JPH0471919B2 - - Google Patents

Description

[Detailed description of the invention] The present invention provides novel derivatives and compounds having a uronic acid structure.
and a method for synthesizing the derivative. The invention further relates to the uses of these derivatives, in particular
Intermediates of lycoside synthesis, oxygen substrates, haptens and
Concerning use as an experimental reagent. The present invention particularly relates to D-glucuronic acid and L-ise.
It concerns a derivative of an epimer of rononic acid. Each of these epimers has the formula It is known that The uron nucleus with the above structure has a very important
Participating in the organization of the glycoside chains of biologically active compounds
Ru. The above-mentioned cores may in particular contain various natural mucopolysaccharides,
For example, heparin, heparan sulfate (heparane
sulphate), chondroitin, etc. The applicant has proposed that fragments of mucopolysaccharides as described above or
Oligosaccharides corresponding to fragment derivatives can be synthesized
I've done a lot of research and devised a method. From the results of these studies, it has been found that
For example, glycosylation reaction
provide sufficient reactivity in a given position to participate in
and/or selective substituents at a given position.
The use of certain protecting groups may be necessary to allow the introduction of
It turned out to be essential. Especially when considering the L-iduronic acid structure.
However, only a few examples of derivatives have been described in the literature.
not present. Especially as such literature
Tetrahedron, volume 32, pages 1399–1402, 1976.
Included KISS and WYSS papers and Carb.Res.60
(1978), 315-326, SRIVASTAVA et al.
The following papers are listed. Furthermore, selective
Using oxidation methods results in low yields and poor reproducibility of results.
do not have. Therefore, it is difficult to link L-iduronic acid nuclei to expensive compounds.
The methods described above cannot be used to
will be understood. Currently, the active ingredient uronic acid derivatives are
Glycosylate reagent in cosylation reaction
It is inconceivable that it would be possible to react as
stomach. The inventors have therefore determined that suitable nuclei, particularly suitable for said components,
D-glucuronic acid and L-iz
The structure of ronic acid itself and the desired derivatives are easily obtained.
It was necessary to research a method that would allow this. Therefore, an object of the present invention is to improve the glycosylation reaction.
glycosylate reagent or glycosylated
reagents (in common parlance, so-called aglycones)
sufficient responsiveness to be able to participate as a
Novel D-glucuronic acid and L-iduronic acid with
An object of the present invention is to provide derivatives of phosphoric acid. Furthermore, the object of the present invention is to
to specifically introduce selected substituents into predetermined positions.
The object of the present invention is to provide novel derivatives that have the following properties. Furthermore, the object of the present invention is to improve the glycosylation reaction.
glycosylate reagent or glycosylated
It has sufficient reactivity to be used as a reagent.
and selected at a given position depending on the nature of each protecting group.
New derivatives that allow selective substituents to be specifically introduced
It is to provide. Furthermore, one object of the present invention is to determine the structure of the acid in question.
In particular, forming the structure of L-iduronic acid,
Provides a method for easily producing reactive derivatives
That's true. The object of the invention is furthermore to provide glycoside components, especially biological
Reagents especially useful as oxygen substrates and/or experimental reagents
These derivatives can be used to develop new glycosides.
It is to use it. A further object of the present invention is to bind to high molecular weight substances.
Manufacturing antigenic determinants that induce antibody formation in animals
By using these derivatives to
Ru. The object of the present invention is further directed to said antibody or said antibody.
purify any other substance that has an affinity for
the above to produce immunoabsorbents particularly useful for
is to use a conjugate of The derivative of the present invention is formula(); [In the formula, the substituent -OR₁～−CR_Fiveis a reactive group, i.e.
Indicates an active group or a functionalizable group, and the reactive group or functionalizable group is
The active group is the glycosylate reagent glycosylate.
Perform the glycosylation reaction as a reagent
For example, halides, especially bromides or
or chloride, -O-imidoyl group (-O-
imidoyl group);
The root ester group is or a hydrogen atom, and the
The enabling functional group constitutes the beginning of the conjugable arm.
For example, an -O-alkenyl group such as -O-a
lyl or -O-vinyl, or protected form
The conjugatable arm itself is located in the −OH functional group.
The groups are blocked by protecting groups and are identical or
are different from each other and are different from each other and from one or more reactive groups.
Monosatsukara that can coexist and is performed when necessary
is inert with respect to reactions involving the
Protecting groups are preferably aliphatic or aromatic radicals.
selected from Cal, more preferably having 1 carbon atom
-4 alkyl radicals, especially methyl groups, substituted alkyl radicals
Kill radicals such as benzyl radical, acyl radical
Radicals such as acetyl, benzoyl and chloroa
Cetyl and alkenyl la having 1 to 4 carbon atoms
groups consisting of radicals such as allyl and vinyl
selected from. (However, methyl (benzi)
L-2,3-di-O-benzyl α,β-L-ide
With the exception of viranoside) uronate,
In the D-glucuronide compound, R₂,R₃and R_Fourteeth
All three are not the same group, and R₁and R_Fourteeth
Methyl group (this group is difficult to remove)
uronic acid derivatives or their
It is salt. The derivative of the present invention having an iduronic acid structure is semi-
- Including open derivatives and open derivatives
I want to be noticed. The term semi-open derivative refers to
induction activated or potentially activated by
means the body. The semi-open derivative on the right is in the 1st position during the reaction.
It is a derivative to which another compound can be attached. Left semi-open derivatives can participate in the reaction
Containing only one free functional group, in particular only one -OH group
nothing. Open derivatives are semi-open on both the left and right sides.
Yes, so optionally glycosylate reagents or
Can be used as a glycosylated reagent. In this case, the above-mentioned derivatives may be temporarily (temporarily)
protective group, i.e. regardless of other protective groups present.
Protecting groups that can be removed by alcohol reformation
Contains. The derivative of the present invention having a glucuronic acid structure is
It is open. According to another advantageous feature of the invention, the protecting group R₂
and R₃and optionally R_Fourare different from each other.
without changing the acid structure and carboxyl functionality.
The next thing is that it can be removed. The advantage of this feature is
Different from each other in place of all protecting groups in successive reactions
Possibility of introducing substituents or one or more substituents
Introducing the substituent and releasing the -OH group at the remaining position.
It's about getting. In preferred derivatives of the invention, the reactive group is 1
It exists in the position. In another preferred derivative, the reactive group is in the 4-position.
exist. A preferred derivative of the present invention is D-glucuronic acid.
or its salt, and the glucuronic acid is
formula(); [In the formula, -OR₁is a reactive group, and (a) glycosyl
The glucuronic acid derivative is used as a glycosylation reagent.
Active groups that can participate in sylation reactions, e.g.
halides, especially chlorides or bromides,
-O-imiddle group or R₂Ortho-S combined with
(b) functional or functionalizable groups such as -O-
Alkyl radicals, especially O-methyl or -O-
benzyl, -O-alkenyl e.g. -O-allyl
or -O-vinyl, or -O-acyl, especially -
selected from O-acetyl or (c)-OH group, R₂
~R_Fourare the same or different, and include protecting groups such as carbon atoms.
Alkyl radicals with 1 to 3 molecules, especially methyl, substituted
Alkyl e.g. benzyl, acyl e.g. acetyl
benzyl or chloroacetyl, or alkaline
Kenyl represents for example allyl or vinyl, R_Five
is an aliphatic or aromatic radical, especially one containing 1 or more carbon atoms.
4 alkyl radical or substituted alkyl e.g.
] (however, D-glucuronic acid excluded above)
except for). Among uronic acid derivatives R_Fouris tentatively called −OH group.
to form an ester or ether to retain the -OH group.
It is a temporary group that protects monochloroacetyl, acetyl
chill, benzyl or p-methoxybenzyl
Radical, or allyl or vinyl type radical
Preferred is the group selected from Cull et al. Preferred products of this group contain R₁
represents an allyl group or a vinyl group. Generation as above
The advantage of this product is that the substituents contained in the 1st and 4th positions are
Selective without changing the groups at 2, 3 and 6 positions
This means that it can be removed. In another preferred group R₁is a hydrogen field
and the hydrogen atom can be substituted with a halogen atom if desired.
Introducing a reactive group such as ide or imitate into the 1-position
or glycosylation reaction.
It can occur. In another preferred group, R₁But for example
For example, monosaccharides can participate in glycosylation reactions.
represents an active group as described above. Another preferred group of products exhibits hydrogen atoms.
SuR_Fourand in this case R₁is a government official like the one mentioned above.
It is advantageous to indicate an activatable group. In these various groups R₂and R₃What is
They may also be equivalent groups, for example aryl groups, especially
Indicates a benzyl group. or R₂and R₃may be mutually different groups. Another derivative of the invention has the formula (); [In the formula, the substituents are as defined for formula ()
The L-iduronic acid inducer represented by
It is a conductor. In a preferred group of L-iduronic acid derivatives
R₁and R₂indicates orthoester. R in another preferred group₁is halide
Indicates an active group such as In yet another preferred group R₁Haa
Indicates alkyl group or alkenyl group. Preferred products of these groups have a temporary position in the 4th position.
Advantageously, constant protecting groups are included. Another particularly preferred product of these groups is
Furthermore, mutually different substituents R₂and R₃including
These substituents have various effects during continuous processing.
This allows for various placements on the constituent units.
Substituents may be introduced. For example, R₂indicates an acyl group such as an acetyl group.
ShiR₃When represents a benzyl group, an acetic acid is added to the 2-position.
A sulfate group is introduced in place of the chill group, and then benzene
-OH at position 3 blocked by group
It is possible to liberate the group. Therefore, the structure of heparin or heparan-sulfate chains is
component units are advantageously obtained. The substituents of the present invention may further be used in the synthesis of these derivatives.
The purpose is to provide a method. According to the synthesis method of the present invention, they can coexist with each other.
Monosaccharide with a uronic acid structure as a protecting group
into a reactive group or, if necessary, a given protecting group.
It is possible to selectively process the positions where you want to introduce
Make it. When used during the synthetic process, the group is an intermediate protecting group.
This group is called a reactive group or a final protecting group, i.e.
A group corresponding to a desired protecting group is added to the monosaccharide of the present invention.
removed to place in place. Introducing one or more final protecting groups from the beginning
It is possible to enter. Also, for the introduction of one or more intermediate protecting groups.
Therefore, it is first possible to process a given position, and this
Once the desired group has been introduced at the position, one or more of the above
one or more intermediate groups occupied by
is intended for the introduction of final protecting groups at multiple positions with different positions.
In some cases, it is processed continuously. As an example, producing monosaccharides containing vinyl groups
, all the -OH groups of its alcohol functional groups are
by the final protecting group or in some cases intermediate
Allyl derivatives blocked by protecting groups
It is advantageous to form a This allyl monosaccharide
When an alkylating agent is applied to
The -OH group of the functional group is blocked. Next is this
An example of a monosaccharide with an allyl group protected by
For example, in the presence of diazoabicyclooctane
The corresponding vinyl ether
becomes le. If a temporary group is introduced at a given position, then this
Removes the temporary group of glycosylation reaction.
