WO1995008553A1

WO1995008553A1 - Synthesis of selectin ligands

Info

Publication number: WO1995008553A1
Application number: PCT/US1994/010790
Authority: WO
Inventors: K. C. Nicolaou; Nicholas J. Bockovich; Daniel R. Carcanague
Original assignee: The Scripps Research Institute
Priority date: 1993-09-22
Filing date: 1994-09-22
Publication date: 1995-03-30
Also published as: AU7878294A

Abstract

The total synthesis of the naturally occurring sulfated Le?x and Lea¿ tetrasaccharides, trisaccharide analogs of sulfated Le?x and Lea¿, and multivalent Lex selectin ligands are described.

Description

SYNTHESIS OF SELECTIN LIGANDS

Specification Field of Invention:

The invention relates to the total syntheses of the naturally occurring sulfated Le and Le" tetrasaccharides and related compounds. More particularly, the invention relates to compounds which are truncated analogs of sulfated Le^x and Le", to key intermediates in the synthesis of sulfated Le^x and Le" tetrasaccharides and related compounds and to multivalent Le^x selectin ligands.

Background of the Invention:

Sialyl Lex-type molecules serve as ligands to E-selectin and contribute to the recruitment of leukocytes to inflammation sites. Several investigators contributed to the initial identification of sialyl Lex-type molecules as ligands to E-selectin, e.g., M. Bevilacqua et al. (Science 1989, 243, 1160), J. Lowe et al. (Cell 1990, 63, 475), M. Phillips et al. (Science 1990, 250, 1130), G. alz et al. (Science 1990, 250, 1132) and M. Tiemeyer et al. (Proc. Nat/. Acad. Sci., USA 1991, 88, 1138). The chemistry and biology of selectins with respect to the recruitment of leukocytes to inflammation sites via vascular adhesion and rolling have been elegantly characterized by L. Lasky (Nature 1991, 349, 196 and Science 1992, 258, 964), by S. Borman (Chem . & Eng . News 1992, 70 (49), 25) and by J. Travis (Science 1993, 260, 906).

More recently, C.-T. Yuen et al. (Biochem. 1992, 31 , 9126) disclosed the isolation of a mixture of two sulfated tetrasaccharides from an ovarian cystadenoma glycoprotein, i.e., sulfated Le^x and sulfated Le". These sulfated compounds exhibited E-selectin binding properties comparable to those of the sialylated compound, e.g. sialyl Lex. Due to the importance of these ligands to adhesion processes and their extreme scarcity, their synthesis has been deemed important and has received considerable attention. Methods for the synthesis of sialyl Le compounds have been disclosed by several workers, e.g., M. Palcic et al. (Carbohydr. Res. 1989, 190, 1) , D. Dumas et al. (Bioorg. Med. Chem. Lett. 1991, 1 , 425), A. Kameya a et al. (Carbohydr. Res. 1991, 209, Cl) , K.C. Nicolaou et al. (J. Am. Chem. Soc. 1992, 1 14, 3126), S. Danishefsky (J. Am. Chem. Soc. 1992, 1 14, 8329 and J. Am. Chem. Soc . 1992, 1 14, 8331), and Y. Ichikawa (J. Am. Chem. Soc . 1992, 1 14, 9283). However, sulfated forms of Le^x and of Le* reported by C.-T. Yuen (supra) have not been disclosed. Given the biological importance of these sulfated molecules and their relative scarcity, there was a great need for a synthetic method for producing such compounds and their analogs.

Summary:

The invention relates to the synthesis of sulfated Le^x-type tetrasaccharides. More particularly, the invention is the total syntheses of sulfated Le^x 1 and of sulfated Le* 2 and related

compounds. Compounds 1 and 2 are tetrameric carbohydrates having four rings, viz. ring A, ring B, ring C, and ring D. The invention includes the synthesis of truncated analogs of sulfated Le^x 1 and sulfated Le" 2 such as compounds 3 and 4, indicated below. Compound 3 is a truncated version of Le^x 1 and compound 4 is a truncated version of Le" 2. The truncated compounds 3 and 4 lack ring A. The invention also includes various key intermediates employed in the syntheses of sulfated Le^x 1 and of sulfated Le* 2, i.e., compounds 5 - 10 and includes multivalent Le^x selectin legands. Compounds 1 - 4 and 5 - 10 are illustrated in Fig. 1.

Synthesis of Sulfated Le^x-type Tetrasaccharide 1:

The synthesis of the sulfated Le^x-type tetrasaccharide 1 is summarized in Scheme I. The glycosyl donor 10 is coupled with the glycosyl acceptor 5 under standard Mukaiyama conditions, i.e., AgC10₄-SnCl₂ (T. Mukaiyama et al., Chem. Lett. 1981, 431) . The resultant glycoside 11 is stereoselectively β-linked and is produced with a 90% yield. The precise chemical mechanism ccounting for the stereoselectivity of this glycoside bond ormation is unknown but presumed to involve the participation of the neighboring group. Treatment of glycoside 11 with MeNHNH₂ in refluxing ethanol resulted in removal of both the acetate and the phthalimide groups leads to the corresponding amino alcohol.

[from D-galactose]

5 Scheme I ^a

e2

[from D-glucosamine]

10

i.e., compound 12. Compound 12 may then be acetylated to give the amide 13 with an overall yield of 80%. Desilylation of amide 13 using fluoride ion leads to hydroxy compound 14 with a yield of 95%. Hydroxyl compound 14 may then be coupled with the galactosyl fluoride 6 to furnish the trisaccharide 15 as a single stereoisomer with a yield of 75%. (See Mukaiyama, supra) The synthesis of galactosyl fluoride 6 is shown in Scheme VI. Selective removal of the allyl protecting group from trisaccharide 15, i.e., H₂Ru(PPh₃)₄ followed by acid hydrolysis, gives the hydroxy compound 16 with a yield of 81%. The hydroxyl compound 16 may then be coupled with the fucosyl fluoride derivative 7 (AgC104-SnCl2) by the method of K.C. Nicolaou et al., (J. Am. Chem. Soc. 1990, 112, 3693) to give, stereoselectively, tetrasaccharide 17 with the desired α-fucose anomeric linkage with a yield of 85%. Reaction of tetrasaccharide 17 with thiourea leads to selective removal of the chloroacetyl group to afford alcohol 18 with a yield of 81%. In turn, alcohol 18 may be converted to the sulfated compound 19, in 95% yield, by exposure to S0₃-NMe₃ complex in anhydrous pyridine. Finally, deacetylation of compound 19 followed by hydrogenolysis gives the targeted sulfated Le^x tetrasaccharide 1 in 80% overall yield. Synthesis of Truncated Sulfated Le^x-type Trisaccharide 3:

The synthesis of the sulfated derivative 3 lacking the galactose unit at the reducing end may be accomplished as depicted in Scheme II using the carbohydrate units 6, 7, and 9 and similar chemistry as described above. Synthesis of carbohydrate unit 9 is described by K.C. Nicolaou et al. (J. Chem. Soc, Chem. Commun . 1991, 870). It is interesting to note that an earlier synthesis of compound 3 by E. Chandrasekaran et al. (J. Biol. Chem . 1992, 267, 23806) was reported prior to the actual elucidation of the natural products, i.e., the sulfated Le^x (1) and sulfated Le* (2).

