WO2001007439A2

WO2001007439A2 - Process for the production of epothiolone b and derivatives as well as intermediate products for this process

Info

Publication number: WO2001007439A2
Application number: PCT/US2000/020064
Authority: WO
Inventors: Johann Mulzer; Harry Martin
Original assignee: Schering Aktiengesellschaft
Priority date: 1999-07-22
Filing date: 2000-07-24
Publication date: 2001-02-01
Also published as: EP1226142A2; JP2003505459A; AU6233600A; NO20020308D0; NO20020308L; WO2001007439A3

Abstract

The present invention is directed to a process for the production of epothilone compounds, the improvement comprising preparing said compounds by cyclization of a compound produced from an intermediate of formula (II) wherein PG is a protecting group.

Description

Process for the Production of Epothiolone B and Derivatives as well as Intermediate Products for this Process

This application claims the benefit of the filing date of U.S. Provisional Application Serial No. 60/145,005, filed July 22 , - ISrS .

This invention relates to a process for the production of epothilone B and derivatives as well as intermediate products for this process.

It is known that the natural substances epothilone A (R = H) and epothilone B (R = methyl) (compound I, DE 195 42 986 Al , DE 41 38 042 C2)

have a fungicidal and cytotoxic effect. According to indications for in vitro activity against mammary and intestinal tumor cell lines, this family of compounds appears especially advantageous for the development of a pharmaceutical agent. Various working groups have successfully endeavored to synthesize these macrocyclic compounds. In this connection, the working groups start from various fragments of the macrocycle to synthesize the desired natural substances.

In any case, diastereomer-pure fragments as starting products and intermediate products are required for a successful epothilone synthesis. Diastereomer purity is often decisive for the action and reliability of a pharmaceutical agent and thus a requirement for its production.

The total synthesis of epothilone A is described by Schinzer et al. in Chem. Eur. J. 1996, 2, No. 11, 1477-1482 and in Angew. Chem. 1997, 109, No. 5, pp. 543-544).

Epothilone derivatives were already described by Hδfle et al. in WO 97/19086. These derivatives were produced starting from natural epothilone A or B.

Another synthesis of epothilone and epothilone derivatives was described by Nicolaou et al. in Angew. Chem. 1997, 109, No. 1/2, pp. 170-172. Nicolaou et al. also described the synthesis of epothilone A and B and several epothilone analogs in Nature, Vol. 387, 1997, pp. 268-272, and the synthesis of epothilone A and its derivatives in J. Am. Chem. Soc, Vol. 119, No. 34, 1997, pp. 7S60-7973 as well as the synthesis of epothilone A and B and several epothilone analogs in J. Am. Chem. Soc, Vol. 119, No. 34, 1997, pp. 7974-7991.

Nicolaou et al. also describe in Angew. Chem. 1997, 109, No. 19, pp. 2181-2187 the production of epothilone A analogs using combinative solid-phase synthesis. Several epothilone B analogs are also described there.

A variable synthesis for the production of epothilone and different types of derivatives is known from WO 99/07692.

Other syntheses are described in PCT Applications WO 99/02514 and WO 99/01124. Finally, the epothilone B-εynthesis that is described by J. Mulzer et al. in Tetrahedron Letters 39 (1998) 8633-8636 can also be mentioned.

Because of the expected lability of the epoxide, in all previous syntheses of epothilone B, the corresponding (Z) -olefin was always epoxidated in the last step, whereby the diaεtereosection is 4:1 to 20:1 of the desired β-isomer. n object of this invention is to indicate a process for the production of epothilone B and epothilone B derivatives, in which the cis-epoxide is introduced at a considerably earlier time via dihydroxylation-monosulfonation of a suitable (E)- olefin, whereby the β-configuraticn of the cis-epoxide is to come

_rcxv_at__c_ Other object are evident ro :f ordinary skill. These objects are achieved by a process using a compound cf

OTBS

ir. v.-hich TΞS stands for a tributylsilyl group. Instead of TBS, another suitable pro ective group can also be another suitable rrctective group can also be used as a starting compound, in v.-hich the epoxy group of the epothilone is already contained, and this epoxy group remains unchanged in all subsequent steps up to the end product. Diagram 3 shows possible derivatizations that allow for the process according to the invention if compound 11 that is to be used and/or next steps 13 or 14 are modified as indicated. This invention therefore extends not only to the process for the production of epothilone B, but also to a process for the production of correspondingly modified derivatives that are derived from modified compounds 11, 13 or 14.

In addition, the invention also relates to the compounds of formulas 5 to 21, which are all new, as well as the correspondingly modified derivatives, which are obtained in the procedures indicated above and in diagram 3.

The selection of protective group (PG) in the 15-hydrcxy group car. be made with cnly routine experimentation. PG should outlast all subsequent reactions up to the macrolactcnization, but in addition should also be removable in the presence of epoxide. The original TΞS-function may not be removed without destroying the substrate; however, after converting the 15-OTBS derivative into the 15-OTΣS analog (TES = triethylεilyl) , any additional synthesis step can be performed easily. Other suitable groups can be routinely determined TBS is preferably replaced by TES at steps 13, 14 or 15; at steps 15 and 17-19, PG is preferably TES.

The epoxide applied early thus was shown as stable under the following reaction conditions:

¹. Reduction (neutral (DIBAH) , ionic (selectride₎ , metallic (Zn)

2. ^Oxidation (osmium tetroxide-sodium periodate₎

³. Bases ⁽fluoride in an aprotic solvent, DMAP, LDA, enolate⁾ . In this case, it is especially surprising that C- and O-anions that were produced intramolecularly at a 1,5-interval to the epoxidic centers do not open the epoxide nucleophilically. 4. Electrophiles (acylation with acid chloride in the

Ya aguchi reaction) . Of all the reagents used, only aqueous acid led to epoxide opening. Apart from the mechanistically valuable finding that does away with the preconception that epoxides are in any case highly reactive synthesis intermediate compounds, the early epoxide introduction alεo has considerable advantages for the production according to the invention of epothilone B and the corresponding derivatives :

The N-oxide formation on thiazole that is observed in the 12 , 13-epoxidation with peracid developε just like the separation of the 12 , 13-epimeric epoxide. No "false" epoxide is produced. The stereoεelection of the aldol reaction iε alεo considerably higher than for the 12 , 13- (Z) -olefin analogs of 21.

The examples that are tied to diagram 3 are used for a more detailed explanation of the invention.

Diagram 1

Synthesis of the Completely Functionalized C15-C7-Fragments of

Epothilone B

1 a : R = B . d.r = 3:1 2a : R = Bn 3a : R = Bπ 1b ^■ R = P.. '3 d.r = 4:1 2b : R = PMB 3b : R = PMB

a R = En b _; - ϊ" 5a : R = Bn 6a : R = Bn 5b : R = PMB 6b : R = PM3

a) 0₃, CH₂C1₂, -78°C, PPh₃; b) isopropenyl-MgBr , THF, -10°C; c) CH₃C(OEt)₃, xylene, 120°C, 12 h; d) DDQ, CH₂C1₂-H₂0, rt; e) Dess-Martin periodinane, CH₂C1₂, rt, 12 h; f) AD-mix-β, tBuOH- H₂0. rt, 20 h; g) Thz-CH₂-PBu₃Cl, KHMDS -78°C then 30°C; h) MsCl, NEt₃, CH₂C1₂, 0°C, 30 min; i) K₂C0₃, MeOH, rt, 45 min; k) DIBAH, CK.C1₂₁ -90°C, 1 h; 1) LiOH (in-situ), (EtO) ₂POCH₂CO (N°) , then aldehyde, Et₂0-THF, rt, 30 min; ) L-selectrides, THF -78°C, 1 h, then HI-IPA, mel, -78°C to 0°C, 4 h; n) DIBAH, CH₂C1_∑, -80°C to -70°C, 2 h.

Bn = benzyl; PMB = p-methoxy-benzyl . All selectrides (cf. Aldrich Chemical Catalog) can be used in the process. L- selectride ( lithium- tri-sec-butylborohydride) is preferred.