The useful alcohol is re-formed at this location, while the alcohol
The Nyl group undergoes one or more reactions.
Note that the chains can be distracted. From protected 1-O-vinyl derivatives, e.g.
Reactive groups such as baimidate can be formed. this
To do this, add mercuric oxide and chloride to vinyl derivatives.
It is possible to apply the action of mercury derivatives such as mercuric mercury.
This action allows the alcohol to be reformed.
This alcohol is trichloroacetonyl
It is affected by imidoylating agents such as tolyl. Also, the acids of glycosides, especially methyl glycosides
1-O-hydroxy obtained by hydrolysis
It is also possible to obtain imidates from fluorinated derivatives.
be. Monosaccharides usually have a halogenated uronic acid structure
is advantageous due to the action of the halogenating agent on the acyl group.
The acyl group itself is obtained, for example, in alcohols.
Obtained by acylation. virusmail
(Vilsmeir) reagent to 1-hydroxy derivatives.
It is also possible to obtain the above-mentioned monosaccharides by using Furthermore, monosaccharides containing orthoester groups can be
prepared by an advantageous operation from halogenated derivatives of
It is also possible to build Depending on the preferred manufacturing method
For example, halogenated derivatives such as sym-collidine
Alcohol acts in the presence of proton receptors
Ru. According to an advantageous characteristic of the invention, the L-Izuron construction
The monosaccharide itself is D-glucofura
form from the noside type derivative by the following treatment.
Methyl 3-O-benzyl-L-iduronate formed
Manufactured from formula(); [In the formula, R₆~R₈The groups may be the same or different groups.
[alpha]-D-glucofuranoside showing a locking group]
is used. Especially R₆and R₇forms an isopropylidene group
and R₈is a benzyl group. By treating the monosaccharide of the formula, -OH at the 5th or 6th position
selectively act on one or the other of the groups;
The 5th position was changed from D-gluco structure to L-ide structure.
by converting the 6th position from the primary alcohol structure to the carboxylic acid.
Changes to phosphoric acid structure. The rearrangement of furanose form to pyranose form is as follows.
It is a cage. -OH group of α-gluco chain is selectively treated to form
Id from the glucofuranoside ring using the constructed chain.
Changed to furanoside ring, then idopyranoside ring
is formed as follows. -CH in 6th and 5th positions₃OH group and -CHOH group
Continuously change each as follows: 1-CH₂OH to −CH₂Transition to OTr. here
Tr represents a trityl group. The starting material α-glucofuranoside was treated with
to selectively block the -OH group at position 6.
For this purpose, treatment with trityl chloride etc.
That is advantageous. 2 -CHOH to -CHOR₉Transition to. Here R₉teeth
It is a blocking group. A preferred blocking group is R₃is a benzoyl group
This corresponds to the case where . Introduction of benzoyl group
For this, the benzoyl chloride group is used. 3-CH₂OTr to −CH₂Transition to OH. Deblocking of hydroxyl groups is
With luenesulfonic acid, or alternatively,
Performed using a methanol solution of boron fluoride, etc.
It is advantageous to do so. 4-CH₂Transition from OH to −COOH. For this step, the carboxylic acid form
using reagents that allow the migration of This conversion uses a sulfuric acid solution of chromic oxide.
Advantageously, this is carried out by using 5 - Transition from CHOBz to rCHOH. Treat the product obtained in the previous step with soda
Ru. This is further converted into the acid form. Cation exchange resin to remove cations
It is advantageous to use 6 -Transition from -COOH to -COOMe. here
Me represents a methyl group. Usually, especially by the action of diazomethane,
Perform tell. 7 Transition from -CHOH to -CHTf. Here Tf
represents a trifuryl group. 5th place, tosylate, mesylate or preferred
or highly reactive compounds such as trifuryl groups.
Introduce a group. Then, anhydrous trifuryl
Perform processing. 8 Transition from -CHOTf to -CHOTFA. TFA
is trifluoroacetate group (carbon at position 5 is inverted)
(in the form of Nucleophilic substitution is performed by Walden inversion.
This reaction was performed using sodium trifluoroacetate.
It is advantageous to implement. 9 Transition from -CHOTFA to -CHOH. The product was treated with methanol followed by purification.
Therefore, it is advantageous to apply chromatography.
Ru. 10 Ring dislocation. Using an acid medium, especially in the presence of trifluoroacetic acid
That is advantageous. A mixture of trifluoroacetic acid/water (approximately 9/1) is especially
Are suitable. The present invention also uses D-gluco and
and furanosides having an L-ide structure and the above-mentioned
Idopyranoside obtained at the end of steps 1 to 8
The purpose is also In particular, R_Fourrepresents an alkyl group, especially methyl,
Regarding the compound with the L-iduronic acid structure of the obtained formula
This compound is then subjected to saponification and catalytic hydrogenation reactions.
Note that L-iduronic acid units are produced when
I want to be. The above compounds may also be acylated and halogenated
It may be converted into a compound or even an orthoester.
stomach. The protecting group at position 4 can be removed, thus leaving a separate
Temporary protecting groups can be introduced. By appropriately selecting the protecting group at the 4-position,
Depending on the nature of the orthoester, this
The compound is subjected to glycosylation reaction, and then
The protecting group at position 4 can be selectively removed. this
Another glycosylation reaction can take place at the
Ru. Finally, the ester or acyl formed at the 2nd position
The group is removed during the first glycosylation reaction
and therefore, if desired, can be sulfate -
The OH functionality can be regenerated. According to another embodiment, the monosaccharide of L-idose is
Prepared with desired protecting groups at positions 1, 2, 3 and 4
, the protecting group is left unaffected here, and the 6-position
The primary alcohol functionality of is selective towards carboxylic acids.
oxidized to Monosaccharides with a D-glucuron structure are particularly
by using conventional methods of introducing the desired group.
It is produced from a derivative of glucose. Regarding the methods used so far, the above
defined derivatives, especially those with L-iduron structure
The means used in the present invention to obtain the derivatives are:
It is said to be easy to implement and produce reproducible results.
has advantages. Due to the great flexibility of these methods,
Based on the reactivity of the chain, it has a favorable stereochemistry.
Uro that can give rise to various types of chains
It is possible to obtain derivatives having the following structure.
Furthermore, depending on the type of protecting group chosen, a given
After going through a type of reaction, the location of the protecting group
selectively introducing a given substituent into and/or
can liberate one or several -OH groups.
It is Noh. In this way, e.g. the biologically active
Specially substituted molecules such as those encountered in molecular chains
Obtaining oligosaccharides containing uron units
is possible. In this regard, 2-O-sulfate-L-iduro
The method of use for deriving phosphoric acid derivatives is particularly
be recognized. In addition, the present invention provides oxygen substrates for glycosidases, etc.
The present invention is useful for the synthesis of biologically important glycosides.
The purpose is to use uronic acid derivatives. Such glycosides are suitable for example as derivatives of the invention.
paranitrophenol or methylberifue
by condensation with ron or their derivatives.
can be obtained. For this purpose, the Koenitz
Koenigs-Knorr reaction, etc.
reaction can be used. In this case, the formed
In the presence of a reagent that fixes the halohydric acid
It is desirable to combine oside with
aglycon and Oside Harai
contact with oside halide. Regarding iduronic acid, the uronic acid structure has already been formed.
glycosidic bonds are formed at the same time as
is particularly noteworthy. This feature depends on the product, or protein or solid support.
by facilitating the glycosylation of substances such as
Ru. These products or substances have the iduronic acid structure
As mentioned above, the glycosidic bond is formed before forming
It is not possible to maintain the synthesis step of the prior art method.
stomach. Enzymes derived from monosaccharides of the present invention
The substrate is conveniently a variety of enzymes, especially Heparin.
The specificity of the enzyme that decomposes L-iduronine, especially
This allows examination of the specificity of the dasase. For example, the present invention on methyllambelliferon
Glycoside obtained by fixing monosaccharide
constitutes a complex compound that is an effective substrate for fluorescence.
Ru. In another application, the monosaccharide of the present invention
Used in the development of radioactive substrates. These substrates are
By using radioactive starting glycosides,
is the above-mentioned nitro-phenyl-glyco
obtained by reducing the side group to an amine group.
and then e.g. tritium using conventional techniques.
can be subjected to an acetylation reaction with acetic anhydride.
Ru. Obtaining derivatives that already have a uronic acid structure
The importance is once again being noticed. In fact, radioactive compounds
Considering that it is expensive, a given glycoside
radioactive compounds must be processed to form
The goal is to reduce the number of steps that are not required as much as possible.
You can see how important it is. According to another aspect of the invention of critical importance, anno
functionalisable groups on the mercantile carbon.
Monosaccharides, including group), are found in various organisms.
Fixed bond (anchoring arm) useful for scientific applications
This provides an avenue for introducing chains that can play the role of According to yet another embodiment, the glycoside is capable of
In some cases, it is converted into a fixed bond, and these bonds are
Facilitates immobilization to proteins through binding, and
This facilitates the acquisition of artificial antigens. For this, the above-mentioned para-nitro-glycosides
The reduction reaction of glycosides such as uron derivatives
In this way, the corresponding para-amino-phthalate
leading to enyl glycosides. Next, this glycoside
can be used to couple proteins through a conventional diazo coupling reaction.
Aromatic moieties in white matter, especially bonds involving tyrosine moieties
be involved in Can be purified and used as an immunological reagent
In order to induce antibody production that can be used
Such antigens are used in animals. In this way, glycosides as mentioned above
or functionalizable substituents of uronic acid derivatives.
et al., especially (1) Protected or functional groups, if necessary.
2 to 10 containing -OH group bonded to at the chain end
an alkyl group consisting of carbon atoms, (2) Contains alkenyl groups containing 2 to 10 carbon atoms
It is possible to form various fixed bonds.
Ru. Type (1) chain is alkylene-glycol
group, α,β-dihydroxy-propyl group, or β
-Hydroxyethyl group, γ-hydroxypropyl
Conveniently applicable to the group of
It is for use. Chains (1) and (2) above can be further modified by one or more
ter and/or intermediate amine and/or amine
Nitrogen as a functional group, terminal ether group, terminal carboxyl group, etc.
Functional powder containing xyl group or terminal aldehyde group, etc.
Conveniently it includes a terminal group. Particularly preferred amino-terminated chains are
It is of the xy-alkylamine type. this place
In general, the alkoxy group and the alkyl group are 3-
5 carbon atoms, preferably 4 to 10, suitably
Contains about 10 carbon atoms. Agence Nationale de Valorisation de la
J.A. Patent Application No. 500626/79 under the name Recherche
According to the method disclosed in
Advantageously obtained from the allyl derivatives of the invention. The reactivity of this group actually makes it suitable for a number of reactions.
Adjustable length and gender
hydrophilic properties important for biological applications.
allows the acquisition of chains with To lengthen the chain, traditional reactions in organic synthesis are used.
I have to rely on it accordingly. These reactions can be performed by those skilled in the art.
It can be easily used to create the desired chain by
used. For example, the alcohol function
chain by ozonolysis of the double bond of the group
, then toshile, and finally this toshile.
