DE19709787A1 - Oligosaccaride and their derivatives as well as a chemo-enzymatic process for their production - Google Patents

Abstract

The invention relates to novel oligosaccharides and the derivatives thereof in addition to a general method for stereo divergent production of oligosaccharides from easily accessible simple glycosides, wherein a further saccharide element is stereo selectably created from the aglycone constituent thereof by means of chain elongation reactions. This is achieved by (optional) chemical addition of an aldehyde equivalent to a C=X-double bond in the aglycone, followed by diastereo-selective enzymatic addition of a nucleophile aldol donor to the glycosylated aldehyde in the presence of various stereo-specific aldolases. The resulting oligosaccharides, which carry an additional ketose unit on the reducing end when DHAP-dependent aldolases are used, and their corresponding phosphate esters and suitable derivatives thereof are useful as constituents of precursors for pharmaceutically active substances.

Description

The invention relates to new oligosaccharides and their derivatives and a general Process for the stereodivergent production of oligosaccharides from easily accessible Lichen simple glycosides, from their Aglykonteil by chain extension stereoselectively, another saccharide unit is built. This will achieved by (optional) chemical addition of an aldehyde equivalent to an C = X double bond in the aglycone, followed by a diastereoselective enzymati add a nucleophilic aldol donor to the glycosylated aldehyde in Presence of various stereospecific aldolases. The resulting oligosac charide, which causes additional ketosis when using DHAP-dependent aldolases Wear unit at the reducing end, as well as their corresponding phosphate esters and suitable derivatives are of interest as components or pharmaceutical precursors active ingredients.

Oligosaccharides play an eminently important role as information carriers of high Density of information in many biological recognition processes such as B. cells growth, differentiation, aggregation and migration. With countless people diseases, oligosaccharide-receptor interactions are of fundamental importance gender meaning such as B. in immunological defense reactions, tumor crane or infections caused by parasites, bacteria or viruses. By Oligosac Charid-derived structures are therefore increasingly of interest as potential phar pharmaceuticals, biological communication channels - especially in the case of illness such as acute or chronic inflammation, asthma, immunological er diseases, tumor metastasis, graft rejection, infectious diseases and cardiovascular diseases - to moderate or block.

Only in exceptional cases can oligosaccharides be obtained from natural sources in pure form and in large quantities, which is why in most cases you have to rely on targeted syntheses. Although carbohydrate chemistry is relatively highly developed, there is no universal solution for the synthesis of oligosaccharides (cf. RR Schmidt in Comprehensive Organic Synthesis, ed. BM Trost and I. Fleming, Pergamon Press, New York 1991, Vol. 6, p. 33-64; O. Lockhoff in Methods of Organic Chemistry (Houben-Weyl), ed. H. Hagemann and D. Klamann, Thieme-Verlag, Stuttgart 1992, Vol. E14a / 3, pp. 621-1077; K. Toshima , K. Tatsuda, Chem. Rev. 1993, 93, 1503-1531). The tremendous variety of linkage options between the multifunctional monosaccharides makes the stereoselective synthesis of oligosaccharides extremely complex for both chemical and enzymatic synthesis processes and requires a number of problems to be overcome:

• a) each sugar module and each type of link between similar or different building blocks represents an individual synthesis problem;
• b) the building blocks allow a variation according to type (sequence) and size (Pentoses / hexoses) and contain one
• c) high density of chemically identical functionalities with very similar ones Reactivity;
• d) the synthesis possibilities are due to the different availability of the Monosaccharide components (mostly from nature) drastically limited;
• e) many naturally occurring monosaccharide building blocks contain special ones Additional or different functionalities (e.g. ketoses, sugar acids, Aminosugar, esters of inorganic acids);
• f) the variation of the linking location between building blocks (regioselectivity) requires the targeted provision of a free one for chemical processes OH group on the acceptor and protection of all other OH groups, whereby
• g) the protective groups in the acceptor have a controlling influence on the reactivity exercise the acceptor OH group;
• h) the variation of the linkage at the anomeric carbon atom (diastereoselectivity with regard to the a / b configuration) must by the decisive influence the protective groups in the donor are controlled stereochemically, being it
• i) additional kinetic and thermodynamic influences on the diastereo selectivity (anomeric effect) to be taken into account by the conduct of the reaction applies what  
• k) by the type of activation of the anomeric center in the glycosyl donor by introducing a suitable escape group and choosing a catalytic that promoter can be influenced;
• l) Especially with longer synthesis sequences, the targeted selection requires of protective groups of different reactivity (permanent and tempo rare), sophisticated protection group strategies and complex manipulations;
• m) Yields in the formation of the glycosidic bond are often due to the possible side reactions relatively small, which u. a. also
• n) in the sensitivity of the glycosidic (acetal) binding to acid-catalyzed hydrolysis is justified;
• o) enzymatic processes based on biosynthesis require the agile activation as unstable nucleotide phosphate sugar and
• p) access to a range of donor / acceptor specific and as kapri ziös known enzymes for glycosyl transfer, of which
• q) only a few are commercially available and expensive;
• r) Transglycosylations with cheaper glycosidases never notoriously deliver yields and
• s) are unattractive due to the high separation effort involved in product isolation.

Oligosaccharides built up from the naturally occurring monosaccharides also because of their high biological metabolism rate for therapeutic use unsuitable for set purposes.

The invention describes a widely applicable method for the rapid, stereoselective generation of a variety of complex oligosaccharides and of metabolically stable non-natural derivatives from cheap or easily accessible starting materials. It has surprisingly been found that glycosylated aldehydes, which are conveniently and efficiently accessible due to their simple structure using conventional synthetic methodology, can be stereoselectively converted to oligosaccharides enlarged by one saccharide unit by means of certain chemical and / or enzymatic processes for chain extension by means of CC linkage, where elaborate protection group techniques can be largely avoided. Since several chirality centers are newly created, this procedure allows the controlled stereodivergent synthesis of a large number of diastereoisomers (2 ⁿ for n new centers) from the same aldehyde starting material.

The invention relates to compounds of the general formula (I)

as well as pharmaceutically acceptable salts thereof,
in which
A represents CH ₂ or a radical of the formula

stands,
B represents CH ₂ , a bond or a radical of the formula

stands,
where A and B should not simultaneously represent a CH ₂ group,
and
R represents hydrogen or phosphate,
in which
R ^{1 represents} an optionally substituted glycosyl radical,
R ² and R ^{3 may be the} same or different and represent hydrogen, hydroxy, hydroxyalkyl, glycosyloxy, glycosyloxyalkyl, alkoxyalkyl, dialkoxyalkyl or azido or
R ² and R ³ together with the carbon atom to which they are attached represent a 5- or 6-membered saturated ring which can contain up to 2 oxygen atoms, where the ring can be substituted up to twice by lower alkyl, and
R ^{4 represents} hydrogen, alkyl, alkyloxycarbonyl, carboxy or glycosyloxyalkyl,
X represents -O-, -OCH ₂ -, -N (alkanoyl) -, -S-, -SCH ₂ - or a bond,
R ^{1 ',} independently of R ¹ , can have the meanings given for R ¹ ,
R ^{3 '} independently of R ³ can have the meanings given for R ³ and
X 'can have the meanings given for X independently of X.

The compounds of formula (I) contain several stereocenters.  

They can therefore exist in stereoisomeric forms that are either like image and Mirror image (enantiomers), or which are not like image and mirror image (Diastereo mere) behave. The invention relates to both the enantiomers and the di astereomers or their respective mixtures. Mixtures of the enantiomers or the diastereomers can also be known, for example by crystallization sations- or chromatography in the stereoisomerically uniform constituents separate.

Compounds of the formula (I) are preferred
in which
A, B, R, R ¹ and R ^{1 'have} the meaning given above and
R ² and R ^{3 may be the} same or different and stand for hydrogen, hydroxy, hydroxymethyl, glycosyloxy, glycosyloxymethyl, alkoxyalkyl, dialkoxy-alkyl or azido or together represent -OC (CH ₃ ) ₂ OCH ₂ - and
R ^{4 represents} hydrogen, methyl, methoxycarbonyl, carboxy or glycosyloxymethyl,
X represents -O-, -OCH ₂ -, -N (acetyl) -, -S-, -SCH ₂ - or a bond,
R ^{3 '} independently of R ³ can have the meanings given for R ³ and
X 'can have the meanings given for X independently of X.

Examples of glycosyl residues and for the glycosyl residues in the "glycosyloxy" residues and "glycosyloxyalkyl" are erythrofuranosyl, threofuranosyl, ribofuranosyl, Arabinofuranosyl, xylofuranosyl, allofuranosyl, glucofuranosyl, mannofuranosyl, Idofuranosyl, galactofuranosyl, talofuranosyl, erythropentulofuranosyl, fructo furanosyl, altroheptulofuranosyl, glyceromannooctulofuranosyl, erythrogluco  nonulofuranosyl, (methyl-5-deoxy-ribofuranoside) -5-yl, (methyl-6-deoxy-gluco furanoside) -6-yl, (methyl-3-deoxy-glucofuranoside) -3-yl, (methyl-3-deoxy-galacto furanoside) -3-yl, (benzyl-3-deoxy-galactofuranoside) -3-yl, ribopyranosyl, arabino pyranosyl, xylopyranosyl, allopyranosyl, glucopyranosyl, mannopyranosyl, ido pyranosyl, galactopyranosyl, talopyranosyl, fucopyranosyl, rhamnopyranosyl, Fructopyranosyl, Altro-heptulo-pyranosyl, Glycero-manno-octulo-pyranosyl or Erythro-gluco-nonulopyranosyl, (methyl-6-deoxy-glucopyranoside) -6-yl, (methyl-3- deoxy-glucopyranoside) -3-yl or methyl-4-deoxy-glucopyranoside) -4-yl.

One or more hydroxyl groups in the glycosyl residues can be exchanged against other substituents, for example against hydrogen, lower alkyl, Halogen, lower alkoxy, oxa-lower alkyloxy, lower alkenyloxy, cycloalkoxy, Tetrahydropyranyloxy, arylalkyloxy, heteroarylalkyloxy, lower alkyloxy arylalkyloxy, alkylidendioxy, benzylidendioxy, lower alkanoyloxy or Niederalkanoyloxy, which is substituted by halogen, arylcarbonyloxy or Arylcarbonyloxy which is substituted by halogen or lower alkoxy, Lower alkylsulfonyloxy, arylsulfonyloxy, lower alkylarylsulfonyloxy, oxo, amino, Mono- or di-lower alkylamino, formylamino, lower alkanoylamino or Lower alkanoylamino which is substituted by halogen, lower alkynylamino, Arylalkylamino, azido, phthalimido, nitryloxy, mercapto, lower alkylthio, Lower alkanoylthio, carboxy lower alkyloxy, lower alkoxycarbonylalkyloxy, carboxy, Lower alkyloxycarbonyl, lower alkenyloxycarbonyl, benzyloxycarbonyl, Phosphonooxy, dialkyloxy-phosphoryloxy, dibenzyloxyphosphoryloxy, sulfooxy, Alkyloxysulfonyloxy, benzyloxysulfonyloxy or sulfamoyloxy, optionally substituted glycosyloxy or optionally substituted di- or Oligosaccharidyloxy.

Examples of glycosyl radicals in which one or more or all of the hydroxy groups have been exchanged or substituted are 2-amino-2-deoxy-gluco pyranosyl, 2-amino-2-deoxy-galactopyranosyl, 2-acetylamino-2-deoxy-gluco pyranosyl, 2-acetylamino-2-deoxy-galactopyranosyl, 2-azido-2-deoxy-glucopyra nosyl, 2-deoxy-glucopyranosyl, 4-deoxy-glucopyranosyl, 6-chloro-6-deoxy-gluco  furanosyl, 6-deoxy-6-fluoro-glucopyranosyl, 4-chloro-4-deoxy-glucopyranosyl, Glucofuranos-3-ulosyl, 6-thio-glucopyranosyl, 3-thio-galactofuranosyl, 5-acetyl amino-3,5-dideoxy-glycero-galacto-2-nonulopyranosyl-one acid, methyl- (5-acetyl amino-3,5-dideoxy-glycero-galacto-2-nonulopyranosyl) -onate, glucuronopyranosyl, Galacturonofuranosyl, cellobiosyl, maltotriosyl, 3-O- (carboxymethyl) fucopyranosyl, 6-O-phosphoyl-galactopyranosyl, 2,3,4,6-tetra-O-methyl-glucopyranosyl, 2,3,4,6- Tetra-O-acetyl-glucopyranosyl, 2,3,4,6-tetra-O-benzyl-glucopyranosyl, 2,3,4,6- Tetra-O-benzyl-galactopyranosyl, 2,3,4,6-tetra-O-benzyl-mannopyranosyl, 2,3,4-tri- O-benzyl-rhamnopyranosyl, 2,3,4-tri-O-benzyl-fucopyranosyl, 2,3-di-O-allyl-4,6-di- O-benzyl-mannopyranosyl, 2,3-di-O-acetyl-4,6-O-benzylidene-glucopyranosyl, 2,3,5,6-di-O-isopropylidene-mannofuranosyl, 2,3-O-isopropylidene-ribofuranoside, Me thyl- (5-acetylamino-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-glycero-galacto-2-nonulo pyranosyl) -onate or 2,3,6,2 ', 3', 4 ', 6'-hepta-O-acetyl-maltosyl.

"Alkyl" as a separate substituent or as part of other radicals such as hydroxyalkyl, glyco syloxyalkyl, alkoxyalkyl, dialkoxyalkyl, alkoxy, alkanoyl means a straight chain term or branched alkyl radical with up to 8, preferably up to 6 and particularly be preferably up to 4 carbon atoms.

"Lower alkyl" as a separate substituent or as part of other radicals such as Lower alkyloxy means a straight-chain or branched alkyl radical with up to 7, preferably up to 5 and particularly preferably up to 3 carbon atoms.

"Niederalkenyl" as a separate substituent or as part of other radicals such as Lower alkenyloxy represents a straight-chain or branched alkene radical of up to 7, preferably up to 5 and particularly preferably up to 3 carbon atoms.

"Alkylidene" as a separate radical or as part of other radicals such as alkyli dendioxy means a straight-chain or branched alkylidene chain with up to 8, be preferably up to 6, particularly preferably up to 4 carbon atoms.  

"Aryl" as a separate substituent or as part of other radicals such as Arylalkyloxy represents an aryl radical with up to 10, preferably up to 6 carbon atoms, for example for phenyl.

"Alkynyl" as a separate substituent or as part of other radicals such as Lower alkynylamino represents a straight-chain or branched alkynyl radical with bis up to 8, preferably up to 6, particularly preferably up to 4 carbon atoms.

