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I? 4 "~51 llllllllllllllllilllllllllllllllllll 3 1293 01051 7765 This is to certify that the dissertation entitled Synthetic Approaches to the Structural Analogs of Lipid A and Total Syntheses of Structurally Non—related Lipid A Antagonists presented by Kyung-Il Kim has been accepted towards fulfillment of the requirements for Ph. D. degreein DChemistry D . Rawle I. Hollings- [ —worth (a I'd/dill; M4” Major professoy Date 3-28-1994 MS U is an Affirmative Action/Equal Opporlunity Institution 0-12771 _ LIBRARY Michigan State University PLACE IN RETURN BOX to rem ove this checkout from your record. TO AVOID FINES9 return on or before date due.= [ DATE DUE DATE DUE DATE DUE I la | i ‘ l “—' J ___l _1 I-i—i: “7| i“ I l MSU Is An Affirmative Action/Equal Opportunity Institution WWma-p. t :- SYNTHETIC APPROACHES TO THE STRUCTURAL ANALOGS OF LIPID A AND TOTAL SYNTHESES OF STRUCTURALLY NON-RELATED LIPID A ANTAGONISTS BY KYUNG-IL KIM A DISSERTATION Submitted to Michigan State University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Chemistry 1994 ABSTRACT SYNTHETIC APPROACHES TO THE STRUCTURAL AN ALOGS OF LH’ID A AND TOTAL SYNTHESES OF STRUCTURALLY NON -RELATED LIPID A ANTAGONISTS By Kyung-ll Kim The lipid A moiety of lipopolysaccharides (LPS ) of gram-negative bacteria (endotoxin ) have tremendous potential as therapeutic agents. They are usually diphosphorylated glucosamine disaccharides which are esterified and amidated with fatty acid chains. Despite their potential, the use of these molecules has been severely limited because preparations from bacterial sources are very variable, ill-defined, heterogeneous and contain contaminants which have deleterious effects. The optimum structure of the active components of these complex mixtures and their modes of action are still to be determined. Biological studies with well defined, pure lipid A species are necessary for obtaining structure activity relationship information. In order to test various theories on structure and how it determines reactivity, we designed two new lipid A analogs, WI and TM2, in which phosphate groups are replaced by carboxy methyl groups and is glucosamine ring of lipid X moiety by a glucuronic acid ring in the case of TM2. This was done based on a combination of chemical, computational and biological considerations. Analogs of the type described here are synthetically challenging and their synthesis unprecedented because they involve carbon-carbon connectivities which involve difficulty to achieve stereochemical and logistical constraints. In this study, key intermediates in the synthesis of TM2, l-O-allyl-4-O-methyl- a,B-glucuronic acid benzylester (7), 2-acetamido-l-C—ethoxycarbomethyl-1,2-dideoxy-3- O—trimethylacetyl-4,6-O—benzylidene-B—D-glucopyranose (4) and (R)-3-hydroxytetradecanoic acid (8) were synthesized. These molecules and the associated synthetic methodology developed here should make the synthesis of a variety of structural analogs of lipid A possible. Structurally non-related lipid A antagonists TM3 and TM4 were also designed and synthesized to examine the roles of the charged head groups and lipid chains in the biological activities of lipid A at the molecular level, by considering these aspects separately without the saccharide components. Dedicated to my Mom, Dad, Youngah, Youngho and most importantly My Wife Mee-Sook iv ACKNOWLEDGMENT First, I would like to thank Dr. Rawle I. Hollingsworth ‘for his advise, support and the freedom to pursue my own idea. RIH, I owe you too much to describe here in detail. Thank you again to RIH. I would also like to thank the members of my guidance committee, Dr. Mlliam Reusch, Dr. Daniel Nocera and Dr. John McCracken, for their help and suggestions. I would like to bestow my gratitude to Robert & Deborah Elghanian who helped myself and family stay here enjoyably, more importantly, I would like to thank them for their friendship. I would also like to thank all of my friends from the Hollingsworth lab, Ben, Chuck, Jeongrim, Luc, Rob, Seunho, Ying, Yuanda, I would like to thank those persons who attended “Tuesday Night Meeting” in organic chemistry, Dr. Farnum, Dr. Reusch, Dr. Jackson, Dr. Stille, Iyere, Greg, Paul, Ken, Art, John, ----- . Thank you to everybody. I learned very much there. I would also like to thank Dr. Sungho Kang for inspiring me to be interested in organic chemistry. Most importantly, my parents, Jonghwa & Youngae Kim, without their help, support and tolerance, this would have been overwhelming adventure. I would also like to thank my sisters, Aera and Heera, for their help and encouragement. Lastly, I want to thank my wife, Mee-Sook, for all her love, encouragement and patience during the past five years. Thank you all. K-I Kim. TABLE OF CONTENTS LIST OF FIGURES LIST OF SCHEMES LIST OF ABBREVIATIONS INTRODUCTION RETROSYNTHETIC ANALYSES RESULTS AND DISCUSSION 1. PREPARATION OF THE LEFT MOIETY OF TMl 2. PREPARATION OF THE RIGHT MOIETY OF TMl 3. PREPARATION OF THE LEFT MOIETY OF TMZ 4. THE SYNTHESIS OF (R)-3-Hydroxytetradecanoic acid 5. SYNTHESES OF TM3 AND TM4 CONSLUSIONS EXPERIMENTAL REFERENCES vi l6 16 20 23 27 33 40 47 75 Figure Figure Figure Figure Figure Figure Figure Figure Figure 89%.“??? LIST OF FIGURES The structure of Escherichia coli lipid A The structure of TMl The structure of TM2 Structures of TM3 and TM4 Reaction basis for the synthesis of 8 List of molecules targeted in this research Side products formed in the reaction of 10 with (HCHO)n or CH2(OMe)2 Conformational analysis for the acid catalized acetalization of the diol 10 The list of molecules synthesized in this study vii (ht/1&5) 15 37 39 42 Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme Scheme 9939?:5PPI‘ i-Ar-Ir—i—i—i—r-Ir-I \roxmaunr—e 18. LIST OF SCHEMES Retrosynthetic analysis of TMl Retrosynthetic analysis of TM2 Retrosynthetic analysis of 8 Retrosynthetic alalysis of TM3 and TM4 Synthetic approach for the synthesis of 3 Reactions tried for the synthesis of 3 Synthetic scheme for the synthesis of 4 The reaction of 5 with malonate anion Synthetic scheme for the synthesis of 7 The unsuccessful synthetic approach to 7 . Synthetic scheme for the synthesis of 8 . Another synthetic approach for the synthesis of 8 . Alternative trial of the Wittig reaction of 37 . Synthetic scheme for the synthesis of TM3 . Synthetic scheme for the synthesis of TM4 . The generalized scheme for the syntheses of chiral coumpounds . The future synthetic scheme intended to be done for the left moiety of T M2 The future synthetic scheme intended to be done for the right moiety of TM2 and the coupling of the left moiety with the right moiety to give TM2 viii 10 12 13 14 18 19 21 22 25 26 30 31 32 35 36 43 46 4A n-BuLi Bn Bz 13C-NMR 1 8-Crown-6 d 8 DMF DMSO 2D-NMR Et ether Hz Imid. in vacuo lH-NMR IDA LPS LIST OF ABBREVIATIONS 4 Angstrom Acetyl Allyl n-Butyl lithium Benzyl Benzoyl Carbon ( 13C) NMR 1,4,7,10,13,l6-Hexaoxacyclooctadecane Doublet Chemical shift N,N-Dimethylformamide Dimethyl sulfoxide Two dimensional NMR Ethyl Diethyl ether Hertz Imidazole In vacuum oven Proton NMR Lithium diisopropylarnide Lipopolysaccharide Multiplet ix Me NMR PDC pet. ether Ph PMP Pv TMl TM2 TM3 TM4 TMS Tr p-TsOH Methyl Nuclear Magnetic Resonance Spectroscopy Pyridium dichromate Petroleum ether Phenyl para-Methoxyphenyl Pivaloyl ( Trimethylacetyl ) Pyridine Quartet Rate of flow Singlet Triplet Tetrabutylammonium bromide Tetrabutylammonium fluoride Trifluoromethanesulfonyl Tetrahydrofuran Thin Layer Chromatography The first target molecule designed The second target molecule designed The third target molecule designed The forth target molecule designed Tetramethylsilane Trityl ( Triphenylmethyl ) para-Tolunesulfonic acid INTRODUCTION Gram-negative bacteria contain in their cell surface a toxic principle which causes fever and lethal shock in higher animals. This toxin is called endotoxin since it is firmly bound to the cells in living bacteria and released only after the cells are destroyed. Endotoxin exhibits not only undesired toxic activities (fever, lethal shock) but also beneficial ones such as stimulation of immune response ( strong leukocytosis ) and induction of a tumor-necrotizing factor ( causing hemorrhage of tumors ). These toxic and beneficial biological activities have been attributed to the lipopolysaccharide ( LPS ) moiety, which is a complex amphipathic molecule found in the outer membrane of gram negative bacterial cells. In 1954, Westphal and Luderitz isolated the lipophilic part of LPS by mild acidic hydrolysis and termed it “Lipid A”, whose structure is shown in Figure 1.1, 2 They observed that lipid A manifested most of the endotoxic activities of LPS. This extremely important discovery was neither fully nor unanimously accepted by endotoxin investigators at that time. Later this concept was unequivocally confirmed by Japanese colleagues, based on a total synthesis of Escherichia coli lipid A and a comparison of synthetic lipid A’s biological activity with that of natural lipid A.3 It is now generally accepted that the lipophilic terminal substructure of LPS ( lipid A) is, in fact, responsible for its immunopharmacological activities and the full induction of endotoxicity. Since LPS (endotoxin ) is the most powerful immunostimulant known to date, the full induction of endotoxicity is believed to be an over-reaction of the immune system, as manifested by changes of cardiovascular parameters and white blood cell count, 2 disseminated intravascular coagulation, and finally, multi-organ failure leading to irreversible shock and death. In 1983, the structure of the lipid A component of Escherichia coli LPS was determined completely by means of chemical and 2D-NMR methods4 to be a glucosamine B-(l’-6)-disaccharide 1,4’-diphosphate acylated at two hydroxyls ( 3 and 3’ ) and the two amino groups, as shown in Figure 1. By referring to the determined structure, numerous studies were done to determine the structural features of lipid A associated with its biological activities. OH 403130 N” HO ore,-2 Figure l The structure of Escherichia coli lipid A Generally, it was believed that the phosphate groups in lipid A were necessary for endotoxic activity, based on the fact that lipid A derivatives without phosphate groups in 1 and 4’ positions were completely inactive, whereas a monophosphate at either 1 or 4’ exhibited much weaker but still definite endotoxic activity.5’6 This result doesn’t mean necessarily that the