SELF-ASSEMBLED TAXO-DIPERPENOID NANOSTRUCTURES

Abstract: Onium salts of taxo-diterpenoid-C<n>,2-O-aza-arenes form water soluble self-assembling nanostructures above a critical aggregation concentration. The onium salt of aza-arene includes a delocalized charge which renders derivatized taxo-diterpenoids amphoteric, enhances their solubility in water, and promotes self-assembly of water soluble nanostructures, e.g., helical fibular structures and spherical micellular structures. These same onium salts of taxo-diterpenoid-C<n>,2-O-aza-arenes are employed as water soluble prodrugs. For example, taxol-2'-methylpyridinium tosylate (MPT) is characterized by an elevated aqueous solubility, rapid activation by serum protein, good stability in most other aqueous solutions, formation of a protein: taxol intermediate and good retention within the circulatory system. The toxicity of the activated form is comparable or greater than underivatized taxol. Furthermore, taxol-2'-MPT can be synthesized by a simple one step reaction between taxol and 2-fluoro-1-MPT. The invention is applicable to taxol and taxol memetics having hydroxyls that are reactive with onium salts of 2-halogenated-aza-arenes. For example, taxol, C-2 substituted taxol and Taxotere each have reactive hydroxyls at the 2' and 7 positions. The invention is also applicable to a wide array of 2-halogenated-aza-arenes.

Description: <Desc/Clms Page number 1> SELF-ASSEMBLED TAXO-DITERPENOID NANOSTRUCTURES Description Technical Field: The invention relates to taxol prodrugs. More particularly, the invention relates to taxol and taxol memetics derivatized with onium salts of aza-arenes to form water soluble prodrugs which form stable self-assembled nanostructures in aqueous solvents having helical and micellular structures and which are converted to active form upon contact with serum protein. Background: Taxol, an antineoplastic agent originally isolated from Taxus brevifolia, is approved for usage in the treatment of ovarian cancer and is expected to see usage in breast, lung, and skin cancers as well. However, since Taxol possesses an extremely low water solubility, i.e., less than 1.5 x 10-6 M, it has been necessary to formulate Taxol in a mixture of CremaphorTM, a polyoxyethylated castor oil, and ethanol in order to achieve a therapeutic concentration. This formulation can induce a variety of significant side effects including hypersensitivity reactions. While premedication and slow administration of the drug can circumvent these problems in the clinic, the entire protocol is quite cumbersome and requires extensive close monitoring of patients. Although taxol's dramatic efficacy has driven clinical usage forward despite these problems, a water soluble form of taxol could completely obviate the need for this troublesome protocol. <Desc/Clms Page number 2> One approach to bypassing these formulation difficulties, previously attempted by several groups including our own, is the introduction of solubilizing functionality that normal metabolic pathways could remove in vivo. Compounds of this type, termed prodrugs, consist, in the case of taxol, primarily of ester derivatives at the 2' and 7 positions. Currently none of these protaxols have given success in the clinic. In each case, the prodrug is rapidly cleared from circulation by the kidneys. Taxol is only one of a class of taxoditerpenoids having bioactivity. Another preferred taxo-diterpenoid having clinically significant activity is Taxotere. Unfortunately, all known bioactive taxo-diterpenoids have a low aqueous solubility. What is needed is a stable prodrug which is activated by contact with serum and which is retained in circulation for a clinically significant period after administration. Summary: The invention is directed to water soluble selfassembling nanostructures of onium salts of taxo- EMI2.1 diterpenoid-C',2-0-aza-arenes and solutions thereof. The onium salt of aza-arene includes a delocalized charge which renders the molecule amphoteric and enhances its solubility in water. In a first embodiment, the invention herein teaches that water EMI2.2 soluble onium salts of taxo-diterpenoid-C",2-O-aza- arenes self-assemble in aqueous media to form water soluble helical nanostructure. In a second embodiment, the invention herein teaches that, with the aid of sonication, water soluble onium salts of EMI2.3 taxo-diterpenoid-C",2-O-aza-arenes self-assemble in <Desc/Clms Page number 3> aqueous media to form water soluble micellular nanostructure. The invention further teaches that these nanostructures are stable in aqueous media over a period of hours or days. It is also taught that, with the addition of serum protein, 2-0-aza-arene is displaced from the onium salt of taxo-diterpenoid- Cn,2-0-aza-arene and a soluble protein:taxoditerpenoid intermediate is formed. Furthermore, it is taught herein that the protein:taxo-diterpenoid intermediate dissociates over time to provide a bioactive taxo-diterpenoid. Preferred taxoditerpenoids include taxol, C-2 substituted taxol EMI3.1 analogs, and Taxotere. Taxo-diterpenoid-Cn,2-0-aza- arene may be produced in a one step synthesis by reacting onium salts of 2-halogenated aza-arenes with reactive hydroxyls on the taxo-diterpenoid. Reactive hydroxyls on taxol and Taxotere are located at C2' and C7. A preferred onium salt of 2-halogenated azaarene is 2-fluoro-1-methylpyridinium tosylate. Other employable onium salts of 2-halogenated aza-arenes are disclosed by T.Mukaiyama, Angewandte Chemie 1979, 18(18), 707-808, incorporated herein by reference. More particularly, the first embodiment of the invention is directed to water soluble self-assembled helical fibrous nanostructure having a diameter of 60-120 and a helical twist of 5-10 molecules per turn of an onium salt of a taxo-diterpenoid-2-O-azaarene wherein the solubility of the onium salt of the taxo-diterpenoid-2-O-aza-arene exceeds 10-3 moles per liter and the 2-0-aza-arene is labile to displacement by serum protein. The first embodiment of the invention is also directed to compositions comprising an aqueous solvent in combination with dissolved taxo-diterpenoid-2-O-aza-arenes capable of forming such water soluble self-assembled helical fibrous <Desc/Clms Page number 4> nanostructures above a critical aggregation concentration. A second embodiment of the invention is directed to water soluble self-assembled micellular nanostructures having a spherical shape with a diameter of 20-120 A and incorporating onium salts of taxo-diterpenoid-2-O-aza-arenes wherein the solubility of the onium salt of the taxo-diterpenoid- 2-0-aza-arene exceeds 10-3 moles per liter. The second embodiment of the invention is also directed to compositions comprising an aqueous solvent in combination with dissolved taxo-diterpenoid-2-0-azaarenes capable of forming water soluble selfassembled micellular nanostructures above a critical aggregation concentration upon sonication. The invention is also directed to a process for making self-assembled micellular nanostructures from an aqueous solution including an onium salt of a taxo- EMI4.1 diterpenoid=2-O-aza-arene. This process employs sonication at a concentration in excess of a critical aggregation concentration for the onium salt of the taxo-diterpenoid-2-O-aza-arene. In a preferred embodiment, the taxo-diterpenoid- 2-0-aza-arenes are represented by the following formula: EMI4.2 <Desc/Clms Page number 5> wherein Rx is Ph or tBuO; R10 is OAc or OH; Ry is a C-2 substituent defined below; and R2' and R7 are each selected from the group consisting of OH and an onium salt of a 2-0-aza-arene, with the proviso that at least one of R2' and R7 is the onium salt of the 2-0aza-arene. The onium salt of the 2-0-aza-arene can be represented by either of the following formulas for onium salt I or onium salt II: EMI5.1 wherein Z1 and Z2 are each either C or N; Z3 is S or 0; R1 is selected from the group consisting of C1-C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; R2 and R6 are each selected from the group consisting of H, CI-C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; if Z' is C, then R3is selected from the group consisting of H, Cj-C alkyl, allyl, arenxyl, propargyl, C1-C6 O-alkyl, OH, halogen, and fused aryl; if Z1 is N, then R3 is absent; R4 and R8 are each selected from the group consisting of H, CI-C6 alkyl, allyl, arenxyl, propargyl, C1-C6 O-alkyl, OH, halogen, and fused aryl; and if Z2 is C, then R5 is selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, Cj-C 0-alkyl, OH, halogen, and fused aryl; if Z2 is N, then R5 is absent; and S- is a counter ion. <Desc/Clms Page number 6> Preferred C-2 RY substitutions include benzyl and the following: EMI6.1 Brief Description of the Drawings: Figure 1 illustrates the kinetics of taxol release from taxol-2'-MPT (2) in various aqueous solutions at 25 C (horizontal line). Although stable in water and aqueous buffer solutions, methylene chloride extraction of plasma treated with compound 2 showed complete conversion of 2 into taxol (1) within 10 minutes (curve, 20% of total recovered). Figure 2 A illustrates a transmission electron micrograph (TEM) of self-assembled helical fibrous nanostructures of taxol-2'-MPT, i.e., compound 2 in buffered solutions (2,1 mM in 100 mM PBS) above the critical aggregation concentration (CAC) of this compound using a negative phosphotungstate stain and a magnification of x25000. The inset shows a portion of one of the fibrils further magnified to illustrate the helical nature of the structure. Figure 2 B illustrates a transmission electron micrograph (TEM) of self-assembled spherical nanostructures of taxol-2'-MPT (compound 2) in unbuffered solutions (2,1 mM in H20) above the critical aggregation concentration (CAC) of this compound using a negative uranyl acetate stain and a magnification of x45000. <Desc/Clms Page number 7> Figure 3 illustrates a tubulin polymerizationdepolymerization measurements with negative control (triangles), positive taxol control (diamonds), and taxol-2'-MPT, i.e., compound 2 (dots). Calcium chloride promoted depolymerization is suppressed by taxol but not by taxol-2'-MPT. Figure 4 illustrates the relative cytotoxicities of taxol-2'-MPT (compound 2) and taxol against a variety of cell lines. Figure 5 illustrates the efficacy of taxol-2'MPT in lung tumor xenograft nude mouse model: 5% dextrose (triangles), taxol (diamonds), and taxol-2'MPT (dots). Detailed Description: The synthesis, physical properties, and pharmaceutical profiles of water soluble onium salts EMI7.1 of taxo-diterpenoid-C",2-O-aza-arenes are described. Synthesis of Taxol-2'-MPT: Taxol-2'-MPT (methylpyridinium tosylate), compound 2, was synthesized according to the method of T.Mukaiyama, Angewandte Chemie 1979, 18(18), 707- 808, incorporated herein by reference. Taxol (10 mg, 0.012 mM), from NaPro Biochemicals, Boulder CO, USA, was dried by azeotropic distillation with toluene (2x1.0 mL) and then dissolved in methylene chloride (0.4 mL) and treated sequentially under an atmosphere of dry argon, with freshly distilled triethylamine (5 microL, 0.04 mM, 3 equivalents) and 2-fluoro-1- methylpyridinium tosylate (5 mg, 0.018 mM, 1.5 equivalents) Aldrich Chemicals, and allowed to stir ,at ambient temperature for 30 minutes. The clear colorless solution rapidly turned to a clear pale yellow. The course of the reaction was monitored <Desc/Clms Page number 8> through thin layer chromatography (TLC) (E. Merck RP-18 silica, 65 tetrahydrofuran : 35 water, UV/phosphomolybdic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC (Rf 0.8). Purification via reverse phase high pressure liquid chromatography (HPLC) (C18 column, 1mM NH40Ac pH 6.5 buffer/ methanol gradient, 1.5 mL/ min. UV) to give, after removal of solvent in vaccuo, pure taxol-2'-MPT (2) (12 mg, 93% yield) as a white amorphous solid. All spectroscopic data (H NMR and HRMS) were in accord with the structure assigned to 2. H NMR (CDC13, 125 MHz) #: 1.055 (s, 3 H, C17-H), 1.083 (s, 3 H, C19-H), 1.724 (s, 3 H, C19-H), 1.858 (m, 1 H, C6-6H), 1.913 (s, 3 H, CH3-Ph), 2.193 (s, 3 H, C10-OC(O)CH3), 2.514 (m, 1 H, C6-aH), 3.663 (d, 1 H, J = 7.0 Hz, C3-H), 4.110 (d, 1 H, J = 8.5, C20-BH, A of AB), 4.133 (s, 3 H, N-CH3), 4.230 (d, 1 H, J = 8.5, C20-aH, B of AB), 4.315 (dd, 1 H, J = 8.7,10.7, C7-H), 4.901 (dd, 1 H, J = 1.0, 7.7, C5-H), 5.501 (d, 1 H, C2-H, J = 7.0), 5.702 (bt, 1 H, C2'-H, J = 8.0), 5.951 (dd, 1 H, C13-H, J = 1.0, 8.0), 6.120 (bd, 1 H, C3'-H, J = 10.0), 6.181 (s, 1 H, C10-H) 7.702 (t, 1 H, N-H, J = 7.5), 7.33 - 7.45 (m, 8 H, Ar-H), 7.56 - 7.62 (m, 4 H, Ar-H), 7.56 - 7.62 (m, 4 H, Ar-H), 7.68 - 7.75 (m, 4 H, Ar-H), 8.00 - 8.05 (m, 1 H Ar-H), 8.23 - 8.28 (m, 1 H, Pyr-H), 8.41 (m, 1 H, Pyr-H). IR (neat, KC1 plate) cm-1: 3640 - 3120 (bm), 3030-2870 (bm), 2320 (m), 1720 (s), 1630 (m), 1560 (m), 1500 (m), 1360 (s), 1160 (m), 1070 (m), 700 (m). UV/Vis (CHC13) nm: 254,280. FAB HRMS: calc for C53H57O14N2: 945.3810; found: 945.3810 The molecular structures of taxol, compound 1, and of taxol-2'-MPT, compound 2, are illustated in Scheme 1A. The synthesis of taxol-2'-MPT is illustrated in Scheme 1B. <Desc/Clms Page number 9> EMI9.1 Schemes 1A and 1B Synthesis of Taxol-7-MPT: The synthesis of taxol-7-MPT differed only slightly from the synthesis of Taxol-2'-MPT. Taxol (10 mg, 0.012 mM), from NaPro Biochemicals, Boulder CO, USA, was dissolved in methylene chloride (2.0 mL) and treated sequentially with triethylamine (67 microL, 0.48 mM, 40 equivalents) and 2-fluoro-1- methylpyridinium tosylate (34 mg, 0.12 mM, 10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. Purification via reverse phase high pressure liquid chromatography (HPLC) gave pure taxol-2'-MPT (2) (12 mg, 93% yield) as a white amorphous solid. The Rf of taxol-7-MPT is about 0.3 minutes less than the Rf of taxol-2'-MPT. The yield was 11 mg or 85%. Spectroscopic data (H NMR and HRMS) were as expected. Synthesis of Taxol-bis-2',7-MPT: The synthesis of taxol-bis-2',7-MPT differed from the synthesis of Taxol-7-MPT only with respect <Desc/Clms Page number 10> to reaction time. Taxol (10 mg, 0.012 mM), from NaPro Biochemicals, Boulder CO, USA, was dissolved in methylene chloride (2.0 mL) and treated sequentially with triethylamine (67 microL, 0.48 mM, 40 equivalents) and 2-fluoro-1- methylpyridinium tosylate (34 mg, 0.12 mM, 10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 18 hours. Purification via reverse phase high pressure liquid chromatography (HPLC) gave pure taxol-2'-MPT (2) (12 mg, 93% yield) as a white amorphous solid. The Rf of taxol-bis-2',7-MPT is about 0.3 minutes less than the Rf of taxol-2'-MPT. The yield was 13 mg or 85%. Spectroscopic data (H NMR and HRMS) were as expected. Alternative synthetic schemes based upon the method of T. Mukaiyama (Anqewandte Chemie 1979, 18(18), 707-808) using a variety of onium salts of 2halogenated aza-arenes for derivatizing either the 2' or the 7 positions of taxol are illustrated in the following scheme: EMI10.1 <Desc/Clms Page number 11> The synthesis of taxol-bis-2',7-MPT differed from the synthesis of Taxol-7-MPT only with respect to reaction time. Taxol (10 mg, 0.012 mM), from NaPro Biochemicals, Boulder CO, USA, was dissolved in methylene chloride (2.0 mL) and treated sequentially with triethylamine (67 microL, 0.48 mM, 40 equivalents) and 2-fluoro-l- methylpyridinium tosylate (34 mg, 0.12 mM, 10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 18 hours. Purification via reverse phase high pressure liquid chromatography (HPLC) gave pure taxol-2'-MPT (2) (12 mg, 93% yield) as a white amorphous solid. The Rf of taxol-bis-2',7-MPT is about 0.3 minutes less than the Rf of taxol-2'-MPT. The yield was 13 mg or 85%. Spectroscopic data (H NMR and HRMS) were as expected. Alternative synthetic schemes based upon the method of T. Mukaiyama (Angewandte Chemie 1979, 18(18), 707-808) using a variety of onium salts of 2halogenated aza-arenes for derivatizing either the 2' or the 7 positions of taxol are illustrated in the following scheme: <Desc/Clms Page number 12> Stability measurements and Kinetics of Taxol release: Due to a difference in retention time using our standard HPLC conditions (see Fig. 1) and differing ultraviolet absorption maxima (l280/l254 = 1. 