Furthermore, the present invention relates to a process for the production of en antiomerically pure and diastereomerically pure oligosaccharides and their derivatives from glycosides with a non-saccharidic aglycone content by chain-building reactions, characterized in that

• - That one successively coals to a C = X double bond in the aglycone nucleophiles in the presence of a metal and dihydroxyacetone phosphate in Presence of an enzyme added.
• - That one uses allyl glycosides of natural mono- and oligosaccharides.
• - that substituted, deoxygenated, alkylated, acylated, acetalized (etc.) Sugar, aminosugar or sugar acids are used.
• - That one uses O-glycosides.
• - That one uses C-, N- or S-glycosides.
• - That one is substituted as carbon nucleophiles for the first addition or unsubstituted allyl or propargyl halides are used.
• - That one as metals with medium reducing power, preferably zinc, tin or indium.
• - That one carries out the reaction in an aqueous or alcoholic medium.
• - That you choose a reaction temperature of -20 ° to + 80 ° C.
• - That one as carbon nucleophiles for the second addition Dihydroxyacetonphos phat and a fructose-1,6-bisphosphataldolase ([EC 4.1.2.13]), Tagatose-1,6- bisphosphataldolase ([EC 4.1.2]), fuculose-1-phosphataldolase ([EC 4.1.2.17]) or rhamnulose-1-phosphataldolase ([EC 4.1.2.19]).
• - That one uses 4-hydroxy-3-oxobutylphosphonic acid instead of DHAP.  
• - That you have other enzymes for C-C linkage with substrate specificity for Brenz grape acid or phosphoenol pyruvate used.
• - That one carries out the reaction in an aqueous medium.
• - That you add up to 50% organic cosolvent.
• - That one carries out the reaction at pH 6.0 to 8.0.
• - That you choose a reaction temperature of -5 ° to + 40 ° C.
• - That one enzymatically catalyzes the phosphate group from the sugar phosphate esters removed by acidic or alkaline phosphatase.
• - That one as a stabilizing additives heavy metal salts, alkali metal salts or Uses thiol reagents.
• - That one uses the enzymes in immobilized form.
• - That the reaction sequence with a saccharide fixed to a solid phase carries out.

The compounds of the formula (I) can, as a rule, consist of three continuous key steps. A schematic overview in Scheme 1 may clarify this without being restrictive.  

Chemoenzymatic production of oligosaccharides

Chemoenzymatic production of oligosaccharides

The process according to the invention for the production of enantiomerically pure and diastereomerically pure oligosaccharides and their derivatives, in particular of compounds of the general formula (I), is characterized by the following three key steps:

• (1) First, in the first key step according to standard methods, a glycoside that contains a polar addition-capable double bond in the aglycon part, typically an aldehyde, ketone or ester-carbonyl grouping or a synthesis equivalent familiar to the person skilled in the art, from which this function is used before the next reaction step can be easily generated.
  Such a suitable glycoside can e.g. B. can be obtained by preparing compounds of the formula (IV) by known processes,
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given above and
  R ⁵ and R ^{6 can be the} same or different and represent hydrogen, alkyl or aryl,
  and converting them into compounds of the formula (V) by cleavage of the double bond, for example by the action of ozone,
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given above.
• (2) In the second key step, a C-nucleophilic reagent with CC linkage is added to this double bond, a heteronucleophile, which is typically an alcohol, being produced from the original double bond. The reagent itself must contain an aldehyde function in masked form, such as, for example, an olefin, acetal, an alcohol, ester, a carboxylic acid or other structures equivalent to those known to those skilled in the art.
  Reacting compounds of the formula (V) with organometallic reagents which have a double bond gives, for example, compounds of the formula (VI)
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given above and in the
  Y represents -CH ₂ -, CH (CH ₃ ) -, -C (CH ₃ ) ₂ - or a bond and
  Z for = CH ₂ or = C = CH ₂
  stands.
• (3) In the third key step, the aldehyde of the formula (VII)
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given above,
  released by a corresponding chemical reaction from the masked aldehyde function of the compounds obtained in the second key step, and the aldehyde obtained is then enzyme-catalyzed in the sense of an aldol reaction by means of a specific aldolase dihydroxyacetone phosphate (DHAP) with CC linkage, with cyclization via the previously generated Heteronucleophile to the ketone group of the DHAP unit forms a typical sugar ring structure of the formula (I).

In a modification of this procedure, a heteronucleophile can optionally also be contained directly in the aglycon part, whereupon the second step can be omitted. On the other hand, step (2) can also be repeated several times in the sense of the invention will.

For the glycosides to be produced in the first step, the well-developed glycosidation methods according to Fischer or the numerous variants of the König-Knorr synthesis can be used (see the overviews on the state of the art above). In this step, the glycosidic portion can be selected according to its constitution and the glycosidic linkage. In addition to the natural aldoses and ketoses (typically with four to 8 carbon atoms in the chain), those with a non-natural configuration or constitution (for example aza, thia, phosphate or carbazucker) and differently substituted ones (for example O- or C-alkylated, O-acylated, deoxy- or amino sugar) or oxidized (glyconic and glycuronic acids, ketoaldoses, dial cans, etc.) descendants can be used. The aglycon portion can also be varied widely, but must contain a polar double bond (C = O, C = N, etc.) which can be added, which is typically an aldehyde, ketone or ester carbonyl grouping. This can advantageously also be represented by a synthesis equivalent familiar to the person skilled in the art, from which this function can easily be generated before the next reaction section. The aglyconic part can be varied in its constitution (linear, branched, substituted), size (typically C ₂ -C ₄ ), glycosidic linking position (typically alpha or beta to the aldehyde) and possibly its absolute configuration.

For example, olefins are very good and widely commercially available synthesis equi valente, which is best used with the methods to tie the Glycoside bond and release of a C = O double bond are compatible. As The simplest and typical case is the allyl glycosides, of which some are also commercially available. Alternatively, other aldehydes can also be used precursors are used such as glycosylated acetals, alcohols, esters, carbon acids and other synthetic equivalents familiar to the person skilled in the art.

In addition to the typical natural O-glycosidic linkage of the aglycon other links are also used for the method according to the invention such as, for example, N-, S- or C-glycosides, their preparation to the person skilled in the art is common and which is a significantly higher metabolic compared to the O-glycosides Have stability.  

For the purposes of the invention, the 1-O- and 2-O- Glycosides of 3-butene-1,2-diol and compounds to be derived therefrom, since here a times the linkage point in the glycosyl acceptor can be optionally determined and for second, the heteronucleophile is introduced for later ring formation. Butene diol is commercially available in both racemic and enantiomerically pure form bar.

Examples of the range of variation of the connection types in question are in Scheme 2 listed.

The metal-mediated allyl transfer from allyl halides (reaction of the Barbier type; for SdT, see CJ Li, Chem. Rev. 1993, 93, 2023-2035; TH Chan et al. Can J Chem. 1994, 72, 1181-1192; Lubineau et al. Synthesis 1994, 741-760) as an excellent method for protecting group-free CC linkage, in particular zinc, tin or indium metal because of the high reaction selectivity as well as aqueous and / or alcoholic solvent systems are preferred because of the good solubility of the sugar derivatives. This is also surprising for the person skilled in the art insofar as, despite the synthesis conditions essential for this type of reaction (usually achieved by adding mineral acids such as HCl or acidic salts such as NH ₄ Cl) and the reaction-related formation of Lewis acidic by-products (two or tetravalent metal halides) the acid-labile glycoside bond is completely preserved. The allyl halides used can be variably substituted on the double bond.

Also surprising for the person skilled in the art, when using propargyl bromides in combination with tin or indium metal in aqueous and / or alcoholic solvent systems, good selectivity ( ³ 50%), the allenyl adducts are formed in addition to the corresponding propargyl compounds. In the case of ozonolytic cleavage, the former react in the sense of vinyl derivatives to aldehydes glycosylated in the a-position.

Alternatively, the same compounds can also be added well, for example of allyl or vinyl Grignard reagents to protected glycosyl aldehydes (chemo selectively even with O-acetyl protective groups).

The reaction of the second process section generates a at the addition point new chirality center. Both of the above methods provide Diastereomers at comparable proportions. Separation of the diastereomers on this or the next stage can be avoided if asymmetric cal allyl silicon, allyl tin or allyl boron reagents is used. This However, processes can only be used in aprotic organic solvents and therefore require the introduction of suitable protective groups in the glycoside.

Examples of the range of variation of the connection types in question are in Scheme 3 listed.

The third step of the method according to the invention uses an enzymatic one Aldol addition to complete the transformation of the aglycone into one Sugar residue. For this purpose, the aldehyde function from the one in the preceding is first Step introduced synthetic equivalent generated and enzyme-catalyzed an aldol Donor added as a nucleophile. The structure of the aldol donor should be chosen so that it matches the substrate selectivity of the aldolase. According to the invention Processes are highly diastereoselective and allow synthesis of all diastereoisomeric oligosaccharides to be derived from this step as Glycosylketoses and their derivatives in high chemical and optical purity.

Four DHAP-dependent aldolases with different ones can preferably be used for this purpose Diastereospecificity can be used optionally, namely the D-fructose-1,6-diphos phataldolase ([EC 4.1.2.13]; D-threo- = (3S, 4R) configuration), D-Tagatose-1,6-di phosphate aldolase (not yet classified; L-erythro = (3S, 4S) configuration), L- Fuculose-1-phosphataldolase ([EC 4.1.2.17]; D-erythro = (3R, 4R) configuration) or L-rhamnulose-1-phosphataldolase ([EC 4.1.2.19]; L-threo- = (3R, 4S) -config ration). This finding is highly surprising in that it is used here  The glycosylated aldehydes are sterically extraordinary due to the large glycoside residues are demanding. It was from numerous previous works with these Enzymes known (for SdT see e.g. the reviews by W.-D. Fessner et al. Top. Curr. Chem. 1996, 184, 97-194; H.J.M. Gijsen et al. Chem. Rev. 1996, 96, 443-473; C.-H. Wong et al. Appl. Chem. 1995, 107, 453-474) that the DHAP aldolases a have wide substrate tolerance for simple and substituted aliphatic aldehydes and therefore also good for the synthesis of monosaccharides and their derivatives are suitable (see, for example, PCT / WO-8 303 846, US-5 143 831, DE-41 11 971, US- 5 329 025, PCT / WO-9 516 049), but sterically hindered substrates such as wise pivalaldehyde are unable to implement.

Examples of the range of variation of the reaction products in question are in Scheme 4 listed.

It is advantageous for the process according to the invention that the additions are highly diastere run reoselectively and the oligosaccharides and their derivatives in high chemi have clean and optical purity. Since all four known types DHAP aldolases can be used optionally, are based on a common precursor in all corresponding possible diastereomers in predictably generated. It is also particularly advantageous that the enzymati aldol additions in an aqueous environment free of protective groups, especially mild ones Reaction conditions at room temperature and practically neutral pH (before preferably 6.8-7.0), which increases the stability of both the glycosidic Binding (s) as well as the reaction component DHAP with a sufficiently high reak speed remain guaranteed. The high tolerance of the enzyme reaction can also advantageously be used for synthesis within the scope of the method according to the invention higher oligosaccharides (tri, tetra, etc.) can be used, as well as for synthesis chemically modified (typically substituted, alkylated, acylated, acetali fixed) oligosaccharides with a modified activity profile.

Examples of the range of variation of the modifications in question are in Scheme 5 listed.  

Each of the aldolases listed above preferably uses DHAP as the nucleophi len aldol donor, but also a number of structurally related ones Donors are used. The third key step after the invention However, the process can alternatively also be carried out by other aldolases with different sub strat specificity are catalyzed, for example, instead of DHAP or pyruvate Use phosphoenol pyruvate as the preferred donor substrate and thereby from Sugar-derived 2-keto acids with an interesting biological activity profile produce. Other aldolases use acetaldehyde or glycine to synthesize Aldoses or a-amino-b-hydroxy acids. A list of preparative in the sense of the Erfin usable aldolases and the structural variants available with them are a known from the literature (for SdT see e.g. the reviews by H. J. M. Gijsen et al. Chem. Rev. 1996, 96, 443-473; C.-H. Wong et al. Angew Chem. 1995, 107, 453-474).

Because of the breadth of applicability for each key step, the flexibility and reliability of the entire process as well as its rapid and protective group pen-free feasibility, the method according to the invention can also be used for Creation of combinatorial oligosaccharide libraries.

The DHAP used for the synthesis is commercially available or can be economical mix z. B. by glycerol phosphate oxidase-mediated oxidation of commercial Glycerol phosphate (see. DE-43 04 097.7) are generated. The phosphorylated pro Products are based on standard processes such as ion exchange processes easy to isolate, while after phosphate ester hydrolysis (e.g. catalyzed by Phosphatases) the free oligosaccharides are obtained.

DHAP aldolases with different diastereospecificity and from different Organisms are commercially available (e.g. Boehringer Mannheim GmbH, Sand hofer Str. 116, 68298 Mannheim, DE; Sigma Chemie GmbH, Grünwalder Weg 30, 82039 Deisenhofen, DE). In the context of the present invention, in particular which uses the following enzyme preparations from Boehringer Mannheim: aldolases  from rabbit muscle, Staphylococcus carnosus and Escherichia coli, glycerin phosphate oxidase from microorganisms. L-glycerol phosphate for the production of DHAP was purchased from Genzyme (Haverhill, UK).

The compounds of formula (I) according to the invention are useful for inhibition and prevention of cell adhesion and diseases mediated by cell adhesion. she can be used to treat inflammatory diseases, asthma, dermatitis, Rheumatoid arthritis, osteoarthritis, multiple sclerosis, malignant diseases, Autoimmune diseases and bacterial, fungal and viral infections Exciters are used.

The compounds according to the invention are valuable precursors or constituents for active pharmaceutical ingredients, in particular for the treatment of the abovementioned Diseases.  

Scheme 2

Variety of glycosides

Scheme 3

Variety of chemical chain extensions

Scheme 4

Range of variation of the enzymatic chain extension

Scheme 5

Variety of derivatizations

The following examples are intended to illustrate the various aspects of the invention make it clear without limiting the invention.

Output connections General Working Instructions for Glycosidation 1 (AAV 1)

A 0.5 M solution of the monosaccharide in alcohol is heated under reflux for 1.5 h with an acidic ion exchanger (100 g per mol monosaccharide). The ion exchanger is filtered off and excess alcohol is removed in vacuo. The glycosid is purified by recrystallization or chromatography on silica gel.

• 1. Allyl-α-D-glucopyranoside (1a).
  According to AAV 1, from glucose and allyl alcohol, 15.0 g (49%).
• 2. Allyl-α-D-galactopyranoside (2a).
  According to AAV 1, from galactose and allyl alcohol, 34 g (28%).
• 3. Allyl-2-acetamido-2-deoxy-α-D-glucopyranoside (3a).
  According to AAV 1, from N-acetylglucosamine and allyl alcohol, 36.5 g (28%).
  Boron trifluoride etherate is used instead of an acidic ion exchanger.
• 4. Allyl-α-D-mannopyranoside (4a).
  According to AAV 1, from mannose and allyl alcohol, 10.6 g (62%).
• 5. Allyl-β-L-fucopyranoside (7b).
  According to AAV 1, from fucose and allyl alcohol, 5 g (30%).
• 6. Allyl-α-L-rhamnopyranoside (8a).
  According to AAV 1, from rhamnose and allyl alcohol, 15 g (84%).
• 7. Allyl-α-D-xylopyranoside (9a).
  According to AAV 1, from D-xylose and allyl alcohol, 45 g (52%).
• 8. Allyl-β-D-arabinopyranoside (10b).
  According to AAV 1, from D-arabinose and allyl alcohol, 23 g (24%).
• 9. Allyl-β-D-ribopyranoside (11b).
  According to AAV 1, from D-ribose and allyl alcohol, 16.7 g (44%).
• 10. Allyl-2,3-O-isopropylidene-β-D-ribofuranoside (16b).
  According to AAV 1, from D-ribose, acetone and allyl alcohol, 29.6 g (65%).

General Working Instructions for Glycosidation 2 (AAV 2)

1 equivalent of the peracetylated halogenose of the desired monosaccharide is reacted with 1.1 equivalents of mercury (II) cyanide and the desired alcohol (1 equivalent) in acetonitrile (0.2 M). After the reaction is complete, the solvent and excess alcohol are removed in vacuo and the residue is taken up in chloroform. The organic phase is washed thoroughly with saturated sodium chloride solution, dried over sodium sulfate and concentrated in vacuo. The peracetylated glycoside is further purified if necessary by recrystallization or chromatography on silica gel.

