Hz, IH), 4.41 (m, IH), 4.05 (m, IH), 2.81-2.71 (m, 2H), 2.50- 2.18 (m, 5H), 2.11-1.22 (m, 18H), j

1.13 (s, 9H), 0.94 (d, J= 6.8 Hz, 3H), 0.53 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.2, 143.2, 131.0, 123.9, 115.2, 67.4, 67.2, 56.3, 56.2, 45.8, 44.7, 42.2, 40.4, 37.2, 36.9, 36.0, 35.5, 29.7, 28.9, 27.6, 26.4, 23.5, 22.2, 20.4, 18.8, 12.0; IR (neat, cm ^A) 3500, 2926, 1706, 1364, 1049; HRMS m/z [M + Na⁺] calcd 453.3339 for C₂₈H₄₆O₃Na⁺, found 453.3351; UV 5 (CH₂Cl₂) )w = 255 nm (e = 29,530).

EXAMPLE 3 24(O)TB analog (+V3 (Figure 18)

10 [0135] A. Preparation of tert-Butyl C,D-ring ketone 29. A 10-mL round-bottomed flask was charged with diisopropylamine (207 μL, 1.41 mmol distilled over calcium hydride prior to use) and 2 mL of distilled THF. This solution was cooled to -78 ⁰C, and n-BuLi (1.0 ml of 1.6 M solution in hexane, 1.37 mmol) was added via syringe. Pinacolone (174 μL, 1.33 mmol dried over potassium carbonate and activated molecular sieve for 24 hours

15 immediately prior to use) was dissolved in 1 mL of distilled THF and cooled to -78 ⁰C at which point it was added to the reaction flask via cannula. The reaction was left to stir for 30 minutes. Hexamethylphosphoramide (HMPA, 300 μL) was then added via syringe, and the reaction mixture was allowed to stir for an additional 15 minutes. A solution of iodide 25 (0.09 g, 0.19 mmol) in 2 mL of THF was cooled to -78 °C and added to the reaction

20 mixture via cannula. The reaction mixture was stirred at -78 °C for 2 hours and allowed to warm to room temperature slowly. The resulting yellow mixture was quenched with 2 mL of water, extracted with ethyl acetate (3x 25 mL), dried over MgSO₄, concentrated, and purified using silica gel column chromatography (10% ethyl acetate/hexanes) to give a colorless oil 27 (0.06 g, 80%). A 15-mL round-bottomed flask was charged with TES-tert-

25 butyl ketone 27 (0.06 g, 0.2 mmol) dissolved in 5 mL of distilled THF.

Tetrabutylammonium fluoride (TBAF 1 M solution in THF, 440 μL, 0.4 mmol) was added to the reaction flask, and this solution was left to stir at room temperature for 7 hours. The resulting mixture was quenched with 2 mL of water, extracted with ethyl acetate (3x 25 mL), dried over MgSO₄, concentrated, and purified using silica gel column chromatography

30 (20% ethyl acetate/hexanes) to give a colorless oil (34 mg, 80%): [a]* +20.4 (c 2.0, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 4.05 (m, IH), 2.51-2.35 (m, 2H), 1.98-1.94 (m, IH), 1.89-1.68 (m, 4H), 1.56-1.29 (m, HH), 1.12 (s, 9H), 0.91 (s, 3H), 0.86 (d, J= 6.4 Hz, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.5, 69.3, 56.5, 52.5, 44.2, 41.8, 40.3, 34.9, 33.5, US2006/000183

33.3, 29.9, 27.0, 26.4, 22.5, 18.4, 17.4, 13.5; IR (neat, cm^'1) 3516, 2935, 2870, 1703, 1477, 1365, 1067, 990; HRMS m/z [M + Na⁺] calcd 317.2450 for C₁₉H₃₄O₂Na⁺, found 317.2436. A flame-dried 25 mL flask equipped with a magnetic stir bar, and a septum along with argon (Ar) ballon was charged with the alcohol tert-butyl ketone (0.035 g, 0.11 mmol) and dissolved in 2 mL freshly distilled CH₂Cl₂. Then, to this solution, PDC (125 mg, 0.32 mmol) was added. The resulting mixture was allowed to stir at room temperature for 8 hours. The mixture were filtered with Celite, concentrated, and purified using silica gel chromatography (30% ethyl acetate/hexanes) to give diketone 29 as a colorless oil (31 mg,

90%): [αg +11.5 (c 0.01, CHCl₃); ¹R NMR (400 MHz, CDCl₃) δ 2.47-2.41 (m, 3H), 2.27- 2.19 (m, 2H), 2.10 (d, J= 12.8 Hz, IH), 2.02-1.86 (m, 4H), 1.76-1.16 (m, 6H), 1.18 (s, 9H), 0.92 (d, J= 6.0 Hz, 3H), 0.61 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.19, 211.94, 61.88, 56.59, 49.82, 44.18, 40.89, 38.89, 35.11, 33.20, 29.80, 27.37, 26.41, 23.97, 19.02, 18.51, 12.45; IR (neat, cm^"1) 2958, 2872, 1708, 1474, 1364, 1223, 1068; HRMS m/z [M + Na⁺] calcd 315.2294 for C₁₉H₃₂O₂Na⁺, found 315.2276.

[0136] B. Preparation of 24(O)TB analog (+)-3. Enantiomerically pure A-ring phosphine oxide (-)-31 and C,D-ring ketone (+)-29, each was separately azeotropically dried with anhydrous benzene (4x 1 mL) on a rotary evaporator and held under vacuum (ca. 0.1 mmHg) for at least 48 hours prior to use. A flame-dried 10-mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon was charged with enantiomerically pure A-ring phosphine oxide (-)-31 (50 mg, 0.08 mmol) which was dissolved in 2 mL freshly distilled THF. The flask was cooled down to -78 ⁰C in a dry ice bath. To this solution, n-BuLi (48 μL, 0.08 mmol, 1.6 M solution inhexanes) was added dropwise over several minutes, during which time a deep red color was developed and persisted. This mixture was allowed to stir at -78 ⁰C for an additional 10 minutes. Meanwhile, a flame-dried 10-mL recovery flask equipped with a magnetic stir bar, and a septum along with an Ar balloon was charged with enantiomerically pure C,D-ring ketone (+)-29 (22 mg, 0.075 mmol), dissolved in 1 mL freshly distilled THF, and cooled down to - 78 ⁰C in an dry ice bath. The solution of C,D-ring ketone was gently transferred into the flask containing the phosphine oxide anion at -78 ⁰C via cannula over several minutes. After the addition was complete, the deep red color persisted and the mixture was allowed to stir at -78 ⁰C for 9 hours, during that time it was visually checked. On observation of the light yellow color, the reaction was quenched at -78 ⁰C by adding 3 mL of pH 7 buffer and allowed to come to room temperature. The mixture was then rinsed into a separatory funnel with ethyl acetate and extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL) and brine solution, dried over MgSO₄, filtered, and purified using silica gel chromatography (5% ethyl acetate/hexanes) to give a colorless oil coupled product (13 mg, 35%). A 15-mL round-bottomed flask was charged with the coupled 5 product (13 mg, 0.02 mmol) dissolved in 5 mL of distilled THF. Tetrabutylammonium fluoride (TBAF 1 M solution in THF, 59 μL, 0.06 mmol) was added to the reaction flask, and this solution was left to stir at room temperature for 7 hours. The resulting mixture was quenched with 2 mL of water, extracted with ethyl acetate (3x 25 mL), dried over MgSO₄, concentrated, and purified using silica gel column chromatography (25% ethyl 0 acetate/hexanes) to give 24(O)TB 3 (7 mg, 82%) as a colorless oil. The product was separated by an HPLC using Chiral OD [semipreparative (Ix 25 cm), t_R = 38.5 min] eluted with 7% IPA in hexanes to afford 4 mg of 24(O)TB 3 (compound 3): [α£⁵ +29.6 (c 3.0, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.36 (d, J= 11.2 Hz, IH), 5.99 (d, J= 11.2 Hz, IH), 5.32 (s, IH), 4.99 (s, IH), 4.42 (m, IH), 4.22 (m, IH), 2.81 (dd, J = 12.0, 4.0 Hz, IH), 2.59 5 (dd, J= 12.0, 2.8 Hz, IH), 2.52-2.28 (m, 5H)₅ 2.20-1.20 (m, 16H), 1.13 (s, 9H), 0.91 (d, J= 6.4 Hz, 3H), 0.53 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 216.5, 147.5, 143.1, 132.9, 124.9, 117.0, 111.8, 70.8, 66.8, 56.4, 56.2, 45.8, 45.2, 44.2, 42.8, 40.4, 35.7, 33.2, 30.2, 29.0, 27.5, 26.4, 23.5, 22.2, 18.6, 14.6, 12.0; IR (neat, cm^"1) 3356, 2949, 2872, 1703, 1465, 1364, 1054, 754; HRMS m/z [M + Na⁺] calcd 451.3182 for C₂₈H₄₄O₃Na⁺, found 451.3178. 0

EXAMPLE 4 23-Enone analogs (+>4a and 4b (Figure 19)

[0137] A. Preparation of Cyano-C,D-ring ketone 35. A flame-dried 10-mL recovery 5 flask equipped with a magnetic stir bar, and a septum along with an Ar balloon was charged with C-8 alcohol derived from C-8 silyl ether 33 (50 mg, 0.22 mmol) dissolved in 2.2 mL freshly distilled CH₂Cl₂. To this solution, PDC (170 mg, 0.45 mmol) and 0.22 g of oven- dried Celite were added in one portion at room temperature. The resulting mixture was allowed to stir at room temperature for about 12 hours. The mixture was directly purified 0 by column chromatography eluted with first 20% ethyl acetate in hexanes to afford 43 mg of ketone 35 in 83% yield: ¹H NMR (CDCl₃, 400 MHz) δ 2.52 (dd, J= 7.6 Hz, IH), 2.41- 2.21 (m, 4H), 2.12-2.01 (m, 2H), 1.99-1.74 (m, 4H), 1.69-1.54 (m, 2H), 1.37- 1.25 (m, IH), 1.2 (d, J= 6.8 Hz, 3H), 0.67 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) 8 211.1, 118.4, 61.5, 54.9, 49.5, 40.7, 38.4, 33.1, 27.3, 24.6, 23.7, 19.2, 18.9, 12.5; HRMS m/z [M + Na⁺] calcd 242.1515 for C₁₄H₂₁NO, found 242.1519.

[0138] B. 23-Enone analogs (+)-4a and 4b. Phosphine oxide (±)-31 and C,D-ring ketone 35, each was separately azeotropically dried with anhydrous benzene (4x 1 niL) on a rotary evaporator and held under vacuum (ca. 0.1 mrnHg) for at least 48 hours prior to use. A flamedried 10-mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon was charged with phosphine oxide (±)-31 (60 mg, 0.1 mmol) and dissolved in ca 1.2 mL freshly distilled THF to give ca. 0.1 M solution, and the flask was cooled down to -78 ⁰C in a dry ice bath. To this solution, n-BuLi (70 μL, 0.11 mmol, 1.6 M solution in hexanes) was added dropwise over several minutes, during which time a deep red color was developed and persisted. This mixture was allowed to stir at -78 ⁰C for an additional 10 minutes. Meanwhile, a flame-dried 10-mL recovery flask equipped with a magnetic stir bar, and a septum along with an Ar balloon was charged with C,D-ring ketone 35 (15 mg, 0.068 mmol), dissolved in 0.5 mL freshly distilled THF, and cooled down to -78 ⁰C in a dry ice bath. The solution of C,D-ring ketone 35 was transferred dropwise into the flask containing the phosphine oxide anion at-78⁰C via cannula over several minutes. After the addition was complete, the deep red color persisted and the mixture was allowed to stir at - 78 ⁰C for 2 hours, during that time it was visually checked. On observation of the light yellow color, the reaction was quenched at -78 ⁰C by adding of 3 mL of pH7 buffer and allowed to come to room temperature. The mixture was then rinsed into a separatory funnel with ethyl acetate and extracted with ethyl acetate (4x 25 mL). The combined extracts were washed with water (25 mL) and brine solution (25 mL), dried over Na₂SO₄, and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by column chromatography eluted with 25% ethyl acetate in hexanes in the presence of 1% triethylamine to afford 34 mg of the coupled product in 85% yield in a 3.8:1 ratio as determined by ¹H NMR. This coupled product (30 mg, 0.051 mmol) was charged into 5- rtiL round-bottomed flask equipped with a magnetic stir bar, and a septum along an Ar balloon and dissolved in 1 mL anhydrous dichloromethane to give ca. 0.05 M solution. Then the flask was cooled down to -78 ⁰C. To this well-stirred solution, DIBAL (0.077 mmol, 51.6 mL, 1.5 M solution in toluene) was added via syringe at this temperature and the mixture was then allowed to stir at -78 ⁰C for 1 hour. This reaction mixture was diluted with ether (25 mL), and 1 N solution of HCl (ca. 1 mL) was added and stirred for a few minutes. The reaction mixture was then rinsed into a separatory funnel with ethyl acetate and was extracted with ethyl acetate (4x 10 mL). The combined extracts were washed with US2006/000183

water (10 mL) and brine solution (10 niL), dried over Na₂SO₄, and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by flash column chromatography eluted with 5% ethyl acetate in hexanes in the presence of 1% triethylamine affording 25 mg of 23-aldehyde 34 in 83% yield. A flame-dried 10-mL recovery flask equipped with a magnetic stir bar, and a septum along with an Ar balloon was charged with KOtBu (3.5 mg, 0.031 mmol) and 0.5 mL freshly distilled THF. Then, the flask was cooled down to -78 ⁰C in a dry ice bath. To this solution, dimethylphosphonate 36 (6.4 mg, 0.031 mmol) was added as a solution in 0.5 mL freshly distilled THF. After 30 minutes, 0.5 mL of THF solution of aldehyde 34 (15 mg, 0.026 mmol) was transferred into the flask via cannula over several minutes at -78 ⁰C. After the addition was complete, the mixture was gradually warmed up to room temperature and then stirred for about 4 hours. The reaction was quenched by adding 2 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 10 mL). The combined extracts were washed with water (10 mL), and brine solution (10 mL), dried over Na₂SO₄, and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by flash column chromatography eluted with 5% ethyl acetate in hexanes to afford 12 mg of trans-oleum as determined by ¹H NMR in 70% yield. This product (12 mg, 0.018 mmol) was charged into a 5 mL argon purged polypropylene vial equipped with a magnetic stir bar, a septum along with a cap and dissolved in 1.O mL acetonitrile to give ca. 0.02 M solution. To this well-stirred solution, HF (1.8 mmol, 49% aqueous solution) was added via syringe at room temperature and the mixture was then allowed to stir at room temperature for 2 hours. TLC showed the completion of the reaction. This reaction mixture was diluted with ether (25 mL) and saturated solution OfNaHCO₃ was added until no more carbon dioxide was liberated. The reaction mixture was then rinsed into a separatory funnel with ethyl acetate and was extracted with ethyl acetate (4x 10 mL). The combined extracts were washed with water (10 mL) and brine solution (10 mL), dried over Na₂SO₄, and filtered. The filtrate was concentrated in vacuo to give the crude product as a mixture of diastereomers in 4:1 ratio (as determined by ¹H NMR of the crude reaction mixture). The crude was purified by column chromatography eluted with 100% ethyl acetate in the presence of 1% TEA to afford 5.2 mg of a diastereomeric mixture of 4a and 4b in 66% yield. This analog 4b was purified by an HPLC [semipreparative (Ix 25 cm) Chiracel OD, t_R = 48.2 min] eluted with 5% isopropyl alcohol in hexanes to afford 2.2 mg of 4b: ¹H NMR (CDCl₃, 400 MHz) δ 6.95-6.88 (m, IH), 6.49 (d, J= 15.2 Hz, IH), 6.37 (d, J= 11.2 Hz, IH), 6.01 (d, J= 11.6 Hz₅ IH), 5.33-5.32 (m, IH), 5.00-4.99 (m, IH), 4.45-4.01 (m, IH), 4.25-4.22 (m, IH), 2.84 (dd, J= 12.0, 4.4 Hz, IH), 2.60 (dd, J= 13.2, 5.0 Hz, IH), 2.34- 2.29 (m, 2H), 2.07- 1.89 (m, 6H), 1.68-1.49 (m, 17H), 1.35-1.25 (m, 4H), 0.95 (d, J= 6.4 Hz, 3H), 0.55 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 204.3, 147.6, 146.2, 142.9, 132.9, 125.7, 124.9, 117.1, 111.8, 70.8, 66.8, 56.2, 56.0, 45.9, 45.2, 42.8, 42.7, 40.3, 39.3, 36.0, 29.0, 27.6, 26.2, 23.5, 22.2, 19.1, 12.0; IR (neat, cm^"1) 3369, 2949, 2872, 1684, 1619, 1437, 1366, 1055, 908; HRMS m/z [M + Na⁺] calcd 463.3183 for C₃₀H₄₅NO₃Na⁺, found 463.3193; UV (MeOH) X_1113x = 265 nm (6 = 15,864).

EXAMPLE 5

22-Ene-24(O1TB (+1-5 (Figure 20)

[0139] A. Preparation of 22-Ene-24 ketone silyl ether 38. To a solution of the phosphate 36 (80 mg, 0.38 mmol) in THF (5 niL), potassium tert-butoxide (43 mg, 0.38 mg) was added at 0 ⁰C. After stirring for 1 hour at 0 ⁰C, a solution of the aldehyde (+)-37 (60 mg, 0.18 mmol) in THF (2 mL) was added via cannula at room temperature. Then, the mixture was stirred for 4 days at room temperature. The resulting mixture was quenched with water (3 mL), extracted with EtOAc (3x 25 mL), dried over MgSO₄, and concentrated. The residue was subjected to column chromatography with EtOAc/hexanes (1 : 15) as eluent to afford 50 mg (68%) of the desired ketone 38 as a colorless oil: [αj? +73.4 (c 1.0,

CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.75 (dd, J= 15.2, 9.2 Hz, IH), 6.39 (dd, J= 15.2, 0.8 Hz, IH), 4.03 (m, IH), 2.24 (m, IH), 1.93 (dt, J= 12.4, 2.8 Hz, IH), 1.82 (m, IH), 1.50-1.69 (m, 4H), 1.13- 1.41 (m, 6H), 1.14 (s, 9H), 1.05 (d, J= 6.4 Hz, 3H), 0.94 (s, 3H), 0.935 (t, J= 8.0 Hz, 9H), 0.54 (q, J= 8.0 Hz, 6H); ¹³C NMR (100 MHz, CDCl₃) δ 204.8, 152.9, 121.8, 69.3, 55.5, 52.9, 42.8, 42.4, 40.6, 39.7, 34.6, 27.4, 26.2, 23.0, 19.2, 17.7, 13.8, 6.9, 4.9; IR (neat, cm^"1) 2953, 2875, 1725, 1690, 1623, 1457, 1366, 1234, 1166, 1081, 1018, 725; HRMS m/z [M + Na⁺] calcd 429.3159 for C₂₅H₄₆O₂SiNa⁺, found 429.3161.

