JP2002533468A - Inositol-containing hexasaccharide, its synthesis and use - Google Patents

Abstract

  (57) [Summary] The present invention provides compounds of general formula I: Embedded image (Wherein each R is independently hydrogen, an alkyl or substituted alkyl group, an acyl or substituted acyl group, a phosphate group (eg, PO_Three ^-Or PO3H) or a protecting group, or the two R groups may be cyclic phosphate groups; _ThreeOr in the NX group, X represents one or more groups independently selected from hydrogen, alkyl or substituted alkyl, acyl or substituted acyl; NY represents a phthalimido group (NPHt) or N_ThreeOr in NY, Y represents one or more groups independently selected from hydrogen, alkyl or substituted alkyl, acyl or substituted acyl); or a salt of the compound thereof. Or related to the design and synthesis of derivatives. Also disclosed are methods of synthesizing the compounds and their use.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a novel inositol-containing hexasaccharide, in particular, inositol phosphoglycan (
IPG) relates to hexasaccharides that can act as mimetics. It also relates to the synthesis of such hexasaccharides, intermediate compounds formed during their synthesis, the use of hexasaccharides, and compositions containing them.

[0002]

[Prior art]

Many growth factors affect cells by inositol phosphoglycan (IPG)
It is thought to be mediated by a family of second messengers [
1-5]. The source of such IPGs is believed to be the "free" form of glycosyl phosphatidylinositol (GPI) present at the cell membrane. It is believed that IPGs are released by the action of phosphatidylinositol-specific phospholipases after coupling of growth factors to cell surface receptors. IPGs, insulin,
Effects of nerve growth factor, hepatocyte growth factor, insulin-like growth factor I (IGF-I), fibroblast growth factor, transforming growth factor β, IL-2 on B-cells or T-cells, adrenocortical cells ACTH signal, IgE, FSH and hCG stimulation of granulosa cells,
There is evidence that it mediates the action of a number of growth factors, including thyroid stimulating hormone stimulation of thyroid cells, cell proliferation in the ear during early development and in the rat mammary gland.

The family of IPG second messengers can be divided into two different subfamilies, type A and type P, based on biological activity. Type A regulates the activity of many insulin-dependent metabolic effects, such as acetyl-CoA carboxylase (activates), cAMP-dependent protein kinase (inhibits), adenylate cyclase (inhibits), cAMP phosphodiesterase (Stimulate). Conversely, P-type regulates enzyme activity, for example, pyruvate dehydrogenase phosphatase (stimulates) and glycogen synthase phosphatase (stimulates). Type A mimics the lipogenic activity of insulin on adipocytes,
P-type mimics the glycogen-producing activity of insulin on muscle ^[6-9] .

[0004] The soluble IPG fraction has been obtained from various animal tissues, including rat tissues (liver, kidney, muscle, brain, fat and heart) and bovine liver. However, until recently, it was not possible to isolate a single purified component from the tissue-derived IPG fraction because it was less than sufficient to characterize the structure. Thus, previous work was largely based on the biological activity of the fractions, and was based on inferring the identity of the active ingredients based on indirect evidence from metabolic labeling and cleavage techniques.

[0005] In WO98 / 11116 and WO98 / 11117, we describe the isolation of the active components of the A and P type IPG fractions from human liver and placental tissues. The biological activity of these isolates has been identified and certain aspects (eg, mass spectral data) and properties of the structure have been disclosed. Type A substances are, for example,
P-type substances are defined as cyclitol-containing carbohydrates which also contain Zn ²⁺ ions and optionally also phosphoric acid; P-type substances also contain Mn ²⁺ ions and / or Zn ²⁺ ions and optionally also phosphate. It is said to be carbohydrate.

[0006] Other studies have shown that type A IPGs consist of myo-inositol, non-acetylated D-glucosamine, D-galactose and phosphate ^[6] , and that P-type is c-type.
hiro-inositol, non-acetylated D-galactosamine, D-mannose and phosphoric acid ^[7] . We also obtained from a large amount of bovine liver a partially purified glycolipid fraction which, after treatment with bacterial phosphatidylinositol-specific phospholipase C, gave a water-soluble fraction which inhibited cAMP-dependent protein kinase. ^[10] This bioactive agent was partially sequenced and the results were myo-inositol, unacetylated D-galactose, undetermined hexose (D-mannose or D-galactose), and terminal N-acetyl-D- This indicated the existence of a family of substances containing glucosamine residues. In addition, there appeared to be up to four α-D-galactopyranosyl units and up to three phosphate groups ^[10] .

[0007] These partial data have contributed in part to the determination of the chemical structure of type A IPGs, but still leave considerable uncertainty. Nevertheless, it would be desirable to synthesize an IPG analog having an activity that at least partially mimics the activity of a naturally occurring material. For this purpose, synthesis, as described in reference ^[13-17] ,
Structural and biological studies were performed, resulting in the synthesis of many basic substructures, studying their shape and spectroscopic properties, and examining their potential biological activities. For example, we consider the compound C3 (1-D-6-O- (2-amino-2-deoxy-α-D
-Glucopyranosyl) -myo-inositol 1,2- (cyclic phosphate)) and C4 (1-D-6-O- (2-amino-2-deoxy-α-D-glucopyranosyl) -chiro-inositol 1-phosphate) An inositol-containing disaccharide, referred to as ^[14,15] , was synthesized and showed biological activity, at least for myo-inositol-containing C3, in the form of a proliferative effect on the early developing inner ear of chick embryos ^[14] .

[0008] Frick et al. ^[48] also include mannose, glucosamine, and inositol units,
It discloses the synthesis of both trisaccharide and hexasaccharide IPG analogs. They conclude that mannose side chains are required to maximize the insulin mimic activity of their product.

[0009]

Problems to be Solved by the Invention and Means for Solving the Problems

 The present invention provides compounds of the general formula I:

[0010]

Embedded image

Wherein each R is independently selected from hydrogen, an alkyl or substituted alkyl group, an acyl or substituted acyl group, a phosphate group or a protecting group, or two R The group may be a cyclic phosphate group; the NX group represents N ₃ ; or in the NX group X is independently selected from hydrogen, alkyl or substituted alkyl, acyl or substituted acyl.
One represents the above groups; -NY represents a phthalimido group (NPHt) or N _3; or in NY, Y is selected from hydrogen, alkyl or substituted alkyl, independently acyl or substituted acyl Represents one or more groups, provided that the number of phosphate groups in the molecule is 3 or less), or a compound or a salt or derivative of the compound, for designing and synthesizing a novel hexasaccharide. is there.

For example, in the compound of formula I, the NX and NY groups are phthalimide groups (NPht
) Or N ₃ , secondary amine (—NR ₁ H), tertiary amine (—NR ₁ R ₂ ) or quaternary amine
A secondary amine group (—N ⁺ R ₁ R ₂ R ₃ ), an amidino group (R ₁ CONR ₂ —), and X or Y is an acyl group or a substituted acyl group. In these equations,
R ₁ , R ₂ , R ₃ in the NX or NY group are independently selected from hydrogen or C ₁ -C ₁₀ alkyl or substituted alkyl, more preferably C ₁ -C ₅ alkyl. Preferred substituents are phthalimido groups (NPht), N ₃ , NH ₃ ⁺ and AcH
N.

Preferably, the compounds of the formula I contain not more than 3 phosphate groups.

The constituent saccharide units in I are designated af, and in the following description the R groups are referred to according to their position within each unit. For example, R ^2a is the position 2 in the unit a.
Represents an R group.

Compound I is composed of the basic saccharide units considered to be present in type A IPGs, ie, myo-inositol units (a), non-acetylated D-glucosamine units (b), mannose units (c), terminal N -Acetyl-D-glucosamine residue (d) and D-
It contains galactose units (e) and (f). It also includes a rational structural overlap with the conserved linear glycan chain of the GPI anchor described as Formula 2 in FIG. Therefore, the antibody probe generated against 2 is capable of generating I from rat liver.
In view of the immunological evidence ^[11,12] that cross-reacts with PGs and somewhat inhibits the effects of insulin, Compound I is useful as a synthetic analog of naturally occurring A-type IPGs or as a mimetic. And can therefore be reasonably expected to be applicable to pharmaceutical compositions and methods.

Another aspect of the present invention relates to a method of synthesizing a compound of formula I and intermediate compounds formed during the synthesis.

[0017]

BEST MODE FOR CARRYING OUT THE INVENTION

(Compound of Formula I) A first aspect of the present invention provides a compound of Formula I as described above, or a salt or other derivative thereof.

In Formula I, R is an alkyl or acyl group, or a substituted version thereof (eg, a halogeno, cyano, amino, carbonyl, and / or carboxy group, etc.), but preferably C ₁ -C ₁₀ , More preferably C ₁ -C ₅ ,
Grade, second grade, or third grade may be used.

The NX and NY groups are a phthalimide group (NPht) or N ₃ , a secondary amine (—N
R ₁ H), tertiary amine (—NR ₁ R ₂ ) or quaternary amine group (—N ⁺ R ₁ R ₂ R ₃ )
, An amidino group (R ₁ CONR ₂ —), wherein X or Y is an acyl or substituted acyl group. In these formulas, R ₁ , R ₂ , R ₃ in the NX or NY group are independently selected from hydrogen or C ₁ -C ₁₀ alkyl or substituted alkyl, more preferably C ₁ -C ₅ alkyl. Is done.

As used herein, “salts” include physiologically acceptable salts, which means non-toxic salts for pharmaceutically applications. Such salts include those with acidic groups, such as carbonyls, with alkali and alkaline earth metals such as Na, K, Mg, and Ca, and physiologically acceptable organic amines, such as triethylamine and tris (2-hydroxy (Ethyl) amine. Compounds of formula I containing a basic group such as an amino group or a guanidino group (especially the NX or NY group, X and / or Y being hydrogen) are inorganic acids such as hydrochloric acid, sulfuric acid, acetic acid, citric acid, benzoic acid,
Forms salts with maleic acid, fumaric acid, tartaric acid, p-toluenesulfuric acid. Equal numbers of acidic and basic compounds can form internal salts and do not require a third salt component.

“Derivatives” include coordination complexes with metal ions such as, for example, Zn ²⁺ . It also includes the so-called "prodrug" forms of the compounds, which are converted in vitro or in vivo to the compounds of the formula I. Examples of suitable prodrugs are those in which the prodrug is a glycolipid derivative that can be converted to I after phospholipase cleavage and R ^1a is O ‖ -PO-diacylglycerol ‖O.

