AU2008200567B2 - Synthetic Heparin Pentasaccharides - Google Patents

Description

00 TITLE 0 SSynthetic Heparin Pentasaccharides SField of the Invention 0 5 This invention is directed to intermediates, and processes for the chemical synthesis of AT-III binding heparin or heparinoid, pentasaccharides.

NO

Background Art 0o 0 10 Vascular thrombosis is a cardiovascular disease indicated by the partial or cN- total occlusion of a blood vessel by a clot containing blood cells and fibrin. In arteries, it results predominantly from platelet activation and leads to heart attack, angina or stroke, whereas venous thrombosis results in inflammation and pulmonary emboli. The coagulation of blood is the result of a cascade of events employing various enzymes collectively known as activated bloodcoagulation factors. Heparin, a powerful anticoagulant has been used since the late 1930's in the treatment of thrombosis. In its original implementation, tolerance problems were noted and so reduced dosage was suggested to reduce bleeding and improve efficacy. In the early 1970's, clinical trials did indeed indicate acceptable tolerance was obtainable whilst still preserving antithrombotic activity. Unfractioned heparin (UFH) is primarily used as an anticoagulant for both therapeutic and surgical indications, and is usually derived from either bovine lung or porcine mucosa. Amongst the modern uses of unfractioned heparin are the management of unstable angina, an adjunct to chemotherapy and anti-inflammatory treatment, and as a modulation agent for growth factors and treatment of haemodynamic disorders.

In the late 1980's, the development of low molecular weight heparins (LMWHs) led to improvements in antithrombotic therapy. LMWHs are derived from UFH by such processes as; chemical degradation, enzymatic depolymerisation and y-radiation cleavage. This class of heparins has recently been used for treatment of trauma related thrombosis. Of particular interest is the fact that their relative effects on platelets are minimal compared to heparin, providing an immediate advantage when treating platelet 00 compromised patients. The degree of depolymerisation of UFH can be

O

Scontrolled to obtain LMWH of different lengths. Dosage requirements for the Streatment of deep vein thrombosis (DVT) are significantly reduced when L employing LMWH as opposed to UFH, although in general the efficacy of both therapeutics seems to be comparable. In addition, LMWH can be effective as an alternative therapeutic for patients who have developed a sensitivity to UFH. Unfortunately, there has recently been a great deal of concern in the n use of LMWH due to the perceived potential for cross-species viral contamination as a result of the animal source of the parent UFH.

00 10 One way of avoiding the possibility of cross-species contamination, is Sto prepare heparins by chemical synthesis. This method would also provide the opportunity to develop second generation heparins or heparinoids, that can be tailored to target particular biological events in the blood coagulation cascade.

An investigation to determine the critical structural motif required for an important binding event in a coagulation cascade involving heparin, dates back to the 1970's. Some structural features of heparin were defined, but the binding domains of interest remained essentially undefined. Research conducted by Lindahl and co-workers [Lindahl, et al., Proc. Natl. Acad. Sci.

USA, 1980, Vol. 77, No. 11, 6551-6555; Reisenfeld, et al., J. Biol. Chem., 1981, Vol. 256, No. 5, 2389-2394] and separately by Choay and co-workers [Choay, et al., Annals New York Academy of Sciences, 1981, 370, 644-649] eventually led to the determination that a pentasaccharide sequence constituted the critical binding domain for the pro-anticoagulant cofactor, antithrombin III (AT-III). After determination of the critical heparin sugar sequence, complete chemical syntheses were embarked upon to further prove the theories. Complete syntheses of the pentasaccharide binding domain were completed at similar times by Sinay and co-workers and by Van Boeckel and co-workers [Sinay, et al., Carbohydrate Research, 132, (1984), C5-C9].

Significant difficulties were encountered during both these reported syntheses. The synthesis by Van Boeckel and co-workers provided a method on reasonable scale (156mg's of final product) and with improved yields compared to the Sinay synthesis, but still only provided an overall yield of 00

O

O

0

O

0

D

qj fc

(N

0.22%, (compared with 0.053% for the Sinay synthesis). One particular problem encountered during the final deprotection, was the intermolecular reaction of the hemiacetal (the reducing end functionality of the sugar), which led to the formation dimers and trimers. To reduce the likelihood of this 5 occuring, an a-methyl glycoside of the pentasaccharide was synthesised. The structures of interest are represented in Figure 1, wherein I represents the hemiacetal form, and II represents the a-methylglycoside form.

.OSO;

coo Oso 3 OSO3 HO H R=S03

HO

OCH

3 D E F G H

II

Figure I As mentioned, studies have determined that the significant biological event in preventing thrombosis is the binding of a pentasaccharide sequence [Choay et al., Ann. NY Acad. Sci., 1981, 370, 644-649] of heparin, to heparin cofactor antithrombin III (AT-III). As well as pentasaccharide I, the important derivative II has also been prepared by total synthesis [Choay et. al., Biochem. Biophys.

Res. Commun., 1983, 116, 492-499]. Compound II has recently completed phase III clinical trials for the treatment of deep-vein thrombosis. The following 00 patents display some relevance to the present invention. Patent US 4,401,662 O claims composition of matter on the pentasaccharide AT-Ill binding sequence ,c of heparin as does US 4,496,550. Patents EP 0,084,999 and US 4,818,816 detail synthetic methodologies towards pentasaccharide I, and derivative II.

Object of the Invention It is an object of the invention to provide a synthetic preparation for n heparin pentasaccharides, and intermediates thereof, and to novel Sintermediates for heparin pentasaccharides, and to novel heparin oo 10 pentasaccharides.

C The present invention provides composition of matter of intermediates, and a process for the synthesis, of AT-III binding heparins and heparinoids.

What this entails is a stepwise synthetic process employing monosaccharide building blocks.

The nature of the AT-III binding pentasaccharide is such, that under cursory analysis of the individual monomeric units constituting the pentasaccharide, we note that each is distinct from the others. Secondly, we can see that there is an alternating stereospecificity in regard to the glycosidic linkages (Fig. 2).

a-linked p-linked a-linked P-linked oso;3 COO OSO I OS0 3 o 0 0 0 0 OH OH oL O S O3 O OHo

HO

E O C BA SHNSO3 OH HNSO3 OS3 NHSO3 Glucosamine Glucuronic acid Glucosamine Iduronic acid Glycosides and hemi- 2-N-sulphated and 2-N-sulphated, 2-O-sulphated acetal of Glucosamine, 6-O-sulphated 3-O-sulphated and both 2-N-sulphated and 6-O-sulphated 6-O-sulphated Fig. 2 In a synthesis, the difference evident in each block requires that each individual monomer used in the synthesis will need a different protecting 00 group pattern. In light of this, it is essential in the synthesis of the above 0 pentasaccharide that a protecting group strategy is carefully conceived. As 4, can be seen, the pentasaccharide displays O-sulphation, N-sulphation, there C are free hydroxyl groups, and there are stereospecific glycosidic linkages.

Therefore, a protection strategy is required such that sulphation can be effected at the required sites, whilst leaving some hydroxyl groups unsulphated (note that due to the chemical lability of N- and O-sulphates, n sulphation needs to be effected late in the synthesis), a protection strategy is required that assists in effecting the appropriate glycosidic linkage and a oo 10 protection strategy is required that enables the correct (in terms of regio- and stereoisomerism) glycosidic linkages to be formed. a-Glycosidic linkages are typically generated by the use of what are known as non-participating protecting groups, whilst p-linkages are effected by participating protecting groups. Some N- and O-participating and non-participating protecting groups are known to the art (the art being considered carbohydrate chemistry). It is also well known to the art that the kind of protecting groups employed can effect the reactivity of the building block. The culmination of these requirements are demonstrated in the exemplary building block in Fig. 3 below, which displays the kind of characteristics required to effect the synthesis of heparin oligosaccharides.

OR

4 J-0

OR

2

X

R

3 0

NR

1 Fig. 3, Exemplary Building Block C In exemplary building block C, X is a leaving group suitable of reacting with another monomer or acceptor, to form an interglycosidic linkage; R 1 is a non-participating amino protecting group so as to effect an a-linkage upon activation of X followed by coupling to an appropriate acceptor; R 2 and R 4 can be similarly protected to allow for eventual O-sulphation, whilst R 3 is required to be differentially protected so as to allow the formation of an acceptor 00 hydroxyl group to couple this block to the next in the chain. The building 0 blocks in Fig. 4 exemplify the kind of derivatised monosaccharides required to effect the synthesis of heparin AT-III binding pentasaccharides.

ORs R ORs ORs 0 0 o 0 0 0 eventually require O-sulphation, the protecting groups represented by 'R' IN RHO RLO RO R0 R R E D C B OR ARA 00 Fig. 4 The protecting groups represented by 'Rs' in Fig. 4 are sites that will eventually require 0-sulphation, the protecting groups represented by 'RH' need to be orthogonal to 'Rs' and represents sites that will eventually become hydroxyl groups. The substituents 'Xi' and 'X 2 represent leaving groups that are activated to react with another suitable protected building block to form a glycosidic linkage, and,in the case of X 1 may also be derivatised as alkyl glycosides or substituted with a group suitable to allow conjugation to a support for drug delivery. The 'RL' groups are protecting groups orthogonal to both 'Rs' and and represent sites through which chain elongation via glycosylation occurs. is representative of either a protected or latent carboxylate function. The 'RA' groups are non-participating amino protecting groups that enable a-linkages to be formed while the 'RB' groups may be either a participating or non-participating amino protecting group. There is another level of complexity to be added to the synthesis in as much as the protecting groups in blocks D and B that are indicated by the boxes, need to be such that they allow for the formation of a p-glycosidic linkage. This may require a two stage protection at the indicated sites, ie. a protection followed by deprotection and subsequent reprotection with a different protecting group.

The initial protection is required to effect the correct stereochemistry in a glycosylation, and second stage protection to allow for the correct sulphation pattern.

00 As is evident, the pentasaccharide can be constructed in a variety of 0 different ways; blocks B and A can be coupled, blocks E and D can be Scoupled, block C can be coupled to either, and the resulting dimer and trimer can finally be coupled to form the pentasaccharide. Alternatively, each block S 5 can be added sequentially and so on. There are a number of alternative coupling sequences that can be easily conceived and the choice made in regard to this, in itself, has a marked effect on the synthetic methodologies that will finally be employed, and therefore impacts on the overall success of 0 the synthesis.

00 10 In one aspect the invention provides for a monosaccharide building block in C the D-glucopyrano configuration, for the preparation of synthetic heparinoids, said building block of General Formula I, ORs ,o 0 F-S RA X1 General Formula I (Block A) Wherein X 1 includes but is not limited to: hydroxy, alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, or other suitable leaving group; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; other suitable groups will be known to those skilled in the art, RA includes but is not limited to: an azido function, an amine; an NH-Dde, NH- DTPM, NH-Fmoc, NH-Boc, NH-Cbz, NH-Troc, N-phthalimido; or, other such suitable protected amino functions known to those skilled in the art, RH is a benzyl or, substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH and RA can combine together to form a cyclic carbamate; RL includes but is not limited to: a H atom; a levulinoyl, chloroacetyl, 4acetoxybenzoyl, 4-acetamidobenzoyl, 4-azidobenzoyl, or other substituted 00 benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-

O

Sacetamidobenzyl or other such suitable substituted benzyl type protecting n group; y-aminobutyryl, 4-N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1- C- ylidene)ethylamino]-butyryl, 4-N-[1-(1,3-dimethyl-2,4,6(1 0 5 trioxopyrimidin-5-ylidene)methylamino]-butyryl, 4-N-Alloc-butyryl, 4-N-Fmocbutyryl, 4-N-Boc-butyryl type protecting groups allyloxycarbonyl, allyl ether, carbonate type protecting groups; or RL and Rsi can combine to form a benzylidene or substituted benzylidene ring.; or, other such suitable protecting Sgroups as known to those skilled in the art, and 00 Rs includes but is not limited to: 4-methoxyphenyl; substituted benzyl groups; C- alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl;; or, other suitable protecting groups as known to those skilled in the art.

Alternatively RL and Rs can combine to form a benzylidene or substituted benzylidene ring.

In a second aspect the invention provides for a monosaccharide building block in the L-idopyrano conformation, for the preparation of synthetic heparinoids, said building block of General Formula II,

REO

O OR RLO ORs General Formula II (Block B) Wherein X 2 includes but is not limited to: a hydroxyl group; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, or other suitable leaving group; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may 00 be alpha or beta; other suitable groups will be known to those skilled in the Sart, SRs is defined as in General Formula I, CRH is defined as in General Formula 1, RL is defined as in General Formula I, and RE includes but is not limited to: methyl, C2-05 alkyl; substituted alkyl; or, benzyl and substituted benzyl groups; other suitable groups will be known to 0 those skilled in the art. Or RH is selected from the group consisting of benzyl or substituted 00 10 benzyl protecting group, allyl, allyloxycarbonyl; Rs is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; RE is selected from the group consisting of methyl, C2-C5 alkyl; substituted alkyl,C 3

-C

5 alkenyl; or, benzyl and substituted benzyl groups; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or other such suitable substituted benzyl type protecting group; y-aminobutyryl, 4-N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamino]-butyryl, 4-N-[1- (1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, 4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; allyloxycarbonyl, allyl ether, carbonate type protecting groups; X2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, imidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta.

00 In a third aspect the invention provides for a monosaccharide building block in

O

Sthe L-idopyrano configuration, for the preparation of synthetic heparinoids, 0 said building block of General Formula III, RI-- ORH P RMO X 2 -0 RLO ORs O SGeneral Formula III (Alternate Block B) CO 00 Wherein X 2 is defined as in General Formula II, Rs is defined as in General Formula II, RH is defined as in General Formula I, RL is defined as in General Formula I, and RM includes but is not limited to a p-methoxyphenyl protecting group or other suitable oxidatively labile protecting group; a trityl group; or, other such suitable protecting groups as known to those skilled in the art.

In a fourth aspect the invention provides for a monosaccharide building block in the D-glucopyrano configuration for the preparation of synthetic heparinoids, said building block of General Formula IV, ORs General Formula IV (Block C) Wherein X 2 is defined as in General Formula II, RB includes but is not limited to: an azido function, an amine; an NH-Dde or NH-DTPM group; or other suitably protected amino functions as known to those skilled in the art, or Rs (adjacent Re )and RB can combine together to form a cyclic carbamate; 00

O

O RL is defined as in General Formula I, and )Rs (adjacent RB is is selected from the group consisting of 4-methoxyphenyl; L substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, or RS 4 and RB may be combined to form a cyclic carbamate; SRs (adjacent the oxygen) is selected from the group consisting of 4- 00 10 methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, Sand substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl; In a fifth aspect the invention provides for a monosaccharide building block in the D-glucuronate configuration for the preparation of synthetic heparinoids, said building block of General Formula V, O ORE ORp X General Formula V (Block D) Wherein X 2 is as defined in General Formula II, Rp includes but is not limited to: 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups; carbonate protecting groups; or, other suitable protecting groups as known to those skilled in the art.

RL is defined as in General Formula I, and RE is defined as in General Formula II.

12 00

O

0 In a sixth aspect the invention provides for a monosaccharide building block in 3 the D-glucopyrano configuration for the preparation of synthetic heparinoids, said building block of General Formula VI, ORs t- O RHO- 0 tfl Rei Q 0 0 0 General Formula VI (Block E) Wherein X 2 is as defined as in General Formula II, RB is defined as in General Formula IV, RH may be selected independently and are defined as in General Formula I, and Rs is defined as in General Formula I or, wherein RH (adjacent the ORs moiety) is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RH (adjacent the Rb moiety) is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or this RH and RB independently can combine together to form a cyclic carbamate; Rs is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl; and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl,or Rs5 and RH can be combined to form a cyclic acetal or ketal moiety; RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RH adjacent the RB) and RB can combine together to form a cyclic carbamate; 0 0 X2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, Strichloroacetimidoyl, phosphate and related phosphate ester type leaving Sgroups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta.

In a seventh aspect the invention provides for a monosaccharide building n block in the D-glucopyrano configuration for the preparation of synthetic Sheparinoids, said building block of General Formula VII, 00 O ORs RLO 0^- C~o 0 NH X 1 General Formula VII (Common Intermediate for Blocks A, C and E) Wherein X 1 is defined as in General Formula I, RL is defined as in General Formula I, and Rs is defined as in General Formula I.

RL and Rs may also together combine to form a benzylidene or substituted benzylidene ring or

X

1 is selected from the group consisting of hydroxy, alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, atbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or other such suitable substituted benzyl type protecting group; y-aminobutyryl, 00 4-N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamino]-butyryl, 4-N-[1-

O

S(1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, 1C 4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; Sallyloxycarbonyl, allyl ether, carbonate type protecting groups.

Rs is selected from the group consisting of 4-methoxyphenyl, 4n methoxybenzyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, Sand substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate 0 10 protecting groups; tert-Butyldiphenylsilyl; RL and Rs may also together combine to form an alkylidene, isopropylidene, benzylidene or substituted benzylidene ring.

In an eighth aspect the invention provides for a disaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula VIII, ORs REO ORs° O RA X1 RLO ORs General Formula VIII (Block B-A) Wherein X, is defined as in General Formula I, RH1 is defined as being selected from RH of General Formula I, with the addition that RH1 and RA can combine together to form a cyclic carbamate, RA is defined as in General Formula I, with the addition that RH1 and RA can combine together to form a cyclic carbamate Rs is defined as in General Formula I, RH is defined as in General Formula I, 00 RL is defined as in General Formula I, and

O

0 RE is defined as in General Formula II or Xi is selected from the group consisting of hydroxy, alkenyloxy, C alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, imidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems n or solid supports; and the stereochemistry may be alpha or beta; RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RHI is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH1 and RA can combine together to form a cyclic carbamate; RA is selected from the group consisting of an azido function, an amine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Troc, N-phthalimido, NH-Ac, NH-Allyloxycarbonyl; or RH1 and RA can combine together to form a cyclic carbamate; Rs (on block A) is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl or benzoyl, Rs (on block B) is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; 00 0 RE is selected from the group consisting of methyl, C2-C5 alkyl; 3 substituted alkyl,C 3 -C5 alkenyl; or, benzyl and substituted benzyl groups; S 5 RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or n other such suitable substituted benzyl type protecting group; y-aminobutyryl, 0 4-N-1 -(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamino]-butyryl, 4-N-[1 00 10 (1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, S4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; allyloxycarbonyl, allyl ether, carbonate type protecting groups.

In a ninth aspect the invention provides for a disaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula IX, ORs R&P OR,

RM

RA X 1 RO ORs General Formula IX (Alternate Block B-A) Wherein X, is as defined as in General Formula I, RA is defined as in General Formula XIII, RH1 is defined as in General Formula XIII, Rs is defined as in General Formula I, RL is defined as in General Formula I, and RM is defined as in General Formula III Or alternatively, 00 X, is selected from the group consisting of hydroxy, alkenyloxy,

O

0 alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, Simidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; r- SRA is selected from the group consisting of an azido function, an Samine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Troc, N-phthalimido, 00 10 NH-Ac, NH-Allyloxycarbonyl; or RH1 and RA can combine together to form a Scyclic carbamate; Rs (on block A) is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; Rs (on block B) is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RH1 is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RHI and RA can combine together to form a cyclic carbamate; RM is selected from a p-methoxyphenyl or p-methoxybenzyl protecting group or other suitable oxidatively labile protecting group; a trityl group; 00

O

0 or RM and RL are combined together to form an isopropylidene, benzylidene, Ssubstituted benzylidene, cyclohexylidene or other acetal or ketal protecting group; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type IV protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or o other such suitable substituted benzyl type protecting group; y-aminobutyryl, 00 10 4-N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamino]-butyryl, 4-N-[1- S(1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, 4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; allyloxycarbonyl, allyl ether, carbonate type protecting groups.

In a tenth aspect the invention provides for a disaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula X, O ORE RLO- 0 O RS10 O R/X op ORs General Formula X (Block D-C) Wherein X 2 is as defined in General Formula II, Rsi is defined as being selected from Rs of General Formula I, with the addition that Rsi and RB can combine together to form a cyclic carbamate.

RB is defined as in General Formula IV, with the addition that Rsi and RB can combine together to form a cyclic carbamate.

Rs is defined as in General Formula I, Rp are defined as in General Formula V, RL is defined as in General Formula I, and 00 RE is defined as in General Formula II or

O

SX2 is selected from the group consisting of hydroxy, alkenyloxy, :3 alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, L imidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; SRs is selected from the group consisting of 4-methoxyphenyl; 00 10 substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted Salkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl; Rsi is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, or Rsi and RB may be combined to form a cyclic carbamate; RE is selected from the group consisting of methyl, C2-05 alkyl; substituted alkyl,C 3 -C5 alkenyl; or, benzyl and substituted benzyl groups; RB is selected from the group consisting of an azido function, an NH-Dde or NH-DTPM group, or Rs 4 and Re can combine together to form a cyclic carbamate; Rp (adjacent O-RL) is selected from the group consisting of 4methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; 00 RP (adjacent the link to block C) is selected from the group consisting of hydroxy, 4-methoxyphenyl;benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl;

NO

RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type 00 10 protecting group; a 4-acetoxybenzyl, 4-acetamidobenzyl or other such suitable substituted benzyl type protecting group; y-aminobutyryl, 4-N-[1 -(4,4-dimethyl- 2,6-dioxocyclohex-1-ylidene)ethylamino]-butyryl, 4-N-[1-(1,3-dimethyl- 2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, 4-N-Allocbutyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; allyloxycarbonyl, allyl ether, carbonate type protecting groups.