It is possible to react the derivatives with azide.
Ru. The azide obtained is of double importance.
This azide is a stable compound on the one hand and
Then, the immune system of the present invention can be prepared by reduction in the usual manner.
Particularly important in the composition of immunoadsorbants
Facilitates the production of substituted chains with terminal amino groups.
Ru. Anchoring arm as described above
Monosaccharide with arm) is particularly effective for immunoabsorption.
A form fixed on a solid carrier to constitute an adhesive
constitutes an effective and valuable biological reagent. These immunoadsorbents, such as those obtained above,
body, or any agent that has affinity for these antibodies.
It can be used in particular for the purification of substances of interest. These immunoadsorbents take advantage of this affinity
It can also be used in the course of medication. Suitable solid carriers include, for example, cellulose, agar, etc.
rose or cross-linked agarose (especially Pharmacia
The product name Cephalose is commercially available from
), or in addition polyacrylamide (especially
Ultrogel and Magnogel trademarks by IBF
The above-mentioned polysaccharides such as those sold under the trademark
By mixed polymers containing ride polymers
configured. Furthermore, the monosaccharide of the present invention is polyclonal.
For the purpose of producing sexual or monoclonal antibodies
BSA (bovine serum albumin) or polylysine, etc.
Enables the construction of haptens that can be attached to soluble carriers of
Make it. Besides uses such as antigen determination, the products of the invention
is also important as a comparison product for structural studies.
Ru. Advantages of other features of the invention can be seen in the examples below.
By referring to the attached drawings of Figures 1 to 16,
It becomes clear. Also, the references used in these figures to indicate derivatives are
Reference numerals are used again in the examples. The structural formulas of the compounds of Examples 1 to 6 are shown in Figure 1 and
Shown in Figure 2. Example 1 (Allyl-4-O-acetyl-2,3-di-O
-benzyl-α-D-glucopyranoside) uro
Production of acid (compound 8a) According to the following steps a) to g), glucose
Perform this synthesis. a Allyl α-D-glucopyranoside (compound
1) Manufacturing Gaseous hydrochloric acid (18g) to allyl alcohol
Heat the solution (600 ml) to 70 °C. next
Add anhydrous glucose (300g) and incubate for 3 hours.
Maintain temperature. After reaction methanol/chloroform (1/4,
v/v) thin layer chromatography in solvent (t.l.c.)
get. The brown solution obtained after 3 hours was placed in vacuo.
Dry, concentrate and reconstitute with concentrated ammonia solution (50ml).
Mix and dry and concentrate again. to the resulting residue
Add acetone (500ml) and bring to a boil until the entire amount is dissolved.
Maintain this state until it is resolved. After cooling, this liquid
Decant the body. Repeat the same treatment on the residue.
cormorant. Residues of derivative 1 by T.L.C. analysis of extracts
or become very strong due to impurities.
Continue the above treatment until contaminated extracts are identified.
cormorant. A portion (12 g) of the extracted first fraction was siluted.
Apply color chromatography. Acetone/Ethe
Derivative 1, which can crystallize in the mixture, is recovered.
(6.5g; M.P. 95-99°C). The remainder of the product was passed through the same
It can be purified in steps. b) Allyl 4,6-O-benzylidene-α-D
-Production of glucopyranoside (compound 2) Compound 1 (37g) was dissolved in dimethylformamide (200g).
ml). Then dimethoxytoluene (41
g) followed by hydrated paratoluene sulfonate.
Add phosphoric acid (130mg). After heating under vacuum and reflux for 2 hours (in a water bath), the reaction
(t.l.c. methanol/chloroform,
2/23, v/v). Evaporate the solvent. syrup
Dissolve in methanol (minimal amount). This solution
in an aqueous solution of sodium bicarbonate (6.3 g in 320 ml of water)
drip onto. The obtained precipitate was dissolved in ethanol.
Recrystallize (21 g; M.P. 120-121°C). The mother liquor is
Further product 2 is given. Overall yield (37g; 71.4%). c) Allyl 2,3-di-O-benzyl-4,6
-O-benzylidene-α-D-glucopyranosyl
Production of compound 3 Compound 2 (45g) was dissolved in anhydrous DMF (500ml)
I let it happen. Sodium hydride (50% dispersion in oil 28
Add g). After 30 minutes, the mixture was cooled to 0°C and then treated with benzene bromide.
Zil (52ml) was added dropwise. After reaction T.L.C.
(ether/hexane, 1/1, v/v)
Ru. Next, slowly add methanol (150ml),
Evaporate to dryness and remove again with chloroform.
vinegar. Wash the chloroform phase with water and dilute with sodium sulfate.
Dry on a dry surface. After evaporation of the solvent, the residue is
crystallize in a tel/hexane mixture (36.5
g; M.P. 83-84°C). The product is t.l.c. (ether/hexane, 1/
1, v/v) to impurities that migrate higher in
Therefore, it is slightly contaminated. d) Allyl 2,3-di-O-benzyl-α-D
-Production of glucopyranoside (compound 4) Compound 3 (56g) in methanol (1) solution
Add water (450ml) and then add the hydrated paratolue.
Add sulfonic acid (17g). After 2 hours at 80°C, the mixture was allowed to cool and the solution
Evaporate the liquid and dissolve the residue in chloroform (1
) to take it out again. Chloroform solution has medium pH
Rinse with water until dry, then wash with sodium sulfate.
dry. In this way, pale yellow syrup (48g) was obtained.
Ru. This syrup is used in the next step (synthesis of compound 5).
used for. e Allyl 2,3-di-O-benzyl-6-O-
Trityl-α-D-glucopyranoside (compound
5) and its 4-O-acetylated analogs (compound
Item 6a) The obtained derivative 4 (48 g) was added to pyridine (250 ml).
and add trityl chloride (38.5g).
Ru. After 1 hour at 100°C, the reaction is terminated (t.l.c.
ether/hexane, 1/1, v/v). melting the tip
Add acetic anhydride (200ml) to the solution. The next morning, the reaction
complete it. (t.l.c., ether/hexane, 1/
2, v/v). Evaporate to dryness and chloropholate the residue.
Remove again with lume (500 ml) and remove chloroform phase.
with 10% acidic potassium sulfate solution and then with water.
Then dry with sodium sulfate. Evaporate the chloroform. In this way
Compound 6a is obtained. Produce compound 7a as is.
Used in reaction. f) Allyl 4-O-acetyl-2,3-di-O
-benzyl-α-D-glucopyranoside (compound
Manufacture of item 7a) The obtained derivative 6a was added to chloroform (500ml).
Dissolve. Cool this solution to 0°C and
Add a methanol solution of boron trifluoride (20%, 120
ml) dropwise with stirring. After this reaction
t.l.c. (toluene/acetone, 10/2, v/v)
Let's do it. Pour the reaction mixture into a small flask (separatory funnel) for decanting.
Transfer to Wash the chloroform phase with water
(2×100 ml) and a saturated solution of sodium bicarbonate.
and then with water until the PH is neutral.
Ru. After drying and evaporation, the resulting residue was dissolved in toluene.
onto a column of silica gel (500 g) equilibrated in
Introduce. Most of the impurities are removed by pure toluene
After eluting the product in a toluene/acetone mixture
(10/2, v/v). like this
to obtain 48 g of compound 7a. Combine this directly
Used to synthesize Object 8a. A portion of compound 8a is obtained in pure form.
[α]²⁰ _D= +11° (chloroform). This I.R. and N.
According to M.R. spectrum, elemental analysis shows the same
Check the structure. g) (allyl-4-O-acetyl-2,3-di
-O-benzyl-α-D-glucopyranoside)
Production of uronic acid (compound 8a) Acetone (800ml) solution of compound 7a (48g)
Cool to -5°C. Next, add chromium trioxide (30g)
A solution of sulfuric acid (35M; 125ml) is added dropwise. Mixed
Leave the mixture to come to room temperature. The reactants are treated with T.L.C.
Tested with alcohol/chloroform, 1/10, v/v)
do. At the end of the reaction, the reaction mixture was poured into water (500ml).
I'll give it to you. Extract the product with chloroform (3
times, 250ml). The chloroform phase remains until the pH becomes neutral.
Wash with water and dry with sodium sulfate.
Concentrate. The obtained syrup (83g) is used as a compound as it is.
Used in manufacturing in 9a. Example 2 (Allyl 4-O-benzoyl-2,3-di-O
-benzyl-α-D-glucopyranoside) uro
acid (compound 8b) and the corresponding methyl ester
Production of compound 9b In the first step, allyl 4-O-benzoyl
-2,3-di-O-benzyl-6-O-triethy
-alpha-D-glucopyranoside (compound 6b) and
and allyl-4-O-benzoyl-2,3-di-O
-benzyl-α-D-glucopyranoside (compound
7b). 1. Production of compounds 6b and 7b Compound 6b is similar to that described for compound 6a
and get it. That is, in a pyridine solution of compound 5,
Next, add benzoyl chloride (1.5 equivalents) and react.
After t.l.c. (ethyl acetate/benzene, 1/20, v/v)
Put it on. Excess benzoyl chloride leads to excess methano
destroy it by adding a rule. Evaporation to dryness
After that, the residue was taken out again with chloroform and diluted with 10%
KHSO_FourWash with a solution of water, wash with water and dry.
Let it concentrate and dry. The syrup obtained is as it is.
It is used in the synthesis of compound 7b. This syrup
(105g, obtained using 30g of compound 3) was chloro
Dissolve in form (300ml). Paratoluence
Sulfonic acid (76 g monohydrate in 100 ml methanol)
Add. The next morning, the reaction is terminated (t.l.c., acetic acid
Ethyl/chloroform, 1/20, v/v). Black
Wash the loform phase with water until the pH is neutral and dry.
Dry and dry concentrate. The resulting syrup (98
g) on a silica gel (1.2Kg) column.
Graphite and elute with chloroform (0.6)
and then ethyl acetate/chloroform (1/20,
Elute with a mixture of v/v). In this way, pure
Obtain pure derivative 7b (30 g). This derivative 7b is
Used as is in the manufacturing step of compound 8b. Example 3 (Allyl-4-O-acetyl-2,3-di-O
-benzyl-α-D-glucopyranoside) uro
Production of methyl phosphate (compound 9a) The syrup obtained in the manufacturing step of compound 8a was
(300ml). Diazomethane A
At this time, add the solution until compound 8a disappears.
(t.l.c. ether/hexane, 1/1, v/
v). After acidification with acetic acid, the solvent is evaporated. Dissolve the resulting residue (53g) in hot ethanol.
let Derivative 9a crystallizes on cooling. recrystallization
After that, pure this compound 9a (18.4g) M.P.85-86
°C, [α]²⁰ _D= +12° (1,2 chloroform) obtained
Ru. This product was evaluated by its IR spectrum and N.M.R. spectrum.
Characterized by the vector and its elemental analysis. 7.6 g of compound 9a was also obtained from the recrystallization solution. The overall yield of compound 9a from compound 2 was 38%.