As mentioned above, the compounds of the present invention may contain one or more groups for protecting hydroxyl or phosphate groups. Phenyl, benzyl, hydroxypropyl nitrile (Hoeben-Weyl, Methods of Organic Chemistry, December 1 or 12/2, Teil
phosphate protecting groups such as heimer, Synthetic Methods of Organic Chemistry, vol. 45).
OH protecting groups for sugars including benzyl, acetyl, benzoyl, pivaloyl, trityl, tert-butyldimethylsilyl, benzylidene or isopropylidene.

A second aspect of the present invention provides a method for chemically or physically binding to a binding partner such as a label, a support substrate, a carrier, an effector or inhibitor molecule or a solidifying agent.
A substance (compound or composition) comprising a compound of Formula I is provided.

In I, a suitable protecting group for R is menthoxycarbonyl (MntCO
), Acetal (in particular, the two R groups together represent a bridged acetal such as O-cyclohexylidene, O-isopropylidene or O-benzylidene), te
rt-butyldimethylsilyl (TBDMS), benzyl (Bn), tert-butyldiphenylsilyl (TBDPS), and the like. The reaction of saccharides with many protecting groups suitable for use in synthesis is well known and described in the standard literature. The choice is, in part,
It will depend on the route by which the compound is synthesized and / or its use, and possibly also on the reactions that it will undergo.

Either, preferably both of R ^1a and R ^6f are phosphoric acid. R ^1a and R ^2a may together represent a cyclic phosphate group. X is preferably H ₃ ⁺ . Y is preferably AcH.

A preferred form of compound I is one that is at least partially deprotected, ie one or more of the R groups is hydrogen. For example, at least in units a and b,
More preferably, the R group in units a, b, c, most preferably in units a, b, c and d, is either a hydrogen or a phosphate group (eg, R ^1a is preferably Acid).

A particularly preferred form of Compound I has been shown in equation 1 in Figure 1, except that the R ^1a and R ^{6 f} Hagarin acid groups are all R groups are hydrogen, X is H ₃ ⁺ and Y are AcH.

Preferred protected forms of I are those prepared or could be prepared by the synthetic methods also provided by the invention (see below). These methods are
In I, there are certain restrictions on the nature of the protecting group R in order to ensure the correct stereochemistry of the glycosidic bond: (a) R ^2c , R ^2e and R ^2f are preferably fixed, non- (B) R ^1a and R ^6f are preferably temporary protecting groups chosen to allow orthogonal deprotection for all fixed protecting groups in I.

Further, NX is preferably a non-participating group, while NY is preferably involved.

The participating groups are those that are involved in the glycosylation reaction and affect the stereochemistry of the formed glycosidic bond, resulting in a 1,2-trans-bond. Non-participating groups are
In principle, they do not affect the stereochemical result of the glycosyl reaction.

Preferred protecting groups for I include X for N ₂ , Y for Pht, R ^2a and R ^3a , R ^4a and R ^5a , R ^4d and R ^6d , R ^3e and R ^4e , and R ^3f and R 3 Crosslinked acetal is included for pair ^4f .

A particularly preferred protected form of I is R^1aIs MntCO; R^2aAnd R^3aIs R^4a
And R^5aR together represent O-cyclohexylidene;^3b, R^6b, R ^2c , R^4c, R^3d, R^2eAnd R^2fIs benzyl (Bn); R^4dAnd R^6dIs O-
Represents benzylidene; R^3eAnd R^4e, And R^3fAnd R^4fIs O-isopropyl
Represents den; R^6fIs acetate.

In some embodiments, the compounds and derivatives of the present invention are, as IPG agonists, more preferably, Type A I, which share one or more biological properties with Type A IPGs.
It is useful as a PG agonist. In other embodiments, the compounds and derivatives of the present invention act as IPG antagonists, more preferably, as Type A IPG antagonists that compete with IPGs and decrease one or more IPG biological activities.

Type A mediators regulate the activity of many insulin-dependent enzymes, such as cAMP-dependent protein kinase (inhibits), adenylate cyclase (inhibits), and cAMP phosphodiesterase (stimulates). Conversely, P-type mediators regulate the activity of insulin-dependent enzymes such as pyruvate dehydrogenase phosphatase (stimulate) and glycogen synthase phosphatase (stimulate). A-type mediators mimic the lipogenic activity of insulin on adipocytes, whereas P-type mediators mimic the glycogen-producing activity of insulin on muscle. Both type A and P type mediators are mitogens when added to fibroblasts in a serum-free medium. The activity of the mediator to stimulate fibroblast proliferation is enhanced when the cells are transfected with the EGF-receptor. Type A mediators can stimulate cell proliferation in chick cochlear vestibular ganglia.

[0035] Soluble IPG fractions having A-type and P-type activities are obtained from various animal tissues including rat tissues (liver, kidney, muscle, brain, adipocytes, heart) and bovine liver. A
Type and P-type IPG bioactivity was detected in human liver and placenta, malaria parasitized RBCs and mycobacteria. Activity of anti-inositol glycan antibodies to inhibit insulin action on human placental trophoblast cells and BC3H1, or on bovine IPG action on rat diaphragm and chick vestibular ganglia, suggests species-spanning conservation of many structural features. . However, despite the prior art including these reports on A-type and P-type IPG activity in some biological fractions, no purification or characterization of the agents involved in that activity is disclosed. It is important to note that.

[0036] Type A substances are cyclitol-containing carbohydrates, which contain Zn ²⁺ ions and optionally phosphate, have the properties of controlling lipogenic activity and inhibiting cAMP-dependent protein kinase. They inhibit adenylate cyclase, and in serum free media,
It is a mitogen when added to GF-transfected fibroblasts and can also stimulate adipogenesis in adipocytes.

P-type substances are cyclitol-containing carbohydrates, which contain Mn ²⁺ and / or Zn ²⁺ ions and optionally phosphate, have the properties of controlling glycogen production and metabolism and activating pyruvate dehydrogenase phosphatase. They also activate glycogen synthase phosphatase, are mitogens when added to fibroblasts in a serum-free medium, and can also stimulate pyruvate dehydrogenase phosphatase.

Methods for obtaining A-type and P-type IPGs are described in WO98 / 11116 and WO9
8/1117.

(Pharmaceutical composition) The third aspect of the present invention relates to a composition comprising the compound, derivative or substance according to the first or second aspect (IPG agonist or antagonist), or a pharmaceutically acceptable derivative thereof. provide.

Pharmaceutical compositions may contain other pharmaceutically acceptable adjuvants, such as carriers, depending on the purpose of the composition and its intended route of administration (eg, oral, intravenous or otherwise).
Buffers, stabilizers or other excipients can be included. They can further include other pharmaceutically active ingredients that can be therapeutically (including prophylactically) active or have some diagnostic function. For example, insulin, P
Type IPG or IPG analog, other type A IPG or analog, and / or IPG
An antagonist may be included. The composition may of course also comprise more than one compound, derivative or substance according to the first or second aspect of the invention.

The composition may be in any suitable form, and may be in the form of tablets, capsules, powders or liquids for oral administration, or solutions or dispersions for use as eg vaccines. Conventional solid or liquid carriers can be used in such preparations. The concentration of the compound, derivative or substance contained in the pharmaceutical composition will, of course, depend on the nature and severity of the condition being treated or diagnosed with the composition, and the patient and method of administration.

The potential uses (including both therapeutic and diagnostic uses) of the pharmaceutical composition of this third aspect of the invention are described below.

Use of Compounds and Compositions In a preferred embodiment, the compounds of formula I are at least partially
It is expected to mimic the biological activity of PGs, and they are also expected to be used in therapeutic and diagnostic methods based on their activity. Thus, the fourth to sixth aspects of the present invention
Are each a compound or derivative according to the first aspect, or a substance according to the second aspect, or an antagonist thereof, for use in a method of surgery, treatment or diagnosis; of such a compound, derivative, substance or antagonist, Use in the manufacture of a therapeutic for use in a method of surgery, treatment or diagnosis; a method of surgery, treatment or diagnosis comprising the use of such a compound, derivative, substance or antagonist.

The term “treatment” as used herein includes prophylaxis. Furthermore, in this section "compound" is understood to include the derivatives, substances and antagonists referred to in connection with the third aspect of the invention.

The compounds are particularly useful for treating and / or treating conditions associated with insulin activity, particularly IPG second messengers (ie, in any way that directly or indirectly causes or is mediated, or is likely to be, related). Or it may be used for diagnosis. They can be used, for example, in the treatment or diagnosis of disorders in which a patient's lipogenic response is somewhat affected and produces relatively low amounts of type A IPGs against growth factors such as insulin.

More specifically, the compounds are insulin resistant, such as diabetes including insulin resistance, insulin resistance in type I diabetes and unstable type diabetes, and insulin resistance such as neurotrophic disorders or polycystic ovarian disease. It may be used in the treatment and / or diagnosis of sexual or insulin hypo-productivity related conditions.

The use of both P-type and A-type IPGs in the diagnosis and treatment of diabetes is described in WO9
8/11435. This application discloses that in some forms of diabetes, the ratio of P: A type IPGs is imbalanced and can be corrected by administration of a medicament comprising an appropriate ratio of P or A type IPGS or an antagonist thereof. It discloses that it is possible. In particular, treatment of obese type II diabetic (NIDDM) patients with P-type IPG and / or A-type IPG antagonists and a mixture of P-type and A-type IPGs, typically about 6: 1 for men and 4: It describes the treatment of IDDM or lean type II diabetes (body mass index <27) with a P: A ratio of 1. The compounds and compositions of the present invention can be used for such types of treatment.

It is contemplated that the compounds of the invention can be used in promoting neuronal proliferation in vitro or in vivo. Thus, they are applicable for the treatment and / or diagnosis of any condition associated with neuronal proliferation. Neurons may be central (brain and spinal cord) neurons, peripheral (sympathetic, parasympathetic, sensory and intestinal) neurons, or motor neurons. Treatment can include treating nervous system damage, motoneuron disease, neurodegenerative disorders or neuropathy. Nervous system loss includes the consequences of trauma, stroke, surgery, infection (eg, viral agents), ischemia, metabolic disease, toxicants, or a combination of these or similar causes. Motor neuron disease includes conditions including spinal muscular atrophy, paralysis or amyotrophic lateral sclerosis. Neurodegenerative disorders include conditions including Parkinson's disease, Alzheimer's disease, epilepsy, multiple sclerosis, Huntington's chorea or Meniere's disease.