Diagram 2

Total Synthesis of Epothilone B (Epoxide Path)

a) LDA, THF, -78°C; b) TrocCl, pyridine, CH₂C1₂, rt; c) i. OsO_A, NMO; ii) NalO ; d) HF-Py, pyridine rt; e) NaCl0₂, NaH₂PO , tBuOH, 2 , 3-dimethyl-2-butene; f) 2 , 4 , 6-trichlorobenzoyl chloride, NEt₃, then DMAP, toluene; g) HF-Py, pyridine rt; h) Zn, NH^Cl, EtOH, reflux, 30 min.

Ketone 16 K.C. Nicolaou et al.: J. Am. Chem. Soc. 1997, 119,

7974-7991

Diagram 3 _Derivatizations

see above

lkyl

Conditions

Products Conditions Products C iiLticns

13a R

13b R

13c R

13d R

13e

13f i3g 13

131 R 13k R 13m R 13n R

vrithcut further elaboration, it is believed that one skilled n. the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific e~_boάiτnents are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

In the foregoing and in the following examples, all te^mperatures are set forth uncorrected in degrees Celsius; and, unless otherwise indicated, all parts and percentages are by weight .

The entire disclosure of all applications, patents and publications, cited above, and U.S. Provisional Application Serial _.c. 60, 145, 005, filed July 22, 1S99 is hereby incorporated _v _τs_fszs^ s . E X A M P L E S

Experimental

64

8.02 g (25 mmol) of alkene 3a iε dissolved in 200 ml of abεolute methylene chloride and mixed with 10 ml of absolute MeOH. After cooling to -78°C, a dried ozone/air mixture is introduced via a gas feed frit until the blue coloring beginε. Air iε allowed to blow through for two more minutes and then quenched by the addition of 19.67 g (75 mmol) of PPh₃ in portions, and it iε allowed to thaw overnight. The solvent is removed as completely as poεεible in Rotavapor, and the remaining εolid reεidue with uεe of preparative column chromatography (hexane/ethyl acetate/ ethylene chloride = 40:1:1^ for separation of PPh₃, then Hx/EE 20:1 to 10:1. 7.90 g (98%) of aldehyde 4a is obtained aε a colorless liquid.

¹H NMR (400 MHz, CDCl₃) δ 9.76 (dd, J = 1.5, 2.0 Hz, 1H) ; 7.34-7.24 ( , 5H) , 4.58 (d, J = 11.8 Hz, 1H) , 4.45 (d, J = 11.8 Hz, 1H) , 4.31 (dt, J = 6.3, 4.5 Hz , 1H) , 3.58-3.51 (m, 1H) , 2.69- 2.62 (m, 1H) , 2.51-2.43 ( , 1H) , 1.14 (d, J = 6.3 Hz, 3H) , 0.83 (ε, 9H) , 0.01 (s, 3H) , 0.00 (s, 3H) .

¹³C NMR (100.6 MHz CDC1₃) S 201.8, 138.4, 128.4, 127.6, 71.0, 68.9, 46.0, 25.7, 17.9, 13.5, -4.7, -4.9.

IR (Film): v_Mχ 2956, 2857, 2712, 1730, 1472, 1255, 1102, 837, 778 cm^"1.

75 ml of isopropenylmagnesium bromide (0.5 M, THF) and 25 ml of absolute THF are cooled to -10°C under Ar atmosphere, and 9.14 g (28.3 mmol) of aldehyde 4a (disεolved in 20 ml of absolute THF) iε slowly (about 20 minutes) added in drops at -10°C. It iε allowed to thaw to 0°C within 45 minuteε (TLC monitoring, Grignard optionally muεt alεo be added) , the reaction mixture is poured into 150 ml of semi-saturated NH₄C1 solution and shaken out after 120 ml of ether is added. The aqueous phase iε extracted twice more with ether (100 ml) . The combined organic phases are dried (MgSO , and the solvent iε removed in Rotavapor. Preparative column chro atography (Hx/EE = 10:1) yieldε 9.48 g (92%) of allyl alcohol 5a (diaεtereomer mixture) aε a colorless, viεcouε liquid.

Diaεtereomer 1

¹H NMR (400 MHz, CDC1₃) <S 7.34-7.24 (m, 5H) , 4.98 (s, 1H) , 4.80 (ε, 1H) , 4.60 (d, J = 12.0 Hz, 1H) , 4.53 (d, J = 12.0 Hz, 1H) , 4.17 (d_br, J = 9.5 Hz, 1H) , 4.02 (dt, J = 7.4, 4.5 Hz , 1H) , 3.61-3.54 ( , 1H) , 2.71 (d_bp, J = 3.5 Hz, 1H) , 1.84-1.75 (m, 1H) , 1.72 (ε, 3H) , 1.65-1.57 (m, 1H) , 1.14 (d, J = 6.5 Hz, 3H) , 0.86 (ε, 9H) , 0.05 (ε, 3H) , -0.02 (ε, 3H) . Diastereomer 2

'H NMR (400 MHz, CDCl₃) δ 7.34-7.24 (m, 5H) , 4.99 (ε, 1H) , 4.83 (s, 1H) , 4.60 (d, J = 12.0 Hz, 1H) , 4.47 (d, J = 12.0 Hz, 1H) , 4.20 (dd, J = 8.0, 3.5 Hz, 1H) , 3.90 (dt, J = 8.5, 4.3 Hz , 1H) , 3.47 (dq, J = 6.4, 4.3 Hz, 1H) , 2.97 (s_bp, 1H) , 1.86 (dt, J = 14.2, 4.1 Hz, 1H) , 1.67 (s, 3H) , 1.57 (dt, J = 14.6, 8.5 Hz, 1H) _. 1.10 (d, J = 6.4 Hz, 3H) , 0.80 (s, 9H)

8.02 g (22 mmol) of allyl alcohol 5a (diaεtereomer mixture) iε disεolved in 120 ml of absolute xylene and mixed with 32 ml (176 mmol) of triethylorthoacetate and 4 dropε of propionic acid. Kith a light Ar stream through a thin capillary, it is stirred for 16-20 hourε at 120°C, whereby the ethanol that is produced is distilled off via a εmall distillation _{appara us} . _After the reaction iε completed (TLC monitoring) , the solvent iε distilled off in Rotavapor (30 mbar, 50°C, then complete vacuum) , and the crude product iε purified by column chromatography (Hx/EE = 40:1 gradient 20:1). 8.77 g (92%) of 6a iε obtained-as a colorless liquid.

¹H KM (400 MHz, CDC1-.) δ 7.35-7.22 (m, 5H) ; 5.20 (t, J = 6.4 Hz, 1H) ; 4.57 (d, J = 12 Hz, 1H) , 4.48 (d, J = 12 Hz, 1H) , 4.11 (q, J - 7.2, 2H) , 3.68 (m, J = 4 Hz, 1H) , 3.46 (dt, J = 6.3, 4.5 Hz, 1H) , 2.40-2.34 (m, 1H) , 2.32-2.24 (m, 2H) , 2.14-2.04 ( , 1H) , 1.60 (ε, 3H) , 1.23 (t, J = 7.2 Hz, 2H) , 1.11 (d, J = 6.4 Hz, 3H) , 0.84 (ε, 9H) , -0.02 (ε, 3H) , -0.05 (s, 3H) .

¹³C NMR (100.6 MHZ CDC1₃) δ 173.5, 139.1, 134.5, 128.2, 127.5, 127.4, 124.0, 122.4, 77.3, 74.0, 71.0, 60.2, 34.8, 33.1, 30.1, 25.8, 18.0, 16.1, 14.2, 14.0, -4.60, -4.61.

IR (Film) : v_maχ 2956, 2930, 2887, 2857, 1737, 1472, 1462, 1454, 1370, 1300, 1255, 1155, 1098, 939, 836, 776, 737 cm^"1.

