JP2000507246A - Solid phase lipoglycopeptide libraries, compositions and methods - Google Patents

Abstract

  (57) [Summary] The present invention relates to a library of unique substances and compositions having the general formulas R-P-S-L and P-S-L, representing solid support-bound lipoglycopeptide and free lipoglycopeptide, respectively. Combination methods for preparing various libraries are described, as well as assays for detecting biological activity on selected ones. In particular aspects, bacterial peptidoglycan synthesis inhibitors are described.

Description

DETAILED DESCRIPTION OF THE INVENTION           Solid phase lipoglycopeptide libraries, compositions and methods 1.Field of invention   The present invention relates to the field of combinatorial library synthesis and drug discovery. Identify And the present invention relates to substances and compositions which mimic inhibitors of bacterial peptidoglycan synthesis. Regarding synthesis. The substances and compositions of the present invention are useful in treating infectious diseases. It is useful as a product candidate, for example, an antibiotic. 2. Background of the Invention   Bacterial peptidoglycan biosynthesis occurs through a complex multi-step process. Biosynthesis Early in the process, the synthesis of lipid-binding disaccharide-peptide monomer units Is involved. These monomer units then activate transglycosylase activity. It binds along with the growing peptidoglycan chains by the indicated enzymes. Continued Pentapeptide at the end of the peptidoglycan chain has transpeptidase activity It is believed that the enzyme is cross-linked by an enzyme or a separating enzyme. Transpepti The dase reaction is a β-lactam antibiotic such as penicillins and cephalosporins. Inhibited by substances. For example, Matsuhashi, M., Bacterial Cell Wall, J .-M Ghuysen and R. Hakenbeck (eds.), Elsevier Science B. V. (1994) pp.55-71 See also.   The structure of a naturally occurring lipid-linked glycopeptide intermediate is known. In this The interstitial is composed of N-acetylmuramic acid, the reducing end of which is pyrophosphoundecapre L-Ala-D-Glu- (A_Twopm / Lys) -D-A la-D-Ala binds to the linked D-lactyl group. A_Twopm represents diaminopimelic acid . This intermediate can also be designated as UDP-MuNAc-pentapeptide. Then this Intermediates are glycosyl salts that convert N-acetylglycosamine to N-acetylmuramic acid By the transferase-mediated addition, the lipid-binding disaccharide-peptide Convert to Nomer Unit. For example, Bacterial Cell W of Van Heijenoort, J. all, J. -M Ghuysen and R. Hakenbeck (eds.), Elsevier Science B. V. (1994) See pp.39-54.   Some designed to inhibit bacterial cell wall transglycosylation There has been described a method for synthesizing the compound of the above. For example, Hecker, S.M. J. La ”Synthesis of Compounds Designed to Inhibit Bacterial Cell Wall Tra nsglycosylation "J. Org. Chem. (1990) 55: 4904-4911. However, no active compounds have been described. Hecker and others Thus, the proposed compounds did not show any significant activity.   Multidrug-resistant bacteria, specifically vancomycin-resistant Enterococcus and methicillus With the emergence of bacteria such as phosphorus-resistant Staphylococcus, new anti-infective drugs Searching repeatedly for has been uplifted. Currently used cells Most of the "wall" inhibitors are late in peptidoglycan biosynthesis, i.e., Inhibits transglycosylation or transpeptidation. I Therefore, most drug-resistant bacteria have acquired resistance to these antibiotics. Came.   Thus the disaccharide-peptide monomer unit encoded by the murG gene That inhibit N-acetylglucosaminyltransferase involved in the biosynthesis of If you can use it, you can be a candidate. Bacterial murG has been cloned Its DNA sequence has been determined. Recombinant murG gene products are also known. This gene May the N-terminal half of the product be responsible for the binding of lipid intermediates? Can be further inferred. However, the part of the enzyme that catalyzes the transferase activity Minutes have not been decided.   Flavo Phosphoripol from Grey-Green Streptomyces Group A known antibiotic produced that inhibits glycosyltransferase Thereby hinder the biosynthesis of peptidoglycan. This is mainly cattle Used as a formance enhancer, ie it can fatten cattle feed To increase sex. Flavophospholipol is a gram-positive and gram-negative bacterium Active against gram-positive bacteria, but usually It shows a fairly broad spectrum with such activity. However, Gram-positive bacteria also -Negative bacteria also carry resistance to extrachromosomally induced antibiotics. I'm sorry. In fact, penicillin, tetracycline, chloramphenicol and Bacteria emerge in mammals resistant to antibiotics such as erythromycin and erythromycin That Quite a lot. Therefore, against existing antibiotics such as flavophospholipol The evolution of resistant bacteria is virtually certain.   As a result, to design and discover new anti-infectives such as antibiotics, Constant, challenging, and significant efforts are made. Manufacture of drug or nutrition candidates And techniques to facilitate identification are always being considered. These demands are greatly met It never changes and keeps changing. 3. Summary of the Invention   Accordingly, the present invention provides for the preparation of a library of unique substances having a given general formula. By providing a combination of methods for synthesizing and discovering new anti-infectives Dan is, of course, trying to satisfy the need for such drugs. is there. Specifically, the present invention presents potential drug or nutrition candidates, respectively. Provides solid-phase approaches that allow the synthesis of numerous materials and compositions such as I do.   An important object of the present invention is to provide a solid phase containing a plurality of unique substances represented by the following chemical formula: To produce a lipoglycopeptide library. In the formula, the substituent R is a solid support, and the substituent P is one or more amino acids. , Peptide, or polypeptide, the substituent S is one or more sugars, and Substituent L comprises one or more lipids.   The object of the present invention is to provide the following chemical formula Solid-phase lipoglycopeptide library with multiple unique substances represented by The present invention provides a method for preparing a yeast cell, the method comprising the following steps. That is, (a) the following chemical formula:One or more substituents R, P and FG represented by₁Providing a In this formula, R is one or more amino acids, peptides, or polypeptides. A substituent P containing a peptide, wherein at least one of the substituents P participates in a bond forming reaction. First functional group FG that can participate₁A solid support that binds the substituent P with ( b) the following chemical formula: One or more substituents S, L and FG represented by_TwoProviding a In this formula, S is one or more sugars and at least one of the sugars Is a sugar that binds a substituent L containing one or more lipids, A second functional group FG in which at least one sugar of group S can participate in the bond formation reaction_TwoHas You. (c) First functional group FG₁One of the substituents P having_TwoA substituent S having One or more of them so that a bond-forming reaction to form a bond with The above substituents R, P and FG₁And one or more substituents S, L and FG_TwoConcatenate with This is the process of   Alternatives are also being planned here. In certain embodiments, the following formula: Solid-phase lipoglycopeptide library with multiple unique substances represented by Lie is prepared by the following steps. That is, (a) the following chemical formula: One or more substituents R, P and FG represented by₁Providing a In this formula, R is one or more amino acids, peptides, or polypeptides. Petit A substituent P containing at least one of the substituents Functional group FG that can participate in₁A solid support that binds the substituent P with (b) Next The chemical formula of One or more substituents S, FG represented by_TwoAnd FG_ThreeThe process of providing Wherein S is one or more sugars and at least one of its substituents S A second functional group, FG, in which one sugar can participate in the first bond formation reaction_TwoWith The same or another of its substituents S participates in the subsequent bond-forming reaction Third functional group FG that can be made_ThreeIt has. (c) first functional group FG₁One of the substituents P with 2-functional group FG_TwoThe first bond-forming reaction that forms a bond with the sugar of the substituent S having Proceed to provide one or more initial bond-forming reaction products , One or more substituents R, P and FG₁And one or more substituents S, FG_Two And FG_ThreeIs a step of combining (d) the following chemical formula: One or more substituents L and FG represented by_FourIn the process of providing Wherein the substituent L is one or more lipids, wherein at least one of the A fourth functional group, FG, in which at least one lipid can participate in the subsequent bond-forming reaction_FourEquipped ing. (e) Third functional group FG_ThreeOne of the substituents S with a fourth functional group FG_FourWith a substituent One to allow the subsequent bond formation reaction to form a bond with the L lipid Or more initial bond-forming reaction products and one or more substituents L and F G_FourIs a step of combining   Yet another object of the present invention is the following chemical formula: From the substance shown in the following, obtained by cleaving the substituent P from the substituent R Chemical formula, With respect to the selected composition represented by In the formula, the substituent R is a solid support, The substituent P can be one or more amino acids, peptides, or polypeptides, substituted The group S contains one or more sugars and the substituent L contains one or more lipids. No. Further, a synthetic composition represented by the chemical formula shown above, that is, P-S-L, particularly a combination Compositions synthesized by the sedimentation method are also being planned.   Other objects of the present invention will be apparent to those of skill in the art in light of the disclosure provided herein. It will be obvious. 4. Brief description of drawings   FIG. 1 shows the overall procedure for conjugating a sugar-containing unit to a solid support-bound peptide. Figure 2 shows a schematic for a physical combination approach. Cleaved from the solid support After that, each unique combination is performed in a particular well.   FIG. 2 shows that the substituent S of the sugar linking the lipid substituent L is linked to the solid support-bound peptide. Show the sequence of steps that make up the parallel synthesis approach when connecting I have. Following the conjugation step, the step obtained by removing the protecting group and the solid support A step of cleaving the lipoglycopeptide is performed. Alternatively, the substituents S and L May be continuously bound to a solid support-bound peptide.   FIG. 3 is a diagram that favors the progress of a condensation reaction involving the functional groups of the substituents of P, S and L. 2 shows structural formulas of various coupling reagents.   FIG. 4 shows the sugar substituent disaccharide used in certain library embodiments of the invention. Some examples of the basic building block structures are shown.   FIGS. 5, 5a and 5b illustrate lipid and peptide groups preferred in certain embodiments of the invention. Some examples of the foundation unit structure are shown.   Figure 6 shows the coupling, deprotection, and cleavage of specific R-P and S-L couples. FIG.   FIG. 7 shows the activity of the library produced by the combination approach of the present invention. Is a schematic diagram of a high throughput screening method designed to You. However, other assays are available. Glycotrans The probe may be constructed using the binding region of the active site of ferase. Followed Probes screen lipoglycopeptides with affinity for the enzyme Possible ligand is directly bound to the solid support for detection Even if it is, it can be used.   Figure 8 shows four coupling trials consisting of, for example, EEDQ, HATU, HBTU, or PyBOP. More preferred is to couple the S-L substituent to the P-R substituent using one of the drugs FIG. 4 is a schematic diagram showing steps of a method. The process of deprotection and cleavage is also shown. 5. Detailed description of preferred embodiments   As with the objectives of the present invention, this method provides a wide variety of potential anti-infectives, Combination apps for synthesizing nutrients or performance enhancers The roach is also considered. In certain aspects of the invention, a parallel synthesis method is illustrated. . However, the invention is equally applicable to other methods, such as the "mix and split" method. It is possible. In fact, the desirable constituents of the unique substance bound to the solid phase (immediately After the design and selection of the `` basic unit structure ''), the It can be easily evaluated by a dynamic style, in this case a particular parcel For example, a planar solid surface may be assigned to a particular combination of molecular components.   When screening for specific libraries, use There are so many opportunities to discover enhancer ingredients, or nutrients Will.   An important object of the present invention is represented by the chemical formula as defined above, i.e., R-P-S-L. To produce solid-phase lipoglycopeptide libraries containing unique substances is there. The library of the present invention contains one or more copies of each unique substance. May be present. This allows a large number of screening libraries You can observe the hits.   Preferred solid supports include most of the synthetic, preferably sheet, spherical forms Polymer resins are included, that is, for example, polystyrene, polyolefins, Polymethyl methacrylates and similar resins, their derivatives and These are resins such as copolymers. Polymers of various degrees of crosslinking may also be used. A preferred solid support is Merrifield resin, which contains 1% Is a copolymer of divinylbenzene and polystyrene. Generally preferred polymers The support is insoluble in most organic solvents, but can swell in certain solvents. further Other support rests may include glass, ceramic, or metallic materials, Moreover, you may have those surfaces.   Not surprisingly, the residues of the selected molecule bind or attach to the surface of the solid support. That the solid support has functional groups that can participate in the condensation reaction is important. Such functional groups generally include halides, unsaturated substituents, Acids, hydroxyls, amines, esters, thiols, siloxy, Aza, oxo, etc. will be involved.   Coupling and subsequent release of the solid phase synthesized lipoglycopeptide. For ease, a linker substituent may be used. Such linkers Well known in the art, they include polyamino, polycarboxy, polyester, Including rihalo, polyhydroxy, polyunsaturated substituents, or combinations thereof However, the present invention is not limited to these.   This linker was used to prepare and manipulate compounds attached to the library Set that does not adversely affect the reagents used in Is preferably unstable under the given conditions. More preferably this linker -Is labile to acid or photolabile. Preferred linkers include A halotrityl moiety that binds at least one of the substituents P to the support (e.g., chloro Trityl moiety), or alpha-halo (e.g., bromine) -alpha-methylphenacyl Part included. Not surprisingly, the linker is Various of the substituents present may be covalently linked to each other or may be It can be used for covalently bonding to those contained in the substituent.   