[0140] B. Preparation of 22-Ene-24 ketone C,D-ring ketone 40. To a solution of silyl ether 38 (46 mg, 0.11 mmol) in THF (3 mL), 0.35 mL (0.35 mmol) of a 1.0 M solution of TBAF in THF was added at room temperature, and then it was stirred overnight at room temperature. The reaction mixture was quenched with water (2 mL), extracted with EtOAc (3x, 20), washed with brine, dried over MgSO₄, and concentrated. The residue was subjected to column chromatography with EtOAc/hexanes (1 :3) as eluent to give 12 mg (37%) of the desired alcohol as a colorless oil. To a solution of the alcohol (12 mg, 0.041 mmol) in CH₂Cl₂ (4 mL), 120 mg of oven-dried Celite and PDC (120 mg, 0.33 mmol) was added at room temperature. The mixture solution was stirred overnight and then passed through a 2 cm pad of flash silica gel and washed with EtOAc. The filtrate was 5 concentrated and subjected to column chromatography with EtOAc/hexanes (1 :4) as eluent

CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.74 (dd, J= 15.2, 9.2 Hz, IH)₅ 6.42 (dd, J= 15.2, 8.0 Hz, IH), 2.46 (dd, J= 11.2, 7.6 Hz, IH), 2.18-2.32 (m, 3H), 2.00-2.12 (m, 2H), 1.91 (m, IH), 1.46-1.78 (m, 5H), 1.27 (m, IH), 1.14 (s, 9H), 1.12 (d, J= 6.4 Hz, 3H), 0.67 (s, 10 3H); ¹³C NMR (100 MHz, CDCl₃) δ 211.5, 204.6, 151.5, 122.3, 61.7, 55.3, 49.9, 42.9, 40.9, 39.7, 38.8, 27.5, 26.2, 24.0, 19.5, 19.1, 12.7; IR (neat, cm^'1) 2961, 2872, 1713, 1688, 1622, 1477, 1366, 1233, 1076, 989, 950, 860; HRMS m/z [M + Na⁺] calcd 313.2138 for C₁₉H₃₀O₂Na⁺, found 313.2132.

[0141] C. Preparation of 22-Ene-24(O)TB (+)-5. A solution of 50 mg (0.086 mmol) of

15 enantiomerically pure phosphine oxide (-)-31 in 1.5 mL of anhydrous THF was cooled to - 78 ⁰C and treated with 54 μL (0.086 mmol, 1.6 M in hexanes) of π-BuLi under argon atmosphere. The mixture turned deep reddish color and was stirred for 15 min at -78 ⁰C. To the solution, a precooled (-78 ⁰C) solution of 10 mg (0.034 mmol) of the enantiomerically pure C,D-ring ketone (+)-40 in 1.5 mL of anhydrous THF via cannula was

20 added dropwise. The reaction kept going until the reddish-orange color faded to yellow

(about 3 hours). The reaction was quenched by adding 1.0 mL of pH 7 buffer at -78 ⁰C, and then warmed to room temperature, extracted with EtOAc (3x 20 mL), washed with brine, dried over MgSO₄, and concentrated. The residue was subjected to column chromatography with EtOAc/hexanes (1:10) as eluent to afford 4 mg (18%) of the coupled product as a

25 colorless oil. The coupled product (4 mg, 0.0060 mmol) was dissolved in 2 mL of anhydrous EtOH, and to the solution, 50 μL of 49% aq HF was added. The resulting mixture was stirred 2 hours at room temperature, and then quenched with 5 mL of satd NaHCO₃ solution. The solution was stirred for 10 min, and then extracted with EtOAc (3x 20 ml), washed with brine, dried over MgSO₄, and concentrated. The residue was subjected

30 to column chromatography with EtOAc as eluent to give 2 mg (93%) of the desired product

5 as a colorless oil: [αg +65.0 (c 0.10, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.78 (dd, J = 15.2, 9.2 Hz, IH), 6.41 (d, J= 15.2 Hz, IH), 6.37 (d, J= 11.2 Hz, IH), 6.01 (d, J= 11.2 Hz, IH), 5.32 (m, IH), 4.99 (m, IH), 4.43 (m, IH), 4.23 (m, IH), 2.83 (dd, J= 11.6, 3.2 Hz, IH), 2.60 (dd, J= 13.2, 3.2 Hz, IH)₅ 2.20-2.34 (m, 2H), 1.89-2.04 (m, 4H), 1.43-1.76 (m, 6H), 1.14-1.41 (m, 5H), 1.15 (s, 9H), 1.09 (d, J= 6.8 Hz, 3H), 0.58 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 189.1, 152.4, 147.6, 142.6, 133.1, 124.9, 122.1, 117.2, 111.8, 70.8, 66.8, 56.1, 55.3, 46.1, 45.3, 42.8, 40.3, 40.2, 29.7, 29.0, 27.4, 26.2, 23.5, 22.3, 19.6, 12.3; IR 5 (neat, cm^"1) 3401, 2955, 2926, 2872, 1684, 1653, 1617, 1558, 1546, 1507, 1457, 1079.

[0142] The product 5 was further purified by HPLC (t_R = 28.2 minutes, Chiral OD column, 7% 2-proρanol in Hexanes, 2.5 mL/min, 254 nm) for testing the biological data.

EXAMPLE 6 24-F2-25(OKB analog (-V6 (Figure 21)

10

[0143] A. Preparation of Difluoro ethyl ester 42. A suspension of activated zinc dust (98.5 mg, 1.51 mmol) and ethyl bromodifluoroacetate (0.19 mL, 1.51 mmol) in THF (5 mL) was refluxed for 20 min and then cooled to 0⁰C. To this, the solution of the aldehyde (+)- 39 (100 mg, 0.30 mmol) in THF (5 mL) was added. The reaction mixture was warmed to

15 room temperature, followed by refluxing for 20 min, and then cooled to room temperature. The reaction mixture was poured into 1 M KHSO₄ and extracted with EtOAc (3x 20 mL), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (10% EtOAc/hexanes) to give 84.5 mg (62%) of a 1:1 mixture of diastereomers of the desired alcohol as a colorless oil. To a solution of ethyl ester (84 mg,

20 0.18 mmol) and pyridine (0.066 mL, 0.82 mmol) in CH₂Cl₂ (5 mL), phenyl chlorothionoformate (0.053 mL, 0.38 mmol) was added. After being stirred at room temperature for 20 hours, the reaction mixture was diluted with water, extracted with ether (3x 20 mL), washed with satd NaHCO₃ solution and brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (5% EtOAc/hexanes) to give 97.8

25 mg (90%) of the desired carbonate as diastereomeric mixtures. To the solution of the resulting phenylthianocarbonate (97.5 mg, 0.16 mmol) in anhydrous benzene (10 mL), 2,2'- azobisisobutyronitrile (AlBN, 5 mg) and tributyltin hydride (0.066 mL, 0.24 mmol) were added at room temperature. After being refluxed for 3 hours, the mixture was cooled to 0 ⁰C, diluted with water, extracted with EtOAc (3x 20 mL), washed with brine, dried over

30 MgSO₄, concentrated in vacuo, and then purified by column chromatography (3%

EtOAc/hexanes) to give 58.2 mg (80%) of the desired difluoro ester 42 as a colorless oil: ¹H NMR (400 MHz, CDCl₃) δ 4.32 (q, J= 7.2 Hz, 2H), 4.03 (s, IH), 2.14-2.06 (m, IH), 1.98- 1.89 (m, 2H), 1.85-1.75 (m, 2H), 1.69-1.64 (m, IH), 1.61-1.53 (m, 2H), 1.49-1.43 (m, IH), 1.38-1.34 (m, 2H), 1.35 (t, J= 7.2 Hz₅ 3H), 1.26-1.17 (m, 3H), 1.13-0.99 (m, 2H), 0.94 (t, J= 8.0 Hz, 9H), 0.91 (d, J= 6.0 Hz₅ 3H), 0.90 (s, 3H), 0.55 (q, J= 8.0 Hz, 6H).

[0144] Preparation of Difluoro-C,D-ring diketone 44. To a solution of ethyl ester 42 (54 mg, 0.12 mmol) in THF (5 mL) was added 125.1 μL of tert-BuLi (1.43 M in pentane, 0.18 mmol) at -78⁰C. After being stirred for 3 hours, the reaction mixture was warmed up to room temperature, diluted with water, extracted with ether (3x 20 mL), washed with satd NaHCO₃ solution and brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (hexanes only) to give 41.6 mg (75%) of the desired ketone as a colorless oil: ¹HNMR (400 MHz₅ CDCl₃) δ 4.02 (d, J= 2.4 Hz, IH)₅ 1.95-1.90 (m, IH), 1.87-1.74 (m, 2H), 1.69-1.64 (m, IH), 1.57-1.52 (m, 2H), 1.49-1.43 (m, 2H), 1.39- 1.31 (m, 3H), 1.26 (s, 9H), 1.24-1.20 (m, 2H), 1.12- 0.99 (m, 2H), 0.94 (t, J= 8.0 Hz, 9H), 0.89 (d, J= 6.0 Hz₅ 3H), 0.89 (s, 3H), 0.55 (q, J= 8.0 Hz, 6H). To a solution of ketone (30 mg, 0.065 mmol) in THF (5.0 mL), 0.20 mL (0.20 mmol) of a 1.0 M solution of TBAF in THF was added, and then it was stirred at 0⁰C for 1 hour and stored overnight at room temperature. The reaction mixture was quenched with water (5 mL), extracted with EtOAc (3x 20 mL), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (25% EtOAc/hexanes) to give 21.0 mg (93%) of alcohol as a colorless oil. To a solution of the C,D-ring alcohol (14.1 mg, 0.041 mmol) in CH₂Cl₂ (5 mL) were added 50 mg of oven-dried Celite and PDC (46.2 mg, 0.12 mmol) at room temperature. The reaction mixture was stirred overnight and then passed through a 2 cm pad of flash silica gel and washed with EtOAc. The filtrate was concentrated and purified by column chromatography (20% EtOAc/hexanes) to give 13.0 mg (93%) of the desired

C₅D-ring diketone 44 as a colorless oil: [α£⁵ +1.30 (c 1.01, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 2.45 (dd, J= 11.6, 7.6 Hz, IH), 2.30-2.21 (m, 2H), 2.11- 2.07 (m, 2H), 2.05-1.97 (m, IH), 1.96-1.84.(m, 3H), 1.76-1.71 (m, IH), 1.61-1.42 (m, 5H), 1.33-1.21 (m, 2H), 1.25 (s, 9H)₅ 0.97 (d, J= 6.0 Hz, 3H), 0.64 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 211.83, 205.23 (J= 15.5 Hz), 120.74 (J= 253.1 Hz), 61.87, 55.99, 49.81, 43.34, 40.90, 38.89, 34.91, 30.54 (J= 22.7 Hz), 27.30, 27.04 (J= 4.1 Hz), 26.07 (J= 2.3 Hz)₅ 23.99, 19.01, 18.41, 12.47; IR (neat, cm^"1) 2954, 2860, 1719, 1460, 1366, 1307, 1231, 1196, 1043, 1008, 967, 943, 914, 843, 773, 737; HRMS m/z [M + Na⁺] calcd 365.2262 for C₂₀H₃₂F₂O₂Na⁺, found 365.2252. ,

[0145] C. Preparation of 24-F2-25(O)TB analog (-)-6. To a solution of 50.0 nig (0.086 mmol) of enantiomerically pure A-ring phosphine oxide (-)-31 in 2.0 niL of anhydrous THF, 53.6 μL (0.086 mmol, 1.6 M in hexanes) of n-BuLi was added at -78 ⁰C, and then the reddish solution was stirred for 15 min at -78 ⁰C. To the solution, a precooled

5 (-78 ⁰C) solution of enantiomerically pure C,D-ring ketone (+)-44 (8.0 mg, 0.023 mmol) in 1.0 mL of anhydrous THF was added dropwise. The reaction kept going until the reddish- orange color faded to yellow (about 6 h). The reaction was quenched by adding 1.0 mL of pH 7 buffer, and then warmed up to room temperature, extracted with EtOAc (2x 20 mL), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column

10 chromatography (20% EtOAc/hexanes) to afford 13.3 mg (81%) of the coupled product as a pale yellow oil. The coupled product (13.2 mg, 0.019 mmol) was dissolved in 5 mL of anhydrous THF, and to this solution, 0.075 mL (0.075 mmol) of a 1.0 M solution of TBAF in THF was added. The reaction was run in darkness overnight, and then extracted with EtOAc (2x 30 mL), washed with brine, dried over MgSO₄, concentrated in vacuo, and then

15 purified by column chromatography (5% MeOH/CH₂Cl₂) to give 8.48 mg (95%) of the desired compound 6 as a colorless oil: [αg -23.3 (c 0.42, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.37 (d, J= 11.6 Hz, IH), 6.01 (d, J= 11.2 Hz, IH), 5.32 (t, J= 1.6 Hz, IH), 5.00 (t, J= 1.6 Hz, IH), 4.44-4.41 (m, IH), 4.24-4.21 (m, IH), 2.82 (dd, J= 12.8, 4.4 Hz, IH), 2.60 (dd, J= 13.6, 3.6 Hz, ¹H), 3.34-2.28 (m, IH), 2.07-1.82 (m, 6H), 1.70-1.58 (m, 5H),

20 1.55-1.42 (m, 5H), 1.31-1.27 (m, 2H), 1.26 (s, 9H) 0.94 (d, J= 6.4 Hz, 3H), 0.54 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 205.33 (J= 29.1 Hz), 147.58, 142.95, 132.96, 124.93, 120.86 (J= 253.6 Hz), 117.09, 111.81, 70.82, 66.84, 56.26, 55.86, 45.85, 45.24, 43.34, 42.82, 40.39, 35.50, 30.64 (J= 22.7 Hz), 29.03, 27.39, 27.13, 26.08, 23.52, 22.19, 18.51, 11.98; IR (neat, cm^'1) 3365, 2942, 2860, 1725, 1642, 1454, 1366, 1225, 1049, 961, 908,

25 796, 749, 737, 596; UV (MeOH) W = 252 nm (e= 3166).

EXAMPLE 7 24-fCH3)2-25(0yTB analog (+V7 (Figure 2D

30 [0146] A. Preparation of Ester 43. Lithium diisopropylamide (LDA) solution was prepared by treating diisopropylamine (213.5 mg, 2.11 mmol) in THF (5 mL) at -78 ⁰C with a 1.6 M solution of n-BuLi in hexanes (1.3 mL, 2.08 mml). The LDA solution was stirred at -78 ⁰C for 30 min, and then a solution of ethyl isobutyrate (245 mg, 2.11 mmol) in THF (2 mL) was added. After being stirred for 1 hour at -78 ⁰C, a solution of iodide (+)-22 (95 mg, 0.211 mmol) in THF (2 mL) was added. The reaction mixture was stirred at -78 ⁰C for 2 hours, then warmed up to room temperature and quenched with the addition of water (5 mL). The reaction mixture was extracted with diethyl ether (4x 15 mL). The organic layer 5 was washed with water and brine, dried over sodium sulfate, concentrated in vacuo, and then purified by column chromatography (5% EtOAc/petroleum ether) to give 91.8 mg of ester 43 (0.209 mmol, 99% yield) as a colorless oil: [αg +41.8 (c 1.50, CHCl₃); ¹HNMR (CDCl₃, 400 MHz) δ 4.104 and 4.102 (q, J= 7.2 Hz, 2H), 4.03-4.02 (m, IH), 1.93 (dm, J= 12.4 Hz, IH), 1.83-1.72 (m, 2H), 1.67-1.65 (m, IH), 1.61-1.50 (m, 5H), 1.43-1.27 (m,

10 6H), 1.24 (t, J= 7.2 Hz, 3H), 1.19- 0.97 (m, 2H), 1.14 (s, 3H), 1.137 (s, 3H), 0.94 (t, J= 8.0 Hz, 9H), 0.88 (s, 3H), 0.87 (d, J= 6.4 Hz, 3H), 0.55 (q, J= 8.0 Hz, 6H); ¹³C NMR (CDCl₃, 100MHz) δ 178.12, 69.38, 60.10, 56.28, 53.07, 42.15, 42.07, 40.74, 36.97, 35.40, 34.65, 30.61, 27.11, 25.25, 25.11, 22.99, 18.63, 17.67, 14.28, 13.46, 6.93, 4.93; IR (neat, cm ¹) 2950, 2875, 1731, 1472, 1144, 1025, 742; HRMS m/z [M + Na⁺] calcd 461.3421 for

15 C₂₆H₅₀O₃SiNa⁺, found 461.3398.

[0147] B. Preparation of Diketone 45. To a solution of ester 43 (90 mg, 0.205 mmol) in diethyl ether (4 mL), a solution of tert-butyllithium (1.43 M in pentane, 0.15 mL, 0.215 mmol) was added dropwise at -78 ⁰C. After being stirred for 1 hour at -78 ⁰C, the reaction mixture was warmed up to room temperature, and then quenched with the addition of water 20 (4 mL). The reaction mixture was extracted with diethyl ether (4x 15 mL). The organic layer was washed with water and brine, dried over sodium sulfate, concentrated in vacuo, and then purified by column chromatography (5% EtO Ac/petroleum ether) to give 83.1 mg of tertbutyl ketone (0.184 mmol, 90% yield) as a colorless oil: [α]" +41.1 (c 0.71, CHCl₃); ¹HNMR (CDCl₃, 400 MHz) δ 4.02-4.01 (m, IH), 1.93 (dm, J= 12.8 Hz, IH), 1.83-1.72

25 (m, 2H), 1.67-1.42 (m, 5H), 1.38-0.99 (m, 7H), 1.37-1.14 (m, 2H), 1.23 (s, 3H), 1.22 (s, 9H), 1.19 (s, 3H), 0.94 (t, J= 8.0 Hz, 9H), 0.88 (s, 3H), 0.87 (d, J= 6.4 Hz, 3H), 0.54 (q, J = 8.0 Hz, 6H); ¹³C NMR (CDCl₃, 100 MHz) δ 218.85, 69.39, 56.49, 53.06, 49.48, 45.66, 42.08, 40.74, 38.08, 35.90, 34.63, 30.99, 28.45, 27.31, 26.63, 26.52, 23.01, 18.55, 17.67, 13.45, 6.93, 4.93; IR (neat, cm^'1) 2952, 2876, 1684, 1476, 1367, 1166, 1085, 1019, 725;

30 HRMS m/z [M + Na⁺] calcd 473.3785 for C₂₈H₅₄O₂SiNa⁺, found 473.3873.

[0148] To a solution of tert-butyl ketone (40 mg, 0.0887 mmol) in THF (3 mL), a solution of tetrabutylammonium fluoride (TBAF, 1.0 M in THF, 0.89 mL, 0.89 mmol) was added. ₍

After being stirred for 4 hours at room temperature, the reaction mixture was concentrated. Purification of the residue by flash column chromatography (20% EtOAc/hexanes) provided 29.6 mg (0.088 mmol, 99% yield) of the corresponding alcohol. A solution of this alcohol (29.6 mg) in CH₂Cl₂ (3 mL) was added to pyridinium dichromate (PDC, 66.7 mg, 0.177 5 mmol, 2 equiv) and Celite (70 mg) at room temperature under argon atmosphere. After being stirred overnight, the reaction mixture was diluted with EtOAc and filtered through a silica gel plug. The filtrate was concentrated in vacuo and then purified by column chromatography (20% EtO Ac/petroleum ether) to give 27.6 mg of the diketone 45 (0.0825 mmol, 94% yield) as colorless oil: [α{? +8.7 (c 1.33, CHCl₃); ¹H NMR (CDCl₃, 400 MHz) 0 δ 2.42 (dd, J= 11.4, 7.4 Hz, IH)₅ 2.29-2.16 (m, 2H), 2.09 (dm, J= 13.2 Hz, IH), 2.03-1.81 (m, 3H), 1.76-1.39 (m, 8H), 1.37-1.14 (m, 2H), 1.23 (s, 3H), 1.21 (s, 9H), 1.20 (s, 3H), 0.94 (d, J= 6.4 Hz, 3H), 0.61 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 218.68, 212.04, 61.93, 56.27, 49.84, 49.34, 45.70, 40.93, 38.91, 38.12, 36.01, 30.96, 28.36, 27.46, 26.59, 26.55, 24.02, 19.05, 18.66, 12.41; IR (neat, cm ¹) 2957, 2873, 1715, 1682, 1477, 1385, 5 1366, 1220, 1044, 982; HRMS m/z [M + Na⁺] calcd 357.2764 for C₂₂H₃₈O₂Na⁺, found 357.2750.