phosphate group is the only charged group which can lead to the 3 manifestation of the endotoxic properties of lipid A. We thought that phosphate groups funtion primarily to provide hydrophilic and anionic sites on the head group of lipid A, as well as contributing to the conformation of lipid A; but were not involved in chemical processes which are involved in the response to LPS. Recently published results7 are very encouraging, since they indicate that the phosphate group can be substituted by a carboxymethyl group with retention of similar biological activity. Besides efforts to determine the structural requirements for lipid A to exhibit endotoxic activity, numerous studies have addressed the possibility of dissociating detrimental endotoxic properties from beneficial immunostimulatory effects at the molecular level by chemical modification of lipid A or its simpler analogues. Efforts to date have concentrated on the total synthesis of lipid A3’8 and its derivatives,6’7’8 and on analogues representing both the nonreducing9’10’11 and the reducing sugar moietyl’12 such as lipid X and lipid Y. The final goal of this study is the elucidation of the molecular basis of the biological properties of lipid A. As a first step to this goal, we designed the first target molecule TMl, as shown in Figure 2, primarily to investigate structural requirements (primarily phosphate substituents ) of lipid A for endotoxicity. One model is that the phosphate group might be actively transfered as part of the mechanism. While the synthetic approach to TMl is to be discussed in a later chapter, the difficulty of synthesizing TMl forced us to design the second target molecule TM2 as shown in Figure 3. Besides the investigation of structural requirements of lipid A, structurally non-related lipid A antagonists, TM3 and TM4, as shown in Figure 4, were also designed and synthesized; primarily based on the fact that the biological activity of lipid A is very sensitive to counter cationic species, since the aggregate structure of lipid A has been known to be more relevant to its biological activity than the actual head group structure. The mechanism by which TM3 and TM4 inhibit the endotoxicity of lipid A is 4 thought to be by membrane perturbation. Although the results of biological activity tests of TM3 and TM4 as endotoxin antagonists were more interesting13 than expected, only the synthetic methods for obtaining them will be discussed here. OH H02 0 NH MM NH COZH HO H TMl Figure 2 The structure of TMl HOZC mom 0 Me(:&/\ co NH 2H [-10 HO TM2 Figure 3 The structure of TM2 AM WWW/W I, O ....\\ , as. ....\\ , TM4 TM3 Figure 4 Structures of TM3 and TM4 RETROSYNTHETIC ANALYSES A retrosynthetic strategy for TMl is shown in Scheme 1. The basic strategy for the synthesis of TMl had to be quite different from that employed in the synthesis of Escherichia coli lipid A by Japanese colleagues?»6 because of the different substituents in 1 and 4’ positions of lipid A. In the synthesis of Escherichia coli lipid A, phosphorylations of the 1 and 4’ hydroxyl groups were performed after 3, 3 ’ esterifications and 2, 2’ amidations by fatty acids. In the synthesis of TMl, alkylations of the 1 and 4’ carbons with the synthon of the carboxy methyl group were planned prior to esterifications or amidations of 3, 3’ or 2, 2’ by fatty acids, since C-alkylations at 1 and 4’ were expected to be more difficult than O-alkylations of 1 and 4’ hydroxyl groups. The reactions planned for the transformation from 3 to 1 are selective deprotections of acetyl groups on the 2 nitrogen of 3 and the trimethylacetyl groups on the 3 oxygen of 3, and mild coupling conditions with fatty acids for esterification and amidation. Selective deprotection of the N -acetyl group of 3 can be performed by a known method,6 the treatment with Meerwein’s reagent, triethyloxonium tetrafluoroborate, followed by acid hydrolysis of the resultant imidate. The free carboxylic acid groups on 1 and 4’ positions of TMl should be developed at the last stage without perturbing other ester groups. Since benzyl esters can be selectively cleaved by catalytic hydrogenation, the transesterification of ethyl ester 3 to a benzyl ester, prior to coupling with fatty acid was necessary. Therefore the deprotection of the trimethylacetyl group was intended to be by treatment with NaOBn in BnOH to place a benzylester on the 4 position of 3. In order to get 1 and 2, obviously 3 and 4 are key 8 intermediates prior to coupling with the fatty acids. The synthetic approach to 3 and synthesis of 4 from 5 will be discussed in the next chapter. The retrosynthetic strategy for TM2 is different from the one for TMl only for the lipid X moiety as shown in Scheme 2. The transformation from 7 to 6 would be even easier than in the case of 3. The known method14 for the deprotection of allyl ethers using tristriphenylphosphinerhodiuma) chloride would be applied to the selective deprotection of allyl glycoside 6. Since 7 is the key intermediate in Scheme 2, the synthesis of 7 from B-D-glucose will be discussed in the next chapter. Another important key intermediate for TMl and TM2 is optically pure (R)—3-hydroxytetradecanoic acid, 8, which was chemically made by Yoshihara Izurni15 in 1980. However, the best optical purity obtained by this method was 85% enantiomer excess. In order to get 8 in 100% enantiomer excess, the synthetic route from carbohydrate became inevitable. What was recently discovered in our group as shown in Figure 5, made this more than possible. Structure 41 was designed and synthesized as the key intermediate in the synthesis of 8 as shown in Scheme 3, based on the reaction mechanism in Figure 5. The synthesis of 8 from B-D-glucose will be discussed in the next chapter. TM3 and TM4 were made from L—tartaric acid as shown in Scheme 4 because, in addition to endotoxin antagonist, the possibility of these molecules for chiral auxilliary was another concern in this research. At this point, the key intermediates for the synthesis of TMl and TM2, in addition to TM3 and TM4, which will be discussed in RESULTS AND DISCUSSION chapter, are summarized in Figure 6. Scheme 1 Retrosynthetic analysis of TMl 10 OBn BrtOZC O mom H O NH Meo COZBn NH Et02 O Pit/V O OMe C023 3 4 A Ad) 0 A 5 HN 11 Scheme 2 Retrosynthetic analysis of TM2 12 TM2 BnOZC Mm . O O OAll o E 6 ll Br! 02 C MeO O HO OH OAll czs HO O HO OH B-D- glucose 4 A AcO O A 5 l-lN OH D-maltose 13 s: a 41 Scheme 3 [3- D- -glucose Retrosynthetic analysis of 8 OH OH 1) NaOH, H202 O 2) H30+ l-IO Figure 5 (S)-3-hydroxy-y-butyrolactone Reaction basis for the synthesis of 8 14 0 HO OH R 0=~\‘“ o + + N N< >~N N< HN NH HO OH :> :1) Hozc‘ C02H L-tartaric acid TM3 TM4 10 Scheme 4 Retrosynthetic analysis of TM3 and TM4 15 OBrt EtO Ph/v o 2 O 3110 C OMe 2 O C Et MeO g o=1\NH 0=/< : O== A e 7 c1 MeOH, (79%) 2) PhCH(0M6)2 5 0=< 11 NHAC p-TSOH, (91%) I’ll/v /\\ 0 Ph 0 &We NaH, THF . Mo... NHAc P Cl, 897 NHAC 12 V ( 0) 13 OBn ' OMe = v OMe HCl, (75%) m P O NHAc Nl-IAc 14 15 EtOZC OB“ (EtO)2POCH2COZEt NaH, THF 15 ’ on O OMe + Isomers 16 NHAc EtOz OBn ' Isomers + WC 0 OMe —< (unisolable) NHAC H2, 10% Pd-C 17 Scheme 5 Synthetic approach for the synthesis of 3 l9 OBn H O OBn 0 o NaBH4 o NHAC (43% from 14) Nl-IAC 15 18 OBn TfO O K+ -CH(CO2EI)2 ‘ on 0M9 l8-Crown-6 ' No reaction. NHAc 19 (Me3Si)CH2C02Et TBAF the” 19 :— No 3 formed. (IV/[€3Si)CI‘12COZEt LDA then 19 > No 3 formed. Scheme 6 Reactions tried for the synthesis of 3 (T020, P yr (67%) ll— 20 2. PREPARATION OF THE RIGHT MOIETY OF TMl In the synthesis of 4, obviously the C-alkylation of 5 to give 20 is the key reaction as shown in Scheme 7. Though syntheses of N -acetylglucosamine C—glycoside have been reported-'3326 in the preparation of lipid A analogues, their use has been limited because multistep routes are required for their preparations. The C-alkylation of 5 by malonate anion in the presence of l8-crown-6 was very efficient and stereoselective compared to known methods. The only problem in this transformation to give 21 in two steps was the low yield ( 21% from 5 ), caused by the formation of side products whose R}: values in the TLC were close to 20. It has been reported6that the treatment of the bromo analogue of 5 ( bearing Br instead of Cl ) with pyridine at room temperature for 20 min. gave the oxazoline 5b in high yield ( 90% ). Although not all of side products were identified, the major one of them should probably be the oxazoline 5b as shown in Scheme 8. Due to the competitive path to yield 5b, it was necessary to maximize the effective concentration of the nucleophile ( malonate anion ) for the formation of 20 to dominate over that of 5b. Moisture in the reaction mixture also seemed to accelerate the formation of 5b. As a result, it was important to add the minimum amount of the solvent and keep the reaction mixture anhydrous. Even with the low yield, the efficiency and high stereoselectivity (uniquely B-isomer) made this reaction noteworthy.27 Reactions used for the transformation from 21 to 22 were identical to ones used for the synthesis of 12 in Scheme 5, except that EtOH instead of MeOH was used as solvent to reduce the possibility of exchange of the ethyl group of the ester function. The trimethylacetylation of 22 was successful to give 4 in high yield ( 80% ), when the standard method was applied unlike in the case of 12. The difference in reactivities of 12 and 22 to this method ( PvCl, Pyr. and Imid. ) is difficult to explain. 