JPH02138290A - Novel immnoadjuvant compound and its manufacture - Google Patents

Abstract

NEW MATERIAL: Compounds of formula I (R and R¹ are each H, acetyl, octanoyl, stearoyl, beenoyl or benzoyl; R² and R⁶ are methyl; X is L-valyl or L-phenylanyl; Y is D-isoglutamine; wave lines is α- or β-coordination or their mixture, when one of the wave lines is α, the other is β; when R and R¹ are H, X is not L-valyl).

EXAMPLE: 2-(4,6-Di-O-acetyl-2-acetamide-2-deoxy-D-glucose-3-O-yl)-D-propionyl-L- valyl-D-isoglutamine.

USE: Immunoadjuvants.

PROCESS: A compound givent by formula II (R', R¹', R²' and X' are R, R¹, R² or X respectively; Z and P are H; at least one of R', R¹', R²', X', Z and P is a protective group) is catalytically hydrocracked.

COPYRIGHT: (C)1990,JPO

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to hydrogen, acetyl, octanoyl, stearoyl, behenoyl or benzoyl; R2 represents methyl; R6 represents methyl; X represents L-valyl or L
- represents phenylalanyl; Y represents on D-isoglutamine; and the wavy line (te) represents α- or β-configuration, or also a mixture thereof fir: where the wavy line 10,000 is α In the case, the other is β; however, R and R
If 1 represents hydrogen, then X cannot be L-valyl.

In the reaction diagrams described below, for the sake of brevity and to avoid excessive redundancy, reference will be made to novel immunoadjuvant compounds represented by formula 1 above.

In the above formula 1, R and R1 are the same and represent the part of the formula corresponding to the part shown by
It is understood that it has the configuration structure of the subformula indicated by 0.

Particularly preferred are compounds of formula 1 in which X is L-valyl and Y is D-isoglutamine.

In the field of immunology, in order to maintain a sufficient host immunological response to prevent viral (or bacterial) infection,
More than one type of vaccine (or bacterin) is usually required. This means that the viral (or bacterial) antigens contained in the vaccine (or bacterin) are cleared very rapidly from the site of injection by the animal (or human) host, leaving insufficient time for the host to mount its immune response. This is because you can only have it.

To delay the release of viral (or bacterial) antigens and also to generally stimulate the host's immunological system, an immune adjuvant is added to the vaccine (or bacterin) and the injections are administered multiple times. However, efforts are being made to eliminate the need for these traditional immune adjuvants, such as kaolin, gum tragacanth,
bentonite, carbobol, calcium phosphate,
Many of tapioca, alum, aluminum hydroxide, calcium chloride, sodium alginate, etc. are non-metabolizable when injected and therefore have the disadvantage that these immune adjuvants themselves are irritating sources.

There has long been a need in the art for immunoadjuvant substances that can be easily metabolized by the host, while simultaneously slowing the release of antigen and generally stimulating its immune response.

An immune adjuvant commonly used in the laboratory is AQcobacterium tuberculosis.
Freund's complete adjuvant (FCA), which is a suspension of killed bacteria of sis) in mineral oil and an emulsifier; and (1) Freund's incomplete adjuvant (FCA), which is only mineral oil and an emulsifier. PC'A and? Although A is used as a laboratory standard, they are not used commercially. This is because PCA is derived from the toxic microorganism, Mycobacterium tuberculosis; and because FA does not produce a sufficient immunological response in relation to VC in the absence of killed Mycobacterium tuberculosis.

Ellouz at al is a member of Biochemical and Biophysical Research Communications.
iophysicalResearch Com! nu
nications), Vol. 59, No. 4, 161)-13
25 (1974) describes that peptidoglycan species obtained from microorganisms have immunoadjuvant activity and can be used in place of killed Mycobacterium tuberculosis in FCA (also, Belgian Special B!T). - see also No. 821,385].

The novel peptidoglycans of formula (I) can be used to improve antibody titres against either bovine serum albumin (BSA) or ovalbumin, for example.
) has significant immunoadjuvant activity as determined by W.G. Harbor (W, J.).

1'1erbert), Ch, 20, Volume 1, Handbook of Experimental Immunology.
tal Engineering), Editor W.M.Ware (Fiditor)
D, M, Wθ1r), Blackwell Scientific Hook Replications, Oxford (B
rackwell 8cientific Public
cations, Oxford) (1973)], and as detailed in Example 16, the BOA
delayed-type hypersensitivity to either ovalbumin or ovalbumin [G.H. Ham 7Elly and R.G.
White (J.

H, Humphray and R, G, White
) * Immunology for 8tu
dents of Mediana).

493-545M; Publisher F.A. Davis Co., Ltd. (L
A, Davis Co.) *Philadelphia, 3rd edition (197Ll)], or delayed-type hypersensitivity to tyrosine arsanilate (Niss Laskowit (8)).
, LaSkOWitgl) * J.N. 29
1 (1963) Oyohies Raskowaits, Science, 155s 350 (197
5) Reference]. Furthermore, compounds of formula 1 may be used in combination with mineral oil and vegetable oil t-L when incorporated into vaccine (or pacterin) compositions that also contain mineral oil or vegetable oil.
Animals that do not exhibit the adverse side effects normally observed when used with POA-containing vaccines (or pactelins).

Furthermore, compounds of formula 1 may themselves be used to infect infectious organisms, such as Klabsiella pneumoniae.
ias).

Escherichia coli (IC5cherichia coli),
Can1liaa albicans
* or Staphylococcus aureus
Ccus aureus).

A novel figurine of formula 1 can be produced according to the generally indicated reaction route below: In each of the above formulas, R%RI, R2X and (≦) are as defined above; R3 is 1 to 22; R4 and R5 are the same or different acyl groups having 1 to 22 carbon atoms; R6 is hydrogen or an alkyl group having 1 to 16 m carbon atoms; ;R? is hydrogen or methyl; n is an integer of 1 or 2; ph is phenyl; and
-benzyl-L-seryl, 0-benzyl-I, -)leonyl, L-methionyl, S-benzyl-L-7steinyl, L-phenylalanyl, O-benzyl-cy-tyrosyl, L-)lypto7a , e-carbobenzyloxy-L
-lysyl, δ-carbobenzyloxy-L-ornityl, gu-nitro L-fulginyl Nlm-benzyl-
L-histidyl, r-benzyl ester of L-glutamyl, L-glutaminyl% xa-7 Suharcyrno β-
Benzyl Ester % L-7 selected from the group consisting of sparaginyl, L-prolyl, or 0-benzyl-L-hydroxyprolyl.

In the case where n is an integer of 1 or 2 in the chemical compound of formula (13), the chemical compound FiY obtained in this way is each D-
It will be seen that the second compound is isoasparadine or D-isoglutamine.

When carrying out the above method, the D-amino acid of formula (1) is combined with an aralkyl alcohol such as benzyl alcohol, P-methoxybenzyl alcohol, P-nitrobenzyl alcohol, preferably benzyl alcohol, hydrogen chloride, P-) Treated in the presence of a strong acid such as luenesulfonic acid, preferably sulfuric acid, at a temperature of about 00 to about 40°C, preferably 20 to 25°C, for about 8 to about 40 hours, preferably 14 to 18 hours. , to obtain a compound of formula (2). The amino group of the 7-minoester of formula (2) thus obtained is well known in polypeptide synthesis technology and is generally converted into an acyl (e.g. formin, trifluoroacetyl, phthaloyl, etc.). : Urethane (e.g. t-butyloxycarbonyl, P-methoxybenzyloxycarbonyl, 2
-CP-piphenyl-isopropyloxycarbonyl,
innutinyloxycarbonyl, etc.); sulfenyl (
(0-nitrophenylsulfenyl, tritylsulfenyl, etc.); -1 or alkyl (triphenylmethyl, benzhydryl, etc.) @protected by blocking with a selectively removable group. More particularly, ( 2
) and a suitable reactant (e.g., t-butyl fluorocarbonate, t-butyl azidoformate, t-butyl phenylcarbonate, etc., preferably t-butyl azidoformate).
and a base (e.g. sodium hydroxide, magnesium oxide, tetramethylguanidine, triethylamine) in an aqueous-organic mixture or in an anhydrous organic solvent (e.g. dioxane, tetrahydrofuran, dimethylformamide, dimethylsulfoxide, etc., preferably anhydrous dimethylsulfoxide). etc., preferably triethylamine) for about 12 to about 48 hours, preferably 20 to 24 hours, at about 0° to about 40°C, preferably 20 to 25°C. The protected amino ester N(Z-t-butyloxycarbonyl derivative is entirely prepared.