HRKS (El) Cld. for C₂₅H₄₂0 Si 434.2852, Fnd. 434.2845

[ ]²⁰ _D - -3.9 (c = 1.2. CHC1₃)

S12 g (2.10 mmol) of 6a iε dissolved in 40 ml of methylene chloride and 2 ml of water, mixed with 2.86 g (12.6 mmol) of DDQ and εtirred vigorouεly for exactly 3 hourε at room temperature. The reaction mixture iε diluted with 100 ml of ether and washed with NaHC0₃ (aqueouε, εaturated) (2x 40 ml) . The combined aqueouε phaεeε are diluted with 80 ml of water and extracted with ether (2x 50 ml) . The combined organic phaεeε are waεhed with brine (50 ml) , dried (MgSO , and the solvent is removed in Rotavapor. Preparative column chromatography (Hx/EE = 10:1) yields 575 mg (80%) of alcohol 7 as a colorlesε liquid.

¹H NMR (400 MHz, CDCl₃) δ 5.16 (dt, J = 7.3, 1.0 Hz, IH) , 4.09 (q, J = 7.07 Hz , 2H) , 3.56 (m, IH) , 3.42 (dt, J = 7.2, 4.4 Hz, IH) , 2.40-2.26 (m, 5H) , 2.15-2.07 (m, 2H) , 1.61 (m, 3H) , 1.23 (t, J = 7.0 Hz, 3H) , 1.09 (d, J = 6.5 Hz, 3H) , 0.88 (ε, 9H) , 0.06 (ε, 6H) .

¹³C NMR (100.6 MHZ CDC1₃) <S 173.7, 136.1, 120.9, 77.0, 69.0, 60.7, 35.2, 33.5, 33.0, 26.2, 20.3, 18.5, 16.6, 14.6, -3.8, -4.3.

IR (Film) : v_MX 3513 (br) , 2932, 2874, 2855, 1738, 1463, 1372, 1255, 1157, 1088, 836, 777 cm^"1.

HRMS (El) Cld. for (M⁺ -C₄H_?) 287.168, Fnd . 287.168 ± 5 ppm

[α]²⁰ _D = +20.2 (C = 1.08, CHC1₃)

1.030 g (2.99 mmol) of alcohol 7 iε diεεolved in 50 ml of absolute methylene chloride and mixed with 2.5 ml of absolute pyridine and 2.54 g (6.0 mmol) of Desε-Martin periodinane and εtirred overnight at RT (about 18 hours) . For working-up, it is mixed with 25 ml of sodium thioεulfate εolution (20%, aqueous), and it iε εtirred vigorouεly for 15 minuteε. The phases are separated, and the aqueouε phase is extracted with methylene chloride (2 x 25 ml) . The combined organic phaseε are dried (MgS0₄) , and the solvent is removed in Rotavapor. Preparative column chromatography (Hx/EE = 10:1) yieldε 945 mg (92%) of ketone 8 as a colorleεε liquid.

¹H NMR (400 MHz, CDC1₃) δ 5.15 (t, IH, J = 6.8 Hz , IH) ; 4.10 (q, J = 7.0 Hz, 2H) , 3.97 (dd, J = 6.5, 5.5 Hz, IH) , 2.40- 2.20 ( , 6H) , 2.12 (s, 3H) , 1.59 (ε, 3H) , 1.23 (t, J = 7.0 Hz, IH) , 0.89 (ε, 9H) , 0.03 (ε, 3H) , 0.02 (ε, 3H) .

¹³C NMR (100.6 MHZ CDC1₃) δ 212.5, 173.7, 137.0, 119.7, 79.2, 60.7, 35.1, 33.9, 33.7, 33.4, 26.1, 25.9, 18.5, 16.6, 14.6, -4.5, -4.6.

IR (Film): v_maχ 2932, 2875, 2858, 1737, 1351, 1256, 1203, 1160, 1137, 1102, 959, 927, 899, 839, 779 cm^"1.

HRMS (El) Cld. for (M⁺-CH₃) 327.1992. Fnd. 327.199 ± 0.0016

[ Q ] ²° = - 19 . 4 ( c = 1 . 05 , CHCl, )

a) 830 mg (2.42 mmol) of 8 is diεsolved in 30 ml of _tBuOH- H₂0 (1:1) and mixed with 1.5 g of NaHC0₃, 5.0 g of AD-Mix-β and 230 g of methaneεulfonamide. After 36-48 hourε at room temperature while being εtirred vigorouεly, it iε quenched by adding 5 g of εodium εulfite (10 more inuteε of εtirring) . After 25 ml of H.,0 and 100 ml of methylene chloride are added, the phaεeε are εeparated, and the aqueouε phaεe iε extracted with methylene chloride (3 x 30 ml) . The combined organic phases are dried (MgSO , and the solvent is removed in Rotavapor. Preparative column chromatography (Hx/EE = 2:1, R. _(3-1. = 0.09 to 0.29) yieldε 942 mg of product mixture 8a.

IR (Film): v_maχ 3432, 2933, 2858, 1736, 1377, 1256, 1229, 1095, 837, 778 cm^'1.

b) 1.82 g of Wittig salt (5.2 mmol) iε diεεolved in 30 ml of absolute THF under Ar atmoεphere, cooled to -78°C, and 10.4 ml of KHKDS (0.5 M, toluene) iε added in dropε and stirred for 45 minutes at -78°C. Then, 920 mg of 8a (dissolved in 2.5 ml of THF⁾ is added smoothly in drops. It is allowed to stir for 5 more minutes

at -78°C, the cooling bath iε then quickly exchanged for an approximately 40°C water bath and allowed to thaw. After 5 minuteε, the bath is removed, and it iε allowed to stir for another 5 minuteε before being quenched with 50 ml of NH^Cl (saturated, aqueous) and 75 ml of ether. The aqueous phase is extracted with ether (2x 30 ml) . The combined organic phases are dried (MgSO , and the solvent is removed in Rotavapor. Preparative column chromatography (Hx/EE = 2:1) yields 632 mg (61% on both εynthesiε εtepε) of alkene 9 as a colorless, viεcouε liquid.

¹H NMR (400 MHz, CDCl₃) δ 6.93 (ε, IH) , 6.47 (ε, IH) , 4.41 (dd, J = 8.5, 4.5 Hz, IH) , 3.67 (d, J = 10.0 Hz, IH) , 3.45 (s, IH) , 2.71-2.61 (m, IH) , 2.68 (ε, 3H) , 2.66-2.46 (m, IH) , 2.37- 2.28 (m, IH) , 1.99 (ε, 3H) , 1.80-1.66 (m, 2H) , 1.70 (dd, J = 14.3, 4.8 Hz, IH) , 1.34 (ε, 3H) , 0.88 (ε, 9H) , 0.10 (ε, 3H) , 0.01 (ε, 3H) .

¹²C NMR (100.6 MHZ CDC1₃) <S 177.6, 165.1, 152.9, 141.4, 120.4, 116.3, 88.0, 79.9, 76.7, 37.1, 30.9, 29.9, 26.2, 23.6, 19.6, 18.4, 14.1, -4.0, -4.8.

IR (Film): v_maχ 3469, 2955, 2943, 2930, 2910, 2892, 2856, 1768, 1656, 1505, 1462, 1386, 1252, 1066, 838 cm^'1.

HRMS (El) Cld. for C21H35N04SiS 425.2056 Fnd . 425,2060 ± 0.0025

[α]²⁰ _D = -27.1 (c = 1.0, CHC1₃)

12.7

313 mg (0.737 mmol) of alcohol 9 iε diεεolved under Ar atmoεphere in 5 ml of abεolute methylene chloride and mixed at 0°C with 0.31 ml of abεolute triethylamine and 0.09 ml of methaneεulfonic acid chloride. After 30 minutes, it is quenched with 10 ml of NaHC0₃ (aqueouε, εaturated) , the phaεeε are εeparated, and the aqueouε phaεe iε extracted with methylene chloride (3x 10 ml) . The combined organic phaεeε are dried (MgS0₄) , and the solvent iε removed in Rotavapor. Preparative column chromatography (Hx/EE = 4:1) yields 283 mg (76%) of meεylate 10 aε a strongly viεcouε, yellowish liquid.