6 for 2 and 0.3 for 1), the stability of 2 was easily assayed by HPLC (Fig. 1). In all of the ensuing studies, the only degradation products detected were taxol and the pyridinone that results from hydrolysis of pyridinium salts (Fig. lc.). Taxol-2'-MPT appears completely stable in the solid state in a temperature range of -80 t to 25 C regardless of the presence of an inert atmosphere. In water, 5% dextrose, and 1.5% saline 2 is stable for several days but begins to exhibit slow degradation after 4 days. In phosphate buffered saline (PBS) or ammonium acetate - phosphate buffer systems of pH 6.0 to 7.3,2 is stable at 25 C for over 21 days. Taxol-2'-MPT (2) is, however, unstable in 5% HC1 (pH 1.1) and brine. Most significantly, 2 breaks down rapidly when incubated at 37 C with human plasma. This result suggests the presence of factors within plasma that initiate the degradation of 2 to taxol. Since taxol has been shown to bind to albumin to the extent of ca. 85% in sera, it is suspected that basic lysine residues on this protein may initiate breakdown. The kinetics of taxol release from taxol-2'-MPT (2) in various aqueous solutions at 25 C is shown in Figure 1. In sterile water, pH 6.2 phosphate buffered saline, or 5% dextrose, no taxol release is observed over a period of 11 minutes, as shown by the horizontal line. Although stable in water and aqueous buffer solutions, methylene chloride extraction of plasma treated with compound 2 showed <Desc/Clms Page number 13> complete conversion of 2 into taxol (1) within 10 minutes, as shown by the curved line. Under these conditions 20% of total is recovered. More particularly, Taxol-2'-MPT (2) was dissolved in the aqueous system with the aid of sonication for five minutes. Aliquots were then removed at the times shown and partitioned into methylene chloride to quench the reaction. Samples were then analyzed using a Waters Maxima HPLC instrument equipped with an autoinjector (3.9x300 mm C18 column equipped with a precolumn. The flow rate was 1.5 mL/minute. The eluent gradient A-B extended over 30 minutes. "A" was 80% 80mM ammonium acetate, pH 6.0. "B" was 100% methanol. An ultraviolet diode array detector was employed. The ratio of compound 2 (Rf 16.2 min. ) to taxol (compound 1, Rf 16.8 min. ) remaining was determined from the relative areas of the peaks after normalization with previously determined calibration curves. <Desc/Clms Page number 14> Solubility Measurements: The solubility and partition coefficient data for 2 and taxol were determined using an HPLC method. EMI14.1 <tb> <tb> Taxol <SEP> 2'-MPT-taxol <tb> EMI14.2 Solubility in < 1.5 x 10- M 1.7 x 10 EMI14.3 <tb> <tb> Water <tb> Partition <SEP> > <SEP> 10000 <SEP> 50 <SEP> <tb> Coefficient <tb> [octanol] <SEP> /[water] <tb> Solubility was found by forming a solution in water with the aid of sonication for five minutes, centrifugation of the samples, and injection of the supernatant. The values reported were normalized using calibration curves constructed for both compounds by preparing known concentrations in the range 1 x 10-6 to 1 x 10-3 M in methylene chloride and subjecting to HPLC analysis under the same conditions. One should note that the solubility of taxol is at the detection limit of the instrument and thus represents an upper limit. The solubility of 2 was found at a concentration at which the solution was clear (see below) and thus represents a lower limit. Compound 2 exhibited similar solubilities for various buffer systems in the pH range 6.2 to 7.4. Partition coefficients were determined by dissolving the compound in the organic phase, shaking the resulting solution with water for ten minutes, and analyzing each phase by HPLC as above. No degradation of 2 was noted during these studies. <Desc/Clms Page number 15> These data clearly show that 2 is significantly more soluble in water than the parent taxol. The solubility demonstrated in a range of aqueous systems is higher than the clinically relevant dosages (3 to 30 mM). Self-Assembling Structures: While the water solutions of 2 were optically clear at all concentrations examined, buffered solutions of concentrations greater than 1 x 10-3 M exhibited a haze to the naked eye and diffuse scattering of monochromatic light. Ultraviolet spectroscopic absorption measurements at 340 nm (Fig. 3) showed an exponential increase in optical density (OD) above a critical concentration of 4 x 10-4 M, a result characteristic of macromolecular structure in solution. Transmission electron microscopy (TEM) confirmed the presence of supramolecular structures in these solutions. Uniform aggregates of fibrillar structure (Fig. 2a) with helical conformations were observed. These structures exhibited varying (up to 800 A) lengths but consistent diameter of ca. 80 with a helical twist of about 7. Additionally, freshly sonicated solutions of 2 showed the presence of spherical structures (Fig. 2b) with diameters of about 50 A. It is likely that the long term stability of these solutions is due, at least in part, to stabilization provided by this structured environment. Microtubule polymerization-depolymerization measurements: Microtubule polymerization-depolymerization measurements (Fig. 3) with taxol-2'-MPT (2) were very similar to GTP-saline controls and drastically different from taxol. Compound 2 does not appear to <Desc/Clms Page number 16> bind to tubulin in the manner of taxol. In the buffered aqueous environment of this assay, 2 is not converted to taxol and thus does not affect the tubulin-microtubule equilibria. Taxol, recovered from human plasma treated with 2, exhibited the expected microtubule stabilization, indicating that 2 does act as a prodrug for taxol. Tubulin polymerization-depolymerization measurements are illustrate in Figure 3. Negative controls are shown with triangles; positive taxol controls are shown with diamonds; and taxol-2'-MPT, i.e., compound 2 is shown with dots. The measurements indicate that calcium chloride promoted depolymerization is suppressed by taxol but not by taxol-2'-MPT. More particularly, measurement were performed in 96 well plates at 37 C following the protocol of R. Merlock and W. Wrasidlo (Analytical Biochemistry 1993, in press). Calcium chloride addition is indicated by the arrow. In each case, 1.0 mM GTP was used to promote the initial polymerization of tubulin. Negative control employed tubulin (1.0 mg/mL) alone, CaCl2 (0,25 mM) added after 20 minutes. Positive taxol control employed tubulin (1.0 mg/mL) with taxol (10-6M) and CaCl2 (0.25mM) added after 20 minutes. The experimental taxol-2'-MPT employed tubulin (1.0 mg/mL) with taxol-2'-MPT (10-6) and CaClz (0.25 mM) added after 20 minutes. Turbidity was measured as optical density at 340 nm using a microplate reader (Molecular Devices Thermomax). Toxicity Measurements: Compound 2 was tested for its cytotoxicity against a cell line panel including leukemia, ovarian, lung, and breast carcinoma cells (Fig. 4). The differential cytotoxicity profiles for 2 and <Desc/Clms Page number 17> taxol were similar, although some differences were noted. Both compounds exhibited IC50 values ranging from 10-5 to 10-12 M with means close to one nanomolar. Normal cells had cytotoxicity levels three to four orders of magnitude below mean values. Extremely high cytotoxicity levels were recorded for human leukemia, metastatic melanoma and cervical carcinoma. As expected for a prodrug of taxol in the cellular environment, 2 shows the same remarkable tumor cell selectivity and cell line specificity as taxol. The relative cytotoxicities of taxol-2'-MPT (compound 2) and taxol against a variety of cell lines are illustrated in Figure 4. More particularly, cells were plated on 96 well plates with the following controls: no cells and toxic control (1 x 10-3 M SDS). The drug was added to the first set of wells and diluted via standard dilution method from the stock. Plates were incubated at 37 C, 5% CO2 in sterile air in an humidified incubator for 72 hours. An aliquot of 50 L of a solution of 2,3-bis(methoxy-4-nitro-5-sulfophenyl)-5- [(phenylamino)carbonyl]-lH-tetrazolium hydroxide (XTT), 1 mg mL-1, in phosphate buffered saline (PBS, 100mM) was added to the wells. In the presence of viable cells, this colorless clear media is enzymatically altered to give a pink coloration. The plates were read at 450 nm using a plate reader. Percentage cytotoxicity was calculated using the formula: %C = 1 - (OD toxin)(OD growth control)-1(100). Efficacy of taxol-2'-MPT in lung tumor xenoqraft: The encouraging in vitro data obtained with taxol-2'-MPT (2) prompted us to study its in vivo action using nude mice inflicted with human lung carcinoma xenografts (Fig. 1). The samples of 2 used <Desc/Clms Page number 18> for this study were formulated in sterile PBS without Cremaphor, indicating the suitability of this compound for simple bolus administration. Preliminary data shows that the control of tumor growth exhibited by 2 is at least comparable to that of taxol and significantly (0.001 p-value, multiple linear regression model) different from controls. These results provide a reasonable indication that 2 is converted rapidly to taxol in vivo and should thus exhibit pharmacology similar to taxol. Indeed, in metabolic study using tritiated 2, only 5% of the compound was excreted through the kidneys, a result that is completely in accord with the behavior of taxol. The efficacy of taxol-2'-MPT in lung tumor xenograft nude mouse model is illustrated in Figure 5. The tumor model was generated from an ATCC A549 non-small cell lung adenocarcinoma cell line that was maintained under the standard cell proliferation conditions (37 C, 5% carbon dioxide in sterile air). Hemocytometer counted cells suspended in Hanks medium (Gibco, Grand Island NY) were implanted S.C. (106 cells in 0.4 mL per tumor volume determined using the equation (length)(width ) 2/2. The test compounds (1.0 microM) were injected I.P. on day 1,3, and 7 using the following media: control. 5% dextrose in water (D5W), triangles; taxol, suspended in Cremaphor/D5W (5/95,18.0 mg/kg of animal weight), diamonds; and taxol-2'-MPT, dissolved in D5W (23.9 mg/kg of animal wight), dots. The procedures used for the maintenance of tumors and the experimental details were according to protocols set forth by the Developmental Therapeutics Program, National Cancer Institute, viz. , National Cancer Institute Cancer Chemotherapy Reports, 3 (1972). <Desc/Clms Page number 19> Mechanisms of taxol release: The mechanism of acid catalyzed taxol (1) release from taxol-2'-MPT is illustrated in Scheme 2. EMI19.1 Scheme 2 However, the release of taxol from taxol-2'-MPT can also be catalyzed by serum protein and by proteins having nucleophilic groups. When contacted with serum protein, taxol-2'-MPT is observed to displace its MPT group and form a protein:taxol intermediate. Dialysis of the protein:taxol intermediate indicates a dissociation period of hours or days. Displacement of the MPT group by serum proteins seems to be specific for such serum proteins. Tested non-serum proteins seemed to lack this activity. In particular, immunoglobulins and serum albumen seem to be particularly effective displacing the MPT group and forming protein:taxol intermediates. The precise nature of the bonding between the protein and taxol has not been characterized. Scheme 2 illustrates alternative pathways for MPT release. Discussion Taxol-2'-MPT has proven to be a remarkably stable compound in most aqueous media. It is probable that this stability is conferred upon 2 by the facile formation of supramolecular aggregates, a process that is probably driven by the amphiphillic <Desc/Clms Page number 20> nature of the compound. The stability, water solubility, and lack of cytotoxicity of taxol-2'-MPT makes this class of compound an ideal a prodrug for taxol and memetics of taxol. While essentially completely stable in aqueous media at physiological pH and ion strength, the compound rapidly discharges taxol in sera. This profile is ideal for a clinically useful prodrug to taxol. It is possible that these properties should allow the formulation of taxol-2'-MPT (2) without the use of Cremaphor or ethanol. <Desc/Clms Page number 21> EMI21.1 <Desc/Clms Page number 22> Synthetic Methods Preparation of 7-TES deacetylbaccatin III (4) EMI22.1 7-TES deacetylbaccatin III (4). To a solution of 10deacetylbaccatin III (3, 3.0 g, 5 .51 mmol, Indena Corpation, Italy) in pyridine (250 mL) was added chlorotriethylsilane ( 18.5 mL, 110 mmol) dropwise. The resulting solution was stirred at 25 C for 17 hours. After dilution with diethylether (750 mL), the solution was washed with aqueous CUS04 (3 x 200 mL) and brine (200 mL). The organic layer was dried (MgSO4), concentrated, and purified by flash chromatography (silica, 35 -50% ethylacetate in petroleum ether) to give alcohol 4 (3.39 g, 91%) as a white solid. Physical Data for 7-TES deacetylbaccatin III (4). Rf= 0.32 (silica, 50% ethylacetate in hexanes); IR (thin film) #max 3464, 2954, 2282, 1710, 1453, 1362, 1271, 1242, 1105, EMI22.2 994 cl-1; lH NMR (500 MHz, COC13) 5 8.06 (dd, J= 8.0, 0.9 Hz, 2 H, Bz), 7.57 (t, J= 7.9 Hz, 1 H, Bz), 7.44 (t, J= 7.9 Hz, 2 H, Bz), 5.56(d,J=7.OHz,lH,2-H),5.14(d,J=l.9Hz,1H,10-H),4.92 (d, J = 9.5 Hz, 1 H, 5-H), 4.84-4.78 (m, 1 H, 13-H), 4.37 (dd, J= <Desc/Clms Page number 23> 10.6, 7.0 Hz, 1 H, 7-H), 4.27 (d, J= 8.5 Hz, 1 H, 20-H), 4.25 (d, J = 1.9 Hz, 1 H, 10-OH), 4.12 (d, J= 8.5 Hz, 1 H, 20-H), 3.91 (d, J= 7.0 Hz, 1 H, 3-H), 2.48-2.40 (m, 1 H, 6-H), 2.25 (s, 3 H, Me), 2.25-2.17 (m, 2 H, 14-CH2), 2.04 (s, 3 H, Me), 1.90-1.82 (m, 1 H, 6-H), 1.70 (s, 3 H, Me), 1.03 (s, 6 H, Me, Me), 0.90 (t, J= 8 Hz, 9 H, Si(CH2CH3)3), 0.58-0.42 (band, 6 H, Si(CH2CH3)3); 13CNMR EMI23.1 (125 MHz, CDCl3) 8 210.3, 170.8, 167.0, 141.8, 135.1, 133.6, 130.1, 129.4, 128.6, 84.2, 80.7, 78.8, 76.5, 74.8, 74.6,72.9, 67.9, EMI23.2 z7.9, 47.0, 42.7, 38.6, 37.2, 26.8, 22.6, 19.5, 15.2, 9.9, 6.7, 5.1; FAB HRMS (NBA/ CsI) m / e 791.2251, M + Cs+ calcd for C35H50010Si 791.2228. Preparation of enone 5 EMI23.3 Enone 5. To a solution of 7-TES deacetylbaccatin III (4, 1.5 g, 2.28 mmol) and 4-methylmorpholine N-oxide (NMO, 240 mg, 2.05 mmol) in CH2C12 (5 mL) was added 4 molecular sieves (200 mg) and the suspension was stirred at 25 C for 10 minutesutes. A catalytic amount of tetrapropylammonium perruthenate from Aldrich Chemical Company Inc. (TPAP, 40 mg, 0.11 mmol) was added by portions and the reaction <Desc/Clms Page number 24> mixture was stirred at 25 C for 0.5 hours. Small amounts of 4methylmorpholine N-oxide and TPAP were added alternatively at 0.5 hour intervals until the starting material was consumed to the extent of ca. 95% by TLC. The reaction mixture was filtered through silica gel, eluted with CH2C12 (100 mL), and concentrated to give enone 5 (1.44 g, 96%) as a white solid. Physical Data for Enone 5. Rf = 0.5 (silica, 50% ethylacetate in hexanes); IR (thin film) vmax 3446, 2957, 2882, 1726,1672, 1456,1367, 1243, 1106 cm-1; 1H NMR (500 MHz, CDC13) S 8.05 (dd, J= 8.0, 1.0 Hz, 2 H, Bz), 7.61 (t, J= 7.5 Hz, 1 H, Bz), 7.45 (t, J = 7.5 Hz, 2 H, Bz), 5.63 (d, J= 7.5 Hz, 1 H, 2-H), 5.30 (d, J= 2.0 Hz, 1 H, 10-H), 4.90 (d, J= 8.0 Hz, 1 H, 5-H), 4.36 (dd, J = 10.5, 7.0 Hz, 1 H, 7-H), 4.31 (d, J= 8.5 Hz, 1 H, 20-H), 4.30 (d, J = 2.0 Hz, 1 H, 10-OH), 4.11 (d, J = 8.5 Hz, 1 H, 20-H), 3.93 (d, J= 7.5 Hz, 1 H, 3-H), 2.92 (d, J= 19.5 Hz, 1 H, 14-H), 2.62 (d,J= 19.5 Hz, 1 H, 14-H), 2.50-2.42 (m, 1 H, 6-H), 2.17 (s, 3 H, Me), 2.08 (s, 3 H, Me), 1.90-1.82 (m, 1 H, 6-H), 1.77 (s, 1 H, 1-OH), 1.70 (s, 3 H, Me),. 