• 11. Allyl-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside (1b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and allyl alcohol (as solvent), 12.6 g (81%).
• 12. Allyl-2,3,4,6-tetra-O-acetyl-β-D-galactopyranoside (2b-Ac ₄ ).
  According to AAV 2, from acetobromogalactose and allyl alcohol, 40.8 g (78%).
• 13. Allyl-2-acetamido-3,4,6, -tri-O-acetyl-2-deoxy-β-D-glucopyranoside (3b-Ac ₃ ).
  According to AAV 2, from acetochlorglucosamine and allyl alcohol, 245 g (68%).
• 14. 1-O- (Methyl-2,3,4-tri-O-acetyl-β-D-glucopyranuronosyl) -2-propen-1-ol (5b-Ac ₃ ).
  According to AAV 2, from methyl 2,3,4-tri-O-acetyl-aD-glucopyranuronosyl bromide and allyl alcohol, 19.3 g (86%)
• 15. 1-O- (Methyl-2,3,4,6-tetra-O-acetyl-β-D-galactopyranuronosyl) -2-propen-1-ol (6b-Ac ₃ ).
  According to AAV 2, from methyl 2,3,4-tri-O-acetyl-aD-galactopyranuronosyl bromide and allyl alcohol, 5.08 g (65%).
• 16. Allyl-2,3,4-tri-O-acetyl-β-D-xylopyranoside (9b-Ac ₃ ).
  According to AAV 2, from acetobromxylose and allyl alcohol, 8.1 g (85%).
• 17.Allyl-2,3,6-tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) - β-D-glucopyranoside (12b- Ac ₇ ).
  According to AAV 2, from acetobromcellobiose and allyl alcohol, 20.0 g (98%).
• 18.Allyl-2,3,6-tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -β-D-glucopyranoside (13b- Ac ₇ ).
  According to AAV 2, from acetobromolactose and allyl alcohol, 12.9 g (66%).
• 19.Allyl-2,3,6-tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl) - β-D-glucopyranoside (14b- Ac ₇ ).
  According to AAV 2, from acetobromomaltose and allyl alcohol, 11.6 g (34%).
• 20. Allyl-2,3,5-tri-O-acetyl-β-D-ribofuranoside (15b-Ac ₃ ).
  According to AAV 2, from acetochlorribofuranose and allyl alcohol, 1.64 g (16%).
• 21. 1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -2-methyl-2-propen-1-ol (17b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and methallyl alcohol, 41.9 g (90%).
• 22. 1-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -2-methyl-2-propen-1-ol (18b-Ac ₄ ).
  According to AAV 2, from acetobromgalactose and methallyl alcohol, 7.2 g (90%).
• 23. 1-O- (2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl) -2-methyl-2-propen-1-ol (19b-Ac ₃ ).
  According to AAV 2, from acetochlorglucosamine and methallyl alcohol, 6.79 g (85%).
• 24. 1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -1-hydroxypropanone (20b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and hydroxyacetone, 3.75 g (98%).
• 25. 1-O- (2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl) -1-hydroxypropanone (21b-Ac ₃ ).
  According to AAV 2, from acetochlorglucosamine and hydroxyacetone, 2.62 g (65%).
• 26. 4-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -4-hydroxy-2-butanone (22b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and 4-hydroxy-2-butanone, 18.5 g (39%).
• 27. 1-O- (2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl) -3-methyl-2-buten-1-ol (23b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and isoprenol, 7.21 g (87%).
• 28. (E) -1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -3-phenyl-2-propen-1-ol (24b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and cinnamon alcohol, 3.41 g (74%).
• 29. 2 - ((2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl) oxymethyl) propenoic acid ethyl ester (25b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and ethyl 2- (hydroxymethyl) propenate, 4.05 g (88%).
• 30. (2RS) -2-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -1-O-trityl-3-buten-1,2-diol (26b- Ac ₄ Tr).
  According to AAV 2, from acetobromglucose and 1-O-trityl-3-buten-1,2-diol, 1.57 g (24%); Mixture of the two diastereomers in a ratio of 4: 3 (de = 14%).
• 31. (2RS) -2-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -3-buten-1,2-diol (27b-Ac ₄ Tr).
  According to AAV 2, from acetobromo galactose and 1-O-trityl-3-buten-1,2-diol, 417 mg (63%); Mixture of the two diastereomers in a ratio of 4: 3 (de = 15%).
• 32. (2RS) -2-O- (methyl-2,3,4-tri-O-acetyl-β-D-glucopyranuronosyl) -3-butene-1,2-diol (28b-Ac ₃ ).
  According to AAV 2, from methyl (2,3,4-tri-O-acetyl-aD-glucopyranuronosyl) bromide and 1- O-trityl-3-buten-1,2-diol, 4.8 g (37%); almost equimolar mixture of the two diastereomers (de = 3%).
• 33. (2RS) -2-O- (2,3,6-tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl) -bD -glucopyranosyl) -3-buten-1,2-diol (29b-Ac ₇ ).
  According to AAV 2, from acetobromomaltose and 1-O-trityl-3-buten-1,2-diol, 3.2 g (26%); Mixture of the two diastereomers in a ratio of 3: 2 (de = 20%).
• 34. (2S) -1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -3-buten-1,2-diol (30b-Ac ₄ ).
  According to AAV 2, from acetobromglucose and (2S) -3-butene-1,2-diol (1.1 equivalents), 1.73 g (41%).
• 35. (2RS) -1,2-Di-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -3-buten-1,2-diol (31b-Ac ₈ ).
  According to AAV 2, from acetobromglucose (1.5 equivalents) and (2RS) -3-butene-1,2-diol (1 equivalent), 3.1 g (41%).
• 36. (2S) -1,2-Di-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -3-buten-1,2-diol (32b-Ac ₈ ).
  According to AAV 2, from acetobromglucose (1.5 equivalents) and (2S) -3-butene-1,2-diol (1 equivalent), 3.4 g (45%).
• 37. 4-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -4-hydroxycyclohexanone (33b-Ac ₄ ).
  According to AAV 2, from acetobromglucose (1 equivalent) and 4-hydroxycyclohexanone (1 equivalent), 1.62 g (15%).
• 38. 2-Hydroxy-4-phenyl-2-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -3-butenaldimethylacetal (34b-Ac ₄ ).
  According to AAV 2, from acetobromglucose (1 equivalent) and 2-hydroxy-4-phenyl-3-butenaldimethyl acetal (1 equivalent), 3.04 g (60%).
• 39. 2-Hydroxy-4-phenyl-2-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -3-butenaldimethyl acetal (35b-Ac ₄ ).
  According to AAV 2, from acetobromgalactose (1 equivalent) and 2-hydroxy-4-phenyl-3-butenaldimethyl acetal (1 equivalent), 4.03 g (75%).
• 40. 2-Azido-4-phenyl-1-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -3-buten-1-ol (36b-Ac ₄ ).
  According to AAV 2, from acetobromo galactose (1 equivalent) and 2-azido-4-phenyl-3-buten-1-ol (1 equivalent), 4.6 g (89%).
• 41. 1,3-bis-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -2-methylene propane-1,3-diol (37b-Ac ₈ ).
  According to AAV 2, from acetobromglucose and 2-methylene propane-1,3-diol, 12.43 g (37%).
• 42. 1,1-bis - ((2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) oxymethyl) -3-buten-1-ol (38b-Ac ₈ ).
  According to AAV 2, from acetobromgalactose and 2-allylglycerol, 1.46 g (31%).
• 43. (4RS) -4-allyl-2,2-dimethyl-4 - ((2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -oxymethyl) -1,3-dioxolane ( 39b-Ac ₄ ).
  According to AAV 2, from acetobromglucose (1 equivalent) and (4RS) -4-allyl-2,2-dimethyl-4- (hydroxymethyl) -1,3-dioxolane (1 equivalent), 4.22 g (84%).

General working procedure for S-glycosylation (AAV 3)

One equivalent of thiourea is added to a 1 M solution of the corresponding acetobromosugar in acetone. The solution is heated to boiling for 15 minutes until the resulting isothiuronium bromide begins to precipitate. The mixture is allowed to cool, the solid is filtered off with suction and washed with a little acetone. 25 mmol of the isothiuronium bromide are dissolved in a solution of 5 g of sodium pyrosulfite in 20 ml of water and underlayered with 25 ml of methylene chloride. With stirring, the mixture is refluxed for 20 minutes. After cooling, the organic phase is washed out with water, dried over sodium sulfate and concentrated. The same volume of 1 MK ₂ CO ₃ solution and 1.1 equivalents of electrophile is added to a 1 M solution of b-glycosyl mercaptan in acetone. The mixture is stirred for 4 hours at room temperature. The acetone is removed in vacuo and the aqueous solution extracted 3 times with ethyl acetate. The combined organic phases are saturated with. Washed NaCl solution, dried over sodium sulfate and concentrated. Chromatograph on silica gel for purification.

• 44. S- (2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl) allylthiol (47b-Ac ₄ ).
  According to AAV 3, from acetobromglucose and allyl bromide, 2.3 g (67%).
• 45. (2RS) -3-S- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -2-hydroxy-3-mercaptopropanal diethylacetal (48b-Ac ₄ ).
  According to AAV 3, from acetobromglucose and glycidaldehyde diethylacetal, 3.5 g (96%).
• 46. (2RS) -3-S- (2,3,4-triacetyl-β-L-fucopyranosyl) -2-hydroxy-3-mercaptopropanal-diethyl acetal (49b-Ac ₄ ).
  According to AAV 3, from acetobrornfucose and glycidaldehyde diethylacetal, 1.6 g (91%).

General working procedure for N-glycosylation (AAV 4)

A 0.5 M solution of the corresponding sugar in allylamine is stirred under a nitrogen atmosphere for 2 to 3 days. Excess allylamine is removed in vacuo and the residue is taken up 3 times in anhydrous pyridine and concentrated. After dissolving in anhydrous pyridine (3 ml / mmol sugar), cooling to 0 ° C. and addition of acetic anhydride (1.5 ml / mmol sugar), the mixture is left to stand overnight. After narrowing i. V. and taking up in ethyl acetate (10 ml / mmol) is washed twice with sat. NaHCO ₃ solution and with sat. Washed NaCl solution and the organic phase dried over sodium sulfate. After filtration over silica gel (ethyl acetate), the residue is recrystallized from methyl tert-butyl ether and a little ethyl acetate.

• 47. N-Acetyl-N- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) allylamine (44b-Ac ₄ ).
  According to AAV 4, from glucose, 4.5 g (80%).
• 48. N-Acetyl-N- (2,3,4-tri-O-acetyl-β-D-fucopyranosyl) allylamine (45b-Ac ₃ ).
  According to AAV 4, from fucose, 2.3 g (67%).

General working procedure for C-glycosylation (AAV 5)

In a heated flask, 1.5 to 2 equivalents of allyltrimethyl silane and then slowly 0.1 equivalents of trimethylsilyl triflate are added to a 0.5 to 1 M solution of the acetylated sugar in dry nitromethane at 0 ° C. The reaction is kept at 0 ° C. for up to 8 h and up to three times 0.1 equivalents are added during this time until the reaction is complete after TLC control (CH / EE 3: 2). You pour carefully on ice-cold sat. NaHCO ₃ solution (4 ml / ml nitromethane), stirred until the trimethylsilyl triflate is neutralized and the aqueous phase is extracted 5 times with ethyl acetate (0.8 ml / ml nitromethane). The combined organic phases are saturated with sat. Washed NaCl solution and dried over sodium sulfate. After concentration, the mixture is filtered through silica gel or chromatographed to separate the diastereomers.

• 49. 3- (2,3,4-Tri-O-acetyl-α-D-xylopyranosyl) -1-propene (40a-Ac ₃ ) and 3- (2,3,4-Tri-O-acetyl-β -D-xylopyranosyl) -1-propene (40b-Ac ₃ ).
  According to AAV 5, from xylose tetraacetate, 1.6 g (43%) 40a-Ac ₃ , 1.2 g (32%) 40b-Ac ₃ .
• 50. 3- (2,3,4-Tri-O-acetyl-α-D-arabinopyranosyl) -1-propene (41a-Ac ₃ ) and 3- (2,3,4-Tri-O-acetyl-β -D-arabinopyranosyl) -1-propene (41b-Ac ₃ ).
  According to AAV 5, from D-arabinosetetraacetate, 1.4 g (70%) mixture of 41a-Ac ₃ and 41b-Ac ₃ .
• 51. 3- (2,3,4-Tri-O-acetyl-α-L-arabinopyranosyl) -1-propene (42a-Ac ₃ ) and 3- (2,3,4-Tri-O-acetyl-β -L-arabinopyranosyl) -1-propene (42b-Ac ₃ ).
  According to AAV 5, from L-arabinosetetraacetate, 1.3 g (65%) mixture of 42a-Ac ₃ and 42b-Ac ₃ .
• 52. 3- (2,3,4-Tri-O-acetyl-α-L-rhamnopyranosyl) -1-propene (43a-Ac ₃ ).
  According to AAV 5, from rhamnose tetraacetate, 14.6 g (60%).

General procedure for peracetylation (AAV 6)

A 0.4 M solution of the unprotected saccharide in acetic anhydride is mixed with 1 ‰ (w / v) sodium acetate and heated to reflux for 2 h. The solution is poured onto 2 volumes of ice water and neutralized with solid sodium hydrogen carbonate. It is extracted three times with 2 volumes of chloroform. The combined organic phases are washed thoroughly with sodium bicarbonate and brine, dried over sodium sulfate and the solvent removed in vacuo. The peracetate obtained is further purified if necessary by recrystallization or chromatography.

• 53. 2,3,4,6-tetra-O-acetyl-α-allyl-D-glucopyranoside (1a-Ac ₄ ).
  According to AAV 6, from 1a, 14.3 g (97%).

General working instructions for ozonolysis (AAV 7)

In a 10-500 mM solution of the saccharide in methanol (approx. 1% water content; with low solubility addition of ethyl acetate or chloroform), ozone is introduced at -78 ° C until a blue color persists. Excess ozone is expelled by introducing air. Dimethyl sulfide is added dropwise with stirring (0.5 ml per 1 mmol olefin). The solution is slowly thawed and left to stand at room temperature until a test with AI starch paper is negative (4-16 h), then concentrated in vacuo. The remaining syrup is dissolved several times in methanol (5-10% water content) (with low solubility, add ethyl acetate or chloroform) and concentrated again in vacuo. The carbonyl compounds contain changing proportions of dimethyl sulfoxide and are used for further chemical reactions without purification.