Usually, the covalent bond is one or more amines, ethers, thioethers, Ester, thioester, amide, acetamido, phosphate, phosphonate , A phosphinate, or a functional group containing a sulfate linkage may be utilized. This Because some libraries contain a sugar moiety, one or more glycosidic linkages It is expected that a match exists, but that is not required. Specific libraries of the invention Wherein at least one of the substituents of each of the formulas is adjacent Is covalently bonded to at least one of the substituents. Preferably the substituent S Is the same sugar, at least one substituent of substituent P and at least one of substituent L Covalently attached to a substituent. Furthermore, one of the sugars of the substituent S is replaced with one of the lipids of the substituent L. Are covalently bonded, glycoside ether bond, phosphate, pyrophospho , Phosphinyl, phosphanyl, phosphonate, phosphonyl, phosphono, Phosphino, phosphanoacetate, phosphonyl formate, phosphoramidyl Phosphorothionate, phosphonyl sulfonate, or phosphonyl sulfonate It may include a bond or the like. Some examples of glycosyl linkages are shown in Figure 5b I have. Further, the substituent L is formed through an acetamido bond of the alpha-anomeric hydroxyl group. It is also preferred to have at least one sugar of the substituent S to be covalently linked to the lipid. Would be better. More specifically, at least one sugar of the substituent S is a C-3 Via the alpha-acetamide or alpha-propionamide portion of the acid group And is covalently bonded to one of the substituents P.   The combination of amino acid residues is used in this library and its preparation method. Can be However, in some situations, the amino acid of the substituent P, pepti Or polypeptides may be all hydrophilic amino acid residues, or Alternatively, hydrophilic amino acid residues may dominate. Also included in the substituent P Are all hydrophobic amino acid residues or hydrophobic amino acid residues In some cases, the group may be dominant. Amino acids, peptides and polypeptides Various combinations of peptides include both stereoisomers, analogs, or their Homologs (homologs) were included and are shown in FIG.   Substituent S may be one or more as shown as an exemplary sugar substituent in FIG. The above monosaccharide, disaccharide, or polysaccharide may be contained. When specified, Monosaccharides include hexoses, pentoses, their deoxy analogs, their dideos Xy analogs, their azide-substituted analogs, or their amino-substituted analogs Good. The disaccharide or polysaccharide of the present invention includes hexose, pen Tooses, their deoxy analogs, their dideoxy analogs, their Including zid-substituted analogs, their amino-substituted analogs, or combinations thereof Can be taken.   The present invention allows various potential covalent bonds between sugars. I Thus, alpha or base, such as cis-1,2 or trans-1,2 sequences It may have a stereochemical configuration of a ta shape. (1,6), (1,3), (1,4) ) And the like, which includes possible stereochemical configurations.   An entire host of distinct lipids can be used in the present invention. Such a fat The quality can be simple lipids or complexed in nature. It is 2-60 Carbon atoms, preferably 5-55, more preferably 10-40, most preferably 1 May be composed entirely of aliphatic substituents containing 5-30 carbon atoms, or May be partially configured. In the case of cholesterol-bound structures, Fatty acid chains would be useful. In addition, this lipid is branched or unbranched Alkyl or alkenyl group (optionally one or more lower alkoxy Group, a lower acyloxy group, a halogen, or an aryl group). Or a branched or unbranched acyl group (optionally one or more Substituted with a lower alkoxy group, a lower acyloxy group, a halogen, or an aryl group Or a lower alkoxycarbonyl group and optionally one Represents a lower alkenyl group, a lower alkoxy group, a lower acyloxy group, Lower alkoxyca substituted with an aryl, aryl, or 9-fluorenyl group A rubonyl group, or one or more lower alkyl groups, It can be silicon substituted with a alkoxy group or an aryl group. Further It is also possible to use simple or complex aromatic or heteroaromatic structures it can. Examples of aromatic structures include substituted or unsubstituted hydroxy Cyphenylacetic acid, hydroxyphenylpropionic acid, methyl benzylate, hydride Roxynaphthoic acid, hydroxy-diphenylcarboxylic acid, hydroxy-fluorene Carboxylic acids, aromatic carboxylic acids such as hydroxybenzamide; Substituted or unsubstituted hydroxybenzofuran, hydroxy Like quinoline, thiophene carboxylic acid, hydroxythianaphthene carboxylic acid Hetero Aromatic structures and similar structures include, but are not limited to. Go Examples of some aromatic lipids are shown in FIG. 5a.   An important aspect of the present invention is the preparation of solid phase lipoglycopeptide libraries. How to This method uses a chemical formula as defined above, ie, R-P-FG₁Less than One or more of the substituents R, P and FG₁The step of providing This material , Which is also a chemical formula as defined above, i.e., FG_TwoReacts with the second unit with -S-L By doing, the first functional group FG₁One of the substituents P having_TwoEquipped Can proceed the bond formation reaction that forms a bond with the sugar of the substituted substituent S Become.   Specific aspects of the present invention as described in greater detail in the Examples section of this specification In an embodiment, the resin is first immersed in an organic solvent that swells the resin before performing the bonding step. Is done. Functional group FG of resin-binding component₁Is protected, e.g. fluoren Protecting groups such as the methyloxycarbonyl group (Fmoc) must first be removed. No. Thus, this method will eliminate any protection present on the substituents P, S, or L. A step of removing the group would be required.   The coupling step for reacting each functional group may be performed by an activating reagent or a well-known reagent. Is preferably promoted by adding a coupling reagent. like this The reagent assists, for example, in forming a bond between the carboxylic acid group and the amine group.   After the desired condensation reaction has progressed, the solid support of the obtained lipoglycopeptide The carrier-bound library may be released from the solid support, or Screening may be performed with the body attached. Thus, one or that Using more than one probe, receptor, affinity binder, enzyme or whole cell Lipoglycopeptide library obtained for or more active substances Screening is particularly designed.   When this compound is released from the solid support, the substituent P is cleaved from the substituent R. To obtain the structural unit P-S-L, which is then recovered You.   In one of the methods described above, one or more of the substituents S, L and FG_TwoIs One or more substituents R, P and FG₁And a chamber in the solvent in which the activating reagent is present. Combine at warm. Functional group FG_TwoAre pentafluorophenyl esters and especially EEDQ , HAT Reacts with amino groups in the presence of activating reagents such as U, HBTU, PyBOP, or similar substances. If available, it may be a simple free carboxylic acid. Various functional groups Need only be present anywhere in this molecular unit, But is present especially in one of the substituents P.   Some naturally occurring entities represented by the general formula P-S-L are known, The present invention also contemplates novel conjugates comprising other elements of the general formula. And already Despite being a known compound, it is defined by the solid phase method or as defined here. Such existing groups formed by cleavage of the bond between such substituents R and P Known compounds are considered to be part of the present invention. Therefore, given by its general formula A special synthesis view of the compound is planned. Combination approach for synthesis Chi is the preferred route.   It should be noted that certain compounds of this general formula are present in the substituent P The number of amino acid residues will differ from their natural counterparts. Preferred compounds Means two or more amino acids in the sequence, more preferably four or more , Most preferably 6 or more amino acids. Substituent P is further May further comprise one or more peptides or polypeptides. They may be linked sequences or may be non-pe They may be separated by peptide parts.   Yet another polypeptide component has three or more amino acids in its sequence. And has two or more sugars, preferably three or more sugars Is also good.   As mentioned elsewhere, the lipid portion of the compounds in this library is very large. Various structures can be adopted. However, usually, the lipid substituents have 4-60 Containing carbon atoms, preferably 5-50, and even more preferably 10-30 carbon atoms Will be.   Further, the various structural units that can be prepared by the method of the present invention are: Even though it is consistent with the structure of the naturally occurring lipid-bound glycopeptide intermediate Yes, they do not have to match. In some cases, including simple lipids, the substituent L It would be an advantage not to include the undecaprenyl group.   Thus, in the present invention, a compound represented by the following general formula (I) is planned. In this equation,   L¹Contains a lipid substituent,   P¹Comprises one or more amino acids, peptides or polypeptides,   G¹, G^Two, G^Three, G^FourOr G^FiveAre each independently a substituted or unsubstituted, branched Or unbranched alkyl, alkoxy, alkenyl, C1-C8 acyl Group, acetyl group, alkoxycarbonyl group, hydroxyalkyl group, carboxy An alkyl group or a substituted or unsubstituted aromatic or heteroaromatic group A substituent, or hydrogen,   X¹, X^Two, X^Five, X⁶Or X⁷Are each independently oxyalkyl, amine, Ter, thioether, ester, thioester, amide, acyl, acetamide , Phosphate, phosphinate, pyrophosphate, sulfate, azide, It may be a functional group containing a hydroxyl group or hydrogen, provided that the functional group is azide, In the case of a hydroxyl group or hydrogen, there is no attached G substituent,   X^ThreeOr X^FourAre each independently an amine, ether, thioether, ester , Thioester, amide, acetamide, phosphate, phosphinate, pyro Phosphate may be a functional group containing a sulfate substituent,   y is 0, 1, 2, or 3,   m, n, v, w, or z may independently be 0, 1, 2, or 3;   q or u may be independently 1, 2, or 3, provided that the sum of q and u is 5 Not bigger,   However, this compound is 2-N-acetyl-1-α-O-allyl-4,6-O-isopropylidene Muramyl-L-alanyl-D-glutamine benzyl ester, (2R) -benzyl 2- [N- (2'- N-acetyl-1'-α-O-allyl-4 ', 6'-O-acetylmuramyl-L-alanyl) amino] -4- Cyanobutanoate, (2R) benzyl 2-N-acetyl-1'-α-O-allyl-4,6-O-isoprop Lopylidenemuramil-L-alanyl) amino] -4-cyanobutanoate, (2R, 2 "R) -B Benzyl 2- [N- [2'-N-acetyl-1'-α-O-[(N, N-diisopropylamino) (butoxy ) Phosphinyl] -4 ', 6'-di-O-acetylmuramyl-L-alanyl] amino] -4-cyano Butanoate, (2R, 2 "R) -benzyl 2- [N- [2'-N-acetyl-1'-α-O-[[2" -carbobe Nzoxy-2 "-pentyloxy) ethoxy] (benzyloxy) phosphoryl] -4 ', 6'-di -O-acetylmuramyl-L-alanyl] amino] -4-cyanobutanoate, (2R, 2 "R) -2- [N- [2'-N-acetyl-1'-α-O-[[2 "-carboxy-2"-(pentyloxy) ethoxy] h Droxyphosphoryl] muramyl-L-alanyl] amino] -4-cyanobutanoate Or transglycosylase activity of penicillin-binding protein or murG gene product N-acetylglucosaminyltransferase activity on any of the natural Not a substrate.   Specifically, the compound represented by the chemical formula (I) has a substituent X¹G¹, X^FiveG^Three, X⁶G^Five, Or X⁷G^Four Is a hydroxyl group, HOCH_ThreeCH_Two-, Acetamide, phthalimide, benzoyl, alkoxy Cibenzoyl, alkoxycarbonylalkyl, carboxyalkyl, pivalyl Group, or any substituent commonly found on hydrocarbons. Contains lipoglycopeptide. X¹G¹Or X^TwoG^TwoThe substituent is located at the 2- or 5-position of the ring Preferably, X^ThreeP¹Is preferably located in position 3 and X^FourL¹Is the first position of the ring Is preferable. The compound of formula (I) may further comprise one or more A peptide or polypeptide and one or more lipids. Preferably each peptide or polypeptide has three or It contains more amino acids and each lipid has 2-60 carbon atoms. Containing, saturated, unsaturated, or polyunsaturated, linear, branched, or cyclic aliphatic Aromatics containing one or more substituents, or substituted or unsubstituted It contains one or more aromatic or heteroaromatic substituents. What are lipids and sugars? , Glycoside ether linkage, phosphate, pyrophosphate, phosphinyl, Phosphanyl, phosphone Phosphate, phosphonyl, phosphono, phosphino, phosphanoacetate, phosphonyl Formate, phosphoramidyl phosphorothionate, phosphonyl sulfonate, Or X containing a phosphonylsulfonate linkage^FourLinked via a covalent bond of Good to be.   The invention is further described by the following specific examples. Already in the above These examples are not required to practice the invention as properly described. As such, these examples should not be construed as limiting the invention to any particular point. 6 Example-Preparation of sugar, lipid and glycolipid building blocks 6.1 Allyl 2-acetamide-2-deoxy-D-glucopyranoside (2)   In anhydrous allyl alcohol (400 ml) containing boron trifluoride etherate (2 ml), Cetamide-2-deoxy-D-glucose suspension (1) (22.1 g, 0.1 mole) was not stirred. Heat for 2 hours. The clear solution is cooled and left overnight at room temperature to remove the crystals. Evaporate to give the title compound as a lump (α: β ratio 4: 1). TLC ( Ethyl acetate, isopropyl alcohol, water 9: 4: 2) Rf 0.5. 6.2 Allyl 2-acetamide-4,6-O-benzylidine-2-deoxy-α-D-glucopyranoside (3 )   Compound (2) (5 g, 0.019 mol) is dissolved in DMF (30 ml). Benzaldehyde dimethy Ruacetate (5.7 ml, 0.038 mole) and p-toluenesulfonic acid (380 mg, 0.002 mole) Was added. The reaction is left at 70 ° C. overnight for 18 hours. The reaction mixture is Blended by the addition of water and heavy carbon to neutralize the resulting mixture Sodium acid is added and the product precipitates. The precipitate is filtered and washed with water. Washed well. Recrystallization from methanol / water gives pure α-isomer (3.2 g, 47%) . TLC (10% methanol / methylene chloride) Rf 0.7. 6.3. Allyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri-O-acetyl-2'-deoxy C-β-D-glucopyranosyl) -2-deoxy-3-O [1- (R)-(methoxycarbonyl) ethyl Ru] -α-D-glucopyranoside (8)   Compound (6) (150 mg, 0.43 mole) and compound (7) (214 mg, 0.65 mmol) And dried twice by azeotropic distillation from, and finally dissolved in toluene (10 ml). This To this mixture is added p-toluenesulfonic acid (10 mg). Heated at 110 ° C for 3 hours . The solvent is concentrated under reduced pressure and the mixture is passed through a silica gel column. Methylenek Eluted with methylene chloride, and then the ratio of methanol in methylene chloride was Elution is performed by increasing the concentration to 5% methanol in chloride to obtain a product (200 mg , 70%). TLC (10% methanol / methylene chloride) Rf 0.6. 6.4. Allyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri-O-acetyl-2'-deoxy C-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3-O [1 (R)-(methoxycal Bonyl) ethyl] -α-D-glucopyranoside (9)   Compound (8) (200 mg, 0.3 mmol) is dissolved in pyridine (2 ml). For this solution Acetic anhydride (1 ml) and DMAP (10 mg) are added. The reaction is stirred at room temperature for 6 hours. And evaporated under reduced pressure. The residue was recovered with water (10 ml) and extracted twice with ethyl acetate. It is. The organic layer extract is washed with water and saturated sodium chloride solution, Dry with a filter and filter. After evaporation of the solvent, the crystalline product (210 mg , 100%). TLC (10% methanol / methylene chloride) Rf 0.62. 