1.21 (s, 3 H, Me), 1.14 (s, 3 H, Me), 0.90 (t, J= 8.0 Hz, 9 H, Si(CH2CH 3) 3), 0.60-0.42 (band, 6 H, Si(CH2CH3)3); 13C NMR (125 MHz, CDC13) # 208.2, 198.1, 170.2, EMI24.1 166.8, 156.6, 139.1, 134.0, 130.0, 128.8, 128.8, 84.0, 80.4, 78., 76.2, 75.7, 72.9, 72.8, 58.8, 45.9, 43.4, 42.5, 37.2, 33.0, 21.7, 17.5,13.6, 9.6,6.7, 5.1: FAB HRMS (NBA/ NaI) m / e 657.3070, M + Na+ calcd for C35H48O10Si 657.3095. <Desc/Clms Page number 25> Preparation of triol 6 EMI25.1 Triol 6. To a solution of enone 5 (1.44 g, 2.19 mmol) in MeOH (300 mL) at 0 C was slowly added an aqueous solution ofK2C03 (3.0 g in 32 mL of H?O) . The solution was stirred at 0 C for 2.5 hours. The reaction was then quenched with aqueous NH4C1 ( 150 mL) and the resulting mixture was extracted with CH2C12 (2 x 200 mL). The organic layer was dried (Na2S04), concentrated, and purified by flash chromatography (silica, 35 # 50% ethylacetate in petroleum ether) to give enone 5 (270 mg, 19%) and triol 6 (912 mg, 93% based on 81% conversion). Physical Data for Triol 6. Rf= 0.24 (silica, 50% ethylacetate in hexanes); IR (thin film) #max 3414, 2957, 2881, 1727, 1664, 1370 cm-1; 1H NMR (500 MHz, CDCI3) # 5.23 (d, J= 9.5 Hz, 1 H, 10-H), 4.89 (d, J= 9.5 Hz, 1 H, 5-H), 4.63 (d, J = 9.5 Hz, 1 H, 20-H), 4.56 (d, J= 9.5 Hz, 1 H, 20-H), 4.32 (dd, J= 11.0, 7.0 Hz, 1 H, 7-H), 4.28 (d, J = 2.5 Hz, 1 H, 10-OH), 3.89 (dd, J= 6.5, 4.0 Hz, 1 H,2-H), 3.57 (d, J= 6.5 Hz, 1 H, 3-H), 2.78 (d, J= 19.5 Hz, 1 H, 14-H), 2.58 (d, 4.0 Hz, 1 H, 2-OH), 2.52 (d, J= 19.5 Hz, 1 H, 14-H), 2.49-2.42 (m, 1 H, 6-H), 2.03 (s, 3 H, Me), 1.92- 1.84 (m, 1 H, 6-H), 1.68 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.04 (s, 3 <Desc/Clms Page number 26> H, Me), 0.90 (t, J= 8.0 Hz, 9 H, Si(CH2CH3)3), 0.60-0.40 (band, 6 EMI26.1 H, Si(CH?CH3)3); 13C NMR ( 125 MHz, CDC13) 8 208.9, 198.5, 170.1, 156.7, 138.8, 83.8, 81.2, 77.6, 75.7, 72.8, 72.5, 58.8, 45.8, 43.1, 42.8, 37.3, 32.7, 21.6, 17.5, 13.6,9.7, 6.7, 5.1; FAB HRMS (NBA NaI) m / e 575.2648, M + Na+ calcd for C2sH4409Si 575.2652. Preparation of Carbonate 7 EMI26.2 Carbonate 7. A solution of triol 6 (60.0 mg, 0.109 mmol) in THF (2 mL) was treated with carbonyldiimidazole ( 110.0 mg, 0.678 mmol) and stirred at 40 C for 0.5 hour The reaction mixture was concentrated and redisolved in THF (5 mL). TLC analysis confirmed total consumption of starting material. IN aqueous HCl (5 mL) was added and the resulting solution was allowed to stir for 15 minutes at 25 C. diethylether (25 mL) was added, the organic layer was separated, washed with aqueous NaHC03 (10 mL) and brine (10 mL), dried (MgS04), and concentrated to give carbonate 7 (58 mg, 93%) as a white foam. Physical Data for Carbonate 7. Rf = 0.50 (silica, 35% EMI26.3 ethylacetate in hexanes); IR (thin film) va; 3438, 2957, 2882, <Desc/Clms Page number 27> 1820, 1731, 1685, 1370, 1236 cm-1; 1H NMR (500 MHz, CDC13) 8 5.27 (d, J= 2.5 Hz, 1 H, 10-H), 4.89 (d, J= 9.0 Hz, 1 H, 5-H), 4.60 (d, J = 9.0 Hz, 1 H, 20-H), 4.45 (d, J= 9.0 Hz, 1 H, 20-H), 4.43 (d, J= 6.0 Hz, 1 H, 2-H), 4.33 (dd, J= 10.0, 7.5 Hz, 1 H, 7-H), 4.28 (d, J= 2.5 Hz, 1 H, 10-OH), 3.54 (d, J= 6.0 Hz, 1 H, 3-H), 2.88 (d, J= 20.0 Hz, 1 H, 14-H), 2.75 (d, J = 20.0 Hz, 1 H, 14-H), 2.54- 2.47 (m, 1 H, 6-H), 2.08 (s, 3 H, Me), 2.06 (s, 3 H, Me), 1.92-1.84 (m, 1 H, 6-H), 1.77 (s, 3 H, Me), 1.31 (s, 3 H, Me), 1.15 (s, 3 H, Me), 0.88 (t, J= 8.5 Hz, 9 H, Si(CH2CH3)3), 0.55-0.45 (band, 6 H, EMI27.1 Si(CH¯2CH3)3); 13C NMR (125 MHz, CDC13) 5 208.4, 195.5, 170.5, 154.0, 152.0, 141.2, 88.4, 83.9, 79.8, 79.0, 76.7, 75.7, 71.9, 60.3, 43.0,41.6, 39.8, 37.7, 31.6, 21.5, 17.8, 14.4, 9.7, 6.6, 5.0: FAB HRMS (NBA) m/ e 579.2652, M + H+ calcd for C29H42O10Si 579.2626. Preparation of nButyl-C-2 ester derivative (Alcohol 8) EMI27.2 Alcohol 8. A solution of carbonate 7 (10 mg, 0.0173 mmol) in tetrahydrofuran (1 mL) at -78 C was treated with n-Butyllithium from Aldrich Chemical Company, Inc. (0.087 mL of a 1.6 M solution in hexanes, 0.139 mmol) and <Desc/Clms Page number 28> stirred for 1.0 hour The reaction mixture,was poured into a mixture of diethylether (10 mL) and aqueous NH4CI (5 mL) . The organic layer was separated and the aqueous layer was extracted with diethylether (2 x 5 mL). The combined organic layer was washed with a saturated solution of brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 35 - 50% ethylacetate in hexanes) to give 8 (7.9 mg, 72%) as an amorphous solid. Physical Data for Alcohol 8 . Rf = 0.36 (silica, 35% ethylacetate in petroleum ether) ; IR (film) vmax 3437,2962, 2865,1726, 1671,1367, 1239, 1105 cm-1; 1H NMR (500 MHz, CDC13) # 5.36 (d, J= 6.5 Hz, 1H, 2-H), 5.26 (d, J= 2.5 Hz, 1H, 10- H), 4.89 (br d, J= 8.0 Hz, 1H, 5-H), 4.47 (d, J = 8.0 Hz, 1H, 20-H), 4.32 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.26 (d, J = 2.5 Hz, 1H, 10- OH), 4.15 (d, J= 8.0 Hz, 1H, 20-H), 3.81 (d, J= 6.5 Hz, 1H, 3-H), 2.73 (d, J= 20.0 Hz, 1H, 14-H), 2.57 (d, J= 20.0 Hz, 1H, 14-H), 2.49-2.41 (m, 1H, 6-H), 2.38-2.23 (m, 2H, OCCH2(CH2)2CH3), 2.06 (s, 3H, Me), 2.04 (s, 3H, Me), 1.90-1.82 (m, 1H, 6-H), 1.67 (s, 1H, OH), 1.64 (s, 3H, Me), 1.68-1.52 (m, 2H, OCCH2CH2CH2CH3), 1.41- 1.30 (m, 2H, OC(CH2) 2CH2CH3), 1.19 (s, 3H, Me), 1.07 (s, 3 H, Me), 0.94-0.86 (band, 12H, CH3 of Bu, OSi(CH2CH3)3), 0.58-0.45 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA) m / e 637.3421, M + H+ calcd for C33H52010Si 637.3408. <Desc/Clms Page number 29> Preparation of vinyl-C-2 ester derivative (Alcohols 9 and 10) EMI29.1 Alcohols 9 and 10. -A solution of carbonate 7 ( 111.3 mg, 0.192 mmol) in tetrahydrofuran ( 2 mL) at -78 C was treated with vinyllithium (3.7 mL of a 0.52 M solution in diethylether, 1.92 mmol, prepared from tetravinyltin and EMI29.2 nBuyllithium: methodology from th'akefield, B.J. Organolithium Methods, Academic Press: London, 1988, p. 46) and stirred for 2.25 hour The reaction mixture was poured into a mixture of CH2Cl2 (20 mL) and aqueous NH4Cl (10 mL), the organic layer was separated, and the aqueous layer was extracted with CH2Cl2 (3 x 10 mL). The combined organic layer was washed with brine (15 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 30 # 50% ethylacetate in <Desc/Clms Page number 30> petroleum ether) to give 9 (60.0 mg, 52%); and 10 (25.7 mg, 24% ) as white foams. Physical Data for Alcohol 9. Rf = 0.52 (silica, 50% ethylacetate in hexanes); IR (film) vmax 3442,2956, 2882, 1727, 1672, 1407, 1368, 1243, 1182, 1110, 1050, 986, 826, EMI30.1 7 3 6 cm-1; 1 H NMR ( 5 00 MHz, CDC13) 8 6 . 51 (dd, J= 17 .0, 1.0 Hz, 1H, vinyl H), 6.13 (dd, J= 17.0, 10.5 Hz, 1H, vinyl H), 6.00 (dd, J = 10.5, 1.0 Hz, 1H, vinyl H), 5.45 (br d, J= 6.5 Hz, 1H, 2-H), 5.30 EMI30.2 (d, J = 2.5 Hz, 1 H, 10-H), 4 .91 (br d, J = 9.5 Hz, 1 H, 5 -H ) , 4.44 (d, J= 8.5 Hz, 1H, 20-H), 4.35 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.30 (d, J= 2.5 Hz, 1H, 10-OH), 4.14 (d, J = 8.5 Hz, 1H, 20-H), 3.88 (d, J= 6.5 Hz, 1H, 3-H), 2.79 (d, J = 20.0 Hz, 1H, 14-H), 2.61 (d, J= 20.0 Hz, 1H, 14-H), 2.48 (ddd, J = 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.09 (s, 3H, Me), 2.08 (s, 3H, Me), 1.89 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.72 (s, 1H, OH), 1.68 (s, 3H, Me), 1.22 (s, 3H, Me), 1.12 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.62-0.46 EMI30.3 (band, 6H, OSi(CH C H 3 ) 3 ); FAB HRMS ( NBA / CSI) m / e 739.1925, M + Cs+ calcd for C31H46O10Si 739.1915. Physical Data for Alcohol 10. Rf = 0.24 (silica, 50% ethylacetate in hexanes); IR (film) #max 3439, 2955, 2881, 1711, 1671, 1409, 1365, 1188, 1115, 980, 833, 735 cm-1; 1H NMR (500 MHz, CDC13) 8 6.48 (br d, J = 17.0 Hz, 1H, vinyl H), 6.10 (dd, J= 17.0, 10.5 Hz, 1H, vinyl H), 5.97 (br d, J= 10.5 Hz, EMI30.4 1H, vinyl H), 5.47 (br d, J= 6.0 Hz, 1H, 2-H), 5.25 (d, J= 2. Hz, 1H, 10-H), 4.75 (dd, J = 9.5, 3.5 Hz, 1H, 5-H), 4.38 (d, J = 8.5 Hz, 1H, 20-H), 4.30 (d, J= 2.5 Hz, 1H, 10-OH), 4.24 (d, J= 8.5 Hz, 1H, 20-H), 3.90 (dd, J= 11.5, 6.0 Hz, 1H, 7-H), 3.28 (d, J= 19.5 Hz, <Desc/Clms Page number 31> 1H, 14-H), 3.24 (d, J= 6.0 Hz, 1H, 3-H), 3.06 (br s, 1H, OH), 2.58 (d, J= 19.5 Hz, 1H, 14-H), 2.38 (ddd, J= 14.5,9.5,6.0 Hz, 1H, 6H), 2.07 (s, 3H, Me), 1.98 (ddd, J= 14.5, 11.5, 3.5 Hz, 1H, 6-H) 1.87 (s, 1H, OH), 1.61 (s, 3H, Me), 1.23 (s, 3H, Me)', 1.13 (s, 3H, Me), 0.90 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.59-0.45 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m / e 697.1802, M + Cs+ calcd for C29H4409Si 697.1809. Preparation of 2-Furyl-C-2 ester derivative (Alcohol 11) EMI31.1 Alcohol 11. A solution of carbonate 7 (46 mg, 0.0795 mmol) in tetrahydrofuran (3 mL) at -78 C was treated with 2furyllithium (4 mL of a 0.47 M suspension in diethylether, 1.88 mmol, prepared from furan (Aldrich Chemical Company, Inc.) and n-Butyllithium (Aldrich Che'mical Company, Inc. ); methodology from Ramanathan, V.; Levine, R. J. Org. Chem. 1962, 27, 1216) and stirred for 10 minutesutes The reaction mixture was poured into a mixture of CH2C12 (15 mL) and aqueous NH4CI (20 mL). The organic layer was separated and <Desc/Clms Page number 32> the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine (10 mL), dried (MgS04) and concentrated to give 11 which was taken into the next step without further purification. Physical Data for Alcohol 11. Rf = 0.38 (silica, 20% ethylacetate in petroleum ether) ; IR (film) vmax 3442,2956, 2882, 1727, 1672, 1468, 1300,1240, 1110, 1007, 733 cm-1; 1H NMR (500 MHz, CDC13) 8 7.66-7.64 (m, 1H, furan), 7.24 (br d, J= 3.5 Hz, 1H, furan), 6.58 (dd, J= 3.5, 1.5 Hz, 1H, furan), 5.55 (d, J EMI32.1 = 6.5 Hz, 1H, 2-H), S.31 (d, J = 2.0 Hz, 1H, 10-H), 4.92 (br d, j = 9.0 Hz, 1H, 5-H), 4.43 (d, J = 8.5 Hz, 1H, 20-H), 4.37 (dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.32 (d, J= 2.0 Hz, 1H, 10-OH), 4.18 (d, J = 8.5 Hz, 1H, 20-H), 3.93 (d, J = 6.5 Hz, 1H, 3-H), 2.88 (d, J 20.0 Hz, 1H, 14-H), 2.63 (d, J= 20.0 Hz, 1H, 14-H), 2.55-2.37 (m, 1H, 6-H), 2.15 (s, 3H, Me), 2.09 (s, 3H, Me), 1.93-1.87 (m, 1H, 6H), 1.81 (s, 1H, OH), 1.71 (s, 3H, Me), 1.23 (s, 3H, Me), 1.15 (s, 3H, Me), 0.93 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.62-0.42 (band, EMI32.2 6H, OSi(CH?CH3)3); FAB HRMS (NBA / Nal) m / e 669.2717, M + Na+ calcd for C33H46O11Si 669.2707. <Desc/Clms Page number 33> Preparation of 2-thiophenyl-C-2 ester derivative (Alcohol 12) EMI33.1 Alcohol 12. A solution of carbonate 7 (50.0 mg, 0.0864 mmol) in tetrahydrofuran (5 mL) at -78 C was treated with 2-thienyllithium from Aldrich Chemical Company, inc. (1.30 mL of a 1.0 M solution in tetrahydrofuran, 1.30 mmol) and stirred for 0.5 hour The reaction mixture was poured into a mixture of diethylether (10 mL) and aqueous NH4Cl (5 mL) . The organic layer was separated and the aqueous layer was extracted with diethylether (2 x 10 mL). The combined organic layer was washed with brine (10 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10 # 35% ethylacetate in hexanes) to give 7 (16.5 mg, 33%), 12 (36.8 mg, 96% based on 67% conversion) as an amorphous solid. Physical Data for Alcohol 12. Rf = 0.56 (silica, 50% ethylacetate in hexanes); IR (film) #max 3403, 2945, 2881. 1717,1669,1520,1413,1360,1248,1078; 1H NMR (500 MHz, CDC13) 8 7.84 (dd, J= 3.5, 1.0 Hz, 1H, thiophene) , 7.64 (d, J= 1.0, <Desc/Clms Page number 34> 5.0 Hz, 1H, thiophene), 7.14 (dd, J = 5.0, 3.5 Hz, 1H, thiophene), 5.53 (br d, J = 6.5 Hz, 1H, 2-H),5.29 (d, J = 2.5 Hz, 1H, 10-H), 4.90 (br d, J = 7.5 Hz, 1H, 5-H), 4.44 (d, J = 8.5 Hz, 1H, 20-H), 4.35 (dd, J = 10.5 Hz, 6.5 Hz, 1H, 7-H), 4.29 (d, J= 2.5 Hz, 1H, 10-OH), 4.19 (d, J= 8.5 Hz, 1H, 20-H), 3.90 (d, J= 6.5 Hz, 1H, 3H), 2.89 (d, J = 19.5 Hz, 1H, 14-H), 2.62 (d,J= 19.5 Hz, 1H, 14H), 2.49-2.43 (m, 1H, 6-H), 2.15 (s, 3H, Me), 2.07 (s, 3H, Me), 1.92-1.84 (m, 1H, 6-H) , 1.73 (s, 1H, OH), 1.71 (s, 3H, Me), 1.21 (s, 3H, Me), 1.13 (s, 3H, Me), 0.91 (t, J = 8.0 Hz, 9H, OSi(CH?CH3)3), 0.56-0.49 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA) m/ e 663.2655, M + H- calcd for C33H46O10SSi 663.2659. Preparation of 3-thiophenyl-C-2 ester derivatives (Alcohol 13 and 14) EMI34.1 <Desc/Clms Page number 35> EMI35.1 Alcohols 13 and 14. A solution of carbonate 7 (107.9 mg, 0.186 mmol) in tetrahydrofuran (6.2 mL) at -78 C was treated with 3-thienyllithium (2.76 mL of a 0.41 M solution in EMI35.2 diethylether : tetrahyd?ofu9an. :"h±lanes (4.5 : 1 : 2), 1.13 mmol, prepared from 3-bromothiophene and n-Butyllithium; methodology from Camici, L.; Ricci, A. ; Taddei, M. Tetrahedron Lett. 1986, 27, 5155),,and -stirred for 1.5 hour The reaction mixture was poured into a ''Mixture of CH2C12 (15 mL) and aqueous NH4Cl (20 mL), the organic layer was separated, and the aqueous layer was extracted with CH2C12 (2 x 10 mL). The combined organic layer was washed with brine (10 mL) , dried (MgS04), concentrated, and purified by flash chromatography (silica, 20 # 30% ethylacetate in hexanes) to give 7 (16.9 mg, 16%), 13 (87.0 mg, 83% based on 84% conversion), and hydrolyzed C4 acetate 14 (C4-hydrolyzed side product, 9.7 mg, 10% based on 84% conversion) as amorphous solids. Physical Data for Alcohol 13. Rf = 0.74 (silica, 50% ethylacetate in hexanes), 0.41 (silica, 10% ethylacetate in benzene, 3 elutions) ; IR (thin film) #max 3442, 3110, 2956, 2882, 1725, 1672, 1410, 1368, 1244, 1198, 1101, 988, 825, <Desc/Clms Page number 36> 744 cm-I; 1H NMR (500 MHz, CDC13) s 8.18 (dd, J = 3.0, 1.2 Hz, 1H, thiophene), 7.54 (dd, J = 5.0, 1.2 Hz, 1H, thiophene), 7.37 (dd, J= 5.0,3.0 Hz, 1H, thiophene), 5.56 (dd, J= 6.5, 1.0 Hz, 1H, 2-H), 5.31 (d, J= 2.5 Hz, 1H, 10-H), 4.92 (dd, J = 7.5, 2.0 Hz, 1H, 5-H), 4.40-4.34 (m, 2H, 20-H, 7-H), 4.31 (d, J = 2.5 Hz, 1H, 10OH), 4.15 (d, J= 8.5 Hz, 1H, 20-H), 3.93 (d, J = 6.5 Hz, 1H, 3-H), 2.88 (d, J= 20 Hz, 1H, 14-H), 2.63 (dd, J= 20.0, 1.0 Hz, 1H, 14H), 2.47 (ddd, J = 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.18 (s, 3H, Me), 2.10 (s, 3H, Me), 1.89 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.81 (br s, 1H, OH), 1.72 (s, 3H Me), 1.23 (s, 3H, Me), 1.15 (s, 3H, Me), 0.93 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.62-0.48 (band, 6H, EMI36.1 Si(CH2CH3)3); FAB HRMS (NBA / Csl) m / e 795.1640, 1 + Cs- calcd for C33H46O10SSi 795.1635. Physical Data for Alcohol 14: Rf = 0.54 (silica, 50% ethylacetate in hexanes); IR (thin film) vmax 3437, 3108, 2955,2880,1709,1674,1605,1520,1410,1360,1258,1194, 1103, 1004,829, 744 cm-1; 1H NMR (500 MHz, CDC13) # 8.15 (dd, J = 3.0,1.0 Hz, 1H, thiophene), 7.49 (dd, J= 5.0,1.0 Hz, 1H, thiophene), 7.35 (dd, J= 5.0, 3.0 Hz, 1H, thiophene), 5.59 (d, J= EMI36.2 6.0 Hz, 1H, 2-H), 5.fla (d, J= 2.5 Hz, 1H, 10-H), 4.73 (dd, J= 9.5, 3.5 Hz, 1H, 5-H), 4.40 (d, J= 8.5 Hz, 1H, 20-H), 4.32 (d, J= 2.5 Hz, 1H, 10-OH), 4.15 (,d, J= 8.5 Hz, 1H, 20-H), 3.92 (dd, J= 11.5, 6.0 Hz, 1H, 7-H), 3.44 (d, J= 19.5 Hz, ,1H, 14-H), 3.30 (d, J= 6.0 Hz, 1H, 3-H), 2.91 (br s, 1H, OH), 2.61 (d, J= 19.5 Hz, 1H, 14-H) , 2.38 (ddd, J= 14.5, 9.5, 6.0 Hz, 1H, 6-H), 2.09 (s, 3H, Me), 1.99 EMI36.3 (ddd, J= 14.5, 11.5, 3.5 Hz, 1H, 6-H), 1.81 (br s, 1H, OH), 1.65 (s, 3H, Me), 1.24 (s, 3H, Me), 1.16 (s, 3H, Me), 0.91 (t, J= 8.0 Hz, <Desc/Clms Page number 37> EMI37.1 9H, OSi(CH?C-I3)3), 0.60-0.46 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA / CsI) m / e 753.1530, M + Cs+ calcd for C31H4409SSi 753.1530. Preparation of 2-pyridinyl-C-2 ester derivatives (Alcohol 15, 16 and triol 6) EMI37.2 Alcohol 15 and 16, and triol 6. A solution of carbonate 7 (62.6 mg, 0.108 mmol) in tetrahydrofuran (5.4 mL) at -78 C was treated with 2-lithiopyridine (1.15 mL of a 0.44 M solution in diethylether-pentane 1 : 1, 0.506 mmol, <Desc/Clms Page number 38> prepared from 2-bromopyridine and t-Butyllithium; methodology from Malmberg, H. ; Nilsson, M. Tetrahedron, 1986,42, 3981) and stirred for 1.3 hour The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NH4Cl (5 mL), the organic layer was separated, and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 70 # 100% ethylacetate in petroleum ether) to give 6 (16.3 mg, 27%), 15 (28.0 mg, 39%), and 16 (8.4 mg, 13%) as amorphous solids. Physical Data for Alcohol 15. Rf = 0.60 (silica, ethylacetate); lH NMR (500 MHz, CDC13) 8 8.77 (ddd, J= 4.5, 1.7, 1.0 Hz, 1H, pyridine), 8.05 (br d, J = 7.5 Hz, 1H, pyridine). 7.89 (ddd, J= 7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.53 (ddd, J= 7.5, 4.5, 1.0 Hz, 1H, pyridine), 5.61 (dd,J= 6.5, 1.0 Hz, 1H, 2-H), 5.33 (d, J = 2.5 Hz, 1H, 10-H), 4.92 (dd, J = 9.5, 2.0 Hz, 1H, 5-H), 4.39 (dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.36 (d, J = 9.0 Hz, 1H, 20-H), 4.33 (d, J = 2.5 Hz, 1H, 10-OH), 4.28 (d, J = 9.0 Hz, 1H, 20-H), 3.96 (d, J= 6.5 Hz, 1H, 3-H), 2.98 (d, J= 20.0 Hz, 1H, 14-H), 2.71 (dd, J= 20.0,1.0 Hz, 1H, 14-H), 2.50 (s, 1H, OH), 2.48 (ddd, J= 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.15 (s, 3H, Me). 