[from D-galactose]

6

[from L-fucose]

7

[from D-glucosamine]

Synthesis of sulfated Le*-tvpe compounds 2 and 4 Employing the strategy outlined above for synthesizing sulfated Le^x can not be employed for synthesizing sulfated Le*. An attempt to do so led to unexpected glycosidation problems. Accordingly, we developed a different strategy for the synthesis of the sulfated Le*-type compounds 2 and 4. Scheme III summarizes the successful routes to compounds 2 and 4. Thus, coupling of carbohydrate units 6 and 8 under Mukaiyama-Suzuki conditions, i.e. Cp₂HfCl₂-AgOTf, (T. Matsumoto et al. ( Tetrahedron Lett. 1988, 29, 3567) in the presence of 2,6-di-t-butyl-4- methylpyridine leads, stereoselectively, to the /3-glycoside 25 in Sc i θllia

b [^~25 : R1 - R2 - benzylidene 26:Rl-H,F^* -CH2Ph

I — 29 : R1 - Ac; R² - MCA; R3, R4 - Phth

⁹^ : 30:R¹-Ac;R *-H;R3,R4-Phth ^h ^==r 31 : R¹ - Ac; R² - SQ3H; R3, R4 - Phth

I L*. 32:R1 -H;R2-SQ3H;R3 R4_H,Ac-i J2

The precise chemical mechanism accounting for the stereoselectivity of this glycoside bond formation is unknown but presumed to involve the participation of the neighboring group. Regioselective opening of the benzylidene ring by treatment with NaCNBH₃-HCl gives the secondary alcohol 26 in 76% yield. Coupling of secondary alcohol 26 with fucosyl fluoride 7, as disclosed by K.C. Nicolaou et al., (J. Am. Chem. Soc. 1990, 1 12, 3693), leads to the trisaccharide 27 with a yield of 95% with respect to the α-anomer. Trisaccharide 27 may be converted via a DAST-NBS reaction, as disclosed by K.C. Nicolaou et al. (J. Am. Chem. Soc . 1984, 106, 4189) to the glycosyl fluoride 28 in 80% yield. Fluoride 28 can serve as a common precursor to both sulfated Le* 2 and the truncated version of sulfated Le* 4.

The synthesis of the tetrasaccharide 2 is achieved using a seguence involving the coupling of compound 28 with the galactose derivative 5 (Cp₂HfCl₂-Ag0Tf) . This leads, stereoselectively, to compound 29 with a yield of 58%. The chloroacetate moiety can be removed from compound 29 and the sulfate group can be attached in its place (S0₃-NMe₃), furnishing compound 31 via compound 30 with an overall yield of 40%. The phthalimide and acetate groups are both removed from compound 31 by treatment with NH₂NH₂-H₂0 at 100°C. This may then be followed by acetylation of the generated amino group to give the amide 32 in 73% overall yield. Final deprotection to generate the naturally occurring compound 2 is then achieved by hydrogenolysis with a 95% yield.

Synthesis of the trisaccharide 4 may proceed by glycosylating benzyl alcohol with fluoride 28. This leads to compound 33 with a yield of 95%. Compound 33 is then converted to trisaccharide 4 as described above for tetrasaccharide 2 (Scheme III) .

The above syntheses render the natural sulfo- oligosaccharides 1 and 2, as well as their simpler Le^x and Le* sulfate analogs 3 and 4. The products of these syntheses are yielded in pure form suitable for extensive biological investigations. Further studies envisioned in this field may expand the library of biological tools and provide leads for therapeutic agents in the area of inflammation and related conditions.

Brief Description of the Drawings:

Fig.l illustrates sulfated Lewis^x (1,3) and Lewis* (2,4) target molecules and key intermediates (5 - 10) for their chemical synthesis.

Detailed Description:

General Techniques: All reaction were carried out under a dry argon atmosphere using freshly distilled solvents unless otherwise noted. Tetrahydrofuran (THF) was distilled from sodium and benzophenone. Benzene, methylene chloride and toluene were distilled from calcium hydride. All other anhydrous solvents were purchased from Aldrich Chemical Company Inc. Amine bases were dried and stored over potassium hydroxide. Glassware was either oven dried (120°C) or flame dried (0.05 torr) prior to use. Where necessary, compounds were dried by azeotropic removal of water with benzene or toluene under reduced pressure. Reactions were monitored by thin layer chromatography (TLC) on E. Merck silica gel plates (0.25 mm) and visualized using uv light (254 nm) and/or heating with p-anisaldehyde solution (340 mL ethanol, 9.2 mL p-anisaldehyde, 12.5 mL sulfuric acid and 3.75 mL acetic acid). Reaction temperatures were measured externally unless otherwise noted. Solvents used for work-up, chromatography, and recrystallizations were reagent grade from either Fisher Scientific or E. Merck. Reactions were worked- up by washing with saturated agueous solutions of the salts indicated. Flash chromatography was performed on E. Merck silica gel (60, particle size 0.040-0.063 mm). Yields refer to chromatographically and spectroscopically (^JH NMR) pure materials. NMR spectra were recorded on Bruker AMX-500 MHz spectrometer at ambient temperature. Chemical shifts are reported relative to the residual solvent peak. Multiplicities are designated as singlet (s) , doublet (d) , triplet (t) , pseudo triplet (PT) , quartet (q) , multiplet (m) , broad (b) , apparent (app) or obstructed (obs) . IR samples were prepared by evaporation of a solution of the compound in CHC1₃ or CDC1₃ onto a NaCl plate under a stream of argon.1 IR spectra were recorded on a Perkin elmer 1600 series FT-IR spectrophotometer. Optical rotations were measured using a Perkin Elmer 241 polarimeter. High resolution mass spectra (HRMS) were recorded on a VG ZAB-ZSE mass spectrometer under Fast Atom Bombardment (FAB) conditions. Melting points were obtained with a Thomas Hoover Unimelt apparatus and are uncorrected. Microanalyses were performed at the Scripps ?Research Institute.

Abbreviations used herein are as follows: TES = -SiEt₃, TBS = -Si^lBuMe₂, THP = tetrahydropyranyl-, Ts = p-MePhS0₂, DMAP = p -(dimethylamino)pyridine, M.S. = molecular sieves, pyr = pyridine, MEM = methoxyethoxymethyl-, Ms = -S0₂Me, Tf = -S0₂CF₃, PMB = p- methoxybenzyl-, TMS = -SiMe₃.