In an eleventh aspect the invention provides for a disaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula XI, ORs 0 ORE ORp X2 General Formula XI (Block E-D) Wherein X 2 is as defined in General Formula II, Rp are defined as in General Formula V, RE is defined as in General Formula II, 00 RB is defined as in General Formula IV, with the addition that RB and RH2 can

O

Scombine to form a cyclic carbamate, RH2 is defined as being selected from RH of General Formula I, with the addition that RB and RH2 can combine to form a cyclic carbamate, RH is defined as in General Formula I, and Rs is defined as in General Formula I or X2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, n trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting 0 10oo group; and the stereochemistry may be alpha or beta; Rp adjacent the O link) is selected from the group consisting of 4methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; Rp (adjacent X2) is selected from the group consisting of 4methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl; RE is selected from the group consisting of methyl, C2-05 alkyl; substituted alkyl, C3-C5 alkenyl; or, benzyl and substituted benzyl groups; RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RH2 and RB1 can combine together to form a cyclic carbamate; RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RH2 is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH2 and RBI independently can combine together to form a cyclic carbamate; 00

O

O

Io Rs is selected from the group consisting of 4-methoxyphenyl; 4- C methoxybenzyl, substituted benzyl groups; alkylacyl, benzoyl arylacyl or alkylarylacyl, and substituted alkylacyl, 4-chlorobenzoyl ,arylacyl or alkylarylacy protecting groups; allyloxycarbonyl, ethoxycarbonyl, tertbutoxycarbonyl,carbonate protecting groups; a tbutyldiphenylsilyloxy or n other such substituted silyloxy protecting group allyl, methoxymethyl, Smethoxyethyl, benzyloxymethyl; 00 Sor Rs and RH can be combined to form a cyclic acetal or ketal moiety; In a twelfth aspect the invention provides for a disaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula XII, ORs RHO 0 0 ORp General Formula XII (Alternate Block E-D) Wherein X 2 is as defined in General Formula II, Rp are defined as in General Formula V, RM is defined as in General Formula III, RB and RH2 are as defined in General Formula XI, RH isdefined as in General Formula I, and Rs is defined as in General Formula I or X2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving 00 groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting

O

0 group; and the stereochemistry may be alpha or beta; Rp (adjacent the O linking group) is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups; carbonate protecting groups; Rp (adjacent X) is selected from the group consisting of 4- (-i 00 10 methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and Salkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting (-i groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl; RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RH2 and RBI can combine together to form a cyclic carbamate; RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RH2 is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH2 and RBI independently can combine together to form a cyclic carbamate; Rs is selected from the group consisting of 4-methoxyphenyl; 4methoxybenzyl, substituted benzyl groups; alkylacyl, benzoyl, arylacyl or alkylarylacyl, and substituted alkylacyl, 4-chlorobenzoyl, arylacyl or alkylarylacy protecting groups; allyloxycarbonyl, ethoxycarbonyl, tertbutoxycarbonyl, carbonate protecting groups; is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such 00 substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, Sbenzyloxymethyl; SOr Rs and RH can be combined to form a cyclic acetal or ketal moiety; RM is selected from the group consisting of a p-methoxyphenyl protecting group or other suitable oxidatively labile protecting group; a trityl group.

00 SIn a thirteenth aspect the invention provides for a disaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula XIII,

A

RpO o 0 Oo 0 Rs1/ X2 ORs General Formula XIII (Alternate Block D-C) Wherein X 2 is defined as in General Formula II, RE and Rsi are defined as in General Formula X, Rs is defined as in General Formula I, Rp is defined as in General Formula V, and A includes but is not limited to; H, Methoxy, Methyl; other suitable substituents will be known to those in the art,or

X

2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, imidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta; 00 RB is selected from the group consisting of an azido function, an 0 amine; an NH-Dde or NH-DTPM group, or Rs 4 and RB can combine together Sto form a cyclic carbamate; Rs is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting I groups; a t butyldiphenylsilyloxy or other such substituted silyloxy protecting 0 group allyl, methoxymethyl, methoxyethyl benzyloxymethyl; 00 Rsi is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; or RS 4 and RB may be combined to form a cyclic carbamate; Rp (adjacent the O linking atom to the benzyl) is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; Rp (adjacent the O linking atom to C) is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl; A includes but is not limited to; H, Methoxy, Methyl; other suitable substituents will be known to those in the art.

00 In a fourteenth aspect the invention provides for a trisaccharide building block O for the preparation of synthetic heparinoids, said building block of General SFormula XIV, ORs

RHRO

A^ 0 ORE R -f RR OORs S5

OR

00 SGeneral Formula XIV (Block E-D-C) Wherein X 2 is defined as in General Formula II, RB and Rsi are defined as in General Formula X, Rs is defined as in General Formula I, Rp is defined as in General Formula V, RE is defined as in General Formula II, RB1 is defined as being selected from RB of General Formula IV, with the addition that RB1 can combine together with RH2 to form a cyclic carbamate, RH2 is defined as being selected from RH of General Formula I, with the addition that RH2 can combine together with RBlto form a cyclic carbamate, and RH is defined as in General Formula I or

X

2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta; Rp (adjacent block E) is selected from the group consisting of 4methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; 00 Rp (adjacent Block C) is selected from the group consisting of 4-

O

Omethoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and n alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting L. groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl; RE is selected from the group consisting of methyl, C 2

-C

5 alkyl; I substituted alkyl, C3-05 alkenyl; or, benzyl and substituted benzyl groups; 00 10 RBI is selected from the group consisting of an azido function, an 0 amine; an NH-Dde or NH-DTPM group, or RH2 and RBI can combine together to form a cyclic carbamate; RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RH2 is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH2 and RB1 independently can combine together to form a cyclic carbamate; Rs (on block E) is selected from the group consisting of 4methoxyphenyl; 4-methoxybenzyl, substituted benzyl groups; alkylacyl, ben:oyl, arylacyl or alkylarylacyl, and substituted alkylacyl, 4-chlorobenzoyl, arylacyl or alkylarylacy protecting groups; allyloxycarbonyl, ethoxycarbonyl, tertbutoxycarbonyl, carbonate protecting groups; is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; or Rs and RH can be combined to form a cyclic acetal or ketal moiety; 00 Rs (on block C and adjacent the ring O is selected from the group

O

Sconsisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting Sgroups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl;

NO

In Rs (on block C and adjacent the O linking atom) is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, oo 10 arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy Sprotecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; or Rs and RB may be combined to form a cyclic carbamate; RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or Rs 4 and RB can combine together to form a cyclic carbamate; In a fifteenth aspect the invention provides for a trisaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula XV, ORs ORs REO OR R0 O OR RLO- 0' f-RA

X

Rs0\l Re O ORs General Formula XV (Block C-B-A) 00 Wherein X 1 is defined as in General Formula I SRA and RH1 are defined as in General Formula VIII, SRs is defined as in General Formula I, jRH is defined as in General Formula I, RE is defined as in General Formula II, RB and Rsl are defined as in General Formula X, and RL is defined as in General Formula I or Xi is selected from the group consisting of hydroxy, alkenyloxy, 0 alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, 00 10 imidoyl, phosphate and related phosphate ester type leaving groups, a t 'butyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RHi is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RA is selected from the group consisting of an azido function, an amine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Cbz, NH-Troc, Nphthalimido, NH-Ac, NH-Allyloxycarbonyl; or RH and RA can combine together to form a cyclic carbamate; Rs on block A is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; Rs on block B is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate 00 protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy 0 0 protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl;

(N

Rs on block C is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl; 00 RS4 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, or RS 4 and RB may be combined to form a cyclic carbamate; RE is selected from the group consisting of methyl, C2-C5 alkyl; substituted alkyl,C3-C 5 alkenyl; or, benzyl and substituted benzyl groups; RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RS 4 and Re can combine together to form a cyclic carbamate; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or other such suitable substituted benzyl type protecting group; y-aminobutyryl, 4-N-[1 -(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamino]-butyryl, 4-N-[1- (1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, 4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; allyloxycarbonyl, allyl ether, carbonate type protecting groups; 31 00

O

(N

In a sixteenth aspect the invention provides for a tetrasaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula XVI, ORs n O REO O 0O RA and R RA X Rp is as defined in General Formula V, and 00 0 00

OORO

RO ORs

(N

General Formula XVI (Block D-C-B-A) Wherein X 1 is defined as in General Formula I RA and RHI are defined as in General Formula VIII, RL is defined as in General Formula I, RH is defined as in General Formula I, RE is defined as in General Formula II, RB and Rsl are defined as in General Formula X, Rp is as defined in General Formula V, and RL is as defined in General Formula I or Xi is selected from the group consisting of hydroxy, alkenyloxy, alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, imidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta, RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; 00 RH1 is selected from the group consisting of benzyl or substituted

O

Sbenzyl protecting group, allyl, allyloxycarbonyl, or RH1 and RA can combine Stogether to form a cyclic carbamate; 5 RA is selected from the group consisting of an azido function, an amine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Cbz, NH-Troc, Nphthalimido, NH-Ac, NH-Allyloxycarbonyl; or RHI and RA can combine Q together to form a cyclic carbamate, 00 10 Rs on block A) is selected from the group consisting of 4- Smethoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, Rs on block B) is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, Rs on block C and adjacent the ring oxygen) is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, Rs on block C and adjacent the linking O is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or 00 other such substituted silyloxy protecting group allyl, methoxymethyl,

O

0 methoxyethyl, benzyloxymethyl, :3 or RS 4 and RB may be combined to form a cyclic carbamate, S 5 RE on block D) is selected from the group consisting of methyl, C2alkyl; substituted alkyl, C3-C5 alkenyl; or, benzyl and substituted benzyl groups; SRE on block B is selected from the group consisting of methyl, 00 10 C2-C5 alkyl; substituted alkyl,C 3 -C5 alkenyl; or, benzyl and substituted benzyl Sgroups, RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or Rs4 and Re can combine together to form a cyclic carbamate, Rp adjacent RL) is selected from the group consisting of 4methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; Rp on group D and adjacent group C is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or other such suitable substituted benzyl type protecting group; y-aminobutyryl, 4-N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethylamino]-butyryl, 4-N-[1- 00 (1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, S4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; :3 allyloxycarbonyl, allyl ether, carbonate type protecting groups;

CD

In an seventeenth aspect the invention provides for a tetrasaccharide building block for the preparation of synthetic heparinoids, said building block of General Formula XVII, 00 ORs

SRHOO

RH

2 0 R ORE ORs REO R BR 0 0 ORp O R O Rs O0 ORs General Formula XVII (Block E-D-C-B) Wherein X 2 is defined as in General Formula IV, RH is defined as in General Formula I, RE is defined as in General Formula II, RB and Rsi are defined as in General Formula X, Rs is defined as in General Formula I, Rp is defined as in General Formula V, RL is defined as in General Formula I, and

RB

1 and RH2 are defined as in General Formula XIV, or RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RH2 is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH2 and RBI independently can combine together to form a cyclic carbamate; 00 Rs on block B is selected from the group consisting of 4-

O

O methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, n and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; Rs on block C and adjacent the ring O )is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl 0 10 or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting Sgroups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl; Rs on block C and adjacent the O linking atom) is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, or this Rs and RB may be combined to form a cyclic carbamate, Rs on block E) is selected from the group consisting of 4methoxyphenyl; 4-methoxybenzyl, substituted benzyl groups; alkylacyl, benzoyl, arylacyl or alkylarylacyl, and substituted alkylacyl, 4-chlorobenzoyl, arylacyl or alkylarylacy protecting groups; allyloxycarbonyl, ethoxycarbonyl, tertbutoxycarbonyl, carbonate protecting groups; is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl;or this Rs and RH can be combined to form a cyclic acetal or ketal moiety; 00 RE on block D is selected from the group consisting of methyl,

O

C

2

-C

5 alkyl; substituted alkyl, C3-C5 alkenyl; or, benzyl and substituted benzyl n groups; RE (on block B is selected from the group consisting of methyl, C 2 alkyl; substituted alkyl,C 3

-C

5 alkenyl; or, benzyl and substituted benzyl groups; RB is selected from the group consisting of an azido function, an 00 10 amine; an NH-Dde or NH-DTPM group, or RS 4 and RB can combine together 0 to form a cyclic carbamate; RBi is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RH2 and RB1 can combine together to form a cyclic carbamate; Rp on block D adjacent block E is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; Rp on block D adjacent block C is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl; X2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta 00 In a eighteenth aspect the invention provides for a pentasaccharide building

O

C block for the preparation of synthetic heparinoids, said building block of n General Formula XVIII,

OR

SOORRE

R EO O 0 5 0 ORs

OO

SE D C B A CN General Formula XVIII (Block E-D-C-B-A) Wherein X 1 is defined as in General Formula I RA and RH1 are defined as in General Formula VIII, and RH1 can also be allyl and alloxycarbonyl or RA and RH1 can combine together to form a cyclic carbamate.

Rs is defined as in General Formula I, RH is defined as in General Formula I,or RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, RE is defined as in General Formula II, RB and Rsi are defined as in General Formula X, Rp is defined as in General Formula V, and may be benzyl, Rp (adjacent the link from D to C) may also be silyl protecting groups, carbamate protecting groups, C3-C5 alkenyl, and RB and RH2 are defined as in General Formula XIV or RH2 is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH2 and RBe independently can combine together to form a cyclic carbamate, Rsi on block C (Rs 4 in the claims) is selected from the group consisting of 4methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate 00 protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy

O

O protecting group; allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, 03 or Rs 4 and Re may be combined to form a cyclic carbamate; Rs on block E C B A, (Rsi1 2 35 in the claims) can be a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, 00 In a nineteenth aspect, the invention provides a method for the preparation of compounds of the eighth aspect, involving the step of reacting a compound of the second or third aspect with a compound of the first or seventh aspect to form a new glycosidic bond.

In a twentieth aspect, the invention provides a method for the preparation of compounds of the eighth aspect, involving the step of selectively removing the protecting group RM from compounds of the ninth aspect and oxidizing the product of said deprotection.

In a twenty first aspect, the invention provides a method for the preparation of compounds of the tenth aspect, involving the step of reacting a compound of the fifth aspect with a compound of the fourth or seventh aspect to form a new glycosidic bond.

In a twenty second aspect, the invention provides a method for the preparation of compounds of the eleventh aspect, involving the step of reacting a compound of the fifth aspect with a compound of the sixth or seventh aspect to form a new glycosidic bond.

In a twenty third aspect, the invention provides a method for preparation of compounds of the thirteenth aspect involving the reaction of a compound of 00 the fourth or seventh aspect with a suitable donor molecule, to form a new 0 Sglycosidic bond.

In a twenty fourth aspect, the invention provides a method for the preparation of compounds of the fourteenth aspect involving the step of using any one or more of the compounds of the fourth, fifth, sixth, seventh, tenth, eleventh, twelfth or thirteenth aspect in a glycosidic bond forming reaction.

In a twenty fifth aspect, the invention provides a method for the preparation of oo 10 compounds of the fifteenth aspect involving the step of using any one or more Scompounds of the first, second, third, fourth, seventh, eighth and ninth aspects in a glycosidic bond forming reaction.

In a twenty sixth aspect, the invention provides a method for the preparation of compounds of the sixteenth aspect involving the step of using any one or more of the compounds of the first, second third, fourth, fifth, seventh, eighth, ninth, tenth, thirteenth or fifteenth aspect in a glycosidic bond forming reaction.

In a twenty seventh aspect, the invention provides a method for the preparation of compounds of the seventeenth aspect involving the step of using any one or more of the compounds of the second, third, fourth, fifth, seventh, tenth, eleventh, twelth, thirteenth or fourteenth aspect in a glycosidic bond forming reaction.

In a twenty eighth aspect, the invention provides a method for the preparation of compounds of the eighteenth aspect involving the step of using any one or more of the compounds of the 1,2,3,4,5,7, 8, 9, 10, 11, 12, 13, 14, or 15, 16 or 1 7 th aspect in a glycosidic bond forming reaction.

Best Mode Embodiments of the invention will be described with reference to the following examples: Standard operating protocols (SOP's) are provided for many of the examples.

List of Abbrevations: AcO: Acetyl, 00 All: Allyl, Alloc: Allyloxycarbonyl, Bn: Benzyl, Bz: Benzoyl, 5 CAN: (NH4) 2 Cev(N0 3 6 ceric ammonium (IV) nitrate, CIAc: Monochioroacetyl, Cres: p-TolyI, INO DCC: Dicyclohexylcarbodiimide, Dde: 1 -(4,4-dimethyl-2,6-dioxocyclohex-ylidene)ethyl, N 10 DEAD: Diethyl azodicarboxylate, 00 0 DIPEA: Diisopropylethylamine, 0 DMAP:4-NN-dimethylaminopyridine, DMF: NN-Dimethylformamide, DMTST: Dimethyl (methylthio)sulfoniumtetrafluoromethansulfonate, DTPMB: 2,6-d i-tert-butyl-4-m ethyl pyrid ine DTPM: (1 ,3-dimethyl-2,4,6 (1 H, 3H, 5H)-trioxopyrimidin-5-ylidene) methyl, Lev: 4-Oxopentanoyl, MCPBA: 3-chloroperbenzoic acid, Mes: Methanesulfonyl, Mp: 4-Methoxyphenyl, Mpm: 4-methoxybenzyl, NBS: N-Bromosuccinimide, NIS: N-Iodosuccinimide, NMP: N-Methylpyrollidone NPht: N-Phthaloyl PDC: Pyridiniumdichromate, Pent: n-Pentenyl, Ph 3 1P: Triphenyiphosphine, Piv: Pivaloyl, TBAF: Tetrabutylammoniumfluoride, TBD MS: tert-Butyld imethylsilyl, TBD1PS: tert-Butyldiphenylsilyl, TCA: Trichiloroacetim idyl, TEMPO: 2,2,6,6-Tetramrnethyl- 1-piperidinyloxyl, TFA: Trifluoroacetic acid, 00 TFAA: Trifluoroacetic acid anhydride,

O

0 Tf: Trifluoromethanesulfonyl, (-i STfN 3 Trifluoromethanesulfonyl azide, prepared from NaN 3 and Tf 2 0, T TfOH: Trifluoromethanesulfonic acid, THF: Terahydrofuran, TMS: Trimethylsilyl, Tos: p-Toluenesulfonyl, n p-TosOH: p-Toluenesulfonic acid, Trit: Triphenylmethyl.

00 SStandard Operating Procedures Standard Operating Procedure 1: Formation of Benzylidene acetals Standard Operating Procedure 2: Formation of p-Methoxybenzylidene acetals Standard Operating Procedure 3: Formation of isopropylidene acetals: Standard Operating Procedure 4: Dealkylidenation (Removal of isopropylidene, benzylidene and p-methoxybenzylidene) Standard Operating Procedure 5: Regioselective opening of the p-methoxybenzyliden acetal to a 6-O-pMethoxybenzyl ether Standard Operating Procedure 6: Regioselective opening of a benzylidene ring to a 4-O-benzyl ether Standard Operating Procedure 7: Introduction of a benzyl or p-methoxybenzyl ether Standard Operating Procedure 8:lntrodution of a tert-butyldiphenylsilyl ether Standard Operating Procedure 9: Cleavage of a tert-Butyl-diphenylsilyl ether Standard Operating Procedure 10:Introduction of a N-DTPM-group Standard Operating Procedure 11:Cleavage of a N-DTPM-group Standard Operating Procedure 12: Introduction of an azide group via diazo transfer reaction Standard Operating Procedure 13: Hydrolysis of thioglycosides (NBS) Standard Operating Procedure 14: Hydrolysis of thioglycosides (NIS) Standard Operating Procedure 15:Chemoselective Oxidation to Uronic acids Standard Operating Procedure 16: Methyl ester formation on the Uronic acids Standard Operating Procedure 17: Regioselective 6-O-Benzoylation Standard Operating Procedure 18: Common procedure for O-Benzoylation 0 0 Standard Operating Procedure 19:Common procedure for O-Acetylation

O

SStandard Operating Procedure 20: PDC-oxidation of alcohols to carboxylic n acids L. Standard Operating Procedure 21:Chemoselective 1-O-Benzoyl cleavage Standard Operating Procedure 22: Deacylation under Zemplen conditions Standard Operating Procedure 23: Introduction of the 4-Oxopentanoyl (=Levulinoyl) group q n Standard Operating Procedure 24:Cleavage of the 4-Oxopentanoyl Levulinoyl) group oo 10 Standard Operating Procedure 25: Formation of Trichloroacetimidates SStandard Operating Procedure 26: Regioselective introduction of a pMethoxyphenyl group under Mitsunobu conditions Standard Operating Procedure 27: Cleavage of the p-Methoxyphenyl ether Standard Operating Procedure 28: Cleavage of p-Methoxybenzyl ethers Standard Operating Procedure 29: Formation of a 2,3-cyclic carbamate Standard Operating Procedure 30: Cleavage of the N-phthaloyl group Standard Operating Procedure 31: Introduction of a thiocresyl ether at the reducing end Standard Operating Procedure 32: Glycosylation with thioglycosides a) NIS-promoted glycosylation b) DMTST promoted glycosylations: Standard Operating Procedure 33: Glycosylations with trichloroacetimidates Standard Operating Procedure 34: Glycosylations using 2,3-cyclocarbamoyl protected pThiocresyl glycosides as glycosyl donors Standard Operating Procedure 35: Introduction of an Alloc-group Standard Operating Procedure 36: Cleavage of an Alloc-group Standard Operating Procedure 37: Lewis acid mediated benzylation Standard Operating Procedure 38: benzylation under mild basic conditions Standard Operating Procedure 39: Ester cleavage under very mild conditions Standard Operating Procedure 1: Formation of Benzylidene acetals The starting material (47.5 mmol) was dissolved in acetonitrile (100 200 ml) and reacted with benzaldehyde dimethyl acetal (1.2 equiv.) and a catalytic amount of p-toluenesulphonic acid monohydrate (0.01-0.1 equiv). The 00 reaction was stirred at 500C under reduced pressure (350 mbar) until the TLC

O

shows completion. Subsequently, the mixture was neutralized with f triethylamine (pH 9) and concentrated in vacuo. The remaining residue was dissolved in an organic solvent dichloromethane or ethyl acetate) and extracted with H 2 0, saturated brine solution, dried over Na 2

SO

4 and concentrated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 70 and 95 0o S 10 Standard Operating Procedure 2: Formation of p-Methoxybenzylidene c- acetals The starting material (47.5 mmol) was dissolved in DMF/acetonitrile 100 200 ml) and reacted with p-methoxybenzaldehyde dimethyl acetal (1.2 equiv.) and a catalytic amount of p-toluenesulphonic acid monohydrate (0.01- 0.1 equiv). The reaction was stirred between 50 60 °C under reduced pressure (350 mbar) until the TLC shows completion. Subsequently, the mixture was neutralized with triethylamine (pH a 9) and concentrated in vacuo. The remaining residue was dissolved in an organic solvent (e.g.

dichloromethane or ethyl acetate) and extracted with H 2 0, saturated brine solution, dried over Na 2

SO

4 and concentrated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 70 and 85 Standard Operating Procedure 3: Formation of isopropylidene acetals: A solution of starting material (10 mmol) and catalytic amounts of camphorsulfonic acid (0.01-0.1 equiv) in 2,2-dimethoxypropane (50 ml) was stirred at 25 oC until completion, neutralized with triethylamine and concentrated. The remaining residue was dissolved in an organic solvent (e.g.

dichloromethane or ethyl acetate) and extracted with H 2 0 and saturated brine solution. The organic layer was dried over Na 2

SO

4 and concentrated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between and 93 00 0 Standard Operating Procedure 4: ,n Dealkylidenation (Removal of isopropylidene, benzylidene and p- A methoxybenzylidene) A solution of the acetal (31 mmol) in 150 ml dichloromethane was cooled to O°C and reacted with 80 aqueous TFA (20.0 ml, cooled to After stirring at 0°C until completion, the reaction mixture was neutralized with 30 SNaOH solution and extracted with water and saturated brine solution. The organic layer was dried over Na 2

SO

4 and concentrated. Final purification was Sachieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 70 and 95 Modification using p-TosOHxOH 2 in MeOH/CH 3 CN for cleavage: The acetal (16.6 mmol) was dissolved in 100mL of dry acetonitrile and 25 mL MeOH and the solution was reacted with catalytic amounts of p-TosOHxOH 2 The reaction mixture was heated at elevated Temperature (between 40 and until completion and then neutralized with Et 3 N, concentrated in vacuo and purified either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 70 and 95 Standard Operating Procedure 5: Regioselective opening of the pmethoxybenzyliden acetal to a 6-O-pMethoxybenzyl ether A suspension of the starting sugar (10.2 mmol), molecular sieves 3A (6.5 g, freshly activated) and Na(CN)BH 3 (3.85 g, 58.2 mmol) in dry DMF (90 ml) was stirred for 1 hr at r.t. and cooled down to 0°C. Subsequently, a solution of TFA (11.2 mL, 143.9 mmol in 51 mL dry DMF) was added dropwise and stirring continued at 50 to 600C until completion of the reaction. The reaction mixture was cooled to 20 0 C, diluted with ethyl acetate and extracted with a saturated aqueous NaHCO 3 solution and filtered through a celite pad. The combined organic layers were washed with saturated brine solution, dried over MgSO 4 and concentrated. Final purification was achieved either by 00 crystallization or by silica gel chromatography. The typical yields for the O product formation varied between 70 and 90 Standard Operating Procedure 6: Regioselective opening of a benzylidene ring to a 4-O-benzyl ether A solution of the starting material (3.4 mmol) in 25 mL dichloromethane is cooled to 0°C and to it is added of a solution of BH 3 in THF (1 M, 34 ml) and a solution of Bu 2 BOTf in dichloromethane (1 M, 3.7 ml). The reaction is Sstirred at 0°C till completion and then quenched with 10 ml Et 3 N and 10 ml 00 10 MeOH, concentrated and coevaporated three times with toluene. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yield for the product formation varied between and 90 Standard Operating Procedure 7: Introduction of a benzyl or pmethoxybenzyl ether The starting material (40.2 mmol) was dissolved in dry N,N'dimethylformamide (100 mL) at 0°C and reacted with NaH (48.24 mmol, 1.2 eq per OH to be benzylated). Then benzyl bromide (1.1 eq per OH to be benzylated) was added dropwise and stirring continued at 0°C until completion. The same conditions were applied for the introduction of an allyl ether (Allylbromide served as allylating reagent).