2.11 (s, 3H, Me), 1.90 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.76 (s, 3H, Me), 1.24 (s, 3H, Me), 1.16 (s, 3H, Me), 0.93 (t, J = 8.0 Hz, 9H, EMI38.1 OSi(CH2CH3)3), 0.63-0.47 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA / Csl) m / e 790.2060, M + Cs- calcd for C3-J.H-J.701ONSi 790.2024. <Desc/Clms Page number 39> Physical Data for Alcohol 16. Rf = 0.45 (silica, ethylacetate); IR (film) vmax 3435, 2954, 2879, 1732,1674, 1589, 1362, 1305, 1241, 1116, 998, 829, 741 cm-1; 1H NMR (500 MHz, CDC13) # 8.73 (br d, J= 4.5 Hz, 1H, pyridine), 8.15 (br d, J= 7.5 Hz, 1H, pyridine), 7.90 (ddd, J= 7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.56 (ddd, J= 7.5, 4.5, 1.0 Hz, 1H, pyridine), 5.53 (dd, J= 7.5,1.0, 1H, 2-H), 5.30 (d, J= 2.5 Hz, 1H, 10-H), 4.84 (dd, J= 9.5, 3.0 Hz, 1H, 5-H), 4.81 (br s, 1H, OH), 4.31 (d, J= 2.5 Hz, 1H, 10-OH), 4.25 (s, 2H, 20-CH2), 3.97 (dd, J= 11.5, 6.5 Hz, 1H, 7-H), 3.31 (d, J= 19.5 Hz, 1H, 14-H), 3.23 (d, J= 7.5 Hz, 1H, 3-H), 2.57 (br d, J= 19.5 Hz, 1H, 14-H), 2.43 (ddd, J= 14.5,9.5, 6.5 Hz, 1H, 6-H), 2.11 (s, 3H, Me), 1.95 (ddd, J= 14.5, 11.5, 3.0 Hz, 1H, 6-H), 1.92 (br s; 1H, OH), 1.70 (s, 3H, Me), 1.24 (s, 3H, Me), 1.17 (s, 3H, Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.60-0.46 (band, 6H, OSi(CH?CH3)3). Physical Data for Triol 6. Rf= 0.24 (silica, 50% EMI39.1 ethylacetate in hexanes); IR (thin film) va; 3414, 297, 2881, 1727, 1664, 1370 cm-1; 1H NMR (500 MHz, CDC1;) 5 .23 (d, J= 9.5 Hz, 1 H, 10-H), 4.89 (d, J= 9.5 Hz, 1 H, 5-H), 4.63 (d, J= 9.5 Hz, 1 H, 20-H), 4.56 (d, J= 9.5 Hz, 1 H, 20-H), 4.32 (dd, J= 11.0, 7.0 Hz, 1 H, 7-H), 4.28 (d, J= 2.5 Hz, 1 H,10-OH), 3.89 (dd, J= 6.5, 4.0 Hz, 1 H, 2-H), 3.57 (d, J = 6.5 Hz, 1 H, 3-H), 2.78 (d, J= 19.5 Hz, 1 H, 14-H), 2.58 (d, 4.0 Hz, 1 H, 2-OH), 2.52 (d, J= 19.5 Hz, 1 H, 14-H), 2.49-2.42 (m, 1 H, 6-H), 2.03 (s, 3 H, Me), 1.92- 1.84 (m, 1 H, 6-H), 1.68 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.04 (s, 3 H, Me), 0.90 (t, J= 8.0 Hz, 9 H, Si(CH2CH3)3), 0.60-0.40 (band, 6 EMI39.2 H,Si(CH2CH3)3); 13C Ni4R (125 l\1Hz, CDC13) 8 208.9, 198.3, 170.1, <Desc/Clms Page number 40> 156.7, 138.8, 83.8, 81.2, 77.6, 75.7, 72.8, 72.5, 58.8, 45.8, 43.1, 42.8, 37.3, 32.7, 21.6, 17.5, 13.6,9.7, 6.7, 5.1; FAB HRMS (NBA NaI) m / e 575.2648, M + Na+'calcd for C28H4409Si 575.2652. Preparation of 3-pyridinyl-C-2 ester derivative (Alcohol 17) EMI40.1 Alcohol 17. To a solution of 3-lithiopyridine (1.15 mmol) in tetrahydrofuran (7 mL), prepared from 3bromopyridine (Aldrich Chemical Company Inc. ) and nButyllithium (Aldrich Chemical Company Inc.) at -100 C EMI40.2 (methodology from Parham, h%.E.; Piccirilli, R. M. J. Org. Chem. 1977, 42, 257), was added a solution of carbonate 7 (133.1 mg, 0.230 mmol) in tetrahydrofuran (2 mL) via cannula. The resulting solution was stirred for 1 h, allowed to warm to -78 C, stirred for 1 h, and poured into a mixture of ethylacetate (10 mL) and aqueous NH4Cl (10 mL). The organic layer was separated and the aqueous layer was extracted with <Desc/Clms Page number 41> ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (10, mL), dried (MgS04), concentrated, and purified by , flash chromatography (silica, 70 # 95% ethylacetate in petroleum ether) to give 7 (64.8 mg, 49%) and 17 (43.9 mg, 57% based on 51% conversion) as an amorphous solid. Physical Data for Alcohol 17. Rf= 0.56 (silica, ethylacetate) ; IR (film) #max 3435, 2956, 2882, 1731,1671, 1592,1366, 1280,1240, 1109, 991,824, 739 cm-1; 1H NMR (500 MHz, CDC13) # 9.24 (br s, 1H, pyridine), 8.81 (d, J= 1.0,4.5 Hz, 1H, pyridine), 8.30 (ddd, J= 8.0, 2.0,2.0 Hz, 1H, pyridine), 7.44 (dd, J= 8.0, 4.5 Hz, 1H, pyridine), 5.66 (d, J= 6.5 Hz, 1H, 2H), 5.32 (s, 1H, 10-H), 4.92, (dd, J = 9.5,2.0 Hz, 1H,] 5-H), 4.38 (dd, J= 10.5,6.5 Hz, 1H, 7-H), 4.32 (br s, 1H, OH), 4.30 (d, J= 8.5 Hz, 1H, 20-H), 4.13 (d, J= 8.5 Hz, 1H, 20-H), 3.96 (d, J= 6.5 Hz, 1H, 3-H), 2.92 (d, J= 19.5 Hz, 1H. 14-H), 2.66 (d, J= 19.5 Hz, 1H, 14-H), 2.48 (ddd, J= 15.5, 9.5, 6.5 Hz, 1H, 6-H), 2.18 (s, 3H, Me), 2.10 (s, 3H, Me), 2.03 (s, 1H, OH), 1.89 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.72 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.62-0.48 (band, 6H, OSi(CH?CH3)3); FAB HRMS (NBA / CSI) m / e 790.2030, M + Cs+ EMI41.1 calcd for C3.H.0lpNSi790.2024. <Desc/Clms Page number 42> Preparation of 4-N,N-dimethylaniline-C-2 ester derivative (Alcohol 18) EMI42.1 Alcohol 18. A solution of carbonate 7 ( 150 mg, 0.259 mmol) in tetrahydrofuran ( 20 mL) at -78 C was treated EMI42.2 with 4-lithio-N,1' dimethylanilirie (6.5 mL of a 0.39 M solution in diethylether : pentane (3 : 1), 2.54 mmol, prepared from 4bromo-N,N-dimethylaniline and t-Butyllithium; methodology from Jones, F. N.; Hauser, C.R. J. Org. Chem. 1962, 27, 4389) and stirred for 15 minutesutes The reaction mixture was poured into a mixture of CH2C12 (35 mL) and aqueous NH4CI (20 mL), the organic layer was separated, and the aqueous layer was extracted with CH2C12 (2 x 20 mL). The combined organic layer was washed with brine (20 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10 - 35% ethylacetate in petroleum ether) to give 18 (55.0 mg, 30%) as an amorphous solid. Physical Data for Alcohol 18. Rf = 0.26 (silica, 35% ethylacetate in hexanes) ; IR (film) vmax 3414, 2924, 1706, 1669, 1605, 1530, 1094; lH NMR (500 MHz, CDC13) # 7.90 (d, J= <Desc/Clms Page number 43> 9.0 Hz, 2H, Ar), 6.64 (d, J = 9.0 Hz, 2H, Ar), 5.60 (br d, J = 7.0 Hz, 1H, 2-H), 5.29 (d, J = 2.5 Hz, 1H, 10-H), 4.89 (br d, J = 9.5 Hz, EMI43.1 1H, 5-H), 4.37 (d, J= 8.5 Hz, 1H, 20-H), 4.36 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.31 (d, J= 2.5 Hz, 1H, 10-OH), 4.13 (br d, J= 8.5 Hz, 1H, 20-H), 3.90 (d, J= 7.0 Hz, 1H, 3-H), 3.05 (s, 6H, NMe2), 2.93 (s, 1H, OH), 2.90 (d, J= 20.0 Hz, 1H, 14-H), 2.61 (br d, J= 20.0 Hz, 1H, 14-H), 2.49-2.40 (m, 1H, 6-H), 2.16 (s, 3H, Me), 2.08 (s, 3H, Me), 1.90-1.83 (m, 1H, 6-H), 1.69 (s, 3H, Me), 1.20 (s, 3H, Me), 1.13 (s, 3H, Me), 0.90 (t, J = 8.0 Hz, 9H, OSi(CH2CH3)3), 0.56-0.49 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ NaI) m / e 722.3354, M + Na+ calcd for C73H53O10NSi 722.3336. Preparation of 1-naphthalene-C-2 ester derivative (Alcohol 19) EMI43.2 Alcohol 19. A solution of carbonate 7 (47 mg, 0.0812 mmol) in tetrahydrofuran (2 mL) at -78 C was treated with 1lithionaphthalene (6.3 mL of a 0.32 M solution in diethylether, <Desc/Clms Page number 44> 2.03 mmol, prepared from 1-bromonaphthalene from Aldrich Chemical Company Inc. and tButyllithium; methodology from Gilman, H. ; Moore, F.W. J. Am. Chem. Soc. 1940, 62, 1843) and stirred for 5 minutesutes The reaction mixture was poured into a mixture of CH2C12 (15 mL) and aqueous NH4Cl (20 mL), the organic layer was separated, and the aqueous layer was extracted with CH2C12 (2 x 10 mL). The combined organic layer was washed with brine (10 mL), dried (MgS04) and concentrated to give alcohol 19 which was taken into the next step without further purification. Physical Data for Alcohol 19. Rf= 0.27 (20% ethylacetate in petroleum ether); IR (film) #max 3442, 2954, 2882,1724,1671,1461,1362,1279,1228,1195,1092,987, EMI44.1 826, 736 cm-1; 11H NMR (500 MHz, CDC13) 8 8.66 (s, 1H, naphthalene), 8.07 (dd, J= 9.0, 2.0 Hz, 1H, naphthalene), 7.97- 7.89 (m, 3H, naphthalene), 7.68-7.57 (m, 2H, naphthalene), 5.71 (br d, J = 6.5 Hz, 1H, 2-H), 5.35 (d, J = 2.5 Hz, 1H, 10-H), 4.94 (br d, J= 8.0 Hz, 1H, 5-H), 4.41 (dd, J= 11.0, 7.0 Hz, 1H, 7H), 4.37 (d, J= 8.5 Hz, 1H, 20-H), 4.35 (d, J= 2.0 Hz, 1H, 10-OH), 4.18 (d, J= 8.5 Hz, 1H, 20-H), 4.00 (d, J= 6.5 Hz, 1H, 3-H), 3.02 (d, J= 19.5 Hz, 1H, 14-H), 2.69 (d, J= 19.5 Hz, 1H, 14-H), 2.54- 2.45 (m, 1H, 6-H), 2.27 (s, 3H, Me), 2.13 (s, 3H, Me), 1.94-1.87 (m, 1H, 6-H), 1.86 (s, 1H, OH), 1.75 (s, 3H, Me), 1.25 (s, 3H, Me), 1.20 (s, 3H; Me), 0.94 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.63-0.49 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA) m / e 707.3270, M EMI44.2 + Hy calcd for C 39H]oojosi 707.3252. <Desc/Clms Page number 45> Preparation of phenylacetylide-C-2 ester derivative (Alcohol 20) EMI45.1 Alcohol 20. A solution of carbonate 7 (5.0 mg, 0.00864 mmol) in tetrahydrofuran (0.5 mL) at -78 C was treated with lithium phenylacetylide from Aldrich Chemical Company Inc. (0.13 mL of a 1.0 M solution in tetrahydrofuran, 0.13 mmol) and stirred for 0.5 hour The reaction mixture was treated with aqueous NH4CI (0.5 mL), allowed to warm to 25 C, and diluted with H20 (5 mL) and diethylether (5 mL) . The organic layer was separated, dried, and concentrated to give a 9 : 1 mixture of carbonate 7 and alcohol 20 (5.0 mg, 95%) as a film. Physical Data for Alcohol 20. Rf= 0.59 (50% ethylacetate in hexanes); 1H NMR (300 MHz, CDCl3) 8 7.63-7.57 (m, 2 H, Ar), 7.53-7.27 (m, 3 H, Ar), 5.43 (d, J= 6.5 Hz, 1H, 2-H), 5.28 (d, J = 2.5 Hz, 1H, 10-H), 4.90 (br d, J = 7.5 Hz, 1H, 5-H), 4.67 (d, J = 8.5 Hz, 1H, 20-H), 4.44 (d, J = 8.5 Hz, 1H, 20-H), 4.37-4.30 (m, 1 H, 7-H), 4.28 (d, J= 2.5 Hz. 1H, 10-OH). 3.88 (d. J= 6.5 Hz, 1H, 3-H), 2.85 (d, J= 20.2 Hz, 1H, 14-H), 2.63 (d, J= 20.2 Hz, 1H, 14-H), 2.55-2.47 (m, 1 H, 6-H), 2.11 (s, 3 H, OAc), <Desc/Clms Page number 46> 2.08 (s, 3 H, 18-Me), 1.94-1.85 (m, 1 H, 6-H), 1.67 (s, 3 H, Me), 1.41 (s, 3 H, Me), 1.21 (s, 3 H, Me), 0.91 (t, J = 8.0 Hz, 9H, EMI46.1 OSi(CH?C-I3)3}, 0.59-0.42 (band, 6H, OSi(CH2CH3)3). Preparation of Hydroxycarbamate-C-2 ester derivative (Alcohol 21) EMI46.2 Alcohol 21. A solution of carbonate 7 (5 .0 mg, 0.00864 mmol) in MeOH (0.5 mL) at 25 C was treated with n-Butyl-NH2 from Aldrich Chemical Company Inc. (0.05 mL, 0.506 mmol) and stirred for 10 minutes. The reaction mixture was concentrated and purified by flash chromatography (silica, 30 # 50% ethylacetate in petroleum ether) to give 21 (5.2 mg, 92%) as an amorphous solid. Physical Data for Alcohol 21. Rf = 0.13 (silica, 30% ethylacetate in petroleum ether); IR (film) #max 3434, 2957, 2881, 1711, 1671, 1368, 1243, 1108, 987, 829 cm-1; 1H NMR (500 MHz, CDC13) # 5.27 (d, J= 2.0 Hz, 1H, 10-H). 5.23 (d, J= 6.5 Hz, 1H, 2-H), 4.91 (br d, J= 8.0 Hz, 1H, 5-H), 4.79 (t, J= 6.0 Hz, 1H, NH), 4.47 (d, J= 8.5 Hz, 1H, 20-H), 4.34 (dd, J= 11.0, 7.0 Hz, <Desc/Clms Page number 47> 1H, 7-H), 4.30 (d, J= 2.5 Hz, 1H, 10-OH), 4.28 (d, J= 8.5 Hz, 1H, 20-H), 3.78 (d, J = 6.5 Hz, 1H, 3-H),, 3.29-3.12 (m, 2H, NHCH2), 2.70 (d, J= 20.0 Hz, 1H, 14-H), 2.60 (d, J= 20.0 Hz, 1H, 14-H), 2.51-2.42 (m, 1H, 6-H), 2.24 (s, 1H, OH), 2.06 (s, 3H, Me), 2.05 (s, 3H, Me), 1.94-1.86 (m, 1H, 6-H), 1.69 (s, 3H, Me), 1.55-1.46 (m, 2H, NHCH2CH2), 1.40-1.30 (m, 2H, NHCH2CH2CH2), 1.21 (s, 3H, Me), 1.09 (s, 3H, Me), 0.95-0.80 (m, 12H, CH3 of Bu, OSi(CH2CH3)3), 0.61-0.47 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ NaI) m / e 674.3336, M + Na- calcd for C33HS3010NSi 674.3336. EMI47.1 Preparation of NN methyl-phenyl-hydroycarbamate- C-2 ester derivative (Alcohol 22)' EMI47.2 Alcohol 22. A solution of carbonate 7 (5.0 mg, 0.00864 mmol) in tetrahydrofuran (0.5 mL) at -78 C was treated with LiNMePh (0.2 mL of a 0.47 M solution in diethylether, 0.094 mmol, prepared from N-methylaniline (Aldrich) and nButyllithium) and stirred for 1.25 hour The reaction mixture was poured into a mixture of diethylether (5 mL) and aqueous <Desc/Clms Page number 48> NH4Cl (5 mL), the organic layer was separated, and the aqueous layer was extracted with diethylether (2 x 5 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 15 # 35% ethylacetate in hexanes) to give 7 (2.5 mg, 50%) and 22 (2.8 mg, 93% based on 50% conversion) as an amorphous solid. Physical Data for Alcohol 22. Rf= 0.22 (silica 35% EMI48.1 ethylacetate in petroleum ether); ,1 H NMR ( 00 MHz, CDC13) 8 7.45-7.18 (band, 5H), 5.25 (br d, J= 6.5 Hz, 1H, 2-H), 5.20 (d, J = 2.5 Hz, 1H, 10-H),4.70 (br d, J= 8.0 Hz, 1H, 5-H), 4.26 (d, J= 2.5 Hz, 1H, 10-OH), 4.22 (dd, J= 10.5, 6.5 Hz. 1H, 7-H), 4.19 (d, J = 8.5 Hz, 1H, 20-H), 4.16 (d, J= 8.5 Hz, 1H, 20-H), 3.58 (d, J= 7.0 Hz, 1H, 3-H), 3.27 (s, 3H, MeN), 2.52 (d, J= 20.0 Hz, 1H, 14H), 2.35 (d, J= 20.0 Hz, 1H, 14-H), 2.40-2.31 (m, 1H, 6-H), 2.03 (s, 1H, OH), 1.97 (s, 3H, Me), 1.85-1.76 (m, 1H, 6-H), 1.66 (s, 3H, Me), 1.57 (s, 3H, Me), 1.18 (s, 3H, Me), 1.08 (s, 3H, Me), 0.87 (t, J = 8.0 Hz, 9H, OSi(CH2CH3)3), 0.55-0.43 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA) m/ e 686.3358, M + H- calcd for C36H51O10NSi 686.3361. <Desc/Clms Page number 49> Preparation of Thioether-C-2 ester derivative (Alcohol 23) EMI49.1 Alcohol 23. A solution of vinyl ester 9 (55.6 mg, 0.0916 mmol) and 4-dimethylaminutesopyridine from Aldrich Chemical Company Inc. (DMAP, 1.8 mg, 0.0147 mmol) in CH2C12 (4.3 mL) at 25 C was treated with PhSH from Aldrich Chemical Company Inc. (0.030 mL, 0.292 mmol) and stirred for 1.5 hour The reaction mixture was concentrated and purified by flash chromatography (silica, 30% ethylacetate in petroleum ether) to give 23 (58.1, 88%) as a white solid. Physical Data for Alcohol 23. Rf= 0.37 (silica, 30% ethylacetate in hexanes), 0.34 (10% ethylacetate in PhH, 2 elutions) ; IR (film) vmax 3441, 3057, 2956, 2883, 1732, 1672, 1600, 1367, 1238, 1111, 988, 825, 739 cm -1; 1H NMR (500 MHz, CDC13) 8 7.39-7.24 (band, 5H), 5.44 (d, J= 6.5 Hz, 1H, 2-H), 5.28 (d, J= 2.5 Hz, 1H, 10-H), 4.90 (dd, J= 9.5,2.0 Hz, 1H, 5-H), 4.38 (d, J= 8.0 Hz, 1H, 20-H), 4.33 (dd, J= 10.5, 6.5Hz, 1H. 7-H), 4.29 (d, J = 2.5 Hz, 1H, 10-OH), 4.18 (d, J = 8.0 Hz, 1H, 20-H), 3.83 (d, J= 6.5 Hz, 1H, 3-H), 3.24-3.13 (m, 2H, CH2SPh), 2.76 (d, <Desc/Clms Page number 50> J = 19.5 Hz, 1H, 14-H), 2.72-2.58 (m, 3H, 14-H, CH2CH2SPh), 2.47 (ddd, J= 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.39 (s, 1H, OH), 2.07 (s, 3H, Me), 2.05 (s, 3H, Me), 1.89 (ddd, J= 14.5,10.5, 2.0 Hz, 1H, 6-H), 1.68 (s, 3H, Me), 1.23 (s, 3H, Me), 1.12 (s, 3H, Me), 0.92 (t, J = 8.0 Hz, 9H, OSi(CH2CH3)3), 0.61-0.47 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m / e 849.2085, M + Cs+ calcd for C37H52O10SSi 849.2105. Preparation of intermediates 25-27 and 2-furanyl-C- 2-taxoid (28) EMI50.1 Acetate 25. A solution of alcohol 11 and @- dimethylaminopyridine (DMAP, 100 mg, 0.819 mmol) in CH2C12 (3 mL) at 25 C was treated with acetic anhydride (0.50 mL, <Desc/Clms Page number 51> 5.30 mmol) and stirred for 3 h. The reaction mixture was diluted with CH2C12 (5 mL), treated with aqueous NaHC03 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by preparative TLC (silica, 10% ethylacetate in benzene, 3 elutions) to give 25 (36 mg, 66% from carbonate 7) as a white foam. Physical Data for Acetate 25. Rf = 0.38 (20% ethylacetate in petroleum ether); IR (film) vmax 3509, 2956, 2881,1727,1674,1469,1371,1299,1227,1108,746 cm-1; 1H NMR (500 MHz, CDC13) 8 7.65 (br s, 1H, furan), 7.24 ( br d, J = 3.0 Hz, 1H, furan), 6.58-6.54 (m, 2H, 10-H, furan), 5.59 (d, J= 6.5 Hz, 1H, 2-H), 4.92 (br d, J = 7.5 Hz, 1H, 5-H), 4.46 (dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.42 (d, J= 8.5 Hz, 1H, 20-H), 4.16 (d, J= 8.5 Hz, 1H, 20-H), 3.87 (d, J = 6.5 Hz, 1H, 3-H), 2.89 (d, J= 20.0 Hz, 1H, 14-H), 2.63 (d, J= 20.0 Hz, 1H, 14-H), 2.59-2.48 (m, 1H, 6-H), 2.22 (s, 3H, Me), 2.17 (s, 3H, Me), 2.14 (s, 3H, Me), 1.90- 1.83 (m, 1H, 6-H), 1.65 (s, 3H, Me), 1.25 (s, 3H, Me), 1.18 (s, 3H, Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.64-0.52 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m / e 821.1966, M + Cs+ calcd for C35H48O12Si 821.1969. Alcohol 26. A solution of enone 25 (36 mg, 0.0523 mmol) in MeOH (3 mL) containing two drops of CH3COOH at 0 C was treated with NaBH4 (200 mg, 5.29 mmol, added by <Desc/Clms Page number 52> portions) and stirred for 6 h. The reaction mixture was diluted with CH2C12 (10 mL), treated with aqueous NH4CI (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2C12 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by preparative TLC (silica, 50% ethylacetate in petroleum ether) to give 26 (30 mg, 83% ) as an amorphous solid. Physical Data for Alcohol 26. Rf = 0.42 (silica, 50% ethylacetate in petroleum ether); 1H NMR (300 MHz, CDC13) 8 7.62 (br s, 1H, furan), 7.25 (d, J= 3.5 Hz, 1H, furan), 6.58 (d, J= 3.5 Hz, 1H, furan), 6.43 (s, 1 H, 10-H), 5.51 (d, J= 7.0 Hz, 1H, 2H), 4.96 (d, J = 7.5 Hz, 1H, 5-H), 4.85-4.79 (m, 1 H, 13-H), 4.48 (dd, J = 10.5, 7.5 Hz, 1H, 7-H), 4.38 (d, J = 8.0 Hz, 1H, 20-H), 4.15 (d, J= 8.0 Hz, 1H, 20-H), 3.82 (d, J= 7.0 Hz, 1H, 3-H), 2.61- 2.48 (m, 2 H, 6-H and 14-H), 2.28 (s, 3 H, OAc), 2.20-2.10 (m, 1 H, 14-H), 2.18 (s, 6 H, OAc and 18-Me), 1.98-1.80 (m, 1 H, 6-H), 1.18 (s, 3 H, 16-Me), 1.04 (s, 3 H, 17-Me), 0.90 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.65-0.50 (band, 6H, OSi(CH2CH3)3). DiTES taxoid 27. To a solution of alcohol 26 (30.0 mg, 0.0434 mmol, previously azeotroped twice with benzene) and P-lactam 24 (28.0 mg, 0.0734 mmol, previously azeotroped twice with benzene) in THF (2 mL), prepared from the OjimaHolton protocol (Holton, R. A. Chem .4bstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; <Desc/Clms Page number 53> Zucco, M. ; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, EMI53.1 48, 6985-7012), at 0 C was added NaN(SiMe3)2 (0.130 mL of a 1.0 M solution in THF, 