Sulfated Le^x tetrasaccharide (compound 1) : A method for synthesizing sulfated Le^x tetrasaccharide, i.e. compound 1, is illustrated in Scheme I (supra) . The reaction conditions for each step of Scheme I are provided as follows:

Step "A^M: Compound 10 was combined with 2.0 equivalents of compound 5 with 3.0 equivalents of AgC10₄ and 3.0 equivalents of SnCl₂ with 4A molecular sieves in CH₂C1₂ and allowed to react for 4 hours starting at 0°C and increasing to 25°C to yield compound 11 with a 90% yield.

Step "B": Compound 11 was then combined with a (1:1) mixture of hydrazine-EtOH at 95°C for 48 hours to produce compound 12.

Step "C": Compound 12 was then combined with an excess of Ac₂0 and with an excess of Et₃N, using p- (dimethylamino)pyridine (DMAP) as a catalyst, in CH₂C1₂ at 25°C for 4 hours to produce compound 13 with an overall yield of 80% for the two steps.

Step "D": Compound 13 was then combined with 2.0 equivalents of Bu₄NF in THF at 25°C for 1 hour to produce compound 14 with a yield of 95%.

Step "E": Compound 14 was then combined with 2.0 equivalents of compound 6, 3.0 equivalents of AgC10₄ and 3.0 equivalents of SNC1₂ with 4A molecular sieves in CH₂C1₂ and allowed to react for 5 hours starting at 0°C and increasing to 25°C to yield compound 15 with a 75% yield.

Step "F": Compound 15 was then combined with a catalytic amount of H₂Ru(PPh₃)₄ in EtOH at 95°C for 4 hours; the product was then combined with a catalytic amount of p-TsOH in MeOH at 25°C for 1 hour to produce compound 16 with a 81% yield.

Step "G": Compound 16 was then combined with 2.0 equivalents of compound 7, 3.0 equivalents of AgC10₄ and 3.0 equivalents of SNC1₂ with 4A molecular sieves in CH₂C1₂ and allowed to react for 4 hours starting at 0°C and increasing to 25°C to yield compound 17 with a 85% yield.

Step "H": Compound 17 was then combined with 5.0 equivalents of thiourea, 2.0 equivalents of 2,6- lutidine in EtOH at 65°C for 5 hours to produce compound 18 with an 81% yield.

Step "I": Compound 18 was then combined with 20 equivalents of S0₃-NMe₃ in pyridine at 25°C for 24 hours to produce compound 19 with a 95% yield. Step "J": Compound 19 was then combined with 2.0 equivalents of NaOMe in MeOH at 45°C for 5 hours; and then deprotected with H₂ using Pd(0H)₂ in a 2:1 mixture of MeOH-H₂0 for 48 hours to produce compound 1 with a yield of 80%.

Truncated sulfated Le^x (trisaccharide 3) : A method for synthesizing truncated sulfated Le^x, i.e., the trisaccharide 3, is illustrated in Scheme II (supra). The reaction conditions for each step of Scheme II are provided as follows:

Step "A": Compound 5 was combined with 2.0 equivalents of compound 6 with 3.0 equivalents of AgC10₄ and 3.0 equivalents of SnCl₂ with 4A molecular sieves in CH₂C1₂ and allowed to react for 4 hours starting at 0°C and increasing to 25°C to yield compound 20 with a 81% yield. Step "B": Compound 20 was then combined with a catalytic amount of H₂Ru(PPh₃)₄ in EtOH at 80°C for 1 hour; the product was then combined with a catalytic amount of p-TsOH in a 4:1 mixture of MeOH-CH₂Cl₂ at 25°C for 2 hours to produce compound 21 with a 82% yield. Step "C": Compound 20 was combined with 2.0 equivalents of compound 7 with 3.0 equivalents of AgC10₄ and 3.0 equivalents of SnCl₂ with 4A molecular sieves in a 3:1 mixture of Et₂0-THF and allowed to react for 3 hours starting at -15°C and increasing to 0°C to yield compound 22 with a 85% yield.

Step "D": Compound 22 was then combined with 5.0 equivalents of thiourea, 2.0 equivalents of 2,6- lutidine in a 1:1 mixture of EtOH-CH₂Cl₂ at 65°C for 5 hours to produce compound 23 with a 90% yield. Step "E": Compound 23 was then combined with 20 equivalents of S0₃-NMe₃ in pyridine at 25°C for 24 hours to produce compound 24 with a 86% yield.

Step ^MF": Compound 24 was then combined with 2.0 equivalents of NaOMe in MeOH at 25°C for 4 hours; and then deprotected with H₂ using Pd(OH)₂ in MeOH for 7 days to produce compound 3 with a yield of 74%. Sulfated Le" (tetrasaccharide 2 and trisaccharide 4) :

A method for synthesizing sulfated Le* tetrasaccharide 2 and the trisaccharide 4 is illustrated in Scheme III (supra) . The reaction conditions for each step of Scheme III are provided as follows:

Step "A": Compound 8 was combined with 4.0 equivalents of compound 6, 5.0 equivalents of AgOTf (Tf=-S0₂CF₃) , 5.0 equivalents of Cp₂HfCl₂, and 1.0 equivalent of 2,6-di-t-butyl-4-methylpyridine with 4A molecular sieves in CH₂C1₂ for 6 hours starting at 0°C and ending at 25°C to produce compound 25 with a yield of 63%.

Step "B": Compound 25 was then combined with 10.0 equivalents of NaCNBH₃ and excess ethereal HC1 with 3A molecular sieves in THF at 0°C for 30 minutes to produce compound 26 with a yield of 76%.

Step "C": Compound 26 was then combined with 2.0 equivalents of compound 7 with 4.0 equivalents of AgC10₄ and 4.0 equivalents of SnCl₂ with 4A molecular sieves in a 5:1 mixture of Et₂0-THF and allowed to react for 1 hour starting at -15°C and increasing to 0°C to yield compound 27 with a 95% yield.

Step "D": Compound 27 was then combined with 3.0 equivalents of DAST and 1.25 equivalents of NBS in CH₂C1₂ for 2 hour starting at -78°C and increasing to -20°C to yield compound 28 with a 80% yield.

Step ^,*E": Compound 28 was then combined with 8.0 equivalents of benzyl alcohol, 5.0 equivalents of AgOTf (Tf=-S0₂CF₃) , and 5 equivalents of Cp₂HfCl₂ with 4A molecular sieves in CH₂C1₂ for 18 hours starting at 0°C and ending at 25°C to produce compound 33 with a yield of 95%.