The excess of NaH was neutralized by careful addition of acetic acid, followed by concentration of the reaction mixture in vacuo. The residue was dissolved in ethyl acetate and subsequently washed with water, 10 aqueous HCI solution, saturated aqueous NaHCO 3 solution, saturated brine solution, dried over Na 2

SO

4 and concentrated in vacuo. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yield for the product formation varied between 70 and 92 The same procedure was followed for the formation of the p-methoxybenzyl ether except that p-methoxybenzyl chloride was added to the reaction instead of benzyl bromide and the reaction was performed between 50 and 600C.

00 Standard Operating Procedure 8: 0 O Introdution of a tert-butyldiphenylsilyl ether n A mixture of the starting material (29.0 mmol) and imidazole (70.1 mmol) was L dissolved in 80 mL anhydrous DMF and heated to 55 OC. To the solution was added tert-butyldiphenylchlorosilane (8.30 mL, 31.9 mmol) and stirring continued at 55 °C until completion. The reaction mixture was then cooled to °C and quenched with aqueous NaHCO 3 solution. After concentration in n vacuo, the residue was taken up in ethyl acetate and the organic phase washed successively with water, 10% aqueous citric acid, water, saturated 00 10 brine solution, dried over Na 2

SO

4 and evaporated. Final purification was Sachieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 85 and 95 Standard Operating Procedure 9: Cleavage of a tert-Butyl-diphenylsilyl ether To a solution of the silyl ether (2.15 mmol) in 2.5 mL dry THF and acetic acid (3.44 mmol) was added 1M TBAF solution in THF (3.22 mL) and stirring continued till completion of the reaction. Subsequently, the reaction mixture was concentrated in vacuo. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 85 and 97 Standard Operating Procedure 10:Introduction of a N-DTPM-group To a solution of the starting amine (24.5 mmol) in methanol (60 ml) is added a solution of the DTPM reagent (5.43 g, 25.7 mmol) in methanol (60 ml) at OC. After completion of the reaction, the reaction mixture was concentrated in vacuo, taken up in dichloromethane, extracted with water and saturated brine solution, dried over MgSO 4 and evaporated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 85 and 97 Standard Operating Procedure 11:Cleavage of a N-DTPM-group The starting material (40.94 mmol) was dissolved in dry DMF (50ml) and reacted with ethylene diamine (20 ml) at room temperature until completion.

00 The reaction mixture was concentrated in vacuo and coevaporated with

O

0 toluene. The residue was suspended in CHC1 3 and filtered through a Celite Spad. The filtrate was evaporated and final purification of the residue was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 85 and 92 00 00 Standard Operating Procedure 12: Introduction of an azide group via Sdiazo transfer reaction Sa) Preparation of a trifluoromethansulfonylazide solution: L A solution of sodium azide (492mmol) in water (80mL) was prepared under

N

2 -atmosphere. To this stirred solution was added dichloromethane (100mL) at 0°C, followed by the addition of triflic anhydride (16.5 ml) over 10 min. The mixture was further stirred for 2 hours at 0°C, the organic layer was separated and the aqueous layer was extracted with dichloromethane (2x40 mL). The Scombined organic layers were washed with saturated, aqueous NaHCO 3 0 10 solution (80 mL), water (80 mL) and dried over Na 2

SO

4 After filtration, this c solution was directly used for the diazotransfer reaction.

b) Diazotransfer reaction: To a solution of the starting material (26.0 mmol) and 4-N,N'- (dimethylamino)pyridine (14.5g) in acetonitrile (100mL) was added dropwise TfN 3 -solution (85ml) at room temperature within 10 min. The reaction was stirred till complete conversion of the starting material into the product. The reaction mixture was concentrated in vacuo to 30 ml and suspended in chloroform. After filtration through a Celite pad, the filtrate was concentrated and the residue was purified by filtration through a short silica gel pad. The typical yields for the product formation varied between 85 and 95 Standard Operating Procedure 13: Hydrolysis of thioglycosides (NBS) The starting thioglycoside (33.4 mmol) was suspended in 240 ml Acetone and 18 ml of distilled water and stirred for 45 min at -200C. After addition of NBS (155 mmol) stirring was continued at -200C. After completion, the reaction was stopped by addition of NaS 2 03/NaHCO 3 (20 aqueous solution 1/1) and the mixture diluted with ethyl acetate, subsequently washed with water and saturated brine solution. The organic layer was dried over Na 2

SO

4 and concentrated in vacuo. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 75 and 90 0 0 Standard Operating Procedure 14: Hydrolysis of thioglycosides (NIS) SThe starting thioglycoside (33.4 mmol) was suspended in 240 ml Acetone and S18 ml of distilled water and stirred for 45 min at -200C. After addition of NIS L (56.8 mmol) and TMSOTf (2.84 mmol) stirring was continued until completion.

The reaction was stopped by addition of NaS 2 03 /NaHCO 3 (20 aqueous solution diluted with ethyl acetate and washed with water and saturated brine solution. The organic layer was dried over Na 2

SO

4 and concentrated in n vacuo. Final purification was achieved either by crystallization petroleum spirit/ ethylacetate) or by silica gel chromatography. The typical yields for the O 10 product formation varied between 79 and 92 Standard Operating Procedure 15: Chemoselective Oxidation to Uronic acids A solution of the starting material (20.0 mmol) in dichloromethane (141 ml) was cooled to 0 0 C and subsequently mixed with TEMPO (0.205 mmol in 12.8 ml dichloromethane), Aliquat 336 (N-methyl-N,N-dioctyl-l-octanaminium chloride) (12.8 ml of a 0.08 M solution in dichloromethane) and KBr (2.08 mmol in 4.17 ml H 2 0) and stirring continued at 0°C. After 5 mins, a suspension of Ca(OCI) 2 (43.6 mmol) and NaHCO 3 (43.6 mmol) in 135 ml H 2 0 was added within 15 mins to the reaction mixture and stirring at 0°C was continued till completion. The reaction was concentrated in vacuo and freeze dried. The crude residue was used as such for the next reactions.

Standard Operating Procedure 16: Methyl ester formation on the Uronic acids The crude residue of the oxidation to the uronic acid was dissolved in 50 ml Toluene and 50 ml Methanol and titurated with TMSCHN 2 -solution (2M in hexane) until completion. The reaction mixture was quenched with acetic acid to destroy excess of esterification reagent and evaporated in vacuo. Final purification was achieved by silica gel chromatography. The typical yields for the product formation varied between 65 and 80 over the steps oxidation and esterification.

Standard Operating Procedure 17: Regioselective 6-O-Benzoylation 00 The starting material (32.04 mmol) was dissolved in dry dichloromethane

O

mL) and dry pyridine (10 mL) and cooled down to 45 0 C. Benzoyl chloride S32.04 mmol) was added dropwise and stirring continued at 45 0 C till completion. The reaction was concentrated in vacuo and coevaporated with toluene three times. The remaining residue was dissolved in dichloromethane and washed with 10% aqueous citric acid solution, saturated aqueous NaHCO 3 solution and saturated brine solution, dried over Na 2

SO

4 and evaporated in vacuo. Final purification was achieved either by crystallization or by silica gel Schromatography. The typical yields for the product formation varied between 00 oO 75 and 94 Standard Operating Procedure 18: Common procedure for O- Benzoylation To a solution of the starting material (11.9mmol) and DMAP (13.6mmol) in 1,2-dichloroethane was added dropwise benzoylchloride (1.7g, 12.1mmol). at 0°C. The mixture was then left to stir until completion (dependent on the substrate between 20 to 550C). Subsequently, the reaction mixture was diluted with dichloromethane and washed with water, 5% NaHSO 4 solution, saturated aqueous NaHCO 3 solution and saturated brine solution. The organic layer was dried over MgSO 4 followed by removal of the solvent in vacuo to give a crude residue. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 80 and 96 Standard Operating Procedure 19:Common procedure for O-Acetylation To a suspension of the starting material (235 mmol, 3 acetylation sites) in pyridine (350 ml) at O°C was added dropwise acetic anhydride (175 ml). After completion of the addition, the reaction was allowed to return to room temperature and stirred until completion. The reaction mixture was evaporated to dryness and 3x coevaporated with toluene. The residue was taken up in dichloromethane and washed with 5 aqueous NaHSO 4 -solution, saturated aqueous NaHCO 3 -solution, water and saturated brine solution. The organic layer was dried over MgSO 4 and evaporated. Final purification of the residue 00 was achieved either by crystallization or by silica gel chromatography. The

O

0 typical yields for the product formation varied between 88 and 98 C Standard Operating Procedure 20: PDC-oxidation of alcohols to S 5 carboxylic acids The starting material (1.15 mol) was dissolved in anhydrous DMF (7.0 ml) and reacted with PDC (11.5 mmol) under stirring at room temperature until Q complete conversion into the uronic acid. The reaction mixture was Ssubsequently poured into 50 ml water and the whole extracted with diethyl 00 10 ether. The combined ether layers were washed with 10 aqueous citric acid Ssolution, filtered through a short silica gel pad, dried over MgSO 4 evaporated and dried under high vacuum.

Standard Operating Procedure 21:Chemoselective 1-O-Benzoyl cleavage The starting material (36.8 mmol) was dissolved in dry DMF (80 ml) and cooled to 0°C. Subsequently, hydrazine acetate (44.06 mmol) was added and stirring continued until completion. After addition of acetone and acetic acid the reaction mixture was concentrated in vacuo. The residue was dissolved in dichloromethane and extracted with 10% aqueous citric acid solution, saturated NaHCO 3 solution, water and saturated brine solution, dried over MgSO 4 evaporated and dried under high vaccuum. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 72 and 88 Standard Operating Procedure 22: Deacylation under Zemplen conditions The starting material (23.7 mmol) was suspended in dry MeOH (70 ml) and stirred for 30 mins at 0 OC. Subsequently, NaOMe (0.1 equiv. O-Acyl group) was added (positive flush of N 2 and stirring was continued at 00 C until completion. Finally, the reaction was neutralized with 10 aqueous HCI and the solvent evaporated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yield for the product formation varied between 90 and 98 00

O

0 Standard Operating Procedure 23: Introduction of the 4-Oxopentanoyl S(=Levulinoyl) group a) Preparation of the Lev 2 0 solution: To a solution of DCC (31.2 mmol) in 100 mL dichloromethane was added levulinic acid (62.4 mmol) and DIPEA (62.42 mmol). The supernatant was used as such for the l levulination reaction.

SReaction 00 10 The above Lev 2 0 solution was added to a solution of the starting sugar (15.6 Smmol) dissolved in 25 mL of dry dichloromethane and stirring was continued until completion. Subsequently, the reaction mixture was filtered through a Celite pad and all combined organic layers were extracted with 10 aqueous citric acid solution, saturated aqueous brine solution, dried with Na 2

SO

4 and concentrated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 85 and 96 Standard Operating Procedure 24:Cleavage of the 4-Oxopentanoyl Levulinoyl) group A solution of the starting sugar (1.28 mmol) and acetic acid (1.35 mL) in pyridine (5.0 mL) was cooled to 0 °C followed by addition of hydrazine hydrate (200 pL). Stirring at 0 °C was continued until completion and the reaction mixture diluted with dichloromethane, subsequently extracted with aqueous citric acid, 10 aqueous NaHCO 3 solution, saturated brine solution, dried over Na 2

SO

4 filtered and concentrated. Final purification was achieved either by crystallization or silica gel chromatography. The typical yields for the product formation varied between 80 and 95 Standard Operating Procedure 25: Formation of Trichloroacetimidates a) with DBU: A solution of the starting sugar (1.99 mmol) and trichloroacetonitrile (601 [L, 5.87 mmol) in 5 mL dry dichloromethane was stirred at room temperature for 00 30 min. The reaction mixture was then cooled to 0°C and DBU (100 pmol)

O

Sadded. Stirring was continued until completion (dependent on the substrate, stirring was performed from 0°C to 200C). The reaction mixture was concentrated to one half of its volume and directly loaded on a short plug of silica gel and purified via silica gel chromatography. The typical yields for the product formation varied between 78 and 95 b) with K 2

CO

3 A solution of the starting sugar (1.99 mmol) and trichloroacetonitrile (601 PL, 00 S 10 5.87 mmol) in 5 mL dry dichloromethane is stirred at rt for 30 min. The c- reaction mixture was then cooled down to 0°C and anhydrous K2CO3 (19.9 mmol) added. The reaction was stirred at 0°C till completion and then filtered through a celite pad. The filtrate was dried over Na 2

SO

4 and evaporated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yield for the product formation varied between 78 and 95 Standard Operating Procedure 26: Regioselective introduction of a p-Methoxyphenyl group under Mitsunobu conditions A solution of the starting sugar (13.52 mmol), 4-methoxyphenol (20.3 mmol) and triphenylphosphine (20.3 mmol) in 85 ml dry dichloromethane was stirred at 0°C for 45 min. After addition of DEAD-reagent (22.9 mmol) at 0°C, the reaction mixture was further stirred at room temperature until completion, filtered through a celite pad, diluted with dichloromethane and extracted with 10 aqueous NaHCO3/NaOH solution 10 aqueous citric acid solution and aqueous saturated brine solution. The organic layer was dried over Na 2

SO

4 and concentrated. Final purification was achieved by silica gel chromatography. The typical yield for the product formation varied between and 89 00 Standard Operating Procedure 27: Cleavage of the p-Methoxyphenyl 0 ether The starting material (1.18 mmol) was dissolved in 30 ml acetonitrile and ml water and cooled to 0°C. Subsequently, CAN (3.83 mmol) was added and stirring continued at 0°C until completion. The reaction mixture was diluted with ethyl acetate and extracted with water. The aqueous layer was made Salkaline by addition of solid NaHCO 3 and back extracted with ethyl acetate.

SThe combined organic layers were extracted with saturated aqueous NaHCO 3 00 10 solution and saturated brine solution, dried over MgSO 4 and evaporated.

Final purification was achieved by silica gel chromatography. The typical yields for the product formation varied between 73 and 89 Standard Operating Procedure 28: Cleavage of p-Methoxybenzyl ethers The starting material (0.60 mmol) was dissolved in 27 ml acetonitrile and ml water and cooled to 0°C. Subsequently, CAN (4.5 equiv.) was added and stirring continued from 0°C to room temperature until completion. The reaction mixture was diluted with ethyl acetate and extracted with water. The aqueous layer was made alkaline by addition of solid NaHCO 3 and back extracted with ethyl acetate. The combined organic layers were extracted with saturated aqueous NaHCO 3 solution and saturated brine solution, dried over MgSO 4 and evaporated. Final purification was achieved by silica gel chromatography.

The typical yields for the product formation varied between 73 and 85 Standard Operating Procedure 29: Formation of a 2,3-cyclic carbamate To a stirred solution of the starting material (3.56 mmol) in dichloromethane (100 ml) and 10% aqueous solution of NaHCO 3 (75 ml) was added a solution of triphosgene (1.25 mmol) in 10 ml dry dichloromethane. The reaction was stirred at room temperature till completion. The organic phase was washed with water, dried over Na 2

SO

4 filtered and concentrated. Final purification was achieved either by crystallization or silica gel chromatography. The typical yield for the product formation varied between 75 and 95 00 Standard Operating Procedure 30: Cleavage of the N-phthaloyl group O The N-phthaloylated starting material (45.9 mmol) was dissolved in n-butanol S(200 ml) and treated with 1,2-diaminoethane (50 ml) at 1000C. After stirring at l 1000C until completion, the reaction mixture was concentrated in vacuo, coevaporated with toluene three times and dried under high vacuum. Final purification was achieved by silica gel chromatography. The typical yield for the product formation varied between 78 and 92

\O

SStandard Operating Procedure 31: Introduction of a thiocresyl ether at oO 10 the reducing end SA solution of the 1-O-glycosyl acetate (10.48 mmol) and p-thiocresol (12.58 mmol) in dry dichloromethane (30ml) was stirred at 0°C and subsequently activated by the addition of boron trifluoride diethylether complex (12.58mmol) over 5 min. Stirring was continued (0°C 20°C) until completion and the reaction stopped by the addition of triethyl amine (14.0 mmol). The reaction mixture was diluted with dichloromethane and extracted with saturated NaHCO 3 -solution, water and saturated brine solution, dried over MgSO 4 and evaporated in vacuo. Final purification was achieved by crystallization or silica gel chromatography. The typical yield for the product formation varied between 81 and 92 Standard Operating Procedure 32: Glycosylation with thioglycosides a) NIS-promoted glycosylation A mixture of glycosyl acceptor (1 mmol), thioglycoside (1 mmol) and 1.0 g of freshly activated molecular sieves in 20 ml of a dry solvent CH3CN,

CH

2

CI

2 Toluene, Ether) was stirred for 45 min at r.t and cooled down to the reaction temperature. Subsequently, N-lodosuccinimide (1.7 mmol) was added and stirring continued for 20 min at the reaction temperature. After the addition of a Lewis acid as promotor TfOH, 85-170 pmol), stirring was continued at the reaction temperature until completion. The reaction mixture was quenched with triethyl amine, filtered through a celite pad and extracted with a 10 aqueous KHCO 3 /Na 2

S

2 03 solution, water and saturated brine solution, dried over MgSO 4 and evaporated. Final purification was achieved 00 by silica gel column chromatography. The typical yields for the product 0 Sformation varied between 65 and 85 .C b) DMTST promoted glycosylations: L A mixture of glycosyl acceptor (1 mmol), thioglycoside (1 mmol) and 1.0 g of freshly activated molecular sieves in 20 ml of a dry solvent CH3CN,

CH

2

CI

2 Toluene, Ether) was stirred for 45 min at r.t and cooled down to the reaction temperature. Subsequently, DMTST (3-5 equiv.) was added and VB stirring continued at the reaction temperature until completion. The reaction 0 mixture was quenched with triethyl amine, filtered through a celite pad and 00 10 extracted with aqueous NaHCO 3 -solution, water and saturated brine solution, O dried over Na 2

SO

4 concentrated in vacuo and purified by silica gel column chromatography. The typical yields for the product formation varied between and 85 Standard Operating Procedure 33: Glycosylations with trichloroacetimidates A suspension of the trichloroacetimidate (1.54 mmol), glycosyl acceptor (1.13 mmol) and freshly activated molecular sieves (1.0 g) in an anhydrous solvent

CH

3 CN, CH 2 01 2 Toluene, Ether, 20 mL) was stirred at rt for 1 h and then cooled to reaction temperature. Subsequently, a catalytic amount of a promotor TMSOTf, 0.01-0.1 equiv.) was added and stirring continued at reaction temperature until completion. The reaction was quenched with triethylamine) and filtered through a Celite pad. The combined organic layers were washed with aqueous NaHCO 3 -solution and saturated brine solution, dried over Na 2

SO

4 concentrated in vacuo and purified by silica gel column chromatography. The typical yields for the product formation varied between and 85 00 Standard Operating Procedure 34:

O

0 Glycosylations using 2,3-cyclocarbamoyl protected pThiocresyl Sglycosides as glycosyl donors PhSCI (0.2 mmol, 2 equiv.) in dry dichloromethane (1 ml) was added dropwise to a mixture of AgOTf (0.2 mmol) in dry dichloromethane (2 ml) at -780C containing freshly activated molecular sieves 3 A. After stirring for 15 mins at n -780C, a solution of the thioglycoside (0.1 mmol, 1 equiv.) and DTBMP (0.2 Smmol, 2 equiv.) in dry dichloromethane (2 ml) was slowly added. After further 00 10 stirring for 15 mins at -78 0 C, the glycosyl acceptor (0.2 mmol, 2 equiv.) in dry Sdichloromethane (1 ml) was slowly added and stirring continued until completion. The reaction was quenched with saturated aqueous NaHCO 3 solution (1 ml), warmed to rt and diluted with dichloromethane. The organic layer was dried over MgSO 4 filtered and evaporated. Final purification was achieved by silica gel chromatography. The typical yields for the product formation varied between 60 and 90 Standard Operating Procedure 35: Introduction of an Alloc-group A solution of starting material (2 mmol), dry pyridine (5 mmol) and dry THF ml) was cooled to OoC. Subsequently, Allylchloroformate (2.2 mmol) were added dropwise and stirring was continued until completion. The reaction mixture was diluted with dichloromethane and subsequently washed with aqueous citric acid solution, saturated NaHCO 3 solution, water and saturated brine solution. The organic layer was dried over Na 2

SO

4 filtered and evaporated. Final purification was achieved either by crystallization or by silica gel chromatography. The typical yields for the product formation varied between 80 and 95 Standard Operating Procedure 36: Cleavage of an Alloc-group A mixture of the Allyloxycarbonate (1.17 mmol), dimedone (1.33 mmol) and Pd(Ph 3

P)

4 (0.30 mmol) was dissolved in dry THF (60 ml) and stirred under Ar atmosphere until completion of the reaction. The reaction mixture was 00 concentrated in vacuo and purified by silica gel chromatography. The typical Syields for the product formation varied between 78 and 97

N

Standard Operating Procedure 37: Lewis acid mediated benzylation To a stirred mixture of the starting material (1 mmol) and benzyl trichloroacetimidate in dry hexane/dichloromethane (10 ml, 2/1) was added Q Lewis acid (0.01-0.05 equiv., e.g. TMSOTf, TfOH) and stirring was continued Sat rt until completion. The reaction was quenched with triethyl amine and 00 10 concentrated. Final purification was achieved by silica gel chromatography.