To form an amide bond, the carboxyl group of the protected amino ester of formula (3) is reacted with one of the conventional reagents (such as dicyclohexylcarbodiimide, diisopropylcarbodiimide, or other carbodiimide) with N-hydroxysuccinimide or -Hydroxybenzotriazole; Isobutyl chloroformate or other chloroformate; 2-ethyl-5-(3-sulfophenyl)-
12-oxazole-betaine or other 12-oxacillium salts; thionyl chloride or phosphorous pentachloride; 2.
z-dibyridyl disulfide/triphenylphosphine, etc., preferably with or without additives such as isobutyl chloroformate], then activated by an anhydrous organic solvent (e.g. dioxane, tetrahydrofuran, etc., preferably with or without additives such as isobutyl chloroformate). is tetrahydrofuran]
The fish was kept inside for about 30 minutes and kept at -20° to about 4° for about 12 hours.
Treatment with anhydrous ammonia for 1 hour at 0°C, preferably from about -15° to about 25°C, yields a compound of formula (4), i.e. β-benzyl t-butyloxycarbonyl-
D-isoasparagine) (n; 1) or r-benzyl t-butyloxycarbonyl-D-inglutamine) (n=2).

In the above, t-butyloxycarbonyl is a N
The a-protecting groups are well known to those skilled in the art, and appropriate methods for the class of protecting groups used (such as tifP-
) luenesulfonic acid, methanesulfonic acid, hydrogen chloride, acetic acid mifutate, ioic acid, boron trifluoride or other intermediate strength proton donors or electron acceptors inert with an organic medium, preferably acetic acid. in a saturated solution of ethyl hydrogen chloride at about -15° to about 4 CJ°C for about 1 to about 60 minutes, preferably about 1 to about 1 at about 15° to about 25°C.
The protected isoasparagine β-benzyl ester or isoglutamine of formula (4) is treated for 5 to about 30 minutes and then diluted with a solvent having a low dielectric constant, such as hexane, ether, benzene, etc., preferably ether. removed from the r-benzyl ester compound, (5
A compound of formula j is formed, namely β-benzylisoasparaginate hydrochloride (n=1) or r-benzylisoglutaminate hydrochloride (n=2).

Next, the t-butyloxycarbonylated product of formula (6) is reacted with the compound of formula (5) to obtain compound '91J of formula (formula). This reaction is carried out in a polar solvent (ethyl acetate, acetonitrile, tetrahydrofuran, dimethyl formamide, preferably dimethylformamide] in the presence of a base (e.g., triethylamine, diisopropylethylamine, N-methylmorpholine, etc., preferably diisopropylethylamine) at from about -10° to about 80°C for about 6 u min. The reaction is carried out for about 4 hours, preferably at about 10° to about 25° C. for about 30 minutes to about 1 hour, by addition of suitable additives (such as the auxiliaries described above in connection with the preparation of compounds of formula (4)). to produce an NO-protected dipeptide with n; 1 or 2 and xl as defined above. Alternatively, t as shown in formula (6)
-Other NGs in ftU of butyloxycarbonyl group-
Protective groups such as those described above for the preparation of compounds of formula (3) can also be used.

N-acetylglycopegutide (R=R1=H, R
2=0M3. n = i or 2, X and R6
is as defined above] can be obtained as exemplified in the reaction route for producing formula Q3 from formula (8).

The raw material for this reaction route is benzyl 2-acetamido-4,6-penzylidene-2-deoxy-α- of the formula (8).
D-glucopyranoside, and the manufacture of this compound is carried out by B.H. Gross and R.W.
Loz ) by J Org Chemi (J, Org
, Chom, )t 32. 2762 (1967)
It is described in h. All of this sequential process can be carried out equally well using β-benzyl glycoside corresponding to formula 87 as a starting material; the synthesis of this starting material is also described in J.Org. Chem., 32, 2762 (1967). These steps are all carried out under conditions generally comparable to those described in the context of using the α-benzyl glycoside of formula (8) as the entire starting material.

Alkylation of the free hydroxyl group of formula (8) is carried out using a suitable base such as sodium hydride, potassium hydride, etc. in an inert organic solvent such as dimethylformamide, tetrahydrofuran, dioxane, etc. Ll'O for about 3 U minutes to about 3 hours, preferably in dimethylformamide with a width of about 10° and about 3 LI
This can be easily accomplished by first forming the anionic form by treatment at 0.degree. C. for about 15 to about 60 minutes. This anionic gold is then treated, preferably in the same solvent, with a linear a-halocarboxylic acid having 2 to 18 carbon atoms (typically chloroacetic acid, α-chloropropionic acid, α-bromopropionic acid, α-bromopropionic acid, α- -Chlorbutyric Ml, CI-proA butyric acid%
α-bromovia, α-bromocaproic acid, α-bromyristic acid and α-bromostearic acid]
react with. This step is carried out in dimethylform 7amide at about 20° to about IL10°C for about 15 minutes to 5 hours.
Advantageously, the reaction is carried out preferably at about 5 J' to about 80°C for about 0.5 hours to about 2 hours, or from about 20°C to about 30°C for about 10 hours to about 48 hours.

The resulting compound of formula (9) (R"=CH, , R)=H, R6=as defined above) is then isolated by conventional collection methods and crystallized or as known to those skilled in the art. 5 Sakaki is made by other methods.

Alternatively, the compound of formula (9) (R2=CH3,R)=
Convert aR6=@(which is the same as the above definition) into equation (9) &
'Jfttt corresponding methyl ester t R2= R
' := CH3) by the usual method, and then by the usual method,
For example, it is preferable to purify by fractional crystallization or chromatography on silicic acid, and the method for producing this methyl ester is the acid of the formula (9) (R''-=CH3,R)=H)
with diazomethane, methanol, tetrahydrofuran,
dioxane etc., preferably in tetrahydrofuran.
at a temperature of from about 20° to about 40°C, preferably from about 20° to about 25°C, for about 5 minutes to about 2 hours, preferably about 15 minutes.
The reaction is allowed to proceed for about 30 minutes, using methyl iodide or dimethyl sulfate and a solvent (e.g., tetrahydrofuran, dioxane, dimethylformamide, etc.) in the presence of a suitable base (e.g., potassium carbonate, sodium hydroxide, triethylamine, etc.). Approximately 3U at 5° to approximately 50°C
This includes a reaction in which the reaction is allowed to proceed for a minute to about 16 hours. Preferably, this esterification is carried out at about 2LI' to about 25°C, followed by sequential addition of water and dimethyl sulfate.
The reaction is carried out in situ in the alkylation medium for 4 hours to about 48 hours.

In another embodiment of this process, alkylation of the free hydroxy group of formula (8) is carried out to form the anionic form prepared as described above into a straight chain α-
Halocarboxylic acids (typical examples include methyl chloroacetate, methyl 2-bromopropionate, methyl 2-bromobutyrate, 2-
Methyl bromocaproate, methyl 2-bromocaprate, methyl 2-bromomyristate, and methyl 2-bromostearate) are reacted with esters (eg, methyl, ethyl, n-propyl, etc.). This method is particularly useful for the production of compounds of formula (9) in which R6 is a straight-chain furkyl group having 5 tanned 16 carbon atoms; It is advantageous to directly prepare the methyl ester of (9
The methyl ester of formula ) is purified by crystallization, chromatography on silicic acid, or other methods known to those skilled in the art. This reaction is carried out in an inert organic solvent such as dimethylformamide, tetrahydrofuran, dioxane, etc.
It is carried out at 100℃ for about 15 minutes to about 24 hours.
Preferably the dimethylformamide width is from about 10° to about 30°.
℃ for about 6 U minutes to 2 hours. When used, a stirred solution of the appropriate ester of a linear
A high yield of the compound of formula (9) is obtained by dropwise addition of a dimethylformamide solution of the anionic compound of formula (8).