Ru. Example 4 (Prop-1'-enyl 4-O-acetyl-2,
3-di-O-benzyl-α-D-glucopyrano
Production of methyl uronate (compound 10a) Derivative 9a (4 g) was mixed with ethanol (119 ml).
Dissolved in a mixture of water (51ml) and water (17ml)
let Next, diazabicyclo (diazabicyclo)
Add cutane (170 mg) and bring to reflux. boiling solution
Tris(triphenylphosphine) chloride
Add Umu() (550mg). Continue boiling for 4 hours
(t.l.c., ether/hexane, 1/1,
v/v). At the end of the reaction, filter the solution to remove the solvent.
Ru. The residue was dissolved in ethyl acetate/chloroform (1/
50, v/v) to silica gel (150 g) in a mixture.
Separate by chromatography. Compound
Obtain 10a (3.25g; 81%). This compound 10a is
Crystallizes in tanol. [α]²⁰ _D= +12° (1, black
Roholm). M.P.90℃ structure was determined by elemental analysis and N.
Confirmed by M.R. and IR spectrum. Example 5 (Allyl-2,3-di-O-benzyl-α-D
-glucopyranoside)uronic acid and (allyl-
2,3-di-O-benzyl-α-D-glucopi
(ranoside) methyl uronate (compounds 11 and 12)
Manufacturing of Compound 8b (1.9 g) was dissolved in methanol (40 ml).
Let me understand. Next, add soda (5N) to a 1M concentration of soda.
Add only enough to give strength. After reaction T.L.C.
(methanol/chloroform, 1/4, v/v)
Put it on. When finishing the reaction, add water (100ml)
Add. Wash with ether, acidify, and then remove with ether.
Extract the product with tel. PH the acidic ether phase
Wash with water until it becomes neutral. Derivative 11 is isolated
It has not been. Derivative 11 of diazomethane
Methylation by a solution of
(900 mg; 56%). This compound 12 was added to silica gel.
in rum (ether/hexane, 1/1, v/v)
refine. [α]²⁰ _D=+35.2゜(1,3,chloroform
). Its IR and N.M.R. spectra and elements
Confirm the structure by analysis. In a similar way,
Derivative 11 is thus induced from compounds 9a to 9b.
It is possible to obtain a conductor 12. Example 6 Prop-1'-enyl 2,3-di-O-benzyl
-α-D-glucopyranosideuronic acid) methyl
Production of (compound 13) 1゜ When using compound 12 Derivative 12 was prepared as described for compound 9a,
Treatment with rhodium chain salts. Compound 13
Get a rate of 90%. This compound 13 has IR and N.M.R.
It is characterized by a spectrum. Furthermore, nothing
Treated with hydroacetic acid (1 ml for 180 mg of compound 9a)
to obtain compound 10a. 2゜ When using compound 10a or 10b Method a: Derivative 10a (350 mg) in methanol (5 ml)
Dissolve. Sodium methanolate (0.2ml,
2M). After 1 hour at ambient temperature,
dowex resine-50-H+
to stop the reaction. After evaporation, the product 13
obtain. Product 13 is a small amount resulting from 1′,αβ-elimination
contaminated by the products of reaction is compound
From 10b onwards, it is done in the same way. Method b: Derivative 13 is derived from compound 9a or 9b to compound 12.
obtained from compound 10a or 10b by the method described in
Is possible. Example 6A (see Figure 3) Synthesis of disaccharide 15 using compound 13 Monosaccharide 13 (0.215 g, 0.5 mmol) in dichloromethane
(3 ml) solution, monosaccharide 14 (0.49 g; 1 mmol)
Add chloromethane (3 ml) solution, then powder 4
Add a screen. After cooling the mixture to 0℃
Add sym-collidine (0.16ml) and add silver trifle.
(0.3g). After 1 hour, stir the mixture.
Dilute with lolomethane (50ml). Dehydrate the solid;
Solution: 5% sodium bicarbonate solution, 10% sulfuric acid solution
Wash with potassium and water in that order. Do it like this
After evaporation, 591 mg of residue is obtained. toluene/acetate
purified by silica in a 30/1 volume mixture.
After production, pure disaccharide 15 (211 mg) is obtained. Example 7 (see Figure 4) Bromo2,3-di-O-benzyl-4-O-a
Cetyl-α-D-glucopyranoside uronic acid
Production of chill (compound 25) Synthesis of compound 19 Compound 16 (32 g; 85.5 mmol) in pyridine (250
ml) solution, trityl chloride (28.6 g; 1.2 eq.)
Add and heat to 80℃. After 3 hours reaction, new
Add trityl chloride (4.6 g; 0.2 equivalents).
After compound 17 is completely formed (t.l.c. silica;
Tanol/chloroform, 1/20, v/v) solution
was cooled to 0°C, then benzoyl chloride (15 ml;
1.5 equivalents). The next morning, compound 18 was quantitatively detected.
It is formed. Then methanol (150
ml) dropwise and concentrated to dryness. the residue obtained
Contains para-toluenesulfonic acid (95g)
Remove into methanol (500ml). 2 hour reaction
After that, the reaction mixture was poured into a separating funnel containing ice water (2).
Move to Extract product 19 with chloroform
Use it as is for the next step. Infrared spectroscopy to purify a portion of this product
Analysis of the structure confirms the structure. colorless rubber-like
be. [α]²⁰ _D= -61° (chloroform). Synthesis of compound 23 Add the syrup (95g) obtained in the previous step to acetone (1
), then cooling the solution to 0 °C,
Add chromium oxide (52g) to sulfuric acid 3.5M (220ml)
Drop the solution. After 2 hours of reaction, the reaction mixture was
Pour into ice water (1). Product 20 in chloroform
(5 x 200ml). The pH of the chloroform phase
Wash until neutral, dry, and concentrate to dryness.
Ru. Dissolve the residue obtained above in methanol (650ml)
Then, add drops of sodium carbonate (20g) aqueous solution (50ml).
and heat the mixture to 50°C. The next morning, I got
The solution was partially concentrated and then dissolved in water (1.5)
Pour into. The aqueous phase was then washed with ether and acidified with hydrochloric acid.
After sexualization, product 21 is extracted with ether. Aete
Dry the phase with sodium sulfate and concentrate to dryness.
and obtained a yellow mass (50 g) containing compound 20.
Ru. This residue (50g) was mixed with acetic acid and trifluoroacetic acid.
(15/1, v/v, 615 ml)
let Stir this solution at 100℃ while adding water (160ml).
Add. The next morning, evaporate to dryness to remove trace amounts of acetic acid.
and evaporate the toluene. partially hydrolyzed
The residue consisting of compounds 21 and 22
(400ml). Add to this solution at 0°C until complete methylation.
Add ethereal solution of diazomethane (t.l.c.
ether/hexane, 2/1, v/v). past
Decompose excess diazomethane with acetic acid and react
The mixture is concentrated to dryness. The residue was first filtered through a silica gel (200 g) column.
Pure chloroform, then chloroform/ether
Eluted with a mixture of 3/1, v/v
refine. In this way, compound 23 (8.6 g; 22.2
mmol, 26% of compound 16). Derivative 23 crystallizes at M.P. 122-123°C. element
Structure confirmed by analysis and N.M.R. spectrum
It was done. Synthesis of compound 24 Compound 23 (3.9 g; 10 mmol) in pyridine (50
ml) Add acetic anhydride (4 ml, 42 mmol) to the solution.
Ta. After 2 hours the reaction mixture was evaporated to dryness. child
Compound 24 (4.62 g; 98%) is obtained as follows. Synthesis of compound 25 Compound 24 in dichloromethane (30 ml) and acetic acid ethyl
Titanium tetrabromide in chill (3 ml) solution (1.4 g)
(1.5g). Stir the solution overnight at room temperature.
After dilution with dichloromethane, the reaction mixture was placed in ice water.
pour. The organic phase was washed with 5% aqueous bicarbonate solution and dried.
Dry and concentrate. Chroma the residue on silica (50g)
(ether/hexane, 1/
1, v/v). Compound 25 (920 mg, 62%) was thus obtained
Ru. It is colorless and syrupy. [α]²⁰ _D=+97.5°
(C=1, chloroform). Elemental analysis and N.M.R.
The structure is confirmed by the spectrum. Example 7a (see Figure 5) Synthesis of disaccharide 27 using compound 25 This synthesis is carried out from derivatives 25 and 26. derivative 26
(432mg, 3mmol) dissolved in dichloromethane (10ml)
The liquid was mixed with 4Å molecular sieve (0.5g) and
dryerite (1g) and newly manufactured
Stir at 0 °C in the presence of silver carbonate (0.42 g).
Ru. After cooling at 0°C, compound 25 (490 mg, 1 mmol)
A dichloromethane (6 ml) solution of is added dropwise.
After 2 hours of reaction, the reaction mixture is filtered. evaporation
Dry and chromatograph the residue on silica gel.
When applied (solvent: ethyl acetate/chloroform,
1/6, v/v) derivative 27 is obtained. The structure of derivative 27 was determined by elemental analysis and N.M.R. spec.
Confirmed by Tor. Optical rotation: [α]²⁰ _D=
39°; Chloroform; M.P. 156-159°C. Example 8 (see Figure 6) (Trichloroacetimidyl 2,3-di-O-
Methyl benzyl-4-O-acetyl)uronic acid
Production of (compound 29) Synthesis of compound 28 Derivative 10a (11.4 g) Acetone/water mixture (5/
1; v/v; 350 ml). in this solution
Add mercury oxide (13.5g) followed by mercury chloride (17g)
Dropwise addition of an acetone/water mixture (116 ml) solution of
do. After 5 minutes, strain the mixture and evaporate the solvent.
let Soak the residue in chloroform (300ml)
and washed with a saturated solution of potassium iodide and then water.
Purify. After drying and evaporating to dryness, remove the residue from silica gel.
Column (200g, hexane/ethyl acetate, 1/
1, v/v). In this way the compound
28 (6 g; 57.5%) is obtained. It is solid. M.P.104
−106℃. [α]²⁰ _D=-4.5° (1,2 chloroform)
.
The structure was confirmed by N.M.R. spectrum and elemental analysis.
recognized. Synthesis of compound 29 Compound 28 (3.46 g; 8 mmol) and trichloro
- acetonitrile (8 ml; 10 eq.) dichloromethane
Sodium hydride (135 mg) in tan (80 ml) solution
Add. Check the progress of the reaction with T.L.C. (chloroform/vinegar
Check with ethyl acid, 1/1, v/v).
After filtration, the solution was concentrated to dryness and the residue was purified with benzene.
dried by evaporation and then reacted with compound 30.
Use accordingly. Example 8A (see Figure 6) Synthesis of disaccharide 31 using compound 29 Product 29 obtained above (8 mmol of compound 28
) containing derivative 30 (1.1 g; 7.6 mmol)
Dissolve in dichloromethane (40ml). This melt
Add dichloromethane of boron trifluoroethylate to the solution.
Add methane (0.6 eq.) solution dropwise. reaction
The progress of t.l.c. (chloroform/ethyl acetate, 6/
1, v/v). solid heavy carbon
Add sodium chloride until neutralized, then dichloromethane
Wash the phase with water until the pH is neutral. of solvent
After drying and evaporation, the resulting syrup is coated with silica gel.