Step "F": Alternatively, compound 28 was then combined with 3.0 equivalents of compound 5, 3.0 equivalents of AgOTf (Tf=-S0₂CF₃) , and 3.0 equivalents of Cp₂HfCl₂ with 4A molecular sieves in CH₂C1₂ for 4 hours starting at 0°C and ending at 25°C to produce compound 29 with a yield of 58%.

Step "G": Compound 29 or 33 was then combined with

5.0 equivalents of thiourea, 2.5 equivalents of 2,6- lutidine in a 1:1 mixture of EtOH-CH₂Cl₂ at 65°C for 12 hours to produce compound 30 with a 79% yield or compound 34 with a yield of 89%, respectively.

Step "H": Compound 30 or 34 was then combined with 20 equivalents of S0₃-NMe₃ in pyridine at 25°C for 24 hours to produce compound 31 with a 50% yield or compound 35 with a 76% yield, respectively.

Step "I": Compound 31 or 35 was then first combined with a 1:1 mixture of hydrazine hydrate-EtOH at 100°C for 3 hours; the product was then combined with an excess of Ac₂0 and with an excess of Et₃N in MEOH at 25°C for 10 minutes to produce compound 32 with an overall yield of 73% for the two steps or to produce compound 36 with an overall yield of 50%, respectively.

Step "J": Compound 32 or 36 was then deprotected with H₂ using Pd(OH)₂ in a 2:1 mixture of MeOH-H₂0 for 48 hours at 25°C to produce compound 2 with a yield of

95% or compound 4 with a yield of 82%, respectively.

Intermediate compounds 8 and 10: A method for synthesizing intermediate compounds 8 and 10 is illustrated in Scheme IV (supra) . The reaction conditions for each step of Scheme IV are provided as follows:

Step "A": Compound 37 was combined with 3.0 equivalents of benzaldehyde dimethyl acetal, with a catalytic amount CSA in THF for 16 hours at 55°C to produce compound 8 with a yield of 85%.

Step "B": Compound 8 was then combined with 2.0 equivalents of NaH, 2.0 equivalents of allyl bromide, and 0.1 equivalent of Bu₄NI in THF for 16 hours starting at 0°C and ending at 50°C to produce compound

38 with a yield of 91%.

Step "C": Alternatively, compound 37 was combined with 10.0 equivalents of NaCNBH₃ and excess ethereal HCl with 3A molecular sieves in THF for 1 hour starting at 0°C and ending at 25°C to produce compound 39.

Step "D": Compound 39 was then combined with 1.5 equivalents of t-butyldimethylsilyl triflate (trifluoromethanesulphonate) , 1.7 equivalents of 2,6- lutidine in CH₂C1₂ for 1.5 hours starting at 0°C and ending at 25°C to produce compound 40 with a 75% yield for two steps. Step "E": Alternatively, compound 39 was then combined with 3.0 equivalents of DAST and 1.2 equivalents of dimethyl(methylthio)sulfonium triflate (trifluoromethanesulphonate) , i.e., DMTST, in CH₂C1₂ for 1 hour at -10°C to produce compound 10 with a 50% yield.

Intermediate compound 5: A method for synthesizing intermediate compound 5 is illustrated in Scheme V (supra) . The reaction conditions for each step of Scheme V are provided as follows:

Step "A": Compound 41 was combined with excess 2,2-dimethoxypropane and a catalytic amount CSA in acetone for 48 hours at 25°C to produce compound 42 with a yield of 56%. Step "B": Compound 42 was then combined with 3.0 equivalents of NaH, 3.0 equivalents of benzyl bromide, and a catalytic amount Bu₄NI in THF for 2 hours starting at 0°C and ending at 50°C to produce compound 43 with a yield of 89%. Step "C": Compound 43 was then combined with 3.0 equivalents of DAST and 1.2 equivalents of NBS in CH₂C1₂ for 4 hour starting at -5°C and increasing to 10°C to yield compound 44 with a 80% yield.

Step "D": Alternatively, compound 41 was then combined with 3.0 equivalents of benzyl alcohol, 3.0 equivalents of AgC10₄, and 1.5 equivalents of Cp₂ZrCl₂ with 4A molecular sieves in benzene for 2 hours to NOT FURNISHED UPON FILING

combined with an excess of HF-pyridine and 1.3 equivalents of NBS in CH₂C1₂ for 6 hour starting at - 78°C and increasing to 0°C to yield compound 6 with a 76% yield.

The chemical intermediates were physically characterized as follows:

11

11: R_f = 0.45 (silica, 95:5, dichloromethane: ether) ; [α]_D ²⁵ +62.2 ° (c = 0.785, chloroform) : IR (film) v_maχ 3029 (w) , 2928 (m) , 2959 (m) , 1775 ( ) , 1746 (m) , 1715 (s), 1457 (m) , 1386 (s), 1223 (s) , 1106 (s) , 1061 (m) cirf ¹; NMR (500 MHz, CDC1₃) d 7.81-7.60 (m, 4 H, aromatic), 7.37-7.07 (m, 18 H, aromatic) , 6.83-6.81 (m, 2 H, aromatic) , 5.55-5.47 (m, 2 H, H-4A, CH=CH₂) , 5.36 (d, J - 7.8 Hz, 1 H, H-1B) , 4.89 (m, 1 H, -CH=CH₂ ) , 4.71 (m, 1 H, -CH=CH₂) , 4.66-4.58 (m, 3 H, OCH₂Ph) , 4.55 (d, J^" = 3.8 Hz, 1 H, H-IA) , 4.48-4.36 (m, 3 H, OCH₂Ph) , 4.18-4.08 (m, 5 H, CHO) , 4.02 (dd, J = 4.3, 7.0 Hz, 1 H, CHO) , 3.85 (d, J = 12.7 Hz, 1 H, OCH₂-CH=CH₂) , 3.78-3.54 (m, 5H, CHO) , 3.49 (dd, J = 3.8, 10.0 Hz, 1 H, H-2A) , 3.43 (dd, J^" = 3.7, 10.1 Hz, 1 H, H-3A) , 3.34 (dd, J = 7.2, 10.2 Hz, 1 H, CHO) , 2.04 (s, 3 H, acetate) , 0.85 (s, 9 H, -SiC(CH₃)₃) , 0.07, 0.04 (s, 3 H each, SiCH₃) ; ¹³C NMR (125 MHz, CDC1₃) 3 170.2, 138.8, 138.2, 138.0, 136.7, 134.4, 133.9, 131.7, 128.5, 128.3, 128.25, 128.20, 128.1, 128.0, 127.8, 127.52, 127.49, 127.34, 127.28, 127.24, 127.20, 127.0, 123.2, 116.4, 99.0, 95.5, 79.7, 76.2, 75.5, 75.1, 73.6, 73.3, 73.2, 72.8, 71.8, 71.5, 69.4, 69.1, 68.8, 68.3, 56.5, 25.9, 20.8, 17.9, -4.0, -4.6; HRMS (LSIMS) Calcd for C₅₉H₆₉N0₁₃SiCs (M+Cs) : 1160.3593, found: 1160.3586.