Therapeutic treatments in which the compounds can be used include, but are not limited to,
It involves the administration of a therapeutically effective amount of one compound, preferably in the form of a pharmaceutical composition according to the third aspect of the present invention. By “effective amount” is meant an amount effective to provide benefit to the patient (which may be prophylactic), or at least to effect a change in the patient's condition. The amount actually administered to the patient, and the rate and time of administration, will depend on the nature of the patient,
It will depend on the nature and severity of the condition, the manner of administration used, and the like. The trained medical practitioner can select an appropriate value. The compounds can be administered alone or in combination with other treatments, either simultaneously or sequentially. Administration can be by any suitable route, including oral, intravenous, dermal, subcutaneous, parenteral, nasal, intramuscular, intraperitoneal. Methods of targeting directly to an appropriate site, or to a particular site, such as certain cells, can be administered-suitable targeting methods are well known.

The diagnostic method according to the invention may be used to determine, qualitatively or quantitatively, a compound (including, of course, an antagonist), or its identification, for determining the presence or alteration of a particular medical condition or condition. Could be used, or of a species that competes in binding with other specific binding partners. Such methods can be performed in vitro or in vivo. One or more substances used in the method can be appropriately labeled.

[0051] A seventh aspect of the invention comprises combining one or more compounds with one or more pharmaceutically acceptable adjuvants and / or one or more other therapeutically active agents. Methods for preparing pharmaceutical compositions are provided.

Compound Synthesis A further aspect of the invention relates to the synthesis of compounds of formula I. A preferred strategy is to build three intermediate disaccharide "building blocks" from four already available monosaccharide units.
To enable their preparation. These disaccharide intermediates have the general formulas II, III,
And IV.

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Embedded image Wherein R, X, and Y are as defined in Formula I, and each L is a leaving group that activates the anomeric position of the corresponding sugar unit in the preparation of a glycosylation reaction at a glycosyl acceptor. (Or “activating group”), or a leaving group precursor (ie, a group that can be converted to a suitable leaving group). Each L depends on the response to the unit in question:
It must be chosen to ensure an optimal balance between reactivity at the anomeric position and selectivity in units as a whole.

The constituent saccharide units of the components of compounds II-IV are again labeled af, reflecting their corresponding units in formula I.

In compound II, position 1 of unit a is ideally distinguished from other positions of the unit by the selection of an appropriate R group. For example, R ^1a may be MntCO and the remainder of the unit is protected as dicyclohexylidene acetal, where R ^2a and R ^3a , and separately R ^4a and R ^5a , together represent the bridging group O-cyclohexylidene. You may. X
Is preferably N ₂ . R ^4b is conveniently H, or a removable precursor group such as TBDMS, for subsequent preparation of glycosylation of II. R ^3b and R ^6b are Bn
And the like.

In compound III, L is preferably trichloroacetimidate (C (NH
) CCl_Three) Or a leaving group precursor such as thiophenyl (SPh). SPh
Preferred elimination because of its stability and convertibility that can be easily converted to many suitable leaving groups
It is a base precursor. R^2cMust be fixed and non-participating groups, preferably
Is Bn. R^3cAnd R^4cSuitable groups of include Bn and TBDPS, preferably R ^4c Is Bn, R^3cIs TBDPS. R^3cIs preferably R^2cAnd R^4cBe different from
It is. In the unit d, NY must be a participating group, preferably NPh
t and a suitable protecting group R is benzylidene acetal for positions 4 and 6,
Position 3 contains an acetal bridging group such as Bn.

In compound IV, L is preferably a leaving group precursor such as trichloroacetimidate or thiophenyl. Positions 2, 3, and 4 of both units and unit f
The R group at position 6 of is a suitable protecting group such as Bn, Ac, or (ideally for positions 3 and 4 of each unit) a bridged acetal such as O-isopropylidene. However, R ^2e and R ^2f must be fixed, non-participating protecting groups, such as Bn, and R ^6f has orthogonal deprotection for all fixed protecting groups in the final product I. It must be a temporary protecting group chosen to enable it.

Thus, an eighth aspect of the present invention provides a method of synthesizing a compound of formula I, comprising the use, preferably a preparation, of at least two intermediate disaccharide compounds II, III and IV. This method conveniently involves condensing at least two disaccharides together to form a tetrasaccharide intermediate compound. The hexasaccharide product is preferably synthesized by reacting the tetrasaccharide intermediate with a third disaccharide intermediate, also preferably selected from compounds II, III and IV. The method preferably comprises the step of reacting at least compounds II and III, more preferably all three compounds II, III and IV together.

The method optionally involves the removal of one or more protecting groups R after formation of the hexasaccharide and / or replacement with another group such as, for example, phosphoric acid to produce compound 1. Conventional chemical techniques can be used to effect such substituent transformations, the nature and continuity of the reaction steps employed depending on the nature of the R group and the group being substituted.

Preferably, compounds II and III are first reacted together to form an intermediate tetrasaccharide of general formula XI:

[0060]

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Wherein R, X, and Y are as defined for compounds II and III, and R ^3c is preferably hydrogen.

Compound XI is then preferably glycosylated using compound IV as a glycosyl donor to form compound I. This last step will involve the selective deprotection of compound XI at R ^3c to convert to a glycosyl acceptor.

A ninth aspect of the present invention therefore provides a method of synthesizing a compound of formula I, comprising the use of a tetrasaccharide intermediate of formula XI and preferably the intermediate of said intermediate with a compound of formula IV The reaction preferably also includes a preparation.

Compounds II, III and IV are preferably prepared from monosaccharide “building blocks” of the general formula V-IX.

[0065]

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[0066]

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[0067]

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[0068]

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[0069]

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Wherein R, X, Y and L are as defined for Formula II-IV, and the units having letters also correspond to those given for Compound I-IV (Compound IX is unit
provide both e and f)]

Compound II is preferably prepared by first preparing myo-inositol building block V and then glycosylating using a D-glucosamine derivative such as VI as a glycosyl donor. In the myo-inositol building block, position 6 should be free for glycosylation (
That is, R ^6a should be hydrogen), position 1 should be distinguished, ideally as described for compound II. Compound V is derived from myo-inositol
It can be prepared using a regioselective acylation reaction via a boron-tin exchange reaction ^[25-30] .

The glycosyl donor is preferably 2-azido-2-deoxy-D-glucopyranosyl, ie compound VI where X = N ₂ . L is preferably a trichloroacetimidate leaving group, which can be a precursor during synthesis, for example with a thiophenyl group. Such compounds can be prepared, for example, from 2-amino-2-deoxy D-glucosamine hydrochloride via a diazo transfer reaction from methanesulfonyl azide trifluoride ^[37] .

R ^3b , R ^4b , and R ^6b in the glycosyl donor can be any suitable protecting group, for example, R ^3b and R ^6b are Bn and R ^4b is TBDMS (optionally removable). . Generally, R ^4b should differ from R ^3b and R ^6b, and more preferably R ^3b and R ^6b are the same. Cross-linked acetal is not preferred as a protecting group because subsequent reductive ring opening during the glycosylation reaction can result in hydrolysis of the acetal protecting group present on the myo-inositol unit. In compound II, R ^4b should ultimately be H, position 4b should be selectively deprotected after glycosylation of the myo-inositol block.

Compound III is preferably prepared by glycosylation of mannose derivative VII with glucosamine derivative VIII. In VII, L is ideally a leaving group precursor such as thiophenyl, which after glycosylation at VIII can be converted to a suitable leaving group. R ^6c is ultimately hydrogen, leaving that position free for glycosylation. Suitable groups for R ^2c , R ^3c and R ^4c include Bn and TBDP
S is included, and preferably, R ^2c and R ^4c are Bn, and R ^3c is TBDPS. R ^2c is
It must be a fixed non-participating group as in III. R ^3c and R ^6c are mutually orthogonal temporary protecting groups chosen to allow deprotection without affecting the residual groups at positions 1 and 4.

In the glucosamine derivative VIII used as a glycosyl donor, position 3
, 4, and 6 must be protected, but L must be a suitable leaving group, preferably fluoride. Suitable protecting groups include acetal bridging groups such as benzylidene acetal for positions 4 and 6, and Bn for position 3. In the synthesis of compounds, NY must be a participating group such as NPht.

Compound IV is preferably a glycosyl donor based on D-galactose.
Receptor, (both corresponding to formula IX, both of which are β-D-
(Which can be prepared from a suitable D-galactose derivative such as taacetate)
It is prepared by reacting In compound IX, a glycosyl donor
L is a suitable leaving group such as trichloroacetimidate;^2e, R ^3e , R^4e, And R^6eIs Bn, Ac or (ideally positions 3 and 4) O-isopro
Represents a suitable protecting group such as a bridged acetal such as pyridene; Glycosyl receptor and
Then R^6eShould be hydrogen for easy glycosylation and the position
2, 3 and 4 are Bn or (also position 3 and 4 are convenient) O-isopropylidene etc.
Must be appropriately protected by a cross-linked acetal or the like. L is a leaving group precursor
It must be a body, preferably a thiophenyl group. One about substituents
Is restricted to R at both the glycosyl donor and acceptor.^2eNon-participating group with fixed
And both are preferably Bn.

Thus, the method of the present invention more preferably comprises the four monosaccharide units myo-inositol, D-glucosamine, D-mannose, and β-D-galactose (all of which have, for example, one or more hydroxyl groups). Which may be in the form of a derivative such as pentaacetate, substituted with a suitable protecting group). The preferred four source materials are:

[0078]

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[0079]

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[0080]

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[0081]

Embedded image

Therefore, the tenth aspect of the present invention relates to a myo-inositol, D-glucosamine, D-mannose and β-D-galactose as basic monosaccharide raw materials.
And / or a method of synthesizing a compound of formula I, comprising the use of a suitable derivative thereof. β-D- is preferably lactose itself, instead of β-D-galactopyranose pentaacetate.

Obviously, the protecting groups used during the synthesis must be carefully chosen to ensure that they are always available for a suitable substituent. A suitable group is
Reference can be made to those for compounds II-IX.

The substituents referred to as preferred in compounds V-IX and / or in compounds II-IV are, in corresponding positions, in the tetrasaccharide intermediate XI and in the final product
Naturally preferred in I.