Chromatograph (200g, chloroform/acetic acid
Ethyl, 8/1, v/v). in this way
Compound 31 (1.56 g) was obtained. Another anomer (α-D-glucuronosyl
(glucuronosyl)) is also obtained in small amounts (250 mg).
Compound 29 is the applicant's French patent application no.
Product 31 disclosed in No. 8200621 and in all respects
It is similar. Example 9 (see Figure 7) (trichloroacetimidyl-2,3-di-O
-benzyl-4-O-allyl-α-D-gluco
Preparation of methyl uronate (pyranoside) (compound 38)
Construction Synthesis of derivative 37 Allylation of compound 32 using conventional methods (natohydride)
Allyl bromide dissolved in dimethylformamide
solution) to obtain compound 33. For compound 33, here:
The same sequence of reactions described above for compound 18
Perform the reaction. (Detritylation, oxidation, hydrolysis,
methylation). At this stage, the product is silica gel
Rum (200g; chloroform/ethyl acetate; 10/
1; v/v). Compound 37 in syrup
Get it in shape. [α]²⁰ _D= +23.5° (1, chloroform
). Structure confirmed by infrared spectrum and elemental analysis
be done. Synthesis of compound 38 compound 37 (as described for compound 29)
trichloroacetate in the presence of sodium hydride
Treatment with nitrile gave imidate 38, which
Imidate 38 is used in the synthesis of compound 40. Example 9A (see Figure 7) Preparation of disaccharide 40 using compound 38 Synthesis of disaccharide 40 Compound 38 (615 mg; 1.1 mmol) and Compound 39
(300 mg; 1 mmol) in dichloromethane (5 ml)
Add boron trifluoro-ethyl to the solution at 20°C.
(1 mmol) in dichloromethane (10 ml)
Add liquid. The reaction was carried out using T.L.C. (acetone/toluene;
Check with 1/10; v/v). reaction mixture
Dilute with dichloromethane then sodium bicarbonate
(500mg) to neutralize. The resulting solution is diluted with sodium chloride.
Washed with a saturated solution of
Let it harden. Transfer the residue to a silica gel column (50 g;
Luene/isopropyl ether; 4/1; v/
v) chromatograph. In this way
Compound 40 (147mg; [α]²⁰ _D= +11°, chloroform)
and isomer (162 mg; [α]²⁰ _D=+48°, 1, chloro
holm). Elemental analysis and N.M.R. spectrum
Analysis of the sample confirms the structure of the product. The equations of Examples 10 to 12A are shown in FIG. Example 10 Prop-1-enyl-2,3-di-O-bendi
Ru-4-O-chloroacetyl-α-D-gluco
Production of methyl pyranoside uronate (compound 41) Compound 13 of Example 6, prop-1'-ethyl-
2,3-di-O-benzyl-α-D-glucopyra
Synthesized from methyl nosiduronate. Compound 13, 2.8g in pyridine 30ml (6.56mmol)
dissolve in it. After cooling to 0°C, chloroacetyl chloride
Drop 2 ml of dichloromethane (20 ml) solution
Add. After 30 minutes, evaporate to dryness and chloro
Extract with 200ml of holm and KOHSO_Four10% solution of water
Wash, dry, and concentrate in this order. The obtained white
The spout was soaked in silica gel (200 g; eluent AcOEt/hexane).
1/3; v/v) chromatograph
Ru. In this way, 2.7 g of pure compound 41 is
Obtained in the form of a lobe (yield 80%); [α]²⁰ _D=
+2゜(C= 1.5; chloroform). Elemental analysis and N.
M.R. spectrum confirms the desired structure. Example 11 2,3-di-O-benzyl-4-O-chloroa
Methyl cetyl-D-glucopyranose uronate
Production of (compound 42) 2.7 g (5.3 mmol) of derivative 41 in acetone/water
Dissolve in 80 ml of (5/1; v/v) mixture.
Add mercury oxide (3.1 g), followed by mercury chloride (3.9 g).
Add a solution of g) in acetone (27 ml). 5 minutes later,
Salt is removed by filtration. After concentration to dryness, the residue
Remove with chloroform. 10% chloroform phase
Wash with KI solution and then water. After evaporation, the product
is crystallized in an ethyl acetate/hexane mixture.
M.P.105−107℃; [α]²⁰ _D= −4.7° (equivalent, 1; chlorine
2 g of solid roform) are obtained. Elemental analysis and
The structure is confirmed by NMR analysis (yield 80
%). Example 12 Bromo-2,3-di-O-benzyl-4-O-
Chloroacetyl-α-D-glucopyranosyl
Production of methyl lonate (compound 43) 2g (4.30mmol) of compound 42 in dichloromethane
Dissolve in 50ml of water. sym-collidine 4.8ml (34.4
mmol) at 0°C, HEPBURN D.R. and
HUDSON H.R.J.Chem.Soc.Perkin I (1976)
Methylene dimethyl acetate prepared according to 754-757
Add ammonium bromide (17 mmol). 4 hours
After the reaction, the mixture was diluted with 100 ml of dichloromethane,
Then pour into ice water. After washing with ice water, evaporate the solvent.
let Silica gel (20g; eluent hexane/acetic acid
Ethyl, 2/1; v/v) chromatograph
Compound 43 was obtained in the form of 2.06 g of syrup.
(90% yield). [α]²⁰ _D=+82.5゜(C=1.5;
chloroform). By elemental analysis and N.M.R. analysis
The structure is confirmed. Example 12A Disaccharide 45, i.e. 3,0-acetic acid, using compound 43
Chil-1,6-anhydro-2-azido-4-
O-[(2,3-di-O-benzyl-4-O-k
Loloacetyl-β-D-glucopyranosyl)
Synthesis of methyl lonate β-D-glucopyranose
Growth 870 mg (3.8 mmol) of compound 44 in dichloromethane
Drierite (1g) and powdered 4Å
Molecular sieve (0.5g) and newly manufactured
Add silver carbonate (0.525g). Stir for 2 hours
After that, 670 mg (1.3 mmol) of compound 43 was added dropwise at 0°C.
and add. After 6 days, remove the solids by filtration.
Ru. The syrup obtained after concentration was immersed in silica gel (50%
g; Eluent: chloroform/ethyl acetate; 4/
1, v/v). Disaccharide 45
obtained in the form of foam (421 mg; 50%). [α]²⁰ _D
=-17゜(C=1; chloroform). By elemental analysis
Check the structure. Glycosi by N.M.R. analysis
Check the β-coordination of the bond between atoms. Example 13 (see Figure 9) formula has. 3-O-benzyl-α-L-glucopyranoside
Production of methyl iduronate (compound 56) Carry out this synthesis from compound 46 according to the following steps:
cormorant. That is, 1 Introduction of benzoyl group at position 5 2 Methylation of carboxyl functional group at position 6 3 Isomerization of OH group at position 5 4 Formation of pyran ring. 1 Benzoylation reaction 3-O-benzyl-1,2-O-isopropyly
Den-α-D-glucofuranoside (compound 46) 63
Dissolve g in 500 ml of anhydrous pyridine. Chloride
Add 85g of litil and heat to 80℃ for 1 hour. this
Compound 47 is thus obtained. Optical rotation: [α]²⁰ _D= -34.7°, chloroform. this
Infrared and N.M.R. spectra, original
Confirmed by elementary analysis. The mixture was then cooled to 0°C and benzoyl chloride 45
Add ml. The next morning, add 300ml of methanol and
Decompose excess reactants. The resulting mixture is
Evaporate to dryness and remove with chloroform. Chloroho
Wash the lume phase with water and dry with sodium sulfate.
and concentrate. Compound 48 is thus obtained. Dissolve the resulting syrup in 400ml of chloroform.
Let me understand. 5M para-toluenesulfonic acid methano
After adding 100ml of the solution, store the solution at 4℃ until the next morning.
put. After washing the organic phase with water, 215 g of the mixture
get. This mixture was mixed with silica gel (ether/
Chromatography with hexane, 2/1 (v/v) solvent
Apply it to the graph. In this way, 36g of compound 49
obtain. Optical rotation: [α]²⁰ _D= 65.3°, chloroform. transformation
The structure of compound 49 was determined by infrared and N.M.R. spectra.
I checked. 2 Methylation of carbonyl function at position 6 Compound 49 (1.88g) was dissolved in acetone (20ml).
Let me understand. CrO₃(13g) of 3.5MH₂S.O._Four(29ml)
Add 3.5 ml of the solution dropwise at -50°C. Leave it warm
Raise the temperature and leave in this state for 1 hour. next
Pour the reaction mixture into ice and remove the product from chloroform.
Extract with After washing with water, drying, and evaporating to dryness,
Compound 50 is obtained. The resulting mixture was dissolved in methanol (20 ml).
Then add 10ml of 1N soda and leave at room temperature until the next morning.
Leave it there. The reaction mixture was preliminarily diluted with methanol.
H washed with water⁺Shape of Dowex 50 resin color
(25ml). The product is concentrated by concentrating the eluate.
get it. Compound 51 is thus obtained. This compound is dissolved in ether and
Methylated with diazomethane according to the method. evaporation
After that, compound 52 (1.08 g; 70.4%) is obtained. optical rotation
[α]²⁰ _D=-27°chloroform. Based on elemental analysis, infrared and N.M.R. spectra
to confirm the structure of compound 52. 3 Isomerization of -OH group at position 5 triflic anhydride (0.8ml)
Cool the dichloromethane (16 ml) solution to -20°C.
Then, add pyridine (0.8ml) and dichloromethane to this.
(8 ml) solution dropwise. Then at -10℃
compound dissolved in dichloromethane (8 ml)
Add 800 mg of substance 52 dropwise. After 1 hour at -50℃,
The reaction mixture was added to a solution containing 160 g of sodium bicarbonate.
Pour into a mixture of water and ice (8 ml). with organic phase
Stir until the aqueous phase separates. 3% organic phase
HCl and H₂Washed with O and saturated NaCl solution
, dry and concentrate. Combined in this way
Get item 53. Remove the syrup again with DMF (10 ml). bird
Add sodium fluoroacetate (1.6g) and
Heat to 80 °C for an hour. In this way, compound 54 is
obtain. After evaporation, extract with dichloromethane and wash with water.
and dry, remove the residue with methanol,
The solvent is then evaporated after 1 hour. ether/heki
Column chromatography using San 2/1 as a solvent
Multiplying gives compound 55 (450 mg; 56.2
%). Optical rotation: [α]²⁰ _D= -33°, chloroform. The structure of compound 55 is infrared and N.M.R. spectra,
Confirm by elemental analysis. 4 Formation of pyran ring This synthesis is carried out from compound 55. Compound 55 (200
mg) in a mixture of trifluoroacetic acid/water (9/1)
Dissolve in. After 15 minutes, the solvent is evaporated. residual
The material is crystallized in ethyl acetate/hexane. this
In this way, 110 mg of compound 56 is obtained. The characteristics of this derivative are as follows. -Infrared spectrum: CHCl₃Medium cm^-1Unit γ: 3450
(OH), 3080, 3060, 3030 (CH₂: benzyl)
and 1740 (COOCH₃) −N.M.R. spectrum: ppm unit with TMS as standard
δ of place: 3.75 (S, 3H⁺, COOMe) 4.98 (1H⁺),
7.30 (S, 5H⁺,C₆H_Five) −Optical rotation [α]²⁰ _D=+13°, methanol −Elemental analysis C₁₄H₁₈O₇ Theoretical value Actual value C… 56.37 56.17 H…6.08 5.85 −M.P. 125−126℃ Example 14 (see Figure 10) Compounds 57,58 i.e. 1,2,4-tri-0-acetic acid
Thyl-3-0-benzyl-α,β-L-methyl
Production of idopyranuronate In French patent application no. 8200621 by the applicant
Therefore, the prepared compound 56 (3 g) was added to anhydrous pyridine.