[0149] C. Preparation of 24-(CH3)2-25(O)TB analog (+)-7. Phosphine oxide (-)-31 and enantiomerically pure C,D-ring diketone 45 were separately azeotropically dried four times with benzene and held under vacuum for 60 hours immediately prior to use. To a 0 solution of phosphine oxide (-)-31 (62.0 mg, 0.106 mmol) in THF (2 mL), a 1.6 M solution of n-BuLi in hexanes (66 μL, 0.106 mmol) was added dropwise at -78 ⁰C under argon atmosphere. The resulting deep reddish-orange solution was allowed to stir for 20 min, at which time a precooled (-78 ⁰C) solution of enantiomerically pure C,D-ring diketone (+)-45 (15.8 mg, 0.0472 mmol) in THF (2 mL) was transferred dropwise via cannula. The deep 5 reddish-orange solution was stirred in the dark for 5 hours, during which time the color was faded. On observation of a light yellow color, the reaction mixture was quenched at -78 ⁰C with 3 mL of buffer water (pH 7). The mixture was allowed to come up to room temperature, extracted with EtOAc (4x 10 mL), dried over Na₂SO₄, filtered, concentrated, and purified by silica gel column chromatography (5% EtO Ac/petroleum ether) to give 17

30 mg of the coupled product (0.0243 mmol, 52% yield). This coupled product was dissolved in THF (2 mL) and treated with a solution of tetrabutylammonium fluoride (TBAF, 1.0 M in THF, 0.24 mL, 0.24 mmol). After being stirred overnight at room temperature, the reaction mixture was concentrated. Purification of the residue by flash column chromatography (100% EtOAc) provided 9.9 mg of 24-(CH₃)2-25(O)TB 7 (0.021 mmol, 86% yield) as a white solid: mp (⁰C) 109-112; [αg +40.7 (c 0.48, CHCl₃); ¹H NMR

(CDCl₃, 400 MHz) δ 6.37 (d, J= 11.2 Hz, IH), 6.01 (d, J= 11.2 Hz, IH), 6.07 (d, J= 16.0 Hz, IH), 5.32 (m, IH), 5.00 (m, IH), 4.43 (m, IH), 4.24-4.22 (m, IH), 2.81 (dd, J= 12.0, 4.0 Hz, IH), 2.59 (dd, J= 13.2, 3.2 Hz, IH), 2.31 (dd, J= 13.4, 6.6 Hz, IH), 2.05-1.81 (m, 5H), 1.72-1.43 (m, 10H), 1.32-1.19 (m, 4H), 1.23 (s, 3H), 1.22 (s, 9H), 1.21 (s, 3H), 0.91 (d, J= 6.0 Hz, 3H), 0.52 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 218.85, 147.60, 143.18, 132.81, 124.99, 116.99, 111.77, 70.82, 66.85, 56.30, 56.17, 49.45, 45.86, 45.25, 42.83, 40.41, 38.10, 36.64, 31.06, 29.06, 28.43, 27.58, 26.66, 26.51, 23.57, 22.25, 20.70, 18.78, 11.95; IR (neat, cm^"1) 3350, 2949, 2872, 1682, 1475, 1054, 755; HRMS m/z [M + Na⁺] calcd 493.3652 for C₃₁H₅₀O₃Na⁺, found 493.3675; UV (MeOH) )w = 265 nm (e = 14,822).

Example 8

25(OVMe₂Ph 8 (Figure 22)

[0150] A. Preparation of TES ether 48. In a flame-dried, 25 -mL, round-bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed diisoproylamine (0.070 mL, 0.50 mmol) in THF (1 mL). The solution was cooled to -78 ⁰C followed by the addition of n-BuLi (0.31 mL, 1.6 M in hexane, 0.50 mmol), and was stirred for 10 minutes. To this was added a solution of 3-methyl-3- phenylbutan-2-one (0.081 g, 0.50 mmol) in THF (1 mL), and the resulting mixture was stirred for 30 min followed by the addition of hexamethylphosphoramide (0.20 mL). The reaction mixture was stirred for an additional 15 min, and to this was added a solution of iodide 22 (0.030 g, 0.067 mmol) in THF (1 mL). The mixture was slowly warmed to room temperature and stirred for 16 hours. Then, it was quenched with H₂O (3 mL) and extracted with EtOAc (3x 3 mL). The combined organic phases were washed with H₂O (Ix 3 mL) and brine (Ix 3 mL), dried (MgSO₄), and concentrated. Flash chromatography (EtOAc :hexane,

5:95) afforded 0.028 g (85%) of 48 as an oil; [α£⁵ = +38.6 (c 0.84, CHCl₃); IR (neat cm^'1) 2961, 1701. ¹HNMR (CDCl₃) δ 0.50- 0.60 (m, 6 H), 0.87 - 0.98 (m, 15 H), 0.98 - 1.91 (m, 17 H), 2.27 (m, 2 H), 4.01 (m, 1 H), 7.23 - 7.36 (m, 5 H); ¹³C NMR (CDCl₃) δ 4.9, 7.0, 13.5, 17.7, 18.4, 20.8, 23.0, 25.2, 27.2, 34.7, 35.1, 35.2, 37.9, 40.7, 42.1, 52.2, 53.0, 56.3, 69.4, 126.1, 126.7, 128.6, 144.1, 211.3; HRMS calcd for C₃₂H₅₆O₂SiNa⁺ [M + Na⁺]: 523.3942. Found: 523.3955. [0151] Preparation of C,D ring ketone 50. In a flame-dried, 25-mL, round-bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed 48 (0.028 g, 0.057 mmol) in THF (2 mL). To this solution was added tetrabutylammonium fluoride (0.11 mL, 1 M in THF, 0.11 mmol) and the resulting mixture was stirred at room temperature for 16 hours. Then, it was quenched with H₂O (5 mL) and extracted with EtOAc (3x 5 mL). The combined organic layers were washed with H₂O (Ix 5 mL) and brine (Ix 5 mL), dried (MgSO₄), and concentrated. Flash chromatography

(EtOAc:hexane, 20:80) afforded 0.019 g (91%) of the alcohol as an oil; [α£⁵ = +31.5 (c 0.48, CHCl₃); IR (neat cm^"1) 3518, 2942, 1701. ¹H NMR (CDCl₃) δ 0.83 (d, J= 6.4 Hz, 3 H), 0.88 (s, 3 H), 0.97 - 1.61 (m, 16 H), 1.50 (s, 6 H), 1.67 - 1.98 (m, 4 H), 2.21 (m, 2 H), 4.06 (m, 1 H), 7.23 - 7.36 (m, 5 H); ¹³C NMR (CDCl₃) δ 13.5, 17.4, 18.4, 20.8, 22.5, 25.1, 25.2, 27.0, 33.5, 35.1, 35.2, 37.9, 40.3, 41.8, 52.2, 52.5, 56.3, 69.4, 126.1, 126.7, 128.6, 213.3; HRMS calcd for C₂₅H₃₈O₂Na⁺ [M + Na⁺]: 393.2764. Found: 393.2774.

[0152] In a flame-dried, 25-mL, round-bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed the alcohol (0.019 g, 0.052 mmol) in dichloromethane (2 mL). To this solution was added pyridinium dichromate (0.039 g, 0.10 mmol) and the resulting mixture was stirred at room temperature for 20 hours. Then, it was filtered through a pad of Celite and concentrated. Flash chromatography (EtOAc:hexane, 20:80) afforded 0.015 g (80%) of 50 as an oil; [ajf = +3.8 (c 0.43, CHCl₃); IR (neat) 2962, 1708 cm^"1; ¹H NMR (CDCl₃) δ 0.58 (s, 3 H), 0.88 (d, J= 6.4 Hz, 3 H), 1.50 (s, 6 H), 1.12 - 2.26 (m, 18 H), 2.43 (m, 1 H), 7.23 - 7.36 (m, 5 H); ¹³C NMR (CDCl₃) δ 12.4, 18.5, 19.0, 20.6, 24.0, 25.1, 25.2, 27.3, 35.1, 35.2, 37.7, 38.9, 40.9, 49.9, 52.2, 56.3, 61.9, 126.1, 126.8, 128.9, 212.1, 213.2; HRMS calcd for C₂₅H₃₆O₂Na⁺ [M + Na⁺]: 391.2607. Found: 391.2604.

[0153] Preparation of 25(O)-Me₂Ph 8. In a flame-dried, 15-mL, round-bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed the phosphine oxide (-)-31 (0.020 g, 0.034 mmol) in THF (1 mL). To this solution was added n-BuLi (0.026 mL, 1.5 M in hexane, 0.038 mmol) at -78 ⁰C and the resulting red colored mixture was stirred at this temperature for 15 minutes, hi a separate, flame-dried, 15-mL, pear-shaped, single-necked flask fitted with a rubber septum and an argon balloon was placed the C,D-ring ketone 50 (0.0080 g, 0.022 mmol) in THF (1 mL). This solution was cooled -78 ⁰C, and transferred to the flask containing the phosphine oxide anion at -78 ⁰C via cannula. After stirring the reaction mixture at this temperature for 3 hours, it was quenched with pH 7 buffer solution (1 mL) and extracted with EtOAc (5x 3 mL). The combined organic phases were washed with H₂O (Ix 5 mL) and brine (Ix 5 mL), dried (MgSO₄), and concentrated. Flash chromatography (EtOAc:hexane, 10:90) afforded 0.0090 g (56%) of the coupled product as a yellow oil.

[0154] The coupled product (0.0090 g, 0.012 mmol) was dissolved in THF (3 mL), and to this solution was added tetrabuylammonium fluroide (0.3 mL, 1.0 M in THF, 0.3 mmol). After stirring the mixture at room temperature for 17 hours in dark, it was concentrated and purified by flash chromatography (EtOAc :hexane, 90:10) to afford 0.0050 g (83%) of 8 as a yellow oil. This was further purified by chiral HPLC (OD semipreparation column; 2-

Proρanol:Hexane = 10:90; flow rate = 2.5 mL/min; P = 0.24 kpsi) to afford 0.0019 g (31% overall yield from the C,D-ring ketone 50) of 8 (t_R= 20.5 min) as a colorless oil; [α]" = +20.4 (c 0.1, CHCl₃); IR (neat cm^'1) 3366, 2944, 1707. ¹H NMR (CDCl₃) δ 0.49 (s, 3 H), 0.84 (d, J= 6.3 Hz, 3 H), 1.51 (s, 6 H), 1.01 - 2.45 (m, 23 H), 2.60 (m, 1 H), 2.81 (m, 1 H), 4.22 (m, 1 H), 4.43 (m, 1 H), 5.01 (s, 1 H), 5.33 (s, 1 H), 6.00 (d, J= 11.2 Hz, 1 H), 6.39 (d, J= 11.3 Hz, 1 H), 7.23 - 7.36 (m, 5 H); ¹³C NMR (CDCl₃) δ 12.0, 18.7, 20.8, 22.2, 23.6, 25.2, 27.5, 29.1, 35.3, 35.9, 37.9, 40.4, 42.8, 45.3, 45.9, 52.2, 56.2, 56.3, 66.9, 70.8, 111.8, 117.0, 125.0, 126.8, 128.6, 132.8, 143.3, 144.1, 147.6, 213.4; HRMS Calcd for C₃₄H₄₈O₃Na⁺ [M + Na⁺]: 527.3496. Found: 527.3479; UV (MeOH) λmax 264 run (e 15,565).

EXAMPLE 9 25(OVAdamantyl 9 (Figure 23)

[0155] Preparation of TES ether 52. In a flame-dried, 25-mL, round-bottomed, single- necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed diisoproylamine (0.070 mL, 0.50 mmol) in THF (1 mL). The solution was cooled to -78 ⁰C followed by the addition of n-BuLi (0.31 mL, 1.6 M in hexane, 0.50 mmol), and was stirred for 10 minutes. To this was added a solution of 1-adamantyl methyl ketone (0.089 g, 0.50 mmol) in THF (1 mL), and the resulting mixture was stirred for 30 min followed by the addition of hexamethylphosphoramide (0.20 mL). The reaction mixture was stirred for an additional 15 min, and to this was added a solution of iodide 22 (0.030 g, 0.067 mol) in THF (1 mL). The mixture was slowly warmed to room temperature and stirred for 16 hours. Then, it was quenched with H₂O (3 niL) and extracted with EtOAc (3x 3 mL). The combined organic phases were washed with H₂O (Ix 3 mL) and brine (Ix 3 mL), dried (MgSO₄), and concentrated. Flash chromatography (EtOAc :hexane, 5:95) afforded 0.030 g

(89%) of 52 as an oil: [α£⁵ +38.2 (c 1.0, CHCl₃); IR (neat, cm^"1) 2906, 1700; ¹HNMR 5 (CDCl₃, 400 MHz).δ 4.01 (m, 1 H), 2.40 (m, 2 H), 2.04-0.98 (m, 32 H), 0.98-0.87 (m, 15 H), 0.60-0.50 (m, 6 H); ¹³CNMR (CDCl₃, 100 MHz) δ 216.0, 69.4, 53.1, 46.3, 42.1,, 40.8, 38.2, 36.6, 36.5, 35.5, 35.1, 34.6, 28.0, 27.3, 23.0, 20.1, 18.6, 17.7, 13.5, 6.9, 4.9; HRMS calcd for C₃₂H₅₆O₂SiNa⁺ [M + Na⁺] 523.3942 found 523.3955.

[0156] B. Preparation of C,D ring ketone 54. In a flame-dried, 25-mL, round-bottomed, 0 single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed 52 (0.028 g, 0.056 mmol) in THF (2 mL). To this solution was added tetrabutylammonium fluoride (0.11 mL, 1 M in THF, 0.11 mmol) and the resulting mixture was stirred at room temperature for 16 hours. Then, it was quenched with H₂O (5 mL) and extracted with EtOAc (3x 5 mL). The combined organic layers were washed with H₂O (Ix 5 5 mL) and brine (Ix 5 mL), dried (MgSO₄), and concentrated. Flash chromatography

(EtOAc.hexane, 20:80) afforded 0.022 g (99%) of the alcohol as an oil; [α]" +27.4 (c 1.0, CHCl₃); ¹HNMR (CDCl₃, 400 MHz) δ 4.01 (m, 1 H), 2.40 (m, 2 H), 2.04-0.98 (m, 32 H), 0.98-0.87 (m, 6 H), ¹³CNMR (CDCl₃, 100 MHz) δ 216.0, 69.4, 56.3, 52.6, 46.3, 41.8, 40.3, 38.2, 36.6, 36.4, 35.4, 35.1, 33.6, 28.0, 27.1, 22.5, 20.0, 18.5, 17.4, 13.5; IR (neat, cm^"1) 0 3508, 2906, 1696; HRMS calcd for C₂₆H₄₂O₂SiNa⁺ [M + Na⁺] 409.3077 found 409.3054. hi a flame-dried, 25-mL, round-bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed the alcohol (0.020 g, 0.052 mmol) in dichloromethane (2 mL). To this solution was added pyridinium dichromate (0.039 g, 0.10 mmol) and the resulting mixture was stirred at room temperature for 20 hours. Then, it was 5 filtered through a pad of Celite and concentrated. Flash chromatography (EtOAchexane,

20:80) afforded 0.016 g (80%) of 54 as an oil: [αg = +7.1 (c 1.0, CHCl₃); ¹H NMR (CDCl₃, 400 MHz) δ 2.41-0.98 (m, 34 H), 0.93 (d, J = 6.3 Hz, 3 H), 0.61 (s, 3 H); ¹³C NMR (CDCl₃, 100 MHz) δ 215.8, 212.2, 62.0, 56.3, 50.0, 46.3, 41.0, 38.9, 38.2, 36.6, 36.3, 35.4, 28.0, 27.6, 24.1, 19.9, 19.0, 18.7, 12.5; IR (neat, cm^"1) 2904, 1713; HRMS calcd for 0 C₂₆H₄₀O₂SiNa⁺ [M+Na] 407.2920 found 407.2932.