0.130mmol) dropwise. The resulting solution was stirred for 5 min and poured into a mixture of CH2Cl2 (10 mL) and aqueous NH4CI (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2C12 (2 x 5 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by preparative TLC (silica,' 60% ethylacetate in petroleum ether) to give 27 (12 mg, 26%) as an amorphous solid which was taken directly into the next step. Taxoid 2 8. A solution of silyl ether 27 (6 mg, 0.00560 EMI53.2 mmol) in THF (1 mL) at 25 C was'-treated with HF. pyridine (1 mL) and stirred for 1 h..The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (10 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 60% ethylacetate in petroleum ether) to give 28 (3 mg, 64%) as a colorless film. Physical Data for Taxoid 28. Rf= 0.1 (50% ethylacetate in petroleum ether); IR (film) #max 3383, 2933, 2898, 1729, 1649, 1519, 1242, 1110, 1071 cm-1; 1H NMR (500 MHz, CDCl3) # 7.77-7.73 (m, 2H), 7.68-7.66 (m, 1H, furan), 7.55- 7.33 (band, 9H), 6.98 (d, J= 9.0 Hz, 1H, NH), 6.58 (dd, J= 3.5, <Desc/Clms Page number 54> 1.5 Hz, 1H, furan), 6.27-6.21 (m, 2H,,10-H, 13-H), 5.80 (dd, J= 9.0, 2.0 Hz, 1H, 3'-H), 5.57 (d, J= 7.0 Hz, 1H, 2-H), 4.96 (dd, J= EMI54.1 10.0, 2.0 Hz, 1H, 5-H), ,4.80 (d, J= 2.OHz, 1H, 2'-H), 4.43-4.37 (m, 2H, 7-H, 20-H), 4.24 (d, J= 8.5 Hz, 1H, 20-H), 3.77 (d, J= 7.0 Hz,l H, 3-H), 2.60-2.52 (m, 1H, 6-H), 2.47 (d, J= 4.0 Hz, 1H, OH), 2.38 (s, 3H, Me), 2.35-2.21 (m, 2H, 14-CH2), 2.25 (s, 3H, Me), 1.94-1.86 (m, 1H, 6-H), 1.81 (br s, 3 H, Me), 1.76 (s, 1H, OH), 1.68 (s, 3H, Me), 1.25 (s, 3H, Me), 1.13 (s, 3H, Me). Preparation of 2-thiophenyl-C-2 taxol (32) EMI54.2 Acetate 29. A solution of alcohol 12 (36.0 mg, 0.0543 mmol) and 4-dimethylaminopyridine (DNIAP, 33.0 mg, 0.270 <Desc/Clms Page number 55> mmol) in CH2C12 (3.0 mL) at 25 C was treated with acetic anhydride (0.50 mL, 5.30 mmol) and stirred for 1 h. The reaction mixture was diluted with CH2C12 (10 mL), treated with aqueous NaHC03 (7 mL), and stirred vigorously for 0.5 h. The organic, layer was separated and the aqueous layer was extracted with CH2C12 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 10 # 35% ethylacetate in hexanes) to give 29 (29.5 mg, 77%) as an amorphous solid. Physical Data for Acetate 29. Rf = 0.56 (silica, 50% ethylacetate in petroleum ether); IR (film) #max 3457, 2956, EMI55.1 1712, 1669, 1525, 1413, 1376, 1264, 1227, 1073; 1H NMR (500 MHz, CDCl3) 8 7.84 (dd, J= 4.0, 1.5 Hz, 1H, thiophene), 7.63 (dd, J = 5.0, 1.5 Hz, 1H, thiophene), 7.13 (dd, J = 5.0, 4.0 Hz, 1H, thiophene), 6.56 (s, 1H, 10-H), 5.58 (br d, J= 6.5 Hz, 1H, 2-H), 4.90 (br d, J= 8.0 Hz, 1H, 5-H), 4.44 (dd, J= 10.5, 7.0 Hz, 1H, 7H), 4.42 (d, J= 8.5 Hz, 1H, 20-H), 4.18 (d, J= 8.5 Hz, 1H, 20-H), 3.85 (d, J= 6.5 Hz, 1H, 3-H), 2.91 (d, J= 19.5 Hz, 1H, 14-H), 2.64 (dd, J= 19.5,1.0 Hz, 1H, 14-H), 2.55-2.48 (m, 1H, 6-H), 2.20 (s, 3H, Me), 2.15 (s, 3H, Me), 2.14 (s, 3H, Me), 1.89-1.82 (m, 1H, 6H), 1.65 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), 0.88 (t, J = 8.0 Hz, 9H, OSi(CH2CH3)3), 0.59-0.53 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) in / e 837.1736, M + Cs- calcd for EMI55.2 C 3 5 fi480 1 1 SSi 837.1741. Alcohol 30. A solution of enone 29 (29.0 mg, 0.0411 <Desc/Clms Page number 56> mmol) in MeOH (5 mL) at 0 C was treated with NaBH4 (30.2 mg, 0.80 mmol, added by portions) and stirred for 2.5 h. The reaction mixture was diluted with CH2Cl2 (15 mL), treated with aqueous NH4Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 25 -- 50% ethylacetate in petroleum ether) to give 29 (4.0 mg, 14%) and 30 ( 14.7 mg, 59% based on 86% conversion) as an amorphous solid. Physical Data for Alcohol 30. Rf= 0.34 (silica, 50% ethylacetate in petroleum ether); IR (film) #max 3478, 2946, 2892, 1717, 1520, 1365, 1238, 1083; 1H NMR (500 MHz, CDCI3) 8 7.85 (dd, J= 3.5, 1.5 Hz, 1H, thiophene), 7.61 (dd, J= 5.0, 1.5 Hz, 1H, thiophene), 7.12 (dd, J= 5.0, 3.5 Hz, 1H, thiophene), 6.43 (s, 1H, 10-H), 5.51 (d, J= 7.0 Hz, 1H, 2-H), 4.94 (br d, J = 7.5 Hz, 1H, 5-H), 4.83-4.77 (m, 1H, 13-H), 4.45 (dd, J= 10.5, 7.5 Hz, 1H, 7-H), 4.41 (d, J = 8.0 Hz, 1H, 20-H), 4.19 (br d, J = 8.0 Hz, 1H, 20-H), 3.82 (d, J= 7.0 Hz, 1H, 3-H), 2.55-2.48 (m, 1H, 6-H), 2.24 (s, 3H, Me), 2.26-2.21 (m, 2H, 14-CH2), 2.16 (d, J= 1.0 Hz, 3H, 18-Me), 2.15 (s, 3H, Me), 2.00 (d, J= 5.0 Hz, 1H, OH), 1.90-1.82 (m, 1H, 6-H), 1.66 (s, 3H, Me), 1.58 (s, 1H, OH), 1.15 (s, 3H, Me), 1.02 (s, 3H, Me), 0.90 (t, J= 8.0 Hz, 9H, OSiCH2CH3)3), 0.59-0.55 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA / CsI) m / e 839.1893, M + Cs- calcd for C35H50O11SSi 839.1897. DiTES taxoid 31. To a solution of alcohol 30 (14.5 mg, <Desc/Clms Page number 57> 0.0205 mmol, previously azeotroped :twice with benzene) and P-lactam 24 (16.0 mg, 0.0420 mmol, previously azeotroped twice with benzene) in THF (1.0 mL), prepared from the OjimaHolton protocol (Holton, R.A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M. ; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), at -78 C was added NaN(SiMe3)2 (0.051 mL of a 1.0 M solution in THF, 0.051 mmol) dropwise. The resulting solution was stirred for 0.5 h and poured into a mixture of diethylether (10 mL) and aqueous NH4Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with diethylether (2 x 5 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatog.raphy (silica, 10 # 35% ethylacetate in hexanes) followed by preparative TLC (silica, 15% ethylacetate in benzene) to give 30 (3.0 mg, 21%) and 31 (7.6 mg, 43% based on 79% conversion) as a white solid. Physical Data for DiTES taxoid 31. Rf = 0.48 (silica, 50% ethylacetate in hexanes); IR (film) #max 3382, 2913, 2850, 1722, 1653;1461,1243,1083, 1014; 1H NMR (500 MHz, CDCl3) # 7.90 (br d, J = 4.0 Hz, 1H, thiophene), 7.74 (d, J= 8.0 Hz, 2H, NBz), 7.62 (br d, J =5.0 Hz, 1H, thiophene), 7.48 (t, J= 7.0 Hz, 1H, Ar), 7.42-7.28 (band, 7H, Ar), 7.14 (dd, J= 5.0, 4.0 Hz, 1H, thiophene), 7.10 (d, J= 9.0 Hz. 1H, NH), 6.42 (s, 1H, 10-H), 6.20 (br t, J = 9.0 Hz, 1H, 13-H), 5.65 (br d, J= 9.0 Hz, 1H, 3'-H), 5.57 (d, J=7.0 Hz, 1H, 2-H), 4.94 (br d, J = 8.5 Hz, 1H, 5-H), 4.67 (d, J <Desc/Clms Page number 58> = 1.5 Hz, 1H, 2'-H), 4.44 (dd, J= 11.0, 6.5 Hz, 1H, 7-H), 4.43 (d, J = 8.5 Hz, 1H, 20-H), 4.26 (d, J= 8.5 Hz, 1H, 20-H), 3.77 (d, J= 7.0 Hz, 1H, 3-H), 2.51 (s, 3H, Me), 2.54-2.47 (m, 1H, 6-H), 2.34 (dd, J = 15.0, 9.5 Hz, 1H, 14-H), 2.15 (s, 3H, Me), 2.10 (dd, J= 15.0, 9.0, 1H, 14-H), 1.99 (s, 3H, Me), 1.93-1.86 (m, 1H, 6-H), 1.72 (s, 1H, OH), 1.68 (s, 3H, Me), 1.18 (s, 3H, Me), 1.16 (s, 3H, Me), 0.90 (t, J= 8.0 Hz, 9H, Si(CH2CH3)3), 0.79 (t, J= 8.0 Hz, 9H, EMI58.1 Si(CHCH3)3), 0.57-0.55 (band, 6H, Si(CH?CH3)3), 0.45-0.40 (band, 6H, Si(CH2CH3)3); FAB ,HRMS (NBA/ CsI) m / e 1220.3685 M + Cs+ calcd for C57H77O14NSSi2 1220.3658. Taxoid 32. A solution of silyl ether 31 (7.5 mg, 0.00689 mmol) in THF (0.8 mL) at 25 C was treated with HF#pyridine (0.150 mL) and stirred for 1 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHC03 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate ( 2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 50 # 100% ethylacetate in petroleum ether) to give 32 (4.2 mg, 71%) as a colorless film. Physical Data for Taxoid 32. Rf = 0.44 (silica, 75% ethylacetate in petroleum ether) ; IR (film) #max 3417, 2929, EMI58.2 1716, 1649, I3 21, 1460, 1- 1 i , 13 6 S, 12- i , 1076; 1 H ?WR ( 300 MHz, CDC13) # 7.90 (dd, J= 4.0. 1.0 Hz, 1H, thiophene), 7.73 (d, J 7.0 Hz, 2H, NBz), 7.63 (dd, J = 5.0, 1.0 Hz, 1H, thiophene), <Desc/Clms Page number 59> 7.51-7.32 (band, 8H, Ar), 7.14 (dd, J = 5.0,4.0 Hz, 1H, thiophene), 6.96 (d, J= 9.0 Hz, 1H, NH), 6.24 (s, 1H, 10-H), 6.19 (br t, J= 9.0 Hz, 1H, 13-H), 5.75 (dd, J= 9.0,2.5 Hz, 1H, 3'-H), 5.55 (d, J= 7.0 Hz, 1H, 2-H), 4.94 (br d, J= 8.0 Hz, 1H, 5-H), 4.76 (dd, J = 5.0, 2.5 Hz, 1H, 2'-H), 4.41 (d, J = 8.5 Hz, 1H, 20-H), 4.40-4.33 (m, 1H, 7-H), 4.24 (d, J= 8.5 Hz, 1H, 20-H), 3.73 (d, J = 7.0 Hz, 1H, 3-H), 3.52 (d, J= 5.0 Hz, 1H, 2'-OH), 2.58-2.49 (m, 1H, 6-H), 2.44 (d, J= 4.0 Hz, 1H, 7-OH), 2.35 (s, 3H, Me), 2.29 (d, J= 9.0 Hz, 2H, 14-CH?), 2.22 (s, 3H, Me), 1.91-1.83 (m, 1H, 6-H), 1.76 (s, 3H, Me), 1.66 (s, 3H, Me), 1.23 (s, 3H, Me), 1.10 (s, 3H, Me) ; FAB HRMS (NBA/ CsI) m / e 992.1252, M + Cs+ calcd for C45H49NO14S 992.1928. Preparation of 3-thiophenyl-C-2 taxol (36) EMI59.1 <Desc/Clms Page number 60> Acetate 3 3 . A solution of alcohol 13 (68.4 mg, 0.103 mmol) and 4-dimethylaminopyridine (DMAP, 37.8 mg, 0.309 mmol) in CH2Cl2 (4.4 mL) at 25 C was treated with acetic anhydride (0.370 mL, 3.92 mmol) and stirred for 2 h. The reaction mixture was diluted with CH2C12 (5 mL), treated with aqueous NaHC03 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in hexanes) to give 33 (66.0 mg, 91%) as an amorphous solid. Physical Data for Acetate 33. Rf= 0.48 (silica, 10% ethylacetate in benzene, 3 elutions); IR (film) #max 3518, 2956, EMI60.1 2881, 1727, 1676, 1520, 1460, 13 71, 1236, 1098, 985, 824, 744 cm-1; 1H NMR (500 MHz, CDC13) 5 8.19 (dd, J= 3.0, 1.1 Hz, 1H, thiophene), 7.55 (dd, J= 5.0, 1.1 Hz, 1H, thiophene), 7.38 (dd, J= 5.0,3.0 Hz, 1H, thiophene), 6.58 (s, 1H, 10-H), 5.61 (dd, J= 6.5, 0.7 Hz, 1H, 2-H), 4.92 (dd, J = 9.5, 2.0 Hz, 1H, 5-H), 4.47 (dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.38 (d, J = 8.5 Hz, 1H, 20-H), 4.14 (d, J= 8.5 Hz, 1H,' 20-H), 3.88 (d, J= 6.5 Hz, 1H, 3-H), 2.89 (d, J = 20 Hz, 1H, 14-H), 2.64 (br d, J = 20 Hz, 1H, 14-H), 2.54 (ddd, J= 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.23 (s, 3H, Me), 2.18 (s, 3H, Me), 2.17 (s, 3H, Me), 1.87 (ddd, J= 14.5, 10.5, 2.0, 1H, 6-H), 1.85 (s, 1H, OH), 1.66 (s, 3H, Me), 1.26 (s, 3H, Me), 1.19 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.65-0.54 (band, 6H, EMI60.2 OSi(CHCH3)3); FAB HRMS (NBA / Csl) m / e 837.1760, 1\1+Cs- calcd for C35H48O11SSi 837.1741. <Desc/Clms Page number 61> Alcohol 34. A solution,of enone 33 (57.3 mg, 0.0813 mmol) in MeOH-THF (5 : 1, 4.1 mL) at 0 C was treated with NaBH4 (69.1 mg, 1.83 mmol, added by portions) and stirred for 2.5 h. The reaction mixture was diluted with CH2Cl2 (10 mL), treated with aqueous NH4CI (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in hexanes) to give 33 (6.8 mg, 12%) and 34 (45.2 mg, 89% based on 88% conversion) as an amorphous solid. Physical Data for Alcohol 34. Rf= 0.48 (silica, 50% ethylacetate in hexanes); IR (film) #max 3520, 2953, 2881, 1719, 1520, 1370, 1238, 1100, 979, 823, 746 cm-1; 1H NMR (500 MHz, CDC13) 5 8.20 (dd, J= 3.0, 1.0 Hz, 1H, thiophene), 7.57 (dd, J= 5.0, 1.0 Hz, 1H, thiophene), 7.35 (dd, J= 5.0, 3.0 Hz, 1H, thiophene), 6.45 (s, 1H, 10-H), 5.54 (d, J= 7.0 Hz, 1H, 2-H), 4.96 (br d, J = 8.5 Hz, 1H, 5-H), 4.82 (br dd, J= 12.0, 8.0 Hz, 1H, 13H), 4.48 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.36 (d, J= 8.5 Hz, 1H, 20-H), 4.15 (d, J = 8.5 Hz, 1H, 20-H), 3.85 (d, J= 7.0 Hz, 1H, 3H), 2.53 (ddd, J= 14.5, 9.5, 6.5, 1H, 6-H), 2.27 (s, 3H, Me), 2.28- 2.21 (m, 2H, 14-CH2), 2.18 (s, 6H, Me, Me), 2.03 (s, 1H, OH), 1.87 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.67 (s, 3H, Me), 1.65 (s, 1H, OH), 1.18 (s, 3H, Me); 1.04 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, EMI61.1 OSi(CHCHy)3), 0.64-0.50 (band. 6H, OSi(CH2CH3L:;); FAB HRMS ( NBA / CsI) m / e 839.1908 lv1 + Cs+ calcd for C3SHS00l1SSi 839.1897. <Desc/Clms Page number 62> DiTES taxoid 3 5 . To a solution of alcohol 34 ( 19.5 mg, 0.0276 mmol, previously azeotroped twice with benzene) and p-lactam 24 (27.5 mg, 0.0721 mmol, previously azeotroped twice with benzene) in THF (1.4 mL), prepared from the Ojima- Holton protocol (Holton, R.A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M. ; Park, Y.H.; 'Sun, C. M. ; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), at 0 C was added NaN(SiMe3)2 (0.066 mL of a 1.0 M solution in THF, 0.066 mmol) dropwise. The resulting solution was stirred for 0.5 h and poured into a mixture of CH2Cl2 (10 mL) and aqueous NH4Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2C12 (2 x 5 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 20 # 30 % ethylacetate in hexanes) to give 34 (1.1 mg, 6%) and 35 ( 17.3 mg, 61% based on 94% conversion) as a white solid. Physical Data for DiTES taxoid 35. Rf= 0.86 (silica, 50% ethylacetate in hexanes) ; IR (film) #max 3519, 3437, 2953, 2879, 1726, 1666, 1515, 1483, 1369, 1240, 1100, 979, 825, EMI62.1 746 cm-1; H NMR (500 MHz, CDC13) s 8.3 2 (dd, J= 3 .0, 1.2 Hz, 1H, thiophene), 7.76-7.73 (m, 2H), 7.60 (dd, J = 5.0, 1.2 Hz, 1H, thiophene), 7.52-7.29 (band, 9H). 7.10 (d, J= 9.0 Hz, 1H, NH), 6.44 (s, 1H, 10-H), 6.26 (br t, J= 9.0 Hz, 1H, 13-H), 5.72 (dd, J= 9.0, 2.0 Hz, 1H, 3'-H), 5.61 (d, J= 7.0 Hz, 1H, 2-H), 4.95 (dd, J= <Desc/Clms Page number 63> 9.5, 2.0 Hz, 1H, 5-H), 4.70 (d, J= 2.0 Hz, 1H, 2'-H), 4.48 (dd, J= EMI63.1 10.5, 6.5 Hz, 1H, 7-H), 4..-),7 i(d, j = 8.5 Hz, 1H, 20-H), 4.23 (d, J= 8.5 Hz, 1H, 20-H), 3.81 (d, J= 7.0 Hz, 1H, 3-H), 2.56-2.49 (m, 1H, 6-H),' 2.54 (s, 3H, Me), 2.35 (dd, J= 15.5,9.0 Hz, 1H, 14-H), 2.17 EMI63.2 (s, 3H, Me),±.07 (dd, j 15.5, 9.0 Hz, 1H, 14-H), 2.03 (d, J= 1.0 Hz, 3H, 18-Me), 1.94-1.87 (m, 1H, 6-H),1.69 (s, 3H, Me), 1.68 (s, 1H, OH), 1.20 (s, 3H, Me), 1.18 (s, 3H,'Me), 0.93 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.81 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.63-0.53 (band, 6H, OSi(CH?CH3)3), 0.52-0.36.(band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m/ e 1220.3675, M + Cs+ calcd for C57H77O14SSi2N 1220.3658. Taxoid 3 6. A solution of silyl ether 35 ( 17 .3 mg, 0.0159 mmol) in THF (0.6 mL) at 25 C was treated with HF#pyridine (0.150 mL) and stirred, for 2 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHC03 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washd with brine (5 mL), dried (MgSO), concentrated, and purified by preparative TLC (silica, 25% ethylacetate in petroleum ether) to give 3 6 (7.7 mg, 56%) as a colorless film. Preparation of Taxoid 3 6. Rf= 0.11 (silica, 50% ethylacetate in hexanes); IR (film) #max 3496, 3434. 2940, '1723, 1648, 1519, 1370, 1243, 1071,975 cm-1; 1H NMR (500 MHz, CDC13) 88.32 (dd, J= 3.0, 1.0 Hz, 1H, thiophene), 7.75-7.72 <Desc/Clms Page number 64> (m, 2H), 7.60 (dd, J = 5.0, 1.0 Hz, 1H, thiophene), 7.53-7.33 (band, 9H), 6.95 (d, J= 9.0 Hz, 1H, NH), 6.28-6.23 (m, 2H, 10-H, 13-H), 5.81 (dd, J= 9.0, 2.0 Hz, 1H, 3'-H), 5.58 (d, J= 7.0 Hz, 1H, 2-H), 4.95 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.80 (dd, J= 4.5, 2.0 Hz, 1H, 2'-H), 4.41 (br t, J= 7.5 Hz, 1H, 7-H), 4.36 (d, J= 8.5 Hz, 1H, 20-H), 4.22 (d, J= 8.5 Hz, 1H, 20-H), 3.78 (d, J = 7.0 Hz, 1H, 3H), 3.49 (d, J= 4.5 Hz, 1H, 2'-OH), 2.55 (ddd, J= 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.45 (br s, 1H, OH), 2.40 (s, 3H, Me), 2.34 (dd, J= 15.5, 9.0 Hz, 1H, 14-H), 2.25 (dd, J= 15.5, 9.0 Hz, 1H, 14-H), 2.24 (s, 3H, Me), 1.89 (ddd, J= 14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.81 (d, J= 2.0 Hz, 3H, 18-Md), 1.74 (br s, 1H, OH), 1.67 (s, 3H, Me), 1.24 (s, EMI64.1 3H, Me), 1.13 (s, 3H, Me); FAB HRMS (NBA / Csl) m / e 992.1940, M + Cs+ calcd for C45H49014NS 992.1928. <Desc/Clms Page number 65> Preparation of 2-pyridinyl-C-2 taxol (40) EMI65.1 Acetate 37. A solution of alcohol 15 (23.2 mg, 0.0353 mmol) and 4-dimethylaminopyridine (DMAP, 12.9 mg, 0.106 mmol) in CH2C12 (1.5 mL) at 25 C was treated with acetic anhydride (0.126 mL, 1.34 mmol) and stirred for 2 h. The reaction mixture was diluted with ethylacetate (5 mL), treated with aqueous NaHC03 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4). concentrated, and purified by flash chromatography (silica, 70 # 100% ethylacetate in petroleum ether) to give 37 ( 19.0 mg, <Desc/Clms Page number 66> 77%) as an amorphous solid. Preparation of Acetate 37. Rf = 0.58 (silica, ethylacetate); IR (film) #max 3482, 2954, 2881, 1730,1675, 1370,1304, 1231, 1118, 987, 823, 739 cm-1; 1H NMR (500 MHz, CDC13) 8 8.77 (ddd, J = 4.5, 1.7,1.0 Hz, 1H, pyridine), 8.05 (ddd, J= 8.0, 1.0, 1.0 Hz, 1H, pyridine), 7.89 (ddd, J= 8.0,8.0, 1.7 Hz, 1H, pyridine), 7.53 (ddd, J = 8.0, 4.5,1.0 Hz, 1H, pyridine), 6.59 (s, 1H, 10-H), 5.65 (dd, J= 6.6,1.0 Hz, 1H, 2-H), 4.92 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.48 (dd, J = 10.5, 7.0 Hz, 1H, EMI66.1 7-H), 4.35 (d, J= 8.5 Hz, H, 20-H), 4.26 (dd, J= 8.5, 1.0 Hz, 1H, 20-H), 3.91 (d, J= 6.5 Hz, 1H, 3-H), 3.00 (d, J= 20.0 Hz, 1H, 14H), 2.71 (dd, J= 20.0, 1.0 Hz, 1H, 14-H), 2.54 (ddd, J= 14.5, 9.5, 7.0 Hz, 1H, 6-H), 2.53 (s, 1H, OH), 2.23 (s, 3H, Me), 2.18 (s, 3H, Me), 2.14 (s, 