The compound of formula (9) (R2=Rte=CH3) is then heated at about 20°C in a mixture of water and a suitable organic solvent (such as methanol, ethanol, dioxane, tetrahydrofuran, etc., preferably methanol). at the reflux temperature of the solvent mixture, preferably at a temperature of about 60° to 90°C, about 1
tnL for u minutes to about 4 hours, preferably about 15 minutes,
1 hour with a suitable base (e.g. sodium hydroxide, potassium hydroxide, barium hydroxide, etc., preferably hydroxide) IJ
(9)
Carvone@(R'l=C)i3. R? =H)
Kimf, 6° Compound of formula (9) (R2=CH3,R
) = H) and (1G various suitable protected benzyl α-aminoacyl-D-inglutaminate and benzyl α-aminoacyl-D-isoaspartate derivatives (in the formula zlH glycyl, L-alanyl, L -Baryl,
L-leucyl, L-inleucyl, 1. -a-aminobutyryl, 0-benzyl-L-seryl, 〇-benzyl-L
-) leonyl, L-methionyl, 8-benzyl-L-cysteinyl, L-phenyl-alanyl, 0-benzyl-
L-tyrosyl, L-)! j7') Phanyl, 8-carbobenzyloxy-L-lysyl, -1-carbobenzyloxy-L-ornityl, gu-nitro-L-fulginyl
Nlm-benzyl-L-histidyl, :b-/rutamyl.

β-benzyl ester of r-benzyl ester, L-glucheminyl, L-7 spartyl % L-7 sparaginyl,
L-prolyl and 0-benzyl-L-hydroxyprolyl, where n = 1 or 2) can be carried out using various techniques well known to those familiar with the art of peptide fractionation. This is done by @i.e. for example H1QI
The dipeptide hydrochloride derivative of the formula is converted to the free base form by addition of a suitable strongly basic tertiary amine as diimpropylethylamine. The resulting free aminated compound corresponding to the compound of the formula (9) is then combined with the carbonyl group of the compound of formula (9) (for example R2=CH3, R?=H and R6 is as defined above) and, for example, dicyclo A carbodiimide such as hexylcarbodiimide or diisopropylcarbodiimide is used for condensation in an inert organic solvent such as dimethylformamide, dichloromethane, ethyl acetate, dioxane, acetonitrile, etc. Optionally and preferably, this reaction ti-hydroxybenzotriazole, N-hydroxysuccinimide,
This is carried out in the presence of suitable well-known additives such as P-nitrophenol, pentachlorophenol, etc.

According to the present invention, from about 2 to about 6 L in dimethylformamide at about U° to about 50° C. for 1 hour, preferably about 10
It has been found that condensation using diisopropylcarbodiimide and 1-hydroxybenzotriazole at a temperature of 10 to 40 hours at a temperature of 60 DEG to 60 DEG C. provides excellent results. The resulting product of formula α9 (R6Xi and n are as defined above) is isolated and made t# by conventional methods including crystallization and chromatography on silicic acid. Alternatively, the product of formula tII is Free base and (9
The condensation with compounds of the formula (R?=H) can also be carried out by other methods well known in peptide technology. Thus, for example, the use of a moisture anhydride derived from a compound of formula (9) in the presence of a chloroformate (e.g. inbutyl chloroate) or Ifil-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. is effective. Also useful are other types of carbonyl activation, such as those occurring by using triphenylphosphine or oxacillium salts in the presence of dibyridyl disulfide or carbon tetrahalides. A general overview of such md&methods is provided by IaI S. Klosner and M. Bodansky (Eng., 8.1au).
Sner and M. Bodansky), Synthesis, 453 (1972)
and J.H. Jones (J.H.).

Jonesa) %Chemistry and Industry
), 723 (1974).

The reaction sequence proceeding from formula (8) to formula (9) (R?=H) can also be varied at will to allow modification of the nature of the N-acyl function, R2CQ-.

That is, the N-acetyl functional group of formula (8) is converted from J. Org. Chem., 32.
2762 (1967), hydrolysis with a strong base produces benzyl 2-amino-4,6-penzylidene-2.
-deoxy-41-D-glucopyranoside, formula (8A)
It is possible to generate . This method can also be applied to the β-anomer of formula (8), as also described in J. 2-deoxy-β-D-glucopyranoside is (aA
) can be used in subsequent reactions under essentially the same conditions as described for the reactions starting from the formula. Selective N-acylation of compounds of formula (aA) occurs by reaction with carboxylic anhydride in a lower alcohol, preferably methanol. The acid anhydrides include straight-chain and branched aliphatic carboxylic acids R2C0OH, where R2 contains 1 to 21 carbon atoms, and can optionally be methoxy, benzyloxy, halogen (e.g. fluorine), or aryl group. the aryl group having up to 10 carbon atoms and optionally substituted with lower alkyl, lower alkoxy or halogen groups]
Including those derived from. Therefore, this acyl group R2Co- is formyl (produced using R"" H1 formic anhydride-acetic anhydride), acetyl, trifluoroacetyl, methoxyacetyl, butyryl, imbutyryl, valeryl, octanoyl, lauroyl, myristoyl, valmitoyl, Behenoyl, phenylacetyl, benzoyl, P-fluorobenzoyl, P-methoxybenzoyl,
The exact conditions for this selective N-acylation, including groups such as naphthoyl, 7-medoxynaphthoyl, etc., will depend on the reactivity of the acid anhydride used. Generally, this reaction is carried out using methanol ranging from about 2 Ll' to about 1 Ll' at about 65°C.
The reaction proceeds satisfactorily in a period of from 5 minutes to about 6 hours, and the majority of the reaction can also be carried out under rapid flow for a period of from about 60 minutes to about 60 minutes. In the case of highly reactive acid anhydrides such as trifluoroacetic anhydride, the reaction is preferably carried out in an inert solvent such as tetrahydrofuran using a slight excess of the anhydride. the law of nature.

The compound of formula (8B) of the above reaction is then alkylated using sodium me-halocarboxylate or methyl a-halocarboxylate and sodium hydride in dimethylformamide as described above to give R? ==
Various benzyl 2-acyl amides-4 of formula (9) where H and R2Co- and R6 is as defined above
e 6- o-Benzylidene-3-o-(1-carboxyalkyl)-α-glucobyranoside is produced.

Coupling of these derivatives with the free base derived from the four-hole is achieved by the same method described above for the formation of compounds of formula (9) (R2=CH3), and R''CO-
A blocked glycopeptide of the α9 formula is produced in which the acyl group, XI 7 i noacyl group, n and R6 are as defined above.

Removal of protecting groups from α9-type blocked glycopeptides is accomplished by catalytic hydrolysis using a suitable palladium catalyst. This reaction is usually carried out in an acidic medium under a pressure of 1 to 2 atmospheres of hydrogen at a temperature close to room temperature using 5 to 10 m of palladium on charcoal, palladium on barium sulfate, palladium black or related catalysts. Preferably, a total of palladium on charcoal catalyst in about 60 to about 90% aqueous acetic acid is used. Under these conditions, removal of all protecting groups is complete within about 24 to about 72 hours. This removal reaction can be followed by thin layer chromatography if desired, and thus R2 of the compound of formula 04 (R=R1=1(, R6 and R''CO- are as defined above, but In the case of dimethyl in penzyl, this be/zyl ether is simultaneously cleaved to form a compound of formula 04 in which R2 is hydroxymethyl, and X is glycyl, L-alanyl, L-valyl, L-leucyl, L
-inleucyl, L-α-aminobutyryl, L-seryl, L-tononyl, L-methionyl, L-xf-niru, L-phenylalanyl, L-+rosyl, L-)liptonanil, L-lysyl, L-ornityl% L-fluinyl, L-histidyl, Ia-glutaminyl, L-glutaminyl, L-7 subartyl 5L-1 sparaginyl, L-
7'lolyl or L-hydroxyprolyl) [e.g. 2-(2-acetamido-2-deoxy-D-glucose-3-o-yl)-D
-propionyl-L-valyl-D-isoglutamine]. These compounds are manufactured by Biolex (B.
X) 7 Ll (trade name) (weakly acidic polyacrylic acid resin) by conventional chromatographic methods such as chromatography on a column. In some cases, small violet impurities cannot be completely removed by this chromatography alone. In such cases, Doweck/C (Dowe
x) All basic impurities were effectively removed through a column of 5 Ll (H”) C6 (sulfonated polystyrene crossed with divinylbenzene resin) or Amberli tθ XAD-2
All partially deblocked glycopeptides are separated by chromatography on a column containing L (polystyrene cross-linked with 2% divinylbenzene) in water or aqueous methanol.

Another aspect of the invention is a glycodipeptide of formula 0.
, 6-di-0-acyl derivative (wherein R and R1 are the same or different acyl groups, xlR'' R6 and n are as defined above).

Suitable acyl groups include straight-chain or branched alkyl, aralkyl, alkaryl or aryl groups containing 1 to 21 carbon atoms, such as acetyl, octanoyl, stearoyl, behenoyl, benzoyl and the like.