• 54. 2-O- (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl) -glykolaldehyd (1a-Ac _4-0).
  According to AAV 7, from 1a-Ac ₄ (14.1 g). The product is a mixture of free aldehyde and two diastereomeric methyl hemiacetals in a ratio of 1: 2: 2.
• 55. 2-O- (2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl) glycol aldehyde (1b-Ac ₄ -O).
  According to AAV 7, from 1b-Ac ₄ (13.2 g). The product is a 6: 1: 1 mixture of free aldehyde and two diastereomeric methyl hemiacetals.
• 56. 2-O-α-D-galactopyranosylglycolaldehyde (2a-O).
  According to AAV 7, from 2a (4.41 g).
• 57. 2-O- (2,3,4-Tri-O-acetyl-β-D-xylopyranosyl) glycol aldehyde (9b-Ac ₃ -O).
  According to AAV 7, from 9b-Ac ₃ (10.5 g). The product is present as a mixture of free aldehyde and two diastereomeric methyl hemiacetals in approximately equal proportions.
• 58. 2-O- (2,3,6-Tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) - β-D-glucopyranosyl ) -glycolaldehyde (12b-Ac ₇ -O).
  According to AAV 7, from 12b-Ac ₇ (18.9 g). The product is present as a mixture of two diastereomeric methyl hemiacetals in approximately equal proportions and about 10% free aldehyde.
• 59. 2-O- (2,3,6-Tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-bD-galactopyranosyl) -β-D-glucopyranosyl) - glycolaldehyde (13b-Ac ₇ -O).
  According to AAV 7, from 13b-Ac ₇ (9.68 g). The product is present as a mixture of two diastereomeric methyl hemiacetals in approximately equal proportions and about 25% free aldehyde.
• 60. 2-O- (2,3-O-isopropylidene-β-D-ribofuranosyl) glycol aldehyde (16b-O).
  According to AAV 7, from 16b (4.6 g).
• 61. 1-O- (2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl) -1-hydroxypropanone (17b-Ac ₄ -O).
  According to AAV 7, from 17b-Ac ₄ (20.1 g). The product corresponds to the compound 20b-Ac ₄ produced by glucosylation of hydroxyacetone.
• 62. 1-O- (2,3,4,6-Tetra-O-acetyl-β-D-galactopyranosyl) -1-hydroxypropanone (18b-Ac ₄ -O).
  According to AAV 7, from 18b-Ac ₄ (8.3 g).
• 63. 1-O- (2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl) -1-hydroxypropanone (19b-Ac ₃ -O).
  According to AAV 7, from 19b-Ac ₃ (401 mg). The product corresponds to the compound 21b-Ac ₃ produced by glycosylation of hydroxyacetone.
• 64. Ethyl 3- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -3-hydroxy-2-oxopropane (25b-Ac ₄ -O).
  According to AAV 7, from 25b-Ac ₄ (3.4 g). The product is a mixture of free keto compound and two diastereomeric methyl half ketals in a ratio of 2: 2: 1.
• 65. 1,3-bis-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -1,3-dihydroxy propanone (37b-Ac ₈ -O).
  According to AAV 7, from 37b-Ac ₈ (12.4 g).

Manufacturing examples General working instructions for allylation 1 (tin / indium) (AAV 8)

A 0.1 M solution of the unprotected aldehyde or ketone in an ethanol / water (5: 1) or ethanol / acetate buffer (2 M, pH 4.8) (5: 1) mixture is mixed with tin or indium powder (2 equivalents) and allyl bromide (3-4 equivalents) were added. It is irradiated with ultrasound until conversion is complete. The reaction mixture is sat. NaHCO ₃ solution. neutralized and centrifuged off from tin salts (if necessary, further tin salts can be separated off by adding phosphate buffer (1 M, pH 7.0-7.5) and centrifuging again). The clear supernatant is i. Vac. concentrated and the remaining syrup chromatographed on silica gel.

• 66. (2RS) -1-O-α-D-galactopyranosyl-4-pentene-1,2-diol (52a).
  According to AAV 8, from 2a-O, 5.1 g (96%). The product corresponds to that produced according to AAV 6, AAV 7, AAV 9 and AAV 10 from 2a.
  ¹³ C-NMR (D ₂ O, 125 MHz): d = 137.35 / 137.10 (C-4), 120.66 / 120.54 (C-5), 101.39 / 100.96 (C-1 '), 74.03 / 73.59 (C-1 ), 73.59 (C-5 '), 72.42 / 72.06 (C-2), 72.14 /72.13 (C-4'), 71.97 / 71.95 (C-3 '), 71.16 / 71.13 (C-2'), 63.89 /63.84 (C-6 '), 39.86 /39.84 (C-3).
• 67. (2RS) -1-O- (2-acetamido-2-deoxy-α-D-glucopyranosyl) -4-pentene-1,2-diol (53a).
  According to AAV 7 and AAV 8, from 3a, 6.6 g (80% for 2 steps).
  13 C-NMR (D ₂ O, 75 MHz): d = 177.04 / 176.99 (Ac), 137.14 / 137.01 (C-4), 120.56 (C-5), 100.14 / 99.79 (C-1 '), 74.61 ( C-5 '), 73.81 / 73.71 (C-3'), 73.65 / 73.40 (C-1), 72.66 (C-4 '), 72.36 / 72.05 (C-2), 63.21 (C-6'), 56.41 / 56.32 (C-2 '), 39.90 / 39.81 (C-3), 24.64 (Ac).
• 68. (2RS) -1-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl) -4-pentene-1,2-diol (53b).
  According to AAV 7 and AAV 8, from 3b-Ac ₃ , 8.2 g (77% for 2 steps).
  ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.52 (Ac), 137.21 / 137.03 (C-4), 120.73 /120.60 (C-5), 104.45 / 104.23 (C-1 '), 78.67 ( C-5 '), 76.56 (C-3'), 75.82 (C-1), 72.72 (C-4 '), 72.28 / 72.10 (C-2), 63.50 (C-6'), 58.39 (C- 2 '), 39.83 (C-3), 25.03 (Ac).
• 69. (2RS) -1-O-α-D-galactopyranosyl-3,4-pentadiene-1,2-diol (98a).
  A 100 mM solution of 2a-O (from 2a according to AAV 7) in ethanol / water = 5/1 or a 300 mM solution of 2a-Ac ₄ -O (from 2a-Ac ₄ according to AAV 7) in tetrahydro furan / Ethyl acetate = 1/1 is slightly acidified with 2N HCl (pH 3), tin powder (2 equivalents) and propargyl bromide (4-5 equivalents) are added and ultrasound is applied. After the end of the reaction (approx. 45 min, TLC control), NaHCO ₃ solution. neutralized and centrifuged from tin salts. The centrifugate is desalinated on ion exchangers (HCO ₃ ⁻ and H⁺ form) and after concentration i. Vac. chromatographed on silica gel (CHCl ₃ / MeOH). The peracetylated compound is deprotected according to AAV 10.
  from 2a: 4.9 g (75%) mixture of all and alkyne (55: 45)
  from 2a-Ac ₄ : 910 mg (87%) mixture of allen and alkyne (55: 45)
  ¹³ C-NMR (D ₂ O, 75 MHz): d = 210.51 (C-4), 101.65 / 101.17 (C-1 '), 92.92 / 92.74 (C-3), 80.38 / 80.35 (C-5), 73.81 (C-5 '), 73.10 / 72.74 (C-1), 72.25 (C-4'), 72.04 (C- 3 '), 71.24 (C-2'), 70.72 / 70.37 (C-2), 63.96 (C-6 ').

General working instructions for allylation 2 (zinc (AAV 9)

A 1 M solution of the peracetylated saccharide is mixed with 1 volume of THF and 4 volumes of saturated aqueous NH ₄ Cl solution. (In the case of solubility problems, up to four times the amount of ethyl acetate and THF can be used.) 2 to 4 equivalents of zinc dust are added and 2 to 4 equivalents of allyl bromide are added dropwise over 2 h with vigorous stirring. If the conversion is incomplete (TLC control), the mixture is stirred for a longer time and 1 equivalent of allyl bromide is slowly added. The inorganic precipitate is filtered off and washed thoroughly with diethyl ether and chloroform. The combined liquid phases are separated and the aqueous phase extracted three times with three times the volume of diethyl ether or chloroform (depending on the solubility of the saccharide). The combined organic phases are dried over sodium sulfate and concentrated in vacuo. The homoallylic alcohols are usually sufficiently pure (∼ 95%). Most of the DMSO residues from any previous ozonolysis are also removed during processing.