3H, Me), 1.88 (ddd, J = 14.5, 10.5,2.0 Hz, 1H, 6-H), 1.70 (s, '3H, Me), 1.27 (s, 3H, Me), 1.20 (s, 3H, Me), 0.92 (t, 9H, J = 8.0 Hz, OSi(CH2CH3)3), 0.64-0.52 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m/ e 832.2139, M + Cs- calcd for C36H49O11NSi 832.2129. Alcohol 38. A solution of enone 37 (47.6 mg, 0.0680 mmol) in MeOH-THF (5 : 1, 3.8 mL) at 0 C was treated with NaBH4 (46.0 mg, 1.22 mmol, added by portions) and stirred for 1.5 h. The reaction mixture was diluted with ethylacetate (10 mL), treated with aqueous NH4Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), <Desc/Clms Page number 67> concentrated, and purified 'by flash chromatography (basic alumina, ethylacetate - 10% MeOH in ethylacetate) to give 27f (28.0 mg, 59%) as an amorphous solid. Physical Data for Alcohol 38. Rf= 0.3 6 (silica, ethylacetate) ; IR (film) \'max 3487, 2951, 2880, 1736,1583, 1369,1307, 1236, 1132, 983, 824, 739 cm-1; 1H NMR (500 MHz, CDC13) # 8.79 (dm, J= 4.5 Hz, 1H, pyridine), 8.13 (br d, J= 7.5 Hz, 1H, pyridine), 7.88 (ddd, J = 7.5, 7.5,1.7 Hz, 1H, pyridine), 7.51 (ddd, J = 7.5, 4.5, 1.0 Hz, 1H, pyridine), 6.46 (s, 1H, 10-H), 5.64 (d, J= 7.0 Hz, 1H, 2-H), 4.96 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.85 (br t, J= 8.0 Hz, 1H, 13-H), 4.49 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.31 (d, J= 8.0 Hz, 1H, 20-H), 4.25 (d, J= 8.0 Hz, 1H, 20-H), 3.89 (d, J= 7.0 Hz, 1H, 3-H), 2.5 3 (ddd, J= 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.36-2.11 (m, 2H, 14-CH2), 2.25 (s, 3H, Me), 2.19 (d, J= 1.0 Hz, 3H, 18-Me), 2.18 (s, 3H, Me), 1.88 (ddd, J= 14.0,10.5, 2.5 Hz, 1H, 6-H), 1.70 (s, 3H, Me), 1.20 (s, 3H, Me), 1.05 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.65-0.51 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA / CsI) m / e EMI67.1 834.2311, M + CsT calcd for C6H51011NSi 834.2286. DiTES taxoid 39. To a solution of alcohol 38 (10.3 mg, 0.0147 mmol, previously azeotroped twice with benzene) and -lactam 24 ( 17.0 mg, 0.0446 mmol, previously azeotroped twice with benzene) in THF (0.75 mL) at 0 C, prepared from the Ojima-Holton protocol (Holton, R.A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M. ; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; <Desc/Clms Page number 68> Zhao, M. ; Zucco, M. ; Park, Y.H.; Sun, C. M.; Brigaud, T. Tetrahedron' 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.038 mL of a 1.0 Msolution in THF, 0.038 mmol) dropwise. The resulting solution'was stirred for 20 inin and poured into a mixture of ethylacetate ,(10 mL) and aqueous NH4CI (5 mL). The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4). concentrated, and purified by preparative TLC (silica, 60% ethylacetate in petroleum ether) to give 3 9 (2.7 mg, 17%) as a film. Physical Data for DiTES taxoid 39. Rf = 0.28 (silica, 50% ethylacetate in hexane); IR (film) #max 3429, 2952, 2927, 2878,1728,1662,1585,1369,1236,1124,1016,984,742 cm- 1; 1H NMR (500 MHz, CDC13) 8 8.78 (br d, J = 4.5 Hz, 1H, pyridine), 8.21 (d, J= 8.0 Hz, 1H, pyridine), 7.95 (ddd, J= 8.0, 8.0, 1.7 Hz, 1H, pyridine), 7.75-7.70 (m, 2H), 7.54-7.22 (band, 9H), 7.12 (d, J= 9.0 Hz, 1H, NH), 6.45 (s, 1H, 10-H), 6.27 (br t, J = 9.0 Hz, 1H, 13-H), 5.73-5.67 (m, 2H, 2-H, 3'-H), 4.95 (dd, J= 9.5,2.0 Hz, 1H, 5-H), 4.70 (d, J= 2.0 Hz, 1H, 2'-H), 4.48 (dd, J= 10.5, 6.5, 1H, 7-H), 4.32 (br s, 2H, 20-CH?), 3.85 (d, J= 7.0 Hz, 1H, 3-H), 2.56-2.48 (m, 1H, 6-H), 2.52 (s, 3H, Me), 2.40 (dd, J= 15.0 Hz, 9.5 Hz, 1H, 14-H), 2.20-2.12 (m, 2H, 14-H, OH), 2.18 (s, 3H, Me), 2.04 (s, 3H, Me), 1.92 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, EMI68.1 6-H), 1.72 (s, 3H, Me), 1.22 (s, 3H, 1\le), 1.19 (s, 3H, l\1e), 0.93 (r, J = 8.0 Hi, 9H, OSi(CH2CH3)3), 0.81 (t, J = 8.0 Hz, 9H, EMI68.2 OSi(CH2CH3)3), 0.64-0.34 (band, 12H, OSi(CH2CH3)3); FAB HRMS <Desc/Clms Page number 69> EMI69.1 (NBA / CsI) m / e 1215.4065, M + Cs+ calcd for CSSH7S01.+N2Si2 1215.4046. Taxoid 40. A solution of silyl ether 39 ( 2.7 mg, 0.00249 EMI69.2 mmol) in THF ( 0.4 mL) at 2 5 OC was treated with HF pyridine (0.170 mL) and stirred for 3 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHC03 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO), concentrated, and purified by preparative TLC (silica, ethylacetate) to give 40 (0.8 mg, 38%) as a colorless film. Physical Data for Taxoid 40. Rf = 0.54 (silica, ethylacetate); 1H NMR (500 MHz, CDC13) # 8.80 (br d, J = 4.5 Hz, 1H, pyridine), 8.22 (d, J= 7.5 Hz, 1H, pyridine), 7.93 (ddd, J= 7.5, 7.5, 1.5 Hz, 1H, pyridine), 7.75-7.71 (m, 2H), 7.54-7.30 (band, 9H), 6.98 (d, J= 9.0 Hz, 1H, NH), 6.30-6.24 (m, 2H, 10-H, 13-H), 5.82 (dd, J= 9.0, 2.5 Hz, 1H, 3'-H), 5.67 (d, J= 7.0 Hz, 1H, 2-H), 4.95 (dd, J = 10.0, 2.0 Hz, 1H, 5-H), 4.81 (dd, J = 4.5, 2.5 Hz, 1H, 2'-H), 4.41 (ddd, J= 11.0,7.0, 4.5 Hz, 1H, 7-H), 4.31(s, 2H, 20-CH2), 3.81 (d, J= 7.0 Hz, 1H, 3-H), 3.52 (br s, 1H, OH), 3.50 (d, J= 4.5 Hz, 1H, 2'-OH), 2.56 (ddd, J= 14.5, 9.5, 7.0 Hz, 1H, 6-H), 2.46 (d, J= 4.0 Hz, 1H, 7-OH), 2.43-2.30 (m, 2H, 14CH2), 2.38 (s, 3H, OAc), 2.25 (s, 3H, OAc), 1.90 (ddd, J= 14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.81 (s, 3H, Me), 1.71 (s, 3H, Me), 1.26 (s, 3H, Me), 1.15 (s, 3H, Me). <Desc/Clms Page number 70> EMI70.1 Prepartion of 3-pyridinyl;C=2=taxol (44) EMI70.2 Acetate 41. A solution of alcohol 17 (42.9 mg, 0.0652 mmol) and 4-dimethylaminopyridine (DMAP, 23.9 mg, 0.196 mmol) in CH2C12 (2.8 mL) at 25 C was treated with acetic anhydride (0.235 mL, 2.49' mmol) and stirred for 2 h. The reaction mixture was diluted with ethylacetate (5 mL), treated with aqueous NaHC03 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, <Desc/Clms Page number 71> ethylacetate) to give 41 (43.5 mg, 95%) as a white solid. Physical Data for Acetate, 41. Rf = 0.61 (silica, ethylacetate) ; IR (film) #max 3470,3327, 2955, 2881, 1731, 1675, 1592, 1370,1279, 1229, 1108, 822,738 cm-1; 1H NMR (500 MHz, CDC13) # 9.23 (br s, 1H, pyridine), 8.79 (br s, 1H, pyridine), 8.30 (ddd, J= 8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.43 (dd, J = 8.0, 5.0 Hz, 1H, pyridine), 6.58 (s, 1H, 10-H), 5.70 (dd, J= 6.5, 1.0 Hz, 1H, 2-H), 4.91 (dd, J= 9.5,2.0 Hz, 1H, 5-H), 4.47 (dd, J= 10.5, 7.0 Hz, 1H, 7-H), 4.28 (d, J= 8.0 Hz, 1H, 20-H), 4.11 (d, J = 8.0 Hz, 1H, 20-H), 3.91 (d, J = 6:5 Hz, 1H, 3-H), 2.93 (d, J = 20.0 Hz, 1H, 14-H), 2.68: ,(dd, J= 20.0, 1.0 Hz, 1H, 14-H), 2.53 (ddd, J= 14.5,9.5, 7.0 Hz, 1H, 6-H), 2.24 (br s, 1H, OH), 2.22 (s, 3H, Me), 2.18 (s, 3H, Me), 2.17 (s, 3H, Me), 1.85 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-fI), 1.66 (s, 3H, Me), 1.26 (s, 3H, Me), 1.18 (s, 3H, Me), 0.90 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.63-0.51 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m / e 832.2145, M + Cs+ calcd for C36H49O11NSi 832.2129. Alcohol 42. A solution of enone 41 (39.8 mg, 0.0569 mmol) in MeOH-THF (5 : 1, 3.1 mL) at 0 C was treated with NaBH4 (65.0 mg, 1.72 mmol, added by portions) and stirred for 1.5 h. The reaction mixture was diluted with ethylacetate (10 mL), treated with aqueous NH4Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, <Desc/Clms Page number 72> ethylacetate) to give 41 (3.7 mg,'9%) and 42 (24.3 mg, 67% based on 91% conversion) as an amorphous, solid. Physical Data for Alcohol 42. Rf = 0.42 (silica, ethylacetate) ; IR (film) #max 3490, '2953, 2881, 1727, 1592, 1369,1235, 1110, 822, 740 cm-1; 1H NMR (50'0 MHz, CDC13) # 9.30 (d, J= 2.0 Hz, 1H, pyridine), 8.81 (dd, J= 5.0, 2.0 Hz, 1H, pyridine), 8.35 (ddd, J= 8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.44 (dd, J= 8.0, 5.0 Hz, 1H, pyridine), 6.46 (s, 1H, 10-H), 5.64 (d, J= 7.0 Hz, 1H, 2-H), 4.96 (dd, J = 9.5, 1.5 Hz, 1H, 5-H), 4.83 (br dd, J= 12.5, 7.5 Hz, 1H, 13-H), 4.49 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.28 (d, J= 8.0 Hz, 1H, 20-H), 4.15 (d, J= 8.0 Hz, 1H, 20-H), 3.89 (d, J = 7.0 Hz, 1H, 3-H), 2.53 (ddd, J = 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.30-2.20 (m, 2H, 14-CH2), 2.28 (s, 3H, Me), 2.19 (d, J= 1.0 Hz, 3H, 18-Me), 2.18 (s, 3H, Me), 1.,87 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.68 (s, 3H, Me), 1.63 (br s, 2H, OH, OH), 1.19 (s, 3H, Me), 1.04 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH?CH3)3), 0.64-0.51 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m e 834.2270, M + Cs+ calcd for C36H51O11NSi 834.2286. DiTES taxoid 43 . To a solution of alcohol 42 (12.6 mg, 0.018 mmol, previously azeotroped twice with benzene) and lactam 24 ( 17.0 mg, 0.0446 mmol, previously azeotroped twice with benzene) in THF (0.97 mL) at 0 C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, EMI72.1 164568q; Ojima, L; Habus, L; Zhao, 1\1.; Georg, G. I.; Jayasmghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M. ; Zucco, ML; Park, Y.H.; Sun, C. ML; Brigaud, T. <Desc/Clms Page number 73> Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3) 2 (0.054 mL of a 1.0 M solution in THF, 0.054 mmol) dropwise. The resulting solution was stirred for 0.5 h and poured into a mixture of ethylacetate (10 mL) and aqueous NH4Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 50 - 95% ethylacetate in hexanes) to ,give 42 (1.0 mg, 8%) and 4 3 (8.6 mg, 48% based on, 92% conversion) as a white solid. Physical Data for DiTES taxoid 43. Rf = 0.40 (silica, 50% ethylacetate in hexanes); IR (film) #max 3433, 2955, 2880, 1730,1662,1370,1238,1112,1018,985,824,740 cm-1; 1H NMR (500 MHz, CDC13) # 9.34 (d, J = 2.0 Hz, 1H, pyridine), 8.82 (dd, J = 5 .0, 2.0 Hz, 1H, pyridine), 8.4 2 (ddd, J= 8.0, 2 .0, 2 .0 Hz, 1H, pyridine), 7.74-7.69 (m, 2H), 7.51-7.20 (band, 9H), 7.08 (d, J= 9.0 Hz, 1H, NH), 6.46 (s, 1H, 10-H), 6.22 (br t, J= 9.0 Hz, 1H, 13-H), 5.74-5.66 (m, 2H, 2-H, 3'-H), 4.95 (dd, J= 9.5,2.0 Hz, 1H, 5-H), 4.70 (d, J = 2.0 Hz, 1H, 2'-H), 4.48 (dd, J = 10.5,6.5 Hz, 1H, 7-H),4.30 (d, J= 8.0 Hz, 1H, 20-H), 4.21 (d, J= 8.0 Hz, 1H, 20H), 3.86 (d, J= 7.0 Hz, 1H, 3-H), 2.58-2: 48 (m, 1H, 6-H), 2.54 (s, 3H, Me), 2.40 (dd, J= 15.5, 9.0 Hz, 1H, 14-H), 2.17 (s, 3H, Me), 2.14 (dd, J = 15.5, 9.0 Hz, 1H, 14-H),2.03 (br s, 3H, Me), 1.95- 1.86 (m, 1H, 6-H), 1.73 (s, 4H, Me, OH), 1.22 (s, 3H, Me), 1.18 (s, EMI73.1 3H, Me), 0.93 (t, J= 8.0 Hz, 9H, OSi(CHCH;);), 0.8 2 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.65-0.37 (band, 12H, OSi(CH2CH3)3, OSi(CH?CH3)3); FAB HRMS (NBA/ CsI)'m / e 1215.4066, M + <Desc/Clms Page number 74> EMI74.1 Cs+ calcd for C5 8H780 j 4N2S12 1215.4046. Taxoid 44. A solution of silyl ether 43 (6.4 mg, 0.0059 EMI74.2 mmol) in THF (0.4 mL) at 25 oc was treated with HF. pyridine (0.160 mL) and stirred for 1.25 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHC03 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, ethylacetate) to give 44 (3.8 mg, 75%) as a colorless film. Physical Data for Taxoid 44. Rf= 0.59 (silica, ethylacetate) ; IR (film) #max 3396, 2928, 1728, 1644, 1371, EMI74.3 1273, 1241, 1111, 1071 cm-1; I'H.NNM (500 MHz, CDC13) 8 9.34 (br s, 1H, pyridine), 8.83 (br d, J = 3.5 Hz, 1H, pyridine), 8.41 (br d, J = 8.0 Hz, 1H, pyridine), 7.75-7.68 (m, 2H), 7.53-7.34 (band, 9H), 6.91 (d, J= 9.0 Hz, 1H, NH), 6.27 (s, 1H, 10-H), 6.23 (br t, J= 9.0 Hz, 1H, 13-H), 5.78 (dd, J= 9.0, 2.5 Hz, 1H, 3'-H), 5.69 (d, J= 7.0 Hz, 1H, 2-H), 4.95 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.79 (dd, J= 5.5, 2.5 Hz, 1H, 2'-H), 4.41 (ddd, J= 11.0, 6.5, 4.0 Hz, 1H, 7-H), 4.29 (d, J= 8.5 Hz, 1H, 20-H), 4.20 (d, J= 8.5 Hz, 1H, 20-H), 3.82 (d, J= 7.0 Hz, 1H, 3-H), 3.54 (d, J= 5.5 Hz, 1H, 2'-OH), 2.56 (ddd, J= 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.49 (d, J= 4.0 Hz, 1H, 7-OH), 2.43-2.26 (m, 2H, 14-CH2), 2.3S (s, 3H, Me), 2.24 (s, 3H, Me), 1.89 (ddd, J = 14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.83 (s, 1H, OH), 1.82 (s, 3H, Me), 1.69 (s, 3H, Me), 1.25 (s, 3H, Me), 1.14 <Desc/Clms Page number 75> (s, 3H, Me) ; FAB HRMS (NBA / CsI) m/ e 987.2325, M + Cs+ calcd for C46H50O14N2 987.2316. EMI75.1 Preparation of 4-N,,N-dimethylaniline-C-2 taxol (48) EMI75.2 Acetate 45. A solution of. alcohol 18 (50.0 mg, 0.0714 mmol) and 4-dimethylaminopyridine (DMAP, 26.0 mg, 0.213 mmol) in CH2C12 (3.0 mL) at 25 C was treated with acetic anhydride (0.250 mL, 2.65 mmol) and stirred for 2.5 h. The reaction mixture was diluted with CH2C12 (10 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 25 mm. The organic layer was separated and the aqueous layer was extracted with CH2C12 (2 x 10 mL). The combined organic layer <Desc/Clms Page number 76> was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10% ethylacetate in benzene) to give 45 (41.0 mg, 77%) as an amorphous solid. Physical Data for Acetate 45. Rf = 0.27 (silica, 35% ethylacetate in hexanes) ; IR (film) vmax 3425, 2945, 1722, 1674, 1605, 1365, 1275,1232, 1179, 1094; 1H NMR (500 MHz, CDC13) # 7.89 (d, J= 9.0 Hz, 2H, Ar), 6.64 (d, J= 9.0 Hz, 2H, Ar), 6.56 (s, 1H, 10-H), 5.64 (d, J= 6.5 Hz, 1H, 2-H), 4.90 (br d, J = 8.0 Hz, 1H, 5-H), 4.45 (dd, J= 10.5, 7.0 Hz, 1H, 7-H), 4.36 (d, J= 9.0 Hz, 1H, 20-H), 4.11 (d, J= 9.0 Hz, 1H, 20-H), 3.85 (d, J= 6.5 Hz, 1H, 3-H), 3.05 (s, 6H, NMe?), 2.90 (d, J= 20.0 Hz, 1H, 14-H), 2.62 (d, J= 20.0 Hz, 1H, 14-H), 2.51 (ddd, J= 14.0, 8.0, 7.0, 1H, 6-H), 2.20 (s, 3H, Me), 2.16 (s, 3H, Me), 2.15 (s, 3H, Me), 2.04 (s, 1H, OH), 1.84 (ddd, J= 14.0,10.5, 2.0 Hz, 1H, 6-H), 1.63 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), 0.89 (t, J= 8.0 Hz, 9H, OSi(CH?CH3)3), 0.58-0.53 (band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m / e 874.8589, M + Cs+ calcd for C39H55O11NSi 874.8594. Alcohol 46. A solution of enone 45 (40.0 mg, 0.0539 mmol) in MeOH-THF (5.8 : 1, 4.1 mL) at 0 C was treated with NaBH4 (30.2 mg, 0.80 mmol, added by portions), stirred for 1 h, allowed to warm to 25 C and stirred for 1.5 h. The reaction mixture was diluted with CH2Cl2 (15 mL), treated with aqueous NH4Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine <Desc/Clms Page number 77> (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 25 # 50% ethylacetate in petroleum ether) to give 45 (6.0 mg, 15%) and 46 (30.0 mg, 88% based on 85% conversion) as an amorphous solid. Physical Data for Alcohol 46. Rf = 0.30 (silica, 50% ethylacetate in petroleum ether); 1H NMR (500 MHz, CDC13) # 7.93 (d, J= 9.0 Hz, 2H, Ar), 6.64 (d, J= 9.0 Hz, 2H, Ar), 6.42 (s, 1H, 10-H), 5.57 (d, J= 7.0 Hz, 1H, 2-H), 4.94 (br d, J= 8.0 Hz, 1H, 5-H), 4.83-4.75 (m, 1H, 13-H), 4.46 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.34 (d, J= 8.5 Hz, 1H, 20-H), 4.13 (d, J= 8.5 Hz, 1H, 20H), 3.82 (d, J= 7.0 Hz, 1H, 3-H), 3.04 (s, 6H, Me2N), 2.54-2.44 (m, 1H, 6-H), 2.26 (s, 3H, Me), 2.23 (d, J= 7.5 Hz, 2H, 14-CH2), 2.16 (s, 6H, Me, Me), 2.08 (d, J= 4.5 Hz, 1H, OH), 1.89-1.80 (m, 2H, 6-H, OH), 1.64 (s, 3H, Me), 1.16 (s, 3H, Me), 1.01 (s, 3H, Me), 0.89 (t, J = 8.5 Hz, 9H, OSi(CH?CH3)3), 0.62-0.48 (band, 6H, OSi(CH2CH3) 3). DiTES taxoid 47. To a solution of alcohol 46 ( 14.0 mg, 0.0188 mmol, previously azeotroped twice with benzene) and P-lactam 24 (25.0 mg, 0.0656 mmol, previously azeotroped twice with benzene) in THF (0.75 mL) at 0 C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M. ; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991,56, 1681-1683; Ojima, I.; Habus, I.; EMI77.1 Zhao, M.; Zucco, W.; Park, '.H.; Sun, C. 1.