21 A o A A20 KH, CH2(COzEt)2 > AfM on (80%) NHAc COZEt 4 Scheme 7 Synthetic scheme for the synthesis of 4 22 A A O 0 A20 _—4 A20 ‘CH(c02Et)2 HN ml 5 o=< 0 5a y/o -H+ A 0 A20 AcO o C0215: A COZEt N Scheme 8 The reaction of 5 with malonate anion 23 3. PREPARATION OF THE LEFT MOIETY OF TM2 The synthetic scheme for the synthesis of 7 is shown in Scheme 9. Intermediate 23 was prepared from B-D— glucose by adopting a p-methoxybenzylidene acetal group instead of a benzylidene acetal group, because the selective removal of the standard benzyl ether formed by reductive cleavage of a benzylidene acetal was anticipated to be very difficult without affecting the allyl ether group. After benzoylation of 23, reductive cleavage was performed with NaCNBH3 and p—TsOH. But the only product formed was completely deprotected diol 24 even in the presence of starting material. It seemed that the reduction of the p-methoxybenzyl ether reduced was even faster titan that of the p-methoxybenzylidene acetal. Therefore the acidic hydrolysis of the p-methoxybenzylidene acetal to effect total deprotection was employed to give 24. Subsequent protection of the 6-position by tritylation, methylation of the 4-position and detritylation gave 26. The pyridinium dichromate ( PDC )28 method was applied to the oxidation of 26 and the result was successful. However, the Jones reagent29 didn’t work for this case and seemed to cleave the benzoate groups. Subsequent debenzoylation and benzyl ester formation of the oxidized product of 26 gave 7. Another synthetic approach to 7 was attempted as shown in Scheme 10. The advantage of this approach was that we could work with only one isomer through most steps of the synthesis, rather than working on a mixture of or and B isomers for every step in Scheme 9. However, the glycosidic bond cleavage of 30 was unfortunately very difficult in a variety of conditions (70% aqueous HCOZH, AllOH-BF3, AllOH-(p-TsOH), HF, AllOH-AcCl). Although there was no conversion in most conditions, a couple of cases gave undesirable results. When HF was applied to 30 at 0°C and 25°C for one hour, the result was a mixture of products at 0°C and no 24 conversion at -25°C. When 30 was treated with AllOH and AcCl, intermediate 31 was the major product with no desired product observed by TLC or NMR. Based on the latter result, it seemed that carbonyl oxygen of the carboxylate retarded the glycosidic bond cleavage of 30 by reducing the basicity the acetal oxygen of glycoside 30. Intermediate 7 was successfully prepared by Scheme 9. 25 HO HO& 1) AllOH, AcCl PMP/\T OH 2) p-(MeO)PhCHO OH OAll B-D-glucose P'TSOH, (34%) 23 1) B20. Pyr. H&§L TrCl, Pyr. no :: H(B)z > Hg 2 AcOH, H o z ) (67%) 2 A 082 OAll (76%) 032 OAll 24 25 l) AgzO, Mel, DMF 1)PDC, DMF ¥ HO ' MeO 2) H30”: AcOH, Ergo 3&1 OBz COan OH O All Scheme 9 2) BnOH, K2CO3 ¥ '- 3) BnOH, p-TSOH, (28%) Synthetic scheme for the synthesis of 7 26 H 1) p-(MeO)PhCHO PMP o “9, p-TsOH T 0 > 32 HO OMe 2 B Cl,P ., 54‘7 BzO e Methyl-a-D-glucopyranoside ) Z yr ( 0) 27 OM 8110 NMe3-BH3, p-TsOH Hg&&‘ 1) AgzO, Mel, DMF > z > (90%) BzO OMe 2) H2, 10% Pd—C, (71%) 28 “1&3, 1) PDC, DMF C°2Meo “e8 = “em 2 MeOH, -TsOH BZO OMe ) (66%) P 320 OMe 29 30 co 2A” 0 AllOH, AcCl MeO < 82 BzO OM e 31 Scheme 10 The unsuccessful synthetic approach to 7 27 4. THE SYNTHESIS OF (R)-3-Hydroxytetradecanoic acid The successful scheme for the synthesis of 8 is shown in Scheme 11. The p-methoxybenzylidene acetal 33 was prepared from B-D-glucose by the same method as described for 23 except using BnOH instead of AllOH for preparing the glycoside. Dibenzylation of 33 was successfully performed by the reaction with N all and BnBr to give 34 in high yield ( 95% ). For the transformation of 34 to 37, generally the same reactions were used as are described for the preparation of 26 in Scheme 9. One change was that NaH-Mel was used for the methylation of 36. This method could not be applied for the methylation of 25 due to anticipated migration of the benzoyl group under the strongly basic conditions. Subsequent detritylation of the methylated product of 36 gave 37. In order to make 40 from 37, two options were explored. One was a Witti g reaction between the ylide of bromide 43 and l-decanal, the other was a Wittig reaction between the ylide of l-iododecane and the aldehyde 38. Originally the latter one was chosen because the isomerization at C-5 of aldehyde 38 was anticipated under the reaction condition for ylide formation. However, as shown in Schemes 12 and 13, even the preparation of the phosphonium bromide of 46 was either difficult or not successful due to poor conversion and/or difficulty in isolating the product. Since, in general, the amount of phosphonium halide required is in excess of the counterpart aldehyde, using the phosphonium halide of 46 prepared through multisteps synthesis in excess to commercially available l-decanal turned out to be an extremely inefficient route, particularly with poor conversion of 46 to the phosphonium bromide. By the same reasoning, Scheme 12 designed to work on single isomer all the way rather than working on a mixture ( 0t and [3 anomers ) as in Scheme 11, was discarded even though the isolated yield of 45 was higher ( 28% ) than the case of Scheme 13. Fortunately, 28 the anticipated isomerization of 38 prepared from 37 by the known method did not occur at all during the preparation of 40. In addition to that, the preparation of the phosphonium iodide 39 was very successful giving almost quantititive yield ( 94% ). The reduction and debenzylation of 40 by palladium catalyzed hydrogenation was successfully done to give 41 in quantititive yield The degradation of 41 to give (R)-3-hydroxytetradecanoic acid 8, the key reaction in Scheme 11, was performed in a mixture ( 1 : 1 ) of 0.2 M aqueous NaOH solution and 0.2 M aqueous H202 solution at 80°C for 2 days. Although TLC indicated that all starting material 41 was consumed after the first 10 hours, no desired product 8 was observed by lH-NMR analysis. The intermediates formed during the first 10 hours were not identified, but converted slowly to the desired product 8 without giving the 0t,B—unsaturated tetradecanoic acid The successful result in this reaction introduces a very useful method for the syntheses of chiral compounds. 29 Schemell Synthetic scheme for the synthesis of 8 3O HO ”0&0“ BnOH,AcC1 “1&0“ p—(MeO)PhCHO HO HO OH (77%) “0 p-TsOH, (38%) B-D-glucose 32 OH OB“ o PMP/v0 0 NaH, THF, BnBr I)MP/vo 910 BnO OH 03“ (95%) 03“ OBn 33 34 AcOH, H20 H:L%:\ TrCl, Pyr Hl?n0 (84%) DE“ DE“ (94%) OBn OB“ 1) N aH, THF, MCI HO _ O ACZO, DMSO “’80 ——> 2) H3O+, ACOH, EIZO (54%) OB“ OBn (65%) 37 (CHflgCHa CHO + _ / W) 0 131131) (CH2)9CH3I (39) BnO MeO O OBn . BnO OBn n-Bqu, THF, (74%) QB 38 " OBn 40 (CH2)3CH3 H , 10% Pd—C NaOH, H O 2 M90 0 —22, HO2C /Y\ PMP M -TsOH, 480/) H HO OMe p ( 0 HO OMe Methyl-a-D-glucopyranoside 42 1) BzC1.Pyr. T O 1) AgZO, Mel, DMF 2) AcOH, H20 ng > 3)TrC1, pyr, (57%) 4 32° OMe 2) H3O+, AcOH, EtzO, (65%) 3 Br HO MeO O P P113. CBr4 MeO O PPh3, DMF 320 > 82 > 820 OMe THF, (86%) BzO OMe (24%) 29 44 P+Ph3 B!" o n-BuLi, THF ““320 > No desired product formed. BzO l-Decanal OMe 45 Scheme 12 Another synthetic approach for the synthesis of 8 32 I’il’h3 Br' n-BuLi, THF MeO O . 820 No desued product formed. B l-Decanal ZO OMe 45 Br HO MeO O PPh3, CBI‘4 M60 0 BnO —’ BnO OBn OBn (71%) OBn OBn 37 46 PPh3, DMF Poor conversion. Difficult to purify. Scheme 13 Alternative trial of the Wittig reaction of 37 33 5. SYNTHESES OF TM3 AND TM4 TM3 was prepared from L-tartaric acid by Scheme 14. Tartaric acid diamide 10 was prepared in two steps through dimethyltartrate. Although the one step coupling of L-tartaric acid with l—decylamine was attempted with DCC, ammonolysis of the diester turned out to be the best way because of the easy work. N 0 further purification was needed. 47 was prepared in excellent yield and used in the next step without purification. Reduction of 47 was performed by lithium aluminium hydride to give 48. Methylation of 48 was performed in heterogeneous solution to give TM3 in poor yield ( 33% ), but, because of the isolation procedure, the purity of the product was so high that no further isolation or purification step was needed after washing the reaction mixture once with water. TM4 was prepared by the same reaction sequences as TM3 except for one step as shown in Scheme 15. 49 could not be made by any acid catalyzed acetalization ( e. g. formalin-HG], paraformaldehyde-HCI, CH2(OMe)2-(p-TSOH), CH2(OMe)2-BF3, CH2(OMe)2-P205 ). In acid catalized reactions for the synthesis of 49, many of side products were formed and some of them were identified as shown in Figure 7 based on 1I-INMR spectra. Structures of 51, 52 and 53 gave rise to the quite interesting question of how can the small difference in similar reagents ( CH2(OMe)2 and Me2C(OMe)2) made have such different effects on the results of the transacetalization of 10. Based on the structures in Figure 7, the equilibrium between conformations of 10, in addition to the difference in reagents, seemed to cause quite different results in the syntheses of 47 and 49. In the reaction of 10 with Me2C(OMe)2 ( bp 83°C ), the byproduct MeOH ( bp 64.6°C ) was removable by fractional distillation to drive the equilibrium to the formation of the product. However, in the reaction of 10 with CH2(OMe)2, the higher boiling point ( 646°C ) of MeOH compared to 42°C for the reagent CH2(OMe)2 made it impossible 34 for the equilibrium to be driven to the formation of the product 49 by this method Another important factor should be considered in explaining the formation of the side products 51, 52 and 53. The side product 51 was formed when the diol 10 was treated with the reagent paraformaldehyde. The other side products 52 and 53 were formed when the diol 10 was treated with CH2(OMe)2 in various acidic conditions. The structures of 51, 52 and 53 indicated that the intramolecular cyclization to form five- membered acetal ring was not favorable at all in these cases. The conformational analysis of the diol 10 seemed to provide a reasonable rationalization of these results. There are three possible conformers for the diol 10. There are CFl, CF2 and CF3 as shown in Figure 8. CFl seemed the most stable conformer compared to CF 2 or CF3, if one considers the H-bonding between hydroxyl and amide groups, and the lipophilic interaction between the C-14 alkyl chains. The two hydroxyl groups of the diol 10 were anti to each other in the conformer CF 1, so that compound 51 with the seven—membered ring would have had to be formed when the diol 10 was treated with paraformaldehyde even with the removal of the byproduct ( water). In the case of the reagent CH2(OMe)2, the presence of the byproduct MeOH in the reaction mixture, seemed to prevent the formation of the desired product 49 which would have been formed through the (unfavorable) conformer CF2 or CF3. As soon as the desired product 49 was formed, the byproduct MeOH seemed to cleave 49 and drive the equilibrium to the favorable conformer CFl. Therefore, treatment with the reagent CH2(OMe)2 did not lead to intramolecular cyclization in acidic conditions. On the other hand, once the product 49 was formed, the reagent CH212 with a base K2CO3 gave no chance for the product 49 to be cleaved by the byproduct (potassium iodide ), even though ( also in the basic reaction condition ) the product 49 had to be formed through either conformer of CF2 or CF3 . These results seemed to provide some structural information about how the long alkyl chains should be aligned in the molecule. 