[0157] C. Preparation of 25(O)-Adamantyl 9. hi a flame-dried, 15 -mL, round- bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed the phosphine oxide (-)-31 (0.050 g, 0.086 mmol) in THF (1 niL). To this solution was added n-BuLi (0.050 mL, 1.6 M in hexane, 0.080 mmol) at -78 °C and the resulting red colored mixture was stirred at this temperature for 15 minutes. In a separate, flame-dried, 15-mL, pear-shaped, single-necked flask fitted with a rubber septum and an argon balloon was placed the enantiomerically pure C,D-ring ketone (+)-54 (0.016 g, 0.042 mmol) in THF (1 mL). This solution was cooled -78 ⁰C, and transferred to the flask containing the phosphine oxide anion at -78 °C via cannula. After stirring the reaction mixture at this temperature for 3 hours, it was quenched with pH 7 buffer solution (1 mL) and extracted with EtOAc (5x 3 mL). The combined organic phases were washed with H₂O (Ix 5 mL) and brine (1x 5 mL), dried (MgSO₄), and concentrated. Flash chromatography (EtOAc :hexane, 10:90) afforded 0.0060 g (19%) of the coupled product as a yellow oil. The coupled product (0.0060 g, 0.0080 mmol) was dissolved in THF (3 mL), and to this solution was added tetrabuylammonium fluroide (0.3 mL, 1.0 M in THF, 0.3 mmol). After stirring the mixture at room temperature for 17 h in dark, it was concentrated and purified by flash chromatography (EtOAc :hexane, 90:10) to afford 0.0058 g (72%) of 9 as a yellow oil. This was further purified by chiral HPLC (OD semipreparation column; 2- Propanol:Hexane = 7:93; flow rate = 2.5 mL/niin; P = 0.27 kpsi) to afford 0.0021 g

(25%) of 7 (t_Λ = 28.1 min) as a colorless oil; [αg +10.2 (c 0.1, CHCl₃); ¹H NMR (CDCl₃, 400 MHz) δ 6.39 (d, J= 11.2 Hz, 1 H), 6.00 (d, J= 11.3 Hz, 1 H), 5.33 (s, 1 H), 5.01 (s, 1 H), 4.42 (m, 1 H), 4.21 (m, 1 H), 2.81 (m, 1 H), 2.60 (m, 1 H), 2.48 (m, 3 H), 2.00-1.01 (m, 35 H), 0.93 (d, J= 6.4 Hz, 3 H), 0.54 (s, 3 H); ¹³C NMR (CDCl₃, 100 MHz) δ 215.9, 147.6, 143.3, 132.8, 125.0, 117.0, 111.8, 70.8, 66.9, 56.3, 56.2, 46.3, 45.9, 45.3, 42.8, 40.4, 38.2, 36.6, 36.4, 36.0, 35.6, 29.7, 29.1, 28.0, 27.6, 23.6, 22.2, 20.1, 18.8, 12.0; IR (neat, cm ¹) 3445, 1638;

HRMS calcd for C₃₅H₅₂O₃Na⁺ [M+Na] 543.3801 found 543.3822; UY (MeOH) λ_max 263 nm (ε 6,827).

EXAMPLE 10 Hydroxy ketone analog 10 (Figure 24)

[0158] A. Preparation of Hydroxy diketone 60. To a solution of enolate formed by treatment of 3-(trimethylsilyloxy)-3-methyl-2-butanone (100.20 mg, 1.2 eq) in THF (3 mL) at -78 ⁰C with LDA (generated at -78 ⁰C in situ) by 359.27 μL of n-BuLi (1.6 M in hexanes), 1.2 eq. and 80.56 μL of diisopropylamine (1.2 eq) was added the aldehyde 37 (162.2 mg, 0.48 mmol) in THF (2 mL). The reaction was slowly warmed to room temperature and allowed to stir overnight. Water was added, the reaction was extracted with EtOAc, dried over MgSO₄, purified by column chromatography (Hex. only) to give 130.4 mg (45%) of enone 56 (E : Z = 20 : 1 confirmed by ¹H NMR) and 98.3 mg of alcohol 57 (40%) as colorless oils. A solution of enone 56 (90 mg, 0.18 mmol) in anhydrous benzene (15 mL) was hydrogenated for 12h in the presence of 10% Pd/C (60 mg) until the absence of 56 was indicated by TLC. The reaction was filtered through a pad of celite, washed with hexanes, concentrated, and purified by column chromatography (2% EtOAc/hexanes) to afford 67.51 mg (75%) of the desired product as a colorless oil. The saturated bis-silylated ketone 58 (47.96 mg, 0.097 mmol) in THF (2 mL) was treated with TBAF (0.39 mL, 0.39 mmol, 1.0 M in THF) and allowed to stir at room temperature for 16 hours. A general workup and flash silica gel chromatography (25% EtOAc/hexanes) provided 30.0 mg (90%) of the diol as a colorless oil: IR (neat, cm^"1) 3436, 2931, 2860, 1707, 1460, 1372, 1360, 1266, 1166, 1067, 984, 961, 937, 884, 861; HRMS calcd for C₁₉H₃₄O₃Na⁺ [M + Na⁺] 333.2400 found 333.2377. To a solution of diol (30.0 mg, 0.097mmol) in CH₂Cl₂ (10 mL) was added 109.06 mg of PDC (0.29 mmol) and 109 mg of celite at room temperature. The reaction mixture was allowed to stir overnight at room temperature. The mixture was passed through 2 cm pad of silica gel, washed with EtOAc, concentrated, and purified by column chromatography (40% EtOAc/hexanes) to give 23.4 mg (79%) of ketone 60 as a colorless oil: [αg +6.18 (c 1.15, CHCl₃); ¹H NMR (CDCl₃, 400 MHz) δ 3.80 (brs, IH), 2.53-2.48 (m, 2 H), 2.43 (dd, J= 11.2, 7.6 Hz, IH), 2.28-2.19 (m, 2H), 2.12-2.08 (m, IH), 2.01-1.96 (m, IH), 1.93-1.83 (m, 2H), 1.77-1.66 (m, 3H), 1.60-1.41 (m, 5H), 1.47-1.41 (m, IH), 1.35 (s, 6H), 1.12-1.05 (m, IH), 0.96 (d, J= 6.0 Hz, 3H), 0.62 (s, 3H) ; HRMS calcd for C₁₉H₃₂O₃Na⁺ [M + Na⁺] 331.2243 found 333.2219.

[0159] B. Preparation of TMS-C₅D ring diketone 62. To a solution of diketone 60 (23.0 mg, 0.075 mmol) in anhydrous CH₂Cl₂ (5 mL) was added 21.88 μL of 1- (trimethylsilyl)-imidazole (0.15 mol) at room temperature. The reaction was allowed to stir overnight at room temperature. The reaction was evaporated in vacuo, the residue was purified by column chromatography (30% EtOAc/hexanes) to give 25.0 mg (88%) of the desired product as a colorless oil: [αjf +4.72 (c 1.20, CHCl₃); ¹H NMR (CDCl₃, 400 MHz) δ 2.63-2.55 (m, 2H), 2.43 (dd, J= 11.6, 7.2 Hz₅ IH), 2.29-2.17 (m, 2H), 2.13-2.07 (m, IH), 2.02-1.96 (m, IH), 1.95-1.85 (m, 2H), 1.75-1.66 (m, 2H), 1.63-1.58 (m, IH), 1.57-1.49 (m, 2H), 1.46-1.35 (m, 4H), 1.31 (s, 6H), 1.11-1.03 (m, IH), 0.96 (d, J= 6.4 Hz, 3H), 0.62 (s, 3H), 0.14 (s, 9H); ¹³C NMR (CDCl₃, 100 MHz) δ 215.57, 212.02, 80.03, 61.91, 56.28, 56.26, 49.84, 40.89, 38.88, 36.31, 35.35, 35.33, 27.39, 27.20, 27.18, 23.99, 19.97, 18.98, 18.64, 12.37, 2.22, IR (neat, cm^'1) 2942, 2872, 1707, 1460, 1372, 1360, 1307, 1249, 1196, 1037, 884, 837, 755, 685, 549; HRMS calcd for C₂₂H₄₀O₃SiNa⁺ [M + Na⁺] 403.2639 found 403.2649.

[0160] C. Preparation of Hydroxy ketone analog 10. To a solution of enantiomerically pure (-)-31 (30.2 mg, 0.051 mmol) in anhydrous THF (2 mL) was added 32.38 μL of nBuLi (0.051 mmol, 1.6 M in hexanes) at -78 ⁰C. After 15 min stirring, a precooled (-78 ⁰C) solution of enantiomerically pure C₅D ring ketone (+)-62 (13.15 mg, 0.034 mmol) in THF (1 mL) was added dropwise. The resulting reddish solution was allowed to stir for 4 hours at -78 ⁰C. The reaction was quenched by adding 1.0 mL of pH 7 buffer, then warmed to room temperature, extracted with EtOAc (20 mL x 2), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (10% EtOAc/hexanes) to afford 17.68 mg (69%) of the coupled product as a colorless oil.

[0161] The coupled product (17.65 mg, 0.023 mmol) was dissolved in 5 mL of anhydrous THF, and to this solution was added 0.19 mL (0.19 mmol) of TBAF (1.0 M in THF). The reaction was run in darkness overnight, then extracted with EtOAc (30 mL x 2), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (5% MeOH/CH₂Cl₂) to give 9.68 mg (92%) of the desired product 10 as a colorless oil: [αg +17.8 (c 0.75, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.37 (d, J= 11.2 Hz, IH)₅ 6.00 (d, J= 11.2 Hz, IH), 5.32 (s, IH), 4.99 (s, IH), 4.44-4.40 (m, IH), 4.24-4.21 (m, IH)₅ 3.84 (brs, IH), 2.82 (d, J= 12.4 Hz, IH), 3.61-2.56 (m, IH), 2.52-2.47 (m, 2H)₅ 2.30 (dd, J= 13.6, 6.4 Hz₅ IH), 2.01-1.98 (m, 2H), 1.95-1.82 (m, 3H), 1.79-1.65 (m, 6H)₅ 1.53-1.42 (m, 5H), 1.37 (s, 6H)₅ 1.33-1.25 (m, 2H), 1.10-1.01 (m, IH)₅ 0.94 (d, J= 6.4 Hz₅ 3H)₅ 0.53 (S₅ 3H); ¹³C NMR (100 MHz₅ CDCl₃) δ 214.60, 147.65, 143.09, 132.92, 124.97, 117.05, 111.75, 76.10, 70.82, 66.85, 56.31, 56.23, 45.89, 45.26, 42.87, 40.45, 35.98, 35.9I₅ 35.46, 29.06, 27.59, 26.52, 26.49, 23.55, 22.24, 20.29, 18.72, 11.98; IR (neat, crn ¹) 3378, 2942, 2872, 1701, 1642, 1465, 1443, 1372, 1190, 1055, 955, 908, 795, 726, 650, 591; HRMS rn/z (M + Na⁺) calcd 467.3131 for C₂₈H₄₄O₄Na⁺, found 467.3146.

EXAMPLE Il HBJ-24fMD25((J)TB 11 (Figure 25)

[0162] A. Preparation of 24-Amide silyl ether 64. To a solution of trimethylamide (51 mg, 0.50 mmol) in THF (5 mL) was added sodium hydride (12 mg, 0.50 mmol). After stirring for 1 hour at room temperature, a solution of the iodide 22 (45 mg, 0.10 mmol) in THF (2 mL) was added via cannula at room temperature. And then, the mixture was heated to reflux for 20 hr. The reaction mixture was slowly cooled to room temperature and then concentrated. The residue was subjected to column chromatography with EtOAc/hexanes

(1/4) as eluent to afford 40 mg (95 %) of the amide 64 as a colorless oil. : [α]^⁵ +46.7 (c 1.0, CHCl₃); ¹HNMR (400 MHz, CDCl₃) δ 5.53 (br s, IH), 4.02 (m, IH), 3.31 (m, IH), 3.14 (m, IH), 1.93 (dm, J_d = 12.4 Hz, IH), 1.74-1.86 (m, 2H), 1.66 (m, IH), 1.51-1.62 (m, 2H), 1.29-1.45 (m, 4H), 1.16-1.25 (m, 3H), 1.18 (s, 9H), 0.99-1.13 (m, 2H), 0.94 (t, J= 8.0 Hz, 9H), 0.94 (d, J= 6.4 Hz, 3H), 0.89 (s, 3H), 0.54 (q, J= 8.0 Hz, 6H); ¹³C NMR (100 MHz, CDCl₃) δ 178.2, 69.3, 56.5, 53.0, 42.1, 40.7, 38.5, 37.0, 35.5, 34.5, 33.4, 27.6, 27.4, 22.9, 18.7, 17.6, 13.4, 6.9, 4.9; IR (neat, cm^"1) 3342, 2952, 2875, 1637, 1537, 1458, 1373, 1230, 1165, 1083, 1025, 725; HRMS: calcd for C₂₅H₄₉NO₂SiNa⁺ [M + Na⁺]: 446.3425, found 446.3439.

[0163] B. Preparation of 24- Amide C,D~ring ketone 66. To a solution of silyl ether 64 (38 mg, 0.090 mmol) in THF (3 mL) was added 0.35 mL (0.35 mmol) of a 1.0 M solution of TBAF in THF at room temperature, and then it was stirred overnight at room temperature. The reaction mixture was quenched with water (2 mL), extracted with EtOAc (20 mL x 3), washed with brine, dried over MgSO₄, concentrated. The residue was subjected to column chromatography with EtOAc/Hexanes (1/1) as eluent to give the desired alcohol as a colorless oil.

[0164] To a solution of the alcohol in CH₂Cl₂ (4 mL) was added 118 mg of oven-dried Celite and PDC (118 mg, 0.32 mmol) at room temperature. The mixture solution was stirred overnight and then passed through a 2 cm pad of flash silica gel and washed with EtOAc. The filtrate was concentrated and subjected to column chromatography with EtOAc/Hexanes (2/3) as eluent to give 25 mg (91 %) of the desired C,D-ring ketone 66 as a colorless oil: [αg +12.7 (c 1.0, CHCl₃); ¹HNMR (400 MHz, CDCl₃) δ 5.64 (br s, IH), 3.29 (m, IH), 3.13 (m, IH), 2.41 (dd, J= 11.6, 7.2 Hz, IH), 2.14-2.27 (m, 2H), 2.07 (ddd, J = 13.2, 4.0, 2.4 Hz, IH), 1.79-2.02 (m, 3H), 1.69 (m, IH), 1.38-1.60 (m, 5H), 1.21-1.31 (m, 2H), 1.15 (s, 9H), 0.97 (d, J= 6.0 Hz, 3H), 0.59 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 211.8, 178.2, 61.8, 56.2, 49.8, 40.8, 38.8, 38.5, 36.8, 35.5, 33.6, 27.5, 23.9, 18.9, 18.7, 12.3; IR (neat, cm^"1) 3352, 2958, 2873, 1712, 1640, 1532, 1479, 1378, 1308, 1230, 1214, 1056; HRMS: calcd for C₁₉H₃₃NO₂Na⁺ [M + Na⁺]: 330.2403, found 330.2415.

[0165] C. Preparation of 24(NH)25(O)TB 11. A solution of 47 mg (0.081 mmol) of enantiomeric pure phosphine oxide (-)-31 in 1.5 mL of anhydrous THF was cooled to -78 ⁰C and treated with 50 μL (0.081 mmol, 1.6 M in hexanes) of n-BuLi under argon atmosphere. The mixture turned deep reddish and was stirred for 15 min at -78 ⁰C. To the solution was added dropwise a precooled (-78 ⁰C) solution of 8 mg (0.026 mmol) of the C,D-ring ketone (+)-66 in 1.5 mL of anhydrous THF via cannula. The reaction kept going until the reddish orange color faded to yellow (about 5 hr). The reaction was quenched by adding 1.0 mL of pH 7 buffer at -78 ⁰C, then warmed to room temperature, extracted with EtOAc (20 mL x 3), washed with brine, dried over MgSO₄, concentrated. The residue was subjected to column chromatography with EtOAc/hexanes (1/4) as eluent to afford 9 mg (53 %) of the coupled product as a colorless oil.

[0166] The coupled product (9 mg, 0.013 mmol) was dissolved in 3 mL of anhydrous THF, and to the solution was added 0.13 mL (0.13 mmol) of a 1.0 M solution of TBAF in THF. The resulting mixture was stirred in darkness overnight at room temperature, then quenched with 1 mL of water. The solution was extracted with EtOAc (20 mL x 3), washed with brine, dried over MgSO₄, concentrated. The residue was subjected to column chromatography with EtOAc as eluent to give 5 mg (94 %) of the desired product 11 as a colorless oil.: [αg +20.1 (c 0.25, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.37 (d, J= 11.2 Hz, IH), 6.01 (d, J= 11.2 Hz, IH), 5.54 (br s, IH), 5.33 (s, IH), 5.00 (s, IH), 4.43 (m, IH), 4.23 (m, IH), 3.32 (m, IH), 3.17 (m, IH), 2.82 (dd, J= 12.0, 4.0 Hz, IH), 2.60 (dd, J= 13.6, 3.2 Hz, IH), 2.31 (dd, J= 13.2, 6.4 Hz, IH), 1.89-2.04 (m, 5H), 1.42-1.71 (m, 7H), 1.14-1.35 (m, 6H), 1.19 (s, 9H), 0.98 (d, J= 6.4 Hz, 3H), 0.54 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 178.3, 147.6, 143.0, 132.9, 124.9, 117.1, 111.8, 70.8, 66.8, 56.27, 56.25, 45.9, 45.3, 42.8, 40.4, 38.6, 37.1, 35.7, 34.3, 29.0, 27.8, 27.6, 23.5, 22.2, 18.9, 11.9; IR (neat, cm^{" !}) 3346, 2949, 2872, 1635, 1539, 1455, 1437, 1349, 1214, 1057, 754; HRMS: calcd for C₂₈H₄₅NO₃Na⁺ [M + Na⁺]: 466.3292, found 466.3312. The product 11 was further purified by HPLC (t_R = 23.4 min, Chiralcel OJ column, 10 % 2-propanol in Hexanes, 2.5 mL/min, 254 nm) for testing the biological activity.

EXAMPLE 12 24-(NMe)25(OVrB 12 fFigure 25)

[0167] A. Preparation of 24-(N-methyl)amide C, D-ring ketone 67. To a solution of N-methyltrimethylamide (65 mg, 0.56 mmol) in THF (5 mL) in sealed tube was added sodium hydride (14 mg, 0.56 mmol). After stirring for 1 hr at room temperature, a solution of the iodide 22 (50 mg, 0.11 mmol) in THF (2 mL) was added via cannula at room temperature. And then, the mixture was heated at 100 ⁰C for 20 hr. The reaction mixture was slowly cooled to room temperature and then concentrated. The residue was subjected to column chromatography with EtOAc/hexanes (1/6) as eluent to afford 13 mg (30 %) of the amide 65 as a colorless oil.

[0168] To a solution of silyl ether 65 (13 mg, 0.030 mmol) in THF (2 mL) was added 0.10 mL (0.10 mmol) of a 1.0 M solution of TBAF in THF at room temperature, and then it was stirred overnight at room temperature. The reaction mixture was quenched with water (2 mL), extracted with EtOAc (20 mL x 3), washed with brine, dried over MgSO₄, concentrated. The residue was subjected to column chromatography with EtOAc/Hexanes (1/1) as eluent to give the desired alcohol as a colorless oil.