6.5. Carboxy-2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri-O-acetyl-2'-de Oxy-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3-O [1 (R)-(methoxy Carbonyl) ethyl] -α-D-glucopyranoside (10)   Compound (9) (200 mg, 0.28 mmol) is dissolved in the solvent mixture: carbon tetrachloride (0.8 ml), Acetonitrile (0.8 ml), and water (1.2 ml). This mixture is sodium periodate (250 mg, 1.17 mol) and ruthenium (III) chloride (2 mg). This reaction mixture The mixture is stirred at room temperature for 2 hours. NaIO_ThreeIs observed. The mixture is CH_TwoCl_Two(2 x 15 ml). The organic layer is washed once with water and Na_TwoSO_FourAnd dry. After filtration The solvent is evaporated to give the product (180 mg, 86%) as a solid. Rf0.1 . 6.6. Dodecylcarbamoylmethyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri- O-acetyl-2'-deoxy-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3- O- [1 (R)-(methoxycarbonyl) ethyl] -α-D-glucopyranoside (11)   Compound (10) (700 mg, 0.95 mmol), 1-hydroxybenzotriazole (142 mg, 1.0 5 mmol), 1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide meso-p- Toluenesulfonic acid (445 mg, 1.05 mmol) (WSCDI), and dodecylamine (210 mg, 1.14 mmol) is stirred in methylene chloride (40 ml) for 30 hours. The reaction mixture is Dilute with methylene chloride (60 ml), wash with water, saturated NaCl solution, then add sodium sulfate Dry with lium. The organic layer is filtered and the solvent is removed under vacuum. This residue To obtain the product (620 mg, 73%) as a crystalline solid, CH_TwoCl_Two-7% EtOH / CH_TwoCl_TwoSolvent Purified by chromatography on silica using an elution gradient. TLC (10 % Methanol / methylene chloride) Rf 0.6. 6.7. Dodecylcarbamoylmethyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri- O-acetyl-2'-deoxy-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3- O- [1 (R)-(carboxy) ethyl] -α-D-glucopyranoside (12)   Compound (11) (100 mg, 0.11 mmol) was dissolved in 1,4-dioxane (10 ml), and the Stirred. A 0.1 M aqueous potassium hydroxide solution (1.7 ml, 0.17 mmol) was added to the reaction solution. And stirred at 0 ° C. for 4 hours. The reaction mixture is neutralized with Amberlite H + resin Is completed. The reaction mixture was filtered and the solvent was the product as a solid (90 mg, 91% ). TLC (20% methanol / methylene chloride) Rf 0.45.6.8. Dodecylcarbamoylmethyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri- O-acetyl-2'-deoxy-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3- O- [1 (R)-(pentafluorophenoxycarbonyl) ethyl] -α-D-glucopyranosi Do (13)   Compound (12) (50 mg, 0.056 mmol) is dissolved in methylene chloride (10 ml). this Pentafluorophenol (12 mg, 0.067 mmol) and 1-cyclohexyl-3- were added to the solution. (2-morpholinoethyl) carbodiimide meso-p-toluenesulfonate (29 mg , 0.067 mmol) are added. The mixture is stirred at room temperature for 2 days. Reaction mixture Diluted with methylene chloride and washed well with ice-cold water and ice-cold saturated NaCl solution. And then dried over sodium sulfate. The mixture is filtered and the solvent is solid. And evaporated to dryness to give the product (53 mg, 89%). TLC (ethyl acetate: Seton 4: 1) Rf 0.7. WSCD1 = 1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide         Meso-p-toluenesulfonic acid 6.9. Allyl 2-acetamide-4,6-O-benzylidene-2-deoxy-3-O- [methoxycarbonyl Methyl] -α-D-glucopyranoside (14)   Compound (3) (349 mg, 1 mmol) was dissolved in THF (10 ml) and to this solution was added 60% NaH in oil. (80 mg, 2 mmol) is added. The mixture is stirred at room temperature under argon for 30 minutes. To this mixture is added methyl bromoacetate (0.19 ml, 2 mmol). The mixture is stirred at 50 ° C. under argon for 23 hours. Acetic acid (2 mmol) stopped the reaction. Added to stop. The solvent is evaporated and water (10 ml) is added to the residue . After stirring, the precipitated product is filtered and dried (420 mg, 100%). TLC (10% methano Methylene chloride) Rf 0.55. 6.10a. Allyl 2-acetamide-2-deoxy-3-O- [methoxycarbonylmethyl] -α-D-gluco Pyranoside (15)   Compound (14) (1.5 g, 3.6 mmol) was treated with 75% acetic acid / water and the product ( (810 mg, 67%) is processed as described for compound (6). TLC (10% methanol / methylene chloride) Rf 0.4. 6.10b. Allyl 2-acetamide-4,6-di-O-acetyl-2-deoxy-3-O- [methoxycarbonyl Methyl] -α-D-glucopyranoside (14)   As in Example (9), to obtain the product (21) (500 mg, 53%) as a solid, the compound ( 15) (750 mg, 2.25 mmol) treated with acetic anhydride and pyridine overnight with catalytic amount of DMAP Is done. TLC (ethyl acetate: acetone 4: 1) Rf 0.65. 6.11. Allyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri-O-acetyl-2'-deoxy (Β-D-glucopyranosyl) -2-deoxy-3-O- [methoxycarbonylmethyl] -α-D -Glucopyranoside (16)   To obtain compound (16) (3.0 g, 68%), the compound was prepared in the same manner as for compound (8). Compound (15) (2.2 g, 6.6 mmol) and compound (7) (3.5 g, 10.6 mmol) were dissolved in toluene (150 ml). Heated with toluenesulfonic acid (150 mg). TLC (10% methanol / methylene Chloride) Rf 0.6. 6.12. Allyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri-O-acetyl-2'-deoxy (3-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3-O- [methoxycarbonyl Rumethyl] -α-D-glucopyranoside (17)   Compound (16) (3.0 g, 4.5 mmol) was treated with pyridine (30 m l), acetic anhydride (15 ml), and DMAP (30 mg). Recrystallized with ethyl acetate After work-up, pure compound (17) (1.1 g, 36%) is obtained. TLC (ethyl acetate) Rf0.2 . 6.13. Carboxymethyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri-O-acetyl -2'-deoxy-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3-O- [methoxy [Cicarbonylmethyl] -α-D-glucopyranoside (18)   To obtain compound (18) (333 mg, 77%) as a solid, as in compound (10) Compound (17) (420 mg, 0.6 mmol) was treated with sodium periodate (510 mg, 2.4 mmol) and And treated with ruthenium (III) chloride. TLC (10% methanol / methylene chloride) Rf0.1.6.14. Dodecylcarbamoylmethyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri- O-acetyl-2'-deoxy-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3- O- [methoxycarbonylmethyl] -α-D-glucopyranoside (19)   As with compound (11), compound (18) (330 mg, 0.46 mmol) was Nzotriazole (74 mg, 0.55 mmol), 1-cyclohexyl-3- (2-morpholinoethyl Carbodiimide meso-p-toluenesulfonic acid (292 mg, 0.69 mmol), and Treated with decylamine (102 mg, 0.55 mmol). After silica gel column purification, Compound (19) (250 mg, 61%) is obtained as a solid. TLC (10% methanol / methylene chloride Lido) Rf0.6. 6.15. Dodecylcarbamoylmethyl 2-acetamide-6-O- (2'-acetamide-3 ', 4', 6'-tri- O-acetyl-2'-deoxy-β-D-glucopyranosyl) -4-O-acetyl-2-deoxy-3- O- [carboxymethyl] -α-D-glucopyranoside (19)   As in the case of compound (12), compound (19) (250 mg, 0.28 mmol) was treated with dioxane (12 ml )) (4.2 ml, 0.42 mmol). After completion of the reaction, compound (20) (23 (0 mg, 94%) is obtained as a solid. TLC (20% methanol / methylene chloride) Rf0.45 .6.16. Carboxymethyl 2-acetamide-4,6-di-O-acetyl-2-deoxy-3-O- [methoxy Carbonylmethyl] -α-D-glucopyranoside (22)   To obtain compound (22) (540 mg, 70%) as a solid, use the same method as for compound (10). By the method, compound (21) (740 mg, 177 mmol) was treated with sodium periodate (1.5 g, 7.08 mol) And with ruthenium (III) chloride. TLC (10% methanol / methylene chloride Lido) Rf0.1. 6.17. Dodecylcarbamoylmethyl 2-acetamide-4,6-di-O-acetyl-2-deoxy-3-O- [Methoxycarbonylmethyl] -α-D-glucopyranoside (23)   As in the case of compound (11), compound (22) (540 mg, 1.2 mmol) Zotriazole (195 mg, 1.44 mmol), 1-cyclohexyl-3- (2-morpholinoethyl B) carbodiimide meso-p-toluenesulfonic acid (762 mg, 1.8 mmol), and Treated with decylamine (267 mg, 1.44 mmol). Silica gel column and ethyl acetate After elution with, the purified compound (23) (450 mg, 63%) is obtained as a solid. TLC (10% meta (Nor / methylene chloride) Rf 0.6. 6.18. Dodecylcarbamoylmethyl 2-acetamide-4,6-di-O-acetyl-2-deoxy-3-O- [Carboxymethyl] -α-D-glucopyranoside (23)   As in the case of compound (12), compound (23) (150 mg, 0.5 mmol) was treated with dioxane (10 ml). 0.1 M KOH aqueous solution (3.8 ml, 0.38 mmol), and after completion of the reaction, compound (20) (148 mg, 100%) is obtained as a solid. TLC (10% methanol / methylene chloride) Rf0.1 . 6.19. Dodecylcarbamoylmethyl 2-acetamide-4,6-di-O-acetyl-2-deoxy-3-O- [Pentafluorophenoxycarbomoylmethyl] -α-D-glucopyranoside (25) As with compound (13), compound (24) (300 mg, 0.5 mmol) Phenol (101 mg, 0.55 mmol) and 1-cyclohexyl-3- (2-morpholinoethyl) Treated with rubodiimide meso-p-toluenesulfonic acid (282 mg, 0.65 mmol). Anti After completion of the reaction, compound (25) (450 mg, 100%) is obtained as a solid. TLC (10% methanol / Methylene chloride) Rf 0.7. 6.20. Allyl 2-acetamide-4,6-O-benzylidene-2-deoxy-3-O- [carbonylmethyl] -α-D-glucopyranoside (47)   Compound (14) (1.0 g, 2.4 mmol) is dissolved in methanol (40 ml). Add water to this solution Potassium oxide (403 mg, 7.2 mmol) and water (0.1 ml) were added and stirred at room temperature for 4 hours. It is. This alkaline mixture is Amberlite H⁺And filtered. Dissolution After evaporation of the medium, a crystalline product is obtained (100%). TLC (10% methanol / methyl Len chloride) Rf 0.1. 6.21. Allyl 2-acetamide-4,6-O-benzylidene-2-deoxy-3-O- [pentafluorofe [Noxycarbomoylmethyl] -α-D-glucopyranoside (48)   As with compound (13), compound (47) (1.0 g, 2.4 mmol) Enol (497 mg, 2.7 mmol) and 1-cyclohexyl-3- (2-morpholinoethyl) Treated with carbodiimide meso-p-toluenesulfonic acid (1.14 g, 2.7 mmol). Anti After completion of the reaction, compound (48) (1.4 g, 100%) is obtained as a solid. TLC (10% methanol / Methylene chloride) Rf 0.75. 6.22 Methyl 2 (S) -O-trifluoromethanesulfonyloxypropiolate (4)   To a stirred solution of anhydrous pyridine (6.7 ml, 83 mmol) in anhydrous dichloromethane (250 ml) Trifluoromethanesulfonyl anhydride (13.3 ml, 79 mmol) was added dropwise at -40 ° C. To give a white suspension. Thereafter, methyl-2 (S) -hexane dissolved in dichloromethane (15 ml) was used. Droxypropiolate (8.2 g, 79 mmol) is added to the reaction mixture at -40 ° C and stirred The reaction mixture is allowed to warm to room temperature over 1 hour. White The color precipitate is removed by filtration and the filtrate is rotary evaporated until a brown solution is obtained. It is concentrated at 30 ° C or less using a rotator. This liquid passes through a 15 cm plug of silica. (P_TwoO_Five(Eluted with 250 ml of hexane dried) to give a pale yellow liquid (9.8 g). And concentrated until a crude product is obtained. Pure methyl (S) -2-O-tri Fluoromethanesulfonyl loopropiolate (4) (7.8 g, 33 mmol, 42% yield) Obtained as a colorless, free-flowing liquid (boiling point 60 ° C., 10 torr). 6.23. Allyl 4,6-O-benzylidene-3-O- [1 '(R)-(methoxycarbonyl) ethyl] -2-deoxy Ci-2-acetamide-α-D-glucopyranoside (5a) and allyl-4,6-O-benzylidene -3-O- [1 '(S)-(methoxycarbonyl) ethyl] -2-deoxy-2-acetamide-α-D-g Lucopyranoside (5b)   Stirred suspension of alcohol (3) (9.06 g, 26 mmol) in anhydrous dichloromethane (150 ml) Was heated at 40 ° C. for 10 minutes under argon, then 95% sodium hydride (0.78 g, 32.5 m mol). After 5 minutes, triflate (4) (9.2 g, 39 mmol) was added with vigorous exotherm and hydrogen. Is carefully added over 15 minutes, causing purification of the compound. Another 20 minutes Later, the reaction mixture is cooled and poured into water (100 ml). Separate the organic layer, Wash with water (50ml), Na_TwoSO_FourDried and evaporated to give a white solid (15g). The crude product was purified by flash chromatography on silica (250 g) Purified using a gradient elution of 0 → 20% ethyl acetate-dichloromethane, first allyl 4,6-O-benzylidene-3-O- [1 ′ (R)-(methoxycarbo) xyethyl] -2-deoxy 2-acetamide-α-D-glucopyranoside (5a) (6.1 g, 14 mol, 54% yield), followed by Lyl-4,6-O-benzylidene-3-O- [1 ′ (S)-(methoxycarboxy) ethyl] -2-deoxy Ci-2-acetamide-α-D-glucopyranoside (5b) (1.65 g, 3.8 mmol, 14.5% yield) Both were obtained as white solids.   Spectral data for 5a: TLC (30% EtOAc-hexane) Rf 0.55. IR (KBr disk C):  Spectral data for 5b: TLC (30% EtOAc-hexane) Rf 0.38. IR (KBr disk C): 6.24. Allyl 3-O- [1 ′ (R)-(methoxycarbonyl) ethyl] -2-deoxy-2-acetamide-α -D-glucopyranoside (6)   75% AcOH-H_TwoIn O (100 ml), benzylidene acetal (5a) (8.0 g, 8.4 mmol) was stirred and dissolved. The liquid is heated at 55 ° C. for 14 hours. The reaction mixture is concentrated under reduced pressure at 50 ° C. Dry (2 x 100 ml) by azeotropic distillation. Crude product is pre-adsorbed on silica (25 g) Flash chromatography on silica (eluent is 10 → 20% ethanol Chloromethane) and purified as an oil in the form of allyl 3-O- [1 '(R)-(methoxy). [Cicarbonyl) ethyl] -2-deoxy-2-acetamide-α-D-glucopyranoside (6) (4. 65 g, 13.4 mmol, 83% yield) are obtained. TLC (10% MeOH-CH_TwoCl_Two) RF 0 .52. IR (pure): 6.25. 2-methyl-4,5- (tri-O-acetyl-2-deoxy-α-D-glucopyranosyl) -Δ^Two-Oki Sazolin (7)   2-Deoxy-2-acetamide-α-D-glucopyranoside (49.0 g, 222 mmol) Stir in cetyl (90 ml, 1.261 mol) for 18 hours at room temperature. This clear red oil Diluted with ethanol-free chloroform (400 ml) and then poured onto ice (700 ml) You. The organic layer was separated and ice cold saturated NaHCO_ThreeWashed quickly with aqueous solution (500 ml), N o_TwoSO_FourAnd concentrated under reduced pressure to a volume of about 50 ml. Diethyl ether (400m l) is added and the product 3 ', 4', 6'-tri-O-acetyl-2'-deoxy-2'-acetami Do-α-D-glucopyranosyl chloride (58 g, yield 71%) is an off-white crystal. And collected. The chloride (40 g, 109 mmol) and anhydrous sodium bicarbonate (18.3 g , 218 mmol) in anhydrous acetonitrile (80 ml)._TwoUntil gas generation stops (about 30 minutes ) Stir. The reaction mixture is filtered and the filtrate is evaporated. Residue is dicro Dissolved in dichloromethane (300ml), washed with water (4x50ml) and evaporated again. Remaining The residue is collected with diethyl ether (250 ml) and stored at -20 ° C for 3 hours. Colorless Pure crystal, 1,3,4,6-tetra-O-acetyl-2-deoxy-2-acetamide-D-g Lucopyranoside (2.5 g) is removed by filtration. The product, 2-methyl-4,5- (tri -O-acetyl-2-deoxy-α-D-glucopyranosyl) -Δ^Two-Oxazoline (7) (29 g, 88 mmol, 81% yield) are obtained by evaporation of the filtrate as a golden yellow oil. TLC (Et20) Rf 0.24. 6.26. Carboxymethyl 4,6-O-benzylidene-3-O- [1 '(R)-(methoxycarbonyl) ethyl ] -2-Deoxy-2-acetamide-α-D-glucopyranoside (26)   Carboxylic acid (26) was compound (5a) (1.28 g, 2.94 mm) in the same manner as for compound (10). ol) and sodium periodate (2.83 g, 13.24 mmol) and ruthenium (III) trichloride Prepared by reaction of hydrate (12 mg). The crude oil (1.8 g) is Used without further purification.6.27. Dodecylcarbamoylmethyl 4,6-O-benzylidene-3-O- [1 '(R)-(methoxycarbo Nyl) ethyl] -2-deoxy-2-acetamide-α-D-glucopyranoside (27)   A solution of the unpurified acid (26) (1.8 g, 2.9 mmol), 1-cyclohexyl-3- (2-mol Folinoethyl) carbodiimide meso-p-toluenesulfonic acid (WSCDI) (1.49 g, 3 .53 mmol), HOBt (0.476 g, 3.53 mmol) and dodecylamine (0.598 g, 3.23 mmol) Stir in dichloromethane (100 ml) at room temperature for 15 hours. The solvent is evaporated and this The residue was collected with EtOAc (100 ml), 2N HCl (50 ml), saturated NaHCO_ThreeWash with aqueous solution (50 ml) And then Na_TwoSO_FourAnd dry. Evaporation under reduced pressure gave a white solid (4.0 g). This was flash chromatographed on silica eluting with 3% MeOH-DCM. Afforded the title compound (27) as a white solid (0.97 g, 54% yield over two steps). You. 6.28. Dodecylcarbamoylmethyl 4,6-O-benzylidene-3-O- [1 '(R)-(carboxy) ethyl Ru] -2-deoxy-2-acetamide-α-D-glucopyranoside (28)   A solution of the methyl ester (27) (0.97 g, 1.56 mmol), and sodium hydroxide (187 m g, 4.69 mmol) is stirred at room temperature in 1: 1 THF-MeOH (50 ml) for 14 hours. The solution is Box (Dowex) 50H⁺Acidified with resin, filtered and evaporated to give a pale yellow solid You.6.29. Dodecylcarbamoylmethyl 4,6-O-benzylidene-3-O- [1 '(S)-(methoxycarbo Nyl) ethyl] -2-deoxy-2-acetamide-α-D-glucopyranoside (29)   The allyl group (5b) (2.00 g, 4.60 mmol) of the compound (5b) is sodium periodate (4.43 g, 20.70 mmol) and ruthenium trichloride hydrate (17 mg) to give compound (10). As in the case of the preparation, it is substituted with a methylene carboxy group, and To obtain a crude product acid. TLC (10% MeOH-DCM) Rf is the position of the reference line. Crude The resulting acid is then 1-cyclohexyl-3, as in the preparation of compound (27). -(2-morpholinoethyl) carbodiimide meso-p-toluenesulfonic acid (2.34 g, 5.52 mmol), HOBt (0.745 g, 5.52 mmol) and dodecylamine (0.936 g, 5.06 mmol) To be able to react with. After reaction and chromatography, the amide (29) (0.8 g, 1.29 mmol, 28% yield) as a white solid. 