Many intermediate compounds formed during the synthesis according to the invention are novel compounds
it is conceivable that. These are compounds 9, 10, 11, 12, 13, 18, 19, 20
, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 35, 36
, 37, 38, 39, 40, 41, 42, 43, 44, 45, 48, 49, or
50, and are described in the Examples below. In particular, the eleventh aspect of the present invention
There is provided a compound of general formula II, or a salt or derivative thereof, wherein R^4b(Preferably
Selected to allow orthogonal deprotection for hydrogen or other protecting groups present
Protecting group) is R^3bUnlike R^6bAnd more preferably R^3bAnd R ^6b Are the same as each other but R^4bAnd different.

The twelfth to eighteenth aspects of the present invention each have the general formulas III and I as defined above.
It provides compounds of V, VI to IX and XI, or in each case salts or other derivatives.

A nineteenth aspect of the present invention is to provide a glycosyl donor in the form of 2-azido-2-deoxy-D-glucopyranosyl of the formula VI wherein X = N ₂ , Preparation from D-glucosamine salt by a diazotransfer reaction from methanesulfonyl azide trifluoride (shown in FIG. 4) and then the donor is obtained by converting m of the formula V
There is provided a method of synthesizing a compound of formula II (defined above), comprising reacting with a glucosyl receptor in the form of a building block of yo-inositol.
The method preferably also includes the preparation of compound V. The nature of the R and L groups is preferably as described for the preparation of intermediate II. In particular, R ^4b is preferably different from the other R groups of the glycosyl donor, so that in product II position 4b is free for further glycosylation, for example the use of the synthesis of tetrasaccharide XI and / or hexasaccharide I. ing.

Finally, a twentieth aspect of the present invention provides a method for synthesizing a glycosyl acceptor of formula II wherein R ^4b is hydrogen and a glycosyl donor of formula III wherein L is a leaving group, There is provided a method of synthesizing a tetrasaccharide of general formula XI as defined above. The method preferably comprises preparing one or both of compounds II and III, and more preferably comprises preparing compound II according to the nineteenth aspect of the invention.

Hereinafter, embodiments of the present invention will be described in more detail with reference to the drawings, but the present invention is not limited thereto.

[0090]

【Example】

Examples of the synthesis of the compounds of the formula I according to the invention are described. The synthesis was devised using the retrosynthetic analysis described in FIG. It can be prepared per se (via the monosaccharide unit V-IX) from the four raw materials myo-inositol, D-glucosamine, D-mannose and β-D-galactopyranose pentaacetate, II
, III and IV proceeded through three intermediate disaccharide compounds.

The choice of protecting groups and leaving groups for the monosaccharide “building block” V-IX depends on the selective reaction of the appropriate substituents during each step of the synthesis and the desired reaction at each glycosidic bond formed. Made to ensure stereochemistry.

Preparation of Intermediate II FIG. 3-5 illustrates the preparation of disaccharide II from myo-inositol and D-glucosamine. Myo-inositol building block 6 was prepared according to a previously reported method ^[25,26] based on the established regioselective enhancement of the nucleophilicity of hydroxyl groups such as tributyltin ether or dibutyltin acetal ^[27] . Overcomes the insolubility of myo-inositol in many organic solvents using a boron-tin exchange reaction ^[28-30] (For background on the preparation of myo-inositol derivatives, Potter
See also ^[24] .

Thus, myo-inositol was converted to the hexane-soluble hexa-O-diethylboryl derivative 3 (100% yield), which was in situ dibutyltin bis-acetylacetonate and then L-menthyl chloroformate. Reaction yielded a diastereomeric mixture of regioselective monosubstituted derivatives 4 and 5, from which the desired diastereoisomer 4 could be separated. 4 was then transformed into building block 6, and after protection of dicyclohexylidene acetal, etc., position 6 was free for glycosylation and elimination position 1 was different.

The cyclohexylidene acetal of myo-inositol is often a glycosyl
It has been used as an intermediate in the preparation of myo-inositol ^[31,32] . Compound 6 was most conveniently prepared using 1-ethoxycyclohexene for acetalization reactions in cyclohexanone under conditions of thermodynamic control.

The 1,2-cis glycosylation of 6 was conveniently performed using 2-azido-2-deoxy-D-glucopyranosyl trichloroacetimidate as a glycosyl donor. Although 2-azido-2-deoxy-glycosyl donors are currently used in oligosaccharide synthesis, many methods used for the preparation of 2-azido-2-deoxy building blocks have low diastereoselectivity. And many steps ^{[33-36 ]} . However, peracetylation from commercially available 2-amino-2-deoxysugar hydrochloride by diazotransfer reaction from methanesulfonyl azide trifluoride.
A one-pot synthesis of -azido-2-deoxysugar has been reported ^[37] , and this method is used to prepare glycosyl donors 15 and 17 as shown in FIG.

D-glucosamine hydrochloride is, for example, tetra-O-acetylated 2-azido-2-
It is converted to the deoxy derivative 8 ^[37] , which in turn is converted to thioglycoside 9 ^[38] , which is then converted to ^acetimidate trichloride 15 and 15 using established chemical techniques ^{[18,23,39-43].} 17 were transformed via intermediates 10-14, 10, 11, and 16, respectively (see Materials and Methods).

Glycosylation of myo-inositol building block 6 with glycosyl donor 17 in the presence of trimethylsilyl triflate in diethyl ether ^[23] yielded 18 as a 10: 1 α / β mixture in 95% yield ( (Fig. 5). Subsequent reductive ring opening of the benzylidene acetal ^[41] in this mixture, however, resulted in partial hydrolysis of the cyclohexylidene acetal. To prepare intermediate compound II, donor 15 was therefore preferred for glycosylation.

Thus, as shown in FIG. 5, under the above conditions, 15 condensed with 6 to give 19 (corresponding to intermediate II) as a 9: 1 α / β mixture in 73% yield. Treatment of 19 with tetrabutylammonium fluoride ^[44] resulted in product 20 (position 4 of the glucosamine unit was deprotected) in 83% yield.

Preparation of Intermediate III As shown in FIG. 6, compound III was prepared using a readily available mannose derivative, 1,
Prepared from 6-anhydro-β-D-mannopyranose (21) via protected mannose unit 29, then glycosylated with a fluorinated glucosamine derivative. The conversion from 20 to 28 was performed according to the method described by Klooterman et al. ^[36] . Glycosylation of 29 was conveniently performed according to the methodology reported to Suzuki ^[47] to give disaccharide 31 with high yield and selectivity. This was then converted to ^acetimidate trichloride 33 (ie, intermediate III) via compound 32 ^[18,19,23] (see Materials and Methods).

Preparation of Intermediate IV FIG. 7 shows a galactose derivative, β-D-galactopyranose pentaacetate (
3 shows the preparation of IV from 34). This was converted to glycosyl acceptor 35 ^{[21,38,45 ]} , which was further converted to glycosyl donor 38 via 36 and 37 (see Materials and Methods).

The glycosylation reaction of 35 and 38 yielded the disaccharide 39 as a 6: 1 α / β mixture in 86% yield. This further translates to acetimidate trichloride 41, 40
(See Materials and Methods) ^[18,19,23] . 41 corresponds to intermediate IV.

FIG. 8 shows the preparation of the hexasaccharide product 44 corresponding to compound I. First, the disaccharide 20 (
Intermediate II) and 32 (Intermediate III) are combined and condensed to give high stereoselectivity and 8
The tetrasaccharide 42 was obtained with a yield of 1%. Carefully controlled desilylation of 42 results in a high yield of glycosyl acceptor 43 (corresponding to XI), which has a hydroxyl group at position 3 of the D-mannose unit.

[0103] 43 was then glycosylated with acetimidate trichloride 41 (intermediate IV) to give the hexasaccharide 44 as a 6.5: 1 α / β mixture in 83% yield.

Deprotection of 44 To obtain compound 1 from 44, conventional methods for removing each protecting group can be used. For example, R ^1a can be removed first with excess LiOH in THF / MrOH at room temperature. This simultaneously removes the acetate group R ^6a ,
It will also cause partial ring opening of the phthalimide group in unit d. The phthalimide group can be re-closed by treatment with Et ₃ N / Ac ₂ O, which will result in acetylation of both R ^1a and R ^6f . The phthalimide group is followed by
It can be removed, for example, at 90 ° C. in n-butanol using a large excess of ethylenediamine and the resulting amine is then acetylated under normal conditions. O-deacetylation then gives the diol (ie, R ^1a and R ^6f are H), which can be subjected to phosphorylation using the phosphoramidite method. Finally, 10% P
Treatment with hydrogen in the presence of d / C will give the final deprotected product 1.

Materials and Methods General Description: TLC was performed on pre-coated plates (Merck aluminum sheet silica 60
F ₂₅₄ , Art. no. 5554). Detection was performed by observation under UV light (254 nm), followed by visualization by using sulfuric acid or phosphomolybdic acid in EtOH and then heating. Silica Gel using deflash method
60 (0.023-0.040 mm, E. Merck) column chromatography was performed. Richer J
Melting points were measured using an ung Thermovar apparatus and were not corrected. Specific rotations were measured on a Perkin Elmer model 241 polarimeter. NMR spectra were obtained using Bruker AMX-200, A
vance DRX-500, Varian Gemini-200, XL-300 or Unity 500 spectrometer. Chemical shifts are expressed in ppm and reference is made to the residue signal of the solvent used. Hera
us CHNO-Rapid equipment Analysts of Instituto de Quimica Organica General (CSIC)
Micro analysis was performed by is Department.