SThe typical yields for the product formation varied between 50 and 92 00 Standard Operating Procedure 38: benzylation under mild basic

C

Sconditions T. The starting material (3.49 mmol) was dissolved in dry DMSO (20 ml) and cooled to 0°C. To the stirred solution were added successively benzyl bromide equiv./OH-group), barium oxide (1.5 equiv/OH-group), catalytic amounts of TBAI 0.05 eqiv./OH-group) and potassium hydroxide (3.5 equiv./ OH- Sgroup). Stirring was continued from 0°C to rt until completion. The reaction Swas quenched with methanol, and further stirred for 30 min. After dilution with 00 10 ether, the organic layer was washed with water and brine solution, dried over SMgSO 4 and concentrated in vacuo. Final purification was achieved by silica gel chromatography.

Standard Operating Procedure 39: Ester cleavage under aqueous conditions The starting material (0.3mmol ester groups) was dissolved in 11.8 ml of a mixture of water and THF cooled to 0°C and reacted with 1M aqueous NaOH-solution (5.0 ml). Stirring was continued until completion and the reaction mixture titurated with 10 aqueous HCI-solution to a pH of 9.5. After evaporation of the THF, the mixture was freeze dried and the remaining residue purified by silica gel chromatography to yield the product. The typical yields for the product formation varied between 85 and 95 Example 1: Synthesis of Building Blocks A-1 and A-2 from N-Acetyl Glucosamine HO H a10 HO 0bO HO HO 0 ANH H~ AcHA-I-i AcNH Oe HO~ HO 0 HO 0 A-I- NHHO DTPMHN OMe A-1-i NH2A-I-iii e 0 0 0 0 RJD""

HO\

mkl R H: A-1-v-a H2N OlMe DTPMHN I R R A--iv-a R MeO: A-1-iv-b R MeO: A-I-v-b

M

f

O

e R H: A-I-vi R =MeO: A- BzO\

N,

i-a OQMe I -vi-bh

HO

>HO 0 A-I -viiH N3 OMe O~le R H: A-I-vii-a R MeO: A-I-vii-b HO 0 BnO 'j OlMe A-2 Example 1: Synthesis of building blocks A-I and A-2 from N-Acetyl glucosamine, yields are reported for R=H, conditions; a) Amberlite IR 120 (ion exchange resin) MeOH, 60 (70 b) 1IM NaOH, 12000; c) 1.

SOP 10; 2. Ac 2 O, pyridine; 3. NaOMe, MeOH (70 4 steps); d) SOP 1 (91 for R=H) or SOP 2 for R OMe; e) SOP 11, f) SOP 12, (85 2 steps); g) SOP 7, (91 h) SOP 4, (91 i) SOP 17, (82 SOP Preparation of A-I-i: 0 0 N-Acetyl-2-deoxy-a/3-D-glucopyranoside (8.5 g, 38.4mmol) was suspended in 0 100 ml dry methanol. Subsequently, 12.0 g Amberlite IR 120 iron exchange Sresin (H'-form) was added and the reaction mixture refluxed for 70 hrs at C 650C. After cooling to 25 oC, the iron exchange resin was removed by filtration and several times extracted with methanol. The combined methanol layers were neutralized with triethyl amine and concentrated in vacuo. The crude residue was purified by crystallization to furnish the title compound in 70 yield (ao/ -mixture).

00 0 10 Preparation of A-1-iii: c Methyl glycoside A-1-i (20.6 mmol) was suspended in 100 ml aqueous NaOH solution (1 M) and stirred under reflux at 1200C until completion. After cooling and neutralization with 10 aqueous HCI, the mixture was concentrated in vacuo and crude A-1-ii suspended in 200 ml methanol and reacted with N- [(1,3-dimethyl-2,4,6 (1H,3H,5H)-trioxopyrimidin-5-ylidene) methyl]-N',N"-dimethylamine (23.6 mmol) at 500C (pH 9.0) until completion. After evaporation and drying, crude A-1-iii was reacted with 150 mL acetylation mixture (pyridine/Ac 2 0, 2/1, v/v) until completion, concentrated in vacuo, coevaporated with toluene and dried. The residue was suspended in ethyl acetate and extracted with water, 10 aqueous HCI, saturated, aqueous NaHCO 3 solution and H 2 0, dried over Na 2

SO

4 and concentrated. The crude residue was dissolved in dry methanol and reacted with a catalytic amount of NaOMe.

After completion, the reaction was neutralized with Amberlite IR 120 and filtered. The organic layer was evaporated and dried to furnish the title compound A-1-iii in 70 yield (over 4 steps).

Preparation of A-1-iv-a: Methyl-2-deoxy-2-N-[1 -(1,3-dimethyl-2,4,6(1 H, 3H, ylidene) methyl]-a-D-glucopyranoside A-1-iii (16.0 g, 44.5mmol) in acetonitrile (200ml) was reacted with benzaldehyde dimethyl acetal (14.0 ml, 92.3 mmol) and a catalytic amount of p-toluenesulphonic acid monohydrate. After 2 hours at 550 C, the mixture was neutralized and evaporated. The remaining residue 00 was extracted, washed and evaporated. Yield: 18.3 g (92 Rf 0.20 (1,2-

O

0 dichloroethane/ ethylacetate 7/3).

Preparation of A-1-v-a: Methyl-4,6-O-benzylidene-2-deoxy-2-N-[1-(1,3-dimethyl-2,4,6(1H, 3H, methyl]-a-D-glucopyranoside A-1-iv-a (18.30 g, 40.90 mmol) in DMF (50ml) was reacted with ethylenediamine (20ml) at room temperature. After stirring for 35 minutes, the mixture was concentrated. Yield: S10.90 g (94.7 Rf 0.18 (chloroform/methanol 9/1).

00 C- Preparation of A-1-vi-a: To a solution of methyl-2-amino-4,6-O-benzylidene-2-deoxy-c-D-glucopyranoside (7.5 g, 26.7 mmol) and 4-N,N'-(dimethylamino)pyridine (14.5g) in acetonitrile (100mL) was added TfN 3 -solution (85ml) at room temperature.

The reaction mixture was concentrated and the residue was purified by filtration through a short silica gel pad. Yield: 7.00 g (85.3 Rf 0.18 (chloroform/ methanol 9/1).

Preparation of A-1-vii-a: Methyl 2-azido-2-deoxy-4,6-benzylidene-a-D-glucopyranoside A-1-vi-a (10.87g, 35.40mmol) in N,N'-dimethylformamide (50mL) was reacted with NaH (95 0.92g, 36.4 mmol) and benzyl bromide (5.47 ml, 45.9 mmol). After completion, the excess of NaH was quenched, followed by concentration. The residue was extracted, washed and concentrated. Yield: 12.93 g (92.0 Rf 0.37 (petroleum spirit/ethyl acetate 3/1).

Preparation of A-1: Methyl-2-azido-3-O-benzyl-2-deoxy-a-D-glucopyranoside (9.87g, 31.9 mmol) in dichloromethane (50 mL) and pyridine (10 mL) was treated with benzoyl chloride (3.72mL, 32.04 mmol) at 45 0 C for 2 hours. The reaction was concentrated and the residue extracted, washed and evaporated. Yield: 10.78 g (81.7 Rf 0.31 (petroleum spirit/ ethylacetate 1/1).

00 Compound A-1: 'H-NMR (400 MHz, CDCI 3 6= 8.03 2 H, Aryl), 7.57 (in, 1H, Aryl), 7.45- 7.29 (in, 7H, Ajyl), 4.93 1 H, Jge m 10.8 Hz, OCH 2 4.82 1IH, Jgem 10.8 Hz, OCH 2 4.81 d, 1IH, J1, 2 =3.6 Hz, H-1lcx), 4.73 (dd, 1IH, J 5 6 a 4.4 Hz, Jge.m 12.0 Hz, H-6a), 4.47 (dd, 1 H, J 5 6 b 2.0 Hz, H-6b), 3.85 dd, 1 H, J34= 8.8 Hz, 3.57 ddd, 1IH, J 4 5 10.0 Hz, 3.45 3H, OMe), 3.37 dd, 1IH, J 2 3 10.0 Hz, 2.80 (bs, 1IH, 4-OH).

00 00 Example 2: Synthesis of Building Blocks A-3 and A-4 R HO R Bn DTPMHN DTPMHN R H: A-1-iv-a R H: A--ix-a OMe R MeO: A-1-iv-b R MeO: A-1-ix-b SHO d 0I HO O BnO MpmO HO 1 00 DTPMHN OMe Bn SA-1-x

DTPMHN

C-i OMe BzO~ A-4 BnO

DTPMHN

OMe A-3 Example 2: Synthesis of building block A-3 and A-4, conditions: a) SOP 7, (72 for R b) SOP 4, (82 c) SOP 17, (84 d) SOP Compound A-3: 'H-NMR (400 MHz, CDCI 3 6= 10.16 (dd, 1 H, JNH,2= 9.4 Hz, J NH, =C-H 14.0 Hz, NH), 8.11 1H, 7.68-7.22 (3m, 8H, Aryl), 4.84 d, 1H, J 1 2 Hz, H-la), 4.83 (dd, 1H, J 6 a, 6b 12.3 Hz, J 5 6 a 3.5 Hz, H-6a), 4.73 1H, Jgem 11.7 Hz, OCH 2 4.46 (dd, 1H, J5.6b= 2.1 Hz, H-6b), 3.91 1H, 3.72 (dd, 1H, J 3 4 J2.3 8.8 Hz, 3.57 (ddd, 1H, J 4 5 9.5 Hz, 3.48 3H, OMe), 3.38 (ddd, 1H, J 2 3 10.5 Hz, 3.32 3H, NMe), 3.31 (s, 3H, NMe), 3.05 (bs, 1H, 4-OH).

Examp~le 3: Synthesis of L-ido configured cilycosyl donor B-I

HO

HO' OBn .0 b 0 ON I 0n MesO- MesO- OBn 0 B-1Iv AcO OBn $0 SCres AcO QAc

C

f e B-I -vi HO OBn -O SCres B-I viii OH OH OBn 0 Sres 0 OH Me 2 C 0 OBn HO OBn MPOOBn 0 SCres 'F 0rese r 1 0Jz S~res L -0~S~e :0 B Ix 1xi 0 OBz OH OBz OH OBz Me 2

C

MPOOBn OLev OBz Example 3: Synthesis of Building Block B-1, conditions: a) SOP 7, b) aqueous Acetic acid, 6000 c) Methanesulfonyl chloride, Pyridine, 0 0 C-fRT (87 d) Cesium Acetate, Ac 2 O, 12000 e) SOP 22, f) 1. 90% TEA, 000; 2. Ac 2 O, Pyridine; 3. SOP 31, 3 steps); g) SOP 22, h) SOP 3, i) SOP 18, j) 80% acetic acid, 10000C k) SOP 26, 1) SOP 23, 00

O

9 Preparation of B-1-iii: B-1-ii (15.60 mmol) was dissolved in 60 aqueous acetic acid (50 ml) and S 5 stirred at 60 °C until completion. After neutralization with solid NaHCO 3 the mixture was evaporated and co evaporated with toluene. Crude B-1-iii was dissolved in CHCI 3 /H20, the organic layer separated, dried over Na 2

SO

4 and tI evaporated. The remaining residue was purified by a short silica gel chroma- Stography to yield B-1-iii in 90 (4.36 g).

00 SPreparation of B-1-iv: (N 17.72 mmol of B-1-iii was dissolved in 25 ml dry pyridine, to which mesyl chloride (methylsulfonyl chloride, 42.5 mmol) was added dropwise at 0°C. The mixture was stirred at 4°C until completion and was subsequently poured into warm water (50°C, 90 ml), cooled and the precipitate isolated by filtration.

B-1-iv was obtained after drying in 87 yield (7.19 g).

Preparation of B-1-v: B-1-iv (6.43 mmol) and cesium acetate (64.3 mmol) were suspended in 25 ml acetic anhydride and refluxed at 125 °C until completion. The reaction mixture was concentrated in vacuo, co evaporated with toluene and the residue extracted from ethyl acetate/H 2 0 The organic layer was collected and washed with saturated aqueous NaHCO 3 solution and saturated brine solution, dried over Na 2

SO

4 and evaporated. Purification was achieved by silica gel chromatography. Yield: 2.68 g (95 Preparation of B-1-vii: B-1-vi (5.61 mmol) was dissolved in aqueous TFA (90 15 ml) and further stirred at 0°C until completion. The reaction mixture was neutralized with aqueous NaOH solution at 0°C, concentrated in vacuo and dried. The residue was suspended in 90 ml acetylation mixture (pyridine/acetic anhydride 2/1) and 50 ml dichloromethane at 0°C and further stirred until completion. After concentration in vacuo and co evaporation with toluene, the residue was dissolved in ethyl acetate/H 2 0 the organic layer collected and washed 00 with 10 aqueous citric acid solution, saturated aqueous NaHCO 3 solution 0 and brine solution, dried over Na 2

SO

4 and evaporated. The crude residue and 3 p-thiocresol (6.0 mmol) were dissolved in 40 ml anhydrous dichloromethane L and cooled to 0°C, reacted with BF 3 xOEt 2 (8.41 mmol) and further stirred at rt until completion. The reaction was stopped with saturated NaHCO 3 solution and the organic layer was washed with water, dried over Na 2

SO

4 and evaporated. Final purification was achieved by silica gel chromatography to l n yield B-1-vii in 73 over 3 steps.

O

00 10 Preparation of B-1-xi: O B-1-x (8.0 mmol) was dissolved in 80 aqueous AcOH and heated at 100°C until completion. The mixture was cooled to rt, neutralized with solid NaHCO 3 and dissolved in ethyl acetate/water After removal of the aqueous layer, the organic layer was dried over MgSO 4 and evaporated to dryness furnishing B-1-xi in 98 yield.

Compound B-1: 'H-NMR (400 MHz, CDC13): 6= 8.07 2H, Aryl), 7.57-7.30 10H, Aryl), 7.10 2 H, Aryl), 6.88-6.81 4H, Mp), 5.55 1H, J1, 2 1.5 Hz,H-1p), 5.45 1H, 5.26 (ddd, 1H, 5.13 1H, 4.91 1H, Jgem 12.1 Hz, OCH 2 4.78 1H, Jgem 12.1 Hz, OCH 2 4.16 (dd, 1H, Jgem 9.6 Hz, J 5 6 a 7.6 Hz, H-6a), 4.08 (dd, 1H, J5,6b 5.2 Hz, H-6b), 3.93 1H, H- 3.77 3H, OCH 3 2.58-2.36 4H, (CH 2 2 Lev), 2.32 3H, SCH 3 2.05 3H, CH 3

C=O).

Example 4: Synthesis of L-ido configured qlvcosyl donor B-2.

HO OBn Ses HO OH 'SCres

C

OMpM OBn ~0 Sre OH OBz OMpm OBn -0 Sres B-2 LevO OBz Example 4: Synthesis of L-ido configured glycosyl donor B-2; a) SOP 2; b) SOP 18; c) SOP 5; d) SOP 23.

Examr~Ie 5: Svnthesis of Buildina Block C-I1, C-l a and C-l b g C-la-ill: R=SMe C-I b-ill: R=SCres C-l a: R=SMe C-I b: R=SCres C-Ia-il: R=SMe C-I b-li: R=SCres

OH

HO-

N

3 C-I-i-a: R=SMe C-I-i-b: R=SCres OAc ACO~- AcOV-'mm..OH C-I-ill

N

3

OH

HO-~ e H07 OTBDPS

N

3 C-1I-V OAc a AcO-~O

N

3 C-I-il-a: R=SMe C-I-il-b: R=SCres b OAc C AcO OT dP

N

3 C-I -iv 0 0 f IleOC

HOOBP

C-1 -vi3 Meo) z C-1-vii

N

3 OMpm Az

OTBDPS

C-1

N

3 00 Example 5: Synthesis of Building Blocks C-1, C-la and C-lb, conditions: a) O SOP 19; b) SOP 13, (78 2 steps); c) SOP 8, (91 d) SOP 22; e) SOP 2, 2 steps for C-l-vi f) SOP 18; g) SOP 5, (75 2 steps for C-1).

Preparation of C-1-iia: To methyl 2-azido-2-deoxy-1-thio-p-D-glucopyranoside. (10g, 42.50 mmol) in pyridine (50ml) at O°C was added acetic anhydride (20g) and the reaction stirred for 1 hour. The reaction mixture was evaporated to dryness and the q n residue extracted to give the title triacetate (15.23g, quantitative,), Rf=0.7 (CHCI3/Petroleum ethers, 1 0 o 10 Preparation of C-1-iii: To a solution of methyl 3,4,6-tri-O-acetyl-2-azido-2-deoxy-1-thio-p-Dglucopyranoside (14.1g, 39 mmol) in wet acetone (200ml) was added NBS (3 equiv.). The resulting mixture was allowed to stir for 2h. The mixture was then quenched, concentrated and the residue purified by silica gel chromatography to give the title hemiacetal as an oil (10.1g, Rf 0.5 (EtOAc/Petroleum ether, 1:1).

Preparation of C-1-vi: A mixture of 2-azido-2-deoxy-p-D-glucopyranosyl tert-butyldiphenylsilane (5.5g,12.42 mmol), 4-methoxybenzaldehyde dimethylacetal (4.4g, 24mmol), and 4-toluenesulphonic acid (100mg in acetonitrile/DMF (200ml, 5:3) were heated at 600C for 1 hour. The reaction mixture was then neutralized and evaporated to give the crude compound as an oil. The residue was purified by silica chromatography to give the product (6.7g, 96%, 85% from C-1-iv); Rf 0.8 (dichloromethane/Petroleum ethers; 10:2).

Preparation of C-1-vii: A mixture of DMAP (1.63g, 13.2 mmol) and benzoyl chloride (1.7g, 12.1mmol) and 2-azido-2-deoxy-4,6-O-(4-methoxybenzylidene)-p-D-glucopyranosyl tertbutyldiphenylsilane (6.7g, 11.9 mmol) in 1,2-dichloroethane (100mL) was stirred at 600C for 1h. The reaction mixture was quenched, extracted, washed and concentrated to give a crude residue. The residue was passed through a plug of silica to give the product (5.5g, Rf 0.7 (dichloromethane/Petroleum ethers; 4:1).

Preparation of C-1: 00 To a mixture of 2-azido-2-deoxy-3-O-benzoyl-4,6-O-(4-methoxybenzylidene)- 1-D-glucopyranosyl tert-butyldiphenylsilane (10Og, 1 sod i~mcyanoborohyd ride (5g, 75.6 mmol) and molecular sieves in DMF (200mnL) at 000 was added trifluoroacetic acid (28g, 247mmo1) at 000 and then left to run overnight at rt. The reaction mixture was quenched, filtered and concentrated and the residue purified by column chromatography to give the title compound (7.0g, 70% Rf 0.4 (ethylacetate/petroleumn ethers, 3:7).