6.30 Dodecylcarbamoylmethyl 4,6-O-benzylidene-3-O- [1 '(S)-(carboxy) ethyl Ru] -2-deoxy-2-acetamide-α-D-glucopyranoside (30)   A solution of methyl ester (29) (0.80 g, 1.29 mmol) and sodium hydroxide (154 mg , 3.87 mmol) are stirred at room temperature in 1: 1 THF-MeOH (50 ml) for 14 hours. This solution is Box (Dowex) 50H⁺Acidified with resin, filtered and evaporated to a white solid To give the carboxylic acid (30).6.31. Allyl 4,6-O-benzylidene-3-O-benzyl-2-deoxy-2-acetamide-α-D-glu Copyranoside (31)   Alcohol (3) (40.5 g, 106.7 mmol) is azeotroped with toluene (100 ml) and then alcohol Dissolve in anhydrous DMF (700 ml) under a gon. 95% NaH (3.23 g, 28.lmmol) is added in portions over 5 minutes. After the production of hydrogen gas stops, Benzyl bromide (15.3 ml, 128.l mmol) was added in one portion and the reaction mixture was flushed with argon. Stir at room temperature overnight. The reaction mixture was poured into water (1 L) and the resulting crude product was white. Precipitates as a colored solid. This is collected by filtration and the mass on the filter paper is water (3x500 ml) and then washed with hexane (2x200ml). Remove the lump on the wet filter paper with DCM ( 1 L), washed with brine (100 ml),_TwoSO_FourBy drying and evaporating Benzyl ether (31) (42.33 g, 90.3% yield) as an off-white solid. Can be 6.32. Allyl 3,6-di-O-benzyl-2-deoxy-2-acetamide-α-D-glucopyranoside (32 )   Benzylidene acetal (31) (42.33 g, 96.4 mmol) was once co-existed with toluene (150 ml). Boil and then dry under vacuum in a 2 L flask. Molecular sieve made into 3A powder (Molecular sieves) (65 g), sodium cyanoborohydride (54.5 g, 868 mmol) And anhydrous THF (800 ml) is added under argon. The stirred suspension is cooled to 0 ° C 1N HCl in diethyl ether (800 ml) was added via cannula over 40 minutes Is done. The milky solution was TLC (10% MeOH-CH_TwoCl_Two) Terminates the reaction. Stir at room temperature until done. 2N HCl (aqueous solution) (300 ml) was added and the reaction mixture was The liquid is shaken and then ethyl acetate (1 L) is added. The organic layer is separated and reduced Concentrated under reduced pressure to a yellow liquid (200 ml), then redissolved in dichloromethane (1 L) It is. Ethanolamine (70 ml) is added and the reaction mixture is stirred for 20 minutes at room temperature. , Then washed with water (500ml) and with 2N HCl (aq) (2x500ml) and brine (100ml) Washed. Na_TwoSO_FourThe title compound (32) (41 g, yield 95%) as an orange oil and a few days to become an amorphous white solid Crystallized over. 6.33. 2-phthalimido-2-deoxy-1,3,4,6-tetra-O-acetyl-β-D-glucopyranoside ( 33)   Compound (33) (23 g, 48.2 mmol) was obtained from Remu (R.U.Lemieux), Takeda (T. Takeda), And B.Y. CHang, Am. Chem. Soc. Symp. 39, 90, and (1976) Is prepared. White crystalline solid. TLC (50% EtOAc-hexane) Rf 0.24. 6.34 Allyl 4-O- (2'-phthalimido-2'-deoxy-3 ', 4', 6'-tri-O-acetyl-β-D-glu (Co-44 pyranosyl) -3,6-di-O-benzyl-2-deoxy-2-acetamide-α-D-gluco Pyranoside (34)   Compound (32) (14.8 g, 33.6 mmol) and compound (33) (22.4 g, 46.9 mmol) were treated with anhydrous toluene. Powdered 4A sieve co-azeotroped in ene (120 ml), then activated And the mixture is resuspended in anhydrous dichloromethane (500 ml). The suspension is Stir for 1 h under argon, after which TMSOTf (35 ml) is added to the stirred suspension A second 10 ml is added after 30 minutes and a further 4 hours later at 10 ml. The reaction is It is stopped after 4 hours 30 minutes by the addition of triethylamine (70 ml). The black solution Filtered through celite pad, then washed with dichloromethane (300ml) . The combined filtrate was water (2x300ml), 2N HCl (aq) (2x200ml) and Wash once with brine (200 ml), then dry (Na_TwoSO_Four), Until a brown solid (33 g) To evaporate. Use a gradient elution of 50% to 100% EtOAc: Hexane to filter on silica. Rush chromatography yields reaction product (33) (4.2 g) as a yellow oil. The title compound (17.4 g, 60.5% yield) is obtained. 6.35. Allyl 4-O- (2'-phthalimido-2'-deoxy-β-D-glucopyranosyl) -3,6-di-O- Benzyl-2-deoxy-2-acetamide-α-D-glucopyranoside (35)   Compound (34) (17.4 g, 20.3 mmol) was dried by azeotropic distillation from toluene (50 ml). And then dissolved in anhydrous methanol (150 ml) in the presence of argon. Sodium meth Oxide (1.1 g) is added and the reaction mixture is stirred at room temperature for 1 hour (TLC: 10% MeOH-DC The reaction was terminated by M). Subsequently, 1N HCl in diethyl ether (15 ml) was mixed. The solution was added to adjust the pH of the solution to pH 5, and concentrated to give a brown foam (20 g). To give the title compound (35).6.36. Allyl 4-O- (3 ', 4', 6'-tri-O-acetyl-2'-acetamide-2'-deoxy-β-D-glu (Copyranosyl) -3,6-di-O-benzyl-2-deoxy-2-acetamide-α-D-glucopyra Noside (36)   Unpurified compound (35) (20 g, 20.3 mmol) was dissolved in ethylenediamine in ethanol ( Heat at 75 ° C. for 16 h under argon with 4 equivalents of 4.86 g (81.2 mmol). reaction The mixture was evaporated under reduced pressure at 65 ° C and then dried by azeotropic distillation from toluene (100 ml). Drying gives the free amine as a brown solid. This solid is anhydrous vinegar A solution containing acid (35 ml), pyridine (60 ml), and 4-dimethylaminopyridine (0.2 g) Resuspended in dichloromethane (60 ml) and the temperature of the reaction mixture rises to 45 ° C. Reaction mixture Compound Stir at room temperature for 24 hours, then dilute with dichloromethane (400 ml) and add ice cold water (2x40 Oml), ice-cold sodium bicarbonate (2x200ml), 2N HCl (2x200ml), then brine (100ml) Washed with. The solution is Na_TwoSO_FourAnd evaporated to a brown solid (15.93 g). Can be done. Use a 2% → 5% Me0H-DCM solvent elution gradient to flash flash on silica. The title compound (12.58 g, 78% yield) was obtained as a white solid by chromatography. It is.6.37. Dodecylcarbamoylmethyl 4-O- (3 ', 4', 6'-tri-O-acetyl-2'-acetamide-2 ' -Deoxy-β-D-glucopyranosyl) -3,6-di-O-benzyl-2-deoxy-2-acetami De-α-D-glucopyranoside (37)   3 Constituent solvents: vigorously in acetonitrile (1 ml), carbon tetrachloride (1 ml), and water Stirred compound (36) (0.49 g, 0.69 mmol) and ruthenium trichloride hydrate (3mg, 1x10^-FiveSodium periodate (0.61 g, 2.83 mmol) added to an ice-cold suspension of (M) Is done. After 10 minutes, the stirred reaction mixture is allowed to warm to room temperature for 2 hours Is done. The reaction mixture is diluted with dichloromethane (50ml) and filtered through celite And the filtrate is evaporated to a tan solid (0.43 g).¹H NMR (CDCl_Three) This solid is mainly a 1-carboxymethyl derivative due to incomplete oxidation It is suggested that a small amount of the 1-formylmethyl derivative is mixed in. FAB MS (relative to carboxylic acid): 811 (M + Na)⁺, 833 (M + 2Na-H)⁺. The mixture is further Used for next step without purification.   Unpurified carboxylic acid (0.43 g, about 0.54 mmol), dodecylamine (110 mg, 0.5 mg 9 mmol), 1-hydroxybenzotriazole (80 mg, 0.59 mmol), and dicyclohexane Xylcarbodiimide (DCC) (133 mg, 0.65 mmol) in dichloromethane (15 ml) at room temperature For 20 hours. The reaction mixture was filtered, diluted with dichloromethane (40 ml), 2N H Wash with Cl (2x20ml), sodium bicarbonate solution (2x20ml) and brine (20ml), then Dry (Na_TwoS0_Four) Evaporation gives a tan solid (0.52 g). CH_TwoCl_Two → 3% MeOH-CH_TwoCl_TwoFlash chromatography on silica (15 g) using a solvent elution gradient of By chromatography, the title compound (37) as a white solid (380 mg, 0.39 mmol, yield 58%). 6.38. Dodecylcarbamoylmethyl 4-O- (3 ', 4', 6'-tri-O-acetyl-2'-acetamide-2 ' -Deoxy-β-D-glucopyranosyl) -2-deoxy-2-acetamide-α-D-glucopyr Noside (38)   A solution of compound (37) (4.2 g, 4.4 mmol) in ethanol (50 ml) and 10% Pd / C (0.75 g) Is shaken at room temperature in a Parr hydrogenator under 50 PSI of hydrogen gas for 3 days. The product crystallized from the reaction mixture was re-dissolved by the addition of dichloromethane (300 ml). The catalyst is removed and the catalyst is removed by filtration through a pad of celite. White by evaporation Solid (^-5 g) are obtained. This solid is 5% → 15% MeOH-CH_TwoCl_TwoUse a solvent elution gradient of And purified by flash chromatography on silica to give a white solid. The title compound (38) (2.3 g, 67% yield) is obtained. 6.39. Dodecylcarbamoylmethyl 4-O- (3 ', 4', 6'-tri-O-acetyl-2'-acetamide-2 ' -Deoxy-β-D-glucopyranosyl) -6-O-benzoyl-2-deoxy-2-acetamide- α-D-glucopyranoside (39)   Benzoyl chloride (0.75 ml, 6.5 mmol) was obtained at -20 ° C with (4: 1) pyridine-dichlorometh Compound (38) (2.8 g, 3.61 mmol) and DMAP (0.2 g) are stirred in tan (100 ml) Added to the solution. The reaction mixture is left to warm to room temperature for 1 hour. You. The reaction mixture was cooled again to -20 ° C and DMF (20 mL) was added. It becomes transparent and bright yellow. The reaction is left to warm to room temperature for 1 hour. And then diluted with dichloromethane (300ml), water (300ml), 2N HCl (4x200ml), Wash with sodium bicarbonate (100 ml), and brine (100 ml). This solution is Na_TwoS0_Four And dry. Methanol (30 ml) was added and the solution was filtered and evaporated A white solid (4 g) is obtained. Flash chromatography on silica, solvent: 5% → 15% MeOH-CH_TwoCl_TwoAfforded the title compound (39) (1.6 g, 50.5% yield) as a white solid. And diol (38) (1.1 g, 39% recovery). Compound (39) 6.40. Dodecylcarbamoylmethyl 4-O- (3 ', 4', 6'-tri-O-acetyl-2'-acetamide-2 ' -Deoxy-β-D-glucopyranosyl) -6-O-benzoyl-3-O-methoxycarbonyl Cyl-2-deoxy-2-acetamide-α-D-glucopyranoside (40)   Compound (39) (300 mg, 0.34 mmol) and 18-crown-6 were added in anhydrous THF (30 ml). Add 0.5M potassium hexamethyldisocyanate in toluene A razide (1.7 ml, 0.85 mmol) solution is added. The reaction mixture is stirred for 10 minutes, Followed by methyl 2-O-trifluoromethanesulfonyloxyacetate (compound (4) (Prepared from methylglucolate in a similar manner) (300 mg, 1.36 mmole). Is added to this. The reaction mixture was stirred for 3 hours at 50 ° C and stopped with 2N HCl (0.5ml). Is stopped. Evaporation gives a yellow oily residue which is Fractionated between dichloromethane (75 ml) and water (70 ml), dried (Na_TwoSO_Four), Evaporate Can be Flash chromatography on silica using a 2% → 10% MeOH-DCM solvent elution gradient The first fraction (80 mg), TLC (5% MeOH-DCM) Rf 0.35 and And a fraction (120 mg) after co-action with compound (39), TLC (5% MeOH-DCM) Rf 0.25. Most 974 (M + Na) for the first fractionated FAB MS⁺And 1046 (dialkylated product + Na)^-With mass p Is obtained. No initial material is seen in this first fractionation. FAB MS in later fractionation Is 934 (M + Na-Ac)⁺And 902 (initial material + Na)⁺Gives a mass peak. 6.41. Dodecylcarbamoylmethyl 4-O- (3 ', 4', 6'-tri-O-acetyl-2'-acetamide-2 ' -Deoxy-β-D-glucopyranosyl) -6-O-benzoyl-3-O-carboxymethyl-2- Deoxy-2-acetamide-α-D-glucopyranoside (41)   A 0.1 M aqueous solution of potassium hydroxide (2.0 ml, 0.2 mmol) was added to the compound (3 9) (120 mg, 0.12 mmol) added to a stirred ice-cold solution, the reaction mixture consumes the initial material (10% MeOH-DCM by TLC, ca. 4 h). This solution is Bar Light H⁺Neutralized with resin and evaporated. The residue was prepared from toluene (20 ml). Drying by boiling distillation gives the unpurified title compound (41) (110 mg) . 6.42. Dodecylcarbamoylmethyl 4-O- (3 ', 4',-di-O-acetyl-2'-acetamide-2'-de Oxy-β-D-glucopyranosyl) -6-O-benzoyl-3-O- [1 (R)-(methoxycarbonyl Ru) ethyl] -2-deoxy-2-acetamide-α-D-glucopyranoside (42)   As in Example (40), compound (39) (100 mg, 0.114 mmol) was potassium hydroxide in toluene. 0.5M solution of umhexamethyldisilazide (0.46 ml, 0.23 mmol) and THF (30 ml) Medium methyl 2 (S) -O-trifuryl lactate (53 mg, 0.23 mmol) and 18-crown-6 (1 0 mg), and after completion of the reaction, after chromatography, yellow solid (60 mg) To give the title compound. FAB MS: 946 (M + Na)⁺And 902 (initial material + Na)⁺[Contaminants]. 6.43. Dodecylcarbamoylmethyl 4-O- (3 ', 4', 6'-tri-O-acetyl-2'-acetamide-2 '-Deoxy-β-D-glucopyranosyl) -6-O-benzoyl-3-O- [1 (R)-(carboxy) Ethyl] -2-deoxy-2-acetamide-α-D-glucopyranoside (43)   Compound (42) (50 mg) was treated with excess pyridine (1 ml) and acetic anhydride (0.5 ml) and DMAP (1 ml). 0 mg) for 2 hours at room temperature. The reaction mixture was dichloromethane (20m l), washed three times with 2N HCl (3x20 ml), dried over sodium sulfate, Evaporate to body (50 mg). The solid is dried under high vacuum overnight before the compound Hydrolyzed as in (42) and not purified as an oil (35 mg) Obtain thing (43). 6.44. Carboxymethyl 4,6-O-benzylidene-3-O- [methoxycarbonylmethyl] -2-deo Xy-2-acetamide-α-D-glucopyranoside (49)   Compound (14) was treated in the same manner as compound (10) with sodium periodate and Treated with um (III) chloride. The compound (49) (73%) is obtained by column purification. 6.45. Dodecylcarbamoylmethyl 4,6-O-benzylidene-3-O- [methoxycarbonylmethyl Ru] -2-deoxy-2-acetamide-α-D-glucopyranoside (49)   Compound (49) is 1-hydroxybenzotriazole, as in compound (11), 1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide meso-p-toluene Treatment with sulfonic acid and dodecylamine gives compound (50) (50%). 6.46. Dodecylcarbamoylmethyl 4,6-O-benzylidene-3-O- [carboxymethyl] -2-de Oxy-2-acetamide-α-D-glucopyranoside (51)   Compound (50) is dissolved in 1,4-dioxane as in the case of preparing compound (12), Treated with 0.1 M potassium hydroxide aqueous solution. Compound (51) is recovered in a quantitative yield Is done.6.47. α, β-D-Mannose pentaacetate (DL1)   D-Mannose (75 g, 0.41 mol) and 4-N, N dimethylamino in anhydrous pyridine (700 ml) One drop of acetic anhydride (300 ml) was added to a solution of norpyridine (5 g) stirred at 0 ° C. Are added one by one. The resulting solution is stirred at room temperature for 16 hours. This solvent Is removed by evaporation. The residue was dissolved in ethyl acetate (500 ml). And washed with 1M aqueous HCl to pH2. Organic layer is saturated NaHCO_ThreeWashed to pH 7 with , Separated and dried (Na_TwoSO_Four), Concentrated to 160 g of DLl (100%) as a white solid Get TLC (ethyl acetate: hexane 2: 3) Rf 0.34. 6.48. Phenyl 2,3,4,6-O-pentaacetyl-1-thio-D-mannopyranoside (DL2)   DLl (160 g, 0.41 mol) and thiophenol (49 g) in methylene chloride (1 L) at 0 ° C. , For a stirred solution of 0.45 mol, boron trifluoride diethyl etherate (116 g, 0.82 mol ) Is added drop by drop. The resulting solution is stirred at room temperature for 18 hours. Anti The reaction mixture is saturated Na_TwoCO_ThreeTo pH7. The organic layer was separated, dried (Na_TwoS 0_Four), To obtain 180g of DL2 (100%) as a thickened oil. TLC Cyl acetate: hexane 2: 3) Rf 0.42.6.49. Phenylthio-α-D-mannopyranoside (DL3)   To a stirred solution of DL2 (180 g, 0.41 mol) in anhydrous methanol (1 L) at 0 ° C, add Lithium methoxide (33 g, 0.61 mol) is added. The reaction mixture is stirred at room temperature for 16 hours Is done. For milky suspensions, Amberlite IR120 (plus) ion exchange Is added until the pH of the suspension is 6. The mixture is filtered. The filtrate is concentrated And co-evaporated with anhydrous toluene (5 × 50 ml). Residue is a small amount of methanol Dissolved in water. Etch until a white solid precipitates against the methanol solution. Ruacetate is added. The solid was collected by filtration to give 63 g of DL3 (56%). It is. The filtrate was concentrated to give oily DL3 (45 g, 40%), its TLC and¹H  NMR is identical to that of the solid. TLC (methanol: methylene chloride 1: 4) Rf0.63 . 