2,3: 4,5-di-O-cyclohexylidene-1-O-(−)-menthoxycarbonyl-1D-myo-inositol (6) and 2,3: 5,6-di- O-cyclohexylidene-1-O-(-)-menthoxycarbonyl-1-D-myo-inositol (7) 1-O-(-) menthoxycarbonyl-myo-inositol ^[26] (4 )
100 mg (0.276 mmol) and 5.7 mg of dry p-TsOH (0.03 m
mol) in a solution of 350 mL (2.76 mmol) in 2 mL of cyclohexanone.
1) -Ethoxycyclohexene was added. The reaction mixture was stirred for 3 hours and 30 minutes, quenched with Et ₃ N, and evaporated. 73 mg (50%) of 6 and 41 mg (28%) by silica gel column chromatography (hexane-EtOAc, 5: 1).
7 was obtained. Compound 6: white solid. Rf (hexane-EtOAC, 4: 1) = 0.26

[0107]

Compound 7: colorless oil. Rf: 0.14 (hexane / EtOAC 4: 1)

[0109]

Phenyl 3,4,6-tri-O-acetyl-2-azido-2-deoxy-1-
Thio-D-glucopyranoside (9) 2.10 g (5.63 mmol) of CH at room temperature in 1,3,4,6-tetra-O-acetyl-2-azido-2-deoxy-D-glucopyranose ^[37]. ₂ Cl ₂ 4
To the solution in 5 mL was added 1.15 mL (11.25 mmol) of thiophenol and 3,12 mL (25.31 mmol) of boron trifluoride diethyl etherate. The reaction mixture was stirred for 8 days, diluted with CH ₂ Cl ₂ and washed with NaCl,
Dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-E
At tOAC (3: 1), 1.58 g (66%) of 9 was obtained as a 3: 1 α / β mixture, and 0.53 g (25%) of 8 was recovered. Rf (hexane-EtOAC,
3: 1) = 0.27 ¹ H NMR of 9α obtained from the spectrum of the α / β mixture.

[0111]

¹ H NMR of 9β obtained from the spectrum of the α / β mixture

[0113]

Phenyl 2-azido 4,6-O-benzylidene-2-deoxy-1-thio-
D-Glucopyranoside (10) To a solution of 3.00 g (7.09 mmol) of 9 in 110 mL of MeOH at room temperature was added 5 mL (0.3 M) of sodium methoxide in MeOH. After 20 minutes, the solution was neutralized with Amberlite IR-120, filtered and evaporated. The obtained crude mixture of phenyl 2-azido-2-deoxy-1-thio-D-glucopyranoside was
Dissolved in 0 mL CH ₃ CN. Benzaldehyde dimethyl acetal 5.3
2 mL (35.45 mmol) and 67.4 mg (0.3
Was added 5 mmol), the reaction mixture was stirred at room temperature for 2 hours, quenched with Et ₃ N, and evaporated. Silica gel column chromatography (hexane-EtOAC,
6: 1), 1.85 g of 10α and 0.80 g of 10β (7: 3 ratio, total yield 9
7%). Data for 10α: white solid. Rf (Hexane-EtOAC, 3: 1)
= 0.34.

[0115]

Data for 10β: white solid. Rf (Hexane-EtOAC, 3: 1)
= 0.36.

[0117]

Phenyl 2-azido-3-O-benzyl-4,6-O-benzylidene-2-deoxy-1-thio-D-glucopyranoside (11) 546 mg (1.42 mmol) of 10β in 9 mL of DMF at room temperature In a solution of sodium hydride (43 mg, 1.70 mmol) followed by benzyl bromide
. 21 mL (2.84 mmol) were added. The reaction mixture is stirred for 40 minutes,
Quenched with a saturated aqueous solution of aHCO ₃ and dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-EtOAC) provided 11β in 98% yield. According to the same method, 11α was synthesized in a yield of 95% using 10α as a raw material. Data for 11β: white solid. Rf (Hexane-EtOAC, 3: 1)
= 0.58.

[0119]

Data for 11α: white solid. Rf (Hexane-EtOAC, 3: 1)
= 0.54.

[0121]

Phenyl 2-azido-3,6-di-O-benzyl-2-deoxy-1-thio-
D-glucopyranoside (10) 11β 447 mg (0.94 mmol) of THF9 containing 3Å molecular sieve
. The solution in 4 mL was stirred at room temperature for 30 minutes. Thereafter, 1.201 g (18.
16 mmol) of sodium cyanoborohydride was added. A saturated solution of hydrogen chloride in diethyl ether was then added dropwise until the gas had ceased evaporating (pH <7) and TLC analysis indicated conversion of all starting material. The mixture was neutralized with a saturated aqueous solution of NaHCO ₃ , diluted with CH ₂ Cl ₂ , filtered through celite, washed with water,
Dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-E
tOAC (3: 1) provided 430 mg (96%) of 12β as a colorless oil. According to the same method, 12α was synthesized in 87% yield using 11α as a starting material. Data for 12β: Rf (hexane-EtOAc, 3: 1) = 0.28
.

[0123]

Data for 12α: Rf (hexane-EtOAc, 3: 1) = 0.31
.

[0125]

Phenyl 2-azido-3,6-di-O-benzyl-4-O- (tert-butyldimethylsilyl) -2-deoxy-1-thio-D-glucopyranoside (13) 345 mg (0.72 mmol) Of 12β and 287 mL (2.17 mmol)
Of collidine in 1 mL of CH ₂ Cl ₂ was cooled at 0 ° C. 249 mL (1.08 mmol) of tert-butyldimethylsilyl triflate was added dropwise for 2 hours. The reaction mixture was stirred for 10 minutes, quenched with water / ice, diluted with CH ₂ Cl _2, extracted, washed with brine and dried over Na ₂ SO _4. Silica gel column chromatography provided 405 mg (95%) of 13β as a colorless oil. According to the same method, 13α was synthesized in a yield of 98% using 12α as a raw material. Data for 13β: Rf (hexane-EtOAc, 5: 1) = 0.69
.

[0127]

Data for 13α: Rf (hexane-EtOAC, 5: 1) = 0.62
.

[0129]

2-azido-3,6-di-O-benzyl-4- (tert-butyldimethylsilyl) -2-deoxy-D-glucopyranose (14) 331 mg (0.56 mmol) of 13β in 12 mL of acetone The solution was cooled to −15 ° C. in the dark and 129 mg (0.73 mmol) of NBS was added. 4
After 5 minutes, the reaction mixture was quenched with a saturated aqueous solution of NaHCO ₃ , diluted with EtOAc and extracted, washed with brine and dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-EtOAC, 7: 1) provided 279 mg of 14 as an 11: 1 α / β mixture of anomers (quantitative yield). Following the same method,
14 was synthesized in 91% yield using 13α as a starting material. Rf (hexane-EtOAC, 6: 1) = 0.15.

[0131]

2-Azido-3,6-di-O-benzyl-4-O- (tert-butyldimethylsilyl) -2-deoxy-D-glucopyranosyl trichloroacetimidate (
15) To a solution of 241 mg (0.48 mmol) of 14 in 2.5 mL of CH ₂ Cl ₂ at room temperature was added 484 mL (4.83 mmol) of trichloroacetonitrile and 67 mg (0.48 mmol) of flame dried potassium carbonate. After 1 hour and 45 minutes, the reaction mixture was diluted with CH ₂ Cl ₂ , filtered through celite and evaporated under reduced pressure.
By silica gel column chromatography (hexane-EtOAC, 10: 1),
198 mg of 15α and 85 mg of 15β (7: 3 ratio, 91% overall yield) were obtained. Data for 15β: Rf (hexane-EtOAc, 6: 1) = 0.43
.

[0133]

Data for 15α: white solid. Rf (hexane-EtOAC, 6: 1)
= 0.38.

[0135]

2-Azido-3-O-benzyl-4,6-O-benzylidene-2-deoxy-D
-Glucopyranose (16) To a solution of 80 mg (0.168 mmol) of 11α in 1.7 mL of acetone, cooled to -15 ° C in the dark, was added 51.5 mg (0.289 mmol) of NBS. After stirring for 1 hour and 15 minutes, the reaction mixture was quenched with a saturated aqueous solution of NaHCO ₃ , diluted and extracted with EtOAc, washed with brine and dried over Na ₂ SO ₄ .
Silica gel column chromatography (hexane-EtOAc, 2: 1) gave 6
2 mg (96%) of 16 was obtained as a mixture of white solid, α / β (1: 1) isomer. Rf (hexane-EtOAC, 2: 1) = 0.41.

[0137]

2-Azido-3-O-benzyl-4,6-O-benzylidene-2-deoxy-D
- glucopyranosyl trichloromethyl acetimidate (17) at room temperature, to a solution in CH ₂ Cl ₂ 2.5mL of 16 of 177 mg (0.46 mmol), trichloro acetonitrile 463ML (4.62 mmol) and activated potassium carbonate 64 mg (0. 46 mmol) was added. After 1 hour 30 minutes, the reaction mixture was diluted with CH ₂ Cl ₂ , filtered through celite and evaporated. 99 mg of 17β was obtained by silica gel column chromatography (hexane-EtOAc, 3: 1).
And 125 mg of a 1: 5 mixture of 17α and 17β (92% overall yield, 1:10
α / β mixture). Data for 17β: Rf (hexane-EtOAc, 4: 1) = 0.45
.

[0139]

Data for 17α: Rf (hexane-EtOAc, 4: 1) = 0.36
.

[0141]

6-O- [2-azido-3-O-benzyl-4,6-O-benzylidene-2-deoxy-α-D-glucopyranosyl] -2,3: 4,5-di-O-cyclohex Siliden-1-O-menthoxycarbonyl-1D-myo-inositol (18α
) 56 mg (0.11 mmol) of 17β, 23 mg (0.04 mmol) of 6
And a mixture of powdered 4 molecular sieves in 1.1 mL of ethyl ether
Stirred at room temperature for 5 minutes. At this time, 76 mL (0.008 mmol) of a solution of trimethylsilyl triflate in ethyl ether (0.108 M) was added dropwise. The reaction mixture was stirred for 15 minutes, quenched with triethylamine, diluted with CH ₂ Cl _2,
Filtered over celite and evaporated on vacuo. Silica gel column chromatography (hexane-EtOAC, 12: 1) showed that 18 was a 10: 1 anomeric α /
Obtained as a β mixture (37 mg, 95%). Data for 18α: Rf (hexane-EtOAc, 6: 1) = 0.38
.

[0143]

6-O- [2-azido-3,6-di-O-benzyl-4-O- (tert-butyldimethylsilyl) -2-deoxy-α-D-glucopyranosyl] -2,3: 4
, 5-Di-O-cyclohexylidene-1-O-menthoxycarbonyl-1D-m
A mixture of 180 mg (0.28 mmol) of 15β, 73 mg (0.14 mmol) of 6, and powdered 4 molecular sieve in 3 mL of ethyl ether was added to 45 mg of yo-inositol (19α).
Stirred at room temperature for minutes. At this time, 194 mL (0.02 mmol) of a solution of trimethylsilyl triflate in ethyl ether (0.108 M) was added dropwise over 45 minutes. The reaction mixture was stirred for 15 minutes, quenched with triethylamine, CH ₂ C
diluted with l _2, filtered through Celite, and evaporated in vacuo. Silica gel column chromatography (hexane-EtOAC, 12: 1) afforded 19 as a 9: 1 anomer α / β mixture (102 mg, 73%). Data for 19α: Rf (hexane-EtOAc, 3: 1) = 0.76
.