Compound C-I: 001 S10 'H-NMR (400 MHz, CDCI 3 8.08 2 H, Aryl), 7.72 (in, 4 H, Aryl), 7.59 (in, c-i 1 H, Aryl), 7.47 (mn, 3 H, Aryl), 7.42 (in, 2 H, Aryl), 7.34 (in, 3 H, Aryl), 7.13 (d, 2 H, Mpm) 6.83 2H, Mpm), 4.96 (dd, I H, J 2 3 LJ34= 9.7 Hz, 4.53 (d, I H, J 1 2 7.6 Hz, H-I P3), 4.37 (2d, 2 H, OCH 2 3.83 (ddd, I H, 3.79 3 H, OCH 3 3.65 (dd, 1 H, 3.53 (dd, 1 H, Jgem 10.8 Hz, J 5 6 a 4.1 Hz, H- 6a), 3.46 (dd, 1 H, J5,6b 4.1 Hz, H-6b), 3.12 (in, 1 H, 3.02 1 H, J 4

,OH

3.5 Hz, 4-OH), 1.12 9 H, C(CH 3 3 Example 6: Synthesis of Building Block C-2 HO 0 a 0 b H O07 R R

N

3

N

3 C-i-i-a: R=SMe C-2-i-a: R=SMe C-I-i-b: R=SCres C-2-i-b: R=SCres

OH

C HO d BOR 0 BzO R 00R

N

3

N

3 C-2-ii-a: R=SMe C-2-iii-a: R=SMe C-2-ii-b: R=SCres C-2-iii-b: R=SCres OBz OBz Of z Rz e A OH HO OTBDPS

N

3 C-2- v N 3 C-2 N 3 C-2-iv-a: R=SMe C-2-iv-b: R=SCres 00 Examnple 6: Synthesis of Building Block C-2, conditions: a) SOP 1, (90% for R=SMe); b) SOP 18, (87% for R=SMe); c) SOP 4, p-TosOH, MeOH, CH 3

CN

(86% for R=SMe); d) SOP 17, (92% for R=SMe); e) SOP 13, f) SOP 8, Compound C-2: 1 H-NMR (400 MHz, CDC1 3 5= 8.09 2 H, Aryl), 7.97 2 H, Aryl), 7.72 (in, 4 H, Aryl), 7.60 (in, 1 H, Aryl), 7.50-7.27 (in, 11 H, Aryl), 4.98 (dd, 1 H, J 2 3 J34= 9.7 Hz, 4.58 1 H, J1, 2 7.8 Hz, H-I P3), 4.51 (dd, 1 H, Jgem 11.3 Hz, J 5 6 a 4.7 Hz, H-6a), 4.36 (dd, 1 H, J 5 6 b 2.2 Hz, H-6b), 3.72-3.68 (in, 2 00 H, H-2, 3.31 (in, 1 H, 3.23 1 H, J 4 ,OH 4.5 Hz, 4-OH), 1. 13 9 H, C(CH 3 3 Example 7: Synthesis of several carbamoytated Building Blocks C-3a to C-3d and C-4a to C-4d, containing a 6-0 benzoyl or protection

OH

HO 01 RHO R 0 R R HMeO-3i N 1 HO~ 3 H -0 ~O pm- Z C-3a: R I SCres C-3b: R 1 =SEt C-3c: R 1

OTBDPS

C-3d: R1 SMe C-4a: R' SCres C-4b: R' SEt C-4c: R' OTBDPS C-4d: R =SMe 00 Example 7: Synthesis of several carbamnoylated building blocks C-3a to C-3d and C-4a to C-4d, containing a 6-0-benzoyl or protection, conditions: a) R =MeO: SOP 2; R H: SOP 1, (82 %,R 1 =SCres, 5 R b) SOP 30, (87 %,R 1 =S~res, R c) SOP 29, (95 %,R1 Scres, R d) SOP 4, (72 %,R 1 =S~res); e) SOP 17, (85 f) SOP Compound C-3a: 1 H-NMR (400 MHz, 0D01 3 5= 8.06 2 H, Aryl), 7.62 (in, 1 H, Aryl), 7.48 (t, 00 10 2 H, Aryl), 7.38 2 H, Aryl), 6.97 2 H, Aryl), 5.06 (bs, 1IH, NH), 4.79 (dd, 1IH, Jgem 12.0 Hz, J 5 6 a, 3.6 Hz, H-6a), 4.70 1IH, J 1 2 9.2 Hz, H-1 4.63 (dd, 1 H, J5,6b 2.0 Hz, H-6b), 4.18 (dd, 1 H, J 2 3 J34= 10.4 Hz, 3.89 (dd, 1IH, J 4 5 9.2 Hz, 3.72 (in, 1IH, 3.23 (ddd, 1IH, 3.12 (bs, 1 H, 4-OH), 2.29 3H, S0H 3 Examp~le 8: Synthesis of several 6-OMD and cyclic 2,3-carbamoyl protected buildinci blocks C-5a to 00 ou P 0i NPht Pht HO' R c HO d SBzO RBzO R1 O C-5-ii Pht C-5-iii Pht HO e HO-\ NH2 R SMe

R

1 SEt

R

1

OTBDPS

Example 8: Synthesis of several 6-OMp and cyclic 2,3-carbamoyl protected building blocks C-5a to C-5c, conditions: a) SOP 18, (92 for R' OTBDPS); b) SOP 4, (82 c) SOP 26, (75 for R 1 OTBDPS); d) SOP 30, (87 e) SOP 29, (95 Compound 1 H-NMR (400 MHz, CDCI 3 6= 7.69 2 H, Aryl), 7.63 2H, Aryl), 7.46- 7.31 6H, Aryl), 6.82 (bs, 4H, Mp), 5.04 bs, 1H, NH), 4.78 1H, J 1 2 7.6 Hz, H-1p), 4.15-4.10 3H, H's not assigned), 3.97 (dd, 1H, J =11.6 Hz, J 9.6 Hz, H not assigned), 3.78 3H, OMe), 3.56 1H, H not assigned), 3.48 1H, H not assigned), 2.80 (bs, 1 H, 4-OH), 1.08 9H, C-(CH 3 3 Example 9: Synthesis of Building Blocks C-6-a and C-6-b and C-7 MeO 0 00MeC

NH

2 C-3-iii-a: R=SCres C-3-iii-f: R=SMe MeO IJA C0Mpm R 0

N

3 C-6-ii-a: R=SCres C-6-ii-b: R=SMe b "3 C-6-i-a: R=SCres C-6-i-b: R=SMe Mpmo pm Mpm O R

N

3 C-6-a: R=SCres C-6-b: R=SMe OMpm MpmO OH C-74i N 3 e OMpm HO 0 MpmOv OTBDPS C-7 N 3 Example 9: Synthesis of building blocks C-6-a, C-6-b and C-7, conditions; a) SOP 12, (83 b) SOP 7; c) SOP 5, (75 2steps); d) SOP 14 82 e) SOP 8 (91 Compound 0-6-a: 1 H-NMR (400 MHz, CDCI 3 5= 7.43 2 H, Aryl), 7.25 (in, 4 H, Aryl), 7.08 (d, 2 H, Aryl), 6.88 (in, 4 H, Aryl), 4.81 1 H, Jge m 10.8 Hz, OCH 2 4.74 1 H, Jgem 10.8 Hz, OCH 2 4.53 1 H, Jgem =11. 1 Hz, OCH 2 4.48 1 H, Jgern 10.8 Hz, OCH 2 4.35 1 H, J 12 =10.0 Hz, H-1p), 3.82 3 H,

OCH

3 3.79 3 H, OCH 3 3.76 (dd, 1 H, Jgem 10.5 Hz, J 5 5.4 Hz, H- 6a), 3.70 (dd, 1 H, J5s6b 5.4 Hz, H-6b), 3.56 (ddd, 1 H, 3.42 (in, 1 H, H- 3.34 (dd, 1 H, J 3 4 =8.8 Hz, 3.24 (dd, 1 H, J 23 9.4 Hz, 2.72 1 H, J 4 .OH 3.5 Hz, 4-OH), 2.38 3 H, SCH 3 Comp~ound C-7: 'H-NMR (400 MHz, CDC1 3 7.63 4 H, Aryl), 7.35-7.21 (mn, 8H, Aryl), 7.08 (mn, 2 H, Aryl), 6.83-6.78 (mn, 4 H, Aryl), 4.72 1 H, Jgem 11.0 Hz, OCH 2 4.59 1 H, Jgem 11.0 Hz, OCH 2 4.29 1 H, J 1 2 7.8 Hz, H-1 4.27 (d, 1H, Jgem 11.7 Hz, OCH 2 4.21 1H, Jgem 11.7 Hz, OCH 2 3.72 3 H, OCHA) 3.71 3 H, OCH 3 3.51 (ddd, 1 H, J 3 4 ZJ45= 8.6 Hz, 3.40- 3.32 (in, 3 H, H-6a, H-6b, 3.05 (dd, 1 H, J 2 3 9.8 Hz, 2.90 (in, 1 H, 2.51 1 H, J 4 .OH 2.2 Hz, 4-OH), 1. 12 9 H, C(CH 3 3 Example 10: Synthesis of building block C-8a to C-Bc HO R C-3-ii-c NPht

OH

HO 0 A110aR C-841i NPht OMp HO 0 A1II-% R C-B-iv NH 2 a-,1 1 0 C-B-i NPht OMp C HO 0 C-B-lj NPht ONIp

N

3 C-Ba R SIVe C-8b R SlEt C-Bc R OTBDPS b d Example 10: Synthesis of building blocks C-8a to C-Bc, conditions: a) SOP 7, AIIBr, DMF (65 R=OTBDPS); b) SOP 4, (86 R=OTBDPS); c) SOP 26, R=OTBDPS); d) SOP-30; e) SOP 12, (70 2 steps for R=OTBDPS).

Compound C-Bc: 1 H-NMR (400 MHz, 00013): 5= 7.72 (in, 4 H, Aryl), 7.43-7.16 (in, 6 H, Aryl), 6.76 (in, 4H, Mp), 5.96 (in, 1 H, =CH Allyl), 5.31 (in, 1 H, =CH Allyl), 5.22 (in, 1 H, =CH Allyl), 4.42 1 H, J 1 2 7.6 Hz, H-1 4.39 (in, 1 H, OCH 2 Allyl), 4.23 (in, 1 H, OCH 2 Allyl), 3.97 (dd, 1 H, Jgem =10.0 Hz, J 5 6 a 3.6 Hz, H-6a), 3.92 (dd, 1 H, J 5 6 b 5.2 Hz, H-6b), 3.77 3 H, 00H 3 3.66 (ddd, 1 H, J 4 5

J

3 4 9.4 Hz, 3.42 dd, 1 H, J 9.8 Hz and J 7.8 Hz, H not assigned), 00 3.22 1 H, 3.09 (dd, 1 H, J 8.4 Hz and J 9.6 Hz, H not assigned),

O

S2.48 1 H, J 4 ,OH 2.8 Hz, 4-OH), 1.12 9 H, C(CH 3 3 T Example 11: Synthesis of Building Block D-1: a b [HO- SMe P h SMe_ 0 o O SMe a o5 b OH D-1-i OH 00 h 0o cPh o BO H SOBn DBnO i SMe Bn1 OH D-1-ii OBn D-1-iii OBn

OHO

Ph' e HO f no OTBDPS ON nO TBDPS 0 D-1-iv OBn D-l-v OBn BnOY

OTBDPS

D-1 OBn Example 11: Synthesis of building block D-1; a) SOP 1, (95 b) SOP 7, c) SOP 13, (92 d) SOP 8; e) SOP 4, (70 2 steps); f) 1. SOP 15; 2.

SOP 16, (75 2 steps).

Compound D-1: 1 H-NMR (400 MHz, CDCI 3 S= 7.72 4 H, Aryl), 7.41 2 H, Aryl), 7.32 7.25 14 H, Aryl), 5.04 1 H, Jgem 11.0 Hz, OCH 2 4.81 3 H,

OCH

2 4.63 1 H, J 1 2 7.4 Hz, H-1 3.88 (ddd, 1 H, J3, 4 A J4,5 9.2 Hz, H- 3.70 3 H, OCH 3 3.53 (dd, 1H, J 7.5 Hz, J 9.0 Hz, H not assigned), 3.47 1 H, J 4 5 9.8 Hz, 3.42 (dd, 1 H, J 8.9 Hz and J 8.9 Hz, H not assigned), 2.87 1 H, J 4 ,oH 2.4 Hz, 4-OH), 1.11 9 H, C(CH 3 3 00 Exanple 12: Synthesis of Building Block D-2, a 2-O-Allvloxycarbonyl protected thioethyl gilycoside QAc INDAcO:0Ph' 0 AC a 00b C) AcO O AcHO 00 D-24

H

D D-2-ii PO OTrit 0 C LevO No- a O BnO BnO 0-2-iii D-2-iv 0 OMe LevO 0 e Le BnO BnO SEt OAIloc D-2-v D-2 Example 12: Synthesis of Building Block D-2, conditions: a) 1. SOP 22; 2.

SOP 1; b) SOP 7; c) 1. SOP 4; 2. TritCl, Pyridine, (CICH 2 2 3. SOP 23; d) 1.

Cr0 3

H

2 S0 4 Acetone, 00C, 2. SOP 16; e) 1. Dimethyl dioxirane Acetone; 2.

EtSH, TFAA, CH 2

CI

2 3. SOP Example 13: Synthesis of Building Block D-3 B-I -ii OBz a 0z~-9 b BnO m.OBz OBz D-3-i C HO~ d PS N BnO OTBDPS BzO D-3-Iiii OBz Bz Zo-- OTBDI OBz D-3-ii o OMe Bno! OTBI BzO D-3-iv e DPS 0 OMe BnO ,a~OTBDPS D-3-v BzO 0 OMe LevO -0 gO, BnO

OH

BzO D-3-vi 0 We D-3 BzO Example 13: Synthesis of Building Block D-3, conditions: a) 1. SOP 4, Amberlite IR 120, H 2 0, 8000; 2. SOP 18, (85 2 steps) b) 1. SOP 21; 2.

SOP 8, 70 2 steps); c) SOP 22, (96 d) 1. SOP 15; 2. SOP 16, (80 2 steps); e) SOP 23, (92 f) SOP 9, (95 g) SOP 25a, (91%) Preparation of D-34i, step 1: The starting material (57 mmol) and Amberlite IR 120 iron exchange resin (Hfform, 20 g) were suspended in water (180 ml) and stirred at 80 00 until completion. The iron exchange resin was removed by filtration and extracted with water The combined aqueous layers were neutralized with triethyl amine and freeze dried.

Compound D-3-v:.

00 'H-NMR (400 MHz, CDCI 3 9= 7.95 (in, 2 H, Aryl), 7.68 (in, 2 H, Aryl), 7.58 7.12 (in, 16 H, Aryl), 5.47 (dd, J 1 2 7.6 Hz, J 2 3 9.6 Hz, 5.31 (dd, J 4 4 9.6 Hz, 4.64 1 H, 7.6 Hz, H-1 4.60 1 H, Jgem, 12.0 Hz, OCHA) 4.55 1IH, Jgem 12.0 Hz, OCH 2 3.74 (dd, 1 H, 3.70 3H, OCHA) S 5 3.63 1IH, J 4 5 9.6 Hz, 2.68 -2.16 (mn, 4H, (0H 2 2 -Lev), 2.15 3H,

CH

3 0.96 9H, C(CH 3 3 Example 14: Syntheses of a range of block D donor sugars D-4 to D-7 from a common intermediate, a 4-0-levulinoyl glucal.

000 LevO 0 0 OMe D6:RCc D-6a: R=AIlocO D-4 OAIIoc 0 OMe 0 OMe LevO Lev0- 0 Bn~ BnO (S0)Ar CIAcO D-2-v D-7 BnO O(CH 2 3

CH=CH

2 OAIIoc Example 14: Syntheses of D-4 to D-7 as donor sugars, conditions: a) 1.

Dimethyl dioxirane, Acetone; 2. TBAF, THE; 3. SOP 35; b) 1. Dimethyl dioxirane, Acetone; 2. 4-penten-1-oI, ZnCI 2 0H 2 01 2 3. SOP 35; c) 1. Dimethyl dioxirane, Acetone; 2. ArSH, TFAA, CH 2

CI

2 (Ar Ph, p-Tol); 3. SOP 35 or (CIAc) 2 0, Pyridine, CH 2 0 2 (for D-6b); d) MCPBA, 0H 2 C1 2 (for D-6b as substrate).

2008200567 07 Feb 2008 Example 15: Synthesis of Building Block E-1 to E-4

OTBDPS

BnO 0~ B nO m E-4

N

3 1

OTCA

OTBDPS

i- BnO% R

N

3 E-I-ii-a: R=SMe E-I -li-b: R=SCres

OH

HO~ R

N

3 C-I-i-a: R=SMe C-I-i-b: R=SCres E-I-iii-a: R=SMe N 3 E-I-iii-b: R=SCres

OTBDPS

N

3 E-I-i-a: R=SMe E-l-i-b: R=SCres OBz d BnO7 R E-I-iv-a: R=SMe N 3 E-1-iv-b: R=SCres OMp

N

3 E-I-v-a: R=SMe E-I-v-b: R=SCres OMpm BnO0 Bn&~

N

3 E-I-vi-a: R=SMe E-l-vi-b: R=SCres \OBz BnO 0~ BnOAm.

OTCA

e$

"OMP

OMpm BnO~- BnO t 10E-2 N1

OTCA

BnO.

OTCA

Example 15: Synthesis of Building Block E-1 to E-4, conditions: a) SOP 8; b) SOP 7; c) SOP 9, (84% over 3 steps, R=SMe); d) SOP 18, R=SMe); e) 1. SOP 13, for E-1 -iv-a as starting material); 2. SOP 25b, f) 1.

Tos~l, Pyridine; 2. p-MeO-C 6

H

4 -ONa, NMP, 6000; g) SOP 7, (78 R=SMe); h) 1. SOP 14; 2. SOP 25b, (79 2 steps, R=SMe).

Preparation of E-1-i-a: 00 A mixture of methyl 2-azido-2-deoxy-thio-p-D-glucopyranoside

O

S42.5mmol) and imidazole (4.9 g, 71.25 mmol) in 20 mL DMF was treated with tert-butyldiphenylchlorosilane (11.6mL, 44.63mmol) for 2h. The reaction Smixture was concentrated, extracted, washed and dried, yield: 23g (crude light yellow syrup), Rf 0.74 (CHCI3/methanol 9/1).

Preparation of E-1-ii-a: The silyl ether from the previous step in 50 mL DMF, was treated with 2.68g of g 95% NaH (106.25 mmol) and 12.64 mL (106.25 mmol) of benzyl bromide at 0 °0oC. After 1h the excess NaH was quenched and the reaction concentrated, extracted, washed and concentrated to afford a yellow syrup yield: 28.5g (crude yellow syrup), Rf 0.80 (hexane/ethyl acetate Preparation of E-1-iii-a: The crude yellow syrup from the above reaction was treated with 36.5 mL AcOH and 106.3 mL (106.25 mmol) of 1M solution of TBAF in THF overnight.

The reaction was concentrated and purified by chromatography to afford the title compound. 14.9g 3 steps Rf= 0.36 (petroleum spirit/ethyl acetate 7/3) Preparation of E-1-iv-a: Methyl 2-azido-2,3 di-O-benzyl-2-deoxy-thio-p-D-glucopyranoside (14.5 g, 34.9 mmol) in dichloromethane (200 ml) and anhydrous pyridine (8.6 ml, 106.2 mmol) was treated with benzoylchloride (4.93 ml, 42.5 mmol) at 0°C for 1 hour. The reaction mixture was quenched, extracted, washed and evaporated. The residue was purified by silica gel column chromatography to afford the title compound as a white solid.

Yield: 14.9 g (82 Rf 0.82 (Petroleum spirit/Ethyl acetate 7/3).

Preparation of E-1: Methyl 2-azido-6-O-benzoyl-2,3di-O-benzyl-2-deoxy-thio-C-Dglucopyranoside (8.68 g, 16.7 mmol) in acetone (50 ml) was treated with Nbromosuccinimide (8.92 g, 50.12 mmol) at 0°C for 1 hour. The reaction mixture was then quenched, extracted, washed and evaporated, furnishing a yellow syrup which was purified by chromatography. Yield: 6.13 g Rf 0.57 (Petroleum spirit/Ethyl acetate A cooled mixture of 2-azido-6-O- 00 benzoyl-2,3 di-O-benzyl- 2-deoxy-ac/3-D-glIucopyra nose (5g, 10.2 mmol),

K

2 00 3 (7.0 g 51 mmol) and trichioroacetonitrile (5.1lmI, 51 mmol) in 30 ml of dichioromethane was stirred for 2h. The mixture was then filtered through celite and the filtrate was concentrated and purified on a short column of silica S 5 gel to obtain the title compound as an amorphous white solid. Yield 5.69 g Rf 0.85 (Petroleum spirit/Ethyl acetate 7/3).

Compound E-1: 1 H-NMR (400 MHz, CDCI 3 5= 8.73 1 H, C=NH), 8.00 (in, 2 H, Aryl), 7.56 00 C) 10 (in, 1 H, Aryl), 7.43-7.25 (in, 12 H, Aryl), 5.66 1 H, J 8.4 Hz, H-1 4.95 (1 (d,I1H, Jgm 10.8 Hz, OCHA) 4.87 2H, J 10.8 Hz OCHA) 4.62 2H, Jgem 10.8 Hz, OCH 2 4.58 (dd, 1 H, Jgem= 12.4 Hz, J 56 a 2.0 Hz, H-6a), 4.46 (dd, 1IH, J5,6b 3.6 Hz, H-6b), 3.77-3.72 (mn, 3H, H-5, 2H not assigned), 3.62 (dd, 1 H, J 8.3 Hz, J 9.7 Hz, H not assigned).

Comp~ound E-2: 1 H-NMR (400 MHz, CDCI 3 1 H, C=NH), 7.38-7.22 (in, 10H, Aryl), 7.13 (mn, 2H, Aryl), 6.83 2H, Mpm), 6.44 1IH, J 12 3.5 Hz, H-1lc), 4.93 1H, Jgem 10.5 Hz, OCH 2 4.89 1H, Jgem 10.5 Hz, OCH 2 4.78 (d, 1H, Jgem 10.5 Hz, OCH 2 4.57 1H, Jgem 11.7 Hz, OCH 2 4.51 1H, Jgem 11.7 Hz, OCH 2 4.39 1H, Jgem 11.7 Hz, OCH 2 4.02 (dd, 1H, J 3 4 ;J23= 9.5 Hz, 3.98 (in, 1 H, 3.86 (dd, 1 H, J 45 9.6 Hz, 3.76 (dd, 1 H, 3.75 3H, OCH 3 3.69 (dd, 1IH, J 5 ,6a 3.5 Hz, Jgem 10.5 Hz, H-6a), 3.63 (dd, 1 H, J5.6b 1.8 Hz, H-6b).