15

15: Rf = 0.20 (silica, 1:1, ethyl acetate :petroleum ether) ; [α]_D ²⁵ +24.5 o (c = 2.2, chloroform) : IR (film) ^vmax ^{3389 (w)} ' ^{2924 ( )} ' ²⁸⁷° ^(w) ' ^{1748 (s)} ' ^{1667 (m)} ' 1538 (w) , 1371 (s) , 1228 (s) , 1056 (s) cm^-1; ^XH NMR (500

MHz, CDC1₃) δ 7.37-7.26 (m, 20 H, aromatic) , 5.84 (m, 1 H,

-CH=CH₂) , 5.64 (d, J = 9.0 Hz, 1 H, NH) , 5.45 (d, J, 3.0

Hz, 1 H, H-4A) , 5.31 (d, J = 3.2 Hz, 1 H, H-4C) , 5.23 (m,

1 H, -CH=CH₂) , 5.16-5.08 (m, 2 H, -H-2C, -CH=CH₂ ) , 4.95

(dd, J = 3.3, 10.5 Hz, 1 H, H-3C) , 4.86-4.83 (m, 2 H,

OCH₂Ph) , 4.70-4.63 (m, 3 H, OCH₂Ph) , 4.55-4.44 (m, 6 H,

CHO) , 4.19-4.05 ( , 6 H, CHO) , 3.99-3.94 ( , 3 H, CH₂C1,

CHO) , 3.87-3.65 (m, 5 H, CHO) , 3.62 (t, J = 6.6 Hz, 1 H,

CHO) , 3.52-3.39 (m, 3 H, CHO) , 2.13, 2.06, 2.03, 1.99,

1.80 (s, 3 H each, acetyls ¹³C NMR (125 MHz, CDCI₃) 3 170.4, 170.2, 170.1, 169.84, 169.76, 166.5, 138.3, 138.2, 137.9, 136.9, 135.0, 128.7, 128.52, 128.49, 128.43, 128.40, 128.35, 128.29, 128.26, 128.20, 127.07, 127.95, 127.90, 127.84, 127.77, 127.72, 127.69, 127.64, 127.60, 127.56, 127.38, 127.29, 116.3, 101.0, 99.6, 95.6, 78.3, 76.1, 75.0, 74.9, 74.2, 73.6, 73.3, 72.8, 72.4, 71.6, 70.9, 70.4, 69.2, 69.1, 68.8, 68.5, 66.6, 60.8, 53.0, 40.4, 29.6, 23.2, 20.9, 20.7, 20.6; HRMS (LSIMS) Calcd for C₆₁H₇₂N0₂₁ClCs (M+Cs) : 1322.3340, found: 1322.3301.

18: R_f = 0.20 (silica, 1:1, ether :dichloromethane) ;

[ ]_O ^z^ +4.4 ° (c = 1.00, chloroform) : IR (film x 3377 (s) , 2922 (m) , 2866 (m) , 1742 (s) , 1661 (m) , 1365 (m) , 1230 (s) , 1096 (s) , 1049 (s) , 739 (m) cm^-1; ^XH NMR (500 MHz, CDC1₃) δ 7.43-7.19 (m, 35 H, aromatic) , 5.70 (d, J^" = 8.2 Hz, 1 H, NH) , 5.48 (d, J = 2.9 Hz, 1 H, H-4A) , 5.21 (d, J = 3.3 Hz, 1 H, H-4C) , 5.19 (d, J = 3.6 Hz, 1 H, H- 1D) , 5.03 (d, J = 6.2 Hz, 1 H, H-1B) , 4.49 (d, J = 11.8 Hz, 1 H, OCH₂Ph) , 4.87-4.63 (m, 9 H, CHO) , 4.58-4.42 (m, 7 H, CHO) , 4.37 (d, J^" = 12.0 Hz, 1 H, OCH₂Ph) , 4.17-3.97 (m, 7 H, CHO) , 3.89 (dd, J - 2.2, 10.1 Hz, 1 H, CHO) , 3.81- 3.53 (m, 7 H, CHO) , 3.47-3.37 (m, 3 H, CHO) , 2.62 (bs, 1 H, OH) , 2.04, 2.02, 1.99, 1.91, 1.64 (s, 3 H each, acetyls) , 1.13 (d, J^" = 6.4 Hz, 3 H, H-6D) ; ¹³C NMR (125 MHz, CDCI3) δ 170.7, 170.6, 170.4, 170.1, 169.7, 138.9, 138.7, 138.5, 138.3, 138.1, 137.9, 136.9, 128.7, 128.60, 128.56, 128.48, 128.45, 128.42, 128.38, 128.34, 128.29, 128.24, 128.20, 128.16, 128.10, 128.06, 128.04, 127.96, 127.90, 127.86, 127.82, 127.76, 127.73, 127.65, 127.58, 127.53, 127.47, 127.20, 127.16, 127.03 126.93, 100.2,

99.1, 96.7, 95.7, 79.6, 76.0, 75.9, 75.4, 74.6, 74.5,

74.2, 73.8, 73.7, 73.4, 73.1, 72.9, 72.8, 72.6, 72.4, 71.0, 70.7, 70.4, 69.2, 69.0, 68.4, 68.0, 66.1, 60.6, 29.6, 23.1, 20.8, 20.7, 20.64, 20.61, 16.7; HRMS (LSIMS) Calcd for C₈₃H₉₅N0₂₄Cs :M+CS) 1622.5298 found 1622.5304.

21

2 1 : R. 0.25 ( silica 4 : 1 dichloromethane :acetone) ; [α]_D ²⁵ +75.9 ° (c = 1.00, chloroform) : IR (film) v_maχ 3487 (s) , 2927 (s) , 1750 (s) , 1665 (m) , 1536 (m) , 1372 (s), 1223 (s), 1047 (s) , 748 (m) cm^-1; ¹H NMR (500 MHz, CDCI3 ) δ 7.42-7.27 (m, 10 H, aromatic) , 5.65 (bd, 1 H, NH) , 5.33 (d, J = 3.3 Hz, 1 H, H-4C) , 5.19 (dd, J = 8.1, 10.4 Hz, 1 H, H-2C) , 4.97 (d, J = 2.6 Hz, 1 H, H-1B) , 4.93 (dd, J = 3.4, 10.4 Hz, 1 H, H- 3C) , 4.77 (d, J = 12.1 Hz, 1 H, OCH₂Ph) , 4.69 (d, J^" = 11.8 Hz, 1 H, OCH₂Ph) , 4.49-4.44 (m, 2 H, OCH₂Ph) , 4.40 (d, J = 8.1 Hz, 1 H, H-1C), 4.20-4.16 (m, 1 H, CHO) , 4.11 (d, J = 6.5 Hz, 2 H, H-6C), 3.95 (s, 2 H, CH₂C1) , 3.88 (bt, J^" = 6.5 Hz, 1 H, H-5C) , 3.82-3.71 (m, 3 H, CHO) , 3.66 (dd, J = 2.5, 10.8 Hz, 1 H, H-6B) , 3.58 (d, J = 10.8 Hz, 1 H, H-