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.056 mL of a 1.0 M solution in THF, 0.056 mmol) dropwise. <Desc/Clms Page number 78> The resulting solution was stirred for 20, min and poured into a mixture of CH2cL2 (10 mL) and aqueous NH4Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2C12 (2 x 5 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 10 # 15% ethylacetate in benzene, then 50% ethylacetate in petroleum ether) to give 47 (12.0 mg, 57%) as a white solid. Physical Data for,DiTES taxoid 47. Rf = 0.26 (silica, 15% ethylacetate in PhH); IR (film) vmax 3425,2946, 2882, 1722, 1669,1600, 1365, 1275, 1238, 1179, 1094 cm-1; IH NMR (500 MHz, CDC13) # 7.96 (d, 'j = 9.0 Hz, 2H, Ar), 7.77-7.72 (m, 2H), 7.54-7.26 (band, 8H), 7.12 (d, J= 8.5 Hz, 1H, NH), 6.69 (d, J = 9.0 Hz, 2H), 6.43 (s, 1H, 10-H), 6.23 (br t, J= 9.0 Hz, 1H, 13-H), 5.68-5.63 (m, 2H, 2-H, 3'-H), 4.93 (br d, J = 8.0 Hz, 1H, 5-H), 4.67 (d, J= 2.0 Hz, 1H, 2'-H), 4.45 (dd, J= 10.5 Hz, 6.5 Hz, 1H, 7H), 4.36 (d, J= 8.5 Hz, 1H, 20-H), 4.20 (d, J= 8.5 Hz, 1H, 20-H), 3.78 (d, J = 7.0 Hz, 1H, 3-H), 3.04 (s, 6H, Me2N), 2.55-2.46 (m, 1H, 6-H), 2.53 (s, 3H, OAc), 2.36 (dd, J = 15.5, 9.0 Hz, 1H, 14-H), 2.15 (s, 3H, Me), 2.09 (dd, J= 15.5,'9.0 Hz, 1H, 14-H), 2.00 (d, J = 1.0 Hz, 3H, Me), 1.92-1.84 (m, 2H, 6-H, OH), 1.67 (s, 3H, Me), 1.20 (s, 3H, Me), 1.16 (s, 3H, Me), 0.90 (t, J = 8.0 Hz, 9H, OSi(CH?CH3)3), 0.79 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.63-0.35 EMI78.1 (band, 12 H, 'OSi(CH?CH3 ) ; ); FAB HRMS (NBA Cs1) m ; e 1257.4503, M + Cs+ calcd for C61H84O14N2Si2 1257.4515. Taxoid 48. A solution of silyl ether 47 (12.0 mg, 0.0107 <Desc/Clms Page number 79> mmol) in THF ( 1.0 mL) at 25 C was treated with HF#pyridine (0.05 mL) and stirred for 1.5 h. The reaction mixture was poured into a mixture of ethylacetate, (10 mL) and aqueous NaHC03 (5 mL) and the .resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 50 # 75% ethylacetate'in,petroleum .ether) to give 48 (8.0 mg, 84%) as a colorless film. Physical Data for Taxoid 48,. Rf = 0.44 (silica, 75% ethylacetate in petroleum ether); IR (film) #max 3414,2914, EMI79.1 2850, 1722, 1664, 1660, 1371, 1275,1243, 1179 cm-1; IH NMR EMI79.2 (500 MHz, CDC13) 5 7.95 (d, J= 9.0 Hz,'2H), 7.77-7.72 (m, 2H), 7.55-7.30 (band, 8H), 7.03 (d J = 9.0 Hz, 1H, NH), 6.67 (d, J = 9.0 Hz, 2H), 6.24 (s, 1H, 10-H), 6.20 (br t, J= 9.0 Hz, 1H, 13-H), 5.76 (dd, J= 9.0, 2.5 Hz, 1H, 3'-H), 5.62 (d, J= 7.0 Hz, 1H, 2-H), 4.93 (br d, J= 7.5 Hz, 1H, 5-H), 4.76 (dd, J= 5.0,2.5 Hz, 1H, 2'H), 4.37 (ddd, J= 11.5, 6.5, 4.0 Hz, 1H, 7-H), 4.34 (d, J= 8.5 Hz, EMI79.3 1H, 20-H), 4.18 (d, J-- 8.5 Hz, 1H, 20-H), 3.73 (d, J= 7.0 Hz, 1H, 3-H), 3.57 (d, J= 5.0 Hz, 'lH, 2'-OH), 3.04 (s, 6H, Me2N), 2.58- 2.48 (m, 1H, 6-H), 2.44 (d, J = 4.0 Hz, 1H, 7-OH), 2.37 (s, 3H, Me), 2.30-2.25 (m, 2H, 14-CH2), 2.22 (s, 3H, Me), 1.95 (s, 1H, OH), 1.88-1.81 (m, 1H, 6-H), 1.74 (d, J = 1.0 Hz, 3H, Me), 1.65 (s, 3H, Me), 1.21 (s, 3H, Me), 1.11 (s, 3H, Me); FAB HRMS (NBA EMI79.4 Csl) m / e 1029.2760, M + Cs.1- calcd for C49H56N20i4 1029.2786. <Desc/Clms Page number 80> Preparation of 1-naphthalene-C-2-taxol (52) EMI80.1 Acetate 49. A solution of previous alcohol 19 and 4dimethylaminopyridine (DMAP, 100 mg, 0.819 mmol) in CH2C12 (3 mL) at 25 C was treated with acetic anhydride (0.50 mL, 5.30 mmol) and stirred for 3 h. The reaction mixture was diluted with CH2C12 (5 mL), treated with aqueous NaHC03 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2C12 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by preparative TLC (silica, 10% ethylacetate in benzene) to give <Desc/Clms Page number 81> 49 (54.1 mg, 89% from carbonate 7) as.an amorphous solid. Physical Data for ,Acetate 49. Rf = 0.27 (20% ethylacetate in petroleum, ether); IR (film) #max 3416, 2953, 2879, 1726, 1676,1370, 1224, 1089 'cm-; 1H NMR (500 MHz; CDC13) # 8.66 (s, 1H, naphthalene), 8.06 (dd, 1H, J== 9.0,2.0 Hz, naphthalene), 7.98-7.89 (m, 3H; naphthalene), 7.68-7.55 (m, 2H, naphthalene), 6.61 (s, 1H, 10-H), 5.75 (d, J= 7.0 Hz, 1H, 2H), 4.95 (br d, J= 8.0 Hz, 1H, 5-H), 4.50 (dd, J= 10.5, 7.0 Hz, 1H, 7-H), 4.35 (d, J= 8.5 Hz, 1H, 20-H), 4.16 (d, J= 8.5 Hz, 1H, 20H), 3.96 (d, J = 8.5 Hz, 1H, 20-H), 3.96 (d, J = 7.0 Hz, 1H, 3-H), 3.03 (d, J= 20.0 Hz, 1H, 14-H), 2.70 (d, J= 20.0 Hz, 1H, 14-H), 2.61-2.50 (m, 2H, 6-H, OH), 2.27 (s, 3H, Me), 2.24 (s, 3H, Me), 2.21 (s, 3H, Me), 1.91-1.83 (m, 1H, 6-H), 1.70 (s, 3H, Me), 1.30 (s, 3H, Me), 1.20 (s, 3H, Me), 0.93 (t, J = 8.0 Hz, 9H, OSi(CH2CH3)3), 0.66-0.57.(band, 6H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m / e 881.2326, M + Cs+ calcd for C41H52O11Si 881.2333. Alcohol 50. A solution ot enone 49 (54.1 mg, 0.0722 mmol) in MeOH (10 mL) at 25 C was treated with NaBH4 (54.5 mg, 1.44 mmol, added by portions) and stirred for 2.0 h. The reaction mixture was diluted with CH2Cl2 (10 mL), treated with aqueous NH4CI (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 20% ethylacetate in petroleum <Desc/Clms Page number 82> ether) to give 50 (26 mg, 48% ) as an amorphous solid. Physical Data for Alcohol 50. Rf = 0.12 (20% ethylacetate in petroleum ether); IR (film) vmax 3524, 2953, 1719, 1369,' 1231, 1093, 829'cm-1 ; 1H NMR (500 MHz, CDC13) # 8.70 (s, 1H, naphthalene), 8.11 (dd, J = 8.5, 1.5 Hz, 1H, naphthalene), 7.96-7.86 (m, 3H, naphthalene), 7.65-7.54 (m, 2H, naphthalene), 6.45 (s, 1H, 10-H), 5.68 (d, J= 7.0 Hz, 1H, 2- H), 4.98 (br d, J = 8.0 Hz, 1H, 5-H), 4.88-4.81 (m, 1H, 13-H), 4.51 (dd, J= 10.5, 7.0 Hz, 1H, 7-H), 4.34 (d, J = 8.5 Hz, 1H, 20- H), 4.19 (d, J= 8.5 Hz, 1H, 20-H), 3.93 (d, J= 7.0 Hz, 1H, 3-H), 2.58-2.50 (m, 1H, 6-H), 2.41-2.14 (m, 3H, 14-CH?, 13-OH), 2.37 (s, 3H, Me), 2.21 (br s, 3H, Me), 2.19 (s, 3H, Me), 1.92-1.84 (m, 1H, 6-H), 1.72 (s, 1H, OH) 1.71 (s, 3H, Me), 1.22 (s, 3H, Me), 1.05 (s, 3H, Me), 0.93 (t,'J= 8.0 Hz, 9H, OSi(CH?CH3)3), 0.65-0.51 EMI82.1 (band, 6H, OSi(CH?¯CH3)3); FAB HRMS (NBA / CsI) m / e 883.2484, M + Cs+ calcd for C41H54O11Si 883.2490. DiTES taxoid 51. To a solution of alcohol 50 (20.0 mg, 0.0266 mmol, previously azeotroped twice with benzene) and -lactam 24 (20.0 mg, 0.0525 mmol, previously azeotroped twice with benzene) in THF (1.1 mL) at -78 C, prepared from the Ojima-Holton protocol (Holton, R.A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1'683; Ojima, I.; Habus, I.; Zhao, M. ; Zucco, M.; Park, Y.H.; Sun, C. M.; Brigaud, T. EMI82.2 Tetrahedron 1992, 48, 6985-7012), was added NaN(Sil\4e-'))2 (0.065 mL of a 1.0 M solution in THF, 0.065mmol) dropwise. <Desc/Clms Page number 83> The resulting solution was stirred for 10 min and poured into a EMI83.1 mixture of C2C12 ( 10 . mL) and aqueous NH4CI ( 5 mL) . The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 5 mL). The combined organic layer EMI83.2 was washed with brine (5 mL). dried, (MgS04), concentrated, and purified by preparative 'TLC (silica, 20% ethylacetate in petroleum ether) to give 51 (18.7 mg, 62%) as a white solid. Taxoid 52. A solution of silyl ether 51 (18.7 mg, 0.0165 mmol) in THF (2 mL) at 25 C was treated with HF#pyridine (1 mL) and stirred for 1 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHC03 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate ( 2 x 10 mL). The. combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by' preparative TLC (silica, 50% ethylacetate in petroleum ether) to give 52 ( 12.8 mg, 86%) as a colorless film. Physical Data for Taxoid 52. Rf = 0.16 (silica, 50% ethylacetate in petroleum ether); IR (film) #max 3420, 2967, 2896, 1721, 1652, 1519, 1370, 1233, 1073,776 cm-1; 1H NMR (500 MHz, CDC13) 8 8.67 (s, 1H, naphthalene), 8.04 (dd, J= 8.5, 1.5 Hz, 1H, naphthalene), 7.95 (br d, 8.5 Hz, 1H, naphthalene), 7.87 (br d, 9.0 Hz; 1H), 7 .81 ( br d, 8.5 Hz, 1H), 7.65-7.61 (m, 2H), 7.56-7.51 (m, 1H), 7.49-7.22 (band, 9H), 6.94, (d, J= 9.0 Hz, 1H, NH), 6.23-6.16 (m, 2H, 10-H, 13-H), 5.78 (dd, J = 9.0,2.0 Hz, 1H,3'-H), 5.64 (br d, J = 7.0 Hz, 1H, 2-H), <Desc/Clms Page number 84> 4.87 (br d, J= 8.0 Hz, 1H, 5-H), 4.78-4.72 (m, 1H, 2'-H),4.38- 4.31 (m, 1H, 7-H), 4.24 (d, J= 8.5 Hz, 1H, 20-H), 4.16 (d, J= 8.5 Hz, 1H, 20-H), 3.76 (d, J = 7.0 Hz, 1H, 3-H), 3.53 (br s, 1H, OH), 2.52-2.43 (m, 1H, 6-H), 2.42 (d, J= 4.0 Hz, 1H, OH), 2.40 (s, 3H, Me), 2.36 (dd, J= 15.5,9.0 Hz, 1H, 14-H), 2.25 (dd, J= 15.5,9.0 Hz, 1H, 14-H), 2.17 (s, 3H, Me), 1.85-1.77 (m, 2H, 6-H, OH), 1.74 (br s, 3H, Me), 1.63 (s, 3H, Me), 1.17 (s, 3H, Me), 1.09 (s, 3H, Me) ; FAB HRMS (NBA/ CsI) m/ e 1036.2505, M + Cs+ calcd for C51H53NO14 1036.2520 EMI84.1 Preparation of th aether-C-,2 taxol (56) EMI84.2 Acetate 53. A solution of alcohol 23 (25.2 mg, 0.0351 mmol) and 4-dimethylaminopyridine (DMAP, 12.2 mg, 0.0999 <Desc/Clms Page number 85> EMI85.1 mmol) in CH?C1 ( 1.5 mL), at 25 C was treated with acetic anhydride (0.120 mL, 1.27 mmol) and stirred for 1.5 h. The reaction mixture was diluted with CH2Cl2 (5 mL), treated with aqueous NaHC03 (7 mL),.and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in petroleum ether) to give 53 (25.3 mg, 95%) as a colorless oil. Physical Data for Acetate 53. Rf = 0.41 (silica, 10% ethylacetate in benzene, 2. elutions); IR (film) #max 3471, 2954, 2881, 1729, 1675, 1370, 1226, 986, 824, 738 cm-1; 1H NMR EMI85.2 (500MHz,CDCl3) 5 7.38-7.25 (band, 5H, SPh), 6.54 (s, 1H, 10- H), 5.49 (br d, J = 6.5 Hz, 1H, 2-H), 4.90 (dd, J= 9.5,2.0 Hz, 1H, 5-H), 4.42 (dd, J= 10.5, 6.5 Hz,, 1H, 7-H), 4.37 (d, J= 8.0 Hz, 1H, 20-H), 4.17 (d, J= 8.0 Hz, 1H, 20-H), 3.78 (d, J= 6.5 Hz, 1H, 3H), 3.23-3.13 (m, 2H, CH2SPh), 2.78 (d, J = 20.0 Hz, 1H, 14-H), 2.72-2.58 (m, 3H, CH?CH?SPh, 14-H), 2.52 (ddd, J = 14.5, 9.5, 6.5, 1H, 6-H), 2.45 (s, 1H, OH), 2.21 (s, 3H, Me), 2.15 (s, 3H, Me), 2.04 (s, 3H, Me), 1.86 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.62 (s, 3H, Me), 1.23 (s, 3H, Me), 1.19 (s, 3H, Me), 0.91 (t, J= 8.0 Hz, EMI85.3 9H, OSi(CHCH3)3) , 0.64-0.52 (band, 6H, OSi(CHCH3)3); FAB HRMS (NBA/ CsI) m / e 891.2225, M + Cs+ calcd for C39H54O11SSi 891.2210 Alcohol 54. A solution of enone 53 (24.4 mg, 0.032 mmol) in MeOH-THF (5 : 1, 1.9 mL) at 0 C was treated with <Desc/Clms Page number 86> NaBH4 (18.1 mg, 0.48 mmol, added by portions) and stirred for 1.25 h. The reaction mixture was diluted with CH2Cl2 (5 mL), treated with aqueous NH4Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was EMI86.1 extracted with CH2C12 (2 x 5,, p9L) . The qombined organic layer was washed with brine (5 mL), dried (MgS04), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in hexanes) to give 54 (14.6 mg, 60%) as an amorphous solid. Physical Data for Alcohol 54. Rf = 0.11 (silica, 30% ethylacetate in hexanes); IR (film) vmax 3487, 2938,2880, 1729,1586, 1369, 1234, 977, 738 cm- 1; 1H NMR (500 MHz, CDC13) 8 7.40-7.23 (band, 5H, SPh), 6.42 (s, 1H, 10-H), 5.43 (d, J = 7.0 Hz, 1H, 2-H), 4.94 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.85-4.78 (m, 1H, 13-H), 4.43 (dd, J = 10.5, 6.5'Hz, 1H, 7-H), 4.37 (d, J= 8.0 Hz, 1H, 20-H), 4.18 (d, J= 8.0 Hz, 1H, 20-H), 3.74 (d, J= 7.0 Hz, 1H, 3-H), 3.25-3.15 (m, 2H, CH?SPh), 2.71-2.57 (m, 2H, CH2CH2SPh), 2.51 (ddd, J= 14.5, 9.5,6.5 Hz, 1H, 6-H), 2.25 (dd, J = 15.5, 9.5 Hz, 1H, 14-H), 2.16 (s, 3H, Me), 2.15 (d, J= 1.0 Hz, 3H, 18-Me), 2.15 (s, 3H, Me); 2.09 (dd, J= 15.5,7.0 Hz, 1H, 14H), 2.05 (br's, 1H, OH),'1.99-1.96 (m, 1H, OH), 1.86 (ddd, J = 14.5, 10.5,2.0 Hz, 1H, 6-H), 1.63 (s 3H, Me), 1.15 (s, 3H, Me), EMI86.2 1.04 (s, 3H, Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH?CH3);), 0.64-0.50 (band, 6H, Si(CH2CH3)3); FAB HRMS (NBA CsI) m / e 893.2350, EMI86.3 M + Cs+ calcd for C39H011SSi 893.236 i . DiTES taxoid 55. To a solution of alcohol 54 ( 21.8 mg, <Desc/Clms Page number 87> 0.0286 mmol, previously azeotroped twice with benzene) and -lactam 24 (33.0 mg, 0.0866 mmol, previously azeotroped twice with benzene) in THF (1.1 mL) at! 0 C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M. ; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M. ; Zucco, M. ; Park, Y.H.; Sun, C. M. ; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.086 mL of a 1.0 M solution in THF, 0.086 mmol) dropwise. The resulting solution was stirred for 20 min and poured into a mixture of CH2C12 (10 mL) and aqueous NH4CI (5 mL) . The organic layer was separated and the aqueous layer was extracted with CH2C12 (2 x 5 mL). The combined organic layer was washed with brine (5 mL). dried (MgS04), concentrated, and purified by flash chromatography (silica, 15 # 3 0 # 50% ethylacetate in petroleum ether) to give 55 (13.8 mg, 42%) as an amorphous solid. * Physical Data for DiTES taxoid 55. Rf = 0.40 (silica, 30% ethylacetate in hexanes); IR (film) #max 3437, 2952, 2879, 1735, 1662, 1482, 1369, 1236, 1128, 981, 740 cm-1; 1H NMR (500 MHz; CDC13) 8 7.82-7.76 (m, 2H), 7.54-7.16 (band, 13H), 7.11 (d, J= 9.0 Hz, 1H, NH), 6.41 (s, 1H, 10-H), 6.18 ( br t, J= 9.0 Hz, 1H, 13-H), 5.62 (dd, J= 9.0, 2.0 Hz, 1H, 3'-H), 5.49 (d, J= 7.0 Hz, 1H, 2-H), 4.93 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.64 (d, J= 2.0 Hz, 1H, 2'-H), 4.42 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.40 (d, J= 8.0 Hz, 1H, 20-H), 4.2 1 (d, J= 8.0 Hz, 1H, 20-H), 3.70 (d, J = 7.0 Hz, 1H, 3-H), 3.23-3.17 (m, 2H, CH2SPh), 2.78-2.69 (m, 1H, <Desc/Clms Page number 88> HCHCH2SPh), 2.67-2.'57 (m, 1H, HCHCH2SPh), 2.55-2.46 (m, 2H, 6-H, OH), 2.38 (s, 3H, Me), 2.27-2.10' (m, 2H, 14-CH2), 2.16 (s, 3H, Me), 1.98 (d, J= 1.0 Hz, 3H, Me),,1.89 (ddd, J= 14.0, 11.0, 2.0 Hz, 1H, 6-H), 1.64 (s, 3H, Me), ,1.18 (s, 3H, Me), 1.17 (s, 3H., Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH2CH3)3), 0.81 (t, J= 8.0 Hz, 9H, OSi(CH?CH3)3), 0.64-0.36 (band, 12 H, OSi(CH2CH3)3); FAB HRMS (NBA/ CsI) m / e M + Cs+ 1274.4125 calcd for C61H83O14SSi2 1274.4127. Taxoid 56. A solution of silyl ether 55 (8.1 mg, 0.0071 mmol) in THF (0.5 mL)-,at 25 C was treated with HF#pyridine (0.150 mL) and stirred for 3.75 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHC03 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2 x 10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 60% ethylacetate in petroleum ether) to give 5 6 (3.2 mg, 49%) as a colorless film. Physical Data for Taxoid 56. Rf= 0.39 (silica, 60% ethylacetate in petroleum ether) ; IR (film) #max 3426, 2928, 1731,1642, 1371, 1238, 1070, 739, 709 cm-1; 1H NMR (500 MHz, CDCl3) 5 7.80-7.75 (m, 2H), 7.55-7.18 (band, 13 H), 6.94 (d, J= 9.0 Hz, 1H, NH), 6.23 (s, 1H, 10-H), 6.19 (br t, J= 9.0 Hz, 1H', 13-H), 5.74 (dd, J= 9.0, 2.5 Hz, 1H, 3'-H), 5.47 (d, J= 7.0 Hz, 1H, 2-H), 4.93 (dd, J= 9.5, 2.0, Hz, 1H, 5-H), 4.74 (dd, J= 5.0, 2.5 <Desc/Clms Page number 89> Hz, 1H, 2'-H), 4.38 (d, J= 8.0 Hz, 1H, 20-H), 4.35 (ddd, J= 11.0, 6.5 Hz, 4.5 Hz, 1H, 7-H), 4.21 (d, J= 8.0 Hz, 1H, 20-H), 3.67 (d, J = 7.0 Hz, 1H, 3-H), 3.51 (d, J = 5.0 Hz, 1H, 2'-OH), 3.28-3.14 (m, 2H, CH2SPh), 2.77-2.68 m, 1H, HCHCH2SPh), 2.67-2'.59 (m, 1H, HCHCH2SPh), 2.54 (ddd, J= 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.44 (d, J = 4.5 Hz, 1H, 7-OH), 2.36 (dd, J = 15.5, 9.0 Hz, 1H, 14-H), 2.26 (br s, 1H, OH), 2.23 (s, 3H, Me), 2.21 (s, 3H, Me), 2.18 (dd, J= 15.5,9.0 Hz, 1H, 14-H), 1.88 (ddd, J= 14.5, 11.0, 2.0 Hz, 1H, 6H), 1.75 (d, J = 1.0 Hz, 3H, Me), 1.63 (s, 3H, Me), 1.24 (s, 3H, Me), 1.10 (s, 3H, Me); FAB HRMS (NBA/ CsI) m / e 1046.2410, EMI89.1 M + Cs+ calcd for C49H01'4ITS.104G.2398. Preparation of MPA taxoid 57 EMI89.2 MPA taxoid 57. A solution of taxoid 36 (4.3 mg, 0.003 mmol) and triethylamine (0.0033 mL, 0.0237 mmol) in CH2Cl2 (0.2 mL) a't 25 C was treated with 2-fluoro-1- methylpyridinium p-toluenesulfonate ( 2.1 mg, 0.0075 mmol) and stirred for 35 min. The clear colorless solution rapidly turned to a clear pale yellow. The course of the reaction was <Desc/Clms Page number 90> monitored through thin layer chromatography (TLC)(E. Merck RP- 18 silica, 65 tetrabydrofuran: 35 water, UV/phosphomolybidic acid) and after thirty minutes of stirring at ambient temperature,,, judged'complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture was directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give 36 (0.8 mg, 19%) and 57 (4.1 mg, 100% based on 81% conversion) as a colorless film. EMI90.1 Physical Data for taxoid.57 1 H NMR (5 00 /IHz, CDC13) 8 10.5 (d, J = 7.5 Hz, 1H), 8.44 (ddd, J = 9.0,7.5, 2.0 Hz, 1H), 8.33-8.29 (m, 2H), 8.15 (dd, J = 3.0, 1.0 Hz, 1H, thiophene), 8.12 (br d, J= 6.0.Hz, 1H), 7.84 (br d, J = 8.5 Hz, 1H), 7.74-7.69 (m, 2H), 7.53 (dd, J= 5.0, 1.0 HZ, 1H, thiophene), 7.48-7.34 (band, 7H), 7.16-7.12 (m, 1H), 6.53-6.43 (m, 1H, 2'-H), 6.21 (s, 1H, 10H), 6.03 (dd, J= 10.5,8.0 Hz, 1H, 3'-H), 5.82 (br t, J= 9.0 Hz, 1H, 13-H), 5.44 (d, J= 7.0 Hz, 1H, 2-H), 4.90 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.33 (dd, J= 11.0, 6.5 Hz, 1H, 7-H), 4.30 (d, J = 8.0 Hz, 1H, 20-H), 4.15 (d, J= 8.0 Hz, 1H, 20-H), 4.08 (s, 3H, N+Me), 3.68 (d, J= 7.0 Hz, 1H, 3-H), 2.58-2.49 (m, 1H, 6-H), 2.52 (s, 3H, OAc), 2.21 (s, 3H, OAc), 2.04 (s, 3H, OAc), 2.02 (br s, 2H, OH, OH), 1.88 (ddd, J = 14.5, 11.5, 2.0 Hz, 1H, 6-H), 1.78 (br s, 3H, 18-Me), 1.64 (s, 3H, Me), 1.61 (dd, J= 16.0, 7.0 Hz, 1H, 14-H), 1.18 (dd, J = 16.0, 9.0 Hz, 1H, 14-H), 1.13 (s, 3 H, Me), 1.08 (s, 3 H, Me). <Desc/Clms Page number 91> Preparation nf MPA taxoid 58 EMI91.1 MPA taxoid 5 8. A solution of taxoid 3 2 (1.0 equiv.) and triethylamine' (4.7 equiv.) in CH2Cl2 (0.025 M) at 25 C is EMI91.2 treated with 2-fluoro-l-methylpyridinium p-toluenesulfonate from Aldrich Chemical company inc. (1.5 equiv. ) and stirred for 35 minutes. The course of the reaction,was monitored through .., thin layer chromatography (TLC)(E. Merck RP- 18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH4OAc, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give 58 as a colorless film. <Desc/Clms Page number 92> Preparation of MPA taxoid 5 9 EMI92.1 MPA taxoid 59. The synthesis of the taxoid-7-MPA 5 9 differs only slightly from the synthesis of taxoid-2'-MPA 58. The C-2 taxoid 3 2 is dissolved in methylene chloride (.