Example 16: Synthesis of Building Blocks E-5 to E-8 P h-40e 3 V Bn o SR E-1-vii-a: R=Cres NPht E-1-vii-b: R=Me HO a BnO SR NPht E-1-viii-a: R=Cres E-1-viii-b: R=Me

TBDPSO

DTPMNH

OTCA

E-81

TBDPSO

Bno SR

DTPMNH

E-8-i-a: R=Cres E-8-i-b: R=Me Bno

SR

DTPMNH

R=Cres R=Me NMe 2 0 .SR DTPM-NMe 2

C

NH

2 E-1-ix-a: R=Cres E-1-ix-b: R=Me

MPMO-

BnO SR

DTPMNH

E-6-i-a: R=Cres E-6-i-b: R=Me 9pO

IMPO-

BnO 0~- BnO -mmA.SR

DTPMNH

E-7.i-a: R=Cres E-7-i-b: R=Me MpO BnO\ 0 BnOamm

DTPMNHI

E-7 OTCA \d BzO BnO 0~ BnO~~m4 SR

DTPMNH

R=Cres E-5-ii-b: R=Me BzO BnO BnO IDTtPMNHI E-5 OTCA Example 16: Syntheses of Building Blocks E-5 to E-8, conditions: a) SOP 6, (85 R=SMe); b) SOP 30, (86 R=SMe); c) SOP 10, (88 R=SMe); d) SOP 18, (92 R=SMe); e) 1. SOP 13; 2. SOP 25b, (85 2 steps, R=SMe); f) SOP 7; g) 1. SOP 14; 2. SOP 25b; h) 1. TsCI, DMF; 2. p-MeO-0 6

H

4 -ONa, NMP, 60'0; i) SOP 8.

00 Compound 'H-NMR (400 MHz, CDCI 3 5= 10.20 (dd, 1 H, JNH,=C-H 14.0 Hz, JNH,H-2= 9.9 Hz, NH), 8.80 1 H, C=NH), 8.16 1H, 7.99 (in, 2H, Aryl) 7.58 (in, 1 H, Aryl), 7.45 (in, 2 H, Aryl), 7.30-7.17 (in, 1 OH, Aryl), 6.42 1 H, J1, 2 S 5 3.6 Hz, H-lac), 4.89 1H, Jge.m =8.4 Hz, 00HA) 4.68-4.60 (in, 3H, OCH 2 4.58 (dd, 1IH, J 5 6 2.0 Hz, Jgem =12.4 Hz, H-6a), 4.51 (dd, 1 H, J5,6b Hz, H-6b), 4.22 (in, 1 H, 4.03 (dd, 1 H, J 3 4 'z 23= 9.6 Hz, 3.80 (dd, 1 H, J 4 5 9.4 Hz, 3.70 (ddd, 1 H, 3.32 3 H, NCH 3 3.25 c-i 3 H, NCH 3 00 Example 17: Preparation of L-iduronic acid containing disaccharides B-A-I to B-A- 1 es B-1I B-4

R

2 Mp Mpm B-A-1: R 1 N3 R 2 Mp B-A-2: R 1

N

3 R Mpm B-A-3: R 1 DTPMNH, R 2 Mp B-A-4: R 1 DTPMNH, R 2 Mpm L<

I-

R

1

N

3 B-A-6: R 1

DTPMNH

L

B-A-7: R 1

N

3 B-A-8: R 1

DTPMNH

B-A-9: R 1

N

3 R DTPMNH OBz HO 0 BnO-

R

OMe SA-1: R N 3 A-3: R 1

DTPMNH

SOBz OR OBn 0 BnO O On R 2:L OMe svO OBz b V .OBz 1H OBn 0 -Bn 0 R I OMe evO OBz SOBz MeO OBn OBO 0 R OMe LevO OBz d OBz 0 MeO OBn 0 BnO O R' O OMe HO OBz Example 17: Preparation L-iduronic acid containing disaccharides B-A-1 to Ba) SOP 32a, (76 for b) SOP 27, (88 for c) 1.SOP 20; 2. SOP 16, (84 for B-A-7, 2 steps); d) SOP 24, (94 for B-A-9).

Formation of disaccharide B-A-1 (step a) A suspension of A-1 (410 mg, 992 pmol), B-1 (680 mg, 992 pmol) and freshly activated molecular sieves 4 A (1.0 g) in dry CH 2

C

2 (20 mL) was stirred for min at 0°C. N-lodosuccinimide (405 mg, 1.8 mmol) was added and stirring 00 continued for 20 min. After addition of trifluoromethanesulfonic acid (10.6 pl,

O

0 119.7 pmol), the reaction mixture was further stirred until completion (from S0°C to 25 and quenched with aqueous NaHCO 3 -solution (10 The C. mixture was diluted with CH 2 01 2 and filtered through a celite pad. The filtrate was washed with a 10 KHCO3/Na 2

S

2 0 3 solution, water and saturated brine solution, dried over MgSO 4 and evaporated. Final purification was achieved by silica gel column chromatography. Yield: 730 mg (76 SFormation of disaccharide B-A-7(step c) 00 10 Disaccharide B-A-5 (1.00 g, 1.15 mol) was dissolved in anhydrous DMF Sml) and reacted with pyridinium dichromate (4.33 g, 11.5 mmol) under stirring at room temperature until complete conversion into the uronic acid. The reaction mixture was subsequently poured into 50 ml water and the whole extracted with diethyl ether. The combined ether layers were washed with aqueous citric acid solution, filtered through a short silica gel pad, dried over MgSO 4 evaporated and dried under high vacuum. The crude residue was dissolved in Toluene (3 ml) and methanol (3 ml) and titurated with

TMSCHN

2 solution (2M in hexane) until completion. The excess of TMSCHN 2 was destroyed by addition of acetic acid and the mixture evaporated. Final purification was achieved via silica gel chromatography. Yield: 871 mg (84 Compound B-A-9: 1 H-NMR (400 MHz, CDC1 3 8= 8.03 2H, Aryl), 7.91 2H, Aryl), 7.53 2H, Aryl), 7.42-7.23 14H, Aryl), 5.37 1H, J1, 2 1.5 Hz, 5.21 1H, 4.97 1H, J 4 5 2.3 Hz, 4.84 2H, Jgem 10.8 Hz,

OCH

2 4.81 1H, Jgem 10.8 Hz, OCH 2 4.80 1H, J1, 2 3.6 Hz, H-la), 4.77 (1H, J 5 6 a 1.8 Hz, H-6a), 4.70 2H, OCH 2 4.47 (dd, 1H, J 5 6 b 4.2 Hz, Jgem 12.3 Hz, H-6b), 4.05-3.97 3H, H-4, 3.91-3.87 (m, 2H, 3.49 3H, OCH 3 3.44 1H, 3.43 3H, OCH 3 Selected 13 C-NMR (100 MHz, CDCI 3 8= 98.73 C-1 (JcH 172.5 Hz), 98.35 C-1' (JcH 171.8 Hz).

00 Example 18: Syntheses of Buildingj Blocks E-D-11 to E-D-12

OR

1

OR

1 n0 R00 BnO OBP E-1, E-2, E-3, E-5, E-6, E-7, U~ E-1-v-b, E-1-vi-a, E-1-vi-b 1n E-1-a, E-1-b, E-5-ii-a, E-5-ii-b E-D-1 R =N 3 R Bz E-6-i-a, E-6-i-b, E-7-i-a, E-74-b E-D-2: R =N 3

R

1 Mpm E-D-3: R N 3 R MP aN E-D-4: RDTPMNH, R B 00 o Oe E-D-5: R =DTPMNH, R I Mpm HO E-D-6: R=DTPMNH, R =MWpm OBn D-1 O I b BnO0 Bn& o OMe E-D-7: R N 3 RI Bz 0- E-D-8: R N 3

R

1 =P Mpm

OTCA

E-D-9: R N 3

R

1 Mp E-D-1O: R DTPMNH, R 1 Bz E-D-1 1 :R DTPMNH, R 1 Mpm E-D-12: R DTPMNH, R 1 Mpm Example 18 :Syntheses of disaccharides E-D-1 to E-D-12, conditions: a) SOP 32 alb for X =SMe/ Scres or SOP 33 for X OTCA, (88 for E-D-1 via E-1, 84% for E-D-4 via E-5, as 4~ mixtures), b) 1. SOP 9; 2. SOP 25a, (90 for E-D-7 over 2 steps).

Preparation of E-D-1 :Methyl (2-azido-6-O-benzoyl-3,4-di-O-benzyl-2-deoxy-aX- 0-gI ucopyranosyl)-(l1-*.4)-tert-butyld iphenylsilyI 2 ,3-d i-O-benzyl-f3-Dglucopyranosid)uronate A mixture of 2-azido-6-O-benzoyl-2,3 di-O-benzyl- 2-deoxy-a/3-Dglucopyranosyl trichloroacetimidate (2.5 g, 3.94 mmol), and methyl (tedtbutyldiphenylsilyl 2,3-di-O-benzyl-f3-D-glucopyranoside) uronate (1.6 g, 2.55 00 mmol) and molecular sieves 4A (2.5 g) in 50 ml diethyl ether was treated with TBDMSO~f (180 pi, 788.76 uimol) at -200C for 1lh. The reaction was quenched filtered, concentrated and the residue purified by silica gel column chromatography to obtain the desired disaccharide 2.48 g, 88%. Rf= 0.67 S 5 (toluene/ethyl acetate 9/1) Compound E-D-1: E-D-1 was formed according to SOP 33 with ether as solvent at -300C and TBDMSOTf as promotor in 86 yield (cc/3-mixture).

001 1H-NMR (400 MHz, CDCI 3 5= 8.00 (in, 2H, Aryl), 7.68 (in, 4H, Aryl), 7.56 (in, ci~ 1H, Aryl), 7.42 (in, 4H, Aryl), 7.36-7.17 (in, 24H, Aryl), 5.47 1 H, J 1 2 3.8 Hz, H-I 5.02 1 H, Jgem 11.4 Hz, OCH 2 4.97 1 H, Jgemll1.0 Hz, OCHA) 4.84 (in, 4H, OCH 2 4.75 1 H, Jgem 11.4 Hz, 00HA) 4.66 1 H, J12= 7.5 Hz, H-1 4.57 1IH, Jgem 10.9 Hz, OCH 2 4.45 (in, 2H, H-6'a, 4.15 (dd, J 8.8 Hz and J 9.6 Hz), 3.86 (in, 1 3.65 3H, 00H 3 3.68-3.58 (in, 3H), 3.55 1 H, J 4 5 10.0 Hz, 3.31 (dd, 1 H, J 2 3 10.2 Hz, 1. 12 9H, C(CH 3 3 Compound E-D-4: E-D-4 was formed according to SOP 33 with ether as solvent at -300C and TBDMSOTf as promotor in 84 yield (a/p-iixture).

Selected 1 H-NMR (400 MHz, 00013): 6= 10.02 (dd, 1 H, JNH,=c-H 14.4 Hz, JNH,H-2 9.6 Hz, 8.02 (in, 2 H, Aryl), 7.79 1 H, 7.72-6.93 (mn, 33 H, Aryl), 5.60 1 H, J1, 2 3.6 Hz, 4.49 1 H, J 1 2 7.8 HZ, H-1 P), 3.66 3 H, OCH 3 3.29 3 H, NCH 3 3.28 3 H, NCH 3 1.14 9 H, C(CH3)3).

Preparation of E-D-7: Methyl (2-azido-6-O-benzoyl-3, 4-di-O-benzyl-2-deoxya-D-glUcopyranosyl)-(1 3-di-O-benZyl-fJ-D-glUcopyranosyl trichloroacetimidyl)uronate A solution of methyl (2-azido-6-O-benzoyl-3,4-d i-O-benzyl-2-deoxy-at-D-glUCOpyranosyl)-(l1-+4)-tert-butyldiphenylsilyI 2 ,3-d i-O-benzyl-j3-D-glucopyranoside) 00 uronate (2.09 g, 1.90 mmol) in acetic acid (1.74 ml, 30.45 mmol) and 1 M

O

Ssolution of tetrabutylammoniumfluoride (7.6 ml, 7.61 mmol) was stirred at room temperature overnight. The reaction mixture was then concentrated and Cthe residual syrup was purified by silica gel column chromatography to obtain the desired hemiacetal. Yield: 1.57 g Rf= 0.21 (toluene/ethyl acetate 9/1).

t- A mixture of methyl (2-azido-6-O-benzoyl-3,4-di-O-benzyl-2-deoxy-a-Dglucopyranosyl)-(1->4)-2,3-di-O-benzyl-p-D-glucopyranosyl)uronate (594 mg, C 690.70 [mol), trichloroacetonitrile (280 il 2.74 mmol) and DBU (31 pl, 209.3 00 0 10 p.mol) in 8.0 ml dichloromethane was stirred at 0°C for 1 h. The mixture was 0 N then concentrated and purified on a short column of silica gel to obtain the title compound as an amorphous white solid. Yield: 662 mg (95.3 Rf= 0.46 (toluene/ethyl acetate 9/1).

Compound E-D-7: Selected 1 H-NMR (400 MHz, CDCI 3 6= 8.68 1H, C=NH), 8.00 2H, Aryl), 7.56 2H, Aryl), 7.43-7.23 22H, Aryl), 6.48 1H, J 1 2 4.3 Hz, H-la), 5.59 1H, J 1 2 3.6 Hz, 5.03 (1H, Jgem 10.8 Hz, OCH 2 4.93-4.83 4H, OCH 2 4.70 1H, Jgem= 12.0 Hz, OCH 2 4.64 1H, Jgem 12.0 Hz, OCH 2 4.60 1H, Jgem= 11.2 Hz, OCH 2 4.47 2H, H- 6'a, 4.42 1H, not assigned), 4.15 2H, not assigned), 3.97 (dd, 1H, J 8.2 Hz and J 10.2 Hz, not assigned), 3.80 1H, not assigned), 3.76 3H, OCH 3 3.72-3.64 2H, not assigned), 3.30 (dd, 1H, J 23 10.4 Hz, Example 19: Syntheses of disaccharides E-D-13 to E-D-44 R Ac, Alloc, Bz, Piv E-D-13 to E-D-28

OR'

BnO- 0~O BnO-. ~X R2 E-1, E-2, E-5, E-6, E-1 -iv-a, E-1 -iv-b, E-1-vi-a, E-1-vi-b, E-5-ii-a, E-6-i-a, E-6-i-b

R

1 0 E-D-29 to E-D-44 E-D-1 3 E-D-29: R 3 =Bz, R 2

=N

3

R

1 Ailac E-D-14 E-D-30: R 3 Bz, R 2 NHDTPM, R 1 Alloc E-D-I 5 E-D-31 :R 3= Mpm, R 2= N 3 R1 Alloc E-D-1 6& E-D-32: R 3 =Mpm, R 2 =NHDTPM, R 1 AI1oc E-D-1 7& E-D-33: R 3 Bz, R 2

N

3

R

1 Piv E-D-I 8 E-D-34: R 3 =Bz, R 2 =NHDTPM, R1 Piv E-D-19 E-D-35: R Mpm, R 2

=N

3

R

1 Piv E-D-36: R 3 Mpm, R 2 NHDTPM, R1 Piy E-D-21 E-D-37: R 3= Bz, R 2= N 3 R1 Bz E-D22& -D38:R3 z 2 =IHTM,'B E-D-23 E-D-38: R3 Bz, R N3DP, R Bz E-D-24 E-D-40: R 3 Mpm, R 2

N

3 D, R 1 Bz E-D-25 E-D-40: R 3 =Mp, R 2 NHDPM R 1 E-D-2 E-D-42: R 3= Bz, R 2=N 3 ,RPM R1Ac A E--7&ED4:R3 =M ,2 I3 1=A E-D-26 E-D-42: R 3B=p,R NHDTPM, R Ac Example 19: Syntheses of disaccharides E-D-13 to E-D-44, conditions: a) SOP 32 a/b for X SMe/ Sores or SOP 33 for X OTCA 70 for E-D-23, ax/J mixture); b) 1. SOP 9; 2. SOP 00 Compound E-D-27: E-D-27 was formed according to SOP 33 with ether as solvent at -20 0 C and TBDMSOTf as promotor in 70 yield j3-mixture).

S 5 Selected 1 H-NMR (400 MHz, CDCI 3 7.58 (in, 2H, Aryl), 7.54 (in, 2H, Aryl), 7.36 7.00 (in, 23 H, Aryl), 6.73 (in, 2H, Aryl), 5.37 1 H, J 1 2 3.9 Hz, 5.12 (dd, 1 H, J 2 3 8.8 Hz, 4.63 1 H, Jgem 11.2 Hz, OCH 2 4.58 1IH, Jgem 11.2 Hz, OCH 2 4.48 1 H, J 12 7.3 Hz, H-1 3.66 (s, c-i 3 H, OCHA) 3.55 3 H, OCH 3 3.34 (mn, 1 3.22 (dd, 1 H, J 3.4 Hz, J= 00 c) 10 10.7 Hz), 1.81 3 H, OAc), 0.98 9H, C(CH 3 3 Example 20 :Synthesis of alternative E-D-d isaccha rides E-D-45 to OBz OR 1 BnO 0& HO-Q BnO X.'mnb -X HBnO OTBOPS R BnO E-1, E-5, E-1-iv-a, D-8a: R 1 Mp E-1,.iv-b, E-5-ii-a, E-5-ii-b D-8b: R 1 =Mpm I D-8c: R 1 =All OBz Ia BnO 0Y 10R2

OR

1 BnO 0 OTBDPS BnO R 1= Mp, R 2= N 3 E-D46 R1 =MR2 NDP E-D-46: Ri Mp, R 2= N3DP E-D-47: R I=Mpm, R 2 NHD E-D-48: R' All, R NDP E-D-50: R1 All, R 2= NHDTPM 00 Example 20 Synthesis of alternative E-D-disaccharides E-D-45 to conditions: a) SOP 32a/b for X SMe/SCres or SOP 33 for X OTCA, diethyl ether, TBDMS-OTf, -20 deg. C (75 for E-D-45 as ax/j-mixture).

00 00 Example 21: Synthesis of trisaccharides E-D-C-1 to E-D-C-16 on R OMe OR'

HO~

0 -4..OTBDPS E-D-9 E-D-1O B nO OTCA N 3 C-1, /C-7 I C-13 IC-14

OR

3 BnO- Bno R 0 OMe ORI

R

2

OTBDPS

BnO N 3 E-D-C-1: R 1 Mpm, R 2 Mpm,R 3 =Mpm, R"=NHDTPM E-D-C-9:R I Mpm, R 2 =Mpm, R 3 Mpm, R 4

N

3 E-D-C-2: R 1 Mp, R 2 Mpm, R 3 Mpm, R 4 =NHDTPM E-D-C-1O: R1Mp, R 2 Mpm, R 3 Mpm, R 4

N

3 E-D-C-3: R I Mpm,R 2 =Mpm,R 3 =Bz, R 4 NHOTPM E-D-C-11: R 1 =Mpm, R 2 =Mpm, R 3 =Bz R 4

=N

3 E-D-C-4: R1 Mp, R 2=Mpm, R 3= Bz, R 4=NHDTPM E-D-C-12:R1 Mp, R 2=Mpm, R 3 =Bz, R 4

=N

3

R

1 =Mpm, R 2 =Bz, R 3 Mpm, R 4 NHDTPM E-D-C-13: R 1 =Mpm, R 2 =Bz, R 3 Mpm, R 4 N 3 p 2 =zR3 PR4 HTMEDC 14:R p 2 =zR3 ,R4 =N E-D-C-7: R1 Mp, R 2 5z, R 3= Mpm, R 4= NHDTPM E-D-C-14: R I= Mp, R 2 Bz, R 3= Mpm, R 4= N, E-D-C-8: RI Mp, R 2 =Bz, R 3 =Bz, R 4 NHDTPM E-D-C-15: R 1 Mp, R 2 Bz, R 3 Bz, R 4

N

3 Example 21: Synthesis of trisaccharide E-D-C-1 to E-D-C-16, conditions: a) SOP 33, (70 for E-D-C 15 as an a/p3 mixture).

Compound was formed according to SOP 33 with dichioromethane as solvent at 0 to 200C and TBDMSOTf as promotor in 70 yield (ax/j-mixture).

1 H-NMR (400 MHz, 00013):, 5= 7.93 (in, 2H, Aryl), 7.87 (in, 2H, Aryl), 7.66 (in, 2H, Aryl), 7.61 (in, 2H, Aryl), 7.46 (mn, 2H, Aryl), 7.38-6.99 (in, 32 H, Aryl), 6.79 (mn, 2H, Aryl), 5.27 1 H, J 1 2 3.8 Hz, H-1"cc), 4.99 (dd, 1 H, J 3 4 9.5 Hz, 4.80-4.69 (in, 6 H, 00H 2 4.52 (in, 3 H, 00H 2 4.40 1IH, J1, 2 8.0 Hz, H-1 4.38-4.32 (in, 2 H, not assigned), 4.29 1 H, J 1 2 Hz, H-1'03), 4.15 (in, 1 H, Jge m 12.0 Hz, OCH 2 4.02 (dd, 1 H, J 4 5 9.6 Hz, 3.80 (2 dd, 2 H, 3.71, 3 H, 00H 3 3.67 (in, 1 H, not assigned), 3.61-3.53 (in, 2 H, H5', 3.46 (dd, 1 H, Jgem =11.2 Hz, J 5 ,6a 2.4 Hz, H-6a), 3.41 (dd, 1 H, J 2 3 J34= 9.0 Hz, 3.27 3 H, 00HA) 3.21 (d d, 1 H, J 2 3 10.0 Hz, 3.14 (dd, 1 H, 3.00 (dd, 1 H, J 5 6 b Hz, H-6b), 2.75 (mn, 1 H, H-5) 1.05 9H,C(0H 3 3 Example 22: Synthesis of trisaccharides E-D-C.17 to E-D-C-32

OR'

BnO 0~O BnO4-t E-D-37 to E-D-4 Bn~.

B.O

9

VTCA

OR'

HO

2 OOTBDPS

N

3 CA-I C-7 IC,9 I C-1 R. OMe

OR'

BzO N 3 E-D-C-17: R1 Mpm, R 2= Mpm, R 3 Mpm, R 4

=NHDTPM

E-D-C..18: R 1 =Mp, R 2= Mpm, R 3= Mpm, R NHDTPM E-D-C-19: R1 Mpm, R 2 Mpm,R 3 Bz, R 4

=NHDTPM

1 MP, R 2 Mpm,R 3 Bz, R 4 NHDTPM E-D-C-21: R 1 Mpm, R 2 =Bz, R 3 Mpm,R 4

=NHDTPM

E-D-C-22: R1 Mp, R 2 =Bz, R 3 Mpm,R 4 NHDTPM E-D-C-23: R 1 =Mpm, R 2 Bz, R 3 Bz, R 4

NHDTPM

E-D-C-24: R I= Mp, R 2 =Bz, R 3 =Bz, R 4

=NHDTPM

E-D-C-25: R 1 Mpm, R 2 =Mpm, R 3 Mpm, R 4

=N

3 E-D-C-26: R 1 Mp, R 2 Mpm, R 3 Mpm, R' 4 N 3 E-D-C-27: R 1 Mpm, R 2 Mpm, R 3 Bz, R 4

N

3 E-D-C-28: R 1 Mp, R 2 Mpm, R 3 Bz, R 4 N 3 E-D-C-29: R 1 =Mpm, R 2 8Bz, R 3 Mpm, R 4 N 3 E-D-C-30: R 1 MP, R 2 =Bz, R 3 Mpm, R 4 N3 E-D-C-31: R 1 Mpm, R 2 =Bz, R 3 Bz, R 4

=N

3 E-D-C-32: R 1 Mp, R 2 9Bz, R 3 Bz, R 4 N 3 Example 22: Synthesis of trisaccharide E-D-C-17 to E-D-C-32, conditions: a) SOP 33.