35 HO OH HO OH 1 M H .. ) pr‘gOH o=e O (MeO)2C(CH3)2 H026 COZH 4’ HN NH T OH (1007 ) VF . - s , L- tartaric acid 2) 1-Tetradecylamrne g p 0 (97%) R R (R = “(CH2)11CH3) 10 I I I e r. I O O , LiAlH4 Lg K2C03, TBAB «ngcr—gso; °.. 0 $. > i + + or“ N; (77%) is: N; (CH3)ZSO4,(33% >N >< R$ R R R R R 47 48 TM3 Scheme 14 Synthetic scheme for the synthesis of TM3 36 HO OH \__8: GAO K CO , CH I LiAlH o=-‘\ o 2 3 2 2 _ 0%ng o 4 7 § N211 18-Crown-6, (89%) i” I? (79%) R R 10 (R = '(CH2)11CH3) R 49 R /\ 0A0 O 0 Pg ch0;,, TBAB Lg 29,3504- 1w NH (CH3)ZSO4, (45%) >513 i1< R3 3R R R 50 TM4 Scheme 15 Synthetic scheme for the synthesis of TM4 ¥ 7 37 / \ \ o o 0 [a0 o> MeO O OBn BnO 0"“ n-BuLi, THF can 08“ R H2, 10% Pd—C NaOH, H o = M e0 0 2 2 > HOZC/Y\CH2R MeOH HO 0H OH OH Scheme 16 The generalized scheme for the syntheses of chiral compounds from B-D-glucose TsOZC /Y\(CH2)9CI‘13 ( S4 ) OBn / ( For the right moiety 4 ) HOZC (012)9CH3 OH Ts C 8 o (CI—{2)9CI'I3 (For the left moiety 7 ) Bn02C Meow O O o O X BHOZC O HQ :> O 0” OAll 56 7 (x = C1 or Br or OCNHCH3) Scheme 17 The future synthetic scheme intended to be done for the left moiety of TM2 45 Scheme 18 The future synthetic scheme intended to be done for the right moiety of TM2 and the coupling of the left moiety with the right moiety to give TM2 46 rah/T00 o TrO o O 54 MeO COzEt => 0 O NH §< o=< 4 BnO (57 ) 54 Ho MeO O 0 co B 0 NH 2 n o BnO BnO 2 56 HOZC 0 Memo 0 o o MeO 0 w 0 COZH o o 0 NH 0 0 Ho Ho TM2 47 EXPERIMENTAL 1H-NMR and l3C-NMR spectra were measured on a Varian Gemini-300 spectrometer ( 300 MHz ) for chloroform-d solutions unless noted otherwise. The chemical shifts are given in 5 values with TMS as the internal standard or relative to the chloroform line at 7.24 ppm for proton or 77.4 ppm for 13C spectra. Silica gel flash column chromatography was carried out on silica using Kieselgel 60 ( Merck), 0.040- 0.063 mm. Precoated Kieselgel F254 plates ( 1mm thickness ; Uniplate ) were used for preparative TLC. The intermediates 5 and 11 in Scheme 5 were prepared by an earlier developed method13 from B-D-glucosamine hydrochloride. Methyl 2-acetamido-4,6-O-benzylidene-Z-deoxy-B-D-glucopyranoside (12) Compound 11 ( 30g, 82.9 mmol ) was treated with potassium carbonate ( 0.5g, 3.6 mmol ) and methanol ( 200 ml ) at room temperature for 16 hours. After neutralization with 35% aqueous hydrochloric acid ( 0.5 g, 5 mmol ), insoluble materials were filtered off. The filtrate solution was concentrated under reduced pressure and the residue was dried in vacuo. This residue was dissolved in dry DMF ( 100 ml ) and treated with benzaldehyde dirnethylacetal ( 18.9g, 124 mmol ) and p-TsOH ( 0.2g, 1 mol) at room temperature for 2 hours. After evaporating at reduced pressure, potassium carbonate (0.3g, 2.2 mmol ) was added. After additional stirring for 0.5 hour, insoluble materials were filtered off and washed with DMF (20 ml ). The filtrate was combined with the washings and evaporated at reduced pressure. The residue was then treated with ether ( 150 ml ) to precipitate the product. The product was crystalized 48 from ether, collected by filtration, washed with distilled water ( 50 ml ) and ether ( 50 ml ) and then dried in vacuo. Yield; 24.4g ( 91% ). 1H-N MR ( 300 MHz, CDClg-CD3OD ) 8 2.02(3H, s, CH3), 3.51(3H, s, CH3), 3.40-3.55(2H, m, H—5 and H-6ax), 3.62(1H, dd, J = 9 and 10 Hz, H-4), 3.77(1H, dd, J = 10 and 10 Hz, H-3), 3.89(1H, dd, J = 9 and 10 Hz, H-2), 4.35(1H, dd, J = 10 and 6 Hz, H-6eq), 4.54(1H, d, J = 9 Hz, H-l), 5.S7(1H, s, H-benzyl), 7.34—7.53(5H, m, H-aromatic). Methyl 2-acetamido-3-O-trimethylacetyl-4,6-O-benzylidene-Z-deoxy-B-D- gluc0pyranoside (13) To a solution of 12 ( 19.2g, 59.4 mmol ) in dry THF ( 300 ml ) was added 60% N all in mineral oil ( 2.7g, 67.5 mol) at room temperature. After stirring the THF solution at room temperature for 18 hours, it was cooled to 0°C and trimethylacetyl chloride (7.2g, 59.4 mmol ) was added keeping the solution at 0°C. The reaction mixture was allowed to be warmed up to room temperature and stirred for another 4 hours. The reaction mixture was then concentrated under reduced pressure. Dichloromethane ( 300 ml ) and distilled water ( 100 ml ) were added to the residue. The organic layer was separated in a separatory funnel, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The crude oily product was purified by flash column chromatography ( CHzClz-ether : 1-1) Yield ; 21.5g ( 89% ). 1H-NMR ( 300 MHz, CDC13 ) 5 l.18(9H, s, (CH3)3), 1.93(3H, s, CH3), 3.25(3H, s, OCH3), 3.57(1H, m, H-5), 3.70(1H, dd, J = 10 and 10 Hz, H-6ax), 3.74(1H, dd, J = 10 and 10 Hz, H-4), 4.15-4.30(3H, m, H-1 H-2 and H-6eq), 5.28(1H, dd, J = 10 and 10 Hz, H-3), 5.50(1H, s, H-benzyl), 7.29-7.40(5H, m, H-aromatic). 13C-NMR ( 75.5 MHz, CDC13) 8 101.0(C-1), 170.5(C-carbonyl), 180.0(C—carbonyl). 49 Methyl 2-acetamido-3-O-trimethylacetyl-6-O-benzyl-2-deoxy-B-D- gluc0pyranoside (14) A solution of the benzylidene acetal 13 ( 0.7g, 1.7 mmol ) and sodium cyanoborohydride (0.9g, 14.0 mmol ) in dry THF ( 20 ml ) containing 4A-molecular sieves was cooled to 0°C. Anhydrous hydrogen chloride was bubbled into the reaction mixture until no gas evolution. After 10 min at 0°C, when TLC indicated complete reaction, the mixture was poured into ice water, and the product was extracted with dichloromethane ( 50 ml ). The extract was washed with saturated aqueous sodium hydrogencarbonate, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was subjected to flash column chromatography ( CH2C12-MeOH : 20-1 ). Yield ; 0.53g (75% ). 1H-NMR ( 300 MHz, CDC13 ) 8 l.19(9H, s, (CH3)3), 1.92(3H, s, CH3), 3.47(3H, s, OCH3), 3.57(1H, m, H-5), 3.73(1H, dd, J = 10 and 10 Hz, H-4), 3.78(2H, d, J = 5 Hz, H-6), 4.03(1H, dd, J = 10 and 8 Hz, H—2), 4.38(1H, d, J = 8 Hz, H-l), 4.56(1H, d, J = 12 Hz, H-benzyl), 4.62(1H, d, J = 12 Hz, H-benzyl), 5.05(1H, dd, J = 10 and 9 Hz, H-3), 7.33(5H, m, H-aromatic). l3C-NMR ( 75 .5 MHz, CDC13) 5 102.0(C-1), 170.4(C-carbonyl), 180.0(C-carbonyl). Methyl 2-acetamido-3-O-trimethylacetyl-4-oxo-6-O-benzyl-2-deoxy-B-D- gluc0pyranoside (15) To the solution of anhydrous DMSO ( 500 ml ) and acetic anhydride (25 ml ) was added 14 ( 2.5g, 6.1 mmole ). The reaction mixture was stirred at 30°C for two days and concentrated by vacuum distillation at 30°C. The oily residue was used for the next reaction without further purification. 50 Methyl 2-acetamido-3-O-trimethylacetyl-cis,trans-4- ethoxycarbomethylene-6-O-benzyl-2,4-dideoxy-B-D-gluc0pyranoside (16) To the suspension of 60% N aH in mineral oil ( 264 mg, 6.6 mmole ) and dry THF ( 10 ml) was added ethyl diethylphophonoacetate ( 1.30g, 6.1 mmole ) at 0°C and the reaction mixture was stirred at room temperature for 3 hours. The ketone 15 was added to the reaction mixture which was stirred at room temperature for additional 16 hours. After checking the completion of the reaction by TLC, the reaction mixture was concentrated and the residue‘was treated with dichloromethane ( 50 ml ) and distilled water (20 ml ). The organic layer was separated in a separatory funnel, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. Reaction products were subjected to flash column chromatography and analyzed by 1H- NMR. Methyl 2-acetamido-3-O-trimethylacetyl-6-O-benzyl-Z-deoxy-B-D- galactopyranoside (18) The crude oxidized product 15 from 14 ( 2.0g, 4.89 mole ) was treated with NaBH4 (0.5g, 13.2 mmole ) and TBAB (2 mg) in dichloromethane ( 15 ml) and distilled water ( 5 ml). After stirring for 16 hours, the completion of the reaction was checked by TLC and the organic layer was separated, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The oily residue was subjected to flash column chromatography ( CH2C12-MeOH : 20—1 ). Yield; 850 mg ( 43% ). TLC ; R}: = 0.25 ( CH2C12-MeOH : 20-1 ). lH-NMR ( 300 MHz, CDC13 ) 8 l.21(9H, s, (CH3)3), 2.03(3H, s, CH3), 3.52(3H, s, OCH3), 3.48-3.60(2H, m, H-6), 3.74-3.85(2H, m, H-2 and H-S), 3.91(1H, dd, J = 11 and 3 Hz, H-3), 4.41(1H, d, J = 10 Hz, H-l), 4.47(1H, d, J = 12 Hz, H-benzyl), 4.53(1H, d, J = 12 Hz, H-benzyl), 5.33(1H, dd, J = 3 and 1 Hz, H—4), 7.25-7.36(5H, 51 m, H-aromatic). 13C-NMR (75.5 MHz, CDC13) 8 101.5(C-l), 173.5(C-carbonyl), 178.0(C-carbonyl). Methyl 2-acetamido-3-O-trimethylacetyl-4-O-trifluoromethanesulfonyl-6- O-benzyl-Z-deoxy-B-D-galactopyranoside (19) To the solution of 18 ( 90 mg, 0.22 mmole ) in dry pyridine ( 2 m1 ) was added trifluoromethanesulfonic anhydride ( 100 mg, 35 mmole) at —15°C. The reaction mixture was warmed up to room temperature and stirred for 2 hours. It was then treated with 4% aqueous HCl solution ( 35 m1) and dichloromethane ( 20 ml ). The organic layer was separated and washed with a 5% aqueous solution of NaHCO3 ( 10 ml ). The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The oily residue was subjected to flash column chromatography (CH2C12-MeOH : 50-1 ). Yield ; 80 mg ( 67% ). TLC; R1: = 0.30 ( CH2C12—MeOH : 50-1 ). 1H-NMR ( 300 MHz, CDC13 ) 8 l.l7(9H, s, (CH3)3), 2.12(3H, s, CH3), 3.53(3H, s, OCH3), 3.45-3.58(2H, m, H-6), 3.71(1H, ddd, J = 10, 10 and 4 Hz, H-2), 4.10(1H, ddd, J = 10, 10 and 2 Hz, H-5), 4.43(1H, d, J = 12 Hz, H-benzyl), 4.51(1H, d, J = 12 Hz, H-benzyl), 4.56(1H, d, J = 10 Hz, H-l),5.01(1H, dd, J = 4 and 2 Hz, H-4), 5.13(1H, dd, J = 4 and 4 Hz, H-3), 5.67(1H, d, J = 10 Hz, NH), 7.24-7.36(5H, m, H-aromatic). 13C-NMR ( 75.5 MHz, CDC13) 8 99.5(C—1), 119.4(q, J = 320 Hz, CF3), 168.7(C—carbonyl), 176.5(C-carbonyl). Ethyl 2-acetamido-l,2-dideoxy-3,4,6-tri-O-acetyl-B-D- glucopyranosylacetate (21) To a 60% mineral oil dispersion of KH ( 2.67g, 40 mole ), was added anhydrous dichloromethane (20 m1). Diethyl malonate ( 6.4g, 40 mole ) and 52 18-crown-6 ( 5.3g, 20 mole ) were then added to the suspension while maintaining it at 0°C. After additional stirring at 0°C for 15 min, a-D—acetochloroglucosamine 5 ( 3.66g, 10 mole ) was added in one portion. After additional stirring at room temperature for 20 min, acetic acid ( 2.4g ) was added to quench the reaction. The solution was washed with 5% aqueous sodium hydrogencarbonate ( 100 ml), dried over anhydrous magnesium sulfate, filtered and concentrated The product was purified by flash column cchromatography ( CH2C12—MeOH : 30-1 ). The first purified oily product 20 was dissolved in DMSO (24 ml ) containing N aCl ( 2.0g, 34 mole ). After refluxing for 16 hours, DMSO was removed by vacuum distillation and the residue was treated with distilled water ( 50 ml ) and dichloromethane ( 100 ml ). The organic layer was isolated, dried over anhydrous magnesium sulfate, filtered and concentrated The product was purified by flash column chromatography ( CH2C12-MeOH : 30-1 ). Yield; 0.88g ( 21% ). TLC; R}: = 0.35 (CH2C12-MeOH : 20-1 ). [001320 ; -1.94 ( c 0.09, CHC13 ). 