[0145]

6-O- (2-azido-3,6-di-O-benzyl-2-deoxy-α-D-glucopyranosyl) -2,3: 4,5-di-O-cyclohexylidene-1 -O-menthoxycarbonyl-1D-myo-inositol (20α) To a solution of 82 mg (0.08 mmol) of 19α in 0.8 mL of THF,
204 mL of a 1 M solution of tetrabutylammonium fluoride in THF (0.24 mm
ol) was added. The reaction mixture was stirred for 45 minutes, quenched with water, diluted with CH ₂ Cl ₂ , extracted and washed with brine. Silica gel column chromatography (hexane-EtOAC, 6: 1) provided 20α (61 mg, 84%). Rf (hexane-EtOAC, 3: 1) = 0.46.

[0147]

1,6-Anhydro-3-O- (4-methoxybenzyl) -β-D-mannopira
North (23) 1,6-anhydro-2,3-O-endo- (4-methoxybenzyl) at 0 ° C.
Den) -β-D-mannopyranose^[36]3.5 g (12.5 mmol) of CH _Two Cl_Two To a solution in 125 mL was added a solution of DIBALH in toluene (1 M).
0 mL (40 mmol) was added slowly. After 5 hours, Et_ThreeN and MeOH
Was added. Dilute the crude reaction mixture with EtOAc and wash with a solution of HCl (10%)
And extracted with EtOAc. Evaporate the organic layer and remove the crude with a silica gel column.
Chromatography (CH_TwoCl_Two-MeOH, 20: 1) and 2.8 g of
23 was obtained (79%). Rf (CH_TwoCl_Two-MeOH, 20: 1) = 0.16.

[0149]

1,6-Anhydro-2,4-di-O-benzyl-3-O- (4-methoxybenzyl) -β-D-mannopyranose (24) 2.4 g of 23 at room temperature (8 .5 mmol) in 20 mL of DMF.
472 mg (18.7 mmol) of NaH and 1.9 mL of BnBr (25.5 mmol)
l) was added. After 2 hours, it was added MeOH, and the reaction mixture was diluted with EtOAc, washed with H ₂ O, dried over Na ₂ SO _4, and evaporated. By silica gel column chromatography (hexane / EtOAC, 3: 1), 3.9 g of 24 (quantitative yield) Rf (hexane-EtOAC, 2: 1) = 0.31.

[0151]

4.88 g of 1,6-di-O-acetyl-2,4-di-O-benzyl-3-O- (4-methoxybenzyl) -α-D-mannopyranose (25) 24 ( A solution of 10.55 mmol) and 240 mL (1.24 mmol) of trimethylsilyl trifluoromethanesulfonate in 33 mL of acetic anhydride was stirred at 0 ° C. for 1 hour and at room temperature for 2 hours. The reaction mixture was diluted with EtOAc and N
Washed carefully with a saturated aqueous solution of aHCO ₃ , extracted with EtOAc and dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-EtOAC, 3
1) yielded 25α (4.72 g, 79%) and 24β (169 mg, 3%). Data for 24α: Rf (hexane-EtOAC, 2: 1) = 0.36
.

[0153]

Data for 24β: Rf (hexane-EtOAC, 2: 1) = 0.31
.

[0155]

1,6-Di-O-acetyl-2,4-di-O-benzyl-α-D-mannopyranose (26) 100 mg (0.18 mmol) of 25 in 1.5 mL of CH ₂ Cl ₂ To a solution of was added 50 mL of trifluoroacetic acid in ₂ mL of CH ₂ Cl ₂ . The reaction mixture was stirred at room temperature for 3 hours, neutralized with a saturated aqueous solution of NaHCO ₃ , extracted with CH ₂ Cl ₂ and dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-
EtOAC (3: 1) gave 26 (77 mg, 98%). Rf (hexane-
EtOAC (2: 1) = 0.24.

[0157]

The 1,6-di-O-acetyl-2,4-di-O-benzyl-3-O- (tert-
1.40 g (3.15 mmol) of butyldiphenylsilyl) -α-D-mannopyranose (27) 26, 4-dimethylaminopyridine 170
mg (1.39 mmol), 857 mg (12.60 mmol) of imidazole in 5 mL of DMF were added to 1.64 mL (6.30 mmol) of tert-chloride.
Butyldiphenylsilyl was added. The reaction mixture was stirred at room temperature for 17 hours, diluted with ethyl ether, washed with water and brine and dried over Na ₂ SO _4. Silica gel column chromatography (hexane-EtOAC, 10: 1, 4: 1) provided 27 (1.91 g. 89%). Rf (hexane-EtOAC, 2: 1)
= 0.55.

[0159]

Phenyl 6-O-acetyl-2,4-di-O-benzyl-3-O- (tert
-Butyldiphenylsilyl) -1-thioα-D-mannopyranoside (28a) To a solution of 1.80 g (2.64 mmol) of 27 in 26 mL of CH ₂ Cl ₂ at room temperature, 592 mL (4.80 mmol) of thiophenol And boron trifluoride diethyl etherate (1.32 mL, 10.5 mmol) were added. The reaction mixture was stirred for 30 minutes, quenched with a saturated aqueous solution of NaHCO ₃ and the organic layer was washed with Na ₂ S
Dried over O ₄ . Silica gel column chromatography (hexane-EtOA
C, 10: 1) to give 28α (1.735 g) and 28β (115 mg) in an overall yield of 9
7% (15: 1 α / β ratio). Data for 28α: Rf (hexane-EtOAc, 5: 1) = 0.40
.

[0161]

Phenyl 2,4-di-O-benzyl-3-O- (tert-butyldiphenylsilyl) -1-thio-α-D-mannopyranoside (29) 100 mg (0.14 mmol) of 28α in 2 mL of methanol Into the solution
0.4 mL of sodium methoxide (1M) in methanol was added. The reaction mixture was stirred at room temperature for 1 hour, neutralized with Amberlite IR-120 H ⁺ , filtered and evaporated. By silica gel column chromatography (hexane-EtOAc, 3: 1)
29 (95 mg, quantitative yield) was obtained. Rf (Hexane-EtOAC, 3: 1)
= 0.46.

[0163]

3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-β-D-glucopyranosyl fluoride (30) 100 mg (0%) of -15C in CH ₂ Cl _2. .17 mmol) of 3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-1-thio-
68 mL (0.52 mmol) of diethylaminosulfur trifluoride was added dropwise to the solution of b-D-glucopyranod ^[46] , and then 46 mg (0.26 mmol) of NB
S was added dropwise. The reaction mixture was stirred for 4 hours 30 minutes, quenched with a saturated solution of NaHCO _{3 in} water / ice, extracted with CH ₂ Cl ₂ , washed with brine and dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-EtOAc, 4: 1)
Gave a quantitative yield of 30. Rf (toluene-EtOAC, 10: 1) = 0.
56.

[0165]

Phenyl O- (3-O-benzyl-4,6-O-benzylidene-2-deoxy
-2-phthalimido-β-D-glucopyranosyl)-(1 → 6) -2,4-di-
O-benzyl-3-O- (tert-butyldiphenylsilyl) -1-thio-α
29,800 mg (2.68 mmol) of 475 mg (0.688 mmol) of -D-mannopyranoside (31)
Of zirconocene dichloride, 1.39 g (5.37 mmol) of silver triflate and
Powdered 4 molecular sieve, 14 mL CH_TwoCl_Two30 minutes at room temperature
Stir in the dark. At this time, the reaction mixture was cooled to -40 ° C and 6 mL of CH_Two
Cl_Two437 mg (1.3 mmol) of 30 was added dropwise over 30 minutes. 1
After stirring for 30 minutes, the mixture was washed with NaHCO_ThreeQuenched with a saturated aqueous solution of_TwoCl _Two , Washed with brine, Na_TwoSO_Four, Concentrate and chromatograph
(Diethyl ether / cyclohexane, 1: 2) and 31 (654 mg,
82%). Rf (hexane-EtOAC, 3: 1) = 0.36.

[0167]

O- (3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1 → 6) -2,4-di-O-benzyl -3-O- (tert-Butyldiphenylsilyl) -α-D-mannopyranose (32) 105 mg (0.09 mmol) of acetone 31 in the dark at -15 ° C.
To the solution in 8 mL, 24 mg (0.14 mmol) of NBS was added. After 10 minutes, the reaction mixture was quenched with a saturated aqueous solution of NaHCO ₃ , diluted with EtOAC, extracted, washed with brine and dried. Silica gel column chromatography (
Hexane-EtOAC, 3: 1) gave 31 (91 mg, 94%). Rf (
Hexane-EtOAC, 3: 1) = 0.15.

[0169]

O- (3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1 → 6) -2,4-di-O-benzyl -3-O- (tert-Butyldiphenylsilyl) -α-D-mannopyranosyl trichloroacetimidate (33) 53 mg (0.05 mmol) of 32, 0.25 mL of CH ₂ Cl ₂ at room temperature
To the solution in the mixture, 50 mL (0.50 mmol) of trichloroacetonitrile and 7 mg (0.05 mmol) of flame-dried potassium carbonate were added. After 4 hours, the reaction mixture was diluted with CH ₂ Cl ₂ , filtered through celite and evaporated. By silica gel column chromatography (hexane-EtOAc, 3: 1), 33 (53 mg, 8
8%) as a 13: 1 α / β mixture. Data for 33α: Rf (hexane-EtOAc, 3: 1) = 0.36
.

[0171]

Phenyl 6-O-acetyl-2-O-benzyl-3,4-O-isopropylide
1-thio-β-D-galactopyranoside (36) 193 mg (0.48 mmol) of phenyl 2-O-benzyl-3 at 0 ° C.
, 4-O-isopropylidene-1-thio-β-D-galactopyranoside (35) ^[45] Of acetic anhydride in 0.77 mL of pyridine and DMAP (cat.).
11 mL (1.20 mmol) was added dropwise. Stir at 0 ° C for 5 minutes and at room temperature for 90 minutes
After that, the reaction mixture was evaporated. Silica gel column chromatography
With Sun-EtOAC, 4: 1), 36 of the crude (213 mg, quantitative yield) was obtained.
Obtained. Rf (Hexane-EtOAC, 3: 1) = 0.35.