6B) , 2.14, 2.06, 1.98, 1.97 (s, 3 H each, acetyls! 13 C

NMR (125 MHz, CDC1₃) δ 170.4, 170.2, 170.1, 169.1, 166.5, 137.9, 137.0, 128.54, 128.50, 128.05, 127.97, 127.89, 127.87, 127.82, 101.0, 96.8, 81.1, 73.6, 72.4, 70.9, 70.3, 69.9, 69.7, 68.3, 67.5, 66.6, 61.1, 52.8, 40.2, 23.3, 20.7, 20.6, 20.5; HRMS (LSIMS) Calcd for C₃₆H₄₄N0₁₅ClCs (M+Cs) ; 898.1454, found 898.1456.

22

22: R_f = 0.70 (silica, 1:1, ether : dichloromethane) ; [α]_D ²⁵ +7.2 ° (c = 1.07, chloroform) : IR (film) v_maχ 3031 (s) , 2932 (m) , 1752 (s) , 1628 (m) , 1551 ( ) , 1370 ( ) , 1221 (s), 1048 (s) cm^-1; ^XH NMR (500 MHz, CDC1₃) δ 7.43- 7.12 (m, 25 H, aromatic) , 6.65 (d, J = 7.3 Hz, 1 H, NH) , 5.23 (d, J - 3.4 Hz, 1 H, H-1D) , 5.20 (d, J - 3.3 Hz, 1 H, H-4C) , 5.08 (d, J = 3.6 Hz, 1H, H-1B) , 5.01 (dd, J = 8.1, 10.3 Hz, 1 H, H-2C) , 4.95 (d, J = 11.6 Hz, 1 H, OCH₂Ph) , 4.80 (d, J = 12.1 Hz, 1 H, OCH₂Ph) , 4.78-4.72 (m, 3 H, CHO) , 4.71-4.63 (m, 4 H, OCH₂Ph) , 4.54 (d, J = 8.1 Hz, 1 H, H-1C) , 4.50 (d, J = 12.1 Hz, 1 H, OCH₂Ph) , 4.41 (d, J = 12.1 Hz, 1 H, OCH₂Ph) , 4.27 (bq, J = 6.5 Hz, 1 H, H-5D) , 4.15-4.08 (m, 3 H, CHO) , 4.03-3.97 (m, 3H, CHO) , 3.94-3.91 (m, 3 H, CH₂C1, CHO) , 3.77 (dd, J = 2.8, 11.1 Hz, 1 H, H-6B) , 3.70 (d, J - 1.7 Hz, 1 H, H-4D) , 3.63- 3.60 ( , 1 H, H-5B) , 3.55 (dd, J = 1.6, 11.1 Hz, 1 H, H- 6B) , 3.45 (bt, J = 7.3 Hz, 1 H, CHO) , 1.99, 1.97, 1.96, 1.55 (s, 3 H each, acetyls) , 1.22 (d, J = 6.5 Hz, 3 H, H- 6D) ; ¹³C NMR (125 MHz, CDC1₃) δ 170.4, 170.1, 169.9, 168.9, 166.5, 138.55, 138.52, 137.7, 137.5, 137.4, 128.7, 128.62, 128.58, 128.55, 128.45, 128.41, 128.32, 128.30, 128.26, 128.23, 128.20, 128.16, 128.07, 128.04, 127.98, 127.94, 127.86, 127.82, 127.67, 127.55, 127.43, 127.40,

127.37, 127.28, 127.03, 99.2,

77.2, 77.1, 75.3, 75.0, 746.,

70.9, 70.4, 70.0, 69.1, 67.6,

22.7, 20.8, 20.6, 20.5, 16.8 HRMS (LSIMS) Calcd for

C₆₃H₇₂N0₁₉ClCs (M+Cs) : 1314.3441 found: 1314.3451.

25

25: R_f = 0.35 (silica, 95:5, dichloromethane : ether) ; [α]_D ²⁵ +24.0 ° (c = 1.00, chloroform) : IR (film) v_maχ 3349 (br) , 3026 (m) , 2962 (m) , 1749 (s) , 1715 (s) , 1377 (m) , 1221 (s) , 1101 (s) cm^-1; ¹H NMR (500 MHz, CDC1₃) δ 7.94- 7.77 (m, 4 H, aromatic) , 7.49-7.23 (m, 10 H, aromatic) , 5.58 (s, 1 H, CHPh) , 5.57 (d, J^" = 6.7 Hz, 1 H, H-1B) , 5.18 (d, J^" = 3.4 Hz, 1 H, H-4C) , 5.01 (dd, J = 8.0, 10.4 Hz, 1 H, H-2C) , 4.80 (dd, J = 3.4, 10.4 Hz, 1 H, H-3C) , 4.75 (dd, J = 8.9, 9.7 Hz, 1 H, CHO) , 4.55 (d, J = 8.0 Hz, 1 H, H-1C) , 4.43-4.38 ( , 2 H, CHO) , 4.03 (dd, J = 8.3, 11.1 Hz, 1 H, H-6C), 3.88-3.79 (m, 5 H, CH₂C1, CHO), 3.75-3.72 ( , 1 H, CHO) , 3.48 (m, 1 H, CHO) , 2.08, 1.92, 1.53 (s, 3 H each, acetates) ; ¹³C NMR (125 MHz, CDC1₃) 5 170.5, 169.9, 168.7, 166.5, 136.9, 132.75, 132.72, 131.3, 129.3, 129.0, 128.9, 128.41, 128.38, 128.35, 128.2, 126.02, 125.99, 125.94, 101.5, 100.2, 91.9, 84.2, 80.7, 76.6, 72.7, 72.5, 71.5, 70.5, 70.1, 68.8, 68.5, 67.4, 66.5, 66.3, 60.8, 60.5, 54.2, 40.2, 20.6, 20.5, 20.0; HRMS (LSIMS) Calcd for C₄₁H_4QN0₁₅ClSCs (M+Cs) : 986.0862, found: 986.0868.