1H-NMR ( 300 MHz, CDC13 ) 8 1.23(3H, t, J=7 Hz, CH3), 1.90(3H, s, CH3), 2.00(3H, s, CH3), 2.01(3H, s, CH3), 2.04(3H, s, CH3), 2.58(2H, d, J=6.0 Hz, CH2), 3.62(1H, m, H-5), 3.79(1H, dt, J=10 and 6 Hz, H-l), 4.01-4.22(4H, m, H-2, H-6 and OfflzCH3), 4.98(1H, dd, J=10 and 10 Hz, H-4), 5.06(1H, dd, J=10 and 10 Hz, H-3). l3C-NMR ( 75.5 MHZ, CDCl3) 8 37.7(C—2). MS (Neg. FAB, matrix: NBA) 416.2(M-1). 2-Acetamido-l-C-ethoxycarbomethyl-1,2-dideoxy-4,6-O-benzylidene-B-D- gluc0pyranose (22) The procedure for the preparation of 22 was the same as that employed for the preparation of 12, except that EtOH was used instead of MeOH Yield; 55% . TLC; RF = 0.20 ( CH2C12-MeOH : 20-1 ). 1H-NMR ( 300 MHz, CDC13 ) 8 1.25(3H, t, J=7 Hz, CH3), 1.94(3H, s, CH3), 2.55(2H, m, CH2), 3.36-3.92(5H, m, H-2, H-3, H-4, H-5 and H-6), 53 4.14(2H, q, J = 7 Hz, OCHz), 4.26(1H, dd, J = 10 and 4 Hz, H-6), 5.47(1H, s, H-benzyl), 7.33-7.48(5H, m, H-aromatic). 2-Acetamido-l-C-ethoxycarbomethyl-1,2-dideoxy-3-O-trimethylacetyl- 4,6-O-benzylidene-B-D-glucopyranose (4) To the solution of imidazole (0.6g, 8.8 mmole ) in dry pyridine ( 12 ml ) in an ice bath, was added trimethylacetyl chloride ( 1.0g, 8.3 mmole ). After removing ice bath, the solution was stirred at room temperature for 0.5 hour. Compound 22 ( 1.8g, 3.9 mmole ) was added to the solution and it was stirred at 80°C for 16 hours. After evaporating pyridine under reduced pressure, the residue was treated with dichloromethane ( 50 ml ) and 5% aqueous HCl solution ( 20 ml ). The organic layer was isolated and washed with 5% aqueous NaOH solution ( 10 ml ). The organic layer was collected, dried over anhydrous magnesium sulfate and concentrated. The product was purified by flash column chromatography ( CH2C12-MeOH : 20-1 ). Yield ; 1.7g ( 80% ). TLC; R1: = 0.30 ( CH2C12-MeOH : 20-1 ). lH-NMR ( 300 MHz, CDC13 ) 8 1.19(9H, s, (CH3)3), 1.25(3H, t, J=7 Hz, CH3), 2.51(1H, dd, J = 18 and 10 Hz, CHI-1’), 2.62(1H, dd, J = 18 and 5 Hz, CHH’), 3.49(1H, m, H-l), 3.69(2H, m, H-5 and H-6), 3.86(1H, m, H-2), 4.12(1H, dd, J = 10 and 10 Hz, H-2), 4.15(2H, q, J = 7 Hz, OCHz), 4.24(1H, dd, J = 10 and 5 Hz, H-6), 5.17(1H, dd, J = 10 and 10 Hz, H-3), 5.52(1H, s, H-benzyl), 7.33-7.43(5H, m, H-aromatic). Allyl 4,6-(4-methoxy)benzylidene-0t,B-D-glucopyranoside (23, a/B=3/2) To a solution of B-D-glucose ( 60g, 0.33 mole ) and allyl alcohol ( 300 ml ) was added acetyl chloride ( 0.5 g, 6.4 mmole ). The reaction mixture was refluxed for 18 hours and concentrated under reduced pressure after checking for completion by periodically obtaining 1H-NMR spectra in D20. Dry DMF ( 100 ml), p-TsOH ( 0.5g ) 54 and p-anisaldehyde ( 45.3 g, 0.33 mole) were then added to the residue. After stirring the reaction mixture at room temperature for 2 hours, toluene ( 200 ml ) was added and it was then evaporated under reduced pressure. This addition-evaporation step with toluene was repeated at least three times. Anhydrous potassium carbonate ( 1.0g ) was then added to the reaction mixture and it was stirred for 0.5 hour at room temperature. It was then concentrated in vacuo and the residue treated with ether (250 ml ) The ethereal was allowed to stand in the refrigerator ( -20°C ) for 18 hours. The precipitate which formed was collected by filtration and the filter cake was washed with a 1:1 mixture ( 200 ml ) of pet. ether and ether. The filter cake was redissolved and the product isolated by flash column chromatography ( CH2C12-MeOH : 20-1 ). Yield ; 38g ( 34% from D-glucose). TLC ; R1: = 0.33 ( CH2C12-MeOH : 20-1 ). 1H—NMR ( 300 MHz, DMSO-d6 ) 8 3.12(1H, ddd, J = 10, 10 and 7 Hz, H(B)-6), 3.32-3.40(1H, m, H(0t)-6), 3.58-3.69(2H, m, H-3 and H-4), 3.73(3H, s, OCH3), 3.93-4.27(3H, m, H-5 and H(0t)-allyl), 4.37(1H, d, J = 10 Hz, H(B)-1), 4.78(1H, d, J = 4 Hz, H(0t)-1), 4.99(2H, d, J = 8 Hz, H(B)-allyl), 5.13-5.19(2H, m, H-2 and H(B)-vinyl), 5.27-5.39(2H, m, H(0t)-vinyl), 5.50(1H, s, H-benzyl), 5.83-6.00(1H, m, H-vinyl), 6.90(2H, d, J = 10 Hz, H-aromatic), 7.36(2H, d, J = 10 Hz, H-aromatic). Allyl 2,3-di-O-benzoyl-a,B-D-glucopyranoside (24, alB=3/2) To a cooled solution of 23 ( 18g, 53 mole ) in dry pyridine ( 50 ml ) in an ice bath was added benzoyl chloride ( 15g, 106 mmole ). The reaction mixture was then stirred at room temperature for 18 hours and concentrated under reduced pressure. Dichloromethane( 100 ml ) was added to the residue and the solution was washed with a 4% aqueous HCl solution ( 100 ml ) followed by a 5% aqueous NaHCO3 solution ( 50 ml ). The organic layer was isolated, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was treated with an 55 AcOH—H20 ( 5 : 1 ) mixture ( 100 ml ) at room temperature for 16 hours. The reaction mixture was then concentrated under reduced pressure until 80% of the solvent was evaporated. A 5% aqueous N aHCO3 solution was added to the concentrated reaction mixture until no further gas evolution. The product was then extracted from the aqueous suspension by washing twice with dichloromethane ( 100 ml ). The combined organic solution was dried over anhydrous magnesium sulfate, filtered and concentrated. The product was purified by flash column chromatography ( CH2C12-MeOH : 20-1 ). Yield ; 15.3g ( 67% from 23 ). TLC ; R}: = 0.30 ( CHzClz-MeOH : 20-1 ). 1H—NMR ( 300 MHz, CDCl3 ) 8 3.56(1H, m, H(B)—5), 3.85-4.02(3H, m, H-4, H(0t)-5 and H—6), 3.98-4.36(2H, m, H-vinyl), 4.76(1H, d, J = 8 Hz, H(B)—l), 5.08-5.30(4H, m, H(0t)-1, H-2 and H(0t)-allyl), 5.38-5.47(2H, m, H(B)-allyl), 5.69-5.89(2H, m, H-3 and H-vinyl), 7.30—7.96(10H, m, H-aromatic). 13C—NMR ( 75.5 MHz, CDC13) 8 95.5 (Ca-l), 100.0(CB-1), 118.0(C-allyl). Allyl 2,3-di-O-benzoyl-6-O-triphenylmethyl-a,B-D-glucopyranoside (25, oc/ B=3/ 2) To the solution of 24 ( 8.7g, 20.3 mmole ) in dry pyridine (20 ml ) was added triphenylmethyl chloride ( 6.3g, 22.3 mmole ). The reaction mixture was stirred at room temperature for 16 hours. Dry ether ( 150 ml ) was then added and stirring was continued for a further 4 hours. The reaction mixture was then washed with distilled water ( 50 ml ) and 4% aqueous HCl solution ( 200 ml ). The organic layer was collected and washed with 5% aqueous NaHCO3 solution ( 50 ml ). After drying it over anhydrous magnesium sulfate, the organic layer was concentrated under reduced pressure. The residue was subjected to flash column chromatography (pet. ether—CHgClz : 1-1 ). Yield ; 10.4g ( 76% ). TLC ; R}: = 0.27 ( pet. ether-CH2C12 : 1-2 ). 1H—NMR ( 300 MHz, CDC13 ) 8 3.45-3.68(2H, m, H-5 and H-6), 3.90—4.44(4H, m, H-4, H-6 and H-vinyl), 4.27(1H, d, J = 10 Hz, H(B)-1), 5.15-5.50(3H, m, H(0t)-1, 56 H-2 and H-allyl), 5.78-5.96(2H, m, H-3 and H-vinyl), 7.25-8.04(25H, m, H-aromatic). 13C-NMR (75.5 MHz, CDC13) 8 117.8(C—a11y1). Allyl 2,3-di-O-benzoyl-4-O-methyl-0t,B-D-glucopyranoside (26, alB=3/2) To the solution of 25 (4.3g, 6.4 mmole ) containing AgzO ( 2.97g, 12.8 mmole ) in dry DMF ( 14 ml ) at room temperature was added methyl iodide ( 1.82g, 12.8 mmole ). The reaction mixture was stirred at room temperature for 18 hours and then filtered through celite after the addition of ether ( 100 ml ). The filtrate was washed with distilled water ( 100 ml ) and the organic layer was isolated and dried over anhydrous magnesium sulfate. The drying agent was removed ty filtration and the filtrate was concentrated and the residue was subjected to flash column chromatography ( pet. ether-CH2C12 : 1-2 ). The purified product was treated with ether ( 20 ml ), acetic acid ( 10 ml) and 10% HCl aqueous solution (5 ml ). After stirring the reaction mixture at room temperature for 18 hours, only the ether was removed from the reaction mixture by evaporation under reduced pressure. After adding dichloromethane ( 120 ml ) to the concentrated mixture, 5% aqueous NaHCO3 solution was added until no further gas evolution. The organic layer was separated, dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was subjected to flash column chromatography ( CH2C12-MeOH : 20-1 ). Yield ; 1.7g ( 64% ). TLC ; R1: = 0.40 ( CHzClz-MeOH : 20-1 ). 1H-NMR ( 300 MHz, CDCl3 ) 5 3.45(3H, s, OCH3(0t or 13)), 3.47(3H, s, OCH3 (a or 13)), 3.51-3.57(1H, m, H-S), 3.68(1H, dd, J = 10 and 10 Hz, H—4), 3.82-4.38(4H, m, H-vinyl and H-6), 4.77(1H, d, J = 9 Hz, H(B)-1), 5.10-5.32(4H, m, H(0t)-1, H(0t)-2 and H-allyl), 5.36(1H, dd, J = 10 and 9 Hz, H(B)-2), 5.65(1H, dd, J = 10 and 10 Hz, H(B)-3), 5.71-5.89(1H, m, H-vinyl), 6.00(1H, dd, J = 10 and 10 Hz, H(0t)-3), 7.34-8.05(10H, m, H-aromatic). 57 1-O-Allyl-4-O-methyl-oc,B-D-glucuronic acid benzylester (7, Ot/[3=3/2) Pyridinium dichromate ( 5.0g, 13.3 mmole ) was added to a solution of 26 ( 1.7g, 3.8 mmole ) in dry DMF( 10 ml ) at room temperature. After stirring at room temperature for 18 hours, distilled water ( 100 ml ) and dichloromethane were added to the reaction mixture. The organic layer was separated, dried over anhydrous magnesium sulfate, filtered and concentrated. The concentrate was subjected to flash column chromatography ( CHzClz-MeOH: lO-l ). The product was collected and treated with benzyl alcohol ( 30 ml ) and potassium carbonate ( 2.0g ). After stirring at room temperature for 18 hours, the reaction mixture was filtered and p-TsOH was added to the filtrate until the solution was strongly acidic (pH < 2 on pH-paper ). The solution was then stirred at room temperature for 18 hours and coevaporated with toluene ( 100 ml ) under reduced pressure. Coevaporation with toluene was repeated three times, and the solution containing benzyl alcohol was treated with sodium bicarbonate (2g). After stirring for half an hour, it was filtered and subjected to flash column chromatography ( CHzClz ) to remove most of the benzyl alcohol in the fore-run. The product which was still adsorbed on the silica gel on the column was then eluted sith a more polar solvent system ( CH2C12-MeOH : 20-1 ). Yield ; 0.37g ( 28% from 26 ). TLC ; RF = 0.33 ( CHgClg—MeOH : 20-1 ). 1H-NMR ( 300 MHz, CDC13 ) 8 3.33(3H, S, OCH3(0t or 13)), 3.35(1H, dd, J = 10 and 10 Hz, H(0t)-3), 3.36(3H, S, OCH3(0t or 13)), 3.45(1H, dd, J = 10 and 9 Hz, H(B)-2), 3.56(1H, dd, J = 10 and 4 Hz, H(0t)-2), 3.62(1H, dd, J = 10 and 10 Hz, H([3)-3), 3.80(1H, d, J = 10 Hz, H(B)-5), 3.82(1H, dd, J = 10 and 10 Hz, H-4), 4.00—4.40(2H, m, H-vinyl), 4.14(1H, d, J = 10 Hz, H(0t)-5), 4.32(1H, d, J = 10 Hz, H(B)-1), 4.95(1H, d, J = 4 Hz, H(0t)-l), 5.19-5.30(4H, m, H-allyl and H—benzyl), 5.80-5.95(1H, m, H-vinyl), 7.32-7.38(5H, m, H-aromatic). l3C-NMR ( 75.5 MHz, CDC13) 8 80.6(Ca or B-benzyl), 80.9( Ca or fi-benzyl), 97.4(Ca-1), 101.6(Cl3-1), ll7.4( Ca or B-allyl), ll7.6( Cu or B-allyl), 169.4(Crcarbonyl). 