[0169] To a solution of the alcohol in CH₂Cl₂ (3 mL) was added 37 mg of oven-dried Celite and PDC (37 mg, 0.10 mmol) at room temperature. The mixture solution was stirred overnight and then passed through a 2 cm pad of flash silica gel and washed with EtOAc. The filtrate was concentrated and subjected to column chromatography with EtOAc/Hexanes (1/2) as eluent to give 9 mg (94 %) of the desired C,D-ring ketone 67 as a colorless oil: [α]* +8.1 (c 0.50, CHCl₃); ¹HNMR (400 MHz, CDCl₃) δ 3.26-3.43 (m, 2H), 3.01 (s, 3H), 2.44 (dd, J= 11.6, 7.6 Hz, IH), 2.17-2.30 (m, 2H), 2.10 (ddd, J= 12.8, 4.4, 2.4 Hz, IH), 1.85-2.04 (m, 3H), 1.26-1.78 (m, 8H), 1.26 (s, 9H), 1.02 (d, J= 6.0 Hz, 3H), 0.63 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 211.9, 177.1, 61.9, 56.4, 49.9, 47.9, 40.9, 38.9, 38.7, 36.3, 33.9, 32.9, 28.3, 27.6, 24.0, 19.0, 18.9, 12.4; IR (neat, cm-¹) 2957, 2874, 1713, 1626, 1480, 1404, 1378, 1367, 1307, 1116, 1080.

[0170] B. Preparation of 24(NMe)25(O)TB 12. A solution of 35 mg (0.060 mmol) of enantiomeric pure phosphine oxide (-)-31 in 1.5 mL of anhydrous THF was cooled to -78 ⁰C and treated with 38 μL (0.060 mmol, 1.6 M in hexanes) of «-BuLi under argon atmosphere. The mixture turned deep reddish and was stirred for 15 min at -78 ⁰C. To the solution was added dropwise a precooled (-78 ⁰C) solution of 9 mg (0.028 mmol) of the C,D-ring ketone (+)-67 in 1.0 mL of anhydrous THF via cannula. The reaction kept going until the reddish orange color faded to yellow (about 4 hr). The reaction was quenched by adding 1.0 mL of pH 7 buffer at -78 ⁰C, then warmed to room temperature, extracted with EtOAc (20 mL x 3), washed with brine, dried over MgSO₄, concentrated. The residue was subjected to column chromatography with EtOAc/hexanes (1/6) as eluent to afford 6 mg (31 %) of the coupled product as a colorless oil.

[0171] The coupled product (6 mg, 0.0087 mmol) was dissolved in 2 mL of anhydrous THF, and to the solution was added 0.09 mL (0.090 mmol) of a 1.0 M solution of TBAF in THF. The resulting mixture was stirred in darkness overnight at room temperature, then quenched with 1 mL of water. The solution was extracted with EtOAc (20 mL x 3), washed with brine, dried over MgSO₄, concentrated. The residue was subjected to column chromatography with EtOAc as eluent to give 4 mg (95 %) of the desired product 12 as a colorless oil: [<xj? +62.2 (c 0.10, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.37 (d, J= 11.2 Hz, IH), 6.01 (d, J= 11.2 Hz, IH), 5.33 (s, IH), 5.00 (s, IH), 4.43 (m, IH), 4.23 (m, IH), 3.29-3.42 (m, 2H), 3.01 (s, 3H), 2.82 (dd, J= 12.0, 4.0 Hz, IH), 2.60 (dd, J= 13.2, 3.2 Hz, IH), 2.31 (dd, J= 13.2, 6.4 Hz, IH), 1.89-2.01 (m, 5H), 1.22-1.71 (m, HH), 1.27 (s, 9H), 0.99 (d, J= 6.4 Hz, 3H), 0.54 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 177.1, 147.6, 143.0, 133.0, 124.9, 117.1, 111.8, 70.8, 66.8, 56.30, 56.27, 48.0, 45.9, 45.2, 42.8, 40.4, 38.7, 36.3, 34.5, 33.1, 29.0, 28.3, 27.7, 23.5, 22.2, 19.1, 11.9; IR (neat, cm ¹) 3388, 2946, 2872, 1608, 1408, 1364, 1261, 1214, 1114, 1056, 753.

[0172] The product 12 was further purified by HPLC (t_R = 28.9 min, Chiralcel OD column, 10 % 2-propanol in Hexanes, 2.5 mL/min, 254 nm) for testing the biological activity. EXAMPLE 13

24(NMe)-25(O)-26(Me-c-Hex) 13 (Figure 26)

[0173] A. Preparation of 24(N-methyl) amide C,D ring ketone 69. Sodium hydride (20 mg, 0.5 mmol) was added to a solution of methyl c-hexanyl amide (70 mg, 0.50 mmol) in DMF (1.5 mL) at room temperature. After 1 hour, 22 was transferred into the flask via cannula. The reaction was quenched with NH₄Cl after 16 hours. The mixture was extracted with ether for 4 times, washed with brine solution and water 5 times, dried, removed solvent and separated on radial chromatography (1:9, 1:4, 3:7 EtOAc/hexanes) to afford 22 mg (71%) of 68 as a colorless oil. MeLi (62 μL, 0.1 mmol) was added to a solution of 68 (22mg, 0.048 mmol) in 1.5 mL THF at -78 ⁰C for 10 minutes. MeI (14.2 mg, 0.1 mmol) was then introduced one portion. The reaction was quenched with NH₄Cl after an overnight at room temperature. Normal workup gives a crude product (16 mg) which was dissolved in 2 mL of THF and stirred at room temperature overnight with TBAF (1.0M, 0.2 mL). The reaction was quenched with water, extracted with ether for 4 times, washed with brine, dried with MgSO_4, removed solvent and purified on radial chromatography (2:3 EtOAc/hexanes) to afford 16 mg (91% for 2 steps) 63 as a colorless oil. 2 mL OfCH₂Cl₂ was added to a flask with 16 mg of 63, PDC (83 mg, 0.22 mmol) and 83 mg celite at room temperature. The mixture introduced onto a column (1 :2 EtOAc/hexanes as the eluent) to give 69, 11 mg (0.031 mmol, 69%) as a light yellow oil: [α Jf +68.84 (c 0.55, CH₃Cl); ¹H NMR (400 MHz, CDCl₃) δ 3.37 (m, 2H), 3.02 (s, 3H), 2.45 (dd, J= 12.0, 7.2 Hz, IH), 2.31-1.25 (m, 24H), 1.21 (s, 3H), 1.02 (d, J= 6.0 Hz, 3H), 0.63 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 213.0, 177.5, 63.0, 57.5, 50.9, 49.1, 44.1, 41.2, 40.0, 38.2, 37.6, 35.0, 34.1, 30.8, 28.7, 27.1, 25.6, 25.1, 24.3, 20.1, 20.0, 13.5; IR (neat, cm^"1) 1714, 1625; HRMS calcd for (M + Na) C₂₃H₃₉NO₂Na⁺ 384.2873, found 384.2869.

[0174] B. Preparation of 24(NMe)-25(O)-26(Me-c-Hex) 13. R-BuLi (1.6 M, 0.035 mmol) was added to a solution of 20.0 mg (0.034 mmol) of enantiomeric pure (-)-31 in 1 mL THF at -78⁰C. The mixture showed deep reddish and was stirred for 10 min at the same temperature. Precooled C₅D ring ketone (6 mg, 0.017mmol) in 0.5 mL THF was transferred via cannula to the A-ring solution. The reaction was allowed to proceed in the dark until the reddish color disappeared in 5 hours at the same temperature. The reaction was quenched with pH=7 buffer at - 78°C, extracted with ether for 4 times, washed with brine, dried over F

MgSO₄, purified on radial chromatography (1:9, 1:4, 3:7 EtOAc/hexanes) to give 5.5 mg of the coupled product as a colorless oil. 5.5 mg of the coupled compound in 1 mL THF was treated with TBAF (IM, 0.03 mL) at room temperature overnight. The mixture was quenched with pH=7 buffer, extracted with ether 4 times, washed with brine, dried over 5 MgSO₄ and purified on radial chromatography (70% EtOAc/hexanes) to give 13 3.3 mg as a white solid: [ag +17.44 (c 0.17, CH₃Cl); ¹H NMR (400 MHz, CDCl₃) δ 6.38 (d, J= 11.6 Hz, IH), 6.01 (d, J= 11.6 Hz, IH), 5.33 (s, IH), 5.00 (s, IH), 4.43 (m, IH), 4.23 (m, IH), 3.37 (m, 2H), 3.00 (s, 3H), 2.84-2.80 (m, IH), 2.61-2.58 (m, IH), 2.31 (dd, J= 14.0, 6.4 Hz, IH), 2.07-1.92 (m, 5H), 1.68-1.16 (m, 23H), 1.21 (s, 3H), 0.99 (d, J= 6.4 Hz, 3H), 0.54 (s, 10 3H); ¹³C NMR (100 MHz, CDCl₃) O 176.1, 147.6, 143.0, 132.9, 125.0, 117.1, 111.8, 70.8, 66.8, 56.3, 48.2, 45.9, 45.2, 43.0, 42.8, 40.4, 37.2, 34.6, 29.7, 29.0, 27.7, 26.0, 23.5, 23.2, 22.2, 19.0, 11.9; IR (neat, cm^"1) 3402, 1726, 1605; HRMS calcd for (M + Na⁺) C₃₂H₅₁NO₃Na⁺ 520.3761, found 520.3764.

15 EXAMPLE 14

Lactam analog 14 (Figure 27)

[0175] A. Preparation of TES-ether 70. To a solution of dimethylpyrrolidinone (37.30 mg, 0.33 mmol) in anhydrous DMF (5 mL) was added 13.19 mg of NaH (0.33 mmol) at 20 room temperature. After stirring for 1 hour at room temperature, a solution of iodide 22 (49.5 mg, 0.11 mmol) in DMF (2 mL) was added. The reaction mixture was allowed to stir overnight at room temperature. Water was added, the reaction mixture was extracted with EtOAc (3 x 20 mL), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (10% EtOAc/hexanes) to give 38.0 mg (79%) of the

25 desired lactam 70 as a colorless oil: [α£^s +30.6 (c 1.80, CHCl₃); ¹H NMR (400 MHz,

CDCl₃) δ 4.00 (d, J= 2.4 Hz, IH), 3.38-3.28 (m, IH), 3.27-3.13 (m, 3H), 1.92 (d, J= 12.4 Hz, IH), 1.82 (t, J = 6.8 Hz, 3H), 1.79-1.76 (m, IH), 1.66-1.58 (m, 2H), 1.57-1.50 (m, IH), 1.37-1.30 (m, 4H), 1.24-1.14 (m, 3H), 1.11 (s, 6H), 1.09-1.00 (m, 2H), 0.93 (t, J = 8.0 Hz, 9H), 0.91 (d, J= 6.4 Hz, 3H), 0.87 (s, 3H), 0.53 (q, J= 8.0 Hz, 6H); ¹³CNMR (100 MHz,

30 CDCl₃) δ 178.18, 69.31, 56.57, 53.00, 43.44, 42.15, 40.70, 40.59, 40.05, 34.56, 34.16, 33.18, 32.84, 27.34, 24.53, 24.46, 22.91, 18.50, 17.61, 13.36, 6.90, 4.89; IR (neat, cm^'1) 2942, 2861, 1684, 1490, 1460, 1425, 1372, 1278, 1231, 1161, 1084, 1014, 973, 943, 920, 902, 849, 802, 767, 720; HRMS m/z (M + Na⁺) calcd 458.3425 for C₂₆H₄₉NO₂SiNa⁺, found 458.3438.

[0176] B. Preparation of C₅D ring ketone 71. To a solution of silyl ether 70 (35.0 mg, 0.080 mmol) in THF (5 niL) was added 0.24 mL of TBAF (1.0 M in THF, 0.24 mmol) at room temperature. The reaction was allowed to stir overnight at room temperature. Water was added, the reaction mixture was extracted with EtOAc (3x 20 mL), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (50% EtOAc/hexanes) to give 34.0 mg (93%) of the alcohol as a colorless oil. To a mixture of the alcohol (24.0 mg, 0.075 mmol), 84.25 mg of PDC (0.22 mmol), and 85 mg of celite was added 10 mL OfCH₂Cl₂ at rt. The reaction was allowed to stir overnight at roomtemperature. The reaction mixture was passed through 2 cm pad of silica gel and washed with EtOAc. The filtrate was concentrated and purified by column chromatography (60% EtOAc/hexanes) to give 22.6 mg (95%) of the C₅D ring ketone 71 as a colorless oil:

[α£⁴ -3.30 (c 1.40, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 3.35 (dt, J= 13.6, 8.0 Hz, IH), 3.27-3.14 (m, 3H), 2.42 (dd, J= 11.6, 7.6 Hz, IH), 2.27-2.15 (m, 2H), 2.10-2.06 (m, IH), 2.01-1.86 (m, 3H), 1.82 (t, J= 6.8 Hz, 2H), 1.73-1.58 (m, 2H), 1.56-1.42 (m, 2H), 1.40-1.33 (m, IH), 1.30-1.15 (m, 3H), 1.09 (s, 6H), 1.00 (d, J= 6.4 Hz, 3H), 0.59 (s, 3H); IR (neat, cm^'1) 2954, 2860, 1707, 1678, 1560, 1542, 1501, 1454, 1431, 1378, 1360, 1284, 1225, 1172, 1055, 949, 843, 726, 579; HRMS m/z (M+Na) calcd 342.2403 for C₂₀H₃₃NO₂Na⁺, found 342.2408.

[0177] C. Preparation of Lactam analog 14. To a solution of enantiomerically pure (-)- 31 (30.0 mg, 0.051 mmol) in anhydrous THF (2 mL) was added 32.17 uL of ra-BuLi (0.051 mmol, 1.6 M in hexanes) at -78 °C. After 15 min stirring, a precooled (-78 ⁰C) solution of the enantiomerically pure C,D ring ketone (-)-71 (10.96 mg, 0.034 mmol) in THF (1 mL) was added dropwise. The resulting reddish solution was allowed to stir for 4 hours at -78 °C. The reaction was quenched by adding 1.0 mL of pH 7 buffer, then warmed to room temperature, extracted with EtOAc (20 mL x 2), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (30% EtOAc/hexanes) to afford 14.5 mg (62%) of the coupled product as a colorless oil. The coupled product (13.9 mg, 0.020 mmol) was dissolved in 5 mL of anhydrous THF, and to this solution was added 81.26 μL (0.081 mmol) of TBAF (1.0 M in THF). The reaction was run in darkness overnight, then extracted with EtOAc (30 mL x 2), washed with brine, dried over MgSO₄, concentrated in vacuo, and then purified by column chromatography (5% MeOH/CH₂Cl₂) to give 7.4 mg (80%) of the desired product 14 as a colorless oil: [α£⁴ +10.1 (c 0.30,

CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 6.37 (d, J= 11.2 Hz, IH), 6.00 (d, J= 11.2 Hz, IH), 5.32 (s, IH), 4.99 (s, IH), 4.44-4.42 (m, IH), 4.25-4.21 (m, IH), 3.40-3.33 (m, IH), 3.29- 3.15 (m, 3H), 8.32 (d, J= 12.4 Hz, IH), 2.60 (d, J= 13.6 Hz, IH), 2.33-2.28 (m, IH), 2.02- 1.97 (m, 2H), 1.95-1.88 (m, 2H)₅ 1.84 (t, J= 6.4 Hz, 2H), 1.72-1.60 (m, 4H), 1.53-1.42 (m, 3H), 1.37-1.18 (m, 6H), 1.13 (s, 6H), 0.99 (d, J= 6.4 Hz, 3H), 0.53 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 179.33, 147.70, 143.01, 133.01, 124.98, 117.13, 111.78, 70.86, 66.88, 56.39, 56.32, 45.97, 45.30, 43.58, 42.91, 40.67, 40.47, 40.21, 34.26, 34.10, 33.26, 29.08, 27.75, 24.62, 24.53, 23.57, 22.26, 18.82, 11.94; IR (neat, cm^"1) 3377, 2931, 2860, 1666, 1490, 1466, 1431, 1372, 1278, 1213, 1055, 955, 896, 750, 732; HRMS m/z (M+Na) calcd 478.3291 for C₂₉H₄₅NO₃Na⁺, found 478.3283.

EXAMPLE 15 24(NH>25(C0Ph 15 (Figure 28)

[0178] A. Preparation of C₅D ring ketone 73. In a flame-dried, 25-mL, round- bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed NaH (0.016 g, 0.67 mmol) in DMF (.1 niL). To this was added a solution of benzamide (0.081 g, 0.67 mmol) in DMF (1 mL). After stirring at room. temperature for 1 hour, the reaction mixture was added a solution of the iodide 22 (0.030 g, 0.067 mmol) in DMF (1 mL) was added via cannula. Then, the resulting mixture was heated at 80 °C for 16 hours, cooled to room temperature, quenched with H₂O, and extracted with diethyl ether (3x 5 mL). The combined organic phases were washed with H₂O (Ix 5 mL) and brine (Ix 5 mL), dried (MgSO₄), concentrated. Flash chromatography (EtOAc:hexane, 30:70) afforded 0.081 g (50%) of 72 which was dissolved in THF (3 mL). To this was added tetrabutylammonium fluoride (0.40 mL, 1.0 M in THF, 0.40 mmol), and the resulting mixture was stirred at room temperature for 18 hours. Then, it was quenched with H₂O (5 mL) and extracted EtOAc (3x 5 mL). The combined organic layers were washed with H₂O (Ix 5 mL) and brine (Ix 5 mL), dried (MgSO₄), and concentrated. Flash chromatography (EtOAc:hexane, 40:60) afforded 0.012 g (90%) of the alcohol which was dissolved in dichloromethane (2 mL). To this was added pyridinium dichromate (0.027 g, 0.073 mmol), and the resulting mixture was stirred at room temperature for 18 hours. Then, it was filtered through a pad of Celite and concentrated. Flash chromatography (EtOAc:hexane, 30:70) afforded 0.010 g (85%) of 73 as a colorless oil; [αj? +11.9 (c 0.50, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 0.65 (s, 3H), 1.05 (d, J = 6.1 Hz, 3 H), 1.11-2.36 (m, 15H), 2.46 (m, IH), 3.48 (m, IH), 3.52 (m, IH), 6.18 (bra, IH), 7.41-7.52 (m, 3H)₅ 7.75 (m, 2H); ¹³CNMR (100 MHz, CDCl₃) δ 12.5, 18.8, 19.0, 24.0, 27.6, 33.8, 35.6, 37.5, 38.9, 40.9, 49.9, 56.4, 61.9, 126.8, 128.6, 131.4, 134.7, 167.5, 211.9; IR (neat, cm^'1) 3336, 2955, 1710, 1639; HRMS calcd for C₂₁H₂₉NO₂Na [M + Na⁺] 350.2090 found 350.2100.