Furthermore, according to the present invention, 10,000 of R or R1 is an acyl group as defined above, the other (R1 is R1) is hydrogen, and X%R2R6 and n are as defined above. α
A selective preparative method for the 4-o- and 6-〇-acyl derivatives of formula 3 has been developed for the synthesis of all three compounds, in which protected glycopeptides of formula α9 (with the exception of zl R2R6 and n are as defined above] using selective acidic hydrolysis of the o-benzylidene functionality. These treatments include a temperature of about 50°C to about 100°C.
treatment with 50-90% acetic acid, formic acid or propionic acid, or short-term treatment with a strong acid such as about 90% difluoroacetic acid at room temperature. Advantageously, the reaction is carried out at about 100°C for about 5 to about 8 minutes using about 60 to about 80% acetic acid. Next, the usual finishing treatment is carried out, and in other respects it is sufficiently protected (12
A) The 4,6-diol of formula is isolated and purified by crystallization or chromatography on a suitable adsorbent such as silicic acid.

Acylation of both free hydroxyl groups of formula (12A) is conventionally activated such as theacyl anhydride, acyl chloride, acyl cyanide or acylimidazolide in a suitable inert organic solvent in the presence of a tertiary base. This is achieved by treatment with two or more molar equivalents of acylating agent. Suitable solvents include pyridine, methylene chloride, acetonitrile, dimethylformamide, tetrahydrofuran, dioxane, etc., and suitable bases include -Magiridine, triethylamine, N-methylmorpholine, diisopropylethylamine, and the like. It is generally advantageous to use 2 to 10 molar equivalents of either the acyl chloride or acylic anhydride in whole pyridine at room temperature for about 10 to about 24 hours, so that the resulting R4 and R6 are in the same place as R and R1. A 4,6-di-0-acyl derivative of formula (12Ef) with the same acyl group as similarly defined is isolated and purified by crystallization or chromatography on an adsorbent such as silicic acid. Removal of the protecting group from the compound of formula (12)n) uses a palladium-derived catalyst as described above for the conversion of the compound of formula α9 to the compound of formula α3 (wherein R and R1 are hydrogen). Achieved by catalytic hydrogenolysis.

The resulting product is a compound of formula 03, where R'' R6X and n are as defined above, and R and R1 are the same acyl group, except that R2 also includes hydroxymethyl.

Alternatively, a slight excess (approximately 1.0 to 1.2 molar equivalents) of acylic anhydride, acyl chloride, acyl chloride or acylimidacyline as described above!! used in active organic solvents (12A
Acylation of the compound of formula ) results in selective acylation of the primary 6-hydroxyl group.

This reaction is preferably carried out by slowly adding the acylating agent to a solution of the compound of formula (12A) at about 00 to about 2 Ll'C, and the reaction is carried out by thin layer chromatography until the starting material is substantially consumed. tracked by Next, finishing by a conventional method, formula (12F') (wherein R3 is an acyl group defined in the same way as R and R1,
n% R2 and R6 are as defined above] 6-
The whole 0-acyl derivative is isolated and chromatographed on a suitable adsorbent, such as silicic acid, until it is cleaved by crystallization. Catalytic hydrogenolysis of compounds of formula (12F') was carried out as described above using a palladium-derived catalyst, <
Formula 13 (wherein R and R2C0- are acyl groups as defined above, but benzyloxyacetyl is excluded and hydroxyacetyl and (glycolyl) groups are also added instead, R1 is hydrogen) Obtain the compound.

In order to selectively introduce an acyl group into the 4-o-position, (
12A) It is necessary to selectively attach a protecting group to the 06 hydroxy group of formula 12A). This can be conveniently accomplished by using trityl or Tj&ditrityl ether, using techniques well known in the carbohydrate chemistry field. In other words, pyridine lfc is prepared by reacting the chemical compound vlJt-) of formula (12A) with triphenylmethyl chloride (trityl chloride) or with anisyl diphenyl in an inert solvent such as dimethylformamide, dimethyl sulfoxide, acetonitrile, etc. containing pyridine. React with methyl chloride (methoxy7 trityl chloride), dianifluphenylmethyl chloride (dimethoxytrityl chloride) or trianisylmethyl chloride (trimethoxytrityl chloride) (see M. Smith et al. CM for the use of the reactants of cIL et al.).

Sm1thθt am)'s J, Amer, Chim, 8oc, J,
84 s 43 [see J (1962)].

Generally, this reaction can be carried out at about 10° to about 5 U°C, advantageously carried out in pyridine at room temperature.

The length of time for this reaction depends on the reactants used and varies from about 2 to about 5 hours for trianisylmethyl chloride, and from about 6 to about 5 hours for triphenylmethyl chloride. It changes in about 6 days. generated Cl2B)
6-o-trityl or substituted 6-o-)rityl ethers of the formula can be readily isolated by conventional techniques such as precipitation, crystallization or chromatography on a suitable adsorbent such as silicic acid. In formula (12B), R6, Xl n
and R2C0- are as defined above.

(Acylation of the free 4-hydroxyl group of formula 12B3 is carried out using conventional acylating agents such as 7-syl chlorides, acylic anhydrides, acyl cyanides or acylimidazolides derived from acids in which the acyl group R3 is as defined above. The reaction is carried out as described above for acylating a compound of formula (12A) to form a compound of formula (12B), except that the molar ratio of the acylating agent is optionally lowered, such as from about 1 to about 5 equivalents. It can be carried out under the same conditions as those previously used.

Generates the formula (12C) (in the formula R"R3R6K" O!
and n are as defined above] and cleaved by crystallization, precipitation or chromatography on a suitable absorbent such as silicic acid.

Selective hydrolysis of trityl ('! or substituted trityl) groups can occur under slightly acidic conditions. This is it
Advantageously, this is carried out by treating with 70% acetic acid at ILJ [J° C. until a clear liquid is formed and for a further approximately 2 minutes. These conditions can be varied by those skilled in the art with respect to temperature, time and acidity, except that sensitive conditions should be avoided.

When using a methoxytrityl-derived metal rod corresponding to formula (12C), the conditions for this acid hydrolysis are very mild, and the use of 80 thiacetic acid for about 5 minutes at room temperature for about 5 hours. It is enough. Generates the formula (12D) (wherein Xl
n% R' R3 and R2 are as defined above] is isolated and purified by crystallization or chromatography on silicic acid.

For further acylation of the 6-hydroxy group of compounds of formula (12D), the acyl derivatives derived from carboxylic acid R40H can be prepared using This is accomplished using a chemical agent. (12JiiJ formula (wherein the acyl groups R4 and R5 are the same or different as defined above, R2R
6% Xl and n as defined above] can be isolated and purified by conventional techniques. Alternatively, the
By further acylating the compound of formula 12FJ using a sylating agent, a compound of formula 12EfJ is produced. Again, a compound of formula 12I!
and R5 are the same or different as defined above, and R'' R6Xi and n are as defined above] is isolated and prepared by conventional means.

Removal of protecting groups from compounds of formulas (12D), (12K) and (121F) was carried out using a palladium-derived catalyst in acidic medium as described above for the conversion of compounds of formula L12A) to compounds of formula t13. This is achieved by catalytic hydrogenolysis. The product formed has the aJ formula (wherein R
6z%n and R2C0- are as defined above, Jl
R and R1 are either the same or different acyl groups, or -1 of R or R1 is hydrogen and the other is an acyl group. In both cases, A'/AIMR% R'' R6 and )L2CO- are as defined above except that a hydroxyacetyl (glycolyl) group is added in place of the benzyloxyacetyl group. These compounds of formula 0 is isolated by precipitation from methanol by addition of a less polar solvent such as ethyl acetate or ether, then (1
2A) Further purification by chromatography on Biolex 70 or Amberlite XAD-2 as described above for conversion of compounds of formula to compounds of formula Q. Obviously, other chromatographic techniques well known for peptides and glycopeptides can alternatively be used to prepare these N.

Isolation of the compounds described herein may be performed as desired by any suitable separation or preparation technique, such as extraction, crystallization, thin layer chromatography, thick layer chromatography or column chromatography, or alternatively This can be done by a combination of these methods. As specific examples of suitable separation and isolation methods, the following examples may be cited. However, other equivalent separation or isolation methods can of course also be used.

Temperature means a temperature of about 15° to about 25°C.

The invention may be further understood from the following non-limiting examples.

Example 1 Concentrated sulfur i150dt#ll; water ether 5LJOrtdv
c Carefully add and then add benzyl alcohol 50011J'lk to this mixture. The ether gold was removed by vacuum evaporation and the residue was added in portions of 679 t'' of D-7 subvalanic acid (1, n=1) over 0.5 h. The resulting suspension was stirred at 22° C. for 16 h to clarify. To this vigorously stirred solution, add 1 part of 95% ethanol.
Add j and 250d of pyridine. Crystallization begins after a few minutes and the mixture is kept at 0° C. for 20 hours. Collect the product by filtration, wash thoroughly with ether, and 1) 2°
~1) A white solid with a melting point of 7°C is obtained at 77.99°C.