¹H NMR (400 MHz, CDC1₃) δ 6.96 (ε, IH) , 6.56 (ε, IH) , 4.57 (dd, J = 8.0, 2.5 Hz, IH) , 4.41 (dd, J = 8.8, 4.8 Hz, IH) , 3.13 (ε, 3H) , 2.68 (ε, 3H) , 2.60-2.53 (m, 2H) , 2.02 (d, J = 1.0 Hz, 3H) , 2.07-1.81 (m, 4H) , 1.42 (ε, 3H) , 0.87 (ε, 9H) , 0.08 (ε, 3H) , 0.02 (ε, 3H) .

¹³C NMR (100.6 MHz CDC1₃) δ 175.5, 165.0, 153.2, 138.7, 122.1, 117.2, 86.5, 83.8, 75.7, 39.6, 38.0, 31.2, 28.8, 26.2, 21.7, 19.7, 18.5, 13.1, -4.3, -4.6.

256 g (0.508 mmol) of meεylate 10 iε diεεolved in 10 ml of abεolute methanol and mixed with finely pulverized potaεsium carbonate. After 45 minutes at room temperature, it is diluted with 30 ml of ether and then filtered. Saturated NH^Cl solution (15 ml) iε added to the filtrate until two clear phaεes form. The phaεeε are εeparated, and the aqueous phaεe iε extracted with ether (3x 10 ml) . The combined organic phaεeε are dried (MgSO , and the εolvent iε removed in Rotavapor. Preparative column chromatography (Hx/EE = 5:1) yields 202 mg (90%) of epoxide 11 as a colorless, viεcouε liquid.

¹H NMR (400 MHz, CDC1₃) <S 6.91 (s, IH) , 6.49 (s, IH) , 4.32 (dd, J = 9.0, 3.5 Hz, IH) , 3.65 (s, 3H) , 2.89 (dd, J = 7.0, 4.5 Hz, IH) , 2.68 (ε, 3H) , 2.46-2.40 (m, 2H) , 1.99 (d, J = 1.5 Hz, IH) , 1.94-1.77 (m, 3H) , 1.63-1.55 (m, IH) , 1.26 (ε, 3H) , 0.88 (s, 9H) , 0.07 (ε, 3H) , 0.01 (s, 3H) .

¹³C NMR (100.6 MHz CDC1₃) δ 173.6, 164.9, 153.4, 142.4, 119.3, 115.8, 76.7, 62.8, 60.4, 52.1, 36.3, 30.5, 28.6, 26.3, 22.5, 19.6, 18.6, 14.4, -4.2, -4.7.

IR (Film) : v_maχ 2955, 2874, 2857, 1740, 1656, 1505, 1439, 1380, 1312, 1230, 1180, 1077, 835, 778 cm^"1.

HRMS (El) Cld. for C_∑2H₃₇N0₄SiS 439.2213 Fnd. 439.221 ± 0.0025.

[ ]²⁰ = -12.6 (c = 1.04, CHC1,)

176 mg (0.40 mmol) of epoxide 11 iε dissolved in 6 ml of abεolute MC under argon and εlowly (about 5 minuteε) mixed with 0.29 ml (0.44 mmol) of DIBAH (1.5M toluene) ^' at -95°C. It iε allowed to come to -85°C within 45 minuteε, mixed with 0.15 ml of NH₄C1 solution and 10 ml of ether while being εtirred vigorouεly, and it iε allowed to thaw quickly. After sufficient MgS0₄ is added, it is allowed to stir for 1-2 hours and filtered off. After the solvent is removed in Rotavapor, the crude aldehyde is purified by column chromatography (Hx/EE = 3:1), and 140 mg (85%) of aldehyde 11a is obtained as a colorless oil.

IR (Film): v_Mχ 2930, 2781, 1727, 1677, 1076, 919, 836.

4 ml of abεolute ether at 0°C and 0.32 ml of nBuLi (1.6M, hexane, 0.51 mmol) are introduced under Ar atmosphere and mixed with 0.36 ml of THF/H-,0 (19:1, 1 mmol of H-,0) . After 5 minutes, 193 g of phosphonate (0.49 mmol, dissolved in 1 ml of THF) is added in drops, and after another 10 minutes, 135 mg (0.33 mmol, dissolved in 1 ml of THF) of aldehyde 11a is added in dropε. The cooling bath is removed, and after 30 minutes at room temperature, it iε quenched with 4 ml of saturated NH₄C1 solution, the phases are εeparated, and the aqueouε phaεe iε extracted with ether (2x 5 ml) . The combined organic phaεeε are dried (MgS0₄) , and the solvent is removed in Rotavapor. Preparative column chromatography (Hx/EE = 3:1) yields 194 mg (91%) of enoylsultam 12 aε a viscous oil, which solidifies like a foam after freezing-out.

Η NMR (400 MHz, CDC1₃) δ 7.03 (dt, J = 14.8, 7.3 Hz, IH) , 6.91 (s, IH) , 6.55 (d, J = 14.8 Hz, IH) , 6.49 (s, IH) , 4.32 (dd, J = 8.8, 3.8 Hz, IH) , 3.90 (dd, J = 7.3, 5.3 Hz, IH) , 3.48 (d, J = 13.8 Hz, IH) ; 3.40 (d, J = 13, 8 Hz, IH) , 2.89 (dd, J = 7.5, 4.5 Hz, IH) , 2.68 (ε, 3H) , 2.42-2.33 (m, 2H) , 2.13-2.02 (m, 2H) , 1.99 (ε, 3H) , 1.95-1.81 (m, 5H) , 1.71-1.51 (m, 4H) , 1.27 (s, 3H) , 1.14 (ε, 3H) , 0.95 (ε, 3H) , 0.88 (ε, 9H) , 0.07 (ε, 3H) , 0.01 (s, 3H) . C NMR (100.6 MHz CDC1₃) δ 164.8, 164.3, 153.4, 149.9, 142.4, 121.6, 119.3, 115.8, 76.8, 65.5, 62.6, 60.7, 53.5, 50.7, 48.8, 48.2, 45.1, 38.9, 36.2, 33.3, 31.9, 28.8, 26.9, 26.2, 22.6, 21.2, 20.3, 19.6, 18.6, 14.4, -4.2, -4.7.

172 mg (0.265 mmol) of sultam 12 iε diεεolved in 5 ml of abεolute THF and mixed at -95°C with 0.32 ml of L-εelectrides (1.0 M, THF, 0.32 mmol) and heated within 45 minuteε to -40°C. After another 15 minuteε, it is cooled to -78°C, and after 0.12 ml of HMPA iε added, it iε mixed with 0.132 ml of Mel. Then, it iε heated within 3 hourε to 0°C, and after another 2 hourε at 0°C, it iε quenched by adding 5 ml of NH_ACl and 10 ml of ether. The phaεeε are εeparated, and the aqueouε phase is extracted with ether (2x 5 ml) . The combined organic phaseε are dried (MgS0₄) , and the solvent is removed in Rotavapor. Preparative column chromatography (Hx/EE = 3:1) yieldε 90 mg of 13 and 85 mg of 14.

Compound 13

¹H NMR (400 MHz, CDC1₃) <S 6.88 (ε, IH) , 6.47 (ε, IH) , 4.29 (dd, J = 9.0, 3.5 Hz, IH) , 3.85 (t, J = 6.3 Hz, IH) , 3.44 (d, J = 13.5 Hz, IH) , 3.37 (d, J = 3.5 Hz, IH) , 3.07-3.97 (m, IH) , 2.83 (dd, J = 8.0, 4.0 Hz, IH) , 2.66 (s, 3H) , 2.02-1.98 (m, 2H) , 1.98 (d, J = 1.0 Hz, 3H) , 1.92-1.68 (m, 5H) , 1.57-1.26 (m, 8H) , 1.22 (ε, 3H) , 1.16 (d, J = 7.0 Hz, 3H) , 1.11 (s, 3H) , 0.92 (s, 3H) , 0.86 (s, 9H) , 0.05 (ε, 3H) , -0.01 (ε, 3H) .