[0173]

6-O-Acetyl-2-benzyl-3,4-O-isopropylidene-D-galactopyranose (37) 115 mg (0.259 mmol) of 36 of -15 ° C, 5 mL of acetone
NBS 60 mg (0.336 mmol) and 5 mL (0.284 m
mol) of water was added. After stirring for 10 minutes, the reaction mixture was quenched with a saturated aqueous solution of NaHCO ₃ , diluted with EtOAc, extracted, washed with brine and dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-EtOAC,
3: 1) to give 37 (86 mg, 94%). Rf (hexane-EtOAC,
2: 1) = 0.17.

[0175]

6-O-acetyl-2-O-benzyl-3,4-O-isopropylidene-D-galactopyranosyl trichloroacetimidate (38) 81 mg (0.230 mmol) of 37 at room temperature CH ₂ Cl ₂ 1.2 mL
To the solution therein, 230.5 mL (2.30 mmol) of trichloroacetonitrile and 76 mg (0.552 mmol) of flame dried potassium carbonate were added. After 5 hours and 45 minutes, the reaction mixture was diluted with CH ₂ Cl ₂ , filtered through celite and evaporated.
Silica gel column chromatography (hexane-EtOAc, 6: 1) gave 3
8α (29 mg) and 38β (76 mg) were obtained (92% overall yield). Data for 38β: Rf (Hexane-AcOEt, 3: 1) = 0.17
.

[0177]

Data for 38α: Rf (hexane-AcOEt, 3: 1) = 0.37
.

[0179]

Phenyl O- (6-O-acetyl-2-O-benzyl-3,4-O-isopropylidene-α-D-galactopyranosyl)-(1 → 6) -2-O-benzyl- 3
, 4-O-isopropylidene-1-thio-β-D-galactopyranoside (39) 64 mg (0.129 mmol) of 38β, 45 mg (0.112 mmol)
Of a 35% activated powder 4 molecular sieve in 2.1 mL of ethyl ether
Stirred at room temperature for 90 minutes. At this time, 155 mL (0.017 mmol) of a solution of trimethylsilyl triflate in ethyl ether (0.108 M) was added. The reaction mixture was stirred for 45 minutes, quenched with triethylamine, diluted with CH ₂ Cl ₂ , filtered through celite and evaporated on vacuo. Silica gel column chromatography (hexane-EtOAc, 3: 1) gave 39α (62 mg) and 39α.
β (10 mg) was obtained in a total yield of 86%. Data for 39α: Rf (hexane-EtOAc, 2: 1) = 0.42
.

[0181]

Data for 39β: Rf (hexane-AcOEt, 2: 1) = 0.33
.

[0183]

O- (6-O-acetyl-2-O-benzyl-3,4-O-isopropylidene-
α-D-galactopyranosyl)-(1 → 6) -2-O-benzyl-3,4-O-
5. isopropylidene-D-galactopyranose (40) 233 mg (0.316 mmol) of 39α, acetone at -15 ° C.
To a solution in 5 mL was added 73 mg (0.411 mmol) of NBS and 6.3 mL (0 mL).
. (348 mmol) of water was added. After stirring for 5 minutes, the reaction was quenched with a saturated aqueous solution of sodium bicarbonate. The mixture was diluted with EtOAc, extracted and washed with brine. Silica gel column chromatography (hexane-EtOAC, 3:
In 1), 40 (203 mg, quantitative yield) were obtained. Rf (Hexane-EtOAC
2: 1) = 0.12.

[0185]

O- (6-O-acetyl-2-O-benzyl-3,4-O-isopropylidene-
α-D-galactopyranosyl)-(1 → 6) -2-O-benzyl-3,4-O-
Isopropylidene-D-galactopyranosyl trichloroacetimidate (4
1) To a solution of 185 mg (0.287 mmol) of 40 in 1.5 mL of CH ₂ Cl ₂ was added 288 mL (2.870 mmol) of trichloroacetonitrile and 80 mg (0.574 mmol) of flame dried potassium carbonate. The reaction mixture was
Stirred for hours, diluted with CH ₂ Cl ₂ and filtered through celite. The solvent was evaporated under reduced pressure and the crude was purified by silica gel column chromatography (hexane-EtOAc,
4: 1) to give 41α (69 mg) and 41β (111 mg) (80% overall yield) Data for 41α: Rf (hexane-EtOAc, 2: 1) = 0.49
.

[0187]

Data for 41β: Rf (hexane-AcOEt, 2: 1) = 0.27
.

[0189]

O- (3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1 → 6) -O- [2,4-di- O-
Benzyl-3-O- (tert-butyldiphenylsilyl) -α-D-mannopyranosyl]-(1 → 4) -O- [6-O- (2-azido-3,6-di-O-benzyl-2) -Deoxy-α-D-glucopyranosyl)]-2,3: 4,5-di-O
-Cyclohexylidene-1-O-menthoxycarbonyl-1D-myo-inositol (42) 281 mg (0.232 mmol) of 33 and 147 mg (0.165 mmol)
), A mixture of powdered 4 molecular sieves in 3.3 mL of ethyl ether was stirred at room temperature for 45 minutes. At this time, 275 mL (0.030 mmol) of a solution of trimethylsilyl triflate in ethyl ether (0.108 M) was added dropwise. The reaction mixture was stirred for 15 minutes, quenched with triethylamine, diluted with CH ₂ Cl ₂ , filtered through celite, evaporated on vacuo and chromatographed (toluene / E
tOAC, 20: 1) to give 42 (261 mg, 81%), 33 (33
44 mg) and 20 (22 mg, 15%) unreacted. Rf (hexane-EtOAC, 3: 1) = 0.46.

[0191]

O- (3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1 → 6) -O- [2,4-di- O-
Benzyl-3-α-D-mannopyranosyl]-(1 → 4) -O- [6-O- (2
-Azido-3,6-di-O-benzyl-2-deoxy-α-D-glucopyranosyl)]-2,3: 4,5-di-O-cyclohexylidene-1-O-menthoxycarbonyl-1D -Myo-inositol (43) 0.92 of tetrabutylammonium fluoride in THF buffered with acetic acid
6 mL of the M solution was added to 71 mg (0.036 mmol) of 42. The reaction mixture was stirred at 50 ° C. for 10 days, then cooled, quenched with water, diluted with CH ₂ Cl ₂ ,
Extracted and dried over Na ₂ SO ₄ . Silica gel column chromatography (hexane-EtOAC, 3: 1) provided 43 (55 mg, 88% yield). Rf (
Hexane-EtOAC, 3: 1) = 0.23.

[0193]

O- (6-O-acetyl-2-O-benzyl-3,4-O-isopropylidene-
α-D-galactopyranosyl)-(1 → 6) -O- (2-O-benzyl-3,4
-O-isopropylidene-α-D-galactopyranosyl)-(1 → 3) -O- [
O- (3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1 → 6)-]-O- (2,4-di-
O-benzyl-α-D-mannopyranosyl)-(1 → 4) -O- [6-O- (2
-Azido-3,6-di-O-benzyl-2-deoxy-α-D-glucopyranosyl)]-2,3: 4,5-di-O-cyclohexylidene-1-O-menthoxycarbonyl-1D -Myo-inositol (44) 94 mg (0.119 mmol) of 41b, 45 mg (0.026 mmol)
A solution of 43 and activated powder 4 molecular sieve in 0.6 mL of ethyl ether was stirred at room temperature for 90 minutes. At this time, 37 mL (0.004 mmol) of a solution (0.108 M) of trimethylsilyl triflate in ethyl ether was added. The reaction mixture was stirred for 45 minutes, quenched with triethylamine, diluted with CH ₂ Cl ₂ , filtered through celite and evaporated on vacuo. 44α (44 mg) by silica gel column chromatography (2 × cyclohexane-Et ₂ O, 5: 2)
And 44β (7 mg) in a total yield of 83% in 45 (2.3 mg, corresponding to 5% of raw material 43), 46 (35.5 mg, corresponding to 38% of raw material 41β), and 47 (
2.8 mg, corresponding to 8% of the raw material 41β). 44α: Rf (hexane-EtOAC, 3: 1) = 0.26.

[0195]

44β: Rf (hexane / EtOAC, 3: 1) = 0.25.

[0197]

45: Rf: 0.49 (hexane / EtOAC, 2: 1).

[0199]

46: Rf: 0.27 (hexane / EtOAC, 2: 1).

[0201]

47: Rf: 0.38 (hexane / EtOAC, 2: 1).

[0203]

O- (6-O-acetyl-2-O-benzyl-3,4-O-isopropylidene-
α-D-galactopyranosyl)-(1 → 6) -O- (2-O-benzyl-3,4
-O-isopropylidene-α-D-galactopyranosyl)-(1 → 3) -O- [
O- (3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-phthalimido-β-D-glucopyranosyl)-(1 → 6)-]-O- (2,4-di-
O-benzyl-α-D-mannopyranosyl)-(1 → 4) -O- [6-O- (2
-Azido-3,6-di-O-benzyl-2-deoxy-α-D-glucopyranosyl)]-2,3: 4,5-di-O-cyclohexylidene-1-O-acetyl-1
A solution of -D-myo-inositol (48) 44a (11.5 mg, 4.94 mmol) in 3: 2 tetrahydrofuran-methanol (2.2 mL) was added at room temperature to a 2.1 M aqueous lithium hydride (0.
(35 mL, 0.741 mmol). After 31 hours, the reaction mixture was diluted with dichloromethane, washed with water, the aqueous phase was washed with dichloromethane, the combined organic phases were dried over sodium sulfate and co-evaporated twice with toluene. The residue was suspended in chloroform (0.25 mL) and triethylamine (0.7 mL, 4.94).
mmol) was added to the suspension. After cooling at 0 ° C., acetic anhydride (0.12 mL,
1.23 mmol) and dimethylaminopyridine. While keeping the reaction mixture at room temperature for 6 days, triethylamine and acetic anhydride were added every 24 hours. The reaction mixture was diluted with dichloromethane, washed with water, the aqueous phase was washed with dichloromethane, and the combined organic phases were washed with a saturated aqueous solution of sodium chloride, dried over sodium sulfate and evaporated. The residue was purified by chromatography. The NMR spectrum of the reaction mixture showed a 5.5: 1 mixture of 48 and the intermediate compound, so the mixture was dissolved in chloroform (0.25 mL) and triethylamine (0.7 mL, 4.94 mmol) Treated, cooled at 0 ° C., treated with acetic anhydride (0.12 mL, 1.23 mmol) and dimethylaminopyridine,
Warmed to 40 ° C. The reaction mixture was kept at this temperature for 4 days, cooled to room temperature and obtained 8.4 mg (78%) of 48 as above.