• 70. (2RS) -1-O- (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl) -4-pentene-1,2-diol (51a-Ac ₄ ).
  According to AAV 9, from 1a-Ac ₄ -O, 15.7 g (quant. Yield for 2 steps); almost equimolar mixture of the two diastereomers (de = 2%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.60, 170.14, 170.05, 170.00, 169.57, 169.55, 133.85 (C-4), 133.77 (C-4), 118.12 (CS (2x)), 96.64 ( C-1 '), 96.00 (C-1'), 72.82 (C- 1), 72.43 (C-1), 70.79, 70.13, 70.09, 69.53, 69.26, 68.53, 67.48, 61.93 (C-6 '), 61.88 (C-6 '), 37.97 (C-3), 37.75 (C-3), 20.70 (CH ₃ ), 20.67 (CH ₃ ), 20.64 (CH ₃ ), 20.60 (CH ₃ ).
• 71. (2RS) -1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -4-pentene-1,2-diol (51b-Ac ₄ ).
  According to AAV 9, from 1b-Ac ₄ -O, 12.7 g (86% for 2 steps); almost equimolar mixture of the two diastereomers (de = 5%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.67 (Ac (2x)), 170.23 (Ac (2x)), 169.52 (Ac (2x)), 169.45 (Ac (2x)), 134.08 (C- 4 (2x)), 117.83 (C-5), 117.79 (C-5), 101.40 (C- 1 '), 101.38 (C-1'), 74.98 (C-1), 74.41 (C-1), 72.70 (2 C), 71.87 (2 C), 71.40, 71.33, 69.71, 69.66, 68.41 (2 C), 61.97 (C-6 '(2x)), 37.64 (C-3), 37.59 (C-3) , 20.67 (CH ₃ ), 20.58 (CH ₃ ).
• 72. (2RS) -1-O- (2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl) -4-pentene-1,2-diol (54a-Ac ₄ ).
  According to AAV 6, AAV 7 and AAV 9, from 4a, 24.2 g (92% for 3 steps); almost equimolar mixture of the two diastereomers.
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.66 (Ac), 170.08 (Ac), 170.00 (Ac), 169.76 (Ac), 133.89 (C-4), 133.85 (C-4), 118.17 ( C-5 (2x)), 98.19 (C-1 '), 97.82 (C-1'), 72.31 (C-1 (2x)), 69.49 (2 C), 69.41, 69.37, 69.07 (2 C), 68.70 (2 C), 66.14, 66.10, 62.55 (C-6 '), 62.47 (C-6'), 38.00 (C-3), 37.94 (C-3), 20.86 (CH ₃ ), 20.73 (CH ₃ ), 20.70 (CH ₃ ).
• 73. (2RS) -1-O- (methyl-2,3,4-tri-O-acetyl-β-D-glucopyranuronosyl) -4-pentene-1,2-diol (55b-Ac ₃ ).
  According to AAV 7 and AAV 9, from 5b-Ac ₃ , 4. 12 g (82% for 2 steps); almost equimolar mixture of the two diastereomers (de <5%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.06 (C = O (2x)), 169.42 (C = O), 169.39 (C = O (2x)), 169.33 (C = O), 167.18 ( C = O), 167.13 (C = O), 134.05 (C-4), 133.99 (C-4), 117.85 (C-5), 117.81 (C-5), 101.24 (C-1 '), 101.20 ( C-1 '), 74.93 (C-1), 74.44 (C-1), 72.42, 72.33, 71.96, 71.93, 71.36, 71.29, 69.63, 69.61, 69.31, 69.23, 52.96 (6'- OCH ₃ ), 52.94 (6'-OCH ₃ ), 37.62 (C-3), 37.59 (C-3), 20.64 (CH ₃ (2x)), 20.59 (CH ₃ (2x)), 20.48 (CH ₃ (2x)).
• 74. (2RS) -1-O- (methyl-2,3,4-tri-O-acetyl-β-D-galactopyranuronosyl) -4-pentene-1,2-diol (56b-Ac ₃ ).
  According to AAV 7 and AAV 9, from 6b-Ac ₃ , 5.5 g (96% for 2 steps); almost equimolar mixture of the two diastereomers (de = 9%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.04 (C = O (2x)), 169.84 (C = O (2x)), 169.58 (C = O), 169.45 (C = O), 166.53 ( C = O), 166.46 (C = O), 134.27 (C-4), 134.21 (C-4), 117.65 (C-5 (2x)), 101.72 (C-1 '), 101.52 (C-1' ), 75.70 (C-1), 74.60 (C-1), 72.30 (2 C), 70.44 (2 C), 69.76, 69.70, 68.65, 68.56, 68.26, 68.19, 52.87 (6'-OCH ₃ ), 52.82 (6'-OCH ₃ ), 37.68 (C-3), 37.61 (C-3), 20.75 (CH ₃ (2x)), 20.57 (CH ₃ (4x)).
• 75. (2RS) -1-O- (2,3,4-tri-O-acetyl-α-D-xylopyranosyl) -4-pentene-1,2-diol (59a-Ac ₃ ).
  According to AAV 6, AAV 7 and AAV 9, from 9a, 10.1 g (88% for 3 steps); almost equimolar mixture of the two diastereomers.
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.12 (Ac), 169.97 (Ac), 133.95 (C-4), 133.84 (C-4), 118.05 (C-5 (2x)), 96.71 ( C-1 '), 96.04 (C-1'), 72.59 (C-1), 72.13 (C-1), 70.98 (2 C), 69.59, 69.55, 69.27 (2 C), 58.51 (C-5 ' (2x)), 37.92 (C-3), 37.68 (C-3), 20.76 (CH ₃ ), 20.70 (CH ₃ ).
• 76. (2RS) -1-O- (2,3,4-tri-O-acetyl-β-D-xylopyranosyl) -4-pentene-1,2-diol (59b-Ac ₃ ).
  According to AAV 9, from 9b-Ac ₃ -O, 8.02 g (84% for 2 steps); almost equimolar mixture of the two diastereomers (de = 3%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.11 (Ac (2x)), 169.88 (Ac (2x)), 169.62 (Ac), 169.55 (Ac), 134.01 (C-4 (2x)), 117.91 (CS (2x)), 101.33 (C-1 '), 101.08 (C- 1'), 73.89 (C-1), 73.63 (C-1), 71.50, 71.39, 71.03, 70.85, 69.61, 69.49, 68.86, 68.74, 62.24 (C-5 '), 62.12 (C-5'), 37.66 (C-3), 37.61 (C-3), 20.73 (CH ₃ ).
• 77. (2RS) -1-O- (2,3,4-tri-O-acetyl-β-D-arabinopyranosyl) -4-pentene-1,2-diol (60b-Ac ₃ ).
  According to AAV 6, AAV 7 and AAV 9, from 10b, 8.9 g (90% for 3 steps); almost equimolar mixture of the two diastereomers.
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.37 (Ac), 170.32 (Ac), 170.26 (Ac), 170.13 (Ac), 170.11 (Ac), 133.88 (C-4), 133.77 (C- 4), 118.08 (C-5 (2x)), 97.45 (C-1 '), 96.78 (C-1'), 72.67 (C-1), 72.24 (C-1), 69.62, 69.31, 69.02 (2 C), 68.36 (2 C), 67.18, 67.14, 60.61 (C-5 '), 60.59 (C-5'), 37.94 (C-3), 37.66 (C-3), 20.93 (CH ₃ ), 20.77 (CH ₃ ), 20.75 (CH ₃ ), 20.72 (CH ₃ ).
• 78. (2RS) -1-O- (2,3,6-tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -β -D-glucopyranosyl) -4-pentene-1,2-diol (62b-Ac ₇ ).
  According to AAV 9, from 12b-Ac ₇ -O, 20.2 g (quant. Yield for 2 steps); almost equimolar mixture of the two diastereomers (de <8%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.51 (Ac (2x)), 170.37 (Ac (2x)), 170.22 (Ac (2x)), 169.81 (Ac (2x)), 169.74 (Ac) , 169.68 (Ac), 169.34 (Ac (2x)), 169.10 (Ac (2x)), 134.12 (C-4), 134.07 (C-4), 117.76 (C-5), 117.73 (C-5), 101.34 (C-1 '), 101.25 (C-1'), 100.78 (C-1 '' (2x)), 76.54, 75.35 (C-1), 74.57 (C-1), 72.91, 72.79, 72.39, 71.94, 71.60, 69.63, 67.78, 61.89 (C-6 °), 61.56 (C-6 °), 37.63 (C-3), 37.55 (C-3), 20.79 (CH ₃ ), 20.76 (CH ₃ ), 20.68 (CH ₃ ), 20.64 (CH ₃ ), 20.52 (CH ₃ ).
• 79. (2RS) -1-O- (2,3,6-Tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -β -D-glucopyranosyl) -4-pentene-1,2-diol (63b-Ac ₇ ).
  According to AAV 9, from 13b-Ac ₇ -O, 9.26 g (90% for 2 steps); almost equimolar mixture of the two diastereomers (de <6%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.41 (Ac (2x)), 170.34 (Ac (2x)), 170.16 (Ac (2x)), 170.05 (Ac (2x)), 169.75 (Ac ( 3x)), 169.68 (Ac), 169.11 (Ac (2x)), 134.17 (C- 4), 134.11 (C-4), 117.71 (C-5), 117.68 (C-5), 101.27 (C-1 '), 101.19 (C-1'), 101.03 (C-1 '' (2x)), 76.34 (2 C), 75.35 (C-1), 74.57 (C-1), 72.72 (4 C), 71.72 , 71.66, 70.95 (2 C), 70.67 (2 C), 69.63 (2 C), 69.11 (2 C), 66.69 (2 C), 62.06 (C-6 '), 62.06 (C-6'), 60.88 (C-6 ''), 37.66 (C-3), 37.56 (C-3), 20.77 (CH ₃ ), 20.70 (CH ₃ ), 20.62 (CH ₃ ), 20.60 (CH ₃ ), 20.49 (CH ₃ ).
• 80. (2RS) -1-O- (2,3,6-tri-O-acetyl-4-O- (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl) -β -D-glucopyranosyl) -4-pentene-1,2-diol (64b-Ac ₇ ).
  According to AAV 7 and AAV 9, from 14b-Ac ₇ , 12.3 g (quant. Yield for 2 steps); almost equimolar mixture of the two diastereomers (de <6%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.50 (Ac (4x)), 170.44 (Ac (2x)), 170.17 (Ac (2x)), 169.90 (Ac (2x)), 169.71 (Ac) , 169.66 (Ac), 169.41 (Ac (2x)), 134.15 (C-4), 117.71 (C-5), 117.69 (C-5), 100.85 (C-1 '), 95.61 (C-1'' ), 75.23 (2 C), 74.83 (C-1), 74.32 (C-1), 72.87 (2 C), 72.19 (3 C), 72.15 (1 C), 70.00 (2 C), 69.58, 69.55, 69.30 (2 C), 68.56 (2 C), 68.05 (2 C), 62.78 (C-6 '), 61.57 (C-6'), 37.69 (C-3), 37.62 (C-3), 20.86 ( CH ₃ (2x)), 20.74 (CH ₃ (2x)), 20.64 (CH ₃ (2x)), 20.61 (CH ₃ (2x)), 20.55 (CH ₃ (6x)).
• 81. (2RS) -1-O- (2,3,5-tri-O-acetyl-β-D-ribofuranosyl) -4-pentene-1,2-diol (65b-Ac ₃ ).
  According to AAV 7 and AAV 9, from 15b-Ac ₃ , 0.80 g (50% for 2 steps); almost equimolar mixture of the two diastereomers (de approx. 2%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.69 (Ac), 170.64 (Ac), 169.72 (Ac (2x)), 169.66 (Ac (2x)), 134.09 (C-4), 134.06 (C -4), 117.79 (C-5 (2x)), 105.99 (C-1 '), 105.71 (C-1'), 78.61, 78.55, 74.77, 74.73, 73.00 (C-1), 72.82 (C-1 ), 71.15 (2 C), 69.63, 69.52, 64.12 (C-5 '), 64.08 (C-5'), 37.77 (C-3), 37.77 (C-3), 20.78 (CH ₃ ), 20.57 ( CH ₃ ), 20.50 (CH ₃ ).
• 82. (2RS) -1-O- (2,3-O-isopropylidene-β-D-ribofuranosyl) -4-pentene-1,2-diol (66b).
  According to AAV 7 and AAV 9, from 16b, 2.67 g (49% for 2 steps).
  ¹³ C-NMR (D ₂ O, 75 MHz): d = 137.01 / 136.96 (C-4), 120.60 (C-5), 115.68 110.97 / 110.47 (C-1 '), 89.67 / 89.59 (C-4' ), 87.29 / 87.24 (C-3 ') *, 83.84 / 83.81 (C- 2') *, 74.11 / 73.79 (C-1), 72.15 / 71.80 (C-2), 65.21 (C-5 '), 39.99 / 39.94 (C-3), 28.05 (CH ₃ ), 26.41 (CH ₃ ).
• 83. (2RS) -1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -2-methyl-4-pentene-1,2-diol (67b-Ac ₄ ).
  According to AAV 9, from 17b-Ac ₄ -O or 20b-Ac ₄ , 22.3 g (quant. Yield for 2 steps); almost equimolar mixture of the two diastereomers (de = 11%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.64 (Ac (2x)), 170.22 (Ac (2x)), 169.41 (Ac (2x)), 169.34 (Ac (2x)), 133.54 (C- 4), 133.45 (C-4), 118.56 (C-5 (2x)), 101.43 (C-1 '(2x)), 77.36 (C-2), 77.30 (C-2), 72.67 (2 C) , 71.84 (2 C), 71.62 (C-1), 71.55 (C-1), 71.40, 71.37, 68.43 (2 C), 61.93 (C-6 '(2x)), 43.36 (C-3 (2x) ), 23.74 (2-CH ₃ ), 23.26 (2-CH ₃ ), 20.70 (CH ₃ ), 20.65 (CH ₃ ), 20.59 (CH ₃ ).
• 84. (2RS) -1-O- (2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl) -2-methyl-4-pentene-1,2-diol (68b-Ac ₄ ).
  According to AAV 9, from 18b-Ac ₄ -O, 8.96 g (quant. Yield for 2 steps); almost equimolar mixture of the two diastereomers (de = 6%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.41 (Ac (2x)), 170.25 (Ac (2x)), 170.10 (Ac (2x)), 169.50 (Ac (2x)), 133.56 (C- 4), 133.47 (C-4), 118.56 (C-5 (2x)), 101.96 (C-1 '(2x)), 77.36 (C-1), 77.27 (C-1), 71.65 (C-2 ), 71.57 (C-2), 70.79 (4 C), 69.02, 68.99, 67.01 (2 C), 61.31 (C-6 '(2x)), 43.36 (C-3 (2x)), 23.77 (2- CH ₃ ), 23.72 (2-CH ₃ ), 20.78 (CH ₃ ), 20.67 (CH ₃ (2x)), 20.58 (CH ₃ ).
• 85. (2RS) -1-O- (2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl) -2-methyl-4-pentene-1,2- diol (69b-Ac ₃ ).
  According to AAV 9, from 19b-Ac ₃ -O or 21b-Ac ₃ , 890 mg (quant. Yield for 2 steps); almost equimolar mixture of the two diastereomers (de = 11%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 172.19 (Ac (2x)), 171.01 (Ac (2x)), 170.84 (Ac (2x)), 169.47 (Ac (2x)), 133.55 (C- 4 (2x), 118.58 (C-5 (2x)), 101.63 (C-1 '(2x)), 76.75 (C-2 (2x)), 72.46 (2 C), 72.24 (C-1 (2x) ), 71.76 (2 C), 68.78 (2 C), 62.24 (C-6 '(2x)), 54.61 (C-2' (2x)), 43.49 (C-3), 43.23 (C-3), 23.51 (2-CH ₃ ), 23.34 (CH ₃ ), 23.25 (2-CH ₃ ), 20.78 (CH ₃ ), 20.64 (CH ₃ ).
• 86. (3RS) -1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -3-methyl-5-hexene-1,3-diol (72b-Ac ₄ ).
  According to AAV 9, from 22b-Ac ₄ , 4.5 g (98%); almost equimolar mixture of the two diastereomers.
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.69 (Ac (2x)), 170.26 (Ac (2x)), 169.46 (Ac (2x)), 169.43 (Ac (2x)), 134.14 (C- 5), 134.06 (C-5), 118.49 (C-6), 118.40 (C-6), 100.63 (C-1 '(2x)), 72.74, 71.86, 71.54 (C-3), 71.52 (C- 3), 71.25, 68.43, 66.74 (C- 1), 66.69 (C-1), 61.89 (C-6 '(2x)), 46.91 (C-4), 46.83 (C-4), 39.81 (C- 2), 39.72 (C- 2), 26.78 (3-CH ₃ ), 26.75 (3-CH ₃ ), 20.72 (CH ₃ ), 20.65 (CH ₃ ), 20.58 (CH ₃ (2x)).
• 87. (2RS) -2 - ((2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl) oxymethyl) -2-hydroxy-4-pentenoic acid methyl ester (75b-Ac ₄ ).
  According to AAV 9, from 25b-Ac ₄ -O, 2.71 g (77%); almost equimolar mixture of the two diastereomers (de <5%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 173.74 (C-1), 173.26 (C-1), 170.66 (Ac (2x)), 170.25 (Ac), 170.19 (Ac), 169.47 (Ac) , 169.45 (Ac), 169.40 (Ac), 169.24 (Ac), 131.46 (C-4), 131.11 (C-4), 119.35 (C-5), 119.10 (C-5), 101.45 (C-1 ' ), 101.42 (C- 1 '), 78.08 (C-2), 77.31 (C-2), 75.78 (2-CH ₂ OR), 73.66 (2-CH ₂ OR), 72.74, 72.70, 71.89, 71.71, 71.17 (2 C), 68.45, 68.35, 62.35 (CH ₂ (Et)), 62.04 (CH ₂ (Et)), 61.87 (C-6 '), 61.82 (C-6'), 39.50 (C-3 ( 2x)), 20.72 (CH ₃ ), 20.69 (CH ₃ ), 20.66 (CH ₃ ), 20.59 (CH ₃ ), 14.29 (CH ₃ (Et)), 14.15 (CH ₃ (Et)).
• 88. (2RS) -2-O-acetyl-1-O- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -2-methyl-3-butene-1,2- diol (76b-Ac ₅ ).
  2 ml of 1 M vinylmagnesium bromide solution (2 mmol) are slowly added dropwise to a solution of 650 mg (1.6 mmol) of 17b-Ac ₄ -O or 20b-Ac ₄ in 40 ml of tetrahydrofuran at 0 ° C. After 2 h, 0.1 ml (10 mmol) of acetic acid are carefully added at 0 ° C, then 2 ml of acetic anhydride, 1 ml of pyridine and 0.1 g of 4- (dimethylamino) pyridine. After 8 h, volatile constituents are removed in vacuo, the residue is taken up in 50 ml of methylene chloride, shaken with dilute hydrochloric acid, sodium hydrogen carbonate solution and saturated sodium chloride solution, dried over sodium sulfate and concentrated in vacuo. Chromatography on silica gel (cyclohexane / - ethyl acetate = 3/2) gives 470 mg (0.99 mmol, 62%) 76b-Ac ₅ as a colorless syrup with a diastereomer ratio of 3: 1 (de = 50%; locants of the main diastereomer in NMR, for example, T. underlined).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 171.19 (Ac), 170.70 (Ac), 170.28 (Ac), 169.46 (Ac), 169.40 (Ac), 141.83 (C-3), 141.47 (C- 3 ), 114.04 (C- 4 ), 113.74 (C-4), 101.58 (C-1 '(2x)), 78.04 (C- 2 ), 77.79 (C-2), 72.72 (2 C), 72.59 ( C-1), 72.46 (C- 1 ), 71.94 (2 C), 71.40 (2 C), 68.47 (2 C), 62.01 (C-6 '(2x)), 24.35 ( 2 -CH ₃ ), 24.14 (2-CH ₃ ), 21.04 (CH ₃ ), 20.70 (CH ₃ ), 20.67 (CH ₃ ), 20.59 (CH ₃ ).
• 89. (2RS) -2-O-acetyl-1-O- (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl) -3-buten-1,2-diol (77a- Ac ₅ ).
  At 78 ° C., ozone is passed into a solution of 1.69 g (4.35 mmol) of 1a-Ac ₄ in 40 ml of methylene chloride until a blue color remains. Excess ozone is expelled by introducing air, 2.2 ml of dimethyl sulfide are added dropwise with stirring and after 30 min a solution of 1.15 g (4.4 mmol) of triphenylphosphine in 5 ml of methylene chloride is added dropwise. 10 ml of 1 M vinylmagnesium bromide solution (10 mmol) are then slowly added dropwise at 0.degree.
  After 2 h, 0.2 ml (20 mmol) of acetic acid are carefully added at 0 ° C, then 10 ml of acetyl chloride, 5 ml of pyridine and 0.2 g of 4- (dimethylamino) pyridine. After 4 h, volatile constituents are removed in vacuo, taken up in 100 ml of methylene chloride, shaken with dilute hydrochloric acid, sodium hydrogen carbonate solution and saturated sodium chloride solution, dried over sodium sulfate and concentrated in vacuo. Chromatography on silica gel (cyclohexane / ethyl acetate = 3/2) gives 1.57 g (3.41 mmol, 78%) of 77a-Ac ₅ as a colorless syrup with almost equal proportions of the two diastereomers.
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.52 (Ac), 170.12 (Ac), 170.01 (Ac), 169.93 (Ac), 169.83 (Ac), 169.56 (Ac), 132.76 (C-3) , 132.69 (C-3), 118.65 (C-4 (2x)), 96.09 (C-1 '), 95.59 (C-1'), 73.37, 72.65, 70.83, 70.68, 69.98 (2 C), 69.17 ( C-1), 68.84 (C-1), 68.50, 68.45, 67.53, 67.50, 61.79 (C-6 '(2x)), 21.01 (CH ₃ ), 20.94 (CH ₃ ), 20.66 (CH ₃ (4x) ), 20.61 (CH ₃ ), 20.58 (CH ₃ (3x)).
• 90. 1,1-bis - ((2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) oxymethyl) -3-buten-1-ol (87b-Ac ₈ ).
  According to AAV 9, from 37b-Ac ₈ -O, 9.14 g (71%).
  ¹³ C-NMR (75.4 MHz; CDCl ₃ ): d = 170.66 (Ac), 170.60 (Ac), 170.20 (Ac), 170.13 (Ac), 169.42 (Ac (3x)), 169.20 (Ac), 132.59 (C -3), 118.68 (C-4), 101.51 (C-1 '), 101.20 (C-1'), 74.10 (C-1), 72.81 (1-CH ₂ OR), 72.68 (1-CH ₂ OR ), 72.68, 72.44, 71.93, 71.70, 71.51, 71.22, 68.46, 68.40, 61.87 (C-6 '(2x)), 38.65 (C-2), 20.72 (CH ₃ ), 20.66 (CH ₃ , (3x) ), 20.59 (CH ₃ , (2x)), 20.57 (CH ₃ , (2x)).
• 91. 1- (2,3,4-Tri-O-acetyl-α-D-xylopyranosyl) -4-penten-2-ol (90a-Ac ₃ ).
  According to AAV 7 and AAV 9, from 40a-Ac ₃ , 0.9 g (80%).
  ¹³ C NMR (CDCl ₃ , 75 MHz): d = 170.00 (Ac), 169.91 (Ac), 168.74 (Ac), 168.62 (Ac), 134.47 (C-2), 134.45 (C-2), 118.33 ( C-1), 117.90 (C-1), 74.14 (C-1 '), 71.08 (2 C), 68.86, 67.85, 67.71, 66.76, 66.62, 66.46, 66.30, 66.10, 65.98, 65.98, 65.73, 42.53 ( CH ₂ ), 41.90 (CH ₂ ), 37.38 (CH ₂ ), 36.49 (CH ₂ ), 21.08 (CH ₃ ), 20.87 (CH ₃ ), 20.84 (CH ₃ ), 20.78 (CH ₃ ), 20.50 (CH ₃ ).
• 92. 1- (2,3,4-Tri-O-acetyl-β-D-xylopyranosyl) -4-penten-2-ol (90b-Ac ₃ ).
  According to AAV 7 and AAV 9, from 40b-Ac ₃ , 0.8 g (78%).
  ¹³ C NMR (CDCl ₃ , 75 MHz): d = 170.83 (Ac), 170.39 (Ac), 170.24 (Ac), 134.96 (C-2), 118.48 (C-1), 118.15 (C-1 ), 78.67 (C-1 '), 75.74, 74.30, 74.09, 74.01, 72.41, 69.85, 69.53, 69.49, 67.05, 66.94, 42.68 (CH ₂ ), 41.90 (CH ₂ ), 38.38 (CH ₂ ), 38.02 ( CH ₂ ), 21.10 (CH ₃ ).
• 93. 1- (2,3,4-Tri-O-acetyl-α-L-rhamnopyranosyl) -4-penten-2-ol (93a-Ac ₃ ).
  According to AAV 7 and AAV 9, from 43a-Ac ₃ , 11.7 g (79%).
  ¹³ C NMR (CDCl ₃ , 75 MHz): d = 170.92 (CO ₂ ), 170.71 (CO ₂ ), 170.67 (CO ₂ ), 170.46 (CO ₂ ), 134.74 (C-2), 119.11 / 118.77 (C -1), 74.67 (C-1 '), 72.13 / 71.82 (C-2) *, 71.72 / 71.46 (C-3) *, 69.65 / 69.51 (C-4) *, 69.3 / 69.05 (C-5) *, 67.38 (C-4), 42.76 / 42.13 (C-5), 35.76 / 35.23 (C-3), 21.52 (CH ₃ ), 21.41 (CH ₃ ), 21.30 (CH ₃ ), 18.09 / 18.01 (C -6 ').
• 94. (4R, S) -N-acetyl-N- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -1-amino-4-penten-2-ol (94b- Ac ₄ ).
  According to AAV 7 and AAV 9, from 44b-Ac ₄ , 2.5 g (92%).
  13 C NMR (CDCl ₃ , 75 MHz): d = 173.66 (Ac), 172.76 (Ac), 172.32 (Ac), 171.79 (Ac), 170.69 (Ac), 170.54 (Ac), 170.09 (Ac), 169.84 (Ac), 169.56 (Ac), 168.98 (Ac), 134.57 (C-2), 134.47 (C-2), 134.00 (C-2), 133.81 (C-2), 130.91 (C-2), 128.81 (C-2), 118.31 (C-1), 118.04 (C-1), 117.51 (C-1), 117.37 (C-1), 80.64 (C-1 '), 79.73 (C-1'), 74.15, 73.56, 73.21, 73.06, 69.84, 69.54, 68.80, 68.06, 67.74, 61.53, 61.37, 61.20, 40.07, 39.50, 39.42, 26.98 (CH ₃ ), 23.75 (CH ₃ ), 22.87 (CH ₃ ), 22.50 ( CH ₃ ), 22.05 (CH ₃ ), 21.43 (CH ₃ ), 20.53 (CH ₃ ).
• 95. (2RS) -1-O-α-D-galactopyranosyl-3,3-dimethyl-4-pentene-1,2-diol (99a).
  According to AAV 7, AAV 9 (5 equivalents of 3,3-dimethylallyl bromide instead of allyl bromide) and AAV 10, from 2a-Ac ₄ , 950 mg (81%).
  ¹³ C NMR (D ₂ O, 75 MHz): d = 147.80 / 147.64 (C-4), 115.66 / 115.43 (C-5), 101.56 / 100.96 (C-1 '), 80.07 / 79.25 (C-2 ), 73.85 / 73.66 (C-5 '), 72.28 / 72.23 (C- 4'), 72.05 (C-3 '), 71.97 / 71.74 (C-1), 71.25 / 71.11 (C-2'), 63.96 /63.86 (C-6 '), 42.50 / 42.37 (C-3), 25.72 / 25.15 (CH ₃ ), 25.63 / 24.77 (CH ₃ ).