(20ml) and acetic anhydride (10ml).
and stir at 0°C for 5 hours, protected from moisture. reaction
The mixture was evaporated to dryness and steamed with toluene (4 x 20ml).
Allow to air and dry under vacuum. Remove residue from silica gel
Chromatograph lamb (150 g). Tor
Dissolved in a mixture of ene:ethyl acetate (4:1 by volume).
When released, the following substances are obtained in sequence. - a first fraction consisting of a furanic acid derivative, - Compound 58 (anomer α), syrup (170 mg, 4
%), [α]_D=-43゜;(C:1, chloroform),
N.M.R. (CDCl)₃): δ: 6.23 (s, 1H, H-1). -Compound 57 (anomer-β), ether-hexanization
Crystallized into a compound, (2.688g, 63%), M.P.: 112−
113℃, [α]_D=+9゜(c.:1, chloroform),
N.M.R. (CDCl)₃): δ: 6.08 (d, 1H, H-1,
J_1,2:1.5Hz). When treated with the above synthesis process, the anomer becomes pregelatinized.
Compound 58 and anomeric β compound 57 are not separated. Use the mixture as a syrup for subsequent reactions.
used for. Example 15 (see Figure 10) 2,4-di-0-acetyl-3-0-benzyl
-β-L-methylidopyranuronyl bromide
Production of (compound 59) Acetate mixture of compounds 57 and 58 (212 mg, 0.5 m
M) in anhydrous dichloromethane (5 ml) and anhydrous acetic acid ethyl chloride.
Dissolve in chill (0.5ml). titanium tetrabromide
(250mg, 0.7mM) was added all at once to the reaction mixture.
Stir for 24 hours at room temperature, protected from moisture. to 0℃
After cooling and diluting with dichloromethane, the organic phase was poured into ice water.
Washed (3 times), dried (with sodium sulfate),
Filtration and evaporation give compound 59 (217
mg, 96%). N.M.R. (CDCl)₃): δ: 6.41
(s, 1H, H-1). This compound is extremely
Unstable, use immediately for subsequent reactions. The structural formulas of the compounds of Examples 16 to 19A are shown in Figure 10.
As shown in FIGS. Example 16 4-0-acetyl-3-0-benzyl-1,2
-0-methoxyethylidene-β-L-methyl
Production of dopyranuronate (compound 60) (0.425g, from 1mM acetate mixture 57+58
freshly prepared) anhydrous dichloromethane of bromide 59
(10 ml) solution at room temperature under a dry argon atmosphere.
Stir. sym-collidine (0.66ml, 5mM) and
and anhydrous methanol (0.4ml, 10mM) were added sequentially.
and stir the reaction mixture for 20 hours under the above conditions.
After dilution with dichloromethane (50ml), saturated bicarbonate
Wash the organic phase sequentially with aqueous sodium solution and water.
Cleaned, dried (sodium sulfate), filtered and evaporated.
emanate. Collect the residue on a silica gel column (20g).
Apply to matograph. (0.5% triethylamine
Hexane:ethyl acetate mixture in a volume ratio of 3:2 containing
When eluted with compound 60, compound 60 becomes pure syrup.
(302 mg, 76% of compound 57+58)
Ru. [α]_D=-21゜(c.:1, chloroform), N.M.
R. (CDCl₃): δ: 5.52 (d, 1H, H-1, J_1,2:3
Hz). Example 17 4-0-acetyl-3-0-benzyl-1,2
-0-tert-butoxyethylidene-α-L-methane
Production of tiloid pyranuronate (compound 61) (2.122g, from 5mM acetate mixture 57+58
freshly prepared) anhydrous dichloromethane of bromide 59
(20 ml) solution at room temperature under a dry argon atmosphere.
Stir. sym-collidine (2.65ml, 20mM) and
and anhydrous tert-butanol (3 ml, 30 mM).
and stir the reaction mixture for 15 h under the above conditions.
Ru. Treated in the same manner as compound 60 and purified the residue with silica gel.
Chromatograph on column (120 g).
(2:1 volume ratio with 0.5% triethylamine)
) When eluted with a hexane:ethyl acetate mixture
Compound 61 is pure syrup (1.542 mg, Compound 57 + 58
70%) of the shape. [α]_D=-23゜(c.:
1, Chloroform), N.M.R. (CDCl₃): δ: 5.48
(d, 1H, H-1, J_1,2:2.5Hz). Example 18 3-0-benzyl-1,2-tert-butoxye
Tylidene-α-L-methylidopyranuronate
Production of (compound 62) Anhydrous solution of orthoester 61 (484mg, 1.1mM)
Stir the tanol (15 ml) solution in a dry argon atmosphere.
Cool to −20° C. under stirring. Anhydrous potassium carbonate (60
mg) and stirred the reaction mixture for 5 hours under the above conditions.
Stir. Dehydrate the solid, evaporate the liquid and remove the residue.
Place in chloroform (50ml). Organic phase in ice water
Washed (3 times), dried with (sodium sulfate),
Filter and evaporate. Immediately remove the residue using silica gel.
(25 g). (0.5%
(2:1 volume ratio) hexa containing triethylamine
Elution with an ethyl acetate mixture yields the following:
Quality is obtained sequentially. -Unsaturated compound 64 (31 mg, 7%) syrup, [α]_D
=+103゜(c.:1, chloroform), N.M.R.
(CDCl₃): δ: 6.27 (d.d., 1H, H-4, J_3,4:
5Hz, J_2,4: 1Hz), 5.67(d, 1H, H-1,
J_1,2:4Hz). -Main fraction (271 mg, 62%). This fraction is ether-
Crystallizes in hexane mixture to form compound 62
(123mg, 28%), M.P.: 68-69℃; [α]_D=-
19゜(c.:1, chloroform), N.M.R.
(CDCl₃): δ: 5.41 (d.1H, H-1, J_1,2:2
Hz), 2.85 (d, 1H, CH-4, J: 12Hz, D₂O
exchange). During chromatography using silica gel column
and during the crystallization process of compound 62, slightly more than compound 13.
New compounds with high Rf are produced. transformation
Silica gel chromatography of crystallized mother liquor of compound 62
The pure fraction of this new compound 65 is
Some can be isolated. (41 mg, 11%), syrup,
[α]_D=+21゜(c.:1, chloroform), N.M.R.
(CDCl₃): δ: 5.83 (d.1H, H-1, J_1,2:4.5Hz). In the synthesis process of the present invention, the form of compound 65 is
The crude syrup 62 was chromatographed to prevent
Use directly for subsequent reactions without roughening. Example 19 3-0-benzyl-4-0-chloroacetyl-
1,2-0-tert-butoxyethylidene-α-
L-methyloid pyranuronate (compound 63)
manufacturing Anhydrous solution of orthoester 62 (220mg, 0.5mM)
Stir the tanol (10ml) solution in a dry argon atmosphere.
Cool to −20° C. under stirring. Anhydrous potassium carbonate (40
mg) and stirred the reaction mixture for 5 hours under the above conditions.
Stir. Drain the solid, evaporate the liquid, and remove the residue.
Place in chloroform (50ml). Immediately remove the organic phase
Wash with ice water (3 times) and then with (sodium sulfate).
Dry, filter and evaporate. Immediately remove the residue with anhydrous pipette.
Lysine (4 ml) and anhydrous dichloromethane (2 ml)
Dissolve in. -20℃ under dry argon atmosphere
After cooling to (0.1 ml freshly distilled, 1.25 mM)
Chloroacetyl chloride in anhydrous dichloromethane (1
ml) solution dropwise. The reaction mixture under the above conditions
Stir for 30 minutes, then pour into water-ice mixture (100 ml).
ingredient. After stirring for 15 minutes, the mixture was dissolved in chloroform (3x
Extract with 20ml). Ice water, 2% sodium bicarbonate
washing the organic phase sequentially with an aqueous solution and water;
Dry (over sodium sulfate), filter and evaporate.
Immediately transfer the residue to a silica gel column (12 g).
Apply to matograph. (0.2% triethylamine
Hexane:ethyl acetate mixture in a volume ratio of 5:2 containing
When the compound is eluted, the following substances are obtained in sequence. - unsaturated compound 64 (15 mg, 8%), - Orthoester 63 syrup (145 mg, Compound 62
61%) [α]_D=+19゜(c.:1, Chlorophor
), N.M.R. (CDCl₃): δ: 5.45 (d.1H, H-
1, J_1.2:2.5Hz), 5.24(d.d., 1H, H-4,
J_3.4:2.5Hz, J_4.5:1.5Hz), 4.00(s, 2H, Cl−
_CH2 −COO−). Example 19A (see Figure 12) Synthesis of disaccharide 67 using compound 63 Disaccharide 67 is the compound: Methyl-6-0-benzoyl-3-0-benzyl-2-benzyloxycarbonylamino-2-
Deoxy-4-0-(2,4-di-0-acetyl-3-0-benzyl-α-L-methylidopyranuronyl)-α-D-glucopyranoside. A solution of orthoester 63 (80 mg, 0.2 mM) and alcohol 66 (52 mg, 0.1 mM) in anhydrous chlorobenzene (8 ml) is heated to 140° C. with stirring in the presence of a slight stream of dry argon. After 6 ml of solvent has slowly distilled, (NKKOCHETKOV,
AFBOCHKOV，TASOKOLCVSKAIA et
VJSNIATKOVA, Carbonhdr.Res., 16 (1971)
17-27, freshly prepared according to 0.002mM)
A solution of 2,6-dimethylpyridium perchlorate in chlorobenzene (2 ml) is added dropwise over 15 minutes. Simultaneously with the addition (2 ml) of the solvent is distilled off. The reaction mixture is then stirred for 1 hour under the above conditions with simultaneous addition of fresh solvent (10 ml) and distillation so that a constant reaction volume equal to 2 ml is maintained. After cooling and diluting with chloroform, the organic phase was washed successively with saturated sodium bicarbonate solution and water,
Dry (over sodium sulfate), filter and evaporate.
The residue is chromatographed on a silica gel column (15 g). Elution with a hexane:ethyl acetate mixture (4:3 by volume) gives the following materials in sequence: - Starting material 66 (20 mg, 38%) - Homogeneous fraction by thin layer chromatography (54 mg).