6.50. Phenyl 4,6-O-benzylidene-1-thio-α-D-mannopyranoside (DL4)   DL3 (3.5 g, 12.8 mmol) and benzylidene dimethyl acetal (2.1 in DMF (40 ml)) To a stirred solution of 5 g (14 mmol) toluene sulfonic acid (300 mg) is added. this The solution is stirred at room temperature for 16 hours. The solvent is removed by evaporation. Residue Is dissolved in ethyl acetate and saturated Na_TwoC0_ThreeAnd then dried (Na_TwoS0_Four). solvent Is evaporated to give 3 g (67%) of DL4 as a solid. TLC (Methanol: Methyle Chloride 1: 9) Rf 0.51.6.51. Phenyl 2-O-benzyl-4,6-O-benzylidene-1-thio-α-D-mannopyranoside (DL5 )   Room temperature of DL4 (5.66 g, 16 mmol) and benzyl bromide (3.80 g, 22.4 mmol) in DMF (50 ml) Silver oxide (5.5 g) is added to the stirred solution. The suspension is stirred for 48 hours at room temperature . The reaction mixture is filtered. The solvent is removed by evaporation. The residue is vinegar Dissolved in ethyl acetate (50 ml), washed with water, dried (Na_TwoSO_Four), Concentrated. Shi Flash chromatography of the residue on silica (15% in hexane) Ethyl acetate) gives 3.6 g (50%) of DL5 as a thick oil. TLC (acetic acid Ethyl: hexane 1: 4) Rf 0.3. 6.52. Phenyl 3-O-allyl-2-O-benzyl-4,6-O-benzylidene-1-thio-α-D-mannopi Lanoside (DL6)   To a stirred solution of DL5 (3.5 g, 7.7 mmol) in anhydrous DMF (50 ml) at 0 ° C. was added NaH (95%, 390 mg) are added. The suspension is stirred for 30 minutes. Allyl bromide (1.86 g, 15.5 mmol ) Is added. The mixture is stirred at room temperature for 18 hours. Until a clear solution is observed At which methanol is added. The solvent is removed by evaporation. The residue is Acetic acid Dissolved in ethyl and saturated NH_FourWashed with Cl, dried (Na_TwoS0_Four), Concentrated mud 3.6 g (95%) of DL6 is obtained as a thickened oil. TLC (ethyl acetate: hexane 1: 9 ) Rf 0.41. 6.53. Phenyl 3-O-allyl-2,6-di-O-benzyl-1-thio-α-D-mannopyranoside (DL7)   DL6 in anhydrous THF (3 g, 6.1 mmol), NaBH_ThreeCN (3.7 g, 17 mmol) and molecular sieve 3Å (50 (0 mg) at 0 ° C, the HCl ether solution (1M) showed no air bubbles. Until it is no longer needed. The mixture is kept at 0 ° C. until TLC shows no initial material For 3 hours. The reaction mixture is poured into ice water. The mixture was ethyl acetate (3x30ml ), Dried and concentrated. The residue was purified by flash chromatography 2.55 g (85%) of DL as a waxy solid purified by (20% ethyl acetate in sun) 7 is obtained. TLC (ethyl acetate: hexane 1: 4) Rf 0.3. 6.54. Phenyl 4-O-acetyl-3-O-allyl-2,6-di-O-benzyl-1-thio-α-D-mannopira Noside (DL8)   Dispersion of DL7 (10 g, 20 mmol) and 4-N, N-dimethylpyridine (500 mg) in anhydrous pyridine Acetic anhydride (3.1 g, 30 mmol) is added to the stirred solution. The mixture is stirred at room temperature for 16 hours. Stirring Is done. The solvent is removed by evaporation. The residue is dissolved in ethyl acetate , Ice-cold HCl aqueous solution (1M), saturated NaHCO_ThreeAnd dried (Na_TwoSO_Four), Concentrated As a result, 11g (100%) of DL8 is obtained as a thick oil. TLC (ethyl acetate: Hexane 1: 2) Rf 0.73.6.55. Phenyl 4-O-acetyl-3-O-allyl-2,6-di-O-benzyl-1-thio-α-D-mannopira Noside S-oxide (DL9)   To a stirred solution of DL8 (11 g, 20 mmol) in ethyl acetate (100 ml) at −30 ° C. was added ethyl acetate. MCPBA (68.7%, 5.17 g, 20 mmol) in toluene (20 ml) is added dropwise. This solution is TLC Stir at -30 ° C for 30 minutes until indicates no initial material. Cold solution Saturated Na_TwoCO_ThreeWashed with. The organic layer is dried (Na_TwoSO_Four), Concentrated, muddy As a result, 11.5 g (100%) of DL9 is obtained as an oil. TLC (ethyl acetate: hexane 1: 2). 6.56. Methoxycarbonylmethylenyl 4-O-acetyl-3-O-allyl-2,6-di-O-benzyl- α-D-mannopyranoside (DL10) and methoxycarbonylmethylenyl 4-O-ace Tyl-3-O-allyl-2,6-di-O-benzyl-β-D-mannopyranoside (DL11)   To a stirred solution of DL9 (10.6 g, 19.3 mmol) in anhydrous toluene (200 ml) at -78 ° C, Fluoromethanesulfonic anhydride (5.4 g, 19.3 mmol) is added. 30 minutes stirring Afterwards, 2,6-di-t-butyl-4-methylpyridine in toluene (20 ml) is added. mixture Is Stir for 20 minutes. Methyl glycolate (1.8 g, 20 mmol) is added. The solution is -7 Stir at 8 to -70 ° C for 2 hours. Reaction is saturated Na_TwoCO_ThreeStopped with solution (100 ml) and acetic acid Extracted with ethyl (3x100ml), dried (Na_TwoSO_Four), Concentrated. The residue is flash Purified by liquid chromatography (10 → 25% ethyl acetate in hexane) 4 g (41%) of DL10 and 2 g (20.5%) of DL11 are obtained as grated oil. DL10: TLC (Ethyl acetate: hexane 1: 2) Rf 0.43. DL11: TLC (ethyl acetate: hexane 1: 2) Rf 0.31. 6.57. Methoxycarbonylmethylenyl 3-O-allyl-2,6-di-O-benzyl-α-D-mannopi Lanoside (DL12)   For a stirred solution of DL10 (4.0 g, 7.78 mmol) in anhydrous methanol (150 ml) at room temperature, Sodium methoxide (420 mg, 7.78 mmol) is added. The solution is stirred for 3 hours . Amberlite R120 (plus) ion exchange resin is added until the solution pH reaches 7. Is done. The mixture is then filtered and concentrated. The residue was dissolved in ethyl acetate (50 ml). H_TwoWashed with O, dried (Na_TwoSO_Four), Concentrated. Residue is flash chroma Mulled oil purified by chromatography (30% ethyl acetate in hexane) As a result, 2.5 g (68%) of DL12 is obtained. TLC (ethyl acetate: hexane 1: 2) Rf 0.25. 6.58. Phenyl 2,3,4,6-tetra-O-pivaloyl-1-thio-α-D-mannopyranoside S-oxo Sid (DL121)   D-glucose (20 g, 111 mmol) in pyridine (150 ml), trimethylacetyl chloride (80 g, 666 mmol) and DMAP (400 mg) are heated at 80 ° C. for 48 hours. Dissolution The medium is removed by evaporation. The residue was dissolved in ethyl acetate (150 ml), p Wash with cold aqueous HCl (1 M) until H2, add saturated NaHC0_ThreeWashed and dried ( Na_TwoSO_Four), Concentrated. Residue (70g) and thiophenol (13ml, 130mmol) Dissolved in lenchloride (200 ml). BF for the solution cooled in this ice bath_Three・ Et_TwoO ( 34 g, 240 mmol) are added dropwise. The mixture is stirred at room temperature for 16 hours, Sum NaHCO_ThreeNeutralized by The organic layer is dried (Na_TwoSO_Four), Concentrated. Residue (10 g, 16 .4 mmol) is dissolved in ethyl acetate (100 ml) and cooled to -30 ° C. Ethyl acetate (30ml MCPBA (4.1 g, 16.4 mmol) is added dropwise. The mixture is stirred at -30 ° C for 30 minutes And saturated Na_TwoCO_ThreeAt pH 8 and dried (Na_TwoSO_Four), Concentrated. The residue is Purified by flash chromatography (20% ethyl acetate in hexane) 8g of DL121 is obtained as a body. TLC (30% ethyl acetate in hexane) Rf 0.45. 6.59. Methoxycarbonylmethylenyl 3-O-allyl-2,6-di-O-benzyl-4-O- (2 ′, 3 ′, 4 ′ , 6'-Tetra-O-pivaloyl-1-β-D-glucosyl) -α-D-mannopyranoside (DL13)   Phenyl 2,3,4,6-tetra-O-pivaloyl-1-in ethyl acetate / hexane (1: 160 ml) Thio-α-D-mannopyranoside S-oxide (DL121, 6.9 g, 11 mmol) and 2,6-di-te To a stirred solution of rt-butyl-4-methylpyridine (2.25 g, 11 mmol) at -70 ° C, Fluoromethanesulfonic anhydride (3.1 g, 11 Eol) is added. Solution at -70 ° C 30 Stir for a minute. DL12 in ethyl acetate / hexane (1: 1.15 ml) is added. this The solution was stirred at -65 ° C for 2.5 hours, and saturated NaHCO_Three(30 ml) stops the reaction. Organic layer Are separated. The aqueous layer is extracted with ethyl acetate (3x30ml). This extract combines And dried (Na_TwoS0_Four), Concentrated and flash chromatography (hexane 5.0 g (93%) of DL13 as a thick oil Is obtained. TLC (ethyl acetate: hexane 1: 2) Rf 0.50. 6.60. Hydroxycarbonylmethylenyl 3-O-allyl-2,6-di-O-benzyl-4-O- (2 ′, 3 ′, 4 ', 6'-Tetra-O-pivaloyl-1-β-D-glucosyl) -α-D-mannopyranoside (DL 14)   A solution of DL13 (5 g, 5.15 mmol) in aqueous KOH (0.1 M) / dioxane (1: 1, 60 ml) at room temperature Stir for 24 hours. Amberlite IR120 (plus) ion exchange resin is the solution pH Until 6 is reached. The mixture was filtered, concentrated and dried in toluene (3x 20 ml) and 4.7 g (95%) of DL14 obtained as a thick oil. It is. TLC (methanol: methylene chloride 1: 9) Rf 0.5.6.61. Dodecylcarbamoylmethylenyl 3-O-allyl-2,6-di-O-benzyl-4-O- (2 ′, 3 ′, 4 ', 6'-Tetra-O-pivaloyl-1-β-D-glucosyl) -α-D-mannopyranoside (DL 15)   Compound DL14 (2 g, 2.1 mmol) in methylene chloride (30 ml), DCC (650 mg, 3.15 mmol) A mixture of HOBT (425 mg, 3.15 mmol) and dodecylamine (583 mg, 3.15 mmol) Stir at warm for 16 hours. The mixture is filtered and concentrated. The residue is flash black A muddy oil purified by chromatography (30% ethyl acetate in hexane) As a result, 2 g (84%) of DL15 is obtained. TLC (ethyl acetate: hexane 1: 2) Rf 0.3. 6.62. Dodecylcarbamoylmethylenyl 2,6-di-O-benzyl-4-O- (2 ', 3', 4 ', 6'-tetra La-O-pivaloyl-1-β-D-glucosyl) -α-D-mannopyranoside (DL16)   PdCl to a solution of DL15 (800 mg, 0.7 mmol) in methanol (25 ml)_Two(64mg) is added You. The suspension may be allowed to stand at room temperature for 2.5 hours or Stir until indicated. The suspension is filtered. The filtrate is concentrated and short silica Purified on a column (35% ethyl acetate in hexane), slushy 720 mg (93%) DL16 Obtained as oil. TLC (35% ethyl acetate in hexane) Rf 0.45.6.63. Dodecylcarbamoylmethylenyl 2,6-di-O-benzyl-3-O- [1 ′ (R)-(carboxy ) Ethyl] -4-O- (2 ', 3', 4 ', 6'-tetra-O-pivaloyl-1-β-D-glucosyl) -α-D- Mannopyranoside (DL17)   To a stirred solution of DL16 (720 mg, 0.66 mmol) in dry THF was added sodium hydride (9 5%, 66 mg, 2.6 mmol) are added. The suspension is stirred for 30 minutes. Reagent (S)-(-)-2 Bromopropionic acid (150 mg, 1 mmol) is added. The mixture is stirred for 16 hours. Methanol (2 ml) is added. The solution is evaporated. The residue is dissolved in ethyl acetate Disassembled and washed with aqueous HCl (1M). The organic layer is dried (Na_TwoS0_Four), Concentrated, Purified by flash chromatography (6-8% methanol in methylene chloride) 500 mg (65%) of DL17 is obtained as a thick oil. TLC 10% methanol in chloride) Rf 0.45. 6.64. Dodecylcarbamoylmethylenyl 3-O- [1 '(R)-(carboxy) ethyl] -4-O- (2', 3 ', 4', 6'-Tetra-O-pivaloyl-1-β-D-glucosyl) -α-D-mannopyranoside (D L18)   DL17 (500 mg, 0.5 mmol) and palladium on carbon (10%, 300 m2) in ethanol (10 ml) The suspension of g) is subjected to hydrogenolysis at 50 psi for 48 hours. The suspension is filtered and concentrated , 400 mg (95%) of DL18 are obtained as a thick oil. 6.65. Dodecylcarbamoylmethylenyl 2,6-O-acetyl-3-O- [1 '(R)-(carboxy) Tyl] -4-O- (2 ', 3', 4 ', 6'-tetra-O-pivaloyl-1-β-D-glucosyl) -α-D- Mannopyranoside (DL19)   DL18 (340 mg, 0.35 mmol), acetic anhydride (107 mg, 3.15 mmol) and D in anhydrous pyridine A solution of MAP (10 mg) is stirred at room temperature for 16 hours. This solution is evaporated. Residue Is dissolved in ethyl acetate (10 ml) and added to an aqueous HCl solution (2M, 5 ml). The mixture is in the chamber Stir at warm for 2 hours. The organic layer is dried (Na_TwoS0_Four), Concentrated, flash Purified by chromatography (2-10% methanol in methylene chloride), 55 mg (17 %) DL19 is obtained as a thick oil. TLC (10% medium in methylene chloride (Tanol) Rf 0.65. 6.66. Phenyl 2-deoxy-2-N-phthalimido-1,3,4,6-tetra-O-acetyl-α-D-gluco Pyranoside (RC1)   For a suspension of D-glucosamine hydrochloride (30 g, 138.9 mmol) in methanol (300 ml) Amberlite RA-400 (OH) ion exchange resin (99.2 ml, 138.9 mmol) was added. To . The reaction mixture is stirred at room temperature for 1 hour. The resin is removed by filtration and the filtrate Is treated with phthalic anhydride (20.6 g, 138.9 mmol). Reaction mixture at room temperature for 1.5 hours Stir and the solid intermediate product is collected by filtration and dried. Phthalic anhydride (2 (0.6 g, 138.9 mmol) is added to the filtrate and the reaction mixture is stirred overnight at room temperature. Sa Furthermore, a solid intermediate product is obtained. The two parts of the intermediate product are mixed ( 29.5g, 64%). Next, 6 g (21.3 mmol) of the intermediate product was dissolved in dichloromethane (70 ml). Is done. Pyridine (26 ml, 319.5 mmol), acetic anhydride (20 ml, 213 mmol), and DMAP (0 .4g, 15%) is added. Stir at room temperature overnight, then add dichloromethane (100ml) Is done. The reaction solution was continuously 1N HCl (2 × 100 ml), saturated aqueous sodium bicarbonate (2 × 1 00ml), water (2x100ml), and brine (2x100ml), dried (Na_TwoSO_Four), Dark Contracted. The crude product (Qy) is used in the next step without further purification. TLC (Et OAc-hexane 1: 1) Rf 0.35. 6.67. Phenyl 2-deoxy-2-N-phthalimido-1-thio-3,4,6-tri-O-acetyl-α-D-glu Copyranoside (RC2)   To a cooled (0 ° C.) solution of RCl (32 g, 67 mmol) in anhydrous dichloromethane (200 ml) Thiophenol (7.6 ml, 73.7 mmol) was added, followed by boron trifluoride Chilletherate (16.5 ml, 134 mmol) is added. Reaction mixture is warmed to room temperature And stirred at 50 ° C. overnight. Dichloromethane (100 ml) was added You. The reaction solution was continuously saturated aqueous sodium bicarbonate (2x200ml), water (2x200ml) , And brine (2x200 ml), dried (Na_TwoSO_Four), Concentrated. As a result The resulting product RC2 (Qy) is used in the next step without further purification. TLC (E tOAc-hexane 1: 1) Rf 0.45. 6.68. Phenyl 2-deoxy-2-N-phthalimide-1-thio-β-D-glucopyranoside (RC3)   To a solution of RC2 (34.0 g, 64.5 mmol) in methanol (200 ml) was added sodium methoxide Cid (3.5 g, 64.8 mmol) is added. The reaction mixture is stirred overnight at room temperature. Ann Burlite IR-120 plus (H) ion exchange resin (50 ml, 95 mmol) is added. Mixed The mixture is stirred for 1 hour. The resin was removed by filtration and the filtrate was dried (MgSO_Four ), Concentrated. The resulting product, RC3 (Qy), can be used without further purification. Used in the stage. TLC (EtOAc-hexane 7: 3) Rf 0.42. 6.69. Phenyl 2-deoxy-4,6-isopropylidene-2-N-phthalimide-1-thio-β-D-glu Copyranoside (RC4)   P-top against a solution of RC3 (54.0 g, 134.6 mmol) in 2,2-dimethoxypropane (200 ml) Ruenesulfonic acid (2.6 g, 13.46 mmol) is added. The reaction mixture is stirred overnight at room temperature. Stir. Ethyl acetate (200ml) is added. The reaction mixture is saturated sodium bicarbonate Water (2x300ml), water (2x300ml), and brine (2x300ml), dried ( Na_TwoSO_Four), Concentrated. The resulting product RC4 (Qy) requires no further purification. Used in the next stage. TLC (EtOAc-hexane 1: 1) Rf 0.6. 6.70. Phenyl 2-deoxy-4,6-isopropylidine-3-O-p-methoxybenzyl-2-N-lid Limid-1-thio-β-D-glucopyranoside (RC5)   A solution of RC4 (60 g, 136 mmol) in tetrahydrofuran (200 ml) was treated with sodium hydride (6.87 g, 272 mmol) / tetrahydrofuran (30 ml). It is. p-Methoxybenzyl chloride (27.66 ml, 204 mmol) is added. Reaction mixing The solution is stirred at 65 ° C. for 24 hours. The reaction was quenched with 2N HCl (200 ml) and ethyl acetate (3 00 ml) is then added. The organic layer was separated, water (2x300ml), and brine (2x300ml) ml), dried (Na_TwoSO_Four), Concentrated. Residue is flash chromatography Raffy (EtOAc-hexane 3: 7, 2: 3, 1: 1) and purified as a viscous liquid with RC5 (35 g, 63%) and RC4 (16.2 g) are obtained. TLC (ethyl acetate: hexane 3: 7) Rf 0.4. TLC (ethyl acetate: hexane 1: 1) Rf 0.5. 