Example 23: Formation of trisaccharidic Trichioroacetimidates E-D-C-33 to E- D-C-48

OR

3 Bno o4 0:OMe

OR'

BnO N, E-D-C-1 to E-D-C-16 E-D-C-33: R 1 =Mpm, R 2 Mpm, R 3 Mpm, NHDTPM E-D-C-34: R 1 =Mp, R 2 Mpm, R 3 Mpm, R 4

NHDTPM

R

1 Mpm, R 2 Mpm, R 3 =Bz, R 4

=NHDTPM

E-O-C-36: R 1 MP, R 2 Mpm, R 3 Bz, R 4

NHDTPM

E-DC-7:R1= PM R2 =BR3 4pR =ND E-D-C-37: R 1 Mp, R z, R Mpm, R NHDTPM E-D-C-38: R 1 Mp, R 2 =Bz, R 3 Mpm, R 4

=NHDTPM

R1 Mo. R 2=Bz. R 3 Bz. R 4

=NHDTPM

E-D-C-41 :R 1 =Mpm, R =Mpm, R 3 =Mpm,R 4

=N

3 E.D.C42: R 1 =Mp,R 2 =Mpm, R 3 =Mpm, R 4

=N

3 E-D-C-43: R 1 =Mpm, R 2 =Mpm, R 3 =Bz,R 4

=N

3 E-D.C-44: R 1 =Mp, R 2=Mpm,R 3=Bz, R 4=N 3 E-D-C-45: R 1 Mpm, R 2 =Bz, R 3 Mpm, R 4

=N

3 E-D-C-46: R 1 =Mp, R 2 Bz, R 3 =Mpm, R 4

=N

3 E-O-C-47: R 1 Mpm, R 2 Bz, R 3 Bz, R 4

N

3 E-D-C-48: R 1 =Mp, R 2 =Bz, R 3 =Bz, R 4

=N.

Example 23: Formation of trisaccharidic Trichioroacetimidates E-D-C-33 to E- D-C-48, conditions: a) 1. SOP 9; 2. SOP 25, (82% over 2 steps for E-D-C-47 2008200567 07 Feb 2008 Example 24: Syntheses of trisaccharides E-D-C-9 to E-D-C-12 and E-D-C-49 to o OMe ievo-

A

B.D.3 BzO

OTCA

OR'

HO RQL R

N

3 C-3 to C-9 C-6a:R =SCres, R 1 =Mpm, R 2 Mpm C-7:R =OTBDPS, R I Mpm, R 2 Mpm C-8c: R =OTBDPS, RI Mp, R 2=AI C- 0:R =SCres,R I Mp, R 2 =Mpm C-11 :R =SCres, R 1 Mpm, R 2

=AJI

C-12: R =OTBDPS, R I=Mpm, R 2=AJI C-13:R =SCres, R1 Mp, R 2 All C-14:R =OTBDPS, R I=Mp, R 2= Mpm D-C-5: R OTBOPS, R 1 Mpm, R 2 Mpm D-C-6: R OTBDPS, R I Mpm, R 2 =AJl D-C7:R OBDP, I M,1 2= M D-C-8: R OTBOPS, R1 Mp, R 2= M D-C-1 R SCres, RI Mpm, R 2

=MPM

D-C-2:R =SCres, Ri Mpm, R 2= AJl D-C-3: R =SCres, R Mp, R 2=Mpm D-C-4:R =SCres, R Mp, R 2=All 0 OMe OR 1 BzON OR 3 b

R

13 OR 3 BnO B0 BzO RO2,

N

3 E-D-C-49: R =SCres, RI Mpm, R 2=Mpm, R 3=Bz =S~es I pR2 =lR3 =B R SCres, RI Mp, R A11, R3 Bz E--C52 =S~es 1 =MR2 JR3 :R =SCres, R Mp,R 2=Mpm, R 3=M E-D-C-52: R SCres, R 1 Mp, R 2 A, R 3 BzM E-D-C-53: R SCres, R 1 Mp, R 2 =Mpm, R 3 =Mpm E-D-C-56: R SCres. R I M o. R 2 All. R 3 =MoM E-D-C-1lI R =OTBDPS, RI Mpm, R 2 Mpm, R 3 Bz E---7:R=OTDS 1 p ,R2 lR3 E-D-C-57: R OTBDPS, R1 Mp, R pl, R 3= Bz E-D-C-12: R OTBDPS, R 1 Mp, R 2 =MAl, R 3 Bz R =OTBDPS, R1 Mpm, R 2=AI, R 3=zP E-D-C-59: R OTBDPS, R 1 =Mpm, R 2 Mp, R 3 MpM E-D-C-19: R OTBPS, R 1 Mp, R 2 Al, R 3 Mpm E-D-C-1O: R OTBPS, R1 Mp, R2 All, R MpM Example 24: Syntheses of trisaccharides E-D-C-9 to E-D-C-1 2 and E-D-C-49 to E-D-C-60, conditions: a) 1. SOP 33; 2. SOP 24; b) SOP 33.

(for D-C-5: 70 2 steps); b) SOP 33, 78 for E-D-C-9 as an Wc/ mixture).

99 00 Compound was formed according to SOP 33 with ether as solvent at -200C and TMSOTf as promotor, followed by SOP 24 in 70 yield (over 2 steps as c/13mixture).

S 5 Selected 1 H-NMR (400 MHz in CDCI 3 13 7.88 (in, 2H, Ar), 7.67- 7.58 (in, Ar), 7.42 (mn, 2H, Ar), 7.37-7.12 (mn, 16H, Aryl), 6.84 (in, 3H, Ar), 5.14 (dd, 1 H, J 1 2 8.2 Hz, J 2 3 9.5Hz, 4.90 1 H, Jgem 10.7 Hz, 00H 2 4.73 1 H, Jgem 11 .5 Hz, OCH 2 4.65 1 H, J 1 2 8.2 Hz, H-i 4.63 ci 4.58 (in, 2H, OCHA) 4.51 1 H, Jgem =12.0 Hz, OCH 2 4.20 1IH, J 12 00 7.9 Hz, H-i13) 4.05 1 H, Jgem 11 .9 Hz, 00H 2 4.02 3.95 (in, 2H, not c-iassigned), 3.81 3H, OGH 3 3.80 3H, OCH 3 3.71 1 H, J 4 5 9.9 Hz, 3.67 3H, OCH 3 3.47 3.40 (in, 3H, not assigned), 3.21 (dd, 1 H, J 9.0 Hz, J 9.8 Hz, not assigned), 3.00 (dd, 1 H, J 5 6 b 1.4 Hz, Jgem 10.5 Hz, H-6b), 2.63 (in, 1 H, 2.35 (bs, 1 H, 4-OH), 1.07 9H, C(CH 3 3 Comp~ound E-D-C-9: E-D..C-9 was formed according to SOP 33 with ether as solvent at -20'C and TBDMSOTf as promotor in 78 yield (cc/j3-iixture).

Selected 1 H-NMR (400 MHz, CDCI 3 7.77 (in, 2H, Aryl), 7.59, 7.54 (2m, 2 x 2H, Aryl), 7.35 7.00 (in, 30 H, Aryl), 6.88 (in, 2H, Aryl), 6.82 (in, 2H, Aryl), 6.73 (in, 2H, Aryl), 5.41 1 H, J 1 2 3.5 Hz, H-1"cc), 5.19 (dd, 1 H, J 2 3 ;zJ, 9.6 Hz, 4.85 4.78 (in, 4 H, OCHA) 4.67 (in, 2H, OCHA) 4.65 1 H, JI2= 8.5 Hz, H-1 0, not assigned), 4.38 1 H, Jgem 11. 1 Hz, OCH 2 4.29 1 H, Jgem 11.7 Hz, OCH 2 4.17 (dd, 1 H, not assigned), 4.11 1 H, J 1 2 7.9 Hz, H-113 not assigned), 4.03 1 H, Jgem 12.0 Hz, OCH 2 3.90 3.76 (in, 3H, not assigned), 3.730, 3.727 (2s, 2 x 3H, OCHA) 3.65 3H, OCH 3 3.54 3H, OCHA) 2.89 (dd, 1 H, Jgem 10.5 Hz, J 5 6 b 2.0 Hz, H-6b), 2.52 (in, 1 H, 1 .02 9 H, C(CH 3 3 Example 25: Synthesis of trisaccharides E-D-C-41, E-D-C-42 and E-D-C-61 to E-D-C-66

)MPM

E-D-C-53: E-D-C-54: E-D-C-56: R1= Mpm, R 2=Mpm Mpm, R 2 =All Mp, R 2=Mpm MP, R 2 =All 13 E-D-C-61: R' Mpm, R E-D-C-62: R1= Mpm, R E-D-C-63: Ri= Mp, R 2 E-D-C-64: Ri= Mp, R 2 E-D-C-41: RI Mpm, R E-D-C-42: R' Mp, R 2 R1 Mpm, R E-D-C-66: RI= Mp, R 2 BnO 0~p

N

3 1 0 Me OR 1 2 All00 2Mp BO R N =:All OMpm b

N

:Mpm 0n 2=All BnO R All N 3 Example 25: Synthesis of trisaccharides E-D-C-41, E-D-C-42 and E-D-C-61 to E-D-C-66, conditions: a) 1. SOP 39; 2. SOP 38; 3. SOP 16; b) 1. SOP 14; 2.

SOP Example 26: An alternative route to the trisaccharides E-D-C-61 and E-D-C- D-C-9: R 1 =Mpm D-C-1: R1 Mp D-C-11 :R 1 Mpm D-C-12: R 1 Mp BnO' Br O 0e.OMe Bn Mpm O ~imm&.SCres BzO

N

3 b OMpm BnO BnO 0 OMe E-D-C-61

:R

1 Mpm N

R

E-D-C63: R 1 Mp BOm4 BnO MPMO SC res

N

3 Example 26: An alternative route to the trisacoharides E-D-C-61 and E-D-C- 63, conditions: a) 1. SOP 39; 2. SOP 38; 3. SOP 16; 4. Pd(Ph 3

P)

4 p- ToISO 2 Na, THE, MeOH; b) SOP 33.

Example 27: Svntheses of blocks E-D-C-67 to 00 o OMe

M

LevO +HOO1 0~~S~e BnO OTCA re AllocO NH D-9I aI 0 0 OMe OMp BnO SC res AllocO D-C-1 3 NH 00b o OMe OMp HO-\ 0 BnO 0SCres Alloco D-C-14 NH

OR

BnO- 0 BnO+ IE-I E-3 N 3 OC

C

OR OC BnO 0~O BflO4 m 0 OMe OMp BnO 0SCres Z ;NH E-D-C-67: R Mp AllocO

N

E-D-C-68: R Bz 0 ORd BnO- 0 BnO o OMe OMp 030 BnO SC res u NH E-D-C-69: R Mp BnO

N

R Bz 0 Example 27 Syntheses of trisaccharides E-D-C-67 to E-D-C-70, conditions: a) SOP 33; b) SOP 24; c) SOP 33; d) 1. SOP 36; 2. SOP 37.

Example 28: Synthesis of trisaccharides E-D-C-70 and E-D-C-71 C-1 R SCres E-D-C-71 :R OTBDPS Example 28: Synthesis of trisaccharides E-D-C-70 and E-D-C-71, conditions: a) SOP 33, (55% for E-D-C-71, cx/p3 mixture).

Compound E-D-C-71: E-D-C-71 was formed according to SOP 33 with dichioromethane as solvent at 40 0 C and TBDMSOTf as promotor in 55 yield (as acf3-mixture).

Selected 1 H-NMR (400 MHz, CDCI 3 9= 7.91 (in, 2H, Aryl), 7.61 (in, 2H, Aryl), 7.55 (in, 2H, Aryl), 7.50 7.02 (in, 29 H, Aryl), 6.65 (in, 4H, Mp), 5.38 (d 1 H, J1, 2 3.9 Hz, 5.22 (bs, 1 H, NH), 4.67 1 H, J 1 2 7.4 Hz, H- 103, not assigned), 4.50 1 H, J 1 2 7.8 Hz, H-1 0, not assigned), 3.92 1 H, 9.8 Hz, 3.698 3 H, OCH 3 3.693 3 H, OCH 3 1.03 9 H,

C(CH

3 3 Mfound 1408.52 (M+H 2 Mcalc 1390.54 Examrle 29: Syntheses of trisaccharides E-D-C-61, E-D-C-72 and E-D-C-73 o Me LevO 0~~ BnO m. OTCA BnO OMpm

HO~

SCres

N

3 C -6-a 0~ MPMO SCres I-C-1 5 N 3 OMpm MOPMO m~ SCres

C

E-1 to E-3 E-D-C-61: R Mpm Bi E-D-C-72: R Mp E-D-C-73: R Bz Example 29: Syntheses of trisaccharides E-D-C-72 to E-D-C-73 and E-D-C- 61, conditions: a) SOP 33; b) SOP 24; c) SOP 33.

Example 30: Syntheses of trisaccharides C-B-A-I to C-B-A-4 00 00 O13z C-17

N

3

OTCA

a 0R ~SCres HO B-1 O13z 'SCres LevO'

AMH

OMe C-B-A-I: R- N C-B-A-2: R= DPMNH OMe O13z C-B-A-3- R-N C-B-A-4: R=PMNH Example 30: Syntheses of trisaccharides C-B-A-I to C-B-A-4, conditions: a) SOP 33; b) SOP 32a; c) 1. SOP 27; 2. SOP 20; 3. SOP 16; 4. SOP 24.

00 Example 31: Synthesis of trisaccharides C-B-A-5 to C-B-A-8 OMpm O13z 0 Me0 RO -0N+

R

1 Ne TCA+ nW IDC-18:R =Bz, R =Ac 813 BA-9 C-19:R =Mpm,R =Ac O =Bz, RI Lev 00 C-21:R =Mpn, R Lev O13z OMpm MeO

R

1 0 RO 0 N3 e

N

3 03 0 =Bz, R 1 Ac C-B-A-6:R =Mpn, R1 Ac C-B-A-7:R =Bz, R1 =Lev C-B-A-B: R Morn, R 1 Lev Example 31: Synthesis of trisaccharides C-B-A-5 and C-B-A-8, conditions: a) SOP 33, (50% for C-B-A-5, a13 mixture).

Compound C-B-A-5 was formed according to SOP 33 with ether as solvent at 200C and TBDMSOTf as promotor in 50 yield (as Wf3p-mixture).

Mfound 1269.65 (M+H+H 2 Mcalc 1250.43 Example 32: Syntheses of D-C-B trisaccharides D-C-B-1 to D-C-B-3.

LevO 0~O BnO OC AllocO D-9 C-B-2, III Bz C-B-3, R I Mp C-B3-4, RI Mpm O13z D.C-B3-I, D-C-B-2, D-C-B-3, RI= Bz RI Mp RI Mpm Example 32: Syntheses of D-C-B-trisaccharides D-C-B-1 to D-C-B-3, conditions: a) 1. SOP 33; 2. SOP 36; 3. SOP 37.

Example 33: Syntheses of D-C-B trisaccharides D-C-B-4 to D-C-B-7.

oOMe BnO OC AllocO D-9 O13z C-13-5: R 1 Mpm, R 2 Mpm, C-13-6: Ri= Mpm, R 2 All, C-13-7: R1 Mp, R 2= Mpm, C-B-8: R' Mp,R 2=All, MeO0B 0 7jLO1' SCres D-C-B3-4: R1= Mpm, R 2 Mpm, Ri Mpm, R 2= All, D-C-B-6: R1= Mp, R 2 Mpm, D-C-B3-7: R1= Mp, R 2 All, 00 Example 33: Syntheses of D-C-B-trisaccha rides D-C-B-4 to D-C-B-7, conditions: a) 1. SOP 33; 2. SOP 36; 3. SOP 37.

(NO

00 109 Exarle 34: Syntheses of tetrasaccha rides D-C-B-A-1 to D-C-B-A-2 D-6a 34: Syntheses of tetra saccha rides D-C-B-A-1 and D-C-B-A-2, conditions: a) SOP 32a; b)1. SOP 36; 2. SOP 37; 3. SOP 24.

Example 35: Alternative syntheses of tetrasaccharides D-C-B-A-2 .OMp D-C-B-A-2 Example 35: Alternative synthesis of tetrasaccharide D-C-B-A-2, conditions: a) 1. SOP 34; 2. SOP 24.

Examr~Ie 36: Syntheses of tetrasaccha rides D-C-B-A-3 to D-C-B-A-8 R' Mpm, R 2=Mpm D-C-17: R1 Mpm, R 2=MP D-C-18: RI Mpm,R 2 =Bz a O OMe OR 2 HO00 BnO 1 BnO N 3 D-C-B-A-3: R= DTPMNH, RI Mpm, R 2 Mpm, D-C-B-A-4: R= DTPMNH, R 1 Mpm, R 2 Bz R= DTPMNH, R 1 Mpm, R 2 =Mp D-CB-A6: N, 1 pR2 =M D-C-B-A-6: R= N 3 RI Mpm, R 2= Mpm D-C-B-A-8 R= N RI M pm, R 2

M

-f OMe OBz B-A-9: R N B-A-la: RLTrPMNH Example 36: Syntheses of tetrasaccharides D-C-B-A-3 to D-C-B-A-8, conditions: a) 1 SOP 32b; 2. SOP 24.

Example 37: Syntheses of tetrasaccharides E-D-C-B3-1 to E-D-C-B-4

OR

BnO 0I-O BnO~m o OMe

N

3 BnO OC E-D-29: R =Bz OAIIoc E-D-31: R =Mpm r-SCres Bz BnO,-

C

Bn E-D-C-B-1 R Bz E-D-C-B-2: R Mpm E-D-C-B-3: R Bz E-D-C-B-4: R Mpm OBz Example 37: Syntheses of tetrasaccharides E-D-C-B-1 to E-D-C-B-4, conditions: a) SOP 33; b) 1. SOP 36; 2. SOP 37.

Example 38: Syntheses of blocks E-D-C-B-5 to E-D-C-B-8 E-D-29: R Bz E-D-51 :R Mp R Bz E-D-C-B-6: R Mp BnO E-D-C-B-7: R Bz E-D-C-B-8: R Mp Example 38: Syntheses of tetra saccha rides E-D-C-B-5 to E-D-C-B-8, conditions: a) SOP 33; b) 1. SOP 36; 2. SOP 37.

Example 39 Syntheses of E-D-C-B-A pentasaccharides P-1 and P-2 E-D-C-69: R Mp R Bz OBz 0 B"-A-9N 3 0 W~e HO OBz P-1 R Mp P-2: R= Bz Example 39: Syntheses of E-D-C-B-A pentasaccha rides P-I and P-2, conditions: a) SOP 34.

Examole 40: Synthesis of E-D-C-B-A oentasaccharides P-3 to P-26 00 R2 E-D-C-41 to E-D-C-48 R E-D-C-65 to E-D-C-66 B-A-9 BnO E-D-74 to E-D-C-87 B Bn 0Me MO OqBn COnO nO R+

N

3 O 0Me~O' 8 8 n nO 03 Me-0M BnnO

R

(--KiBn P-:R IIR MeR Mp Me 6 z P-3R=AI,3 Mpf 2 B -i RMmR p R 2 z P1:~, 1 mR Bz 1n 2 3 2M1 e P-3: R =All, RI Mp,R2 =Bz P-11 :R =Mpm,RI Mp,R2 Bz P-19: R=Bz,RI Mp,R2 Bz P4:R =All M, R2 =z P.12: R =Mpm, R I Mp,R MpmB P20: :R =Bz, R I =Mp,R pm 005 R lRI=Mm 2 1 p -3 Mm '=Mm 2 1 z '=Mm 2= M 00 P-6: R =All, R =Mp R2 MpM P-14:R =Mpm, R Mp,R 2=Mpm P-22: R =Bz, R' MP,R 2=Mpm 0 R =All,R 1 =-Mp R 2 Mp P-1 5:R =Mpm, R 1 MpR 2 MP P-23: R =Bz, R' =Mpm, R 2

MP

P-8: R=All, R I=Mpm,R2 Mp P-16: R =Mpm, R I =Mpm, R 2 =MP P-24: R =Bz, R Mp,R2 Mp P-9: R =All, R =Mpm,R2 TBDPS P-117: R =Mpm,RI Mpm, R 2=TBDPS P-25: R =Bz, R Mpm, R 2=TBDPS 1 =All, RI Mp, R2 TBOPS P-1 8:R =Mpm, RI=Mp, R 2 =TBDPS P-26: R R' Mp, R 2

=TBDPS

Example 40: Synthesis of E-D-C-B-A pentasaccharide P-3 to P-26, conditions: a) SOP 33 (75 for P-19 as an c/f3 mixture).

Compound P-19: P-19 was formed according to SOP 33 with dichioromethane as solvent at 2000 and TMVSOTf as promotor; Mfound 2068.76 (M+H+H 2 0) 4 Mcalc 2049.74 ExamDle 41: Alternative syntheses of E-D-C-B-A Dentasaccha rides P-1I1.

P-12. P-19. P-20 and P-27 OBz A-I N 3 1 OMe OBn N 3 E-D-C-B-3: R Mpm, R 1 Mpm E-D-C-B-9: R Mpm, R' Bz, E-D-C-B-1O: R Mpm, R 1 Mp E-D-C-B-I1 :R Bz, R I MP E-D-C-B-12: R Bz, R I Mpm OBn N P-11: R =Mpm, R Mpm P.12:R =Mpm,R =Mp P-19:R =Bz, R Mpm =Bz, R =Mp P-27: R =Mpm R Bz Example 41: Alternative syntheses of E-D-C-B-A pentasaccharides P-11, P- 12, P-19, P-20 and P-27, conditions: a) SOP 32a.