According to the NMR spectrum, this fraction has multiple 0-methyl signals (δ: 3.35-3.50), and these signals are due to methyl glycosides generated by rearrangement of orthoester 63. It is something. This fraction crystallizes in an ethanol-water mixture and recrystallizes in an ethyl acetate-hexane mixture to yield compound 67 (44 mg, 50%).
MP: 120-121°C, [α] _D = +17° ( c : 1, chloroform), NMR (CDCl ₃ ): consistent with the desired structure. Example 20 (See Figure 13) Preparation of methyl-(methyl-2,3-di-0-benzyl-α-L-glucopyranoside)tinuronate (compound 79) The synthesis of this compound is carried out in the following seven steps. Step 1: Synthesis of monosaccharide 69 Compound 69 was synthesized in the Canadian Journal of Chemistry.
NL from Chemistry, 51 (1973), page 3357.
It is prepared from compound 68 obtained by the method of Holder and B. Fraser-Reid. Compound 68 (1g,
12.67mM) in dichloromethane (20ml),
Tosyl chloride (0.55 g), dimethylaminopyridine (16 mg) and triethylamine (0.7 ml) are added sequentially. After stirring for about 14 hours under nitrogen flow with exclusion of moisture, the reaction is stopped by adding ice and water. The reaction mixture is diluted with dichloromethane (50 ml) and the dichloromethane phase is washed successively with 2M hydrochloric acid, saturated sodium bicarbonate solution and finally with water until Hz becomes neutral. After drying and evaporation the residue or derivative 69
(1.4g, 97%) is obtained. Derivative 70 is synthesized using this derivative as it is. Step 2: Synthesis of derivative 70 Monosaccharide 69 (31.8g) and sodium iodide (39g)
and is dissolved in acetonitrile (250 ml) and the solution is refluxed for 3 hours. After cooling the reaction mixture, filter the white precipitate formed. Concentrate the liquid,
The residue is taken up in chloroform, then the chloroform phase is washed with water to neutral Hz, dried over sodium sulfate and concentrated to dryness. Silica gel column (200g,
A syrup is obtained by chromatography in an ether-hexane volume ratio of 1:1. This gives iodinated derivative 70 (24.7 g, 71.5%). [α] ^D ₂₀
= 24° (1, chloroform). The structure of compound 70 is confirmed by infrared spectrum, NMR spectrum, and elemental analysis. Step 3: Synthesis of derivative 71 To a solution of derivative 70 in anhydrous pyridine (200 ml) is added acetic anhydride (43 ml). The reaction is complete after about 14 hours of stirring. The reaction mixture is concentrated to dryness and the residue is purified on a silica gel column under pressure using ethyl acetate-hexane (1:6 by volume) as solvent. Collect multiple pure fractions into one. This gives product 71 (16.4 g, 70%). The product is syrupy. [α] ^D ₂₀ = +4.5° (1, 3,
chloroform), elemental analysis and infrared spectral analysis confirm the structure. Step 4: Synthesis of derivative 72 Silver fluoride (AgF, 6.9 g) is added to a solution of derivative 71 (4 g) in pyridine (100 ml) cooled to 0°C. After 2.5 hours, the reaction mixture is poured into a chloroform-ether mixture (1) (1:4 by volume). The resulting suspension is passed through a pleated filter. Concentrate the liquid to dryness and put the residue in chloroform (500 ml). The chloroform phase is washed with 10% aqueous potassium sulfate and then with water until neutral PH. The residue (2.7 g) obtained by drying with sodium sulfate and concentrating to dryness was applied to a silica gel column (200
Chromatograph g) (eluent: ethyl acetate-hexane in a volume ratio of 1:4). The fractions containing product 72 were combined and the solvent was evaporated to give a crystalline material (1.62 g, 54%) of M.
P.: 81-82°C, [α] ^D ₂₅ =-20° (1, chloroform). Infrared spectrum analysis, elemental analysis and
Analysis of the NMR spectrum confirms the structure of compound 72. Step 5: Synthesis of derivative 73 Product 72 (2 g) is dissolved in methanol (20 ml) and chloroform (20 ml). Add sodium methanolate (2M, 2 ml) to this solution.
After 1.5 hours, the deacetylation reaction is complete. Dilute the reaction mixture with chloroform. The chloroform phase is washed with water to neutral Hz, dried and evaporated to dryness. This leaves compound 73 (1.8
g, 100%). This residue is immediately dissolved in tetrahydrofuran (50 ml) and then 10 ml of a solution of borohydride (BH ₃ , 1M) in tetrahydrofuran is added. After 1 hour of reaction, excess borohydride is decomposed by adding ethanol. After the end of gas evolution, the reaction mixture is diluted by adding tetrahydrofuran (100 ml). Next, 3M sodium hydroxide (soda, 12 ml) and hydrogen peroxide (8 ml) (120 times the volume) are added sequentially. After heating at 50°C for 2 hours, the reaction is stopped. Add the solution to chloroform (5000
ml), the resulting organic phase was washed with water, then 2M
Wash with hydrochloric acid and finally with water until neutral Hz. This results in an extremely cloudy chloroform phase. This phase becomes transparent upon drying with sodium sulfate. After filtration, the chloroform is evaporated and the resulting residue is chromatographed on silica gel (200 g) (chloroform-methanol volume ratio 30:1). Idose derivative 73 (1.05g,
55%). The product is syrupy.
[α] ^D ₂₀ = +85.5° (1, chloroform). Elemental and NMR analyzes confirm the desired structure. Step 6: Synthesis of derivative 76 This synthesis (without isolating intermediates 74 and 75)
This is done in one step starting from derivative 73. Derivative 73 (2.25g, 6mM) in dichloromethane (50ml)
Add dimethylaminopyridine (60 mg, 0.24 m
M), triethylamine (1.7ml, 12mM) and trityl chloride (2.5g, 9mM) are added sequentially.
The reaction ends after about 14 hours. This results in a derivative
A solution of 74 is obtained. Now add dimethylaminopyridine (150 mg), triethylamine (1.7 ml) and benzoyl chloride (1.05 ml) to the reaction mixture. After 6 days, the dichloromethane is removed through a stream of nitrogen and replaced with dimethylformamide (40 ml). The reaction mixture is heated to 70°C and maintained overnight.
Here, benzoyl chloride (1 ml) and triethylamine (1.7 ml) were added again, heated to 70°C, and
Maintain for days. The dimethylformamide is then evaporated, the residue is taken up in chloroform and the chloroform phase is washed successively with water, saturated sodium bicarbonate solution, 2M hydrochloric acid solution and finally with water until neutral Hz. After drying, evaporation of the chloroform yields compound 75. Derivative 76 is obtained by immediate reaction to remove the trityl group from this compound. The residue containing derivative 76 was dissolved in 25 ml of chloroform, and the solution was added with methanol (1M) of paratoluenesulfonic acid-hydrate.
Add 10ml of solution. The reaction is completed after 4 hours of reaction at room temperature. The reaction mixture is now diluted with chloroform, washed with water, dried and evaporated to dryness. The residue obtained is chromatographed on silica gel (200 g, ether-hexane volume ratio 3:1). Derivative 76 (1.5g: 52%) in pure state through this process
is obtained. This derivative is syrupy.
[α] ^D ₂₀ = −8° (1, chloroform). Analysis of the infrared and NMR spectra confirms the structure of the desired product. Step 7: Synthesis of Compound 79 This synthesis is carried out directly from derivative 76 without isolating intermediates 77 and 78. After cooling a solution of Compound 76 (1.2 g) in acetone (20 ml) to 0° C., a solution (2.9 ml) of chromium oxide (CrO ₃ :1.17 g) in 3.5 M sulfuric acid (5 ml) was added dropwise. After stirring at 0°C for 30 minutes, bring to room temperature. The reaction lasts for 3 hours. Next, ice water (100
Pour the reaction mixture into a separatory funnel containing 1.0 ml of The product formed is extracted with chloroform (3 x 50 ml). The chloroform phase is washed with water until neutral Hz, dried over sodium sulfate, filtered and concentrated to dryness. The resulting residue (compound 77) is dissolved in methanol (130 ml). To this solution is added 3M sodium hydroxide (soda) solution (17 ml) and the mixture is kept under stirring for about 14 hours. Acidification with sulfuric acid and extraction of compound 78 with ether followed by immediate methylation with diazomethane provides compound 79 by conventional methods. After evaporation of the ether, chromatography on silica gel (50 g, ether-hexane; 4:1 volume ratio) yields pure compound 79. The pure fractions containing derivative 79 are combined and the solvent is removed. In this process, iduronic acid derivative 79 (587 mg,
59% of derivative 76) is obtained. The product is syrupy. [α] ^D ₂₅ = +98° (2.65, chloroform). The desired structure is confirmed by NMR analysis, infrared analysis and elemental analysis. Example 20A (See Figure 14) Synthesis of Disaccharide 81 Using Compound 79 This synthesis combines monosaccharide 79 prepared as described above.
chemische Berichte 111 (1978) 2234-2247, prepared by the method of H. Paulsen and W. Stenzel. A solution of compound 79 (220 mg, 0.5 mM) in dichloromethane (10 ml) was added with compound 80 (0.450 g), sym-collidine (150 μ), and silver triflate (tflflate).
d'argent) (260 mg). The reaction mixture is kept under stirring at 0° C. for 3 hours, protected from moisture and light and under nitrogen flow. The reaction mixture is then diluted with dichloromethane (100 ml) and filtered through a pleated filter to remove solids. The resulting solution was washed successively with saturated sodium bicarbonate solution, water and 2M sulfuric acid, and finally neutralized.