[0179] B. Preparation of 24(NH)-25(O)Ph 15. In a flame-dried, 15-mL, round- bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon was placed the phosphine oxide (±)-31 (0.066 g, 0.11 mniol) in THF (1 mL). To this solution was added n-BuLi (0.069 mL, 1.6 M in hexane, 0.11 mmol) at -78°C and the resulting red colored mixture was stirred at this temperature for 15 minutes, hi a separate, flame-dried, 15-mL, pear-shaped, single-necked flask fitted with a rubber septum and an argon balloon was placed the enantiomerically pure C,D-ring ketone (+)-73 (0.018 g, 0.060 mmol) in THF (1 mL). This solution was cooled -78 °C, and transferred to the flask containing the phosphine oxide anion at -78 °C via cannula. After stirring the reaction mixture at this temperature for 3 hours, it was quenched with pH 7 buffer solution (1 mL) and extracted with EtOAc (5x 3 mL). The combined organic phases were washed with H₂O (Ix 5 mL) and brine (Ix 5 mL), dried (MgSO₄), and concentrated. Flash chromatography (EtOAc:hexane, 20:80) afforded 0.0083 g (20%) of the coupled product as a yellow oil.

[0180] The coupled product (0.0083 g, 0.012 mmol) was dissolved in THF (3 ml), and to this solution was added tetrabutylammonium fluoride (0.3 mL, 1.0 M in THF, 0.3 mmol). After stirring the mixture at room temperature for 17 hours in the dark, it was concentrated and purified by flash chromatography (EtOAc :hexane, 90:10) to afford 0.0044 g (80%) of 15 as a mixture of two diastereomers. The mixture was purified by chiral HPLC (Whelk-O; 2-Proρanol:Hexane + 20:80; flow rate = 2.5 niL/min; t_R = 35.7 min) to afford 0.0028 g (10

% overall yield from the C,D-ring detone 73) of 15 as a colorless oil; [α]" +5.2(c 0.1, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 0.56 (s, 3H), 1.02 (d, J= 6.5 Hz, 3H), 1.01-2.80 (m, 21H), 3.41 (m, IH) , 3.59 (m,lH), 4.21 (m,lH), 4.42 (m, IH), 5.01 (s, IH), 6.00 (m, 2H), 6.39 (d, J= 11.2 Hz, IH), 7.41 (m, 3H) 7.80 (m, 2H); IR (neat, cm^'1) 3445, 2962, 1641: HRMS calcd for C₃₀H₄₁NO₃Na⁺ [M + Na⁺] 486.2979 found 486.2975; UV (MeOH) )w 266nm (e 15,405). EXAMPLE 16 SS-23-oxa-24NflH)25rOVrB (+) - 16 (Figure 29)

[0181] A. Preparation of SS-II-23-Oxa-24-N(H)25(O)TB-CD-Ring-8-OTES 75. A flame-dried 10 mL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon was charged with t-butyl hydroxamic acid (+)-74 (40 mg, 0.34 mmol) (Still, W. C, et al, J. Org. Chem., 43:1404 (1978)) and dissolved in 1.7mL freshly distilled DMF making a 0.5M solution. Then the flask was cooled down to 0 ⁰C ice bath. To this solution sodium hydride (11 mg, 0.41 mmol) was added and allowed to stir for 30 minutes before Iodide (+)-25 (30 mg, 0.069 mmol) was cannulated into the flask as a solution in DMF. This mixture was allowed to stir at 0 ⁰C for an additional 30 min, then gradually warmed up to room temperature for 5 hours. TLC showed the complete consumption of starting material. The reaction was quenched by addition of 15 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 30 mL). The combined extracts were washed with water (Ix 5 mL), and brine solution (Ix 5 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by flash column chromatography (25% ethyl acetate in hexanes) to afford 23 mg of O-alkylated product (+)-75 as an oil, in an

80% yield. [α£⁵ +64.97 (c 1.45, CHCl₃); IR (neat, cm^'1) 3228, 2952, 2880, 1645, 1500, 1471, 1457, 1366, 1235, 1167, 1090, 1014; ¹H NMR (400 MHz, CDCl₃) δ 8.30 (s, IH), 4.02-4.01 (d, IH, J= 2 Hz), 3.86-3.81 (dd, IH, J= 16, 3.2 Hz), 3.67-3.63 (m, IH), 1.93- 1.52 (m, 7H), 1.37-1.24 (m, 6H), 1.89 (s, 9H), 1.08-1.06 (d, 3H, J= 8.0 Hz), 0.95-0.90 (m, 12H), 0.56-0.50 (q, 6H, J= 16, 8.0 Hz); ¹³C NMR (100 MHz, CDCl₃) δ 175.9, 81.73, 69.20, 53.11, 52.74, 42.18, 40.51, 37.91, 35.37, 34.54, 27.21, 26.87, 22.99, 17.59, 17.29, 13.40, 6.91, 4.88; HRMS: calcd for C₂₄H₄₇NO₃SiNa⁺ [M + Na⁺]: 448.3217, found 448.3206.

[0182] B. Preparation of SS-II-23-Oxa-24-N(H)25(0)TB-CD-Ring-ketone (+)-76.

A flame-dried 10 mL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon was charged TES protected alcohol (+)-75 (23.0 μg, 0.054mmol) and dissolved in 2.70 mL freshly distilled THF. Then the flask was cooled to -78 ⁰C in an isopropanol/dry ice bath. To this solution 0.540 mL of Tetrabutylammonium fluoride (TBAF) (0.540 mmol, 1.0 M solution in THF) was added drop-wise over several minutes and the contents in the flask stirred at -78 ⁰C for an additional 30 minutes. The mixture was gradually warmed up to room temperature, and left stirring overnight. TLC showed the complete consumption of starting material. The reaction was quenched by addition of 5 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL), and brine solution (Ix 25 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product. This crude product was charged into an argon purged 10 mL recovery flask equipped with a magnetic stir bar, a septum and dissolved in 2.0 mL freshly distilled THF to give ca. 0.02 M solution. Then, to this solution were added PDC (35 mg, 0.096 mmol) and 26 mg of oven-dried Celite in one portion at room. The resulting mixture was allowed to stir at room temperature for 12 hours. TLC showed the complete consumption of starting material. The mixture was directly purified by column chromatography (50% ethyl acetate in hexanes) to afford 13 mg of ketone (+)-76 as an oil, in an 89 % yield, [α]^⁵ +17.12 (c 0.65, CHCl₃); IR (neat, cm^"1) 3248, 2960, 1710, 1645, 1484, 1458, 1381, 1370, 1231, 1056, 1005, 933; ¹H NMR (400 MHz, CDCl₃) δ 8.27 (s, IH), 3.83-3.80 (dd, IH, J= 12, 4 Hz), 3.76-3.73 (dd, IH, J= 8, 4 Hz), 2.46-1.99 (m, 7H), 1.74-1.56 (m, 6H), 1.20 (s, 9H), 1.17-1.15 (d, 3H, J= 8.0 Hz), 0.64 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 211.8, 176.1, 81.38, 61.61, 52.97, 49.85, 40.90, 38.72, 37.98, 35.61, 27.23, 27.09, 23.97, 19.13, 17.48, 12.39; HRMS: calcd for C₁₈H_3INO₃Na⁺ [M + Na⁺]: 332.2196, found 332.2192.

[0183] C. Preparation of SS-23-oxa-24N(H)25(0)TB (+)-16. Enantiomerically pure phosphine oxide (-)-31 and CD-ring ketone (+)-76, were separately azeotropically dried with anhydrous benzene (4x 5 mL) on a rotary evaporator and held under vacuum (ca. 0.1 mmHg) for at least 48 hours prior to use. A flame-dried 10 mL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon was charged with phosphine oxide (-)-31 (70.0 mg, 0.130 mmol) which was dissolved in 1.5 mL freshly distilled THF to give ca. 0.090 M solution. The flask was cooled to -78 ⁰C in an isopropanol/dry ice bath. To this solution π-BuLi (108 μL, 0.130 mmol, 1.20 M solution in hexanes) was added dropwise over several minutes during which time a deep red color developed and persisted. This mixture was allowed to stir at -78 ⁰C for an additional 10 minutes. Meanwhile, a flame- dried 10 mL recovery flask equipped with a magnetic stir bar and a septum along with an Ar balloon was charged with CD-ring ketone (+)-76 (13.0 mg, 0.042 mmol) and dissolved in 0.5 mL freshly distilled THF. The solution of CD-ring ketone was cooled to -78 ⁰C in an isopropanol/dry ice bath and was transferred dropwise into the flask containing the phoshine oxide anion at -78 ⁰C via cannula over several minutes. After the addition was complete, the deep red color persisted and the mixture was allowed to stir at -78 ⁰C for ca. 1.5 hours during which time it was visually checked. Upon observation of the light yellow color, the reaction was quenched at -78 ⁰C by addition of 5 mL of pH 7 buffer and allowed to come to room temperature. The mixture was then rinsed into a separatory funnel with ethyl acetate and extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL) and brine solution (Ix 25 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by column chromatography (25% ethyl acetate in hexanes) to afford 7 mgs of product, in a 35% yield. The protected analog was transferred in a flame dried 5 ml flask, equipped with a stir bar and purged with Argon. Furthermore, the product was dissolved in ImL THF and cooled down to -78 ⁰C where TBAF (0.100 mL, 0.103mmol) was added dropwise. The mixture was gradually warmed up to room temperature and left stirring overnight. The next day TLC showed consumption of starting material. The reaction was quenched by addition of 15 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 30 mL). The combined extracts were washed with water (Ix 5 mL), and brine solution (Ix 5 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by flash column chromatography (70% ethyl acetate in hexanes) to afford 2.50 mg of analog (+)-16, in a 17% yield for 2 steps, [αg +12.6 (c 0.125, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 8.18 (s, IH), 6.39-6.36 (d, IH, J= 10.8 Hz), 6.03-6.00 (d, IH, J- 11.6 Hz), 5.36 (s, IH), 5.36 (s, IH), 5.00 (m, IH), 4.43 (m, IH), 4.24 (m, IH), 3.86-3.84 (dd, IH, J=I 1.6, 3.2 Hz), 3.71- 3.65 (m, IH), 2.85-2.81 (dd, IH, J= 12.0, 4.4 Hz), 2.601-2.56 (m, IH), 2.34-2.29 (dd, IH, J = 13.2, 6.4 Hz), 2.05-1.89 (m, 6H), 1.77-1.44 (m, 10H), 1.21 (s, 9H), 1.14-1.12 (d, 3H, J= 8 Hz), 0.56 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 176.1, 147.6, 142.8, 133.0, 124.9, 117.1, 111.8, 81.70, 70.80, 66.84, 55.98, 52.85, 45.97, 45.24, 42.82, 40.22, 37.97, 36.15, 29.68, 27.24, 23.50, 22.67, 22.30, 17.54, 11.93; HRMS: calcd for C₂₇H₄₃NO₄Na⁺ [M + Na⁺]: 468.3084, found 468.3071; UV (MeOH) λ_max 265 nm (ε 17,631). EXAMPLE 17

SS-23-oxa-24NfMe)25(O)TB (+V17 (Figure 30)

[0184] A. Preparation of SS-II-23-Oxa-24N(Me)25(O)TB-CD-Ring-8-OTES (+)-17 A flame-dried 10 niL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon was charged with t-butyl hydroxamic acid (+)-77 (151 mg, 0.115 mmol) (Still, W. C; Kahn, N.; Mitra, A. J Org. Chem. 1978, 43, 14O4.)and dissolved in 2.3 mL freshly distilled DMF making a 0.500 M solution. Then the flask was cooled to 0 ⁰C in an ice bath. To this solution sodium hydride (44.0 mg, 0.173 mmol) was added and allowed to stir for 30 minutes before being cooled (0⁰C). Iodide (+)-25 (30.0 mg, 0.069 mmol) was cannulated into the flask as a 0.070M solution in DMF. This mixture was allowed to stir at 0 ⁰C for an additional 30 min, then it was gradually warmed up to room temperature for 5 hours. TLC showed the complete consumption of starting material. The reaction was quenched by addition of 15 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 30 mL). The combined extracts were washed with water (Ix 5 mL), and brine solution (Ix 5 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by flash column chromatography (25% ethyl acetate in hexanes) to afford 22.0 mg, 70.0%, of O-alkylated product (+)-78 as an oil. [α£⁵ +35.21 (c 2.25, CHCl₃); IR (neat, cm^"1) 2950, 2876, 1654, 1456, 1394, 1338, 1233, 1172, 1079, 1017, 733; ¹H NMR (400 MHz, CDCl₃) δ 4.04-4.03 (m, IH), 3.76-3.73 (dd, IH, J= 8, 3.2 Hz), 3.62-3.58 (t, IH, J= 8.4 Hz), 3.15 (s, 3H), 1.97-1.94 (m, IH), 1.80-1.66 (m, 4H), 1.65-1.55 (m, IH), 1.45-1.28 (m, 5H), 1.24 (s, 9H), 1.22-1.09 (m, 2H), 1.06-1.05 (d, 3H, J= 6.4 Hz), 0.970- 0.915 (m, 12H), 0.579-0.519 (m, 6H); ¹³C NMR (100 MHz, CDCl₃) δ 179.4, 77.52, 69.19, 53.48, 52.82, 42.35, 40.64, 35.54, 34.63, 34.56, 27.18, 27.08, 23.09, 19.13, 17. 63, 17.42, 13.52, 6.94, 4.94; HRMS: calcd for C₂₅H₄₉NO₃SiNa⁺ [M + Na⁺]: 462.3374, found 462.3383.

[0185] B. Preparation of SS-II-23-Oxa-24N(Me)25(O)TB-CD-Ring-ketone (+)-79.

A flame-dried 10 mL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon was charged TES protected alcohol (+)-78 (22.0 μg, 0.049 mmol) and dissolved in 5.0 mL freshly distilled THF. Then the flask was cooled to -78 ⁰C in an isopropanol/dry ice bath. To this solution 0.486 mL of TBAF (0.490 mmol, 1.00 M solution in THF) was added dropwise over several minutes and the contents in the flask stirred at -78 ⁰C for an additional 30 minutes. The mixture was gradually warmed up to room temperature, and left stirring overnight. TLC showed the complete consumption of starting material. The reaction was quenched by addition of 5 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL), and brine solution (Ix 25 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product. This crude product was charged into a argon purged 10 mL recovery flask equipped with a magnetic stir bar and a septum and dissolved in 3.0 mL freshly distilled CH₂Cl₂ to give ca. 0.020 M solution. Then, to this solution were added NMO (11.0 mg, 0.092 mmol), 4 A molecular sieves, and TPAP (0.800 mg, 0.002 mmol). The resulting mixture was allowed to stir at room temperature for about 1 hour. TLC showed the complete consumption of starting material. The mixture was passed through a pad of celite using a 50% ethyl acetate in hexanes eluent, and the eluent was concentrated. The crude product was directly purified by column chromatography (50% ethyl acetate in hexanes) to afford 10.0 mg of ketone (+)-79 in 99.0 % yield as a clear oil. [αξf +1.78 (c 0.125, CHCl₃); IR (neat, cm^"1) 2957, 2875, 1713, 1644, 1474, 1455, 1386, 1342, 1230, 1160, 999; ¹H NMR (400 MHz, CDCl₃) δ 3.78-3.75(dd, IH, J= 8.4, 3.6Hz), 3.69-3.65 (t, IH, J= 8.0 Hz), 3.18 (s, 3H), 2.48-2,44 (m, IH), 2.35-2.189 (m, 2H), 2.14-2.1 (m, IH), 2.07-1.71 (m, 4H), 1.66-1.52 (m, 4H), 1.43-1.32 (m, IH), 1.25 (s, 9H), 1.14-1.13 (d, 3H, J = 6.8 Hz), 0.679 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 211.37, 179.4, 77.05, 61.56, 53.29, 49.87, 40.89, 39.28, 38.83, 35.72, 34.75, 27.28, 27.17, 23.93, 19.24, 17. 57, 12.47; HRMS: calcd for C₁₈H₃₃NO₃Na⁺ [M + Na⁺]: 346.2353, found 346.2364.

[0186] C. Preparation of SS-23-oxa-24N(Me)25(0)TB (+)-l 7. Enantiomerically pure phosphine oxide (-)-31 (Daniewski, A. R., et al, J. Org, Chem., 67:1580 (2002)) and CD- ring ketone (+)-79, were separately azeotropically dried with anhydrous benzene (4x 5 mL) on a rotary evaporator and held under vacuum (ca. 0.1 mmHg) for at least 48 hours prior to use.

[0187] A flame-dried 10 mL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon was charged with phosphine oxide (-)-31 (30.0 mg, 0.052 mmol) and dissolved in 0.750 mL freshly distilled THF to give ca. 0.09 M solution. The flask was cooled to -78 ⁰C in an isopropanol/dry ice bath. To this solution n-BuLi (38.0 μL, 0.057 mmol, 1.50 M solution in hexanes) was added drop-wise over several minutes during which time a deep red color developed and persisted. This mixture was allowed to stir at -78 ⁰C for an additional 10 minutes. Meanwhile, a flame-dried 10 mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon was charged with CD-ring ketone (+)-79 (10.0 mg, 0.031 mmol) dissolved in 0.5 mL freshly distilled THF and cooled to -78 ⁰C in an isopropanol/dry ice bath. The solution of CD-ring ketone was transferred dropwise into the flask containing the phoshine oxide anion at -78 ⁰C via cannula over several minutes. After the addition was complete, the deep red color persisted and the mixture was allowed to stir at -78 ⁰C for ca. 1.5 hours during which time it was visually checked. Upon observation of a light yellow color, the reaction was quenched at -78 ⁰C by addition of 5 mL of pH 7 buffer and allowed to come to room temperature. The mixture was then rinsed into a separatory funnel with ethyl acetate and extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL) and brine solution (Ix 25 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by column chromatography (25% ethyl acetate in hexanes) to afford 13.0 mg of product, in a 62.0% yield. The O-silylated analog was transferred in a flame dried 5 ml flask, equipped with a stir bar and purged with Argon. Then, the product was dissolved in ImL THF and cooled to —78 ⁰C where TBAF (0.180 mL, 0.189 mmol) was added dropwise. The mixture was gradually warmed up to room temperature and left stirring overnight. The next day TLC showed consumption of starting material. The reaction was quenched by addition of 15 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 30 mL). The combined extracts were washed with water (Ix 5 mL), and brine solution (Ix 5 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by flash column chromatography (100% ethyl acetate) to afford 8.00 mg of analog (+)-17 as an oil (61% yield for 2 steps), [αjf +7.18 (c 0.200, CHCl₃); IR (neat, cm^'1) 3404, 2951, 2875, 1625,

1487, 1402, 1392, 1355, 1217, 1172, 1054, 984; ¹HNMR (400 MHz, CDCl₃) δ 6.39-6.36 (d, IH, J= 11.2 Hz), 6.04-6.01 (d, IH, J= 11.2 Hz), 5.33 (s, IH), 4.99 (s, IH), 4.45-4.42 (m, IH), 4.26-4.21 (m, IH), 3.77-3.74 (dd, IH, J= 8.0, 3.2 Hz), 3.66-3.59 (m, IH), 3.17 (s, 3H), 2.86-2.82 (dd, IH, J= 12.0, 3.6 Hz), 2.62-2.58 (m, IH), 2.36-2.31 (dd, IH, J= 14.0, 8.0 Hz), 2.06-1.28 (m, 16H), 1.25 (s, 9H), 1.11-1.09 (d, 3H, J= 6.4 Hz)₅ 0.59 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ 179.4, 147.7, 142.5, 133.3, 124.8, 117.3, 111.7, 77.21, 70.76, 66.85, 56.00, 53.39, 53.17, 46.03, 45.24, 42.89, 40.33, 39.28, 36.29, 34.70, 29.69, 29.01, 27.39, 27.18, 23.48, 22.39, 20.64, 17.64, 13.98, 12.01; HRMS: calcd for C₂₈H₄₅NO₄Na⁺ [M + Na⁺]: 482.3241, found 482.3257; UV (MeOH) )w 265 nm (e 15,895).