6.71. Phenyl 2-deoxy-3-O-p-methoxybenzyl-2-N-phthalimide-1-thio-β-D-glu Copyranoside (RC6)   To a solution of RC5 (3.6 g, 6.42 mmol) in methanol (36 ml) was added toluenesulfonic acid ( 0.12 g, 0.642 mmol) are added. The reaction mixture is stirred at room temperature for 4.5 hours. Ann Barlite RA-400 (OH) ion exchange resin (4.6 ml, 6.44 mmol) was added and the mixture Is stirred for 1 hour. The resin was removed by filtration and the filtrate was dried (Na_TwoSO_Four) , Concentrated. The resulting product RC6 (Qy) can be used without further purification in the next step. Used in stages. TLC (EtOAc-hexane 7: 3) Rf 0.2. 6.72. Phenyl 6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl-2-N-phthalimido-1- Thio-β-D-glucopyranoside (RC7)   A solution of RC6 (7.39 g, 14.2 mmol) in toluene (100 ml) is heated under reflux for 1 hour. The Butyl tin oxide (3.45 g, 14.2 mmol) is added. The reaction mixture was added under reflux overnight. Get heated. Tetrabutylammonium iodide (5.24 g, 14.2 mmol) and bromide bromide Anzil (2.2 ml, 18.5 mmol) is added to the reaction mixture and stirred at 80 ° C. for 32 hours. Ethyl acetate (150 ml) is added. The reaction mixture was water (2x150 ml), and brine (2x150 ml). ml), dried (Na_TwoSO_Four), Concentrated. Residue is flash chromatography Raffy (EtOAc: Hexanes 3: 7 to 3: 2) was purified and RC7 (6 g , 69%). TLC (EtOAc: hexane 1: 1) Rf 0.45. 6.73. Phenyl 2-deoxy-2-N-phthalimido-1-thio-3,4,6, -tri-O-benzyl-β-D-g Lucopyranoside (RC8)   A solution of RC3 (22.5 g, 56.1 mmol) in tetrahydrofuran (150 ml) was prepared (8.5 g, 336.6 mmol) / tetrahydrofuran (20 ml) suspension. Bromination Benzyl (24 ml, 202 mmol) is added. The reaction mixture is stirred overnight at 65 ° C. The reaction is quenched with 2N HCl (100 ml) and ethyl acetate (200 ml) is added. Organic layer is water (2x200ml), and brine (2x200ml), dried (Na_TwoSO_Four), Concentrated. The residue was purified by flash chromatography (EtOAc: hexane 1: 9, 1: 4, 3: 7). To give RC8 (17.42 g, 46%) as a viscous liquid. TLC (ethyl acetate: Sun 3: 7) Rf 0.55. 6.74. Dodecyl 2-deoxy-2-N-phthalimide-3,4,6, -tri-O-acetyl-β-D-glucopyr Noside (RC9)   RC1 (9.85 g, 20.6 mmol), 1-dodecanol (3.8 g, 20. 6 mmol) and a stirred mixture of molecular sieves. (10 ml, 51.5 mmol) is added. The reaction mixture is stirred at room temperature for 3 hours. reaction The mixture was poured into saturated aqueous sodium bicarbonate (100 ml) and ethyl acetate was added. You. The organic layer was separated, washed with water (2x150ml), and brine (2x150 (Na_TwoSO_Four), Concentrated. The residue was purified by flash chromatography (EtOAc: Purification by Sun 3: 7) gives RC9 (7.83 g, 63%) as a white solid. TLC (E tOAC-hexane 3: 7) Rf 0.55. 6.75. Dodecyl 2-deoxy-2-N-phthalimide-β-D-glucopyranoside (RC10)   To a solution of RC9 (10.35 g, 17.1 mmol) in methanol (50 ml) was added sodium methoxide. Cid (0.93 g, 17.1 mmol) is added. The reaction mixture is stirred at room temperature for 3 hours. A Amberlite IR-120 plus (H) ion exchange resin (12 ml, 2.04 mmol) is added . The reaction mixture is stirred for one hour. Resin is removed by filtration and the filtrate is dried ( MgSO_Four), Concentrated. The resulting product, RC10 (Qy), requires no further purification. Used in the next stage. TLC (EtOAc) Rf 0.60. 6.76. Dodecyl 2-deoxy-4,6-isopropylidene-2-N-phthalimido-β-D-glucopyrano Sid (RC11)   For a solution of RC10 (5.7 g, 11.95 mmol) in 2,2-dimethoxypropane (50 ml), p-top Ruenesulfonic acid (285 mg, 15%) is added. The reaction mixture is stirred at room temperature for 5 hours. It is. Ethyl acetate (100 ml) is added. The organic layer is separated and saturated sodium bicarbonate Aqueous (2 x 150 ml), water (2 x 150 ml), and brine (2 x 150 ml) and dried. (Na_TwoSO_Four), Concentrated. The residue was purified by flash chromatography (EtOAc: 3:11 to 3: 7) to give RC11 (2.02 g, 39%) as a viscous liquid . TLC (EtOAc: hexane 1: 1) Rf 0.6. 6.77. Dodecyl 2-deoxy-4,6-isopropylidine-3-O-p-methoxybenzyl-2-N-lid Limid-β-D-glucopyranoside (RC12)   A solution of RC11 (2.0 g, 3.87 mmol) in tetrahydrofuran (12 ml) was treated with sodium hydride. (147 mg, 5.81 mmol) / tetrahydrofuran (3 ml) was added dropwise. You. p-Methoxybenzyl chloride (0.79 ml, 5.81 mmol) is added. Reaction mixture Is Stir at 65 ° C for 24 hours. The reaction was stopped with 2N HCl (20 ml) and ethyl acetate (50 ml) was added. Is added. The organic layer was separated and washed with water (2x100ml), and brine (2x100ml) , Dried (Na_TwoSO_Four), Concentrated. The residue was purified by flash chromatography (EtOAc : Hexane: 3: 7) to obtain RC12 (1.28 g, 63%) as a viscous liquid . TLC (EtOAc: hexane 3: 7) Rf 0.6. 6.78. Dodecyl 2-acetamido-2-deoxy-4,6-isopropylidine-3-O-p-methoxyben Jil-β-D-glucopyranoside (RC13)   RC13 (623 mg, 1 mmol) and hydrazine hydrate (1 ml, 20 mmol) in 95% EtOH (10 ml) The solution is heated under reflux for 6.5 hours. The white precipitate was removed by filtration and the filtrate was concentrated. Contracted. The residue is dissolved in pyridine (10 ml) and acetic anhydride (1.4 ml, 15 mmol) . The reaction mixture is stirred at room temperature for 2 hours. Methanol is added and the reaction mixture is Stir at room temperature for 1 hour. Ethyl acetate was added (100 ml) and the reaction mixture was saturated Wash with aqueous sodium acid solution (2x50ml), water (2x50ml), and brine (2x50ml) and dry Dried (Na_TwoSO_Four), Concentrated. The residue was purified by flash chromatography (EtOAc- (13:17 to 3: 7) to afford RC13 (320 mg, 60%) as a viscous liquid. It is. TLC (EtOAC: hexane 3: 7) Rf 0.6. TLC (EtOAc: hexane 1: 1) Rf 0.25. 6.79. Dodecyl 2-acetamide-2-deoxy-3,4-isopropylidene-β-D-glucopyranosi C (RC14)   To a solution of RC13 (300 mg, 0.56 mmol) in dichloromethane (10 ml) was 2,3-dichloromethane. B-5,6-dicyano-1,4-benzoquinone (165 mg, 0.73 mmol) is added. Reaction mixing The solution is stirred at room temperature for 1.5 hours. The reaction mixture is a 10% sodium bisulfite solution (30 ml) And dichloromethane (50 ml) is added. The organic layer was separated and water (2x50ml) , And brine (2x30 ml), dried (Na_TwoSO_Four), Concentrated. The residue is hula Purified by flash chromatography (EtOAc-MeOH 1: 0 to 9: 1), viscous liquid RC14 (180 mg, 75%) is obtained as a body.6.80. Dodecyl 2-acetamido-3-O- (1-carboxy) ethyl-2-deoxy-4,6-isopropyl Lysine-β-D-glucopyranoside (RC15)   For a stirred solution of RC14 (180 mg, 0.42 mmol) in anhydrous tetrahydrofuran (4 ml), Sodium hydride (42.4 mg, 1.68 mmol) is added. Reaction mixture for 30 minutes at room temperature Stirred. (S)-(-)-2 Bromopropionic acid (0.057 ml, 0.63 mmol) was added and 24 Stir at room temperature for hours. The reaction mixture was poured into HCl (2N, 20 ml) and ethyl acetate (50 ml ) Is added. The organic layer was separated and washed with water (2x50ml), and brine (2x30ml) And dried (Na_TwoSO_Four), Concentrated. Flash chromatography (EtOAc-Me OH 1: 0 to 9: 1) to give RC15 (140 mg, 67%) as a viscous liquid . 6.81. Phenyl 6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl-2-N-phthalimido-1- Thio-4-O-trimethylsilyl-β-D-glucopyranoside (TYC2)   TYCl (5.94 g, 9.72 mmol) and triethylamine (2. 7 ml (19.4 mmol) of a cooled (-70 ° C) solution was added to trimethylsilyl trifluorofluoride. Lomethanesulfonate (2.3 ml, 11.9 mmol) is added. Reaction mixture for 1 hour Stir at 70 ° C. The reaction mixture is poured into a saturated aqueous sodium bicarbonate solution (50 ml) . The organic layer is separated and the aqueous layer is extracted with dichloromethane (50ml). Matched The organic layer was washed with water (100 ml), and saturated brine (50 ml), dried (Na_TwoSO_Four), Dark Contracted. The crude product RC2 (quantitative yield, or Qy) can be used without further purification Used in stages. TLC (EtOAc-hexane 1: 1) Rf 0.66. 6.82. Phenyl 6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl-2-N-phthalimido-1- Sulfinyl-4-O-trimethylsilyl-β-D-glucopyranoside (TYC3)   To a cooled (-60 ° C) solution of TYC2 (0.54 g, 0.79 mmol) in ethyl acetate (5.Oml) And 3-chloroperoxybenzoic acid (68.7%, 0.20 g, 0. 79 mmol) solution is added. Allow the reaction mixture to slowly warm to -30 ° C in one hour. Is placed. The reaction mixture was poured into a saturated aqueous sodium bicarbonate solution (20 ml), (20 ml) is added. The organic layer was separated and the aqueous layer was extracted with dichloromethane (50ml) Is done. The combined organic layers were washed with water (20 ml) and saturated brine (20 ml), and After drying (Na_TwoSO_Four), Concentrated. The crude product (Qy) can be Used in stages. TLC (EtOAc-hexane 1: 1) Rf 0.48. 6.83. Dodecyl 6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl-2-N-phthalimido-β -D-glucopyranoside (TYC4)   TYC3 (2.15 g, 3.07 mmol), 2,6-di-tert-butyl in EtOAc-hexane (1: 1 v / v 30 ml) 4-Methylpyridine (0.63 g, 3.07 mmol) and 1-dodecanol (0.57 g, 3.07 mmol) ) Solution was dropped into an ice bath (-40 ° C), and then Sulfonic anhydride (0.56 ml, 4.65 mmol) is added. Reaction mixture at -40 ° C for 30 minutes While stirring. The reaction mixture is poured into a saturated aqueous sodium bicarbonate solution (30 ml). Yes The organic layer is separated, and the aqueous layer is extracted with ethyl acetate (30 ml). The combined organic layers Washed with water (60 ml), and saturated brine (50 ml), then dried (Na_TwoSO_Four), Concentrated It is. The residue was purified by flash chromatography (EtOAc-hexane 3: 7 to 10: 0). TYC4 (0.75 g, 38%) is obtained as a viscous liquid. TLC (EtOAc-hex Sun 3: 7) Rf 0.25. TLC (EtOAc-hexane 1: 1) Rf 0.65. 6.84. Phenyl 2-deoxy-2-N-phthalimido-1-sulfinyl 3,4,6-tri-O-benzyl-β -D-glucopyranoside (TYC6)   To a cooled (−60 ° C.) solution of TYC5 (0.28 g, 0.42 mmol) in ethyl acetate (3.0 ml), vinegar was added. 3-chloroperoxybenzoic acid (68.7%, 0.11 g, 0.43 mm) in ethyl acetate (1.0 ml) ol) solution is added. The reaction mixture is allowed to warm to -30 ° C in 1 hour . Anti The reaction mixture was poured into a saturated aqueous solution of sodium bicarbonate (20 ml), and ethyl acetate (30 ml) was added. Be added. The organic layer was separated, washed with water (20ml), and saturated brine (20ml), After drying (Na_TwoSO_Four), Concentrated. The crude product (Qy) can be Used in stages. TLC (EtOAc-hexane 1: 1) Rf 0.42. 6.85. Dodecyl (2-deoxy-2-N-phthalimide-3,4,6-tri-O-benzyl-β-D-glucopyr Nosyl)-(1 → 4) -2-deoxy-6-O-benzyl-3-O-p-methoxybenzyl-2-N-phthaly Mid-β-D-glucopyranoside (TYC7)   TYC4 (0.23 g, 0.33 mmol), TYC6 (0.53 g, 0.77 mm) in EtOAc-hexane (1: 1 v / v 6 ml) ol), and 2,6-di-tert-butyl-4-methylpyridine (0.16 g, 0.78 mmol) cooled (-6 (0 ° C) to the solution was added trifluoromethanesulfonic anhydride (0.093 ml, 0.77 mmol) Is done. The reaction mixture is stirred at -60 ° C for 30 minutes. The reaction mixture is saturated sodium bicarbonate Pour into aqueous lithium solution (20 ml) and add ethyl acetate (30 ml). Organic layer separated And the aqueous layer is extracted with ethyl acetate (30 ml). The combined organic layers are water (60 ml), And saturated brine (50 ml), then dried (Na_TwoSO_Four), Concentrated. The residue is Purified by flash chromatography (from EtOAc-hexane 1: 9) 5: 5, TYC7 (0.19 g, 48%) is obtained as a thick liquid. TLC (EtOAc-hexane 3: 7) Rf 0.19 . TLC (EtOAc-hexane 1: 1) Rf 0.76. 6.86. Dodecyl (2-acetamido-2-deoxy-3,4,6-tri-O-benzyl-β-D-glucopyrano (Sil)-(1 → 4) -2-acetamido-6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl -β-D-glucopyranoside (TYC8)   TYC7 (1.57 g, 1.26 mmol) and hydrazine hydration in 95% EtOH-THF (1: 1 v / v, 26 ml) (1.26 ml, 26.0 mmol) solution is heated under reflux for 24 hours. White precipitate is removed by filtration And the filtrate is concentrated. The residue was dissolved in pyridine (12 ml) and acetic anhydride (1.2 ml, 12.7 mmol) are added. The reaction mixture is stirred at room temperature for 2 hours. methanol (2 ml, 49.4 mmol) is added. The reaction mixture is stirred at room temperature for 1 hour. Reaction mixing The solution was poured into saturated aqueous sodium bicarbonate (50 ml) and ethyl acetate (100 ml) was added. It is. The organic layer was separated, washed with water (100 ml), and brine (50 ml), dried ( Na_TwoSO_Four), Concentrated. The residue was purified by flash chromatography (EtOAc-DCM 5: 5 7: 3) to give TYC8 (0.85 g, 63%) as a viscous liquid. TLC (Et OAc: DCM 3: 7) Rf 0.47. 6.87. Dodecyl (2-acetamido-2-deoxy-3,4,6-tri-O-benzyl-β-D-glucopyrano (Sil)-(1 → 4) -2-acetamido-6-O-benzyl-2-deoxy-β-D-glucopyranoside ( TYC9)   To a solution of TYC8 (0.85 g, 0.78 mmol) in 95% dichloromethane (15 ml) was 2,3-dic Loro-5,6-dicyano-1,4-benzoquinone (0.23 g, 1.01 mmol) is added. Reaction mixture The mixture is stirred at room temperature for 30 minutes. The reaction mixture was a 10% sodium bisulfite solution (30 ml ) And dichloromethane (30 ml) is added. The organic layer was separated and water (40ml) , And saturated brine (30 ml), dried (Na_TwoSO_Four), Concentrated. The residue is Purified by lash chromatography (EtOAc: DCM 5: 5 to 8: 2), viscous liquid TYC9 (0.56 g, 74%) is obtained as a body. TLC (EtOAc: DCM 7: 3) Rf 0.48.6.88. Dodecyl (2-acetamido-2-deoxy-3,4,6-tri-O-benzyl-β-D-glucopyrano (Sil)-(1 → 4) -2-acetamido-6-O-benzyl-3-O- (1-carboxy) ethyl-2-deoxy Si-β-D-glucopyranoside (TYC10)   To a solution of TYC9 (0.39 g, 0.41 mmol) in THF-DMF (1: 1 v / v, 16 ml) was added sodium hydride (39 mg, 1.63 mmol) is added. The reaction mixture is stirred at room temperature for 30 minutes. (S )-(-)-2Bromopropionic acid (0.055 ml, 0.61 mmol) is added. 24 hours at room temperature Stir. The reaction mixture is poured into 2M hydrochloric acid (20ml) and ethyl acetate (50ml) is added . The organic layer was separated, washed with water (40ml), and saturated brine (30ml), dried (N a_TwoSO_Four), Concentrated. Purified by flash chromatography (MeOH: DCM 1: 9) This gives TYC10 (0.26 g, 62%) as a viscous liquid. 6.89. Dodecyl (2-acetamido-2-deoxy-β-D-glucopyranosyl)-(1 → 4) -2-aceta Mido-3-O- (1-carboxy) ethyl-2-deoxy-β-D-glucopyranoside (TYC11)   For a solution of TYC10 (0.26 g, 0.25 mmol) in ethanol (100 ml), add 10% palladium on carbon. (0.50 g) is added. The reaction mixture was brought to room temperature in a Parr hydrogenator at 40 psi. g is shaken for 24 hours in the presence of hydrogen gas. The reaction mixture is filtered through filter paper, It is concentrated. The crude product (0.15 g, 89%) was used in the next step without further purification It is. 6.90. Dodecyl (2-acetamide-2-deoxy-3,4,6-tri-O-acetyl-β-D-glucopyrano (Sil)-(1 → 4) -2-acetamido-6-O-acetyl-3-O- (1-carboxy) ethyl-2-deoxy Si-β-D-glucopyranoside (TYC12)   For a solution of TYC11 (0.15 g, 0.23 mmol) in pyridine (10 ml), acetic anhydride (0.43 ml, 4 .56 mmol) are added. The reaction mixture is stirred at room temperature for 2 hours. Ethyl acetate (100m l) and 2M hydrochloric acid (100 ml) are added. The organic layer was separated, water (50 ml), and saturated Wash with brine (50 ml), dry (Na_TwoSO_Four), Concentrated. Crude product (0.16 g, 85% ) Is used in the next step without further purification. 6.91. Methyl (3α, 5β) -3-methoxycholane-24-oate (TYC24)   To a solution of lithocholic acid (31.40 g, 83.5 mmol) in dimethylformamide (165 ml) Then sodium hydride (6.01 g, 0.25 mol) is added. Reaction mixture at room temperature 30 Stir for a minute. Iodomethane (15.6 ml, 0.25 mol) is added. The reaction mixture Stir at room temperature for 3 hours. The reaction mixture was poured into 2M hydrochloric acid (100ml) and dissolved in ether-DCM. A medium mixture (2: 1 v / v, 500 ml) is added. The organic layer was separated and saturated brine (200 ml) And saturated aqueous potassium carbonate (200 ml), dried (Na2S04) and concentrated You. The crude product (Qy) is used in the next step without further purification. 