27

27 : R_f = 0.40 (silica, 36 :65, ethyl acetate:petroleum ether) ; [α]_D ²⁵ -8.7 ° (c = 2.40, chloroform) : IR (film) v_maχ 3027 (m) , 2930 (m) , 2873 ( ) , 1752 (s) , 1715 (s), 1607 ( ) , 1452 (m) , 1376 (s) , 1219 (s) , 1076 (s) , 740 (s) cm^-1; ¹H NMR (500 MHz, CDC1₃ ) δ 7.90-7.80 (m, 4 H, aromatic) , 7.45-7.11 (m, 25 H, aromatic) , 5.32 (d, J = 10.4 Hz, 1 H, H-l B) , 5.18 (d, J = 3.4 Hz, 1 H, H-4C) , 5.17 (d, J = 3.9 Hz, 1 H, H-1D) , 5.02 (dd, J = 8.2, 10.4 Hz, 1 H, H-2C) , 4.98 (d, J^" = 12.3 Hz, 1 H, OCH₂Ph) , 4.90 (d, J = 11.6 Hz, 1 H, OCH₂Ph) , 4.85-4.68 (m, 7 H, CHO), 4.46 (dd, J^" = 3.4, 10.4 Hz, 1 H, H-3C) , 4.40 (s, 2 H, OCH₂Ph) , 4.35 (t, J = 10.3 Hz, 1 H, CHO) , 4.25-4.17 (m, 3 H, CHO) , 4.02-3.94 (m, 4 H, CHO) , 3.81 (s, 2 H, CH₂C1), 3.75 (bs, 1 H, H-4D) , 3.70-3.62 (m, 2 H, CHO) , 2.00, 1.98, 1.70 (s, 3 H each, acetates) , 1.28 (d, J = 6.5 Hz, 3 H, H-6D) ; ¹³C NMR (125 MHz, CDC1₃ ) δ 169.9, 169.8, 169.2, 166.5, 138.6, 138.2, 138.15, 132.5, 131.6, 128.75, 128.70, 128.6, 128.43, 128.40, 128.36, 128.30, 128.25, 128.22, 128.17, 128.14, 128.11, 127.93, 127.75, 127.59, 127.52, 127.49, 127.46, 127.42, 127.36, 127.29, 127.26, 126.9, 123.7, 100.4, 97.7, 91.8, 83.3, 80.8, 79.9, 79.0, 77.2, 76.4, 76.0, 75.8, 75.4, 74.8, 74.7, 73.7, 73.4, 73.0, 72.9, 72.7, 72.6, 72.1, 70.0, 67.3, 66.6, 66.2, 66.0, 59.5, 55.2, 40.2, 20.5, 20.4, 16.9, 16.7; HRMS (LSIMS) Calcd for C₆₈H₇₀NO₁₉SClCs (M+Cs) : 1404.3006, found: 1404.2911.

29: R_f = 0.33 (silica, 93:7, dichloromethane: ether) ;

[α]_D ²⁵ -7.9 ° (c = 1.00, chloroform) : IR (film) v_maχ 3029 (w) , 2930 (m) , 2872 (m) , 1751 (s) , 1716 (s) , 1454 (m) , 1375 (s) , 1222 (s) , 1072 (s) , 741 (s) cm^-1; ^XH NMR (500 MHz, CDC1₃) δ 7.52-6.90 (m, 39 H, aromatic), 5.30 (d, J = 3.7 Hz, 1H, H-4A) , 5.13 (d, J = 3.9 Hz, 1 H, H-1D) , 5.10 (m, 2 H, H-1B, H-4C) , 4.92 (dd, J = 8.1, 10.4 Hz, 1 H, H- 2C) , 4.90 (d, J = 12.2 Hz, 1 H, OCH₂Ph) , 4.82-4.63 ( , 8 H, CHO) , 4.48-4.34 (m, 8 H, CHO), 4.24 (d, J^" = 7.8 Hz, 1 H, H-IA) , 4.18-4.07 ( , 3 H, CHO) , 3.97-3.91 (m, 2 H, CHO) , 3.87 (d, J = 10.7 Hz, 2 H, H-6A) , 3.73 (s, 2 H, CH₂C1) , 3.67-3.42 (m, 9 H, CHO) , 3.28 (dd, J - 7.8, 9.5 Hz, 1 H, H-2B) , 1.97, 1.91, 1.88, 1.62 (s, 3 H each, acetates) , 1.19 (d, J^" = 6.6 Hz, ? H, H-6D) ; ¹³C NMR (125 MHz, CDCI3) δ 170.1, 170.0, 169.8, 169.1, 166.6, 138.7, 138.6, 138.5, 138.21, 138.17, 137.8, 136.7, 134.4, 128.7, 128.41, 128.38, 128.36, 128.32, 128.28, 128.22, 128.19, 128.14, 128.08, 128.04, 128.00, 127.85, 127.75, 127.72, 127.67, 127.58, 127.54, 127.4, 127.3, 127.2, 127.1, 126.9, 126.8, 123.3, 102.2, 100.2, 98.4, 97.5, 80.8, 78.4, 77.2, 76.1, 75.6, 75.3, 74.8, 74.2, 73.8, 73.6, 72.9, 72.8, 72.7, 72.5, 72.1, 71.0, 70.0, 69.9, 69.0, 67.4, 67.1, 66.1, 66.0, 59.5, 56.6, 40.2, 20.7, 20.5, 20.4, 20.3, 16.9; HRMS (LSIMS) Calcd for ¹³C₁C₉₀H₉₆NO₂₆ClCs (M+Cs) : 1786.4963, found: 1786.5099.

R. 0.45 ( silica 40 : 55 : 5 chloroform:methanol :water) ; [ ]^^2S -39.1 ° (c = 0.85, ethanol) : IR (KBr) v_maχ 3396 (br) , 1653 (s) , 1221 (m) , 1089 (s) , 762 (m) , 619 (m) cm^"1; ¹H NMR (500 MHz, D₂0) δ 5.16 (d, J = 3.7 Hz, 0.5 H, H-lAα) , 5.07 (d, J = 3.9 Hz, 1 H, H-1D) , 4.66 (d, J = 8.5 Hz, 0.5 H, H-lAβ) , 4.52 (d, J = 8.4 Hz, 1 H, H-1C) , 4.50 (d, J = 8.4 Hz, 1 H, H-1B) , 4.26 (dd, J = 3.2, 9.9 Hz, 1 H, H-3C) , 4.21 (d, J = 3.2 Hz, 1 H, H-4C) , 4.15 (d, J = 3.1 Hz, 0.5 H, H-4Aβ) , 4.10 (d, J = 3.3 Hz, 0.5 H, H-4Aα) , 4.05-3.53 ( , 17 H, CHO) , 3.46 (dd, J = 8.3, 9.9 Hz, 1 H, H-2B) , 1.97 (s, 3 H, acetyl) , 1.12 (d, J - 6.6 Hz, 3 H, H-6D) , ¹³C NMR (125 MHz, CD₃OD) δ 168.9, 103.7, 100.1, 93.9, 82.1, 76.2, 76.1, 74.5, 73.5, 72.7, 71.0, 69.7, 68.2, 67.8, 62.7, 62.4, 57.5, 23.2, 16.5; MS (LSIMS) : 770 (M-H) .