Rinse again with water until it reaches Hz. The residue obtained after drying over sodium sulfate and evaporation of dichloromethane is chromatographed on silica gel (50 g, chloroform/ethyl acetate; 15:1 by volume). This process produced pure derivative 81 (327mg, 82%)
is obtained. The product is syrupy. [α] ^D ₂₀
= +57° (1, chloroform). NMR and elemental analyzes confirm the structure and anome'rie of disaccharide 81. Example 21 Formula Synthesis of trisaccharide 86 This synthesis is carried out in three steps (first
(See Figure 5). First, glycosylation of orthoester of L-iduronic acid derivative is performed. Next, the monochloroacetyl group is selectively removed. Finally, one of the alcohols formed is reacted with a disaccharide. 1 Orthoester with benzyl alcohol63
Glycosylation of orthoester 63 (118 mg,
(0.25mM) and (freshly distilled) benzyl alcohol (0.15ml, 1.5mM) in anhydrous chlorobenzene (10ml) and heated to 140°C, protected from moisture. Slowly evaporate the solvent (8 ml) and then
2.6-dimethylpyridium perchlorate (2.5μM)
A solution of chlorobenzene (2 ml) was added to the solvent (2 ml).
was added dropwise over 30 minutes while distilling simultaneously. The reaction mixture is stirred for 30 minutes with simultaneous addition of fresh solvent and distillation of the solvent so that the conditions described above are maintained and a constant reaction volume equal to about 2 ml is maintained. After cooling and diluting with chloroform (50 ml), the organic phase is washed successively with 5% aqueous sodium bicarbonate solution and water, dried (over sodium sulfate), filtered and evaporated. The residue is chromatographed on a silica gel column (8 g). When eluted with a mixture of hexane-ethyl acetate (2:1 by volume), the glycoside α
Fraction containing mixture of and β82 (102 mg, 81%)
get. At this stage, glycosides α and β are not separated. NMR (90MHz, CDCl ₃ ): δ: 7.30
(m, 10H, 2Ph), 3.98 (s, 2H, Cl−CH ₂ −
CO), 3.74 (s, 3H, CCO Me ), 2.08 and 2.03
(2s, total 3H, O Ac forms β and α; β: α
and β2:1). 2 Selective 0-demonochloroacetylation Pyridine (5
ml) and absolute ethanol (1 ml) is heated at 100° C. for 20 minutes in the presence of thiourea (25 mg). After cooling, the reaction mixture is evaporated to dryness and the residue is taken up in a water-chloroform mixture (50 ml, 1:1 by volume). The organic phase is washed with water, dried (sodium sulfate), filtered and evaporated. The residue is chromatographed on a silica gel column (10 g). Elution with an ethyl acetate-hexane mixture (4:3 by volume) sequentially isolates the following substances: - Glycoside β83 (26 mg, 25%), colorless syrup,
[α] _D +70° ( C 1, chloroform), NMR
(90MPH, CDCl ₃ ): δ: 7.30 (m, 10H, 2
Ph); 5.05 (m, 1H, H ₂ ); 4.90 (d, 1H, H ₁ ,
1.2J=2Hz); 3.78 (s, 3H, COO Me ); 3.12
(1H, O H , D ₂ O exchange); 2.05 (s, 3H, O
Ac). - Glycoside α84 (54 mg, 50 of orthoester 63)
%), colorless syrup, [α] _D −65° ( c 1, chloroform), NMR (90 MPH, CDCl ₃ ): δ: 7.30
(m, 10H, 2 Ph ); 5.05 (2H, H ₁ and H ₂ , J _1,2
The coupling constant for is extremely small (1Hz), 3.78
(s3H, COOMe ); 2.80 (1H, OH , _D2O exchange), 2.06 (s, 3H, OAc ). 3 Glycosylation of alcohol 84 with disaccharide 85 Alcohol 84 (22 mg, 50 μM) and bromide 85 (57
anhydrous dichloromethane (1.5 mg, 70 μM)
ml) The solution is stirred in the presence of molecular sieves 4 Å (powder, 50 mg) protected from moisture and light.
The reaction mixture was cooled to -20°C and treated with sym-collidine (110μ) and silver triflate (26mg, 100μM).
are added sequentially. The reaction mixture was stirred under the above conditions for 2 hours, diluted with dichloromethane (50 ml), the solid was dried, and the solution was washed sequentially with cold 0.1 M HCl, water, 5% aqueous sodium bicarbonate, water, and diluted with dichloromethane (50 ml). ) Dry, filter and evaporate. The residue is chromatographed on a silica gel column (8 g, 230-400 mesh gel). Elution with a toluene-ethyl acetate mixture (5:1 by volume) allows the trisaccharide 86 to be isolated in the form of a colorless syrup (50 mg, 86%). The NMR spectrum (270MHz, _CDCl3 ) is consistent with the desired structure. Example 22 (see Figure 16) Formula A formula useful as an enzyme substrate using the L-iduronic acid derivative 87 shown in as well as Synthesis of Compound 89 of the Glycoside Compound 87 (French Patent Application No. 8201575:900
mg: 2mM) in dichloromethane (20ml). Here tributylstannylphenol
Stannyl Phenol (compound 88, 2.5 μM) and tin tetrachloride (2.5 mM) are added sequentially. After 5 hours, the reaction mixture is diluted with dichloromethane and washed successively with sodium bicarbonate solution and water. The residue obtained after drying and solvent evaporation (1.03 g) is directly hydrogenated. For this, the residue was dissolved in methanol and then Pd/C (5%; 0.5 g) was dissolved under hydrogen atmosphere.
The solution is stirred for 3 hours in the presence of . The catalyst is removed by filtration and the solution is concentrated to dryness. Acetylation of the residue with acetic anhydride in the presence of pyridine affords compound 89 (420 mg). The obtained compound 89 is purified on a silica gel column (100 g, ester/hexane; volume ratio 3:1). NMR spectrum and elemental analysis confirm the structure of compound 89. Synthesis of compounds 93 and 95 In a solution of compound 89 (400 mg) in glacial acetic acid (2.5 ml),
A mixture of acetic anhydride and nitric acid (1 ml, volume ratio 16:
5) was added dropwise at 0°C while stirring. Maintained at 35° C. for 2 hours, the reaction mixture is poured into ice water and the products formed (90 and 91) are extracted with chloroform. The chloroform phase is washed with 5% bicarbonate of water, dried and concentrated to dryness. Products 90 and 91 are formed in a ratio of approximately 2:1. These products were mixed with silica gel (25
chromatograph with ether/hexane (volume ratio 1:1). This process yields pure compound 90 (23 mg), pure compound 91 (19 mg), and a fraction containing both unseparated compounds 90 and 91. Compounds 92 and 94 are dissolved in methanol at 0°C. Make 4N sodium hydroxide (soda) basic
Drip until the pH is reached. Once the hydrolysis is complete, resin Dowex 50H+ is added to deionize the solution. Compounds 93 (11 mg) and 95 (10
mg). NMR spectra confirm the structures of these compounds. It is found that various substituents can be introduced to glycoside 89 after the enrichment step. Protecting groups can be removed all together or sequentially, e.g. from the 2 and/or 3 and/or 4 positions, to produce special substituents, e.g. various glycosides.
SO ₃ groups can be introduced. Other glycosides were also prepared using L-iduronic acid and D-glucuronic acid with substituents different from those of derivative 89.

[Brief explanation of drawings]

1 to 16 are process diagrams for producing the compound of the present invention. A is an allyl group (including those via a -C ₆ H ₅ group), Bn is a benzyl group, Tr is a trityl group, R ₆ is an -OAc or -OBz group (however, Ac is an acetyl group, Bz
is a benzoyl group), Me is a methyl group, P is a vinyl group, MCA is a monochloroacetyl group, Tf is a trifuryl group, t-Bu is a quaternary butyl group, and SnBu ₃ is a tributyl stannyl group.

Claims (1)

[Claims] 1. A derivative having a uronic acid structure, which has the formula (): [In the formula, R ₁ is a halide, an O-imidoyl group, an adjacent
OR orthoester group formed with ₂ substituents, O
- represents a reactive residue selected from alkenyl groups and substituted or unsubstituted alkyl or acyl groups and free OH groups or blocked by protecting groups selected from alkenyl groups; (However, when R ₁ represents a methoxy group, OR ₄ is not an alkoxy group), OR ₂ , OR ₃ and OR ₄ are O-alkenyl groups,
or represents a free OH group or blocked by a protecting group defined for R ₁ (with the exception of OR ₂ in the case of glucuronic acid derivatives);
OR ₃ and OR ₄ are not all the same;
When R ₁ and OR ₂ are taken together to form an orthoester, OR ₃ and OR ₄ are different from acetyl groups, and R ₁ represents a halogen atom and OR ₄
When represents a methoxy group, or when R ₁ represents an O-alkyl group and OR ₄ is an OH group, OR ₂
and OR ₃ is different from a benzyl group), R ₅ is a hydrogen atom or a protecting group as defined above], or salts of these derivatives (provided that R ₁ , OR ₂ and OR ₃ are all O-benzyl group and R ₅ represents a methyl group), excluding iduronic acid derivatives. 2 Formula (): (wherein the substituents have the meanings defined for formula (), provided that R ₅ does not represent a hydrogen atom) derivative. 3 Formula (): (wherein all substituents have the same meanings as defined for formula ()). 4 R ₁ and OR ₂ represent orthoester,
Alternatively, the derivative according to claim 3, wherein R ₁ is a halide, an alkoxy group, or an O-alkenyl group. 5 It has a uronic acid structure and has the formula (): [In the formula, R ₁ is a halide, an O-imidoyl group, an adjacent
OR orthoester group formed with ₂ substituents, O
- represents a reactive residue selected from alkenyl groups and substituted or unsubstituted alkyl or acyl groups and free OH groups or blocked by protecting groups selected from alkenyl groups; (However, when R ₁ represents a methoxy group, OR ₄ is not an alkoxy group), OR ₂ , OR ₃ and OR ₄ are O-alkenyl groups,
or represents a free OH group or blocked by a protecting group defined for R ₁ (however, in the case of glucuronic acid derivatives, OR ₂ , OR ₃
and OR ₄ are not all the same and R ₁
and OR ₂ together form an orthoester, OR ₃ and OR ₄ are different from acetyl groups, R ₁ represents a halogen atom and OR ₄ represents a methoxy group, or R ₁ is O- (represents an alkyl group and OR ₄ is an OH group, OR ₂ and OR ₃ are different from a benzyl group), R ₅ represents a hydrogen atom, or a protecting group as defined above], or derivatives corresponding to these derivatives. A method for producing a salt (excluding iduronic acid derivatives in which R ₁ , OR ₂ and OR ₃ all represent an O-benzyl group and R ₅ represents a methyl group), comprising the formula (): [In the formula, R ₇ to _{R 9} may be the same or different and represent the above-mentioned protecting group for blocking a hydroxyl group] at the 6-position. selectively blocks the -OH group of -CHOH at position 5, selectively blocks -OH group of -CHOH at position 6, and selectively deblocks -OH group at position 6 to form -
Obtain a CH ₂ OH group, oxidize the -CH ₂ OH group to a -COOH group, and convert the 5-position -
The OH group is deblocked and, if desired, the -
The COOH group is converted to an ester group, a reactive residue is introduced at the 5-position, the configuration of the carbon atom at the 5-position is changed, the reactive residue at the 5-position is removed, and the nodofuranoside ring is rearranged. A method characterized in that it consists of treating the idopyranosidic ring. 6. A corresponding α-D-glucofuranoside derivative corresponding to the formula (
selectively blocks the -CHOH group at the 5-position.
By selectively blocking the OH group with a benzoyl group and selectively unblocking the -OH group at the 6-position, -
A CH ₂ OH group is obtained, the -CH ₂ OH group is oxidized to a -COOH group using chromic oxide as an oxidizing agent, the -OH group at position 5 is deblocked by treatment with soda, and, if desired, The -COOH group at the 6-position is converted to a -COOMe group using diazomethane as an esterifying agent, and a reactive residue selected from tosylate, mesylate, and trifuryl groups is introduced at the 5-position to change the configuration of the carbon at the 5-position. is changed using a nucleophile, the functional group at the 5-position is removed using methanol, and then purified by chromatography, and the ring is rearranged in an acidic medium to change from an idofuranoside ring to an idopyranoside ring. 6. A method according to claim 5, characterized in that the method comprises: processing.