58 Methyl 2,3-di-O-benzoyl-4,6-O-benzylidene-a-D-glucopyranoside (27) To a suspension of methyl-a-D—glucopyranoside ( 29g, 0.15 mole ) in dry DMF ( 60 ml ) were added a,0t—dimethoxytoluene ( 30g, 0.20 mole ) and dry p-TsOH ( 200 mg ) at room temperature. After stirrin g at room temperature for 18 hours, MeOH produced in the reaction mixture was removed by evaporation at reduced pressure. Anhydrous potassium carbonate ( 0.5 g) was added to neutralize the reaction mixture. After filtering the precipitate from the reaction mixture, the filtrate was concentrated by vacuum distillation at 40°C. Ether ( 20 ml) was poured into the residue and the reaction mixture was stood in the refrigerator ( 0°C ) over night. The product was precipitated further by adding pet. ether ( 200 ml ) to the ethereal solution. The precipitate was collected by filtration, dried in vacuo and weighed. The yield was 35g. To a solution of the product ( 35g, 0.12 mole ) in dry pyridine ( 120 ml) at 0°C was added benzoyl chloride (37.8 g, 0.26 mole ). The mixture was kept at 0°C for 0.5 hour. It was then stirred at room temperature for 4 hours and then poured into a mixture of distilled water ( 200 ml ) and ether ( 300 ml ). After stirring for additional 0.5 hour, the organic layer was separated and concentrated under reduced pressure. The residue was precipitated by a mixture of pet. ether ( 100 ml ) and ether ( 50 ml ). The precipitate was collected by filtration and the filter cake was dried in vacuo and weighed ( 33.4g ). Additional product ( 6.7g ) was isolated from the filtrate by flash column chromatography. Yield ; 40.1 g ( 54% ). TLC ; R1: = 0.50 ( CH2C12-ether : 20—1 ). 1H-NMR ( 300 MHz, CDCl3 ) 8 3.42(3H, S, OCH3), 3.85(1H, dd, J = 10 and 10 Hz, H-6(ax)), 3.90(1H, dd, J = 10 and 10 Hz, H-4), 4.08(1H, ddd, J = 10, 10 and 5 Hz, H-5), 4.37(1H, dd, J = 10 and 5 Hz, H-6(eq)), 5.17(1H, d, J = 3 Hz, H-l), 5.25(1H, dd, J = 10 and 3 Hz, H—2), 5.57(1H, s, H-benzyl), 6.07(1H, dd, J = 10 and 10 Hz, H-3), 7.29-8.01(15H, m, H-aromatic). 13C-NMR ( 75.5 MHz, CDC13) 8 97.8(C-l), 101.6(C-benzyl), 165.5(C-carbonyl), 166.0(C-carbonyl). 59 Methyl 2,3-di-O-benzoyl-6-O-benzyl-a-D-glucopyranoside (28) To a room temperature solution of 27 ( 33.4g, 67.6 mmole ) and borane-trimethylamine complex ( 14.8g, 203 mmole ) in dry toluene ( 1.2 l ) was added, dropwise anhydrous p-TsOH ( 35 g, 203 mmole ) at room temperature over 0.5 hour. After stirring at room temperature for 18 hours, the reaction mixture was concentrated under reduced pressure. Ether ( 300 ml ) and pet. ether ( 300 ml ) were added to the residue and the precipitate formed was filtered off. The filtrate was concentrated and the product was purified by flash column chromatography (pet. ether-ether : 1-2 ). Yield ; 30g ( 90% ). TLC ; R}: = 0.45 (pet. ether-ether : 20-1 ). 1H-NMR ( 300 MHz, CDC13 ) 8 3.41(3H, s, OCH3), 3.78-3.88(2H, m, H-6), 3.95(1H, m, H-5), 3.98(1H, dd, J = 10 and 10 Hz, H-4), 4.60(1H, d, J = 12 Hz, H-benzyl), 4.67(1H, d, J = 12 Hz, H-benzyl), 5.12(1H, d, J = 3 Hz, H—l), 5.25(1H, dd, J = 10 and 3 Hz, H-2), 5.75(1H, dd, J = 10 and 10 Hz, H-3), 7.27-7.98(15H, m, H-aromatic). Methyl 2,3-di-O-benzoyl-4-O-methyl-0t-D-glucopyranoside (29) To a solution of 28 (30.0g, 60.5 mmole ) in dry DMF ( 30 ml ) were added AggO ( 20.0g, 86.3 mmole ), anhydrous Ca2SO4 ( 20g ) and iodomethane ( 68.4g, 0.48 mole ) at room temperature. After stirring at room temperature for a further 18 hours, the reaction mixture was poured into ether ( 300 ml ). The precipitate was removed by filtering through celite and the filtrate was concentrated under reduced pressure and the residual DMF was removed by vacuum distillation. The product was further purified by passage through a silica gel column with 1 liter of a mixture of pet. ether-ether ( 2-1 ). After concentrating the filtrate, ethyl alcohol ( 120 ml ) and 10% palladium on activated carbon ( 10g ) were added. The reaction mixture was hydrogenated by attaching a hydrogen balloon to the reaction flask. After stirring the reaction mixture for 2 days, the reaction was checked for completion by TLC. The 6O precipitate in the reaction mixture was removed by filtration through celite and the filtrate was concentrated The product was purified by flash column chromatography. Yield ; 18.0g ( 71% ). TLC ; RF = 0.23 (pet. ether-ether : l-2 ). 1H-NMR ( 300 MHz, CDC13 ) 8 3.38(3H, s, OCH3), 3.45(3H, s, OCH3), 3.64(1H, dd, J = 10 and 10 Hz, H-4), 3.77-3.94(3H, m, H-5 and H-6), 5.06(1H, dd, J = 10 and 3 Hz, H-2), 5.10(1H, d, J = 3 Hz, H-l), 5.94(1H, dd, J = 10 and 10 Hz, H-3), 7.32-8.03(10H, m, H—aromatic). 13C-NMR ( 75.5 MHz, CDC13) 8 97.0(C-1), 165.6(C-carbonyl), 166.0(C-carbonyl). The alternative way for the preparation of 29 from 43 in Scheme 12 was the same as that employed for the preparation of 26 ( Yield; 65% ). l-O-Methyl-2,3-di-O-benzoyl-4-O-methyl-0t-D-glucuronic acid methylester (3 0) To a room temperature solution of 29 ( 17g, 40.5 mmole ) in dry DMF ( 90 ml ) was added pyridinium dichromate ( 91.4g, 0.24 mole ). The reaction mixture was stirred at room temperature for 18 hours after which it was treated with distilled water ( 800 ml ) and ethyl acetate ( 500 ml) and stirred for 0.5 hour. The organic layer was isolated and washed with a aqueous 4% HCl solution ( 300 ml ). The ethyl acetate layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, followed by further concentration under high vacuum. To the residue were added MeOH ( 600 ml ), anhydrous magnesium sulfate ( 20g) and p-TsOH ( 1.0g ). The reaction mixture was stirred at room temperature for 18 hours after which it was treated with sodium bicarbonate ( 1.5 g ) for 0.5 hour and concentrated under reduced pressure. The residue was treated with dichloromethane ( 400 ml ) for 0.5 hour and the precipitate was removed by filtration. The filtrate was concentrated and the residue was purified by flash column chromatography. Yield ; 12.0g ( 66% ). TLC ; R1: = 0.55 ( CHzClz-ether : 20-1 ). 61 1H-NMR ( 300 MHz, CDC13 ) 8 3.34(3H, s, OCH3), 3.36(3H, s, OCH3), 3.77(3H, s, OCH3), 3.79(1H, dd, J = 10 and 10 Hz, H—4), 4.23(1H, d, J = 10 Hz, H-5), 5.07(1H, dd, J = 10 and 3 Hz, H—2), 5.11(1H, d, J = 3 Hz, H-l), 5.87(1H, dd, J = 10 and 10 Hz, H—3), 7.25-7.95(10H, m, Hraromatic). 13C-NMR ( 75.5 MHz, CDC13) 8 97.4(C—1), 165 .4(C-carbonyl), 165 . 8(C-carbonyl), 169.4(C—carbonyl). Benzyl-a,B-D-glucopyranoside (32, a/B=l/l) To a suspension of B-D—glucose ( 60g, 0.33 mole) in benzyl alcohol ( 500 ml, 4.83 mole) and benzene ( 150 ml ) was added acetyl chloride ( 2.0 ml ). The reaction mixture was refluxed with a Dean-Stark apparatus for 18 hours to remove the water that was formed. After checking for completion of the reaction by 1H—NMR spectroscopy, the reaction mixture was treated with anhydrous potassium carbonate (4.0g ) at room temperature for 1 hour. Benzene in the reaction mixture was evaporated at reduced pressure, then the reaction flask was allowed to stand until all of the precipitate settled. The supernatant was carefully decanted into ether (200 ml ). This ethereal solution was stirred with distilled water ( 500 ml ) for 1 hour and the aqueous layer was isolated. The aqueous layer was washed again with ether ( 150 ml ) and then concentrated under reduced pressure and dried further under high vacuum. The residue was used for the next reaction without further purification. Yield ; 69g ( 77% ). 1H-NMR ( 300 MHz, D20 ) 8 3.10-3.78(4H, m, H(B)—2, H-3, H—4, H-5 and H—6), 3.39(1H, dd, J = 9 and 3 Hz, H(0t)-2), 4.34(1H, d, J = 9 Hz, H(B)—1), 4.42—4.78(2H, m, H=benzyl), 4.85(1H, d, J = 3 Hz, H(0L)-l), 7.20-7.31(5H, m, H-aromatic). 62 Benzyl 4,6-0-(4-methoxy)benzylidene-a,B-D-glucopyranoside (33, 0t/ [3:1/ 1) The procedure for the preparation of 33 was the same as that employed for the preparation of 42. Yield ; 37.7g ( 38% ). TLC; R1: = 0.36 ( CH2C12-MeOH : 20-1 ). 1H-NMR ( 300 MHz, DMSO—d6 ) 8 3.15-4.20(6H, m, H-2, H-3, H-4, H-5 and H-6), 3.73(3H, s, OCH3), 4.46(1H, d, J = 9 Hz, H(B)-1), 4.50(1H, d, J = 12 Hz, H(0t)-benzyl), 4.58(1H, d, J = 12 Hz, H(B)-benzyl), 4.70(1H, d, J = 12 Hz, H(0t)-benzyl), 4.78(1H, d, J = 12 Hz, H(B)-benzyl), 4.85(1H, d, J = 3 Hz, H(0t)-1), 5.50(1H, s, H(0t)—benzylidene), 5.51(1H, s, H(B)-benzylidene), 6.89-7.4l(9H, m, H-aromatic). Benzyl 2,3-di-O-benzyl-4,6-O-(p-methoxy)benzylidene-oz,B-D- gluc0pyranoside (34, OL/B=1/1) To a room temperature solution of 33 ( 16g, 41.2 mmole ) in dry THF ( 200 ml ) was added ( 3.3g, 82.5 mmole ) of a 60% dispersion of NaH in mineral oil and the mixture kept stirring at room temperature for 0.5 hour. Benzyl bromide ( 14. lg, 82.5 mmole ) was then added and stirring maintained for an additional 18 hours after which the reaction mixture was concentrated and the residue was treated with distilled water ( 100 ml ) and dichloromethane ( 200 ml ). The organic layer was isolated and dried over anhydrous magnesium sulfate. The drying agent was removed by filtration and the filtrate was concentrated and the concentrate was used for the next reaction without further purification. Yield; 22.3g ( 95% ). TLC; R}: = 0.30 ( CH2C12 ). 1H-NMR of a-isomer ( 300 MHz, CDC13 ) 8 3.53(1H, dd, J = 10 and 4 Hz, H-2), 3.58(1H, dd, J = 10 and 10 Hz, H-4), 3.66(1H, dd, J = 10 and 10 Hz, H-6), 3.80(3H, s, OCH3), 3.88(1H, ddd, J = 10, 10 and 5 Hz, H-S), 4.09(1H, dd, J = 10 and 10 Hz, 63 H(0t)-3), 4.17(1H, dd, J = 10 and 5 Hz, H—6), 4.55—4.93(6H, m, H-benzyl), 4.80(1H, d, J = 4 Hz, H-l), 5.50(1H, s, H-benzylidene), 6.89(19H, m, H-aromatic). Benzyl 2,3-di-O-benzyl-0t,B-D-glucopyranoside (35, a/B=1/l) Compound 34 ( 22.3g, 39.3 mmole ) was stirred with a mixture of acetic acid ( 100 ml ) and distilled water ( 50 ml ) at room temperature for 6 hours. The reaction mixture and additional distilled water ( 200 ml ) were transfered to a 1 liter beaker, and sodium bicarbonate was added to the beaker until no further gas evolution. The product was extracted twice with dichloromethane ( 200 ml ) from the aqueous solution. The combined organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and the product was purified by flash column chromatography. Yield; 17.6g ( 84% ). TLC; R}: = 0.32 (CH2C12—MeOH : 20-1 ). 