6.92. (3α, 5β) -3-methoxycholan-24-ol (TYC15)   Cooling of TYC14 (37.43 g, 92.6 mmol) in THF-ether solvent mixture (2: 1 v / v, 450 ml) (0 ° C) solution, lithium aluminum hydride (7.04 g, 0.185 mol) was added. It is. The reaction mixture is stirred at 0 ° C. for 2 hours. The reaction mixture is 1M sodium hydroxide The addition is stopped by dropping the liquid. The reaction mixture was poured into 2M hydrochloric acid (200ml) , Diethyl ether (300 ml) is added. The organic layer was separated, water (200 ml), and And saturated brine (200 ml), dried (Na_TwoSO_Four), Concentrated. With solvent mixture Some EtOAc-hexane (3: 7 v / v, 400 ml) is added to the residue. The resulting white The color precipitate is removed by filtration. The filtrate is concentrated. Residue from methanol-water Crystallized to give TYC15 as a white solid. Crude product (17.26 g, 55%) is further purified Will be used in the next step without performing. TLC (EtOAc-hexane 1: 1) Rf 0.63. 6.93. (3α, 5β) -24- (6-O-benzyl-2-deoxy-3-O-p-methoxybenzyl-2-N-phthaly (Mido-β-D-glucopyranosyloxy) -3-methoxycholane (TYC16)   TYC15 (7.09 g, 18.8 mmol) in EtOAc-hexane solvent mixture (1: 1 v / v, 190 ml), TYC 3 (13.17 g, 18.8 mmol) and 2,6-di-tert-butyl-4-methylpyridine (3.86 g, 18.8 mmol) solution was added dropwise to the cooling bath (-40 ° C) and then quickly added to the trifluorometa Sulfonic anhydride (3.74 ml, 31.0 mmol) is added. Reaction mixture is -40 ° C for 30 minutes And stirred. The reaction mixture was poured into a saturated aqueous solution of sodium bicarbonate (100 ml). You. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (100ml). Combined The organic layer was washed with water (200 ml), and saturated brine (100 ml), dried (Na_TwoSO_Four),concentrated Is done. The residue is extracted with a solvent mixture of EtOAc-hexane (3: 7 v / v, 200 ml). This The filtrate of is concentrated. The residue was purified by flash chromatography (EtOAc-hexane 3 : 7 to 5: 5) to give TYC16 (2.54 g, 15%) as a viscous liquid. T LC (EtOAc-hexane 1: 1) Rf 0.66. 6.94 (3α, 5β) -24-[(2-deoxy-2-N-phthalimido-3,4,6-tri-O-benzyl-β-D-glu (Copyranosyl)-(1 → 4) -2-deoxy-6-O-benzyl-3-O-p-methoxybenzyl-2-N- Phthalimide-β-D-glucopyranosyloxy] -3-methoxycholane (TYC17) TYC16 (2.15 g, 2.45 mmol), TYC6 (4.66 g, 6.78) in EtOAc-hexane (1: 1 v / v, 50 ml) mmol) and 2,6-di-tert-butyl-4-methylpyridine (1.76 g, 8.59 mmol) cooled (-4 (0 ° C.), trifluoromethanesulfonic anhydride (0.82 ml, 6.81 mmol) was added to the solution. Added. The reaction mixture is stirred for 30 minutes at -40 ° C. The reaction mixture was sodium bicarbonate It is poured into a saturated aqueous solution of um (50 ml). The organic layer was separated and the aqueous layer was ethyl acetate (3 0ml). Wash the combined organic layers with water (60 ml) and saturated brine (50 ml) And dried (Na_TwoSO_Four), Concentrated. The residue was purified by flash chromatography (E Purified by tOAc-hexane 3: 7 to 5: 5). The resulting product (3.26g) Used in the next step without further purification. TLC (EtOAc: hexane 3: 7) Rf 0.2 Five. TLC (EtOAc: hexane 1: 1) Rf 0.60. Fab MS: 1462 (M + H + Na)⁺. 6.95. (3α, 5β) -24-[(2-acetamido-2-deoxy-3,4,6-tri-O-benzyl-β-D-gluco (Pyranosyl)-(1 → 4) -2-acetamido-6-O-benzyl-2-deoxy-3-O-p-methoxybe Ndyl-β-D-glucopyranosyloxy] -3-methoxycholane (TYC18)   TYC17 in 95% EtOH-THF (1: 1 v / v, 40 ml) solvent mixture (not purified, 3.26 g ) And hydrazine hydrate (4.39 ml, 90.6 mmol) solution are heated under reflux for 24 hours. The white precipitate is removed by filtration and the filtrate is concentrated. Residue in pyridine (20 ml) Dissolve and add acetic anhydride (4.27 ml, 45.3 mmol). Reaction mixture at room temperature for 2 hours While stirring. Methanol (3.67 ml, 90.7 mmol) is added. Reaction mixture at room temperature For 1 hour. The reaction mixture was poured into saturated aqueous sodium bicarbonate (50 ml). , Ethyl acetate (100 ml) is added. The organic layer was separated, water (50ml), and saturated Wash with brine (50 ml), dry (Na_TwoSO_Four), Concentrated. The residue is EtOAc-hexane Washing with a mixed solvent (6: 4 v / v) gives TYC18 as a white solid. Crude product (1.92g) Is used in the next step without further purification. Fab MS: 1286 (M + H + Na)⁺. 6.96. (3α, 5β) -24-[(2-acetamido-2-deoxy-3,4,6-tri-O-benzyl-β-D-gluco Pyranosyl)-(1 → 4) -2-acetamido-6-O-benzyl-2-deoxy-β-D-glucopyr Nosyloxy] -3-methoxycholane (TYC19)   For a solution of TYC18 (crude, 1.92 g) in 95% aqueous dichloromethane (30 ml), 2 , 3-Dichloro-5,6-dicyano-1,4benzoquinone (0.45 g, 1.98 mmol) is added. The reaction mixture is stirred at room temperature for 20 minutes. The reaction mixture is 10% sodium bisulfite Pour into the solution (30 ml) and add dichloromethane (30 ml). The organic layer is separated, Wash with water (40 ml), and saturated brine (30 ml), then dry (Na_TwoSO_Four), Concentrated It is. The residue was purified by flash chromatography (MeOH: DCM 5:95), TYC9 (0.62 g, 19% from TYC16) is obtained as a viscous liquid. TLC (MeOH: DCM 1: 9) Rf  0.50. 6.97. (3α, 5β) -24-[(2-acetamido-2-deoxy-3,4,6-tri-O-benzyl-β-D-gluco (Pyranosyl)-(1 → 4) -2-acetamido-6-O-benzyl-3-O- (1-carboxy) ethyl-2- Deoxy-β-D-glucopyranosyloxy] -3-methoxycholane (TYC20)   For a TYC19 solution in THF-DMF (2: 1 v / v, 9 ml), sodium hydride (68 mg, 2.83 mm ol) is added. (S)-(-)-2 Bromopropionic acid (0.064 ml, 0.71 mmol) was added. It is. The reaction mixture is stirred at room temperature for 24 hours. The reaction mixture was poured into 2M hydrochloric acid (20ml) And ethyl acetate (50 ml) is added. The organic layer was separated, water (40ml), and saturated Wash with brine (30 ml) then dry (Na_TwoSO_Four), Concentrated. Crude product (0.57g) Is used in the next step without further purification. TLC (MeOH: DCM 1: 9) Rf 0.25. Fab MS: 1260 (M + Na + Na)⁺. 6.98. (3α, 5β) -24-[(2-acetamide-2-deoxy-β-D-glucopyranosyl)-(1 → 4) -2- Acetamide-3-O- (1-carboxy) ethyl-2-deoxy-β-D-glucopyranosyloxy [S] -3-methoxycholane (TYC21)   For a solution of TYC20 (unpurified, 0.57 g) in ethanol (100 ml), add 10% palladium on carbon. (0.57 g) is added. The reaction mixture was placed in a Parr hydrogenator at room temperature Shake with sig in the presence of hydrogen gas for 24 hours. The reaction mixture is filtered through filter paper, It is concentrated. The crude product (0.39 g, 96% from TYC19) was used in the next step without further purification. Used on floors. 6.99. (3α, 5β) -24-[(2-acetamido-2-deoxy-3--3,4,6-tri-0-acetyl-β-D-glu (Copyranosyl)-(1 → 4) -2-acetamido-6-0-acetyl-3-O- (1-carboxy) ethyl- 2-deoxy-β-D-glucopyranosyloxy] -3-methoxycholane (TYC2)   For a solution of TYC21 (unpurified, 0.39 g) in pyridine (10 ml), acetic anhydride (0.86 ml, 9.12 m mol) is added. The reaction mixture is stirred at room temperature for 2 hours. Ethyl acetate (100 ml) And 2M hydrochloric acid (100 ml) are added. The reaction mixture is stirred for 1 hour at room temperature. Organic layer Was separated and washed with water (50 ml), and saturated brine (50 ml), and then dried (Na_Two SO_Four), Concentrated. The crude product (0.41 g, 87%) was used in the next step without further purification Used. TLC (MeOH: DCM 1: 9) Rf 0.47.7. Coupling method to support-binding peptide, elimination method of sugar protecting group, and resin General method for the cleavage method from   R-P, the basic unit of Fmoc-protected resin-binding peptide, is first expanded in DMF. Let moisten (2 mL, 380 rpm, 0.5 hours). Cleave and remove the Fmoc group (2 mL of 20% piper Gin in DMF, 380 rpm, 0.5 hour). Then wash the deprotected R-P 4 times with new DMF Clean (2 mL, 380 rpm, 5 minutes / wash once). Then R-P (1.0 equivalent) is converted to S-L basic unit (1.0-3.0 eq) free acid in the presence of an activating or coupling reagent for about 4 hours In the meantime, process while stirring. One of four different coupling reagents and solvent ("Coupling cocktails") are used. Immediately That is, EEDQ-DCM, HATU-DIPEA / DMF, HBTU-DIPEA / DMF, or PyBOP-DIPEA / DMF You. (See FIG. 3 showing the structural formula of the activating reagent.)   After the coupling reaction has been completed, the coupling mixture is removed by filtration. Wash the resin four times with 2 mL each of fresh DMF (380 rpm, 5 minutes / wash once) . Substituents protecting the sugar (for example, acetyl group, pivaloyl group) are methanol -D Use a solution in which sodium methoxide is mixed in a mixed solvent consisting of MF (1: 1, v / v). By treating the resin for about 15-24 hours with stirring (380 rpm) Removed. After filtration, wash the resin four times with 2 mL fresh methanol-D Wash with MF mixed solvent (380 rpm, 5 minutes / wash once), and then wash 4 times with 2 mL aliquots of DCM (3 80 rpm, 5 minutes / wash once).   The dichloromethane was removed, and the sugar-deprotected resin was removed using a 90% aqueous TFA solution. (2 mL, 380 rpm, 2 hours). The desired product is obtained from the product solution. It is. The remaining resin is 90% TFA-H_TwoThe second process may be performed using O (1 mL, 380 rpm, 15 minutes). In this way a second product solution is obtained, which is initially Combine with the product solution of Trifluoroacetic acid-water solvent at 40 ° C It is removed by evaporation in a block heater.   Addition of diethyl ether (4 mL) precipitates the product lipoglycopeptide You. The resulting mixture is centrifuged (4000 rpm, 10 minutes) at 4 ° C. Then remove the solvent Aspirate with pet and remove, then wash the product with another 2 mL of diethyl ether Cleanse. Repeat the centrifugation step and the pipette suction step once or twice or more. Final The desired product is isolated and dried in a vacuum desiccator.   These general steps are illustrated in FIG. 2 and more particularly illustrated in FIG. 6 or FIG. Have been.   The solid support-bound peptide may be, for example, a Merrifield resin (polystyrene / 1% (Divinylbenzene copolymer) and acid-labile chlorotrityl linker It is formed using a known method. Alternatively, building units can be supplied commercially. For example, from an advanced ChemTeck (Kentucky, U.S.A.) You can do it.   The following describes how to couple the peptide carried by the support to the sugar carried by the lipid. In the following, a specific example of the step of removing the protecting group and the cleavage step will be described. 7.1 Solid phase synthesis of peptidoglycan monomers 52 and 53   The solid phase synthesis of the peptidoglycan monomer analogs 52 and 53 is illustrated in FIG. ing. The sugar 8 carried by the lipid is AKA- (Clt)-which is a chlorotrityl linker. Coupling to the tripeptide bound to the polystyrene resin via (PS). Fat Quality The monosaccharide 51 being transported is a polystyrene resin grafted with polyethylene glycol. As well as coupling to the tripeptide bound to the fat AKA- (PEG)-(PS) , AKA- (Clt)-(PS). The solid phase coupling reaction is glycoca Proceed using 2 equivalents of ribonic acid 8 or 51 and 2 equivalents of coupling reagent. You. The reaction mixture is subsequently shaken for 4 hours at room temperature.   51-AKA- (Clt)-(PS), which is the product of coupling a resin-bound monosaccharide And 51-AKA- (PEG)-(PS) are treated with a 90% aqueous solution of trifluoroacetic acid. To complete the overall deprotection reaction. Monosaccharide pebrigoglycan monomer 52 is obtained by precipitation with diethyl ether [Fab MS: 819 (M-H + 2 Na)⁺].   The disaccharide-coupled product 8-AKA- (Clt)-(PS) is first treated with sodium methoxa The acetic acid protecting group can be removed by treatment with an amide. Followed by 90% T The FA performs a cleavage step and a peptide deprotection step. Finally disaccharide peptide Lican monomer 53 is precipitated using diethyl ether [Fab MS: 991 (M-H + 2 Na)⁺]. Coupling by each combination of solid support and coupling reagent Is determined by RP-HPLC analysis of the isolated product. RP-HPLC analysis Gradient solvent (10% to 50% acetonitrile) on a C18 column (5 micron, 4.6 x 250 mm). UV (205 nm) and ELSD (evaporative light scattering analysis). Monitor). The results (percent yield) are summarized in Table 1. You. 7.2 Solid-phase synthesis of peptidoglycan monomer 54   Another method for synthesizing peptidoglycan monomers is shown in FIG. F The basic unit of the peptide protected by moc, Fmoc- [L-Ala]-[L-Lys]-[D-Ala]-(R ) Is removed in the same manner as described in the general method. Deprotected peptide To pentafluorophenyl (Pfp) ester of lipid-sugar using Dhbt-OH Let me The protecting group elimination step and the cleavage step were performed as described in the general method, and Get. 8. Lipoglycopeptide synthesis assay   The assay for the peptidoglycan polymerization reaction was performed by Mirelman et al. In Biochemist. ry (1976) 15: 1781-1790, and by Allen et al., FEMS Mi crobiol. Lett. (1992) 98: 109-116. Simple Simply put, letting E. coli (ATCC Cat. No. 23226) permeate with ether. Therefore, radioactively labeled radioactively labeled cell wall precursors and exogenously added radioactively labeled Uncleared cell wall precursors can cross the bacterial cell wall. UDP Lamil-pentapeptide (UDP-N-acetylmuramyl-L-Ala-D-Glu-meso-diaminopi Meryl-D-Ala-D-Ala) screens were prepared according to published methods. cereus (ATCC Cat. No. 11778). Kohlrausch & Holtje, F EMS Microbiol. Lett. (1991) 78: 253-258; Kohlrausch et al. Gen. Mcrobiol . (1989) 153: 1499-1506. Bacterial proteins are described in Anal. Biochem. (1 976) 72: 248 and examined by the method of Bradfoed.   Polymerization reaction assay 96-well filter-bottom plate (Millipore GF / C- Cat. No. Perform with MAFC NOB 10). In 100 μl of the final assay solution, Wells are 50 mM Tris-HCl (pH 8.3), 50 mM NH_FourCl, 20 mM MgSO_Four* 7H_TwoO, 10mM ATP (disodium salt), 0.5 mM β-mercaptoethanol, 0.15 mM D-aspara Formic acid, 0.001 mM UDP-N-acetyl [¹⁴C] -D-Glucosamine (DuPont / N.E.N.-307 mCi / mmol), 0.01 mM UDP-MurNAc-pentapeptide, 100 μg / mL tetracycline And 10 μg / well of ether-treated bacterial proteins.   Dissolve the new test compound in distilled water and screen at a final assay concentration of 100 μg / mL. Lean. All raw materials except radiolabel and isolated natural pentapeptide Conversion Chemicals are available from Sigma.   The reaction was performed by leaving a 10 μl aliquot of bacterial protein in assay buffer. Start by adding to the wells containing all the other reagents. Cover the plate And mix for 30 seconds, then incubate at 37 ° C for 2 hours. Ice-cooled 20% TCA ( 100 μl) is added to each well. Gently mix the plate (60 seconds) Cooling (4 ° C) for 30 minutes to ensure precipitation of all peptidoglycan Become fruit.   Place the plate under vacuum filtration conditions on a Millipore manifold and perform filtration. And then wash 4-5 times with 200 μL / well of 10% TCA. Cinchle filter Punch into a container, fill with Cytoscint (ICN), cap and shake. Corn is counted using a Beckman LS6000 spectrometer. Peptidoglyca To¹⁴The percent inhibition of C-label introduction is indicated by the wells of control (total loading). Containing 300 μg / mL vancomycin that can completely inhibit Calculated from wells in the ground (blank). All wells are duplicated And it usually differs by <20%. The concentration response curve for vancomycin is Positive control (IC for inhibition of transduction₅₀= 2.4 ± 0.2 μg / mL, n = 7) Are arranged in a row.   Assay results show that at least three of the libraries show inhibitory activity are doing. Decoding the library shows that all three compounds are the same as shown below. It is clear that they have the same S-L basic unit structure.   The peptide portions of the three compounds showing inhibitory activity are L-S- (L-Ala-D-iso-Gln-L-Ly s-D-Ala-OH, L-S- (L-Lys-D-Ala-D-Ala-OH), and L-S- (Gly-D-iso-Gln-L-Lys-D- Ala-D-Ala-OH).   It is certain that other embodiments of the invention will be apparent to those skilled in the art. like this Embodiments are not specifically described in this specification, but may not be described. Nevertheless, they are not limited by the particular embodiments described herein, but The scope and spirit of the present invention is limited only by the appended claims.