General Working Instructions for Deprotection (AAV 10)

Removal of acyl and trityl groups: A 0.1 M solution of the acylated compound in methanol is mixed with sodium methoxide (50 mg / 100 ml). After the reaction has ended (0.5-4 h), acidic ion exchanger (2 g / 100 ml) is added, stirred, suction filtered after 5 min and washed with methanol. Volatile components are removed in vacuo and the saccharide is further purified if necessary by crystallization or chromatography.

• 96. Allyl-β-D-glucopyranoside (1b).
  According to AAV 10, from 1b-Ac ₄ , 7.14 g (quant. Yield).
• 97. Allyl-β-D-galactopyranoside (2b).
  According to AAV 10, from 2b-Ac ₄ , 5.90 g (quant. Yield).
• 98. Allyl-2-acetamido-2-deoxy-β-D-glucopyranoside (3b).
  According to AAV 10, from 3b-Ac ₄ , 9.22 g (quant. Yield).
• 99. 1-O- (Methyl-β-D-glucopyranuronosyl) -2-propen-1-ol (5b).
  According to AAV 10, from 5b-Ac ₃ , 248 mg (quant. Yield).
• 100. 1-O-β-D-glucopyranosyl-3-methyl-2-buten-1-ol (23b).
  According to AAV 10, from 23b-Ac ₄ , 596 mg (quant. Yield).
• 101. (E) -1-O-β-D-glucopyranosyl-3-phenyl-2-propen-1-ol (24b).
  According to AAV 10, from 24b-Ac ₄ , 1.04 g (quant. Yield).
• 102. (2RS) -3-S-β-D-glucopyranosyl-2-hydroxy-3-mercaptopropanal-diethyl acetal (48b).
  According to AAV 10, from 48b-Ac ₄ , 1.4 g (98%).
• 103. (2RS) -1-O-α-D-glucopyranosyl-4-pentene-1,2-diol (51a).
  According to AAV 10, from 51a-Ac ₄ , 2.64 g (quant. Yield).
• 104. (2RS) -1-O-β-D-glucopyranosyl-4-pentene-1,2-diol (51b).
  According to AAV 10, from 51b-Ac ₄ , 3.82 g (quant. Yield).
• 105. (2RS) -1-O-α-D-galactopyranosyl-4-pentene-1,2-diol (52a).
  According to AAV 6, AAV 7, AAV 9 and AAV 10, from 2a, 10.0 g (82% for 4 steps).
  The product corresponds to that produced according to AAV 8 from 2a-O.
• 106. (2RS) -1-O-β-D-galactopyranosyl-4-pentene-1,2-diol (52b).
  According to AAV 6, AAV 7, AAV 9 and AAV 10, from 2b, 6.9 g (61% for 4 steps).
• 107. (2RS) -1-O- (2-acetamido-2-deoxy-α-D-glucopyranosyl) -4-pentene-1,2-diol (53a).
  According to AAV 6, AAV 7, AAV 9 and AAV 10, from 3a, 7.5 g (83% for 4 steps).
  The product corresponds to that produced according to AAV 8 from 3a.
• 108. (2RS) -1-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl) -4-pentene-1,2-diol (53b).
  According to AAV 7, AAV 9 and AAV 10, from 3b-Ac ₃ , 130 g (68% for 3 steps).
  The product corresponds to that produced according to AAV 8 from 3b.
• 109. (2RS) -1-O- (Methyl-β-D-glucopyranuronosyl) -4-pentene-1,2-diol (55b).
  According to AAV 10, from 55b-Ac ₃ , 2.37 g (quant. Yield).
• 110. (2RS) -1-O- (methyl-β-D-galactopyranuronosyl) -4-pentene-1,2-diol (56b).
  According to AAV 10, from 56b-Ac ₃ , 2.64 g (79%).
• 111. (2RS) -1-O-β-D-xylopyranosyl-4-pentene-1,2-diol (59b).
  According to AAV 10, from 59b-Ac ₃ , 5.06 g (quant. Yield).
• 112. (2RS) -1-O-β-D-ribopyranosyl-4-pentene-1,2-diol (61b).
  According to AAV 6, AAV 7, AAV 9 and AAV 10, from 11b, 7.34 g (60%).
• 113. (2RS) -1-O- (4-O-β-D-glucopyranosyl-β-D-glucopyranosyl) -4-pentene-1,2-diol (62b).
  According to AAV 10, from 62b-Ac ₇ , 4.26 g (quant. Yield).
• 114. (2RS) -1-O- (4-O-β-D-galactopyranosyl-β-D-glucopyranosyl) -4-pentene-1,2-diol (63b).
  According to AAV 10, from 63b-Ac ₇ , 550 mg (99%).
• 115. (2RS) -1-O-β-D-glucopyranosyl-2-methyl-4-pentene-1,2-diol (67b).
  According to AAV 10, from 67b-Ac ₄ , 2.78 g (quant. Yield).
• 116. (2RS) -1-O-β-D-galactopyranosyl-2-methyl-4-pentene-1,2-diol (68b).
  According to AAV 10, from 68b-Ac ₄ , 2.78 g (quant. Yield).
• 117. (2RS) -1-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl) -2-methyl-4-pentene-1,2-diol (69b).
  According to AAV 10, from 69b-Ac ₃ , 536 mg (quant. Yield).
• 118. (2RS) -2- (β-D-glucopyranosyloxymethyl) -2-hydroxy-4-pentenoic acid methyl ester (75b).
  According to AAV 10, from 75b-Ac ₄ , 1.58 g (98%).
• 119. (2RS) -2-O-β-D-glucopyranosyl-3-buten-1,2-diol (26b).
  According to AAV 10, from 26b-Ac ₄ Tr, 430 mg (86%).
• 120. (2RS) -2-O- (methyl-β-D-glucopyranuronosyl) -3-buten-1,2-diol (28b).
  According to AAV 10, from 28b-Ac ₃ Tr, 170 mg (79%).
• 121. (2RS) -2-O- (4-O-α-D-glucopyranosyl-β-D-glucopyranosyl) -3-buten-1,2-diol (29b).
  According to AAV 10, from 29b-Ac ₇ Tr, 1.33 g (97%).
• 122. 1,1-bis (β-D-glucopyranosyloxymethyl) -3-huten-1-ol (87b).
  According to AAV 10, from 87b-Ac ₈ , 4.56 g (quant. Yield).

Work regulations for derivatization

• 123. (2RS) -1-O- (4,6-O-benzylidene-α-D-galactopyranosyl) -4-pentene-1,2-diol (118).
  A 1.6 M suspension of 52a in benzaldehyde is mixed with 1.5 equivalents of benzaldehyde dimethyl acetal and a catalytic amount of p-toluenesulfonic acid and stirred for 60 min at room temperature until a homogeneous solution is obtained and TLC control signals complete conversion. It is neutralized with a few drops of triethylamine, diluted with MeOH / H ₂ O (2: 1) and excess reagent is removed by extraction with pentane. The methanolic phase is i. Vac. concentrated and the crude product chromatographed on silica gel. Yield 3.35 g (60%).
• 124. (2RS) -1-O- (4,6-O-furfurylidene-α-D-galactopyranosyl) -4-pentene-1,2-diol (119).
  A 1.6 M suspension of 52a in furfural is mixed with 1.5 eq of 2-furaldehyde diethylacetal and a catalytic amount of p-toluenesulfonic acid and stirred for 40 min at room temperature until a homogeneous solution is obtained and DC control signals complete conversion. It is neutralized with a few drops of triethylamine, diluted with MeOH / H ₂ O (2: 1) and excess reagent removed by extraction with pentane. The methanolic phase is i. Vac. concentrated and the crude product chromatographed on silica gel. Yield 5.18 g (79%).
  13 C-NMR (D ₂ O, 75 MHz): d = 151.99 (Ar), 146.26 (Ar), 137.30 / 137.06 (C-4), 120.64 / 120.47 (C-5), 113.32 (Ar), 111.56 ( Ar), 102.19 / 101.70 (C-1 '), 97.95 (ArCH), 79.09 (C-4'), 74.39 / 73.90 (C-1), 72.46 / 72.08 (C-2), 71.65 (C-6 ' ), 70.94 / 70.89 (C-3 ') *, 70.61 / 70.59 (C-2') *, 65.64 / 65.62 (C-5 '), 39.90 (C-3).
• 125. Methyl-5-O-allyl-2,3-O-isopropylidene-β-D-ribofuranoside (82b).
  1.3 g of sodium hydride is suspended in 5 ml of dry THF and a 1.2 M solution of b-methyl-2,3-O-isopropylidene-D-ribofuranoside (10.0 g, 49 mmol; from ribose, methanol and acetone according to AAV 1) in THF slowly added dropwise with stirring. After 2 h, a 4 M solution of allyl bromide (5.5 ml, 64 mmol) in THF is added dropwise and, after stirring for a further 14 h, hydrolyzed with ice. The solution is i. Vac. concentrated to 1/3 of the volume and then extracted three times with ether. After drying the organic phase over MgSO ₄ and concentrating the solution i. Vac. 11.5 g (96%) 82b are obtained as a colorless liquid.

General working instructions for ozonolysis (AAV 11)

Ozone is introduced into a 10-200 mM solution of the saccharide in methanol (approx. 1% water content) at -78 ° C until a blue color remains. Excess ozone is expelled by introducing air. Dimethyl sulfide is added dropwise with stirring (0.5 ml per 1 mmol olefin). The solution is slowly thawed and left to stand at room temperature until a test with AI starch paper is negative (4-16 h), then concentrated in vacuo. The remaining syrup is dissolved several times in methanol (5-10% water content) and again concentrated in vacuo. The carbonyl compounds contain varying proportions of dimethyl sulfoxide and are used for enzymatic reactions without purification.