1H-NMR of (rt-isomer ( 300 MHz, CDC13 ) 8 3.48(1H, dd, J = 10 and 4 Hz, H—2), 3.55(1H, dd, J = 10 and 10 Hz, H-4), 3.68(1H, dd, J = 10 and 3 Hz, H-5), 3.72(2H, d, J = 3 Hz, H-6), 3.87(1H, dd, J = 10 and 10 Hz, H-3), 4.50-5.05(6H, m, H-benzyl), 7.28-7.42(15H, m, H-aromatic). 13C-NMR of (it-isomer ( 75.5 MHz, CDC13) 8 62.2, 69.1, 70.3, 71.0, 72.7, 75.3, 79.7, 81.3, 95.5, 127.8-128.5(C-aromatic), 137.0, 137.9, 138.7. Benzyl 2,3-di-O-benzyl-6-0-triphenylmethyi-a,B-D-glucopyranoside (36, a/B: 1/1) To a solution of 35 ( 13.6g, 30.2 mmole ) in dry pyridine ( 50 ml ) was added triphenylmethyl chloride ( 9.3g, 33.2 mmole ). The reaction mixture was stirred at 70°C for 18 hours. When the reaction was complete, the reaction mixture was treated with dichloromethane ( 200 ml ) and distilled water ( 150 ml ). The organic layer was isolated and dried over anhydrous magnesium sulfate. After filtering, the filtrate was concentrated 64 under reduced pressure until most of the pyridine was removed The residue was subjected to flash column chromatography. Yield; 19.7g ( 94% ). TLC; R1: = 0.28 ( CH2C12 ). 1H-NMR of a—isomer ( 300 MHz, CDC13 ) 8 3.19(1H, dd, J = 12 and 6 Hz, H-6), 3.24(1H, dd, J = 12 and 4 Hz, H-6), 3.46(1H, dd, J = 10 and 4 Hz, H—2), 3.50(1H, dd, J = 10 and 10 Hz, H—4),3.69-3.76(1H, m, H-5), 3.77(1H, dd, J = 10 and 10 Hz, H-3), 4.81(1H, d, J = 4 Hz, H—l), 7.12-7.42(30H, m, H-aromatic). 13CNMR ( 75.5 MHz, CDC13) 8 63.7, 68.7, 70.3, 71.4, 72.7, 75.6, 79.6, 81.7, 86.7, 94.9, 127.0-128.7(C-aromatic), 137.1, 138.1, 138.8, 143.9. Benzyl 2,3-di-O-benzyl-4-O-methyI-0t,B-D-glucopyranoside (37, a/B: l/l) To a room temperature solution of 36 ( 27.5 g, 40.0 mmole ) in dry THF ( 180 ml ) was added 60% NaH in mineral oil ( 1.75g, 44.0 mmole ). After stirring the suspension at room temperature for 1 hour, iodomethane ( 9.4g, 66.3 mmole ) was added at room temperature over a period of 0.5 hour. The reaction mixture was stirred at room temperature for a further 18 hours and then concentrated. Dichloromethane (300 ml ) and distilled water ( 100 ml ) were added to the residue, and the organic layer was isolated and passed through a short column of silica gel. The filtrate was concentrated and the residue was treated with a mixture of 4% aqueous HCl solution (20 ml), acetic acid (40 ml) and dichloromethane ( 100 ml ) for 18 hours. The reaction mixture was then poured slowly into 20% K2CO3 aqueous solution (250 ml) with vigorous stirring. The organic layer was isolated and dried over anhydrous magnesium sulfate. After filtation, the filtrate was concentrated and the product was isolated by flash column chromatography. Yield; 12.0g ( 65% ). TLC; RF = 0.50 ( CH2C12-MeOH : 20-1 ). lH--NMR of a-isomer ( 300 MHz, CDC13 ) 8 3.18(1H, dd, J = 10 and 10 Hz, H-4), 3.37(1H, dd, J = 10 and 4 Hz, H-2), 3.49(3H, s, OCH3), 3.55(1H, m, H—5), 3.61(1H, 65 dd, J = 12 and 4 Hz, H-6), 3.68(1H, dd, J = 12 and 3 Hz, H-6), 3.88(1H, dd, J = 10 and 10 HZ, H-3), 4.45-4.91(6H, m, H-benzyl), 4.72(1H, d, J = 4 Hz, H-l), 7.19-7.35(15H, m, H-aromatic). 13C-NMR of a-isomer ( 75.5 MHz, CDC13) 8 60.8, 61.9, 69.2, 71.0, 73.1, 75.6, 79.7, 81.8, 95.6, 127.6-128.4(C-aromatic), 137.1, 138.1, 138.8. 1,2,3-Tri-O-benzyl-4-O-methyl-0t,B-D-xylo-hex-6-al (38, a/B=1/1) To the solution of acetic anhydride ( 90 ml) in anhydrous DMSO ( 1600 ml) was added 37 ( 9.0g, 19.4 mmole ). The reaction mixture was stirred at 30°C for 48 hours and then concentrated by vacuum distillation below 30°C. The product was isolated from the oily residue by flash column chromatography. Yield; 4.8g ( 54% ). TLC ; R1: = 0.23 (pet. ether-ether: 3-2 ). 1H-NMR ( 300 MHz, CDC13 ) 8 3.28(1H, dd, J = 10 and 9 Hz, H(0t)-4), 3.40-3.51(2H, m, H-2 and H(B)-4), 3.52(3H, s, OCH3), 3.54(3H, s, OCH3), 3.59(1H, d, J = 6 Hz, H(B)-5), 3.63(1H, dd, J = 9 and 9 Hz, H(0t)—3), 3.78(1H, dd, J = 10 and 1 Hz, H(0t)-5), 4.03(1H, dd, J = 9 and 9 Hz, H(B)-3), 4.11(1H, d, J = 10 Hz, H([3)-1), 4.50-4.99(7H, m, H—benzyl and H(0t)-1), 7.25-7.40(15H, m, H-aromatic), 9.67(1H, s, H(B)-aldehyde), 9.72(1H, d, J = 1 Hz, H(0t)-aldehyde). (l-Decyl)triphenylphosphonium iodide (39) To the solution of triphenylphosphine ( 3.15 g, 12 mmole ) in dry THF ( 10 ml ) was added 1-iododecane ( 2.7g, 10 mole ). This reaction mixture was stirred at 70°C for 18 hours. When the reaction was complete, the reaction mixture was concentrated under reduced pressure and the residue was treated with ether ( 30 ml ). The ethereal solution was stood in the refrigerator ( -20°C ) for 18 hours. The resultant precipitate was collected by filtration and dried in vacuo at 30°C. 66 Yield ; 5.0g ( 94% ). IH—NMR ( 300 MHz, CDC13 ) 8 0.80(3H, t, J = 8 Hz, CH3), 1.12-1.22(16H, m, CH2), 1.57(2H, m, CH2), 3.57(2H, m, PCHz), 7.65—7.82(15H, m, H—aromatic). 13C-NMR ( 75.5 MHz, CDC13) 8 13.9, 22.1, 22.2, 23.2, 29.0. 29.2, 30.1, 30.3, 31.5, 117.7(d, J = 83 Hz, PCH2), 128 .0- 1 35 .O(C—aromatic). Benzyl 2,3-di-O-benzyl-4-O-methyl-5-trans(1-undecene-l-yl)-0t,[3-D- xylopyranoside (40, a/B=l/l) To the solution of phosphonium iodide 39 ( 390 mg, 0.75 mmole ) in dry THF ( 12 ml ) was added 2.06M n—BuLi in hexane (0.37 ml, 0.76 mmole ) at -78°C. This solution was stirred at 0°C for 1 hour and cooled again to —78°C. Intermediate 38 ( 160 mg, 0.35 mole ) dissolved in dry THF ( 3 ml ) was then added to the reaction mixture, which was then warmed up to room temperature and stirred for 18 hours after which time it was complete. The reaction mixture was concentrated under reduced pressure and the residue was dissolved in dichloromethane ( 20 ml ). The dichloromethane solution was washed with distilled water ( 10 ml ) and dried over anhydrous magnesium sulfate. After filtration, the filtrate was concentrated and the product was purified by flash column chromatography. Yield ; 150 mg ( 74% ). TLC ; RF = 0.37(0t), 043(8) ( CH2C12 ). 1H-NMR of (it-isomer ( 300 MHz, CDC13 ) 8 0.85(3H, t, J = 7 Hz, CH3), 1.23-1.40(14H, m, CH2), 2.12(2H, m, CH2), 3.01(1H, dd, J = 10 and 10 Hz, H-4), 3.48( 1H, dd, J = 10 and 3 Hz, H-2), 3.49(3H, s, OCH3), 3.95( 1H, dd, J = 10 and 10 Hz, H-3), 4.43( 1H, dd, J = 10 and 10 Hz, H-5), 4.51-4.95(6H, m, H-benzyl), 4.77(1H, d, J = 3 Hz, H-l), 5.35( 1H, ddt, J = 10, 10 and le, H-6(vinyl)), 5.71( 1H, dt, J = 10 and 7 Hz, H-7(vinyl)), 7.25-7.41(15H, m, H-aromatic). 1H-NMR of B-isomer ( 300 MHz, CDC13 ) 8 0.85(3H, t, J = 7 Hz, CH3), 1.23-1.40(14H, m, CH2), 2.15(2H, m, CH2), 3.08(1H, dd, J = 10 and 10 Hz, H-4), 'Ir 67 3.45( 1H, dd, J = 10 and 10 Hz, H—2 or H—3), 3.49(3H, s, OCH3), 3.53( 1H, dd, J = 10 and 10 Hz, H-2 or H-3), 4.00( 1H, dd, J = 10 and 10 Hz, H-5), 4.52(1H, d, J = 10 Hz, H-l), 4.57-4.96(6H, m, H-benzyl), 5.44( 1H, ddt, J = 10, 10 and 1H2, H—6(vinyl)), 5.73( 1H, dt, J = 10 and 8 Hz, H-7(vinyl)), 7.25-7.40(15H, m, H-aromatic). 13C-NMR of Ot-isomer ( 75.5 MHz, CDC13) 8 14.0, 22.5, 28.3, 29.5, 29.7, 32.0, 61.0, 66.5, 68.7, 73.2, 75.7, 79.5, 81.6, 84.6, 95.2, 126.5-139.0(C-aromatic and C-vinyl). 4-O-methyl-6-undecyl-0t,B-D-xylose (41, a/lell) To the solution of 40 (120 mg, 0.20 mole ) in MeOH ( 5 ml ) was added 10% palladium in activated charcoal (250 mg, 0.24 mole). This suspension was stirred under hydrogen using hydrogen filled balloon attached to the reaction flask, at room temperature for 18 hours. When the reaction was complete, the reaction mixture was filtered through celite and the filtrate was concentrated under reduced pressure. The residue was used for the subsequent reaction without further purification. Yield; 66 mg ( 100% ). TLC; R1: = 0.35 ( CHgClz-MeOH: 10-1 ). lH-NMR ( 300 MHz, CDC13 ) 8 0.88(3H, t, J = 6 Hz, CH3), 1.25-1.80(20H, m, CH2), 2.83(1H, dd, J = 9 and 9 Hz, H(a or B)-4), 2.89(1H, dd, J = 9 and 9 Hz, H(a or |3)-4), 3.21(1H, m, H(B)-5), 3.32(1H, dd, J = 9 and 9 Hz, H(B)-2), 3.50-3.62(2H, m, H(oc)—2 and H(b)-3), 3.58(3H, s, OCH3), 3.72(1H, m, H(a)—5), 3.81(1H, dd, J = 9 and 9 Hz, H(oc)-3), 4.52(1H, d, J = 9 Hz, H(B)—1), 5.20(1H, d, J = 3 Hz, H(a)-1). (R)-3-Hydroxytetradecanoic acid (8) To the suspension of triol 41 ( 66 mg, 0.21 mmole ) in 0.1 M aqueous NaOH (4 ml, 0.4 mmole )solution was added 30% aqueous H202 solution (45 mg, 0.4 mmole ). The reaction mixture was then stirred at 80°C for 48 hours. After cooling to room temperature, dichloromethane ( 10 ml ) was added to the reaction mixture. 4% aqueous HCl solution was added to the bilayer mixture with vigorous stirring until the pH 68 of the aqueous layer was strongly acidic ( pH < 2 on pH-paper ). The organic layer was then isolated and concentrated. The product in the concentrate was identified by 1H-NMR analysis. Yield; 60% (based on 1H-NMR analysis ). 1H-NMR ( 300 MHz, CDC13 ) 8 0.87(3H, t, J = 7 Hz, CH3), 1.20-1.60(20H, m, CH2), 2.47(1H, dd, J = 17 and 9 Hz, COCH2), 2.58(1H, dd, J = 17 and 3 Hz, COCHz), 4.04(1H, m, CHOH). Methyl 4,6-0-(4-methoxy)benzylidene-oc-D-glucopyranoside (42) To a suspension of methyl-or—D-glucopyranose ( 60g, 0.31 mole ) in dry DMF ( 100 ml ) were added p—methoxybenzaldehyde ( 44.4g, 0.33 mole) and anhydrous p-TsOH ( 0.5 g ). After stirring at room temperature for 2 hours, toluene ( 200 ml ) was added to the reaction mixture and evaporated at reduced pressure. This evaporation step with toluene was repeated three times. Anhydrous potassium carbonate ( 1.5g ) was added to the reaction mixture which was subsequently concentrated by vacuum distillation. The residue was treated with a mixture ( 500 ml ) of pet. ether and ether ( 1 : 1 ), and the precipitate formed was collected by filtration. The product was extracted from the filter cake with 1.2 liter of 5% methanolic dichloromethane. The extract was passed through a short silica gel ( 20g ) column and concentrated Yield; 46g ( 48% ). TLC; R1: = 0.30 ( CHZClz-MeOH : 20-1 ). 1H-NMR ( 300 MHz, DMSO-d6 ) 8 3.30(3H, s, OCH3), 3.30-3.37(2H, m, H-3 and H-6), 3.52-3.60(2H, m, H-2 and H-5), 3.65(1H, dd, J = 10 and 10 Hz, H-4), 3.74(3H, s, OCH3), 4.12(1H, dd, J = 10 and 4 Hz, H-6), 5.13(1H, d, J = 4 Hz, H-l), 5.49(1H, s, H-benzyl), 6.90( 2H, d, J = 10 Hz, H—aromatic), 7.35(2H, d, J = 10 Hz, H-aromatic). 69 Methyl 2,3-di-0-benzoyl-6-O-triphenylmethyl-a-D-glucopyranoside (43) The procedure for the preparation of 43 was the same as that employed for the preparation of 25 from 23. Yield; 57%. TLC ; R}: = 0.27 ( CH2C12-pet. ether : 2-1 ). lH-NMR ( 300 MHz, CDC13 ) 8 3.42(3H, s, OCH3), 3.43-3.54(2H, m, H-4 and H-5), 3.85-3.94(2H, m, H-6), 5.13(1H, d, J = 3 Hz, H-l), 5.23(1H, dd, J = 10 and 3 Hz, H—2), 5.75(1H, dd, J = 10 and 10 Hz, H-3), 7.21-7.99(35H, m, H-aromatic). 13C—NMR ( 75.5 MHz, CDC13) 8 87.2(C-uityl), 96.9(C-1), 166.0(C-carbonyl), 167.0(C-carbonyl). 