Compound 14

^ NMR (400 MHz, CDC1₃) «5 6.91 (ε, IH) , 6.49 (ε, IH) , 4.31 (dd, J = 9.0, 3.5 Hz, IH) , 3.83, (dd, J = 7.3, 5.3 Hz, IH) , 3.45 (d, J = 13.6 Hz, IH) , 3.39 (d, J = 13.6 Hz, IH) , 2.86 (dd, J = 7.8, 4.3 Hz, IH) , 2.75-2.63 (m, 2H) , 2.68 (ε, 3H) , 2.12-2.03 (m, 2H) , 2.00 (d, J = 1.0 Hz, 3H) , 1.93-1.80 (m, 4H) , 1.72-1.30 (m, 9H) , 1.24 (ε, 3H) , 1.12 (ε, 3H) , 0.93 (s, 3H) , 0.88 (ε, 9H) , 0.07 (ε, 3H) , 0.01 (s, 3H) .

50 mg (0.075 mmol) of sultam 13 is dissolved under argon in 2.5 ml of absolute methylene chloride and mixed slowly with 0.083 ml (0.083 mmol) of DIBAH (1.0 M, toluene) at -95°C. It iε allowed to come to -75°C within 2-2.5 hours while being εtirred vigorously with 0.05 ml of MeOH and 0.1 ml of NH^Cl solution, as well aε 5 ml of ether, and it iε allowed to thaw quickly. After εufficient MgS0₄ is added, it iε allowed to εtir for about 1 hour (even overnight) and filtered off. After the εolvent iε removed in Rotavapor, the crude aldehyde iε purified by column chromatography (Hx/EE = 3:1), and 24 mg (71%) of aldehyde 15 iε obtained aε a waxy εolid.

¹H NMR (400 MHz, CDC1₃) 6 9.58 (d, J = 5.0 Hz, IH) , 6.90 (ε, IH) , 6.49 (ε, IH) , 4.31 (dd, J = 9.0, 4.0 Hz, IH) , 2.87 (dd, J = 7.0, 4.5 Hz, IH) , 2.68 (ε, 3H) , 2.36-2.26 ( , IH) , 2.00 (ε, 3H) , 1.92-1.84 (m, IH) , 1.74-1.64 (m, IH) , 1.60-1.52 (m, IH) , 1.50-1.32 (m, 5H) , 1.26 (ε, 3H) , 1.08 (d, J = 7.0 Hz, 3H) , 0.88 (ε, 9H) , 0.07 (ε, 3H) , 0.01 (ε, 3H) . ¹³C NMR (100.6 MHz CDC1-.) δ 205.2, 164.9, 153.4, 142.5, 119.2, 115.8, 76.7, 62.4, 61.1, 46.7, 36.4, 33.6, 31.0, 26.2, 23.3, 22.7, 19.6, 18.6, 14.4, 13.7, -4.2, -4.7.

Aldol Reaction

Example: R² = TES, R³ = TBS

1.75 ml (1.23 mmol) of iεopropylamine iε diεεolved in 4 ml of absolute THF and mixed at -45°C with 0.75 ml of nBuLi (1.6 M in hexane; 1.20 mmol), heated to 0°C after 5 minuteε and εtirred for 20 minuteε at 0°C. After cooling to -78°C, 345 mg (1.21 mmol) of ketone 16 (diεεolved in 1 ml of THF) iε added in dropε within 2 minuteε, and the reaction mixture iε heated to -40°C within 30 minuteε. It iε again cooled to -78°C, and 532 mg (1.18 mmol) of aldehyde 15 (diεεolved in 1.5 ml of THF) is added in dropε within 2-3 minuteε. After 15 minuteε at -78°C, it is quenched with 4 ml of εaturated NH₄C1 εolution while being εtirred vigorouεly (initially εlowly then quickly added) , 6 ml of ether iε added, and the cooling bath iε replaced by a water bath. After thawing, εome water iε added, and the phaεeε are εeparated after εhaking out. Extraction with ether, drying (MgSO_A) , removal of the εolvent and subsequent column chromatography (hexane/ethyl acetate 10:1 → 5:1) yield 668 mg (77%) of desired main iεomer 17 aε a colorleεε, viscouε liquid.

Compound 17 (R² = TES, R³ = TBS)

¹H NMR (400 MHz, CDCl₃) δ = 6.93 (s, IH) , 6.51 (ε, IH) , 5.72 (ddt, J = 17.1, 10.0, 7.0 Hz, IH) , 4.99 (m, 2H) , 4.33 (dd, J = 9.0, 3.5 Hz, IH) , 3.92 (d, J = 6.3, 4.3 Hz, IH) , 3.49 (s, IH) , 3.31 (d, J = 9.0 Hz, IH) , 3.25 (q, J = 7.0 Hz, IH) , 2.88 (dd, J = 7.5, 4.0 Hz, IH) , 2.70 (ε, 3H) , 2.24-2.06 (m, 2H) , 2.01 (d, J = 1.0 Hz, 3H) , 1.95-1.87 (m, IH) , 1.82-1.72 (m, IH) , 1.63-1.45 (m, 5H) , 1.41-1.30 ( , IH) , 1.28 (ε, 3H) , 1.18 (ε, 3H) , 1.20-1.05 (m, IH) , 1.12 (ε, 3H) , 1.03 (d, J = 7.0 Hz , 3H) , 0.90 (ε, 9H) , 0.89 (ε, 9H) , 0.83 (d, J = 7.0 Hz, 3H) , 0.09 (s, 3H) , 0.07 (ε, 3H) , 0.06 (ε, 3H) , 0.03 (ε, 3H) , ¹³C NMR (100.6 MHz, CDCl₃) : δ = 222.5, 164.4, 153.1, 142.2, 136.3, 118.8, 116.7, 115.3, 76.5,

76.4, 74.9, 62.1, 61.1, 54.3, 41.1, 39.6, 36.0, 35.6, 33.4, 33.1,

26.05, 25.9, 23.4, 22.7, 22.4, 19.4, 19.2, 18.24, 18.21, 15.3, 14.0, 9.7, -3.5, -4.90, -4.6, -5.1.

7-OH Protection

Example: R² = TES, R³ = TBS, R⁴ = Troc

200 mg (0.27 mmol) of aldol 17 iε diεεolved in 10 ml of methylene chloride and 5 ml of pyridine and mixed at 20°C (water bath) with 0.5 ml (2.4 mmol) of chloroformic acid-2,2,2- trichloroethyl ester and stirred for 30-45 minutes at room temperature. For working-up, the reaction mixture iε εhaken out with 50 ml of εaturated NaHC0₃ εolution and 40 ml of methylene chloride. Extraction with methylene chloride, drying (MgS0₄) , removal of the solvent and εubεequent column chromatography (hexane/ethyl acetate 10:1) yield 231 mg (94%) of deεired product 18 aε a colorleεε oil.