The product was recrystallized from water 11 containing 10 drops of pyridine and had a melting point of 220° to 221°C and [α]fi” −2
β-benzyl D-asparte) with 8,1° (101119/HLt, IN hydrochloric acid) (2, n=1)
44.31t-obtained.

In a similar manner, using stoichiometric equivalents of D-glutamic acid (1, n=23) instead of D-aspartic acid, 1
Melting point from 61 ° to 162 ° C and Cab D - 19,5 ° (
r-benzyl D-glutamic acid) with 6#/d acetic acid)
(2, n=2) 56.3 g is obtained.

Example 2 Anhydrous dimethyl sulfoxide 25051) in a 5L101j round bottom flask. β-Benzyl D-7 Spartate (2,
n=1) 11.15#,) ethylamine j3.5m
and t-butyl azide) 91) Hold the resulting solution at 22° C. for 42 hours. The reaction mixture was then poured into 700 au of water and the mixture was dissolved in ether 2LIO.
Extract 3 times with JLJ. The aqueous phase is cooled with ice, 200 JLlt of ethyl acetate is added, then the pH of this aqueous phase is
Addition of citric acid brings it to 2.8. The phases are separated and the aqueous phase is further extracted three times with 2υ[1jlj of ethyl acetate.

The combined organic phases are extracted 5 times with 2[J[ld] water and successively with 2[j[Jlj] saturated brine and dried over magnesium sulfate. The ethyl acetate solution was filtered, the filtrate was evaporated to dryness, and the solid residue obtained was crystallized from ethyl acetate-hexane, with a melting point of 102° to IL 13°C and [
β-benzyl t-butyloxycarbonyk-1)-asparte) with α]r19.4 (1419/', dimethylformamide) (5, n=1) 16
-29 is obtained.

Elemental analysis: For CzaHzxNOa (323,34), calculated values: C, 59,43; H, 6,55; N,
4.33 Actual value: c, 59.51; a, 6.39; N
, 4.28° In a similar manner using a stoichiometric equivalent of r-benzyl D-glutamic acid (2, n = 2) instead of β-benzyl D-aspartate, r −
Benzyl t-butyloxycarbonyl-D-glutamate (3jn-2) 16.9! Get lft.

This product has sufficient purity to be used directly in the next step.

Example 3 β-benzyl t-butyloxycarbonyl-D-aspartate (3, n=1
) 3-2311 and triethylamine 1, (5au
The magnetically stirred solution was cooled to -15°C. A solution of inbutyl chloroformate 1.55+j in tetrahydro7ran 5a was added dropwise to this solution, and the temperature was increased to ft2.
Maintain at -15°C for 5 minutes. Dry ammonia gas t' is bubbled through the reaction mixture while maintaining the temperature at -15°C for 3 J minutes. The temperature of the reaction mixture is then increased to 0° C. over a period of 15 minutes, at which point the flow of ammonia gas is stopped. Evaporate tetrahydrofuran under reduced pressure,
The residue is partitioned between water (8011) and ethyl acetate (200d). Ethyl acetate phase t-51 is washed successively twice with 50M aqueous sodium bicarbonate solution and then twice with saturated brine 501111, then dried over magnesium sulfate.

The filtered solution was evaporated to dryness and the resulting solid 3.22.9
', recrystallized from ethyl acetate 501),
Melting point of 6LI'O and [α]D +1). LI'(
I LJII91jLl, dimethylformamide) with benzyl t-butyloxycarbonyl-D-in aspara Q'un-) L4, n=1) 2-72
jlk get original search analysis 2C engineering aHgzN20s (52
2,35), the calculated value 2C,59,61;H,6
,88;N,8.69I measured value 2c,59.8i ;)I
, 6.8LI; N, 8.87 In the same manner, instead of β-benzyl t-butyloxycarbonyl-D-7 sppartate, Chemodoron's Or-benzyl t-butyloxycarbonyl-D-gludame -) (3, n=2), melting point of 125° to 126°C; [α) D-2,69
Benzyl t-butyloxycarbonyl-D-4 soglutaminate (4, n=2) with (13,3 m9/at, dimethylformamide) 1-92flt- is obtained.

Example 4 Add 2.95 g of A, benzyl t-butyloxycarbonyl-D-inglutamine (4, n=2) to 14 g of a saturated solution of hydrogen chloride in ethyl acetate. The reaction mixture was heated to 22°C.
The product was precipitated by addition of ether 80°C, the precipitate was collected by centrifugation, carefully dried under vacuum, and r-pencyl D-isoglutaminate hydrochloride (5゜n=2)2・, 66 g was obtained. This product is used in the next step B without further purification.

B, dry dimethylformamide)'L dissolved in Qm
-Butyloxyluponyl-L-alanine (6°X1=
1.72 g of 1-hydroxybenzotriazole and 2.19 g of dicyclohexylcarzadiimide are added to 2.01 g of L-alanyl). The resulting mixture is 22
℃ for 1.5 hours and then filtered to give γ-benzyl D-
Inglutaminate hydrochloride (5゜n=2) 2.56'j
and diimpropylethylamine 1.55 m/dimethylformamide iQm/@ solution. The insoluble cake obtained from this filtration was treated with 5 t of dimethylformamide.
Add the washing solution to the reaction mixture and keep at 22° C. for 1 hour.

The reaction mixture is poured into 500 g of water and the resulting solution is extracted twice with 250 g of ethyl acetate. Pour the collected organic phase into 2 parts of water.
00-, 6 times with 150 ml of 5% aqueous potassium bicarbonate solution and 150 ml of saturated saline solution successively.

The ethyl acetate solution was then dried over sodium sulfate, filtered and evaporated to dryness to give a solid product, which was crystallized from a mixture of ethanol and ether at 140°
Melting point of ~141 °C and [α] 220 ° (7,6 Da/
benzyl L- with wl, dimethylformamide)
Detyloxycarbonyl-L-alanyl-D-inglutaminate (7, n=2t X”=L-alanyl)2
．． Obtain 2g.

Elemental analysis: C2oH26N306 (407,45)
For, the calculated value: c, 58.95; H, 7,
1); N, 10.31 Actual value: c, 59.1
1; H, 6,80; N, 10.50 In the same way, butyloxycarbonyl-L-alanine is replaced by a stoichiometric equivalent of b-butyloxyluponyl-L-valine, -butyl oxycarbonyl-L-α-7minobutyric acid, t,-tetyloxycarbonyl-O-benzyl-L-)
leonine, 11, --If-ruoxycarbinyl-L-phenylalanine, t-butyloxycarbonyl-〇-benzyl-L-tyrosine, -butyloxycarbonyl-ξ-carbobenzyloxy-L-lysine, t-butyloxycarbonyl-l-benzyl-L-lysine, L-proline and L-butyloxycarbonyl-〇-benzyl-L-hyr
oxyzololine to obtain the corresponding protected dipeptide derivative: penzyl L-butyloxycarbonyl-L-valyl-D
- Inglutaminate, melting point: 152°-156°O;
Cα]” +9.1' (10Tn9/m/, dimethylformamide): Elemental analysis: Calculated value for C2□H33N306 (465-50): c, 60.67; H, 7,6
4; N, 9.65 Actual value: c, 60.88;
H, 7,90; N, 9.67 to 7 sil F--butyloxycarbonyl-L-α-aminobutyryl-D-
Inglutaminate, melting point: 102° to 104°C: [
α]”+ 6.1° (10 Da/mA', dimethylformamide); Elemental analysis: C21H31N306 (421-50)
Calculated value for: c, 59.84 ; )(,7,
41; N, 9.97 Actual value: c, 59.60
H, 7,56; N, 10.00 Pencil-butyloxycarbonyl-benzyl-L-)leonyl-D-isoglutaminate, melting point = 11)° to 120'
O; Cα125+7.1° (5,0ff9/It/, dimethylformaminr): Elemental analysis: C28H3FN3
0ft 527.65), calculated value: c,
65.74; H, 7,07; N, 7.96
Actual value: c, 66.59; He 6.81
; N * 7.84 Benzyl-butyloxycarbonyl-L-phenylalanyl-D-inglutaminate, melting point 169-140°C; [α]',' -10,7
°(10m97-, dimethylformamide): Elemental analysis: C26H33N306 (,483,57
): Calculated value for c, 64.58; )(,6
, 88; N, 8.69 Actual feed value: C, 64, 62;
[
α]24+4.2° (4,6rn9/ml s dimethylformamide): Elemental analysis: C33Hss+N50
Calculated value for 7 (5B9-66): c, 67
．． 21; H, 6, 67; N, 7.16 actual value:
C, 67,44; H, 6,88; N, 7.06 benzyl-monodetyloxycarbonyl-ξ-carbobenzyloxy-L-lysyl-D-isoglutamate, melting point 94-97°C; [ α ] 25-6.5° (311197
+aj, dimethylformamide); Elemental analysis: (,5
lHa2Naos (598,68) Calculated values for K: c, 62.19; H, 7,07; N, 9.5
6 Actual value: c, 62.19; H, 7,05;
N, 9.44benzyl L-butyloxycarbonyl-
L-prolyl-D-inglutaminate, [α)24-
Colorless oil with 9.0°D (14,4~/lJ, dimethylformamide): Elemental analysis: C22H31N306 (455,49)
Calculated value for K: c, 60.95; H, 7,2
1; N, 9.69 Actual value: c, 61.06; H
, 7,49; N, 9.85 and benzylmobutyloxycarbonyl-〇-benzyl-L-hydroxyprolyl-D-inglutaminate, [α)''-2,4'
L 5 da/-, dimethylformamide tr) 6 oil.