2: R_f = 0.21 (silica, 2:2:1, ethyl acetate:2- propanol: water) ; [α]_D ²⁵ -30.9 ° (c = 0.75, methanol) : IR

(film) v_maχ 3382 (br) , 2928 (w) , 1639 ( ) , 1232 (m) , 1069

(s) , 813 (m) , 709 (w) cm^"1; ¹H NMR (500 MHz, D₂0) δ 5.19

(d, J = 3.8 Hz, 0.5 H, H-lAα) , 5.00 (d, J = 3.8 Hz, 1 H,

H-1D) , 4.83 (bq, J = 6.5 Hz, 1 H, H-5D) , 4.69 (d, J = 8.4

Hz, 0.5 H, H-lAβ) , 4.58 (d, J = 7.7 Hz, 1 H, H-1C) , 4.53

(d, J = 7.9 Hz, 1 H, H-1B) , 4.29-4.24 (m, 2 H, H-3C, H-

4C) , 4.18-4.05 (m, 2 H, CHO) , 3.94-3.49 (m, 17 H, CHO) ,

2.02 (s, 3 H, acetyl) , 1.15 (d, J^" = 6.5 Hz, 3 H, H-6D) ;

¹³C NMR (125 MHz, CD₂OD) δ 71.2 104.5, 103.8 99.6, 98.8,

94.3, 84.1, 82.1, 80.7, 78.0, 77.2, 76.2, 73.7, 73.4,

72.8, 71.2, 70.6, 70.0, 69.9, 69.4, 68.0, 67.7, 62.7,

62.6, 62.5, 61.1, 57.5, 23.3, 16.6; HRMS (LSIMS Calcd for C₂₆H₄₄N0₂₃SCs₂(M-H+2Cs) : 1036.0134 found 1036.0134.

3 : R, 0.15 ( silica 60 :35 : 5, chlorof orm:methanol :water ) ; [α]_D ²⁵ -23.2 ° (c = 0.60, ethanol) : IR (film) _γ 3367 (br) , 2937 (m) , 1642 (s),

1549 ( ) , 1425 (w) , 1378 (m) , 1241 (s), 1072 (s), 816 (m) cm^"1; ~H NMR (500 MHz, CD3OD) δ 5.02 (m, 1 H, H-ID) , 4.96

(d, J = 3.3 Hz, 0.5 H, H-lBα) , 4.86 (bq, J = 6.6 Hz, 1 H,

H-5D) , 4.61 (d, J = 8.0 Hz, 0.5 H, H-lBβ) , 4.55 (m, 1 H

H-1C) , 4.23-4.14 (m, 3 H, CHO), 4.05-4.00 (m, 2 H, CHO),

3.97-3.83 (m, 4 H, CHO), 3.80-3.58 (m, 6 H, CHO) , 3.50

(bt, J = 6.0 Hz, 1 H, CHO), 1.97 (s, 3 H, acetyl) , 1.16

(m, 3 H, H-6D) ; ¹³C NMR (125 MHz, CD₃OD) δ 173.9, 103.7,

100.4, 93.0, 82.3, 77.4, 76.3, 76.2, 75.5, 75.1, 74.4,

73.7, 72.8, 71.2, 71.1, 70.0, 69.9, 68.2, 67.6, 62.7,

61.3, 55.8, 22.8, 16.6; HRMS (LSIMS) Calcd for C₂₀H₃₅NO₁₈S

(M+) :609.1575, found 609.1598.

4 : R_f = 0.14 (silica, 60 : 35 : 5 , chlorof orm:methanol :water) ; [α]_D ²⁵ -43.3 ° (c = 0.42, methanol) : IR (film) v_maχ 3450 (br) , 2921 (w) , 1639 (m) , 1224 (m) , 1072 (s) , 1033 (s) cm^"1, ^XH NMR (500 MHz, CD₃OD) δ 5.08 (bs, 0.5 H, H-lBα) , 4.99 (bs, 1 H, H-ID) , 4.68 (d, J = 8.6 Hz, 0.5 H, H-lBβ) , 4.56 (m, 1 H, H-IC) , 4.30-3.50 (m, 16 H, CHO) , 2.01 (s, 3 H, acetyl), 1.15 (d, J = 6.2 Hz, 3 H, H-6D) ; ¹³C NMR (125 MHz, CD₃OD) δ 172.4, 104.7, 99.6, 96.7, 92.9, 82.4, 76.3, 76.1, 74.0, 73.7, 72.9, 71.2, 70.5, 70.1, 68.1, 67.6, 62.8, 61.5, 58.9, 55.7, 22.9, 16.6; HRMS (LSIMS) calcd for C₂₀H₃₄NO₁₈SCs₂ (M- H+2CS) : 873.9605, found 873.9601.

Claims

What is claimed is:

1. Compound 5 having the following structure:

2 . A method for making compound 5 comprising the following steps :

_a I — 41 : Ri = R = H; X = SP

■ — 42 : R- = acetonide;

^•— *- 43 : Ri = acetonide; R = Bn; X = SPh ^c L_*. 44 : R₁ = acetonide; R = Bn; X = F

3. Compound 6 having the following structure:

4. A method for making compound 6 comprising the following steps:

Compound 8 having the following structure :

8

6. A method for making compound 8 comprising the following steps :

7. Compound 10 having the following structure:

tBuMe

[from D-glucosamine]

10

8. A method for making compound 10 comprising the following steps:

Compound 4 having the following structure:

- 36/1 -

10 . A method for making compound 4 compris ing the following steps :

_d r~ 27 : R¹ = Ac; R² = MCA; R³, R⁴ = Phth; X = SPh _e r— 28 : R¹ = Ac; R² = MCA; R³, R⁴ = Phth; X = F r— 33 : R¹ = Ac; R² = MCA; R³, R⁴ = Phth; X = OBn ⁹ 34 : R¹ = Ac; R² = H; R³, R⁴ = Phth; X = OBn

35 : R¹ = Ac; R² = SO₃H; R³, R⁴ = Phth; X = OBn

36 : R = H; R² = S0₃H; R³, R⁴ = H, Ac; X = OBn □ - 37 -

₁₁. _A method for making compound 3 comprising the following steps :

- 38 -

12 . A method for making compound 2 comprising the following steps :

--Z 25 : R¹ = R² = benzylidene

26 : R¹ = H, R² = CH₂Ph

_g Ac; R² = MCA; R³, R⁴ = Phth _h Ac; R - = H; R³, R⁴ = Phth . Ac; R² = S0₃H; R³, R⁴ = Phth j '

H; R² = S0₃H; R³, R⁴ = H, Ac -ξ) - 39 -

13. A method for making compound 1 comprising the following steps:

15 : R = Allyl ^fC 16:R = H