Compound 18 (R² = TES, R³ = TBS, R⁴ = Troc)

¹H NMR (400 MHz, CDC1₃) δ = 6.92 (ε, IH) , 6.51 (ε, IH) , 5.79 (ddt, J = 16.6, 10.5, 7.0 Hz, IH) , 5.05-4.97 (m, 2H) , 4.85 (d, J = 12.1 Hz, IH) , 4.81 (dd, J = 7.0, 4.5 Hz, IH) , 4.69 (d, J = 12.1 Hz, IH) , 4.33 (dd, J = 9.0, 3.5 Hz, IH) , 3.75 (dd, J = 6.5, 4.0 Hz, IH) , 3.44 (m, IH) , 2.88 (dd, J = 7.5, 4.0 Hz, IH) , 2.70 (ε, 3H) , 2.28-2.19 (m, IH) , 2.06-1.95 (m, IH) , 2.02 (ε, 3H) , 1.92-1.84 (m, IH) , 1.75-1.65 ( , IH) , 1.57-1.41 (m, 5H) , 1.33- 1.11 (m, 2H) , 1.30 (ε, 3H) , 1.27 (ε, 3H) , 1.21-1.12 (m, IH) , 1.07 (d, J = 7.0 Hz, 3H) , 1.04 (ε, 3H) , 0.96 (d, J = 7.0 Hz, 3H) , 0.91 (ε, 9H) , 0.89 (ε, 9H) , 0.09 (ε, 3H) , 0.07 (ε, 3H) , 0.06 (ε, 3H) , 0.03 (s, 3H) , ¹³C NMR (100.6 MHz, CDC1₃) : <5 = 215.6, 164.4, 154.2, 153.1, 142.1, 136.4, 118.9, 116.6, 115.4, 94.8, 82.9, 78.1, 76.6, 76.4, 62.1, 60.8, 54.0, 42.3, 39.4, 36.0, 35.0, 33.3, 31.8, 26.1, 25.9, 24.0, 22.7, 22.3, 20.0, 19.2, 18.23, 18.21, 16.1, 14.0, 11.5, -3.6, -3.9, -4.6, -5.1. IR (film) ϋ^ = 2956, 2930, 2858, 1760, 1699, 1472, 1384, 1251, 1081, 992, 928, 836, 777, 732. Rotation: [α]²⁰ _D = -30 (c = 1.4, CH₂Cl₂) . Analysis Cld. for C₄₃H₇₄Cl₃N0₇SSi₂ (911,65): C 56.65, H 8.18, N 1.54, Fnd.: C 56.54 H 8.18 N 1.47.

Dihydroxylation/Glycol Cleavage

Example: R² = TES, R³ = TBS, R^c = Troc

148 g (0.162 mmol) of alkene 18 iε dissolved in 8 ml of TKF-tBuOH (1:1) and mixed with 2 mg of Os04 (5 mol%) and 0.89 ml of KMO (0.2 M in H₂0, 0.178 mmol). After 16 hourε of εtirring at 25°C, it iε shaken out vigorously and for a long time with 10 ml of Na-.S-.0₃ (10% in H_∑0) and 15 ml of methylene chloride. The phases are separated, and the aqueouε phaεe iε extracted three more times with methylene chloride. Drying (MgS0₄) , removal of the εolvent and filtration via a εhort silica gel column (hexane/ethyl acetate 1:1) yields 131 mg (86%) of diol aε an iεoroer mixture, which iε further uεed without additional purification.

131 mg (0.139 mmol) of diol iε diεεolved in 15 ml of ethanol and 3 ml of H-.0 and mixed with 90 mg (0.420 mmol) of NaI0₄. After three hours of stirring at 25°C, it is shaken out with 30 ml of semi-saturated NaHC0₃ solution and 30 ml of ether, and the phases are separated. Extraction with ether, drying (MgS0₄) , removal of the εolvent and εubεequent column chromatography (Hexane/ethyl acetate 10:1 → 5:1) yield 115 mg (78% in terms of alkene) of aldehyde 19 aε a colorleεε oil.

Compound 19 (R² = TES, R³' = TBS, R⁴ = Troc)

¹H NMR (400 MHz, CDC1₃) : <5 = 9.73 (dd, J = 2.0, 1.0 Hz, IH) , 6.92 (ε, IH) , 6.50 (ε, IH) , 4.83 (d, J = 12.0 Hz, IH) , 4.74 (dd, J = 7.5, 4.0 HZ, IH) , 4.67 (d, J = 11.8 Hz, IH) , 4.36-4.29 ( , 2H) , 3.49-3.38 (m, IH) , 2.88 (dd, J = 7.5, 4.0 Hz, IH) , 2.70 (ε, 3H) , 2.67 (ddd, J = 17.5, 4.5, 1.0 Hz, IH) , 2.39 (ddd, J =

17.5, 5.5, 2.0 Hz, IH) , 2.39 (s, 3H) , 1.88 (ddd, J = 13.9, 9.4, 4.0 Hz, IH) , 1.76-1.66 ( , IH) , 1.60-1.40 (m, 5H) , 1.35-1.10, (m, 2H) , 1.33 (s, 3H) , 1.26 (s, 3H) , 1.20 (d, J = 6.5 Hz, IH) , 1.02 (s, 3H) , 0.97 (d, J = 6.5 Hz, 3H) , 0.90 (s, 9H) , 0.86 (ε, 9H) , 0.11 (ε, 3H) , 0.09 (ε, 3H) , 0.03 (ε, 3H) , 0.02 (ε, 3H) , ¹³C NMR (100.6 MHz, CDC1₃) : <5 = 215.4, 200.2, 164.2, 154.2, 153.0, 142.1, 118.9, 115.4, 94.7, 82.3, 76.65, 76.3, 72.3, 62.1, 60.8, 53.4, 49.3, 42.2, 35.9, 34.9, 3.33, 31.9, 25.87, 25.85, 23.0,

22.6, 22.3, 20.0, 19.2, 18.2, 18.1, 15.9, 14.0, 11.1, -4.4, -4.5, -4.6, -5.1. IR (Film) fi_Mj( = 2956, 2930, 2885, 2857, 1759, 1726, 1699, 1472, 1384, 1361, 1252, 1082, 992, 927, 836, 777, 734. Rotation: [c.]²⁰ _D = -42.7 (c = 1.2, CHC1₃) 15-O-Protection Removal

Example: R₂ = TES, R³ = TBS, K¹ = Troc

68 mg (0.074 mmol) of 19 iε diεεolved in a polypropylene reaction vesεel (with a cover) in 2.5 ml of abεolute THF and mixed with 2.5 ml of a standard solution of HF-pyridine (produced from: 5 ml of HF-pyridine, 15 ml of pyridine and 10 ml of THF) that is buffered with pyridine. After 30 minuteε, the reaction iε completed. For working-up, 80 ml of εaturated NaHC0₃ εolution iε introduced, and the reaction mixture iε carefully added (plastic syringe) while being εtirred vigorouεly. After extraction with ether (4 timeε 25 ml) and drying (MgS0₄) , the εolvent iε removed in Rotavapor, whereby the pyridine iε removed by repeated εpinning-in with toluene. The reεidue iε put on a column with deactivated εilica gel (hexane/ethyl acetate 2 : 1 → 1:1), and 51 mg (92%) of product 20 iε obtained aε a pale yellow, viεcouε oil.

Compound 20 (R³ = TBS, R = Troc)

Η NMR (400 MHz, CDC1₃) <S = 9.74 (ε, IH) , 6.95 (ε, IH) , 6.60 (ε, IH) , 4.84 (d, J = 12.0 Hz, IH) , 4.75 (dd, J = 7.8, 4.3 Hz, IH) , 4.68 (d, J = 12.0 Hz , IH) , 4.41-4.35 (m, IH) , 4.34 (t, J = 4.8 Hz, IH) ; 3.49-3.41 (m, IH) , 2.96 (dd, J = 8.0, 4.0 Hz, IH) , 2.96 (ε, 3H) , 2.67 (dd, J = 17.5 Hz, 4.0 Hz , IH) , 2.40 (ddd, J =

17.5, 5.5, 2.0 Hz, IH) , 2.09 (d, J = 3.5 Hz, IH) , 2.06 (ε, 3H) , 1.94 (ddd, J = 14.6, 8.5, 4.0 Hz, IH) , 1.78-1.68 (m, IH) , 1.67 (ddd, J = 14.0, 8.0, 4.0 Hz, IH) , 1.58-1.41 (m, 4H) , 1.35-1.25 (m, IH) , 1.34 (ε, 3H) , 1.28 (ε, 3H) , 1.07 (d, J = 6.5 Hz, 3H) , 1.04 (ε, 3H) , 0.98 (d, J = 7.0 Hz , 3H) , 0.87 (ε, 9H) , 0.11 (ε, 3H) , 0.03 (ε, 3H) , ¹³C NMR (100.6 MHz, CDCl₃) : δ = 215.4, 200.5, 164.6, 154.2, 152.7, 141.7, 118.9, 94.7, 82.3, 76.7, 75.4, 72.2, 61.8, 60.7, 53.4, 49.3, 42.2, 34.9, 34.1, 33.2, 32.0, 25.9, 23.0,

22.6, 22.1, 20.0, 19.2, 18.1, 15.9, 14.5, 11.2, -4.4, -4.5.