EXAMPLE 18

KSP-23-Oxa-25-(OV26-TB 18 (Figure 31)

[0188] A. Preparation of Lythgoe-Inhoffen Diol 80. An oven-dried 1 L three-necked flask was charged sequentially with NaHCO₃ (170 mg, 2.0 mmol), anhydrous MeOH (120 mL), anhydrous CH₂Cl₂ (350 mL), and Vitamin D₂ (11.42 g, 28.8 mmol). The solution was cooled to -78 ⁰C and treated with O₃ until a deep blue color developed and persisted (~2.5 h). The solution was subsequently flushed with O₂ for 30 min until the blue color faded. Solid sodium borohydride (9.5 g, 251 mmol) was added portion-wise over a period of 10 min at -78 ⁰C until complete disappearance of starting material was observed by TLC. The reaction mixture was warmed to 0 ⁰C and stirred for 3 hours. After being stirred for an addition 30 min at room temperature, the mixture was quenched with 1 N HCl, extracted the ethyl acetate (3 x 200 mL), dried over MgSO₄, filtered and concentrated. Purification by column chromatography afforded 4.11 g of 80 as a white solid in 65% yield. Spectroscopic data are identical to published reports, (Posner, G. H., et at, J. Org. Chem., 62:3299 (1997)).

[0189] B. Preparation of TIPS-Monoether (+)-82. A solution of diol 80 (85 mg, 0.40 mmol) in CH₂Cl₂ (3 mL) was cooled to 0 ⁰C and 2,6-lutidene (191 μL, 1.64 mmol) and TIPS triflate (237 μL, 0.88 mmol) were added. The solution was stirred at that temperature for 2 hours, then added to water and extracted with CH₂Cl₂ (3x 5 mL). The combined organic extracts were dried over MgSO₄, filtered, concentrated, and purified by column chromatography (100% hexanes) to afford 187 mg 81 as a colorless liquid in 90% yield.

[αg^'1 +34.3 (c 0.51, CHCl₃); IR (neat, cm^"1) 3735 (w), 2942 (s), 2890 (m), 2866 (s), 2360 (s), 2341 (m), 1541 (w), 1459 (m), 1339 (w), 1364 (w), 1245 (w), 1165 (w), 1097 (m), 1067 (w), 1036 (m), 1022 (m), 882 (m), 668 (s); ¹H NMR (CDCl₃, 400 MHz) δ 4.21 (q, IH, J= 2.8 Hz), 3.67 (dd, IH, J= 9.2, 3.2 Hz), 3.35 (dd, IH, J= 9.2, 7.2 Hz), 1.97 (dt, IH, J= 12.4, 2.8 Hz), 1.89-1.00 (m, 56 H), 0.95 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 69.96, 68.13, 53.26, 42.08, 40.73, 38.91, 34.62, 26.87, 23.43, 18.28, 18.23, 18.07, 17.80, 16.98, 13.95, 12.35, 12.04.

[0190] To a solution of 81 (171 mg, 0.326 mmol) at 0 ⁰C, was added a solution of 1 M TBAF in THF (360 μL) and the mixture was stirred for 14 hours. The mixture was quenched with water and extracted with CH₂Cl₂ (3x 5 mL). The combined organic extracts were dried over MgSO₄, filtered, concentrated, and purified by column chromatography to afford 110 mg of 82 as a colorless oil in 92% yield, [α£^8>1 + 37.6; IR (neat, cm^"1) 3355 (br),

2942 (s), 2866 (s), 2723 (w), 2360 (w), 1652 (w), 1463 (m), 136 (m), 1262 (m), 1216 (w), 1165 (m), 1092 (s), 1068 (m), 1024 (s), 949 (m), 922 (w)₅. 904 (w), 882 (m), 846 (w), 802 (w), 766 (m), 674 (s); ¹H NMR (CDCl₃, 400 MHz) δ 4.20 (q, IH, J= 2.8 Hz), 3.63 (dd, IH, J= 10.8, 3.2 Hz), 3.37 (dd, IH₅ J= 10.4, 6.8 Hz), 1.96 (m, IH), 1.90-1.65 (m, 4H), 1.54 (brs, IH), 1.46-1.01 (m, 32H) 0.96 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 69.87, 68.00, 53.24, 53.03, 42.09, 40.68, 38.29, 34.56, 26.81, 23.33, 18.27, 18.22, 17.77, 16.69, 13.93, 12.63; HRMS: calcd for C₂₂H₄₃O₂Si⁺ [M-H]⁺: 367.3032, found: 367.3058.

[0191] C. Preparation of (+)-85-8-OH-23-Oxa-25-Keto-26-TB. To ice-cooled hexanes-washed KH (120 mg of a 30% suspension in mineral oil, 0.9 mmol) was added a solution of 82 in THF (2.5 mL). The mixture was stirred at room temperature for 45 min until a light yellow color developed. Tosylate 83 (240 mg, 0.9 mmol) and Bu₄NI (6 mg, 0.02 mmol) were added to the reaction flask and the mixture was stirred at room temperature for 14 hours. The reaction was quenched by addition of water and extracted with CH₂Cl₂ (3x 5 mL). The combined organic extracts were dried over MgSO₄, filtered, concentrated, and purified by column chromatography to afford 100 mg of 84 as a colorless oil in 72% yield, [αg ¹ + 38.7 (c 0.28, CHCl₃); IR (neat, cm^"1) 2943 (s), 2866 (s), 1463 (m), 1361 (w), 1166 (m), 1098 (s), 1024 (m), 905 (w), 882 (m), 666 (s); ¹HNMR (CDCl₃, 400 MHz) δ 5.06 (m, IH), 4.98 (m, IH), 4.20 (m, IH), 3.96 (m, 2H), 3.39 (dd, IH, J= 8.8, 3.6 Hz), 3.12 (dd, IH₅ J= 8.8, 8 Hz), 1.97 (m, IH), 1.90-1.64 (m, 5H), 1.43-1.03 (m, 40H), 0.97 (s, 3H). ¹³C NMR (CDCl₃, 100 MHz) δ 154.05, 108.45, 75.77, 71.42, 69.94, 53.67, 53.24, 42.18, 40.69, 36.49, 34.79, 34.61, 29.44, 26.93, 23.39, 18.28, 18.24, 17.79, 17.58, 13.91, 12.65.

[0192] Through a solution of 84 (70 mg, 0.17 mmol) in CH₂Cl₂ (15 mL) in a three-necked flask at -78 ⁰C was bubbled O₃ until a deep blue color developed and persisted (approximately 20 min). The solution was subsequently flushed with O₂ for 15 min until the blue color faded. The colorless solution was cooled to 0 ⁰C and dimethyl sulfide (151 μL, 2.06 mmol) was added. The solution was allowed to warm slowly to room temperature and stir overnight. The solution was concentrated and subjected to column chromatography (10% ethyl acetate in hexanes) to afford 34 mg of crude 8-OTIPS-23(OXO)-25-KETO-TB. To a solution of the crude material in CH₃CN was added HF (49%, 300 μL, 7.3 mmol) and the solution was stirred at room temperature 14 hours. The mixture was quenched with a saturated solution OfNaHCO₃ and extracted with ethyl acetate (3x 10 mL). The combined organic extracts were rinsed with brine, dried over MgSO₄, filtered, concentrated, and purified by column chromatography (10-25% ethyl acetate in hexanes) to afford 10 mg of 85 as a colorless oil in 20% yield for the 3 steps. [α£" + 28.7 (c 0.15, CHCl₃); IR (neat, cm⁴) 3510 (br), 2934 (s), 2871 (s), 1720 (s), 1477 (m), 1368 (m), 1262 (w), 1158 (m), 1056 (m), 992 (m), 944 (m), 889 (w), 780 (w), 666 (w); ¹H NMR (CDCl₃, 400 MHz) δ 4.27 (m, IH), 4.07 (m, IH), 3.39 (dd, IH, J= 8.8, 3.6 Hz), 3. 21 (dd, IH, J= 8.8, 7.2 Hz), 1.99 (m, IH), 1.88-1.20 (m, 14H), 1.16 (s, 9H), 1.05 (d, 3H, J= 6.4 Hz), 0.95 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 212.12, 76.85, 72.17, 69.29, 53.33, 52.26, 42.87, 41.94, 40.21, 36.40, 33.60, 26.72, 26.28, 22.60, 17.40, 17.31, 13.55; HRMS: calcd for C₁₉H₃₄O₃Na⁺ [M + Na⁺]: 333.2400, found 333.2394.

[0193] D. Preparation of 8-Keto-23-Oxa-25-Keto-26-TB (+)-86

A solution of alcohol 85 (12.5 mg, 0.04 mmol) in CH₂Cl₂ (2 mL) was cannulated into a reaction vessel equipped with PDC (40 mg, 0.11 mmol) and oven dried Celite (40 mg). After stirring overnight the mixture was filtered, the filtrate concentrated in vacuo, and purified by column chromatography (10-25% ethyl acetate in hexanes) to afford 12 mg of

86 as a colorless oil in 95% yield, [αg ⁸ + 3.4 (c 0.675, CHCl₃); IR (neat, cm^'1) 3402 (w), 2960 (s), 2975 (s), 1715 (s), 1478 (m), 1426 (w), 1368 (m), 1308 (w), 1240 (m), 1220 (m), 1179 (w), 1150 (m), 1056 (m), 1001 (m), 942 (w), 837 (w), 720 (w); ¹H NMR (CDCl₃, 400 MHz) δ 4.28 (m, IH)₅ 4.07 (m, IH), 3.38 (dd, IH, J= 8.8, 3.6 Hz), 3. 27 (dd, IH, J= 8.8, 6.8 Hz), 2.45 (dd, IH, J= 11.6, 7.6 Hz), 1.88-1.20 (m, HH), 1.16 (s, 9H), 1.15 (d, 3H, J= 6.4 Hz), 0.65 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 212.08, 211.91, 76.54, 72.03, 61.66, 53.24, 49.90, 42.85, 40.94, 38.77, 36.60, 27.06, 26.26, 24.01, 19.16, 17.52. 12.48; HRMS: calcd for C₁₉H₃₂O₃Na⁺ [M + Na⁺]: 331.2244, found 331.2245. [0194] E. Preparation of 23-Oxa-25-(O)-26-TB (+)-l 8. Enantiomerically pure phosphine oxide (-)-31 (Daniewski, A. R., et at, J. Org. Chem., 67:1580 (2002)) and CD- ring ketone (+)-86, were separately azeotropically dried with anhydrous benzene (4x 5 mL) on a rotary evaporator and held under vacuum (ca. 0.1 mm Hg) for at least 48 hours prior to use.

[0195] A flame-dried 10 mL recovery flask equipped with a magnetic stir bar and a septum with an Ar ballon was charged with phosphine oxide (-)-31 (60 mg, 0.10 mmol) which was dissolved in 2.0 mL freshly distilled THF. The flask was cooled to - 78 ⁰C. To this solution n-BuLi (67 μL, 0.10 mmol, 1.60 M solution in hexanes) was added drop-wise over several minutes during which time a deep red color developed and persisted. This mixture was allowed to stir at - 78 ⁰C for an additional 10 minutes. Meanwhile, a flame- dried 10 mL flask containing CD-ring ketone 86 (12.0 mg, 0.04 mmol) was dissolved in 0.75 mL of freshly distilled THF and cooled to - 78 ⁰C. The solution of CD-ring ketone was transferred dropwise into the flask containing the phosphine oxide anion at — 78 ⁰C via cannula over several minutes. After the addition was complete, the deep red color persisted and the mixture was allowed to stir at - 78 ⁰C for 6 hours. Upon observation of a light yellow color, the reaction was quenched with 5 mL of pH 7 buffer and allowed to warm to room temperature. The mixture was extracted with ethyl acetate (3x 15 mL). The combined organic extracts were washed with water and brine, dried over MgSO₄, and filtered. The filtrate was concentrated in vacuo to give crude product that was purified by column chromatography (5-50% ethyl acetate in hexanes) affording 13.0 mg of product in a 50% yield. The protected analog was dissolved in CH₃CN (2 mL) and HF (49%, 15 μ) was added. After stirring 1 hour the reaction was quenched with a saturated solution of NaHCO₃ and extracted with ethyl acetate (3x 10 mL), dried over MgSO₄, and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by column chromatography (50-75% ethyl acetate in hexanes with 1% NEt₃) to afford 6 mg of analog

(+)-18 as an oil (40% yield for 2 steps), [αf⁴ + 11.2 (c 0.275, CHCl₃); IR (neat, cm^'1) 3422 (s), 1718 (m), 177 (w), 1654 (m), 1636 (m), 1419 (w), 1260 (w), 1056 (m), 754 (w); ¹H NMR (CDCl₃, 400 MHz) δ 6.38 (d, IH, J= 11.2 Hz), 6.02 (d, IH, J= 11.2 Hz), 5.33 (m, IH), 4.99 (m, IH), 4.29 (d, 2H, J= 2.4 Hz), 4.23 (m, IH), 3.42 (dd, IH, J= 8.8, 3.2 Hz), 3.21 (dd, IH, J= 8.8, 7.6 Hz), 2.82 (m, IH), 2.59 (m, IH), 2.31 (m, IH), 2.05-1.08 (m, 16H), 1.16 (s, 9H), 1.09 (d, 3H, J= 6.8 Hz), 0.58 (s, 3H); ¹³C NMR (CDCl₃, 100 MHz) δ 212.18, 147.63, 143.02, 132.96, 124.94, 117.06, 111.76, 76.85, 72.13, 70.80, 66.84, 56.03, 53.15, 46.00, 45.24, 42.85, 40.28, 3721, 29.06, 28.91, 27.21, 26.28, 23.53, 22.36, 17.56, 12.02; HRMS: calcd for C₂₈H₄₄O₄Na⁺ [M + Na⁺]: 467.3132, found 467.3162. UV (MeOH)

EXAMPLE 19

SS-25(O)26NfOMe)H 19 fFigure 32)

[0196] A. Preparation of SS-II-25(O)26N(OMe)H-CD-Ring-8-OTES (+)-89. In a flame-dried 25 -mL, two-necked, round-bottomed flask equipped with a magnetic stirring bar, a septum, a reflux condesor, and an argon balloon was placed activated zinc (0.086 g, 1.32 mmol), methyl acrylate (0.140 mL, 1.54 mmol), NiCl₂6H₂0 (0.040 g, 0.170 mmol) in pyridine (3mL). The mixture was heated at 65 ⁰C for 1 hour, and by then, the color of the reaction mixture become reddish brown. The reaction mixture was cooled to 0 ⁰C and added a solution of a pre-cooled (to O⁰C) iodide (+)-25 (0.050 g, 0.110 mmol). Then, it was warmed to room temperature and stirred for three hours. The mixture was diluted with EtOAc (5 mL) and filtered through a pad of Celite. The filtrate was washed with 5% HCl (2x 5 mL) and extracted with EtOAc (3x 5 mL). The combined extracts were washed with water (Ix 5 mL) and brine (Ix 5 mL), dried over NaSO₄, and concentrated. The crude product was purified via flash chromatography using 10% ethyl acetate in hexanes to afford 0.038 g of ester 88, as a oil, in an 88.0 % yield.

[0197] For the preparation of the 25(O)26N(OMe)H-CD-Ring-8-OTES 89, (Craig, D., et ah, Tetrahedron, 55:15025 (1999)) the purified ester 88 was transferred in a 10 ml pear shaped flasked and placed under vacuum for 24 hours. Li a flame dried 25 mL round bottom flask equipped with a magnetic stir bar, a septum and an argon balloon was placed well dried methoxylamine hydrochloride (0.007 g, 0.076 mmol) and dissolved in 10 mL in toluene to give ca. 0.08M solution. This mixture is cooled down to 0 ⁰C where the trimethylalumina (0.042 mL, 0.085 mmol) is added slowly dropwise. This proceeds to stir at 0 ⁰C for five minutes, then the reaction warms up to room temperature and it is left stirring for 1.5 hours before ester 88 (0.030 g, 0.076 mmol) is added via a cannula as a solution in benzene to give a ca. 0.08M solution. This was allowed to stir for 5 hours. TLC showed complete consumption of starting material. The reaction is quenched with water (Ix 10 mL), neutralized with 1 M HCl, and extracted with ethyl acetate (3x 30 mL). The extracted were combined, dried over NaSO₄, filtered and concentrated. The concentrate gave the crude product which was purified by flash column chromatography, which was eluted with 50% ethyl acetate to afford 21.0 mg of ester 89 as a colorless oil, in a 70.0% yield. Data for 89: [α£⁵ +34.39 (c 0.80, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 8.26 (s, IH), 4.02-4.01 (d, IH, J= 2.4Hz), 3.76 (s, 3H), 1.95-1.91 (m, IH), 1.86-1.04 (m, 18H), 0.96-0.89 (m, 15H), 0.57-0.51 (m, 6H); ¹³C NMR (100 MHz, CD₃OD) δ 171.42, 69.50, 62.92, 56.59, 52.97, 41.93, 40.67, 34.96, 34.86, 34.32, 32.73, 26.92, 22.81, 17.63, 17.33, 12.81, 5.93, 4.48; IR (neat, cm^'1) 3177, 2944, 2875, 1657, 1455, 1368, 1229, 1166, 1078, 1015, 971; HRMS: calcd for C₂₃H₄₅NO₃SiNa⁺ [M + Na⁺]: 434.3061, found 435.3051.