Example 5 A, benzyl 2-amino-4,6-0-benzylidene-
2-deoxy-α-glucopyranoside L8A) 5.57
1 and benzoic anhydride 2.499 methanol 200-
Heat the solution under reflux for 45 minutes. The resulting mixture is 2
Cooled to 2°C, the crystalline solid was collected by filtration, washed with methanol, and then recrystallized from hot methanol to give benzyl 2-benzamide-4°6-0-benzylidene-2-deoxy-α-D-glucobyranoside ( 8B, a”=ph)3.
Obtain 46 g, melting point 218°-220°C: [α”l'
,'+119° (10■/d%t lysine); Elemental analysis: C2FH2F No., H2O (479,54
): c, 67.66; a, 6.1
0; N, 2.92 Actual value: c, 68.09
; H, 5, EH; N, 2.87B, benzyl 2
-amino-4,6-0-penzolidene-2-deoxy-
A tetrahydrofuran 1001Lt/I solution containing 6.571 α-D-glucopyranoside (8A) and 2.61 g of trifluoroacetic anhydride is heated under reflux for 1 hour. The resulting solution is allowed to cool to about room temperature and then added to 1 J of ice water. The precipitated solid was collected by filtration, thoroughly washed with water, and dried under reduced pressure to give benzyl-4゜6-o-benzylidene-2-deoxy-2.
-trifluoroacetamide-α-D-glucopyranoside 4.5111 is obtained, melting point 242-246°C; [α]
25+ 115.2” (91qI/I11.dimethylformamide): Elemental analysis: C2□a2.No, p3・
Calculated value for H*O(471,44): c, 56
．． 05; H, 5, 16; N, 2.97 Actual value:
co 56.46; H, 5, 18; N, 6.
07C, benzyl 2-amino-4,6-0-penzolidene-2-deoxy-α-D-glucopyranoside (8A)
A solution of 5.4 g and 2.7 g of methoxyacetic anhydride in 25 Qml of methanol is heated to reflux for 60 minutes.

The resulting solution was then concentrated under reduced pressure and the residue
Mix with 5oat, filter and add benzyl 4.6＼-
0-Benzylidene-2-deoxy-2-methoxyacetamide-α-D-glucopyranoside (8B, R2=
-CH20CH3) 5-99 II, melting point 19
8° to 199°C; [α] 25+ 82.0° (10 calls/
1! Lt, chloroform); Elemental analysis: Calculated values for Cz3Hz7NOy (429-47): c, 64.52; H, 6,54;
N, 5.26 Actual value: C, 64,19; H, 6,
28; N, 5.06 Using step B or C in a similar manner using a suitable acylating agent in stoichiometric amounts in place of benzoic anhydride and modified according to the physical properties of the product. to obtain the corresponding 2-acylaminoglucose derivative of the following formula (8B): Benzol 2-form 7 mil'-4+6-o-benzylidene-2-deoxy-α-D-glucopyranoside penzyl 2-propionamide -4,6-0-Benzylidene-2-deoxy-α-D-glucopyranosidebenzyl 2-butyramide-4,6-0-benzylidene-2-deoxy-α-D-glucopyranosidebenzyl 2-valeramide-4,6-0 -Penzolidene-2-deoxy-α-D-gelcopi2nosidebenzyl 2-cafuramido-4,6-Q-benzylidene-2-deoxy-α-D-glucopyranoside, melting point 20
2° to 204°C: [α] 23-1-108.2° (I
Qlllil/d, dimethylformamide): Elemental analysis: C26H33NO6・0.5 H2O (46
4,57) Kj2tfl calculated value: c, 76.22
; H, 7,58; N, 5.02 Actual value: C
, 67,54; H, 7,66; 4,6-0-Benzylidene-2-deoxy-α-D-glucogyranosidebenzyl 2-nonanamide-4,6-0-penzolidene-2-deoxy-α-D-glucopyranosidebenzyl 2-cabramide-4,6- 0-Pensilibin-
2-deoxy-α-D-glucopyranoside pe/zyl 2-lauramide 4.6-0-penzolidene-
2-deoxy-α-D-glucopyranoside penzyl 2-myristamide 4.6-0-benzylidene-2-deoxy-α-D-glucopyranoside r, melting point 18
2°～184℃: [α)”+55.5°(5m9/+a
/, chloroform): Elemental analysis: C34H4GN06・0-5H2 (857
Calculated value for 6,78): c, 70.80;
H, 8,74; N, 2.43 Actual value: c, 70
．． 50; H, 8, 61; N, 2-56 benzyl 2
-Palmitamide 4.6-0-Benzylidene-2-deoxy-α-D-glucopyranosidepenzyl 2-stearamide-4,6-〇-penzolidene-2-deoxy-α-D-glucopyranosidepenzyl 2-arachidamide 4.6-0 -Benzylidene-2-deoxy-α-D-glucopyranosidebenzyl 2-behenamide-4,6-0-benzylidene-2-deoxy-α-D-glucopyranosidebenzyl 2-benzyloxyacetamide-4゜6-0
-Benzylidene-2-deoxy-α-D-glucopyranoside, melting point 168° to 169°C; [α)25+ 72
．． 1'(5#/G,chloroform); Elemental analysis:
C2? H! 1IINO? Calculated values for (505-55): c, 68.89; H, 6,18; N
, 2.77 Actual value: C168,82: R16,06
:N, 2.90 and benzyl 2-phenylacetamido-4,6-0-penzolidene-2-deoxy-α-D-glucopyranoside.

Example 6 Benzyl 2-7cetamido 4.6-0-benzylidene-2-deoxy-α-D-glucopyranoside (8) 4
1.7 g dry dimethylformamyrs o o m
Add the e-bath solution sodium hydride in portions to 19.2 g of a 50% suspension in oil. The resulting mixture is 22
Stir at 60°C for 60 minutes, then add sodium chloroacetate 26.
59 is added and the reaction mixture is heated at 75° C. for 1 hour. The reaction mixture was cooled in an ice-water bath and diluted with ethanol 125-
is added dropwise until the bubbling subsides (approximately 30 minutes) and the resulting solution is evaporated to dryness under reduced pressure. The residue thus obtained is stirred mechanically with a mixture of 500 d of chloroform and so mt of water until the sodium salt of the product crystallizes out. The crystals were collected by filtration and resuspended in 6J of chloroform, then about 450 ml of DoveX 50 (T(+) (sulfonated polystyrene resin cross-linked with divinylbenzene) was added and the mixture was stirred for 1 hour at 22°C. The resin was filtered off and the chloroform filtrate was dried over magnesium sulfate, refiltered and then evaporated to dryness under reduced pressure to give benzyl 2-1cetamido-4,6-0-penzolidene-2-deoxy-6- o-carboxymethyl-α
-D-glucopyranoside (9, R2=CH3) 59
g, melting point 215u-216°C.