Pinnick-Oxidation/Macrolactonization

Example: R³ = TBS, R = Troc

54 mg (0.068 mmol) of aldehyde 20 iε diεεolved in 2.5 ml of tBuOH and 2.5 ml of 2 , 3-dimethyl-2-butene and mixed with a εolution of 30 mg of NaCl0₂ and 30 mg of NaH₂P0₄ in 0.5 ml of H₂0 and εtirred for 45 minuteε at room temperature. For working-up, the reaction mixture iε mixed with 20 ml of εemi-εaturated NH₄Cl εolution and extracted with methylene chloride (4 x 10 ml) . After drying, the solvent is removed, and crude acid 21 (51 mg) that iε thuε obtained iε uεed directly in the next reaction.

51 g (0.062 mmol) of crude acid 21 iε diεεolved in 1.5 ml of absolute THF and mixed at 0°C with 57 μl (0.36 mmol) of triethylamine and 43 μl (0.24 mmol) of 2 , 4 , 6-trichlorobenzoyl chloride and εtirred for 20 minuteε at room temperature. The active eεter that is thus produced is εlowly added in dropε (15 minuteε) to a solution of 80 mg (0.60 mmol) of DMAP in 35 ml of absolute toluene and εtirred for 2 hourε at room temperature (25°C) . The reaction mixture is concentrated by evaporation to about 5 ml and filtered on a short εilica gel column (rewaεhed with 30 ml of hexane/ethyl acetate) . Removal of the εolvent and εubsequent column chromatography (hexane/ethyl acetate 4:1) yield 18 mg (34%) of macrolactone 22 as a colorless oil.

Compound 22 (R³ = TBS, R⁴ = Troc)

'H NMR (600 MHZ, CDC1₃) <5 = 6.99 (s, IH) , 6.56 (s, IH) , 5.21-5.14 (m, 2H) , 4.87 (d, J = 12.0 Hz, IH) , 4.75 (d, J = 12 Hz, IH) , 4.05 (d, J = 9.8 Hz, IH) , 3.30 (dq, J = 10.2, 6.3 Hz, IH) , 2.82 (dd, J = 10.3, 4.0 Hz , IH) , 2.79 (dd, J = 16.5, 1.5 Hz , IH) , 2.71 (s, 3H) , 2.64 (dd, J = 16.5, 10.0 Hz, IH) , 2.25-2.21 (m,

IH) , 2.11 (s, 3H) , 1.93-1.64 (m, 4H) , 1.55-1.42 (m, 2H) , 1.32-

1.24 (m, IH) , 1.28 (ε, 3H) , 1.21 (ε, 3H) , 1.19 (s, 3H) , 1.16-1.08

(m, IH) , 1.12 (d, J = 6.7 Hz, 3H) , 1.03 (d, J = 6.7 Hz, 3H) , 0.88

(S, 9H) , 0.16 (s, 3H) , -0.03 (ε, 3H) .

Protection Removal at 3-0

Example: R³ = TBS, R⁴ = Troc

IS mg (23 μmol) of macrolactone 22 iε diεεolved in 0.5 ml of THF and mixed with 2.5 ml of buffered HF-Py (εee above) and εtirred for 72 hourε at room temperature, and then it iε added carefully to 35 ml of a εaturated NAHC0-. εolution and extracted with ether. Removal of the εolvent (repeated εpinning-in with toluene) and εubεequent column chromatography (hexane/ethyl acetate 2:1 — 1:1) yield 5 mg (32%) of deεired product 23 aε a colorless oil. At the same time, 6 mg of starting material iε recovered. Compound 23 (R⁴ = Troc)

¹H NMR (250 MHz, CDCl₃) : δ = 6.99 (ε, IH) , 6.63 (ε, IH) , 5.52 (t, J = 4.5 Hz, IH) , 5.18 (dd, J = 9.0, 2.0 Hz, IH) , 4.84 (d, J = 12 Hz, IH) , 4.78 (d, J = 12 Hz, IH) , 4.15-4.05 (m, IH) , 3.79-3.70 (m, 2H) , 3.64-3.52 (m, IH) , 2.87 (t, J = 6.0 Hz, IH) , 2.72 (ε, 3H) , 2.59 (dd, J = 14.0, 10.0 Hz, IH) , 2.48 (dd, J = 14.0, 4.0 Hz, IH) , 2.35 (t, J = 7.5 Hz, 2H) , 2.12 (ε, 3H) , 2.00 (t, J = 5.5 H∑, 2H) , 1.89-1.81 (m, 2H) , 1.75-1.25 ( , 3H) , 1.39 (ε, 3H) , 1.28 (ε, 3H) , 1.15 (d, J = 7.0 Hz, 3H) , 1.09 (ε, 3H) , 0.9S (d, J = 7.0 Hz, 3H) .

7-O-Protection Removal -* Epothilone B

Example: R⁴ = Troc

2.7 mg (4.4 μmol) of 23 iε diεεolved in 1.5 ml of ethanol, mixed with 25 mg of NH₄C1 and 25 mg of zinc (powder) and refluxed for 30 minuteε. After cooling to room temperature, it iε filtered on Celite, waεhed with ethyl acetate, and the εolvent iε removed in Rotavapor. Subεequent column chromatography (hexane/ethyl acetate 1:1) yieldε 1.4 mg (about 90%) of 24 (epothilone B) . Compound 24 (Epothilone B)

¹H NMR (600 MHz, CDC1₃) δ = 6.97 (ε, IH) , 6.60 (ε, IH) , 5.41 (d, J = 7.8, 2.4 Hz, IH) , 4.27-4.18 (m, 2H) , 3.77 (ε, IH) , 3.30 ( , IH) , 2.81 (dd, J = 7.5, 4.5 Hz, IH) , 2.70 (ε, 3H) , 2.65 (sbr, IH) , 2.54 (dd, J = 14.0, 10.2 Hz, IH) , 2.37 (dd, J = 14.0, 3.0 Hz, IH) , 2.13-2.05 (m, IH) , 2.09 (ε, 3H) , 1.92 (ddd, J = 15.3, 7.7, 7.7 Hz, IH) , 1.78-1.68 (m, 2H) , 1.55-1.46 (m, 2H) , 1.45-1.33 (m, 2H) , 1.37 (ε, 3H) , 1.30-1.22 (m, IH) , 1.28 (s, 3H) , 1.25 (ε, 3H) , 1.17 (d, J = 6.7 Hz, 3H) , 1.08 (s, 3H) , 1.01 (d, J = 7.0 Hz, 3H) .

NMR data are identical to the data of K. C. Nicolaou and A. Mantoulidis (Tet. Lett. 39 (1998) 8633-8636). HPLC analysiε with a comparison sample of A. Mantoulidis shows identical material.

Preparative Methods:

All reactions of organometallic reagentε and all reactionε in abεolute solvents are performed in an air-free and moisture- free environment. The glass equipment that is used iε heated εeveral times in a vacuum (about 0.01 mbar) before the beginning of the test and aerated with dry argon of the Linde Company. Unless otherwise indicated, all reaction batches are εtirred magnetically.

Methylene chloride is predried on a baεic aluminum oxide column of activity εtage I (Woel ) and made absolute on calcium hydride. After predrying on a baεic aluminum oxide column over an 8:1 sodiu /potaεεium alloy, diethyl ether iε refluxed until εtable blue coloring of the benzophenone indicator iε achieved, and it is freεhly distilled off before use. The tetrahydrofuran (THF) is predried over KOH, filtered on a column that iε coated with basic aluminum oxide and then distilled on potassium with triphenylmethane as an indicator.