[0198] B. Preparation of SS-II-25-(0)26N(OMe)H-CD-Ring-ketone (-)-90. A flame- dried 10 mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon TES protected alcohol 89 (0.021 g, 0.049mmol) was charged and dissolved in 3.0 mL freshly distilled THF. Then the flask was cooled to -78 ⁰C in an isopropanol/dry ice bath. To this solution 0.485 mL of TBAF (0.485 mmol, 1.00 M solution in THF) was added dropwise over several minutes and the contents in the flask stirred at -78 ⁰C for an additional 30 minutes. The mixture was gradually warmed up to room temperature, and left stirring overnight. TLC showed the complete consumption of starting material. The reaction was quenched by addition of 5 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL), and brine solution (Ix 25 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product. This crude product was purified using flash chromatography 70% ethyl acetate in hexanes to afford 10 mg of deprotected alcohol in a 71% yield. The purified alcohol was charged into an argon purged 10 mL recovery flask equipped with a magnetic stir bar, a septum and dissolved in 2.00 mL freshly distilled THF to give ca. 0.02 M solution. Then, to this solution were added PDC (0.026 g, 0.071 mmol) and 16 mg of oven-dried Celite in one portion at room. The resulting mixture was allowed to stir at room temperature for about 12 hours. TLC showed the complete consumption of starting material. The mixture was directly purified by column chromatography eluted with 100% ethyl acetate afforded 9.00 mg of ketone (-)-90 as an oil, in a 89.0 % yield. Data for (-)-

90: [α]" -10.45 (c 0.355, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 8.22 (s, IH), 3.77 (s, 3H), 2.46-2.41 (m, IH), 2.33-2.16 (m, 3H), 2.12-1.97 (m, 3H), 1.95-1.82 (m, 3H), 1.77- 1.43 (m, 10H), 1.34-1.28 (m, 3H), 0.98-0.96 (d, 3H, J= 8Hz), 0.63 (s, 3H); IR (neat, cm⁴) 3210, 2956, 1711, 1652, 1456, 1378, 1070; HRMS: calcd for C₁₇H₂₉NO₃Na⁺ [M + Na⁺]: 318.2039, found 318.2034.

[0199] C. Preparation of SS-25(O)26N(OMe)H 19. Enantiomerically pure phosphine oxide (-)-31 and CD-ring ketone (-)-90, were separately azeotropically dried with anhydrous benzene (4x 5 mL) on a rotary evaporator and held under vacuum (ca. 0.1 mmHg) for at least 48 hours prior to use.

[0200] A flame-dried 10 mL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon was charged with phosphine oxide (-)-31 (59.0 mg, 0.100 mmol) and dissolved in 0.750 mL freshly distilled THF to give ca. 0.13 M solution. The flask was cooled to -78 ⁰C in an isopropanol/dry ice bath. To this solution n-BuLi (71.0 μL, 0.110 mmol, 1.60 M solution in hexanes) was added drop-wise over several minutes during which time a deep red color developed and persisted. This mixture was allowed to stir at -78 ⁰C for an additional 10 minutes. Meanwhile, a flame-dried 10 mL recovery flask equipped with a magnetic stir bar, a septum along with an Ar balloon was charged with CD-ring ketone (-)-90 (10.0 mg, 0.034 mmol) dissolved in 0.75 mL freshly distilled THF and cooled to -78 ⁰C in an isopropanol/dry ice bath. The solution of CD-ring ketone was transferred dropwise into the flask containing the phoshine oxide anion at -78 ⁰C via cannula over several minutes. After the addition was complete, the deep red color persisted and the mixture was allowed to stir at -78 ⁰C for ca. 2.0 hours during which time it was visually checked. Upon observation of a light yellow color, the reaction was quenched at -78 ⁰C by addition of 5 mL of pH 7 buffer and allowed to come to room temperature. The mixture was then rinsed into a separatory funnel with ethyl acetate and extracted with ethyl acetate (3x 25 mL). The combined extracts were washed with water (Ix 25 mL) and brine solution (Ix 25 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by column chromatography (50% ethyl acetate in hexanes) to afford 5.0 mg of product, in a 24.0% yield. The O-silylated analog was transferred in a flame dried 5 ml flask, equipped with a stir bar and purged with Argon. Then, the product was dissolved in ImL THF and cooled to -78 ⁰C where TBAF (0.040 mL, 0.075 mmol) was added dropwise. The mixture was gradually warmed up to room temperature and left stirring overnight. The next day TLC showed consumption of starting material. The reaction was quenched by addition of 15 mL distilled water and then rinsed into a separatory funnel with ethyl acetate. The mixture was extracted with ethyl acetate (3x 30 mL). The combined extracts were washed with water (Ix 5 mL), and brine solution (Ix 5 niL), dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuo to give the crude product that was purified by flash column chromatography (100% ethyl acetate) to afford 1.4 mg of analog 19 as an oil (11.2% yield for 2 steps). ¹H NMR (400 MHz, CDCl₃) δ 8.22 (s, IH), 6.39-6.36 (d, IH, J= 11.2 Hz), 6.02-6.00 (d, IH, J= 10.8 Hz), 5.33-5.32 (m, IH), 5.00 (s, IH), 4.44-4.43 (m, IH), 4.24- 4.22 (m, IH), 3.76 (s, IH), 2.84-2.81 (m, IH)₃ 2.62-2.59 (m, IH), 2.35-2.27 (m, IH), 2.04- 1.85 (m, 7H), 1.68-1.05 (m, 17H), 0.95-0.94 (d, 3H, J= 6.0 Hz), 0.54 (s, 3H); ¹³C NMR (100 MHz, CDCl₃) δ insufficient material for a carbon NMR; HRMS: calcd for C₂₆H₄₁NO₄Na⁺ [M + Na⁺]: 454.2927, found 454.2913; UV (MeOH) >w 265 nm (e 15,958).

EXAMPLE 20 24(O)26(O)TB 20 (Figure 33)

[0201] a. Preparation of ZZ-24(O)26(O)TB-CD-Ring-8-OTES(+)-92. In a flame- dried, round-bottomed, single-necked flask fitted with a magnetic stirring bar, a rubber septum, and an argon balloon, dione 91 (100 mg, 0.70 mmol) was added to a solution of sodium hydride (18.5 mg, 0.77 mmol) in 7mL of THF at 0 ⁰C. The reaction was allowed to stir for a half an hour upon which n-BuLi (1.6 M, 0.70 mmol) was added via a syringe, at 0 ⁰C. The reaction mixture was stirred for an additional 15 min, and to this was added a solution of iodide 25 (0.030 g, 0.067 mmol) in THF (1 mL), through the course of 20 minutes. The mixture was slowly warmed to room temperature and stirred for 60 hours. After 2.5 days the tic showed consumption of starting material, upon which the reaction was quenched with NH₄Cl solution, extracted with ether (4x 10 mL). The combined organic layers were washed with saturated NaCl solution, dried over MgSO₄ and concentrated. The crude product was separated on radial chromatography (1:9 EtOAc/hexanes) to afford 32 mg (+)-92 as a colorless oil. [α£⁴ +40.6 (c 1.6, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 0.52

- 0.58 (m, 6H), 0.54 (s, 3H), 0.91 - 0.96 (m, 9H), 0.91 (d, J= 6.4 Hz, 3H), 1.20 (s, 9H), 1.03 - 1.97 (m, 15H), 2.14 - 2.22 (m, IH), 2.32 - 2.40 (m, IH), 4.03 (d, J= 2.4 Hz, IH), 5.58 (s, IH); ¹³C NMR (100 MHz, CDCl₃) δ 4.98, 6.97, 13.6, 17.7, 18.4, 23.0, 26.0, 27.3, 27.3, 31.8, 34.7, 35.1, 35.9, 39.0, 40.8, 42.2, 53.1, 56.5, 69.4, 95.0, 196.6, 200.0; IR (neat, cm^"1) 2954, 2876, 1604, 1460, 1022; HRMS(ESI): calcd for (M + Na⁺) C₂₇H₅₀O₃SiNa⁺ 473.3421, found 473.3442.

[0202] b. Preparation of ZZ-C₉D Ring Tri-ketone (+)-93. A flame-dried 10 niL recovery flask equipped with a magnetic stir bar and a septum with an Ar balloon TES protected alcohol (+)-92 (30.0 mg, 0.066mmol) was charged and dissolved in 10 mL of MeCN. To this solution 0.540 mL of Hydrofluoric acid (HF) (48%, 6.6 mg, 0.33mmol) was added drop-wise over several minutes and the contents in the flask stirred at -78 ⁰C for an additional 5 minutes. TLC showed the complete consumption of starting material. The reaction was quenched by addition of 3 mL of saturated NaHCO₃ solution, extracted with ethyl ether (4x 1OmL). The organic layer was dried over MgSO₄ , filtered and concentrated. The product was purified via radial chromatography (3:7 EtOAc/hexanes), to afford the alcohol (20.1 mg, 91%) as a light yellow oil. The product was then charged into an argon purged 10 mL recovery flask equipped with a magnetic stir bar, a septum and dissolved in 5.0 mL freshly distilled DCM to give ca. 0.06 M solution. Then, to this solution, were added PDC (112.8 mg, 0.30 mmol) and 112.8 mg of oven-dried Celite in one portion at room. The resulting mixture was allowed to stir at room temperature for 6 hours. TLC showed the complete consumption of starting material. The mixture was directly purified by column chromatography. Flash column chromatography (10% ethyl acetate in hexanes) to afford C₅D ring triketone 93 (6.0 mg, 30% yield) as a colorless oil. [α]" 13.53 (c 0.30, CHCl₃); ¹H NMR (400 MHz, CDCl₃) δ 0.64 (s, 3H), 0.98 (d, J= 6.0 Hz, 3H), 1.17 (s, 9H), 1.32 - 2.47 (m, 17H), 5.59 (s, IH); ¹³C NMR (IOO MHZ, CDCl₃) δ 12.5, 18.5, 19.0, 24.0, 26.0, 27.3, 27.4, 31.6, 35.2, 35.8, 39.0, 40.9, 49.9, 56.4, 61.9, 95.0, 196.2, 200.0, 211.8; IR (neat, cm^"1) 2961, 2873, 1715, 1616, 1458; HRMS(ESI): calcd for (M + Na⁺) C₂₁H₃₄O₃Na⁺ 357.2400, found 357.2409.

[0203] c. Preparation of ZZ-24(O)26(O)TB (+)-20. In a flame-dried, 10-mL, round- bottomed, single-necked flask fitted with a magnetic stir bar, a rubber septum, and an argon balloon was placed the phosphine oxide (-)-31 (36 mg, 0.062mmol) in THF (1 mL). To this solution was added n-BuLi (0.034 mL, 1.6 M, 0.062 mmol) at -78 ⁰C and the resulting red colored mixture was stirred at this temperature for 20 minutes. In a separate, flame-dried, 15-mL, pear-shaped, single-necked flask fitted with a rubber septum and an argon balloon was placed the C,D-ring ketone 93 (6.0 mg, 0.018 mmol) in THF (1 mL). This solution was cooled -78 ⁰C, and transferred to the flask containing the phosphine oxide anion at -78 ⁰C via cannula. After stirring the reaction mixture at this temperature for 8.5 hours, it was quenched with pH 7 buffer solution (1 niL) and extracted with ethyl ether (5x 3 rnL). The combined organic phases were washed with H₂O (Ix 5 mL) and brine (Ix 5 mL), dried (MgSO₄), and concentrated. The crude product was purified on radial chromatography (1 :9, 1:4, 1:3 EtOAc/hexanes) to afford the coupled product (9.7 mg, 77%) as a white solid.

[0204] The coupled product (9.7 mg, 0.0139 mmol) was dissolved in CH₃CN (2 mL), and to this solution was added HF (48%, 0.067 mmol). After stirring the mixture at room temperature for 1 hour in dark, it was quenched with pH 7 buffer solution (1 mL) and extracted with ethyl ether (5x 3 mL). The combined organic phases were washed with H₂O (Ix 5 mL) and brine (Ix 5 mL), dried (MgSO₄), and concentrated. The crude product was purified by flash chromatography (EtOAc :hexane, 90:10) to afford 0.005O g (83%) of 20 as a yellow oil. This was further purified by chiral HPLC (OD semipreparation column; 2- Proρanol:Hexane = 10:90; flow rate = 2.5 mL/min; P = 0.24 kpsi) to afford 0.0019 g (31% αj_D = +20.4 (c 0.1, CHCl₃); IR (neat cm^"1) 3366, 2944, 1707. ¹H NMR (CDCl₃) δ 0.49 (s, 3 H), 0.84 (d, J= 6.3 Hz, 3 H), 1.51 (s, 6 H), 1.01 - 2.45 (m, 23 H), 2.60 (m, 1 H), 2.81 (m, 1 H), 4.22 (m, 1 H), 4.43 (m, 1 H), 5.01 (s, 1 H), 5.33 (s, 1 H)₃ 6.00 (d, J= 11.2 Hz, 1 H), 6.39 (d, J= 11.3 Hz, 1 H), 7.23 - 7.36 (m, 5 H); ¹³C NMR (CDCl₃) δ 12.0, 18.7, 20.8, 22.2, 23.6, 25.2, 27.5, 29.1, 35.3, 35.9, 37.9, 40.4, 42.8, 45.3, 45.9, 52.2, 56.2, 56.3, 66.9, 70.8, 111.8, 117.0, 125.0, 126.8, 128.6, 132.8, 143.3, 144.1, 147.6, 213.4; HRMS Calcd for C₃₄H₄₈O₃Na⁺ [M + Na⁺] : 527.3496. Found: 527.3479; UV (MeOH) X_m3x 264 nm (e 15,565).

[0205] The terms and expressions which have been employed herein are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding equivalents of the features shown and described, or portions thereof, it being recognized that various modifications are possible within the scope of the invention claimed. Moreover, any one or more features of any embodiment of the invention may be combined with any one or more other features of any other embodiment of the invention, without departing from the scope of the invention. For example, the features of the compounds of this invention are equally applicable to the methods of treating disease states described herein. AU publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

[0206] The References that appear throughout this application are all incorporated herein by reference.

Claims

WHAT IS CLAIMED IS: 1. An antiproliferative, low-calcemic compound having the Formula (II):

wherein L is selected from the following carbonyl side chains:

wherein n = 1, 2;

m = 0-2;

p - 3-6;

Z = CH₂, O, NR', S, SO, SO₂, O-N-R', N(R')-N(R'), -(C-O)-;

Y = O₅CH₂;

X ⁼ C, O, S, N; V = H, or V = V = CH₂;

R = isoalkyl, f-butyl, l-(Methyl)cycloalkyl (3-7), l-(OH)cycloalkyl (3-7), 1-adamantyl;

R = H, F, Me, CD₃, Et, /-Pr, allyl, CH₂OR, CMe₂Ph, R=R=cycloalkyl (3-7); and

R" = H, F, Me, CD₃, Et, i-Pr, allyl, CH₂OR, R=R=cycloalkyl (3-7).

2. The compound of claim 1, having the formula:

3. The compound of claim 1, having the formula:

4. The compound of claim 1, having the formula:

5. The compound of claim 1, having the formula:

6. The compound of claim 1, having the formula:

7. The compound of claim 1, having the formula:

8. The compound of claim 1, having the formula:

9. The compound of claim 1, having the formula:

10. The compound of claim 1, having the formula:

11. The compound of claim 1, having the formula:

12. The compound of claim 1, having the formula:

13. The compound of claim 1 , having the formula:

14. The compound of claim 1, having the formula:

15. The compound of claim 1, having the formula:

16. The compound of claim 1, having the formula:

17. The compound of claim 1, having the formula:

18. The compound of claim 1 , having the formula:

19. The compound of claim 1, having the formula:

20. The compound of claim 1, having the formula:

21. The compound of claim 1, having the formula:

22. The compound of claim 1, having the formula:

23. The compound of claim 1, having the formula:

24. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound having the Formula (II):

wherein L is selected from the following carbonyl side chains:

wherein n = 1, 2;

m = 0-2;

p = 3-6;

Z = CH₂, O, NR', S, SO, SO₂, 0-N-R¹, N(RO-N(R"), -(C=O)-;

Y = 0,CH₂;

X = C, O, S, N;

V = H, or V = V = CH₂;

R = isoalkyl, t-butyl, l-(Methyl)cycloalkyl (3-7), l-(OH)cycloalkyl (3-7), 1-adamantyl;

R' = H, F, Me, CD₃, Et, z-Pr, allyl, CH₂OR', CMe₂Ph, R=R'=cycloalkyl (3-7); and

R" = H, F, Me, CD₃, Et, /-Pr, allyl, CH₂OR', R'=R'=cycloalkyl (3-7).

25. The pharmaceutical composition of claim 24, having the formula:

26. The pharmaceutical composition of claim 24, having the formula:

27. The pharmaceutical composition of claim 24, having the formula:

28. The pharmaceutical composition of claim 24, having the formula:

29. The pharmaceutical composition of claim 24, having the formula:

30. The pharmaceutical composition of claim 24, having the formula:

31. The pharmaceutical composition of claim 24, having the formula:

32. The pharmaceutical composition of claim 24, having the formula:

33. The pharmaceutical composition of claim 24, having the formula:

34. The pharmaceutical composition of claim 24, having the formula:

35. The pharmaceutical composition of claim 24, having the formula:

36. The pharmaceutical composition of claim 24, having the formula:

37. The pharmaceutical composition of claim 24, having the formula:

38. The pharmaceutical composition of claim 24, having the formula:

39. The pharmaceutical composition of claim 24, having the formula:

40. The pharmaceutical composition of claim 24, having the formula:

41. The pharmaceutical composition of claim 24, having the formula:

42. The pharmaceutical composition of claim 24, having the formula:

43. The pharmaceutical composition of claim 24, having the formula:

44. The pharmaceutical composition of claim 24, having the formula:

45. The pharmaceutical composition of claim 24, having the formula:

46. The pharmaceutical composition of claim 24, having the formula:

47. A method for treating a disease resulting from the proliferation of cells, said method comprising; administering to a patient suffering from said disease an effective amount of a compound having the Formula (II):

wherein L is selected from the following carbonyl side chains:

wherein n = 1, 2;

m = 0-2;

p = 3-6;

Z = CH₂, O, NR¹, S, SO, SO₂, 0-N-R¹, N(R')-N(R'), -(C=O)-;

Y = O₅CH₂;

X = C₅ O₅ S₅ N;

V = H₅ Or V = V = CH₂;

R = isoalkyl, t-butyl, l-(Methyl)cycloalkyl (3-7), l-(OH)cycloalkyl (3-7), 1-adamantyl;

R = H₅ F₅ Me₅ CD₃, Et₅ /-Pr₅ allyl, CH₂OR', CMe₂Ph, R'=R'=cycloalkyl (3-7); and

R" = H, F, Me, CD₃, Et₅ /-Pr₅ allyl, CH₂OR¹, R'=R'=cycloalkyl (3-7).