In a similar manner, benzyl 2-acetamide-4,6-
In place of 0-benzylidene-2-deoxy-α-D-glucopyranoside, stoichiometric equivalents of the various compounds obtained in Example 5 are used to obtain the corresponding compounds of formula (9).
Representative examples are as follows: Benzyl 2-benzyloxyacetamide-416-0-benzylidene-2-deoxy6-〇-carboxymethyl-α-D-glucopyranoside, melting point 1) 2° to 1) 6'C ; (α]25 +81
．． 4° (5m9/ml, chloroform): Elemental analysis: Calculated value for C3□H,,5NO, (577,61): c, 66.06; R15,90;
N, 2.49 Actual value 2c, 66.1); H
, 5,96; N, 2.58 benzyl 2-methoxyacetami)'-4,6-O-benzylidene-2-deoxy-6-0-carboxymethyl-α-D-glucopyranoside, melting point 209-210°C; [α]” + 88.
1° (10 da/d, methanol): Elemental analysis: Calculated values for C25H29NO9 (487-52): C, 61,59; H, 6,00; N
, 2.87 Actual value: C, 61,40; H, 6,40
; N, 2.77penzyl 2-benzamide-4,6-
Q-benzylidene-2-deoxy-5-o-carboxymethyl-α-D-glucopyranoside, melting point 257°~2
58°C: [α)'5+ 121.8° (10 strokes/-,
Dimethylformamide); Elemental analysis: Calculated value for C29H29N○5 (519-56): c, 67.04; H, 5,66
; N, 2.70 Actual value: c, 67.26; H,
5,56; N, 2.76 penzyl 2-trifluoroacetamide-4゜6-0-benzylidene-2-deoxy-6-0-caloxymethyl-α-D-glucopyranoside pencyl 2-caproamide-4,6- 0-Benzylidene-2-deoxy-5-o-carboxyme fk-α-D
- Glucopyranoside, m point 21) ~ 218°C; [α]”
+ 106.1° (51#/mj, chloroform); Elemental analysis: Calculated value for C28H35NO8 (515,60): C,65,48; H,6,87; N
, 2.7 + actual measurement 1st shift: Cl65.0 (S; H,
6,74; N, 2.75 and pencil 2-myristamide-4,6-o-benzylidene-2-deoxy-6-〇-carboxymethyto-α-〇-glucopyranoside, melting point 188-190°C.

Example 6' Benzyl 2-acetamido-4,6-O-benzylidene-2-deoxy-α-D-glucopyranoside (8) 4
Add 9.6 g of 100% sodium hydride (crystals) to a solution of 7 g of C in dry dimethylformamide 5QQmj.
The resulting mixture was stirred at 22° C. for 60 minutes to produce the sodium derivative of 8). On the other hand, 100%
) IJium hydride 7.0. ! i'eD, L-2-
Chlorpropionic acid (or stoichiometric equivalent, L-2
26 g of -bromozologionic acid) are carefully added to the water-cooled solution in 200 d of dry dimethylformamide.

The resulting mixture is added to the above sodium derivative and the reaction mixture is stirred at 75° C. for 6 hours. The reaction mixture is cooled to about 5° C. in a water bath and 100 d of water are added followed by 100 d of dimethyl sulfate over a period of 2 hours. The temperature of the reaction mixture is brought to 22° C. and the mixture is then stirred at 22° C. for 16 hours and poured into a large excess of water. The precipitate formed is filtered off, washed with water, then dissolved in chloroform 500- and the aqueous phase is removed. The chloroform solution was dried over magnesium sulfate and then concentrated under reduced pressure to give the crude ester (9@
R''=R6=R)=CH3) 49.6 g are obtained.

This mixture was chromatographed on a 1:9 column of prepared silicic acid and 1:1 of ethyl acetate and chloroform.
Elute with a mixture. Collect appropriate fractions, evaporate to dryness,
Recrystallization from methanol gave benzyl 2-acetamido-4,6-0-benzylidene-2-deoxy-6-0
-CD-1-carbomethoxyethyl)-α-D-glucopyranoside (9°R”=R’=R)=CH3) 26-
61, melting point 212-214°C; ((11”+
126.8° (1[1rn9/mJ in lychloroform).

Benzyl 2-acetamido-4,6-0-benzylidene-2-deoxy-6-0-CD-1-carbomethoxyethyl)-α-D-glucopyranoside (9# R2=R'
=R)=CH3) To a suspension of 26-6 F/ in 1 part of methanol was added a solution of 1 part of water, 20 I of +3 um was added, and the mixture was heated at 100° C. for 1.5 hours, Allow to cool to room temperature at this point. The resulting solution is approximately 500-
The solution is then treated with 6N hydrochloric acid until -2.5. The white solid produced is collected by filtration,
Washed thoroughly with water, dried under vacuum over phosphorus pentoxide, and then crystallized from methanol, benzyl 2-acetamide-4
,6-〇-benzylidene-2-deoxy-6-〇-(D
-1-Carphokidiethyl)-α-D-glucopyranoside (9, R2=R'=CH3, R7=H) 21.9I is obtained, melting point 269°-241°C In the same + direction, D, L- In place of 2-chloropropionic acid, a large amount of another D, L-2-noroalkanoic acid (having 4 to 6 carbon atoms) is used,
(9) Formula (R6 is alkyl having 2 to 4 carbon atoms, giving a compound with R'=l, representative examples of which are shown below: Penzyl 2-acetamido-4,6-0- Benzylidene-2-deoxy-,5-O-(D-1-carboxy-
1-propyl)-α-D-glucopyranoside, melting point 21
6°~21)℃: [α]” +126.7°t5T19
/ml, dimethylformamide): Elemental analysis: Cwe
Calculated value for HzlkJDe (485,55):
c, 64.62; H, 6,44; N, 2.8
8 Actual measurement value: c, 64.56; H, 6,22;
N, 2.98 and be/zyl 2-acetamide 4.6-0-benzylidene-2-deoxy-3-0-(D-1-carboxy-1-
(pentyl)-α-D-glucopyranosi V, melting point 262°
~266℃; [α)”5+115.2°t5W/111
4. dimethylformamide); elemental analysis: CzaHs
Calculated value for sNOa (516,60): c*
65.48; H, 6,87; N, 2.76 Actual value: c, 65.68; )(, 7,02; N,
2.77 Similarly, benzyl 2-acetamide-4,6
-0-Benzylidene-2-deoxy-α-p-glucogyranosy V is replaced by stoichiometric equivalents of the various compounds obtained in Example 5 to obtain a compound of formula (9). The representative side is shown below: Pen 2-benzyloxyacetamide-4゜<5-
o-benzylidene-2-deoxy-6-〇-(D-1-
Carboxyethyl)-α-D-/Rukogiranoshi r1 Warp point 220-221°C; [α]25+76.8°(5W9
/d, chloroform): Elemental analysis: C32H35NO
Calculated value for 9(577,61): c, 66.5
4; H, 6, 11; N, 2.45 Actual value: c
, 66.48; H*6.57; N, 2.46 benzyl 2-methoxyacetamide-4,6-〇-benzylidene-2-deoxy-6-o-(D-1-carboxyethyl)-α-D- Glucopyranoside, melting point 209-2
10°C; [α)''+107° (t!v/lj, methanol); Elemental analysis' Calculated values for C26H31NO9 (501,55): c, 62.27; H, 6,25;
N, 2.79 Actual value: C, 6,5,00; H, 6,
24; N, 2.78 benzyl 2-benzamide-4
, 6-0-benzylidene-2-deoxy-6-o-(D
-1-carboxyethyl)-α-D-glucopyranosyl
1 Melting point 275° to 276°C: [α]25+ 114.7
°(5119/ R1, dimethylformamide); Elemental analysis: Calculated values for C, soH*xNOa (555,59): c, 67.55; H, 5,86;
N, 2.65 Actual value: C, 67,56; H, 5,9
4; N, 2.54bensyl 2-trifluoroacetamide-4゜6-0-benzylidene-2-deoxy-6
-o-(D-1-carphokidiethyl)-α-D-/rucobilanoside penzyl 2-caproamido-4,6-0-benzylidene-2-deoxy-3-0-CD-1-carboxyethyl )-α-D-glucopyranoside, 5lip/d, dimethylformamide): Elemental analysis: C29H3 Non
Calculated value for (527-62): c, 66.0
2; H, 7-07; N, 2.65 actual value:
c, 66.0t; H, 6,88; N, 2.69 and benzyl 2-myrimethamide-4,6-0-benzylidene-2-deoxy-5-0-LD-1-carboxyethyl)-α-D -glucopyranoside, /1Ll, dimethylformamide): Elemental analysis: Calculated for C37H53NO8 (669,84): C,69,46; H,8,35; N
, 2.19 Actual value: c, 69.68; H, 8
, 21; N, 2.16 Examples 6″ Pencil 2-7 Cetamide 4. 6-0-Benzylidene-2-deoxy-α-D-glucopyranoside (8) 4
．． 1) Dissolve g in 5 Q ml of dry pyridine,
The DFi liquid is evaporated to dryness under reduced pressure. This process was repeated and the residue was then dissolved in 75 w1 of dry dimethylformamide and 10 (111 sodium hydride (crystals) 0.7
Add 2.91ft one part at a time and stir the resulting mixture at 40°C for 1 hour to obtain (8) IJum conductor.