2,3-Di-O-benzoyl-1,4-di-O-methyl-6-bromo-6-deoxy-0t-D- glucopyranoside (44) To a solution of 29 ( 6.8g, 16.3 mmole ) in dry THF ( 50 m1 ) was added tetrabromomethane ( 10.84g, 32.7 mmole ) and triphenylphosphine ( 8.6g, 32.8 mmole ). After stirring at room temperature for 16 hours, THF was evaporated under reduced pressure. The residue was treated with ether ( 250 ml ) and the precipitate formed was filtered through celite. The filtrate was concentrated and the product was purified by flash column chromatography. Yield ; 6.7g ( 86% ). TLC; RF = 0.45 ( CH2C12 ). 1H-NMR ( 300 MHz, CDC13 ) 8 3.41(3H, s, OCH3), 3.48(3H, s, OCH3), 3.58(1H, dd, J = 10 and 10 Hz, H-4), 3.68-3.77(2H, m, H-6), 3.91-3.98(1H, m, H-S), 5.11(1H, dd, J = 10 and 3 Hz, H-2), 5.13(1H, d, J = 3 Hz, H-l), 5.94(1H, m, H-3), 7.32—8.02(10H, m, H-aromatic). l3C—NMR ( 75.5 MHz, CDC13) 8 79.8(C-6), 97.0(C-1), 165 .6(C-carbonyl), 165 .9(C—carbonyl). 7O 2,3-Di-O-benzoyl-6-de0xy-1,4-di-O-methyl-6-triphenylphosphino-a-D- glucopyranosyl bromide (45) Compound 44 ( 6.8g, 14.2 mmole ) and triphenylphosphine ( 3.72g, 14.2 mmole ) were dissolved in dry DMF ( 30 ml ). The solution was then stirred at 100°C for 2 days. After concentrating the reaction mixture by vacuum distillation, the residue was purified by flash column chromatography. Yield ; 2.5g ( 24% ). TLC; RF = 0.27 ( CH2C12-MeOH : 10-1 ). 1H-NMR ( 300 MHz, CDC13 ) 8 3.53(3H, s, OCH3), 3.62(3H, s, OCH3), 3.71(1H, ddd, J = 18, 18 and 2 Hz, H—6), 4.00(1H, ddd, J = 10, 10 and 2 Hz, H-5), 4.12(1H, dd, J = 10 and 10 Hz, H-4), 4.86(1H, d, J = 3 Hz, H—l), 5.03(1H, dd, J = 10 and 3 Hz, H—2), 5.37(1H, ddd, J = 18, 10 and 10 Hz, H-6), 5.73(1H, dd, J = 10 and 10 Hz, H—3), 7.20—8.00(25H, m, H-aromatic). 13C-NMR ( 75.5 MHz, CDC13) 8 97.4(C-1), 118.3(d, J = 87 Hz, C-6), 162.3(C-carbonyl), 165.5(C-carbonyl). 1,2,3-Tri-O-benzyl-6-bromo-6-deoxy-4-O-methyl-0t,B-D-glucopyranoside (46, a/le/l) The procedure for the preparation of 46 was the same as that employed for the preparation of 44. Yield; 71%. TLC; RF = 0.38 ( CH2C12 ). 1H-NMR of oc—isomer ( 300 MHz, CDC13 ) 8 3.15(1H, dd, J = 10 and 10 Hz, H-4), 3.42(1H, dd, J = 10 and 4 Hz, H—2), 3.49(2H, d, J = 4 Hz, H-6), 3.52(3H, s, OCH3), 3.68(1H, dt, J = 10 and 4 Hz, H-5), 3.88(1H, dd, J = 10 and 10 Hz, H-3), 4.44-4.92(6H, m, H-benzyl), 4.76(1H, d, J = 4 H22, H-l), 7.20-7.36(15H, m, H-aromatic). 13C—NMR of (it-isomer ( 75.5 MHz, CDC13) 8 33.5, 61.0, 69.1, 69.8, 73.0, 75.6, 79.8, 81.5, 81.6, 95.5, 127.5-138.7(C—aromatic). 71 N,N’-bis-tetradecyl-(2R,3R)-tartaric acid diamide (10) To a solution of L-tartaric acid ( 7.0g, 46.6 mmole ) in methanol ( 50 ml ) were added p—TsOH ( 0.2g) and anhydrous magnesium sulfate ( 5 g ). After stirring at room temperature for 18 hours, sodium bicarbonate ( 1.0g ) was added to the reaction mixture. All precipitate was filtered away using celite and the filtrate was concentrated under reduced pressure. The residue was redissolved and passed through a short silica gel column ( CH2C12—MeOH : 20-1 ), the suspension was concentrated and the concentrate was treated with l—tetradecylamine (24g, 0.11 mole) and MeOH ( 150 ml ) at 70°C for 20 hours. After cooling the reaction mixture to room temperature, the precipitated product was collected by filtration. The filter cake was washed twice with MeOH ( 50 ml ) and dried in vacuo. Yield; 25.3g (97% ). TLC; R1: = 0.32 ( CH2C12-MeOH : 20-1 ). (For NMR analysis, the product was acetylated in acetic anhydride and pyridine due to its poor solubility in the usual NMR solvents. The diacetate of the product was then analyzed by NMR spectroscopy.) 1H-NMR ( 300 MHz, CDC13 ) 8 0.85(6H, t, J = 7 Hz, CH3), l.20-1.48(48H, m, CH2), 2.13(6H, s, CH3), 3.10-3.40(4H, m, NCH2), 5.56(2H, s, CH). 13C-NMR ( 75.5 MHz, CDC13) 8 39.8(NHCH2), 72.2(CH), 166.1(C—carbonyl), 169.5(C-carbonyl). N,N’-bis-tetradecyl-2,3-O-isopropylidene-(2R,3R)-tartaric acid diamide <4 7) To a suspension of 10 ( 16g, 28.6 mmole ) in 2,2-dimethoxypropane ( 150 ml ) was added p-TsOH ( 0.2g ). After stirring at room temperature for 4 hours, 50 ml of the solvent was removed from the reaction mixture by fractional distillation, and the residual solution was cooled to room temperature and treated with anhydrous potassium carbonate ( 1.0g ). After evaporating all 2,2-dimethoxypropane from the reaction mixture, the residue was treated with distilled water ( 100 ml ) and the precipitate formed was collected 72 by filtration. The filter cake was dissolved in ether ( 200 ml ) and the resultant ethereal solution was filtered through celite to remove a small amount of precipitate in the solution. The filtrate was dried over anhydrous magnesium sulfate and concentrated The residue was used for the next reaction without further purification. Yield ; 17.6g ( 100% ). TLC; R}: = 0.50 ( CH2C12-MeOH : 40-1 ). 1H-NMR ( 300 MHz, CDC13 ) 8 0.85(6H, t, J = 6 Hz, CH3), 1.21-1.55(48H, m, CH2), 1.47(6H, s, C(CH3)2), 3.27(4H, td, J = 6 and 6 Hz, NCH2), 4.48(2H, s, CH). N,N’-bis-tetradecy|-l,4-diamino-(2S,3S)-dihydroxybutane iSOpropylidene acetal (48) ‘ To a solution of 47 (9.4g, 15.6 mmole ) in dry ether ( 50 ml ) was added lithium aluminium hydride ( 2.0g, 52.6 mmole ). After refluxing for 16 hours, ethylacetate ( 3 ml ) was added to the reaction mixture to quench the reaction. The reaction mixture was treated with additional ether (400 ml) and 20% aqueous NaOH solution (200 ml ) at room temperature for 20 hours. The organic layer was isolated and dried over anhydrous magnesium sulfate, filtered and the filtrate concentrated The concentrate was treated with pet. ether ( 200 ml ). The precipitate formed was removed by filtration and the filtrate was concentrated. The concentrate was subjected to flash column chromatography ( CH2C12-MeOH-(Et)2NH : 40-2-1 ). Yield ; 6.9g ( 77% ). TLC; R}: = 0.20 ( CH2C12-MeOH-(Et)2NH : 40-2-1 ). 1H-NMR ( 300 MHz, CDC13 )8 0.85(6H, t, J = 7 Hz, CH3), 1.20-1.50(48H, m, CH2), 1.36(6H, s, CH3), 2.61(4H, t, J = 6 Hz, NCHz), 2.73(2H, dd, J = 10 and 4 Hz, CH2N), 2.82(2H, dd, J = 10 and 6 Hz, CHZN), 3.82-3.90(2H, m, CH). 13C-NMR ( 75.5 MHz, CDC13) 5 50.0(CH2NH), 51.9( CHZNH ), 79.0(CH), 109.0(OCO). 73 N,N’-bis-tetradecyl-1,4-dimethylamino-(2S,3S)-dihydroxybutane isopropylidene acetal di-methylsulfate salt (TM3 2CH3SO4') To a solution of 48 ( 8.7g, 15.0 mmole ) in dichloromethane ( 40 ml ) were added dimethyl sulfate ( 7.2g, 57.1 mmole ), anhydrous potassium carbonate ( 9.46g, 68.6 mmole ) and TBAB ( 0.1g). After stirring at room temperature for 18 hours, the reaction mixture was treated with distilled water ( 100 ml) for 18 hours. The organic layer was isolated and dried over anhydrous magnesium sulfate. After filtration, the filtrate was concentrated and the residue was dried in vacuo. Yield; 4.2g ( 33% ). 1H-NMR ( 300 MHz, CDC13 ) 8 0.80(6H, t, J = 7 Hz, CH3), 1.15-1’.30(44H, m, CH2), 1.40(6H, s, CH3), 1.60-1.80(4H, m, CH2), 3.16(6H, s, NCH3), 3.18(6H, s, NCH3), 3.30-3.40(4H, m, NCHz), 3.60(6H, s, CH3SO4'), 3.77(2H, dd, J = 13 and 7 Hz, CHZN), 3.98(2H, dd, J = 13 and 1 Hz, CH2N), 4.42(2H, m, CH). 13C-NMR ( 75.5 MHz, CDC13) 8 51.5, 54.2, 58.5, 64.2, 66.0, 72.6(CH), 112.8(OCO). N ,N ’-bis-tetradecyl-2,3-O-methylene-(2R,3R)-tartaric acid diamide (49) To the suspension of 10 ( 1.32g, 2.35 mole ) in dichloromethane ( 50 ml) were added anhydrous potassium carbonate ( 2.2g, 16 mole ) and 18-crown-6 (0.27g, 1.0 mmole ). To this solution was added diiodomethane ( 1.5g, 5.6 mmole ) at room temperature. The reaction mixture was refluxed for 18 hours. When the reaction was complete, the reaction mixture was cooled to room temperature and treated with distilled water ( 50 ml ). The organic layer was isolated and dried over anhydrous magnesium sulfate. After filtration, the filtrate was concentrated and the residue was purified by flash column chromatography. Yield; 1.2g ( 89% ). TLC; R}: = 0.50 ( CH2C12-MeOH : 30-1 ). 74 1H-NMR ( 300 MHz, CDC13 ) 8 0.85(6H, t, J = 6 Hz, CH3), 1.20-1.55(48H, m, CH2), 3.26(4H,m, NCHZ), 4.44(2H, s, CH), 5.11(2H, s, OCHzO). 13C-NMR ( 75.5 MHz, CDC13) 8 77.5(CH), 96.1(OCH20), 169.0(C—carbonyl). N,N’-bis-tetradecyl-1,4-diamino-(2S,3S)-dihydroxybutane methylene acetal (50) The procedure for the preparation of 50 was the same as that employed for the preparation of 48. Yield ; 79%. TLC ; R}: = 0.20 ( CHZClg—MeOH-(EthNH : 40-2-1 ). 1H-NMR ( 300 MHz, CDC13 ) 8 0.85(6H, t, J = 6 Hz, CH3), 1.20-1.55(48H, m, CH2), 2.62(4H, t, J = 6 Hz, NCHZ), 2.75(2H, dd, J = 12 and 3 Hz, CHzN), 2.83(2H, dd, J = 12 and 8 Hz, CHgN), 3.82(2H, m, CH), 4.99(2H, s, OCH20). 13C—NMR ( 75.5 MHz, CDC13) 8 49.8(CH2NH), 51.0(CH2NH), 78.5(CH), 94.0(OCO). N,N’-bis-tetradecyl-1,4-dimethylamino-(ZS,3S)-dihydroxybutane methylene acetal di-methylsulfate salt (TM4 2CH3SO4') The procedure for the preparation of TM4 was the same as that employed for the preparation of TM3. Yield ; 45%. lH-NMR ( 300 MHz, CDC13 ) 8 0.85(6H, t, J = 7 Hz, CH3), 1.20—1.35(44H, m, CH2), 1.65-1.83(4H, m, CH2), 3.23(6H, s, NCH3), 3.25(6H, s, NCH3), 3.42(4H, t, J = 6 Hz, NCHZ), 3.68(6H, s, CH3SO4'), 3.85(2H, dd, J = 12 and 7 Hz, CH2N), 4.15(2H, dd, J = 12 and 1 Hz, CHQN), 4.49(2H, m, CH), 5.15(2H, s, OCH20). 13C-NMR ( 75.5 MHz, CDC13) 8 51.2, 51.3, 54.5, 64.0, 66.5, 73.0(CH), 112.8(OCO). 75 REFERENCES 1. T. J. Sayers, I. Macher, J. Chung and E. Kugler, J. Immun., 1987, 138, 2935. 2. O. Westphal and O. Luderitz, Angew. Chem, 1954, 66, 407. 3. M. Imoto, H. Yoshimura, N. Sakaguchi, S. Kusumoto and T. Shiba, Tetrahedron Lett., 1985, 26, 1545. 4. M. Imoto, S. Kusumoto, T. Shiba, H. Naoki, T. Iwashita, E. Th. Rietschel, H. W. Wollenweber, C. Galanos and O. 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Kyung-Il Kim and Rawle l. Hollingsworth, Tetrahedron Lett., 1994, 1031. 28. E. J. Corey and Greg Schmidt, Tetrahedron Lett., 1979, 399. 29. Rabindra N. Rej, John N. Glushka, Warren Chew and Arthur S. Perlin, Carbohydr. Res., 1989, 189, 135. ‘S‘JH" 4;. .,V V'R' ' ryv \ ”$3172; 1.. .1 1117. v; 3‘11") 11.1 "'v--1w»1:u . 11:11-1:, r .;- _ |rvt~ ‘ y-.‘.. . r ,~.__ 131;. 11., H "3’ . mm. 11:, "NW!" 11...!‘IK 1 1- m. ‘ 7'“ i: _: when} 1 51“.“... "'33 f: I'nlr‘; 1 ,. q 1 r- " ' 11 1 1 ,. ,.1 NWT ~- . 11‘ 1 I 1 1 ‘r111'1 4‘11. "11 rl’y‘ m»n11-1..,"r' l‘ ' 11n- 1 111 rm», ’Qf‘j‘; ' 'I‘1 r! ” 11."1—,'.K1‘1.V1" 'r_ 1.1.1,“. ,J 1» 1' V r “‘"v *3 mm '1’?ny 1r 111,,I 1 1‘ QN STQTE UNIV mil Hill ‘li iii HIO . .;. ‘:‘.“‘:' 1 1 ; 41 ' 111 r'v'111111',1 1.111.: -1‘11‘-1-h1:!.r:.:. LIBRARIES .1111. v.1. m 11:. "131““ ....J .1.‘ :1 1. J...