After predrying over calcium chloride just like hexane (Hex) before use for column chromatography in a rotary evaporator, the ethyl acetate (EE) iε distilled off.

Chromatographic Process:

All reactionε are monitored by thin-layer chromatography (TLC) on silica gel-60-aluminum foils with UV-indicator F₂₅₄ of the Merck Company. Aε a mobile εolvent, in moεt cases εolvent mixtures that conεiεt of hexane (Hex) and ethyl acetate (EE) are used. For viεualization of non-UV-active εubεtanceε, in moεt cases aniεaldehyde/glacial acetic acid/εulfuric acid (1:100:1) haε been taken aε a εtandard dip reagent.

The preparative column chromatography iε performed on εilica gel-60 of the Merck Company (0.04-0.063 mm, 230-400 meεh) , whereby εolvent mixtureε that conεiεt of hexane (Hex) and ethyl acetate (EE) or diiεopropyl ether are uεed aε eluantε.

On an analytical εcale aε well aε on a preparative εcale, the high-preεsure liquid chromatographic εeparationε (HPLC) are performed on modular εyεtemε of the Knauer Company (pump 64 , UV and RI detectorε, columns and recorderε) , Waterε/Millipore Company (injection εyεtem U6K9) and Milton-Roy (integrator CI- 10) . For the analytical HPLC, in moεt caεes a Knauer column (4-250 mm) with 5 μm of nucleoεil iε uεed, and for the preparative HPLC, a column (16^*250 mm, 32^*250 mm or 64-300 mm) with 7 μm or 5 μm nuceloεil 50 iε used.

Dye Reagents

Dye Reagent I (F I) : In the case of moεt compoundε that can be reduced, 1 g of cerium (IV) εulfate in 10 ml of concentrated εulfuric acid and 90 ml of water yield an intenεive blue color reaction during drying.

Dye reagent II (F II) : A 10% ethanolic εolution of molbydatophosphoric acid represents another dip reagent for detecting unsaturated and reducible compounds. In contrast to dye reagent I, the molybdate dye reagentε, eεpecially pertaining to εeveral functionalitieε, εhowε a broader color εpectrum in the case of virtually identical reliability. Dye reagent III (F III) : 1 ml of aniεaldehyde in 100 ml of ethanol and 2 ml of concentrated εulfuric acid repreεents an extremely sensitive dye reagent that in addition alεo εhowε probably the broadeεt color εpectrum.

Dye reagent IV (F IV) : Like the aniεaldehyde reagent, 1 g of vanillin in 100 ml ethanol and 2 ml of concentrated εulfuric acid iε a very εenεitive dye reagent with a broad color εpectrum.

Dye reagent V (F V) : l g of 2 , 4-dinitrophenylhydrazine in 25 ml of ethanol, 8 ml of water and 5 ml of concentrated εulfuric acid represent an excellent dip reagent that reεponds selectively to aldehydes even without being heated and that responds somewhat more εlowly to ketoneε.

Dye reagent VI (F VI) : A 0.5% aqueouε εolution of potaεsium permanganate indicateε groups that can be oxidized by decolorization, whereby unsaturated, non-aromatic structural units react spontaneously without heating.

Spectroscopic Process and General Analysis: NMR-Spectroscopy

The ¹H-NMR spectra are recorded aε an internal standard with a DRX 250 DRX 400 εpectrometer of the Bruker Company with the εubεtanceε aε a εolution in deuterated εolventε and tetramethylεilane. The evaluation of the spectra is carried out according to rules of the first order. If a signal multiplicity that occurs cannot be explained in this way, the indication of the observed line set iε done. To determine the εtereochemiεtry, the NOE-εpectroεcopy (Nuclear Overhauεer Effect) iε uεed. To characterize the signalε, the following abbreviations are used: ε (εinglet) , d (doublet) , dd (double doublet) , ddd (6-line εyεtem with two identical coupling conεtantε or an 8-line εystem in three different coupling constants) , t (triplet) , q (quartet) , quint (quintet) , εext (εextet) , sept (septet) , m (multiplet) , mc

(centered multiplet) , br (broad) , hv (semi-maεked εignal) and v

( aεked εignal) .

The ¹³C NMR εpectra are meaεured aε an internal εtandard with an AC 250 of the Bruker Company with a CDC1₃ εignal at 77.0 ppm, whereby the proton reεonances are wideband-coupled.

IR-spectroscopy

The infrared εpectra are recorded with deviceε of the Perkin-El er Company (model 257 or 580 B) and Nicolet Company (FTIR-interfero eter εyεtem 55XC) . The oils are measured aε films between potaεεium bromide diεks. The bands are indicated according to decreasing wave number (cm^"1). For characterization, the following deεignationε are εelected: vε (very εtrong) , ε (εtrong) , m (medium) , w (weak) .

Abbreviations: abs.: absolute, Ar: aryl/aromatic compound, Cld.: calculated, Brine: cold, saturated common εalt εolution, nBuLi: nbutyllithium, c: concentration, COSY: correlated εpectroscopy (correlated εpectroεcopy) , CSA: ca pherεulfonic acid, TLC: thin- layer chromatography, DCM: dichloromethane, DDQ: dichloro- dicyano-quinone, d.e.: diaεtereomeric excess , DIBAH: diiεobutyl- aluminum hydride, DIPA: diiεopropylamine, DMAP: dimethylaminopyridine, DMF: N,N' -dimethylformamide, DMS : dimethyl sulfide, DMSO: dimethyl εulfoxide, ds: diaεtereoselection, EA: elementary analyεiε, e.e.: enantiomeric exceεε, EE: ethyl acetate, El: electron impact ionization, eq: equivalent (ε) , eV: electron volt, FG: functional group, FI: field ionization, gef.: found, ges.: saturated, h: hour(ε), Hex: n-hexane, HMDS: hexamethyldiεilazide, HPLC: high-preεεure liquid chromatography, Hunig Base: N-ethyl-diisopropylamine, HRMS : high resolution mass εpectrometry, HV: high vacuum, iPrOH: 2-propanol, IR: infrared εpectrometry/infrared εpectrum, J: coupling constant, LDA: lithium diiεopropylamine, Lsg. : εolution, Lsm. : solvent, MC: methylene chloride, Me: methyl, MeLi: methyllithium, min.: minute(s), MS: maεε εpectrometry/maεε εpectra, NMR: nuclear magnetic resonance, NOE: Nuclear Overhauεer Effect, PCC: pyridinium chlorochromate, PG: protective group, Ph: phenyl, ppm: partε per million, Rkt. : reaction, rt. : retention time, RT: room temperature (20-30°C) , Std. : hour(ε), TBAF: tetra-n-butylammonium fluoride, TBDPS: tert-butyldiphenyl- εilyl chloride, TBDPSCl: tert-butyldiphenyl-εilyl chloride, TBS: tert-butyldimethyl-εilyl chloride, TBSCI: tert-butyldi ethyl- εilyl chloride, TBSTriflate: tert-butyldimethyl-silyl-triflate , TEA: triethylamine, tert/t: tertiary, TFA: trifluoroethanoic acid, TFAA: trifluoroethanoic acid anhydride, TFMS: trifluoromethaneεulfonic acid, THF: tetrahydrofuran, TMS: trimethylεilyl-, u: g-mol^'1.

Claims

WHAT IS CLAIMED IS:

1. In a process for the production of epothilone compounds, the improvement comprising preparing εaid compounds by cyclization of a compound produced from an intermediate of formula II

OPG wherein PG iε a protecting group.

2. The process according to claim 1, wherein PG is a TBS or TES group.

3. The process according to claim 1, wherein the compound of formula II contains a TBS group as PG, which group iε changed to a TES group during the process.

4. The process according to claim 1, wherein said cyclization reaction is of a compound of the formula 21