• 126. (3RS) -4-O-α-D-glucopyranosyl-3,4-dihydroxybutanal (51a-0).
  According to AAV 11, from 51a (2.64 g); according to NMR mixture of free aldehyde, hydrate and diastereomeric methyl hemiacetals.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 105.62 (C-1), 105.46 (C-1), 101.43 (C-1 '), 100.97 (C-1'), 75.85 (C-3 '), 74.71 (C-2'), 74.48 (C-4), 74.29 (C-5 '), 74.16 (C-4), 72.38 (C- 4'), 69.70 (C-3), 69.32 ( C-3), 63.32 (C-6 '), 56.28 (CH ₃ O), 38.73 (C-2).
• 127. (3RS) -4-O-β-D-glucopyranosyl-3,4-dihydroxybutanal (51b-O).
  According to AAV II, from 51b (3.82 g); according to NMR mixture of free aldehyde, hydrate and diastereomeric methyl hemiacetals.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 105.56 (C-1 '), 105.18 (C-1'), 91.32 (C-1), 78.70 (C-3 '), 78.42 (C- 5 '), 76.69 (C-3), 76.37 (C-3 *), 75.90 (C-2'), 72.43 (C-4 '), 69.72 (C- 4), 63.52 (C-6'), 51.69 (CH ₃ O), 44.40 (C-2).
• 128. (3RS) -4-O-α-D-galactopyranosyl-3,4-dihydroxybutanal (52a-O).
  According to AAV 11, from 52a (750 mg).
  ¹³ C NMR (D ₂ O, 75 MHz): d = 101.37 / 100.91 (C-1 '), 91.22 (C-1), 74.33 / 73.98 (C-4), 73.67 (C-5'), 72.08 (C-3 '), 71.93 (C-4'), 71.07 (C-2 '), 69.93 / 69.58 (C-3), 63.88 (C-6'), 43.04 (C-2).
• 129. (3RS) -4-O- (4-O-β-D-glucopyranosyl-β-D-glucopyranosyl) -3,4-dihydroxybutanal (62b-O).
  According to AAV 11, from 62b (3.84 g); according to NMR mixture of free aldehyde and hydrate.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 105.40 (C-1 °), 81.47, 78.79, 78.30, 77.55, 77.01, 75.96, 72.26, 63.39 (CH ₂ ), 62.83 (CH ₂ ).
• 130. (3RS) -4-O-β-D-glucopyranosyl-3,4-dihydroxy-3-methylbutanal (67b-O).
  According to AAV 11, from 67b (2.50 g); according to NMR mixture of free aldehyde (approx. 20%), aldehyde hydrate and diastereomeric methyl hemiacetals.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 105.8 (C-1 '), 105.66 (C-1'), 105.6 (C-1 '), 105.4 (C-1'), 91 (C -1), 79.31 (C-4), 78.72, 78.42, 76.02, 75.9, 74.32 (C-3), 74.07 (C-3), 73.9 (C-3), 72.52, 72.5, 63.53 (C-6 ' ), 54.0 (CH ₃ O), 41 (C-2), 26.28 (3-CH ₃ ), 25.92 (3-CH ₃ ).
• 131. (3RS) -4-O-β-D-galactopyranosyl-3,4-dihydroxy-3-methylbutanal (68b-O).
  According to AAV 11, from 68b (2.50 g); according to NMR mixture of free aldehyde (approx. 20%), aldehyde hydrate and diastereomeric methyl hemiacetals.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 209.56 (C-1), 106.30 (C-1 '), 105.81 (C-1'), 102.50 (C-1 '), 91.24 (C- 1), 79.34 (C-4), 79.12 (C-4), 78.17, 77.94, 76.54, 75.46, 74.16 (C- 3), 73.9 (C-3), 73.74, 73.31, 71.48, 71.14, 69.38, 63.81 (C-6 '), 63.60 (C-6'), 54.06 (CH ₃ O), 53.99 (CH ₃ O), 47.83 (C-2), 45.70 (C-2), 45.18 (C-2), 26.28 (3-CH ₃ ), 25.78 (3-CH ₃ ).
• 132. (2RS) -2- (β-D-glucopyranosyloxymethyl) -2-hydroxy-4-oxobutanoic acid methyl ester (75b-O).
  According to AAV 11, from 75b (1.58 g); according to NMR mixture of free aldehyde, aldehyde hydrate and diastereomeric methyl hemiacetals.
  ¹³ C NMR (75.4 MHz; D ₂ O): d = 177.81 (C-1), 177.4 (C-1), 105.73 (C-1 '), 105.37 (C-1'), 104.58 (C-1 '), 89.79 (C-4), 78.61, 78.3, 78.21, 77.79, 75.80, 75.53, 72.28, 65.57 (CH ₂ (Et-Ester)), 63.35 (C-6'), 57.82 (CH ₃ ), 56.47 (CH ₃ ), 55.66 (CH ₃ ), 51.58 (CH ₃ ), 44.86 (C-3), 41.01 (C-3), 15.97 (CH ₃ (Et ester)).
• 133. 3,3-bis (β-D-glucopyranosyloxymethyl) -3-hydroxypropanal (87b-O).
  According to AAV 11, from 87b (4.11 g); according to NMR mixture of free aldehyde (40%), aldehyde hydrate and diastereomeric methyl hemiacetals.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 208.55 (C-1), 105.65 (C-1 '), 105.54 (C-1'), 90.75 (C-1), 78.72 (C-3 '), 78.40 (C-5'), 76.20 (C-3), 75.96 (C-2 '), 75.50 (3-CH ₂ OR), 75.20 (3-CH ₂ OR), 72.51 (C-4' ), 72.46 (C-4 '), 63.53 (C-6'), 51.72 (CH ₃ O), 50.16 (CH ₃ O), 44 (C-2).

General working procedure for enzymatic aldol addition (AAV 12)

The aldehyde dissolved in a little water is taken up in an aqueous solution of DHAP (80-100 mM), adjusted to pH 6.8-7.3 and aldolase (FruA, RhuA or FucA; 50-150 U per 1 mmol aldehyde) is added. If the conversion is incomplete, DHAP and possibly enzyme are added after 1 or 2 days, and the pH is checked. After the reaction has ended (TLC control, 1-7 d), the ketosephosphate is bound to anion exchanger (HCO ₃ ⁻ form) and eluted with 150-200 mM triethyl ammonium bicarbonate buffer. The buffer is i. Vac., Repeated absorption in water and subsequent concentration removed. The ketose phosphate is obtained as the sodium salt by cation exchange (sodium-loaded cation exchanger or acidic ion exchanger and neutralization with sodium hydroxide solution).

• 134. 5-O-β-D-glucopyranosyl-D-sorbose-1-phosphate (107-P).
  According to AAV 11 and AAV 12 (RhuA); from 26b; 368 mg (50% D-enantiomer) disodium salt.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 103.57 (C-1 '), 99.19 (d, C-2; J = 9.1), 78.98, 78.41, 78.04, 75.37, 74.30, 72.75, 72.09, 69.32, 63.24 (C-6), 62.21 (C-6).

General working procedure for enzymatic dephosphorylation (AAV 13)

A 20-100 mM solution of the isolated or crude ketose phosphate is adjusted to pH 7.5 (6.0) and mixed with 80-200 (20) U / mmol alkaline (acidic) phosphatase. After conversion is complete (1-4 d), the mixture is filtered through activated carbon and the solution (optionally) is desalted by filtration through an ion exchanger (HCO ₃ ⁻ and H⁺ form). The solution is i. Vac. concentrated and the remaining syrup on silica gel, on Ca ²⁺ loaded cation exchanger, on Biogel P2 or on an activated carbon / diatomaceous earth (1/1) mixture.