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-1-methyl-pyridinium tosylate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 26.12 ) to give 59 as a colorless film. Preparation of MPA taxoid 60 EMI92.2 <Desc/Clms Page number 93> MPA taxoid 60. The synthesis of the taxoid-7-MPA 60 differs only slightly from the synthesis of taxoid-2'-MPA 57. The C-2 taxoid 36 is dissolved in methylene chloride (.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-1-methyl-pyridinium tosylate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 26.12,) to ,give 60 as a colorless film. Preparation of C-2-taxoid-2'-methyl-pyridinium salts EMI93.1 C-2-taxoid-2'-onium salts 62-66. A solution of taxoid (62-66) 1. (1.0 equiv. ) and triethylamine (4.7 equiv.) in CH2Cl2 (0.025 M) at 25 C is treated with 2-fluoro-lmethylpyridinium p-toluenesulfonate from Aldrich Chemical <Desc/Clms Page number 94> company inc. (1.5 equiv. ) .and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC) (E. Merck RP- 18 silica, 65 tetrahydrofuran: 35 .water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL / min, RT = 26.12) to give (62-66) II. , as a colorless film. Preparation of C-2-taxoid-7-methyl-py&num;dinium salts EMI94.1 C-2-taxoid-7-onium salts 67-72. The; synthesis of the taxoid-7-methyl-pyridinium salts (67-72) II, differs only slightly from the synthesis of taxoid-2'-methyl-pyridinium salts (62-66) II. The C-2 taxoid (67-72) I is dissolved in <Desc/Clms Page number 95> methylene chloride (. 006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-l-methylpyridinium tosylate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP- 18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give (67- 72) II as a colorless film. Preparation of C-2-taxoid-bis-2',7-methyl-pyridinium salts EMI95.1 C-2-taxoid-bis-2',7-onium salts 73-80. The synthesis of C-2-taxoid-bis-2',7-methyl-pyridinium salts I (73-80), differs from the synthesis of taxoid-7-methylpyridinium salts (67-72) II only with respect to reaction time. The C-2 taxoid (73-80) I is dissolved in methylene chloride (.006 M) and treated sequentially with triethylamine (40 EMI95.2 equivalents) and 2-fluoro-l-methyl-pyridinium tosylate (10 <Desc/Clms Page number 96> equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 18 hours. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give (73-80) II as a colorless film. Preparation of C-2-taxoid-2'-benzothiazolium salts EMI96.1 C-2-taxoid-2'-benzothiazolium salts 81-88. A solution of taxoid (81-88) I (1.0 equiv.) and triethylamine (4.7 equi,,,.) in CH2C12 (0.025 M) at 25 C is treated with 2-fluoro-1- methylpyridinium p-toluenesulfonate from Aldrich Chemical company inc. (1.5 equiv.) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC) (E. Merck RP- 18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-mol'ybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, <Desc/Clms Page number 97> # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give (81-88) II. as a colorless film. Preparation of C-2-taxoid-7-benzothiazolium salts EMI97.1 C-2-taxoid-7-benzothiazolium salts (89-96). The synthesis of the taxoid-7-benzothiazolium salts (89-96) II, differs only slightly from the synthesis of taxoid-2'benzothiazolium salts (81-88) II. The C-2 taxoid (89-96) I is dissolved in methylene chloride (.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro- 3-ethylbenzothiazolium tetrafluoroborate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give (89-96) II as a colorless film. <Desc/Clms Page number 98> Preparation of C-2-taxoid-2'-benzoxazolium salts EMI98.1 C-2-taxoid-2'-benzoxazolium salts 97-104. A solution of taxoid 97-104 1.(1.0 equiv. ) and triethylamine (4.7 equiv. ) in CH2C12 (0.025 M) at 25 C is treated with 2-fluoro-lmethylpyridinium p-toluenesulfonate from Aldrich Chemical company inc. (1.5 equiv.) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC)(E. Merck RP- 18 silica, 65 tetrahydrofuran: 35 water, EMI98.2 UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A -- B 0.5 h linear, A: 20% MeOH in 20 mM NH4OAc, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give 97-104 II. as a colorless film. <Desc/Clms Page number 99> Preparation of C-2-taxoid-7-benzoxazolium salts EMI99.1 C-2-taxoid-7-benzoxazolium salts (105-112). The synthesis of the taxoid-7-benzoxazolium salts (105-112) II, differs only slightly from the synthesis of taxoid-2'benzoxazolium salts (97-104) II. The C-2 taxoid (105-112) I is dissolved in methylene chloride (.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-chloro-3ethylbenzoxazolium tetrafluoroborate from Aldrich Company (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL / min, RT = 26.12) to give (105-112) II as a colorless film. <Desc/Clms Page number 100> Preparation of C-2-taxoid-2'-pyrimidinium salts EMI100.1 C-2-taxoid-2'-pyrimidinium salts 113-120. A solution of taxoid (113-120) 1. (1.0 equiv. ) and triethylamine (4.7 equiv. ) in CH2C12 (0.025 M) at 25 C is treated with 2chloro-methyl-pyrimidinium fluoride from Aldrich Company (1.5 equiv. ) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC) (E. Merck RP- 18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP- 18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40AC, B: 100% MeOH, 9 mL/ min, RT = 26.12) to give (113- 120) II. as a colorless film. <Desc/Clms Page number 101> Preparation of C-2-taxoid-7-pyrimidinium salts EMI101.1 C-2-taxoid-7-pyrimidinium salts (121-128). The synthesis of the taxoid-7-pyrimidinium salts (121-128) II, differs only slightly from the synthesis of taxoid-2'pyrimidinium salts (113-120) II. The C-2 taxoid (121-128) I is dissolved in methylene chloride (. 006 M) and treated sequentially with triethylamine (40 equivalents) and 2-chloromethyl-pyrimidinium fluoride from Aldrich Company (10 equivalents), and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A # B 0.5 h linear, A: 20% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL/ min, RT = 2 6.12 ) to give (121-128) II as a colorless film.

Claims: What is claimed is: 1. A water soluble self-assembled helical fibrous nanostructure having a diameter of 60-120 A and a helical twist of 5-10 molecules per turn of an onium salt of a taxo-diterpenoid-2-O-aza-arene. 2. A water soluble self-assembled helical fibrous nanostructure as described in claim 1 wherein the solubility of the onium salt of the taxo-diterpenoid- 2-0-aza-arene exceeds 10-3 moles per liter. 3. A water soluble self-assembled helical fibrous nanostructure as described in claim 3 wherein the 2O-aza-arene is labile to displacement by serum protein. 4. A composition comprising: an aqueous solvent and an onium salt of a taxo-diterpenoid-2-O-azaarene dissolved in said aqueous solvent and capable of forming a water soluble self-assembled helical fibrous nanostructure above a critical aggregation concentration. 5. A composition as described in claim 4 wherein: said water soluble self-assembled helical fibrous nanostructures having a diameter of 60-120 A and a helical twist of 5-10 molecules per turn. 6. A water soluble self-assembled micellular nanostructure having a spherical shape with a diameter of 20-120 A and incorporating an onium salt of a taxo-diterpenoid-2-O-aza-arene. <Desc/Clms Page number 103> 7. A water soluble self-assembled micellular nanostructure as described in claim 6 wherein the solubility of the onium salt of the taxo-diterpenoid- 2-0-aza-arene exceeds 10-3 moles per liter. 8. A composition comprising: an aqueous solvent and an onium salt of a taxo-diterpenoid-2-O-aza- arene dissolved in said aqueous solvent and capable of forming a water soluble self-assembled micellular nanostructure above a critical aggregation concentration upon sonication. 9. A composition as described in claim 8 wherein: said water soluble self-assembled micellular nanostructure having a spherical shape with a diameter of 20-120 A. 10. A process for making a self-assembled micellular nanostructure from an aqueous solution including an onium salt of a taxo-diterpenoid-2-O-aza-arene, said process employing a step of sonicating the aqueous solution at a concentration in excess of a critical aggregation cencentration for the onium salt of the taxo-diterpenoid-2-O-aza-arene. <Desc/Clms Page number 104> 11. A self-assembled helical fibrous nanostructure having a diameter of 60-120 A and a helical twist of 5-10 molecules per turn of an onium salt of a taxo- diterpenoid-Cn,2-0-aza-arene having an aqueous solubility in excees of 10-3 moles per liter, said EMI104.1 onium salt of said taxo-diterpenoid-Cn, 2-0-aza-arene represented by the following formula: EMI104.2 wherein: Cn is selected from the group consisting of C7 and C2'; Rx is selected from the group consisting of Ph and tBuO; R10 is selected from the group consisting of OAc and OH; and RY is C-2 substituent selected from the group consisting of benzyl and the following structures: EMI104.3 ; and R2' and R7 are each selected from the group consisting of OH and an onium salt of a 2- O-aza-arene, with the proviso that at least one of R2' and R7 is said onium salt of the 2-0-aza-arene, said onium salt of the 2-0- <Desc/Clms Page number 105> Consisting of onium salt I and onium salt II represented by the following formulas: EMI105.1 onium salt I onium sait II wherein: Z1 and Z2 are each selected from the group consisting of C and N; Z3 is selected from the group consisting of S and 0; R1 is selected from the group consisting of C1- C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; R2 and R6 are each selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; if Z' is C, then R3is selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6 0-alkyl, OH, halogen, and fused aryl; if Z' is N, then R3is absent; R4 and R8 are each selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, CI-C6 0-alkyl, OH, halogen, and fused aryl; and if Z2 is C, then R5 is selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6 0-alkyl, OH, halogen, and fused aryl; if Z2 is N, then R5 is absent; and <Desc/Clms Page number 106> S' is a counter ion. 12. A self-assembled helical fibrous nanostructure as described in claim 11 wherein: S' is selected from the group of counter ions consisting of Cl-, Br', I', BF4-, C104 , ArS03 , and C1-C6 alkylS03-' 13. A self-assembled helical fibrous nanostructure as described in claim 11 wherein: R2' is said onium salt and R7 is OH. 14. A self-assembled helical fibrous nanostructure as described in claim 13 wherein: Rx i s Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R' is CI-C6 alkyl; R2, R3, R4, and R4 are each H; and S' is selected from the group of counter ions consisting of Cl-, Br', I', BF4-, ClO4-, ArS03 , and C1-C6 AalkylSO3-. 15. A self-assembled helical fibrous nanostructure as described in claim 14 wherein: R1 is methyl; and S' is ArS03. 16. A self-assembled helical fibrous nanostructure as described in claim 11 wherein: R2' and R are both said onium salt. <Desc/Clms Page number 107> 17. A self-assembled helical fibrous nanostructure as described in claim 16 wherein: Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z' and Z2 are both C; R' is C1-C6 alkyl; R2, R3, R4, and R4 are each H; and S' is selected from the group of counter ions consisting of Cl-, Br', I-, BF4-, ClO4-, ArS03 , and C,-C6 alkylSO3-. 18. A self-assembled helical fibrous nanostructure as described in claim 17 wherein: R' is methyl; and S' is ArS03. 19. A self-assembled helical fibrous nanostructure as described in claim 11 wherein: R is said onium salt and R2' is OH. 20. A self-assembled helical fibrous nanostructure as described in claim 19 wherein: Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z' and Z2 are both C; R' is C,-C6 alkyl; R2, R3, R4, and R4 are each H; and S' is selected from the group of counter ions consisting of Cl-, Br-, I', BF4-, ClO4-, ArS03 , and Cj-C alkylSO3-. <Desc/Clms Page number 108> 21. A self-assembled helical fibrous nanostructure as described in claim 20 wherein* R is methyl; and s- is ArSO3-. 22. A self-assembled micellular nanostructure having a spherical shape with a diameter of 20-120 A and incorporating an onium salt of a taxo-diterpenoid- Cn,2-0-aza-arene having an aqueous solubility in excees of 10-3 moles per liter, said onium salt of EMI108.1 said taxo-diterpenoid-Cn, 2-0-aza-arene represented by EMI108.2 the following formula: p10 13 11 9 19 R O '2' RYO 6Ac wherein: Cn is selected from the group consisting of C7 and C2'; RX is selected from the group consisting of Ph and tBuO; R10 is selected from the group consisting of OAc and OH; RY is a C-2 substituent selected from the group consisting of benzyl and the following structures: EMI108.3 and R2' and R7 are each selected from the group consisting of OH and an onium salt of a 2- O-aza-arene, with the proviso that at least <Desc/Clms Page number 109> 2-0-aza-arene, said onium salt of the 2-0aza-arene being selected from the group consisting of onium salt I and onium salt II represented by the following formulas: EMI109.1 wherein: Z' and Z2 are each selected from the group consisting of C and N; Z3 is selected from the group consisting of S and 0; R1 is selected from the group consisting of C1- C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; R2 and R6 are each selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; if Z' is C, then R3 is selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C,-C6 0-alkyl, OH, halogen, and fused aryl; if Z' is N, then R3 is absent; R4 and R are each selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6 0-alkyl, OH, halogen, and fused aryl; and if Z2 is C, then R5 is selected from the group consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6 0-alkyl, OH, halogen, and fused aryl; <Desc/Clms Page number 110> if Z2 is N, then R5 is absent; and s- is a counter ion. 23. A self-assembled helical fibrous nanostructure as described in claim 22 wherein: S' is selected from the group of counter ions consisting of Cl', Br', I', BF4-, ClO4-, ArS03', and C1-C6 alkylSO3-. 24. A self-assembled helical fibrous nanostructure as described in claim 22 wherein: R2' is said onium salt and R is OH. 25. A self-assembled helical fibrous nanostructure as described in claim 24 wherein: RX is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R' is C1-C6 alkyl; R, R3, R4, and R4 are each H; and S' is selected from the group of counter ions consisting of Cl-, Br', I', BF4-, ClO4-, ArS03 , and C1-C6 alkylSO3-. 26. A self-assembled helical fibrous nanostructure as described in claim 25 wherein: R' is methyl; and S' is ArS03-. 27. A self-assembled helical fibrous nanostructure as described in claim 22 wherein: R2' and R7 are both said onium salt. <Desc/Clms Page number 111> 28. A self-assembled helical fibrous nanostructure as described in claim 27 wherein: Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z' and Z2 are both C; R1 is C1-C6 alkyl; R2, R, R4, and R4 are each H; and S' is selected from the group of counter ions consisting of Cl', Br , I', BF4-, ClO4-, ArS03 , and C1-C6 alkylSO3-. 29. A self-assembled helical fibrous nanostructure as described in claim 28 wherein: R1 is methyl; and S' is ArS03-- 30. A self-assembled helical fibrous nanostructure as described in claim 22 wherein: R is said onium salt and R2' is OH . <Desc/Clms Page number 112> 31. A self-assembled helical fibrous nanostructure as described in claim 30 wherein: Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z' and Z2 are both C; R' is C1-C6 alkyl; R2, R3, R4, and R4 are each H; and S' is selected from the group of counter ions consisting of Cl-, Br-, I', BF4-, ClO4-, ArS03 , and C1-C6 alkylSO3-. 32. A self-assembled helical fibrous nanostructure as described in claim 31 wherein: R1 is methyl; and S' is ArS03.

Inventors: GUY RODNEY K, NICOLAOU K C, PITSINOS EMMANUEL, WRASIDLO WOLFGANG, GUY, RODNEY, K., NICOLAOU, K., C., PITSINOS, EMMANUEL, WRASIDLO, WOLFGANG
Applicants: GUY RODNEY K, NICOLAOU K C, PITSINOS EMMANUEL, SCRIPPS RESEARCH INST, WRASIDLO WOLFGANG, GUY, RODNEY, K., NICOLAOU, K., C., PITSINOS, EMMANUEL, THE SCRIPPS RESEARCH INSTITUTE, WRASIDLO, WOLFGANG
Application year: 1995
Main classification: A61K31
IPC codes: A61K31, A61P35, C07D305, C07D405, C07D407, C07D409, C07D411, C07D413, C07D417