Example 42: Alternative syntheses of some E-D-C-B-A pentasaccharides OR 2 BnO R E-I to E-3, N 3 E.1 -iv-b, E-I-v-b, E-1 -vi-b P-I1 P-12: P-13: P-14: R' Mpm,R =Bz R1 =MR2 =B R I=Mp,R 2=zP R1 =MR2= M R I=Mp, R 2=Mpm P-16: R Mpm, R=Mp P-27: R' Bz, R 2 Bz P-28: R BzR=Mp P-29: R' Bz,R 2

=MPM

Example 42: Alternative syntheses of some E-D-C-B-A pentasacoharides, conditions: a) SOP 32a (for R SCres) or SOP 33 (for R OTCA).

Example 43:Synthesis of pentasaccharide P-1 3 and OMpm

N

3 0 OPe E-D-C-49 ~Op BnO MPMO.4.' O. SCres

N

3 OBz MeOO~ 0 BnO.A 0 B.A9,RI~ F T B-A-laO0 HO OBz P-13: R N 3 R DTPMNH Example 43: Formation of pentasaccha rides P-13 and P-30, conditions: a) SOP 32a.

Example 44: Formation Of Dentasaccharide P-19 and P-31 OBz Bn0O i I R 0 O 0 E-D-7 E-D-10O F n

C

BnOnTC P.19: R=N P-31: R=N DTPM Example 44: Formation of pentasaccharide P-19 and P-31 conditions: a) SOP 33 Example preparation of P19: 00 00 A mixture of O-(2-azido-6-O-benzoyl-3 ,4-di-O-benzyl-2-deoxy-ac-D-glucopyranosyl)-( 1 -+4)-(methyl 2 ,3-d i-O-benzyl-f3-D-glucopyranosyluronate)-( 1 azido-3-O-benzoyl-2-deoxy-6-O-p-methoxybenzyl-a-Dgluco pyra nosyltrichloroaceti mid ate (30.0 mg, 21.2 tmol) and methyl (methyl 2-O-benzoyl-3-O-benzyl-ct-L-id upyranosyluronate)-( I -+4)-2-azido-3-O-benzyl- 6-O-benzoyl-2-deoXy-ct-D-glucopyranoside (15.4 mg 19.3 pImol) and 100 mg of molecular sieves 4A in 1.5 ml dry dichioromethane was treated with TBDMSOTf (0.97 [id, 4.24 4imol) at -20 0 C for 20 hours. The reaction was quenched, filtered and concentrated. Further purification of the title compound was achieved by silica gel chromatography Yield: 15.83 mg Rf= 0.30 (toluene/ethyl acetate 9/1).

Example 45: Partial deprotection of Pentasaccharide P-19 Example 45: Partial deprotection of pentasaccharide P-19, conditions: a) SOP 28, 84%; b) SOP 39, 86%.

Compound P-33: Mfo~fld 1503.5 (M-N 2 Mcalc 1529.51 To ease the structural proof, a small part of P-33 was transformed into the bis methyl uronurate derivative and characterized via NMR-spectroscopy.

Characteristic 1 H-NMR-spectral regions are shown in figure 1.

Mfound 1514.62 Mcai 1513.58 Example 46: Partial deprotection of pentasaccharide P-30, containing a DTPM-arouo as amino orotection P-33 Example 46: Partial deprotection of pentasaccharide P-30, containing a DTPM-group as amino protection, conditions: a) 1. SOP 28; 2. SOP 39; b) 1.

SOP 11 with MeNH 2 as primary amine and MeOH as solvent; 2. SOP 12.

Example 47: Partial deprotection of pentasaccharide P-1, containing a cyclic carbamate as amino protection 00 OMp C0 OBz BnO OMp MeO OMe N K O N, cI BnO OMe OBn NHOB OH lbz BnO 0

OH

OOHO

IND Me ~eO OH O 0 OHB I~

OH

Cn OH BnO~o z N]o OaNaO 0 BnO BnO HO3 OMe OBn NH 0 OH Example 47: Partial deprotection of pentasaccharide containing a cyclic carbamate as amino protection, conditions: a) SOP 27; b) SOP 39; c) SOP 12.

Example 48: Derotection protocol for pentasaccharides P-37 of claim 4 00 ORss ORSi C- O O R ORE 2 RHO RH2Z o ORE1

OR

3

RRHO

RBI

RH= RHe .RH 2 RPl RP=0 OR 0= R 5

R

53 R R 55 Mpm, Mp, Bz; RA Re =RB 1 NHDde, NHDTPM, N 3 or RS4 and RB cyclic Carbarmate; a REI RE2 Me, All, Bn; X a-OMe 00

OH

C)BnO 0~

OH

C Bn ONa OH ONa 0 03 BnO N 3 00O M e P-33 OBnN 3

O

0 Example 48: Deprotection protocol for pentasaccharides P-37 of claim 4, conditions: a) 1. SOP 27 and 28; 2. SOP 39; 3. SOP 11 with MeNH 2 as primary amine and MeOH as solvent; 4. SOP 12.

Example 49: Transformation of pentasaccharide P-33 into the 0-and Nsulfated pentasaccharide 00 OH 0

OH

BnO

N

3 1 0OH NaO BnO Bn OMe

OSO

3 Na BnO H io OH ,soN BnO7 bS Ia 0OSO 3 Na Bn 0 ONa0

N

3 OSO 3 Na NaO OBn 0 Bn N3 BnO P-38 N3 OMe I OS03Na BnO NOS HOi NN OS03Na H 0 0 OS03Na 00 H 2 N 0 ONa O 0~1 OS03Na NaOH0 O I"r OH

H

H O00 P-'39 OMe

OSO

3 Na HO NaO 3 SO~ H2 OS 3 Na 0N 0 OSO 3 Na HO I NaOSO 2 NH 0 O-a OSO 3 Na NaO OH HO 0 -0 ~NaOSO 2

NHI

OMe HO NaO 3 SO NaOSO 2 NH OSO 3 Na Example 49: Transformation of pentasaccharide P-33 into the 0-and Nsulfated pentasaccharide P-40, conditions: a) SO 3 xNMe 3 DMF, 5000; b) H 2 (70 psi), Pd/C, H 2 0; c) SO 3 xPyridine, H 2 0, pH 9.5. The transformation of P- 33 into P-40 has been performed according to literature: Petitou et al., Carbohydr. Res. 1987, 167, 67-75.

The 'H-NMR (400 MHz, D 2 0) of P-40 is shown in figure 6.

Claims (8)

1. 3. A trisaccharide building block of claim 1, Wherein: X 2 is selected from a hydroxyl group; thioalkyl, thioaryl, chloro, fluoro, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta; 125 00 Rs3 is selected from the group consisting of 4-methoxyphenyl; Ssubstituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted ,0 alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting Sgroups; tert-Butyldiphenylsilyl; Rs 4 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted t alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting 0 groups; tert-Butyldiphenylsilyl, or RS 4 and RB may be combined to form a 00 10 cyclic carbamate; RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RS 4 and RB can combine together to form a cyclic carbamate; RP2 is selected from the group consisting of 4-methoxyphenyl; benzyl; substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups; carbonate protecting groups including allyloxycarbonyl; Rp1 is selected from the group consisting of 4-methoxyphenyl; benzyl; substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups; carbonate protecting groups including allyloxycarbonyl; REI is selected from the group consisting of methyl, C2-C5 alkyl; substituted alkyl; or, benzyl and substituted benzyl groups; RBI is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RH2 and RB1 can combine together to form a cyclic carbamate; 00 RH2 is selected from the group consisting of benzyl or substituted O 0 benzyl protecting group, or RH2 and RB1 independently can combine together Sto form a cyclic carbamate; RH is selected from the group consisting of benzyl or substituted benzyl protecting group; NO I Rss is selected from the group consisting of 4-methoxyphenyl; Ssubstituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted 00 10 alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting Sgroups; tert-Butyldiphenylsilyl;
2. 4. A trisaccharide building block of claim 1, Wherein: Rs 3 is selected from the group consisting of 4-methoxyphenyl; 4- methoxybenzyl, benzoyl, 4-chlorobenzoyl, allyl, allyloxycarbonyl; Rs4 is selected from the group consisting of 4-methoxyphenyl; 4- methoxybenzyl, benzoyl, 4-chlorobenzoyl, allyl, or RS 4 and Re may be combined to form a cyclic carbamate; RB is selected from the group consisting of an azido function, an amine, or Rs 4 and RB can combine together to form a cyclic carbamate; X2 is selected from a thiomethyl, thioethyl, thiophenyl, thiocresyl, trichloroacetimidoyl, tert-butyldiphenylsilyloxy and the stereochemistry may be alpha or beta; RE1 is selected from the group consisting of methyl, allyl, benzyl; is benzyl; Rp 2 is selected from the group consisting of benzyl; benzoate; or allyloxycarbonyl; RH RH2 is benzyl; is benzyl; Rss is selected from the group consisting of 4-methoxyphenyl; 4- methoxybenzyl, benzoyl, tert-butyldiphenylsilyl; RBI is selected from the group consisting of an azido function, or RH2 and RB1 can combine together to form a cyclic carbamate. A trisaccharide building block of General Formula XXIV, ORs2 General Formula XXIV (Block C-B-A) Wherein: X, is selected from the group consisting of hydroxy, alkenyloxy, alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, imidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid or other such group suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; RH and RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; 00 O RA is selected from the group consisting of an azido function, an Samine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Troc, N-phthalimido, T. NH-Ac, NH-Allyloxycarbonyl; or RH and RA can combine together to form a cyclic carbamate; Rsi is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted 0 alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting 00 10 groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting Sgroup allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; Rs 2 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; Rs 3 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl; Rs 4 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, or Rs 4 and RB may be combined to form a cyclic carbamate; RE2 is selected from the group consisting of methyl, C2-C5 alkyl; substituted alkyl,C 3 -C 5 alkenyl; or, benzyl and substituted benzyl groups; 129 00 RB is selected from the group consisting of an azido function, an 0 0 amine; an NH-Dde or NH-DTPM group, or Rs4 and RB can combine together :3 to form a cyclic carbamate; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4- acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or n other such suitable substituted benzyl type protecting group; y-aminobutyryl, S4-N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1 -ylidene)ethylamino]-butyryl, 4-N-[1- (1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, S4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; allyloxycarbonyl, allyl ether, carbonate type protecting groups;
3. 6. A trisaccharide building block of claim Wherein: Xi is selected from the group consisting of hydroxy, alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; RH and RH1 are independently selected from a benzyl or substituted benzyl protecting group, or RH1 and RA can combine together to form a cyclic carbamate; RA is selected from the group consisting of an azido function, an amine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Troc, N-phthalimido; or other such suitable protected amino functions or RH1 and RA can combine together to form a cyclic carbamate, 00 Rsi,RS2,Rs 3 and Rs 4 are independently selected from: 4-methoxyphenyl; 4- 0 O methoxybenzyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, benzoyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; or RS 4 and RB may be combined to form a cyclic carbamate; RE2 is selected from methyl, C2-05 alkyl; substituted alkyl; or, benzyl and substituted benzyl groups; 00 10 RB is selected from an azido function, an amine; an NH-Dde or NH-DTPM group; additionally RS 4 and Re can combine together to form a cyclic carbamate; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4- acetoxybenzoyl, 4-acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4- acetamidobenzyl or other such suitable substituted benzyl type protecting group; -aminobutyryl,y 4-N-[1 -(4,4-dimethyl-2,6-dioxocyclohex-1 ylidene)ethylamino]-butyryl, 4-N-[1-(1,3-dimethyl-2,4,6(1 trioxopyrimidin-5-ylidene)methylamino]-butyryl, 4-N-Alloc-butyryl, 4-N-Fmoc- butyryl, 4-N-Boc-butyryl type protecting group.
4. 7. A trisaccharide building block of claim Wherein: X1 is selected from the group consisting of hydroxy, methoxy, thiomethyl, thioethyl, thiocresyl, trichloroacetimidoyl, a tbutyldiphenylsilyloxy a lipoaminoacid suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; RH and RHI are independently selected from a benzyl or substituted benzyl protecting group, or RH1 and RA can combine together to form a cyclic carbamate; RA carbamate; is an azido, or RHI and RA can combine together to form a cyclic r- O 0 (N Rsi,Rs2,Rs 3 and Rs 4 are independently selected from: 4-methoxyphenyl; 4- methoxybenzyl; benzoyl, or Rs 4 and RB may be combined to form a cyclic 10 carbamate; RE2 is selected from methyl, allyl or, benzyl and substituted benzyl groups; RB is an azido function, or Rs 4 and RB can combine together to form a cyclic carbamate; is selected from a H atom; a levulinoyl.
5. 8. A tetrasaccharide building block of General Formula XXV, .ORsi RLO' RpO- ORp2 General Formula XXV (Block D-C-B-A) Wherein, Xi is selected from the group consisting of hydroxy, alkenyloxy, alkoxy, aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, imidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a 00 lipoaminoacid or other such group suitable for conjugation to delivery systems O Sor solid supports; and the stereochemistry may be alpha or beta, T. RH is selected from the group consisting of benzyl or substituted S 5 benzyl protecting group, allyl, allyloxycarbonyl; RH1 is selected from the group consisting of benzyl or substituted Sbenzyl protecting group, allyl, allyloxycarbonyl, or RH1 and RA can combine O together to form a cyclic carbamate; 00 RA is selected from the group consisting of an azido function, an amine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Troc, N-phthalimido, NH-Ac, NH-Allyloxycarbonyl; or RHI and RA can combine together to form a cyclic carbamate, Rsi is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, Rs2 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, Rs3 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, 133 00 Rs4 is selected from the group consisting of 4-methoxyphenyl; O substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted 3 alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, or Rs 4 and Re may be combined to form a cyclic carbamate, rO l REI is selected from the group consisting of methyl, C 2 -0 5 alkyl; 0 substituted alkyl, C 3 -C 5 alkenyl; or, benzyl and substituted benzyl groups; 00 RE2 is selected from the group consisting of methyl, C 2 -C 5 alkyl; substituted alkyl,C 3 -C 5 alkenyl; or, benzyl and substituted benzyl groups, RB is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or Rs 4 and RB can combine together to form a cyclic carbamate, Rp 1 is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; Rp 2 is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3- alkenyl; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4-acetoxybenzoyl, 4- acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4-acetamidobenzyl or other such suitable substituted benzyl type protecting group; y-aminobutyryl, 4-N-[1-(4,4-dimethyl-2,6-dioxocyclohex-l-ylidene)ethylamino]-butyryl, 4-N-[1- 00 (1,3-dimethyl-2,4,6(1 H,3H,5H)-trioxopyrimidin-5-ylidene)methylamino]-butyryl, O S4-N-Alloc-butyryl, 4-N-Fmoc-butyryl, 4-N-Boc-butyryl type protecting groups; allyloxycarbonyl, allyl ether, carbonate type protecting groups. c 9. A tetrasaccharide building block of claim 8, Wherein: \O In O X, is selected from the group consisting of hydroxy, alkoxy, 00 aryloxy, benzyloxy, substituted benzyloxy; thioalkyl, thioaryl, halogen, O trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; a lipoaminoacid suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; RH and RH1 are independently selected from a benzyl or substituted benzyl protecting group, or RHI and RA can combine together to form a cyclic carbamate; RA is selected from the group consisting of an azido function, an amine; an NH-Dde, NH-DTPM, NH-Fmoc, NH-Boc, NH-Troc, N-phthalimido; or RH1 and RA can combine together to form a cyclic carbamate; Rsi,Rs2,Rs3 and Rs 4 are independently selected from: 4-methoxyphenyl; 4- methoxybenzyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, benzoyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; or Rs 4 and RB may be combined to form a cyclic carbamate; REI and RE2 are independently selected from methyl, C 2 -C 5 alkyl; substituted alkyl; or, benzyl and substituted benzyl groups; 00 RB is selected from an azido function, an amine; an NH-Dde or NH-DTPM 0 O group; additionally Rs 4 and RB can combine together to form a cyclic 0 carbamate; RL is selected from a H atom; a levulinoyl, chloroacetyl, 4- acetoxybenzoyl, 4-acetamidobenzoyl, 4-azidobenzoyl, or other substituted benzoyl type protecting group; a benzyl group, a 4-acetoxybenzyl, 4- acetamidobenzyl or other such suitable substituted benzyl type protecting 0 group; -aminobutyryl,y 4-N-[1 -(4,4-dimethyl-2,6-dioxocyclohex-l- 00 ylidene)ethylamino]-butyryl, 4-N-[1-(1,3-dimethyl-2,4,6(1H,3H,5H)- 4-N-Alloc-butyryl, 4-N-Fmoc- butyryl, 4-N-Boc-butyryl type protecting group. A tetrasaccharide building block of claim 8, Wherein: Xi is selected from the group consisting of hydroxy, methoxy, thiomethyl, thioethyl, thiocresyl, trichloroacetimidoyl, a tbutyldiphenylsilyloxy a lipoaminoacid suitable for conjugation to delivery systems or solid supports; and the stereochemistry may be alpha or beta; RH and RH1 are independently selected from a benzyl or substituted benzyl protecting group, or RHi and RA can combine together to form a cyclic carbamate; RA is an azido function, or RH1 and RA can combine together to form a cyclic carbamate; Rs 1 ,RS2,RS3 and RS 4 are independently selected from: 4-methoxyphenyl; 4- methoxybenzyl; benzoyl, or RS 4 and Re may be combined to form a cyclic carbamate; REi and RE2 are independently selected from methyl, allyl or, benzyl and substituted benzyl groups; RB is an azido function, or RS 4 and RB can combine together to form a cyclic carbamate; is selected from a H atom; a levulinoyl.
6. 11. A tetrasaccharide building block of General Formula XXVI, ORs 2 E D C B General Formula XXVI (Block E-D-C-B) Wherein: RH is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl; RH2 is selected from the group consisting of benzyl or substituted benzyl protecting group, allyl, allyloxycarbonyl, or RH2 and RB1 independently can combine together to form a cyclic carbamate; Rs 2 is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; 00 O O Rs3 is selected from the group consisting of 4-methoxyphenyl; Ssubstituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted C alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl benzyloxymethyl; NO t RS4 is selected from the group consisting of 4-methoxyphenyl; Ssubstituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted 00 10 alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting Sgroups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl, or Rs 4 and RB may be combined to form a cyclic carbamate, Rss is selected from the group consisting of 4-methoxyphenyl; 4- methoxybenzyl, substituted benzyl groups; alkylacyl, benzoyl, arylacyl or alkylarylacyl, and substituted alkylacyl, 4-chlorobenzoyl, arylacyl or alkylarylacy protecting groups; allyloxycarbonyl, ethoxycarbonyl, tertbutoxycarbonyl, carbonate protecting groups; is selected from the group consisting of 4-methoxyphenyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, and substituted alkylacyl, arylacyl or alkylarylacy protecting groups; carbonate protecting groups; a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group allyl, methoxymethyl, methoxyethyl, benzyloxymethyl; or Rss and RH can be combined to form a cyclic acetal or ketal moiety; RE1 is selected from the group consisting of methyl, C 2 -0 5 alkyl; substituted alkyl, C3-C5 alkenyl; or, benzyl and substituted benzyl groups; RE2 is selected from the group consisting of methyl, C2-C5 alkyl; substituted alkyl,C 3 -Cs alkenyl; or, benzyl and substituted benzyl groups; 00 RB is selected from the group consisting of an azido function, an 0 O amine; an NH-Dde or NH-DTPM group, or RS 4 and RB can combine together CN Q3 to fonn a cyclic carbamate; RB1 is selected from the group consisting of an azido function, an amine; an NH-Dde or NH-DTPM group, or RH2 and RB1 can combine together to form a cyclic carbamate; SRp is selected from the group consisting of 4-methoxyphenyl; 00 10 benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or Ssubstituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups; RP2 is selected from the group consisting of 4-methoxyphenyl; benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl and alkylarylacyl protecting groups;, carbonate protecting groups, silyl protecting groups, carbamate protecting groups, C3- alkenyl; X 2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta
7. 12. A tetrasaccharide building block of claim 11, Wherein: RH and RH2 are independently selected from a benzyl or substituted benzyl protecting group, RH2 and RBI can combine together to form a cyclic carbamate; Rs 2 ,Rs 3 ,Rs 4 and Rss are independently selected from: 4-methoxyphenyl; 4- methoxybenzyl; substituted benzyl groups; alkylacyl, arylacyl or alkylarylacyl, 00 and substituted alkylacyl, benzoyl, arylacyl or alkylarylacy protecting groups; O Scarbonate protecting groups; or RS 4 and R 8 may be combined to form a Scyclic carbamate; 5 RE1 and RE2 are independently selected from methyl, C2-C5 alkyl; substituted alkyl; or, benzyl and substituted benzyl groups; NO RB and RBI are independently selected from an azido function, an amine; an 0 NH-Dde or NH-DTPM group; additionally RS 4 and Re or RH2 and RBI 00 10 independently can combine together to form a cyclic carbamate; Rpi and Rp 2 are independently selected from benzyl, substituted benzyl groups; alkylacyl, arylacyl and alkylarylacyl, or substituted alkylacyl, arylacyl, and alkylarylacyl protecting groups; and carbonate protecting groups; X2 is selected from a hydroxyl group; thioalkyl, thioaryl, halogen, trichloroacetimidoyl, phosphate and related phosphate ester type leaving groups, a tbutyldiphenylsilyloxy or other such substituted silyloxy protecting group; and the stereochemistry may be alpha or beta.
8. 13. A tetrasaccharide building block of claim 11, Wherein: RH and RH2 are independently selected from a benzyl or substituted benzyl protecting group, or RH2 and RB1 can combine together to form a cyclic carbamate; Rs 2 ,Rs 3 ,Rs 4 and Rss are independently selected from: 4-methoxyphenyl; 4- methoxybenzyl; benzoyl, or Rs 4 and RB may be combined to form a cyclic carbamate; REI and RE2 are independently selected from methyl, allyl or, benzyl and substituted benzyl groups; Re and Re 1 are independently an azido function, or (Rs 4 and Re) or (RH2 and n RBI) independently can combine together to form a cyclic carbamate; Rp is benzyl; Rp 2 is selected from the group consisting of benzyl; benzoyl, allyloxycarbonyl;. o 0 10 X 2 is selected from a hydroxyl group; thiomethyl, thiocresyl Strichloroacetimidoyl, or tbutyldiphenylsilyloxy; and the stereochemistry may be alpha or beta. ALCHEMIA LTD FEBRUARY 2008