────────────────────────────────────────────────── ─── Continuation of front page    (81) Designated countries EP (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE, IT, L U, MC, NL, PT, SE), OA (BF, BJ, CF) , CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (GH, KE, LS, MW, S D, SZ, UG), UA (AM, AZ, BY, KG, KZ , MD, RU, TJ, TM), AL, AM, AT, AU , AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, G B, GE, GH, HU, IL, IS, JP, KE, KG , KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, N O, NZ, PL, PT, RO, RU, SD, SE, SG , SI, SK, TJ, TM, TR, TT, UA, UG, UZ, VN, YU (72) Inventor Chen Anna             United States of America New Jersey La             -Way Madison Avenue 852 (72) Inventor Liu Dashan             United States of America New Jersey             Ost Brunswick Rice Run               70 (72) Inventor Zhang Ting Yau             United States of America New Jersey             Dison Barlow Road 26

Claims (1)

Claims: 1. A solid-phase lipoglycopeptide library comprising a plurality of unique substances represented by the following chemical formula. In this formula, the substituent R is a solid support, the substituent P is one or more amino acids, peptides, or polypeptides, the substituent S is one or more sugars, and the substituent L is one. Or more lipids. 2. The library of claim 1, wherein there are more than one copy of each unique substance. 3. The library according to claim 1, wherein at least one substituent of each substituent in the chemical formula is covalently bonded to at least one substituent of an adjacent substituent. 4. The library according to claim 3, wherein the same sugar having the substituent S is covalently bonded to at least one substituent of the substituent P and at least one substituent of the substituent L. 5. The library according to claim 3, wherein at least one sugar of the substituent S is covalently bonded to the lipid of the substituent L via a covalent bond containing a glycosidic bond. 6. The library according to claim 3, wherein at least one sugar having the substituent S is covalently bonded to the lipid having the substituent L via an acetamido bond of the alpha-anomeric hydroxyl group. 7. The method according to claim 4, wherein at least one sugar of the substituent S is covalently bonded to the substituent of the substituent P via an alpha-acetamide portion or an alpha-propionamide portion of the C-3 hydroxyl group. Library. 8. The library according to claim 3, wherein the covalent bond is formed via a linker. 9. The library according to claim 4, wherein the linker is unstable. 10. The library according to claim 4, wherein the linker is acid-labile or photolabile. 11. The library according to claim 4, wherein at least one of the substituents P is bonded to the resin via a halotrityl moiety. 12. The library according to claim 11, wherein the halotrityl moiety is a chlorotrityl moiety. 13. The library of claim 4, wherein at least one of the substituents P is attached to the resin by an alpha-bromo-alpha-methylphenacyl moiety. 14. The library of claim 3, wherein the covalent linkage comprises one or more amine, ether, thioether, ester, thioester, amide, acetamido, phosphate, phosphonate, phosphinate, or sulfate linkage. 15. The library of claim 14, wherein the covalent linkage comprises one or more glycosidic linkages. 16. The library of claim 1, wherein the solid support comprises an insoluble polymer, metal or glass surface. 17. The library according to claim 1, wherein the amino acids, peptides, or polypeptides are all hydrophilic amino acid residues or the hydrophilic amino acid residues are dominant. 18. The library according to claim 1, wherein the amino acids, peptides, or polypeptides are all hydrophobic amino acid residues or the hydrophobic amino acid residues are dominant. 19. The library according to claim 1, wherein the saccharide is a monosaccharide, a disaccharide, or a polysaccharide. 20. The library of claim 19, wherein the monosaccharide comprises hexose, pentose, their deoxy analogs, their dideoxy analogs, their azide-substituted analogs, or their amino-substituted analogs. 21. The library of claim 19, wherein the disaccharide or polysaccharide comprises hexoses, pentoses, their deoxy analogs, their dideoxy analogs, their azide-substituted analogs, their amino-substituted analogs, or combinations thereof. 22. The library of claim 1, wherein the lipid is a saturated, unsaturated, or polyunsaturated, linear, branched, or cyclic aliphatic substituent containing 2-60 carbon atoms. 23. The following chemical formula A method for preparing a solid-phase lipoglycopeptide library provided with a plurality of unique substances represented by the following, (a) one or more substituents R, P and FG represented by the following chemical formula Step of providing _one . In this formula, R is a solid support that binds a substituent P containing one or more amino acids, peptides, or polypeptides, and at least one of the substituents P is capable of participating in a bond-forming reaction. and a first functional group FG _1. (b) providing one or more substituents S, L and FG _{2 represented} by the following formula: In this formula, S is one or more sugars, and at least one of the sugars is a sugar that binds a substituent L containing one or more lipids, and the substituent S at least one sugar is provided with a second functional group FG ₂ capable of participating in bond formation reactions. (c) so as to proceed with one of the substituents P having a first functional group FG _1, the bond forming reactions to form a bond between the sugar substituents S having a second functional group FG _2, one or more substituents R, P and FG ₁ and one or more substituents S, the step of coupling the L and FG _2. A method that includes 24. The method according to claim 23, further comprising a step of immersing the solid support in an organic solvent before the bonding step. 25. Prior to bonding step, the method according to the range 23 claims further comprising the step of removing the protecting group in the first functional group FG _1. 26. The method of claim 23, wherein the coupling step comprises providing an activating reagent to aid in the progress of the bond formation reaction. 27. The method according to claim 23, comprising removing the protecting group on the substituent P, S, or L. 28. The method of claim 23, further comprising screening the resulting lipoglycopeptide library for one or more active agents. 29. The method according to claim 23, further comprising the step of cleaving the substituent P from the substituent R. 30. The method according to claim 29, further comprising the step of recovering one or more substances remote from the substituent R. 31. The following chemical formula: A method for preparing a solid-phase lipoglycopeptide library provided with a plurality of unique substances represented by the following, (a) one or more substituents R, P and FG represented by the following chemical formula Step of providing _one . In this formula, R includes a solid support that binds a substituent P containing one or more amino acids, peptides, or polypeptides, and at least one of the substituents P participates in an initial bond formation reaction. and a first functional group FG ₁ as possible. (b) one represented by the following formula or more substituents S, providing a FG ₂ and FG _3. Note In this formula, S is one or more of a sugar, have a second functional group FG ₂ at least one sugar can be involved in the initial binding formation reaction of the substituent S, also and a third functional group FG ₃ capable of participating in bond formation reaction ensues same sugar or a different sugar of the substituent S. and (c) coupling allowed to proceed first bond formation reaction to form between the sugar part and the substituent S having a second functional group FG ₂ substituents P having a first functional group FG _1, to provide one or more of the initial bond forming reaction products, one or more substituents R, P and FG ₁ and one or more substituents S, and FG ₂ and FG ₃ Combining. (c) providing one or more substituents L and FG _{4 represented} by the following formula: Note In this formula, substituent L is a one or more lipids, a fourth functional group F G ₄ capable of participating in at least one lipid ensues bond formation reactions of the substituents L I have. (d) so as to advance the subsequent forming bonds bond forming reactions between one and lipid substituent L with a fourth functional group FG ₄ substituents S having a third functional group FG _3, one or more of the initial bond forming reaction product with one or more substituents L and FG ₄ and bonding the. A method that includes 32. One or more of the substituents S, L and FG ₂ is one or more substituents R, and P and FG _1, claims to bind at room temperature in a solvent in the presence of activating reagents 31. The method according to 31. 33. The method according to claim 31, wherein FG ₂ comprises a pentafluorophenyl ester. 34. The method according to claim 31, wherein the activating reagent comprises EEDQ, HATU, HBTU, or PyBOP. 35. The method of claim 31, wherein FG ₂ comprises a carboxylic acid and FG ₁ comprises an amine. 36. The method according to claim 31, wherein FG ₁ is located on one side chain of the substituent P. 37. The following chemical formula The following chemical formula obtained by cleaving the substituent P from the substituent R from the substance represented by A composition represented by the formula: In this formula, the substituent R is a solid support, the substituent P is one or more amino acids, peptides, or polypeptides, the substituent S is one or more sugars, and the substituent L is one or more. Contains more lipids. 38. A synthetic composition represented by the following chemical formula: In this formula, the substituent P includes one or more amino acids, peptides, or polypeptides, the substituent S includes one or more sugars, and the substituent L includes one or more lipids. 39. The composition of claim 38, wherein P comprises two or more amino acids in the sequence. 40. The composition of claim 39, wherein P comprises one or more peptides or polypeptides. 41. The composition of claim 40, wherein each peptide or polypeptide comprises three or more amino acids in the sequence. 42. The composition of claim 38, wherein the lipid comprises 2-60 carbon atoms and is a saturated, unsaturated, or polyunsaturated linear, branched, or cyclic aliphatic substituent. 43. The composition of claim 38, wherein the lipid comprises 4-50 carbon atoms and is a saturated, unsaturated, or polyunsaturated linear, branched, or cyclic aliphatic substituent. 44. The composition of claim 38, wherein the lipid comprises 10-30 carbon atoms and is a saturated, unsaturated, or polyunsaturated linear, branched, or cyclic aliphatic substituent. 45. The composition according to claim 38, wherein the lipid comprises a substituted or unsubstituted aromatic or heteroaromatic substituent. 46. The composition according to claim 38, wherein the composition is synthesized by a combination method. 47. The composition of claim 38, wherein the composition does not match the structure of the naturally occurring lipid-bound glycopeptide intermediate. 48. The composition according to claim 38, wherein when the substituent L comprises one lipid, it does not comprise an andecaprenyl group. 49. The composition of claim 38, wherein at least one sugar of substituent S is covalently linked to at least one lipid of substituent L. 50. the covalent bond is a glycoside ether bond, phosphate, pyrophosphate, phosphinyl, phosphanyl, phosphonate, phosphonyl, phosphono, phosphino, phosphanoacetate, phosphonyl formate, phosphoramidyl phosphorothionate, phosphonyl sulfonate, or 49. The composition according to claim 48, comprising a phosphonyl sulfonate linkage. 51. A compound represented by the following chemical formula: Wherein L ¹ contains a lipid substituent, P ¹ contains one or more amino acids, peptides or polypeptides, and G ¹ , G ² , G ³ , G ⁴ , or G ⁵ are each independently A substituted or unsubstituted, branched or unbranched alkyl group, alkoxy group, alkenyl group, C1-C8 acyl group, acetyl group, alkoxycarbonyl group, hydroxyalkyl group, carboxyalkyl group, or substituted Or an unsubstituted aromatic or heteroaromatic substituent, or hydrogen, and X ¹ , X ² , X ⁵ , X ⁶ or X ⁷ are each independently oxyalkyl, amine, ether, Thioether, ester, thioester, amide, acyl, acetamido, phosphate, phosphinate, pyrophosphate, sulfate, azide, hydroxyl or functional group containing hydrogen May be, except its functional groups azido, hydroxyl or is absent G substituents attached For hydrogen,, X ³ or X ⁴ are each independently, an amine, ether, thioether, ester, thioester , Amide, acetamido, acyl, phosphate, phosphinate, pyrophosphate, a functional group containing a sulfate substituent, y is 0, 1, 2, or 3, m, n, v, w, or z May be independently 0, 1, 2, or 3, provided that the sum of m and n is not greater than 3, and the sum of v, w, and z is not greater than 5, and q or u is Independently, it can be 1, 2, or 3, provided that the sum of q and u is not greater than 5, provided that the compound is 2-N-acetyl-1-α-O-allyl-4, 6-O-isopropylidenemuramyl-L-alanyl-D-glutamine benzyl ester, (2R)- Benzyl 2- [N- (2′-N-acetyl-1′-α-O-allyl-4 ′, 6′-O-acetylmuramyl-L-alanyl) amino] -4-cyanobutanoate, ( 2R) benzyl 2-N-acetyl-1'-α-O-allyl-4,6-O-isopropylidenemuramyl-L-alanyl) amino] -4-cyanobutanoate, (2R, 2 "R) -Benzyl 2- [N- [2'-N-acetyl-1'-α-O-[(N, N-diisopropylamino) (butoxy) phosphinyl] -4 ', 6'-di-O-acetylmuramyl -L-alanyl] amino] -4-cyanobutanoate, (2R, 2 "R) -benzyl 2- [N- [2'-N-acetyl-1'-α-O-[[2" -carbo Benzoxy-2 "-(pentyloxy) ethoxy] (benzyloxy) phosphoryl] -4 ', 6'-di-O-acetylmuramyl-L-alanyl] amino] -4-cyanobutanoate, (2R, 2 "R) -2- [N- [2'-N-acetyl-1'-α-O-[[2" -carboxy-2 "-(pentyloxy) ethoxy] hydroxyphosphoryl] muramyl-L-alanyl Amino] -4-cyanobutanoate or pe It is not a natural substrate for either the transglycosylase activity of the nicillin binding protein or the N-acetylglucosaminyltransferase activity of the murG gene product. 52.X ¹ G ¹ or X ² G ² may be located at position 2 or 5, X ³ P ¹ is located at position 3, and X ⁴ L ¹ is located at position ¹ of the ring 52. The compound of claim 51, wherein: 53.X ¹ G ¹ , X ⁵ G ³ , X ⁶ G ⁵ , or X ⁷ G ⁴ is a hydroxyl group, HOCH ₃ CH _2- , acetamide, phthalimide, benzoyl, alkoxybenzoyl, alkoxycarbonylalkyl, carboxyalkyl, or pivalyl group 52. The compound according to claim 51, which may be: 54.P ¹ is one or composition according to claim 51 of claim containing more peptides or polypeptides. 55. The composition of claim 54, wherein each peptide or polypeptide comprises three or more amino acids in the sequence. 56.The composition of claim 51, wherein L ¹ comprises 4-30 carbon atoms and comprises a saturated, unsaturated, or polyunsaturated linear, branched, or cyclic aliphatic substituent. . 57.L ¹ A composition according to claim 51 claims including that substituted or unsubstituted aromatic or heteroaromatic substituent. Covalent bond 58.X ⁴ is, glycoside ether bond, phosphates, pyrophosphates, phosphinyl, phosphanyl, phosphonate, phosphonyl, phosphono, phosphino, phosphonium Fano acetate, phosphonyl formate, phosphoramidate Gilles phosphorothioates, phosphonyl 52. The composition of claim 51 comprising a sulfonate or phosphonyl sulfonate linkage.