• 135. 7-O-α-D-galactopyranosyl-5-deoxy-D-arabino-hept-2-ulopyranose (101a) and 7-O-α-D-galactopyranosyl-5-deoxy-L-xylo-hept-2 -ulopyranose (102a).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (acid phosphatase); from 52a; 118 mg (12%) 101a and 175 mg (18%) 102a.
  (101a): ¹³ C-NMR (D ₂ O, 125 MHz): d = 101.05 (C-2), 100.85 (C-1 '), 73.80 (C-5'), 72.30 (C-7), 72.26 (C-3 '), 72.07 (C-4'), 71.20 (C-2 '), 70.75 (C-4), 68.06 (C-3), 67.17 (C-1), 66.74 (C-6) , 64.02 (C-6 '), 30.61 (C-5).
  (102a): ¹³ C NMR (D ₂ O, 125 MHz): d = 101.64 (C-1 '), 100.85 (C-2), 74.87 (C-3), 73.79 (C-5'), 72.83 (C-7), 72.16 (C-3 '), 72.02 (C-4'), 71.10 (C-2 '), 70.92 (C-4 *), 70.78 (C-6 *), 66.58 (C- 1), 64.02 (C-6 '), 37.40 (C-5).
• 136. 5-Deoxy-7-O-β-D-xylopyranosyl-D-arabino-hept-2-ulopyranose (101b) and 5-deoxy-7-O-β-D-xylopyranosyl-L-xylo-hept-2 -ulopyranose (102b).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 59b; 130 mg (70%) mixture of 101b and 102b in a ratio of 3: 4.
  ¹³ C NMR (75.4 MHz; D ₂ O): d = 106.05 (C-1 '), 105.50 (C-1'), 100.66 (C-2), 100.58 (C-2), 78.46, 78.30, 78.21 , 75.61, 75.53, 74.97 (C-7), 74.66, 74.22 (C-7), 71.83, 70.65, 70.36, 70.05, 68.46, 68.10, 67.79 (C-5 '), 67.51 (C-5'), 65.52 (C-1), 63.89 (C-1), 37.06 (C-5), 30.43 (C-5).
• 137. 7-O- (2-Acetamido-2-deoxy-α-D-glucopyranosyl) -5-deoxy-D-arabino hept-2-ulopyranose (101c) and 7-O- (2-acetamido-2-deoxy -α-D-glucopyrano syl) -5-deoxy-L-xylo-hept-2-ulopyranose (102c).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 53a; 39 mg (1.4%) 101c and 23 mg (0.8%) 102c.
  (101c): C-NMR (D ₂ O, 75 MHz): d = 177.31 (Ac), 101.09 (C-2), 99.63 (C-1 '), 74.79 (C-5'), 73.91 (C- 3 '), 72.78 (C-4'), 72.22 (C-7), 70.74 (C-4), 68.02 (C-3), 67.17 (C-1), 66.82 (C-6), 63.37 (C -6 '), 56.38 (C-2'), 30.69 (C-5), 24.79 (Ac).
  (102c): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.28 (Ac), 100.95 (C-1 '), 100.50 (C-2), 74.90 (C-3), 74.84 (C- 5 '), 73.78 (C-3'), 73.01 (C-7), 72.84 (C-4 '), 70.95 (C-4), 70.86 (C-6), 66.59 (C-1), 63.45 ( C-6 '), 56.50 (C-2'), 37.43 (C-5), 24.75 (Ac).
• 138.7-O- (2-acetamido-2-deoxy-β-D-gluc 11832 00070 552 001000280000000200012000285911172100040 0002019709787 00004 11713opyranosyl) -5-deoxy-D-arabino hept-2-ulopyranose (101d) and 7-O- (101d 2-Acetamido-2-deoxy-β-D-glucopyrano syl) -5-deoxy-L-xylo-hept-2-ulopyranose (102d).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 53b; 330 mg (11%) 101d and 690 mg (23%) 102d.
  (101d): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.48 (Ac), 104.33 (C-1 '), 100.73 (C-2), 78.66 (C-5'), 76.52 (C -3 '), 75.15 (C-7), 72.68 (C-4'), 70.59 (C-4), 68.34 (C-3), 67.23 (C-1), 66.89 (C-6), 63.51 ( C-6 '), 58.31 (C-2'), 30.53 (C-5), 25.02 (Ac).
  (102d): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.32 (Ac), 104.10 (C-1 '), 100.72 (C-2), 78.62 (C-5'), 76.52 (C -3 '), 74.89 (C-3), 74.33 (C-7), 72.66 (C-4'), 70.75 (C-4), 70.32 (C-6), 66.59 (C-1), 63.49 ( C-6 '), 58.23 (C-2'), 37.31 (C-5), 25.07 (Ac).
• 139. 7-O-α-D-galactopyranosyl-5-deoxy-D-ribo-hept-2-ulopyranose (103a) and 7-O-α-D-galactopyranosyl-5-deoxy-L-lyxo-hept-2 -ulopyranose (104a).
  According to AAV 11, AAV 12 (FucA) and AAV 13 (alkaline phosphatase); from 52a; 66 mg (8%) 103a and 19 mg (2%) 104a.
  (103a): ¹³ C-NMR (D ₂ O, 75 MHz): d = 101.55 (C-2), 100.83 (C-1 '), 73.83 (C-5'), 72.26 (C-3), 72.08 (C-4 '), 72.02 (C-7), 71.15 (C-2'), 71.04 (C-4), 68.78 (C-3), 66.60 (C-6), 66.44 (C-1), 64.06 (C-6 '), 35.79 (C-5).
  (104a): ¹³ C-NMR (D ₂ O, 75 MHz): d = 101.84 (C-1 '), 101.35 (C-2), 73.89 (C-5'), 73.27 (C-7), 72.24 (C-3 '), 72.12 (C-4') '71.49 (C-4), 71.23 (C-2'), 70.63 (C-3), 68.26 (C-6), 67.28 (C-1) , 64.12 (C-6 '), 31.91 (C-5).
• 140. 5-Deoxy-7-O-β-D-xylopyranosyl-D-ribo-hept-2-ulopyranose (103b) and 5-deoxy-7-O-β-D-xylopyranosyl-L-lyxo-hept-2 -ulopyranose (104b).
  According to AAV 11, AAV 12 (FucA) and AAV 13 (alkaline phosphatase); from 59b; 20 mg (3%) mixture of 103b and 104b in a ratio of 1: 2.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 106.0 (C-1 '), 105.49 (C-1'), 101.2 (C-2), 100.92 (C-2), 78.25, 75.5 (C -7), 71.80, 67.74 (C-5 '), 67.4 (C-5'), 67 (C-1), 63 (C-1), 35.51 (C-5), 31.49 (C-5).
• 141. 7-O- (2-acetamido-2-deoxy-α-D-glucopyranosyl) -5-deoxy-D-ribo-hept-2-ulopyranose (103c) and 7-O- (2-acetamido-2- deoxy-α-D-glucopyranosyl) -5- deoxy-L-lyxo-hept-2-ulopyranose (104c).
  According to AAV 11, AAV 12 (FucA) and AAV 13 (alkaline phosphatase); from 53a; 104 mg (4%) 103c and 26 mg (1%) 104c.
  (103c): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.26 (Ac), 101.56 (C-2), 99.65 (C-1 '), 74.80 (C-5'), 73.84 (C -3 '), 72.82 (C-4'), 71.95 (C-7), 71.04 (C-4), 68.96 (C-3), 66.78 (C-6), 66.53 (C-1), 63.41 ( C-6 '), 56.44 (C-2'), 35.81 (C-5), 24.77 (Ac).
  (104c): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.30 (Ac), 101.40 (C-2), 100.58 (C-1 '), 74.86 (C-5'), 73.81 (C -3 '), 73.25 (C-7), 72.86 (C-4'), 71.41 (C-4), 70.54 (C-3), 68.27 (C-6), 67.19 (C-1), 63.46 ( C-6 '), 56.47 (C-2'), 31.85 (C-5), 24.77 (Ac).
• 142. 7-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl) -5-deoxy-D-ribo-hept-2-ulopyranose (103d) and 7-O- (2-acetamido-2- deoxy-β-D-glucopyranosyl) -5-deoxy-L-lyxo-hept-2-ulopyranose (104d).
  According to AAV 11, AAV 12 (FucA) and AAV 13 (alkaline phosphatase); from 53b; 372 mg (12%) 103d and 62 mg (2%) 104d.
  (103d): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.39 (Ac), 104.45 (C-1 '), 101.48 (C-2), 78.67 (C-5'), 76.51 (C -3 '), 74.84 (C-7), 72.70 (C-4'), 70.98 (C-4), 68.72 (C-3), 66.63 (C-6), 66.56 (C-1), 63.54 ( C-6 '), 58.31 (C-2'), 35.45 (C-5), 24.98 (Ac).
  (104d): ¹³ C-NMR (D ₂ O, 125 MHz): d = 177.56 (Ac), 104.00 (C-1 '), 101.18 (C-2), 78.74 (C-5'), 76.64 (C -3 '), 74.83 (C-7), 72.74 (C-4'), 70.74 (C-4) *, 70.72 (C-3) *, 68.24 (C-6), 67.35 (C-1), 63.57 (C-6 '), 58.31 (C-2'), 31.98 (C-5), 25.06 (Ac).
• 143. 7-O-α-D-galactopyranosyl-5-deoxy-D-xylo-hept-2-ulopyranose (105a) and 7-O-α-D-galactopyranosyl-5-deoxy-L-arabino-hept-2 -ulopyranose (106a).
  According to AAV 11, AAV 12 (RhuA) and AAV 13 (alkaline phosphatase); from 52a; 550 mg (19%) 105a and 120 mg (4%) 106a.
  (105a): ¹³ C-NMR (D ₂ O, 75 MHz): d = 101.24 (C-2), 100.85 (C-1 '), 74.76 (C-3), 73.79 (C-5'), 72.21 (C-3 '), 72.04 (C-4'), 71.91 (C-7), 71.10 (C-2 '), 70.97 (C-4), 70.34 (C-6), 66.54 (C-1) , 64.03 (C-6 '), 37.25 (C-5).
  (106a): ¹³ C-NMR (D ₂ O, 75 MHz): d = 101.80 (C-1 '), 100.80 (C-2), 73.83 (C-5'), 73.35 (C-7), 72.24 (C-3 '), 72.10 (C-4'), 71.27 (C-2 '), 70.56 (C-4), 68.33 (C-3), 67.33 (C-6), 67.25 (C-1) , 64.05 (C-6 '), 30.90 (C-5).
• 144. 5-deoxy-7-O-β-D-xylopyranosyl-D-xylo-hept-2-ulopyranose (105b) and 5-deoxy-7-O-β-D-xylopyranosyl-L-arabino-hept-2 -ulopyranose (106b).
  According to AAV 11, AAV 12 (RhuA) and AAV 13 (alkaline phosphatase); from 59b; 66 mg (40%) mixture of 105b and 106b in a ratio of 3: 8.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 106.10 (C-1 '), 105.59 (C-1'), 101.11 (C-2), 100.77 (C-2), 78.56, 78.36, 75.68 , 75.63, 74.63 (C-7), 74.52 (C-7), 71.89, 70.62, 70.54, 68.54, 68.16, 67.88 (C-5 '), 67.20 (C-1), 65.59 (C-1), 37.04 (C-5), 31.56 (C-5).
• 145. 7-O- (2-acetamido-2-deoxy-α-D-glucopyranosyl) -5-deoxy-D-xylo-hept-2-ulopyranose (105c) and 7-O- (2-acetamido-2- deoxy-α-D-glucopyranosyl) -5- deoxy-L-arabino-hept-2-ulopyranose (106c).
  According to AAV 11, AAV 12 (RhuA) and AAV 13 (alkaline phosphatase); from 53a; 150 mg (6%) 105c and 201 mg (7%) 106c.
  (105c): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.30, 101.21, 100.40.74.81.74.76.73.80, 72.79, 71.94, 70.97, 70.38, 66.49, 63.41, 56.48, 37.30, 24.76.
  (106c): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.30, 101.46, 100.98, 74.83, 73.90, 73.35, 72.81, 70.60, 68.25, 67.29, 67.19, 63.39, 56.49, 30.83, 24.79.
• 146. 7-O- (2-acetamido-2-deoxy-β-D-glucopyranosyl) -5-deoxy-D-xylo-hept- 2-ulopyranose (105d) and 7-O- (2-acetamido-2- deoxy-β-D-glucopyranosyl) -5-deoxy-L-arabino-hept-2-ulopyranose (106d).
  According to AAV 11, AAV 12 (RhuA) and AAV 13 (alkaline phosphatase); from 53b; 247 mg (8%) 105d and 440 mg (14%) 106d.
  (105d): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.45 (Ac), 104.47 (C-1 '), 100.83 (C-2), 78.63 (C-5'), 76.56 (C -3 '), 74.76 (C-7), 74.67 (C-3), 72.65 (C-4'), 70.87 (C-4), 70.36 (C-6), 66.62 (C-1), 63.48 ( C-6 '), 58.23 (C-2'), 36.97 (C-5), 25.02 (Ac).
  (106d): ¹³ C-NMR (D ₂ O, 75 MHz): d = 177.52 (Ac), 103.82 (C-1 '), 100.64 (C-2), 78.68 (C-5'), 76.62 (C -3 '), 74.76 (C-7), 72.71 (C-4'), 70.55 (C-4), 68.41 (C-3), 67.31 (C-1), 66.55 (C-6), 63.53 ( C-6 '), 58.29 (C-2'), 30.86 (C-5), 25.08 (Ac).
• 147. 7-O- (4,6-O-benzylidene-α-D-galactopyranosyl) -5-deoxy-L-xylo-hept-2-ulopyranose (120).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 118; 664 mg (37%) 120.
  ¹³ C-NMR (D ₂ O, 75 MHz): d = 139.76 (Ph), 132.47 (Ph), 131.37 (Ph), 129.04 (Ph), 103.65 (PhCH), 102.15 (C-1 '), 100.73 ( C-2), 78.82 (C-4 '), 74.84 (C-3), 72.93 (C-7), 71.59 (C-6'), 70.90 (C-3 ') *, 70.87 (C-4) , 70.66 (C-6), 70.60 (C-2 ') *, 66.57 (C-1), 65.67 (C-5'), 37.24 (C-5).
• 148. 7-O- (2,3-O-isopropylidene-β-D-ribofuranosyl) -5-deoxy-D-ribo-hept-2-ulopyranose (121) and 7-O- (2,3-O- Isopropylidene-β-D-ribofuranosyl) -5-deoxy-L-lyxo-hept-2-ulopyranose (122).
  According to AAV 11, AAV 12 (FucA) and AAV 13 (alkaline phosphatase); from 66b; 400 mg (22%) 121 and 270 mg (15%) 122.
  (121): ¹³ C-NMR (D ₂ O, 75 MHz): d = 115.75 (C _q ), 110.94 (C-1 '), 101.34 (C-2), 89.87 (C-4'), 87.20 ( C-3 ') *, 83.78 (C-2') *, 72.86 (C-7), 70.85 (C-4), 68.76 (C-3), 66.69 (C-6), 66.47 (C-1) , 65.31 (C-5 '), 35.72 (C-5), 28.06 (CH ₃ ), 26.42 (CH ₃ ).
  (122): ¹³ C-NMR (D ₂ O, 75 MHz): d = 115.68 (C _q ), 110.04 (C-1 '), 101.14 (C-2), 89.84 (C-4'), 87.19 ( C-3 ') *, 83.74 (C-2') *, 72.30 (C-7), 70.42 (C-4) **, 70.25 (C- 3) **, 68.17 (C-6), 67.09 ( C-1), 65.30 (C-5 '), 31.59 (C-5), 28.04 (CH ₃ ), 26.40 (CH ₃ ).
• 149. 5-O-β-D-glucopyranosyl-D-sorbose (107).
  According to AAV 11, AAV 12 (RhuA) and AAV 13 (alkaline phosphatase); from 26b; 28 mg (47% based on D enantiomer).
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 103.87 (C-1 '), 100.36 (C-2), 79.47, 78.74, 78.35, 75.68, 74.82, 73.17, 72.42, 66.24 (C-1) , 63.56, 62.40.
• 150. 5-O-β-D-glucopyranosyl-D-fructose (108) and 5-O-β-D-glucopyranosyl-L-sorbose (109).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 26b; 154 mg (45%) 108 and 109 in a 6: 5 ratio.
  (108): ¹³ C-NMR (75.4 MHz; D ₂ O): d = 103.70 (C-1 '), 100.98 (C-2), 79.61 (C-5), 78.67 (C-5'), 78.31 (C-3 '), 75.59 (C-2'), 75.46 (C-4), 72.42 (C-4 '), 71.53 (C-3), 66.34 (C-1), 63.82 (C-6) , 63.50 (C-6 ').
  (109): ¹³ C-NMR (75.4 MHz; D ₂ O): d = 106.16 (C-1 '), 100.23 (C-2), 81.85 (C-5), 78.67 (C-5'), 78.38 (C-3 '), 76.14 (C-2'), 72.96 (C-3), 72.36 (C-4 '), 70.54 (C-4), 66.23 (C-1), 63.78 (C-6) , 63.41 (C-6 ').
• 151. 6-O-β-D-glucopyranosyl-D-fructose (110).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 30b; 140 mg (38%).
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 105.43, 104.60, 100.87, 98.69, 82.03, 78.72, 78.54, 78.36, 77.90, 77.46, 76.93, 75.95, 75.56, 74.23, 73.82, 72.42, 71.99, 70.35 , 66.68, 66.17, 65.44, 63.54.
• 152. 5-O-β-D-galactopyranosyl-D-sorbose (111) and 5-O-β-D-galactopyranosyl-L-fructose (112).
  According to AAV 11, AAV 12 (RhuA) and AAV 13 (alkaline phosphatase); from 27b; 1.54 g (90%) 111 and 112 in a 1: 1 ratio.
  (111): ¹³ C-NMR (75.4 MHz; D ₂ O): d = 104.50 (C-1 '), 100.33 (C-2), 79.51, 78.09, 75.41, 74.92, 73.44, 73.06, 71.44, 66.21 ( C-1), 63.96 (C-6), 62.41 (C-6). (112): ¹³ C-NMR (75.4 MHz; D ₂ O): d = 107.52 (C-1 '), 100.97 (C-2), 82.45, 78.04, 75.39, 73.97, 72.60, 71.49, 70.61, 66.60 ( C-1), 65.50 (C-6), 63.82 (C-6).
• 153. 5-deoxy-7-O-β-D-galactopyranosyl-6-C-methyl-D-xylo-hept-2-ulo pyranose (113) and 5-deoxy-7-O-β-D-galactopyranosyl- 6-C-methyl-L-arabino hept-2-ulopyranose (114).
  According to AAV 11, AAV 12 (RhuA) and AAV 13 (alkaline phosphatase); from 68b; 130 mg (18%) mixture of 113 and 114 in a ratio of 1: 1.
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 106.57 (C-1 '), 106.25 (C-1'), 102.01 (C-2), 101.89 (C-2), 80.27 (C-6 ), 79.42 (C-6), 78.86 (C-7), 78.76 (C-7), 77.95, 77.57, 75.53, 75.42, 74.76, 74.70, 73.73, 73.64, 73.56, 71.44, 68.22, 68.13, 67.12 (C -2), 66.82 (C- 2), 63.78 (C-6 '), 42.53 (C-5), 41.17 (C-5), 28.99 (6-CH ₃ ), 25.84 (6-CH ₃ ).
• 154. 5-Deoxy-7-O- (4-O-β-D-glucopyranosyl-β-D-glucopyranosyl) -L-xylohept-2-ulopyranose (115).
  According to AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 62b-O; 55 mg (22%).
  ¹³ C-NMR (75.4 MHz; D ₂ O): d = 105.7 (C-1 '), 101.0 (C-2), 101.1, 37.3 (C-5).
• 155. 7-O-β-D-ribopyranosyl-5-deoxy-D-arabino-hept-2-ulopyranose (116) and 7-O-β-D-ribopyranosyl-5-deoxy-L-xylo-hept-2 -ulopyranose (117).
  According to AAV 11, AAV 12 (FruA) and AAV 13 (alkaline phosphatase); from 61b; 218 mg (11%) 116 and 441 mg (23%) 117.
  (116): ¹³ C NMR (D ₂ O, 75 MHz): d = 103.62 (C-1 '), 100.71 (C-2), 74.21 (C-7), 72.88 (C-2'), 70.62 (C-4) *, 70.53 (C-4 ') *, 70.38 (C-3') *, 68.30 (C-3), 67.24 (C-1), 67.15 (C-6), 66.01 (C- 5 '), 30.70 (C-5).
  (117): ¹³ C-NMR (D ₂ O, 75 MHz): d = 102.87 (C-1 '), 100.81 (C-2), 74.75 (C-3), 73.14 (C-7), 72.76 ( C-2 '), 70.85 (C-4), 70.46 (C-6) *, 70.29 (C-4') *, 70.14 (C-3 ') *, 66.58 (C-1), 65.99 (C- 5 '), 37.16 (C-5).

Claims (4)

1. Compounds of the general formula (I)
as well as pharmaceutically acceptable salts thereof,
in which
A represents CH ₂ or a radical of the formula
stands,
B represents CH ₂ , a bond or a radical of the formula
stands,
where A and B should not simultaneously represent a CH ₂ group,
R represents hydrogen or phosphate,
in which
R ^{1 represents} an optionally substituted glycosyl radical,
R ² and R ^{3 may be the} same or different and represent hydrogen, hydroxy, hydroxyalkyl, glycosyloxy, glycosyloxyalkyl, alkoxyalkyl, dialkoxy alkyl or azido or
R ² and R ³ together with the carbon atom to which they are attached represent a 5- or 6-membered saturated ring which can contain up to 2 oxygen atoms, where the ring can be substituted up to twice by lower alkyl, and
R ^{4 represents} hydrogen, alkyl, alkyloxycarbonyl, carboxy or glycosyloxy alkyl,
X represents -O-, -OCH ₂ -, -N (alkanoyl) -, -S-, -SCH ₂ - or a bond,
R ¹ ', independently of R ¹ , can have the meanings given for R ¹ ,
R ³ 'independently of R ³ can have the meanings given for R ³ and
X 'can have the meanings given for X independently of X. 2. Compounds of general formula (I) according to claim 1, characterized in that
A, B, R, R ¹ and R ^{1 '} , which have the meaning given in claim 1 and
R ² and R ^{3 may be the} same or different and stand for hydrogen, hydroxy, hydroxymethyl, glycosyloxy, glycosyloxymethyl, alkoxyalkyl, dialk oxyalkyl or azido or together represent -OC (CH ₃ ) ₂ OCH ₂ and
R ^{4 represents} hydrogen, methyl, methoxycarbonyl, carboxy or glycosyl oxymethyl,
X represents -O-, -OCH ₂ -, -N (acetyl) -, -S-, -SCH ₂ - or a bond,
R ³ 'independently of R ³ can have the meanings given for R ³ and
X 'can have the meanings given for X independently of X. 3. Process for the preparation of enantiomerically pure and diastereomerically pure oligosaccharides and their derivatives, characterized in that

• (1) produces a glycoside according to standard processes, which contains a polar addition-capable double bond in the aglycon part,
• (2) a C-nucleophilic reagent, which itself contains an aldehyde function in masked form, is added to the polar double bond of the glycoside with CC linkage, a heteronucleophile being produced from the original double bond,
• (3) the aldehyde function is released from the masked aldehyde function of the compounds obtained in step (2) by a corresponding chemical reaction and enzyme-catalyzed to the aldehyde obtained in the sense of an aldol reaction by means of a specific aldolase dihydroxyacetone phosphate (DHAP) with CC linkage, where a typical sugar ring structure of formula (I) is formed by cyclization via the previously generated heteronucleophile to the ketone group of the DHAP unit.

4. The method according to claim 3, characterized in that compounds of the general formula (I) according to claim 1 are prepared by

• (1) prepares compounds of the formula (IV) by known processes,
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given in claim 1 and
  R ⁵ and R ^{6 can be the} same or different and represent hydrogen, alkyl or aryl,
  and by converting the double bond into compounds of the formula (V),
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given in Claim 1,
• (2) receives compounds of the formula (VI) by reacting compounds of the formula (V) with organometallic reagents which have a double bond,
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given in claim 1 and in which
  Y represents -CH ₂ -, CH (CH ₃ ) -, -C (CH ₃ ) ₂ - or a bond and
  Z for = CH ₂ or = C = CH ₂
  stands,
• (3) releases the aldehyde of the formula (VII) from the masked aldehyde function of the compounds of the formula (VI) by a corresponding chemical reaction,
  in the
  R ¹ , R ² , R ³ , R ⁴ and X have the meanings given in Claim 1,
  and to the aldehyde of formula (VII) obtained in the sense of an aldol reaction, enzyme-catalyzed by means of a specific aldolase, dihydroxyacetone phosphate (DHAP) with CC linkage, with a typical sugar ring ring being cyclized to the ketone group of the DHAP unit via the previously generated nucelophilic Structure of formula (I) arises.