[0205]

O- (6-O-acetyl) -2-O-benzyl-3,4-O-isopropylidene-α-D-galactopyranosyl)-(1 → 6) -O- (2-O -Benzyl-3,
4-O-isopropylidene-α-D-galactopyranosyl)-(1 → 3) -O—
[O- (2-acetamido-3-O-benzyl-4,6-O-benzylidene-2]
-Deoxy-β-D-glucopyranosyl)-(1 → 6)-]-O- (2,4-di-O-benzyl-α-D-mannopyranosyl)-(1 → 4) -O- [6-O − (
2-azido-3,6-di-O-benzyl-2-deoxy-α-D-glucopyranosyl)]-2,3: 4,5-di-O-cyclohexylidene-1-O-acetyl-
To a solution of 1-D-myo-inositol (49) 48 (8.4 mg, 3.84 mmol) was added n-butyl alcohol (0.
. 77 mL) and ethylenediamine (167 mL, 2.50 mmol) were added at room temperature. After 90 minutes, the temperature was raised to 90 ° C. and the reaction mixture was kept at this temperature for 18 hours, then cooled, evaporated and the residue co-evaporated twice with toluene. The residue was dissolved in chloroform (0.25 mL) and triethylamine (0.8 mL, 5.7).
6 mmol), cool the solution to 0 ° C., and add acetic anhydride (0.25 mL, 2.6
9 mmol) and a catalytic amount of dimethylaminopyridine. The reaction mixture was warmed and kept at room temperature for 20 hours, then the solvent was evaporated and the residue was purified by column chromatography (2: 1 hexane-ethyl acetate) to give pure 49 (7.5 mg, 9 mg).
3%).

[0207]

O- (2-O-benzyl-3,4-O-isopropylidene-α-D-galactopyranosyl)-(1 → 6) -O- (2-O-benzyl-3,4- O-isopropylidene-α-D-galactopyranosyl)-(1 → 3) -O- [O- (2-acetamido-3-O-benzyl-4,6-O-benzylidene-2-deoxy-β -D-
Glucopyranosyl)-(1 → 6)-]-O- (2,4-di-O-benzyl-α-
D-mannopyranosyl)-(1 → 4) -O- [6-O- (2-azido-3,6-
Di-O-benzyl-2-deoxy-α-D-glucopyranosyl)]-2,3: 4
To a solution of 5,5-di-O-cyclohexylidene-1 D-myo-inositol (50) 49 (9.8 mg, 4.67 mmol) in 3: 7 tetrahydrofuran-methanol (0.5 mL) was added methanol. Sodium methoxide.
A 2M solution (70 mL, 14.0 mmol) was added at room temperature. After 8 hours, the reaction mixture was neutralized with Dowex 50WX resin, filtered and evaporated. The residue was purified by column chromatography (3: 2 hexane-ethyl acetate) to give 50 (8.
(6 mg, 91%).

[0209]

[0210]

[References]

[Brief description of the drawings]

1 shows a chemical formula of a preferred hexasaccharide according to the first aspect of the present invention, and 2 shows a chemical formula of a GPI anchor typically present on a cell membrane.

FIG. 2 shows retrosynthesis analysis from the synthesis methods of the eighth to tenth aspects of the present invention.

FIG. 3 shows a scheme of a synthesis method of the present invention.

FIG. 4 shows a scheme of a synthesis method of the present invention.

FIG. 5 shows a scheme of a synthesis method of the present invention.

FIG. 6 shows a scheme of a synthesis method of the present invention.

FIG. 7 shows a scheme of a synthesis method of the present invention.

FIG. 8 shows a scheme of the synthesis method of the present invention.

FIG. 9 shows a scheme of a synthesis method of the present invention.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 31/7048 A61K 31/7048 45/08 45/08 A61P 3/10 A61P 3/10 21/00 21 / 00 25/00 25/00 25/08 25/08 25/14 25/14 25/16 25/16 C07H 9/04 C07H 9/04 11/00 11/00 13/06 13/06 15/04 15 / 04 F 15/203 15/203 17/04 17/04 (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE) , LS, MW, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CR, CU, CZ, DE, DK, DM, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT , LU, LV, MA, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (72) Inventor Jose Luis Tiara Spain E-2806 Madrid 3, Juan de la Cielva F-term (reference) ) 4C057 BB07 CC01 DD01 FF03 GG10 HH03 JJ10 JJ23 KK02 4C084 AA17 NA14 NA15 ZA01 2 ZA032 ZA062 ZA152 ZA162 ZA942 ZC352 4C086 AA01 AA03 EA01 EA03 EA10 MA01 MA04 MA09 NA14 NA15 ZA01 ZA03 ZA06 ZA15 ZA16 ZA94 ZC35

Claims (28)

[Claims], 1. A compound of the general formula I: Wherein each R is independently selected from hydrogen, an alkyl or substituted alkyl group, an acyl or substituted acyl group, a phosphate group or a protecting group, or two R groups are cyclic A NX group represents N ₃ ; or in the NX group, X is independently selected from hydrogen, alkyl or substituted alkyl, acyl or substituted acyl
One represents the above groups; -NY represents a phthalimido group (NPHt) or N _3; or in NY, Y is selected from hydrogen, alkyl or substituted alkyl, independently acyl or substituted acyl Wherein at least one group is represented by the following formula: wherein the number of phosphate groups in the molecule is 3 or less, or a salt or derivative of the compound. 2. The compound according to claim ^1, wherein R ^1a and / or R ^6f is a phosphate group. 3. The compound according to claim ^1, wherein R ^1a and R ^2a together are a cyclic phosphate group. 4. The compound according to claim 1, wherein NX is an NH ₃ ⁺ group. 5. The compound according to claim 1, wherein Y is an AcH group. 6. The compound according to claim 1, wherein the R group in one or more of the units a, b, c and / or d is hydrogen or a phosphate group. 7. The compound according to claim 1, wherein the derivative is a coordination complex of the compound and a metal ion. 8. The compound according to claim 1, wherein the derivative is a prodrug. 9. The compound according to claim 8, wherein said prodrug is a glycolipid derivative which can be converted to I after phospholipase cleavage, and R ^1a is O ‖ -PO-diacylglycerol ‖O. 10. The compound according to claim 1, wherein said compound is bound to a binding partner. 11. The compound according to claim 10, wherein the binding partner is a label, a support substrate, a carrier, an effector or inhibitor molecule, or an immobilizing agent. 12. A composition comprising one or more compounds according to any one of claims 1 to 11. 13. Combining one or more compounds according to any one of claims 1 to 11 with a pharmaceutically acceptable adjuvant and / or one or more other therapeutically active agents. A method for preparing a pharmaceutical composition. 13. A compound according to any one of claims 1 to 11 for use in a method of medical treatment. 14. A compound according to any one of claims 1 to 11 for the preparation of a medicament for treating diabetes. 15. The method of claim 15, wherein the diabetes is obesity type II diabetes (NIDDM), diabetes due to insulin resistance, insulin resistance of type I diabetes, and insulin resistance or low insulin such as unstable diabetes and polycystic ovary disease. 15. Use according to claim 14, which is a productivity-related condition. 16. Use according to any one of the preceding claims for the preparation of a medicament for the treatment of nervous system, motor neuron diseases, neurodegenerative disorders or neuropathies. 17. The method according to claim 17, wherein the nervous system damage includes one or more traumas, strokes, surgery,
17. Use according to claim 16, being the result of an infection, ischemia, metabolic disease, or toxic agent. 18. The use according to claim 16, wherein the motor neuron disease is spinal muscular atrophy, paralysis or amyotrophic lateral sclerosis. 19. The use according to claim 16, wherein said neurodegenerative disorder is Parkinson's disease, Alzheimer's disease, epilepsy, multiple sclerosis, Huntington's chorea or Meniere's disease. 20. A compound of the general formula II, III, IV or XI Embedded image Embedded image Embedded image Wherein R, X, and Y are as defined in claim 1 and each L is an elimination that activates the anomeric position of the corresponding sugar unit in the preparation of a glycosylation reaction at a glycosyl receptor. Or a salt or other derivative of the compound. 21. R ^4b (preferably hydrogen, or a protecting group selected to allow orthogonal deprotection to other protecting groups present) is different from R ^3b , preferably different from R ^6b , ^21. The compound according to claim 20, represented by the general formula II, wherein R ^3b and R ^6b are the same as each other but different from R ^4b , or a salt or other derivative of said compound. 22. A method of synthesizing a compound represented by Formula I, comprising: (a) preparing at least two intermediate disaccharide compounds II, III, and IV; and (b) at least two intermediate disaccharide compounds. Condensing II, III, and IV together to form a tetrasaccharide intermediate compound; and (c) combining the tetrasaccharide intermediate with a third disaccharide intermediate selected from compounds II, III, and IV Reacting to produce a compound of formula I. 23. The method of claim 22, wherein the three disaccharide compounds II, III, and IV react together to form a compound of Formula I. 24. The method of claim 22 or claim 23, further comprising removing one or more R protecting groups after formation of the hexasaccharide. 25. The method of claim 22, comprising reacting disaccharides II and II to form a tetrasaccharide intermediate of formula XI, and further reacting the tetrasaccharide intermediate with a compound of formula IV.
The described method. 26. A 2-azido-2-deoxy-D of the formula VI wherein X = N _2.
A glycosyl donor in the form of a glucopyranosyl is converted to 2-amino-2-deoxy D
Preparing from a glucosamine salt by a diazotransfer reaction from methanesulphonyl azide trifluoride; then reacting the donor with a glucosyl acceptor in the form of a building block of myo-inositol of the formula V A method for synthesizing the compound of 27. A method for synthesizing a tetrasaccharide of the general formula XI by combining a glycosyl acceptor of the formula II wherein R ^4b is hydrogen and a glycosyl donor of the formula III wherein L is a leaving group. 28. Compounds 9, 10, 11, 12, 13, 18, 19, 20,
24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 35, 36,
37, 38, 39, 40, 41, 42, 43, 44, 45, 48, 49, or 5
A compound selected from 0.