This solution is then prepared from an ethanolic solution containing methyl D, L-2-promodecanoate LD, 5.0 g of L-2-bromodecanonic acid and 61 mmol of diazomethane)5.
．． Add 6 g #) dropwise over 15 minutes to a vigorously stirred solution in dimethylformamide 25+11). The resulting mixture is stirred at 22° C. for 16 hours and then concentrated to dryness under reduced pressure. The resulting residue was then treated with ethyl acetate 5
Partitioned between 00 and 250 of water, the ethyl acetate phase was further washed with 100 of water, dried over magnesium sulfate and then concentrated under reduced pressure to yield the crude ester (9°R2=R)=M
8, R'=n-c8H1)) 7.7 g is obtained.

The mixture was chromatographed on a 0400 g silicic acid column prepared in ethyl acetate:chloroform (1:9), 2. ,g
Extract with a linear solution of 2 J of ethyl acetate:chloroform (6:7). Appropriate fractions were collected and evaporated to dryness to give benzyl 2-acetamido-4,6-0-benzylidene-2-deoxy-5-010-1-carbomethoxy-1-nonyl)-α-D-glucopyranoside (9° R"=R)=M6 e R'=n-CaH1)) 2.
Obtain 461: melting point 146°-148°C; (α)”5
+89.0° (5 da/1, chloroform); Elemental analysis: Calculated values for Cs3HtsNO8 (586,75): C,67,90; H,7,77; N
, 2.40 actual value: C, 68,29; H, 7,7
7; N, 2.44 benzyl 2-7 cetamide 4.6
-0-Benzylidene-2-deoxy-3-0-CD-1
-carbomethoxy-1-nonyl)-α-D-glucopyranoside (9+ R2=R'"'Me*R'=n
-CBH77) 1.1) To a warm (approximately 60°C) methanol 65-solution of I, add a solution of 0.8 g of IJum in 55 wL of water dropwise at such a rate that no precipitate of the raw material is seen. Add. The resulting solution t- was heated to reflux for 25 minutes, cooled to 22°C, and then diluted with 6N
Adjust to 2.5 with hydrochloric acid. Approximately 2 parts ice/water to the resulting mixture
50- was added immediately and the precipitated white solid was collected by filtration, washed thoroughly with water and then dried under vacuum over phosphorus pentoxide to give benzyl 2-aceto7mide 4.6-Q-benzylidene-2-
Deoxy-6-. -(D-1-carboxy-1-nonyl)-α-D-glucopyranoside (9r R””Me
*R6=rl-C6H13@R'=H) 1.
Obtain 15 g: melting point 194°-196°C; [α)”
+94.7° (5Tn9/d, dimethylformamide): Elemental analysis: C3zHasNOa (569-70
) VC vs. Sur calculated value: C, 67, 47; Hl 7
．． 61; N, 2.46 Actual value: c, 67.5
9; H,7,62; Nl 2.55 Similarly, another methyl D in a stoichiometric equivalent in place of methyl D, L-2-promodecanoate.

Using L-2-bromoalkanoate (containing 5 to 18 carbon atoms), formula (9) (R' is alkyl having 5 to 18 carbon atoms, R'=H
), a representative example thereof is shown below: Hensyl 2-7cetamido 4,6-0-benzylidene-2-deoxy-6-0-(D-1-carboxy-1
tridecyl)-α-D-glucogyranoside: melting point 157
° ~ 159 °C: [α)” + 77.2 ° (519/d,
dimethylformamide) and benzyl 2-acetamide-4,6-0-benzylidene-2-deoxy-6-o
-CD-1-carboxy-1-heptadecyl)-α-D
-Glucopyranoside; melting point 155-158°C; [α]
25+81.4° (59/mJ, dimethylformamide).

Similarly, in place of pencil 2-1 cetamido-4,6-〇-benzylidene-2-deoxy-α-D-glucopyranoside, various compounds obtained in Example 5 of the stoichiometry were used and the corresponding ( 9) A compound of formula is obtained.

Example 7 A, benzyl L-butyloxycarbonyl-L-valyl-D-isoglutaminate (7, n=2, at 22°C. 1 ether 40 after 20 minutes
- was added, the precipitated salt was collected by centrifugation, and carefully dried under reduced pressure.
get.

B, diisopropylethylamine Q, 1) mj was added to a solution of the above benzyl L-valyl D-inglutaminate hydrochloride (10an = 2h X' = L-valyl) in dimethylformamide, and then 0.159 g of diisopropylcarbodiimide, 0.1) 8 g of 1-hydroxybenzotriazole and benzyl 2-acetamide-
4,6-0-benzylidene-2-deoxy-5-0-L
A 5 m solution of D-1-carboxyethyl)-α-D-glucopyranoside (9, R2=R6=CH,, R)=1 () in dimethylformamide) is added. The resulting reaction mixture was magnetically stirred at 22°C and the reaction was evaluated using the Kaia test.
er Leak) and add reactants until the reaction is complete. Upon completion of the reaction, the reaction mixture was poured into excess water, the solid formed was collected by filtration, washed thoroughly with water,
It was then washed twice with ethanol, dried under vacuum over phosphorus pentoxide, and benzyl 2-(benzyl 2-acetamide-4,6
-0-benzylidene-2-deoxy-α-D-glucopyranosido-6-0-yl)-D-propionyl-L-valyl-D-inglutaminate (11,n; 2.x=L
- burr λ, R2=R6=CH3) obtains 0.749,
Melting point 275-277℃ (decomposition); (α)'i5+ 76
．． 0° (I QII9/d, in methanol:chloroform, 15:85): Elemental analysis: C4xHsoN40x
Calculated value for 1 (794-84): C,61,55
; H, 6,50; N, 7.26 Actual value: C, 66
, 45; H, 6, 51; N, 7.06 Example 4 of stoichiometry for benzyl L-decyloxycarbonyl-L-valyl-D-isoglutamate in a similar manner.
to obtain the corresponding protected glycopeptyl r derivatives: penzyl 2-(benzyl 2-7cetamido 4°6-o
-Benzylidene-2-deoxy-α-D-glucopyranoside 5-0-(L)-D-7'Ropionyl-L-α-
Aminodecyl-D-isoglu: l mi$-), melting point 241-262°c; t-a]a5+95.4°(
10~/1, dimethylformamide): Elemental analysis: C
Calculated values for 41H5ON4011 (774-88): c, 65.55; He 6.50; N,
7.26 Actual value: c, 65.66; H, 6
,48; N, 7.25benzyl 2-(benzyl 2-
Acetamide-4゜6-0-benzylidene-2-deoxy-α-ri-glucobyranosidol-6-〇-yl)-D-
7'Robionyl-0-benzyl-L-tononyl-D-
Isoglutaminate, melting point 220° to 225°C: [α]
” + 76.6° (6r!I9/r/, dimethylformamide); Elemental analysis: C45HsaNaO12 (881-01)
Calculated value for: c, 65.44; H, 6,4
1; H, 6, 36 actual value: c, 65.78;
H,6,82; H,6,68benzyl 2-(benzyl 2-acetamido-4゜6-0-benzylidene-2-
deoxy-alpha-glucobylanoside 6-0-'r-D-7'ofonyl-L-puenylalanyl-D-
Isoglutaminate, melting point 276°~277'O; (α
)” +98.9 (4,5η/tnl in dimethylformamide); Elemental analysis: C46H52N4011
Calculated value for (1356,95): c, 66
．． 01; H, 6, 26; N, 6.69 Actual value:
c, 66.29; H, 6,27; N, 6.49
Benzyl 2-(benzyl 2-acetamiv-4゜6-〇-benzylidene-2-deoxy-α-D-glucopyranoside-6-o・i/L-) -D-propionyl-〇-
Benzyl-L-tyrosyl-D-isoglutaminate, melting point 276°C (decomposition); [α]24+ 85.4° (2
,59/rrLl, dimethylformamide): Elemental analysis: Calculated values for C3sHaaNtO12 (945,08): C,67,50; H,6,20; N,
5.94 Actual value: c, 67.49; H, 6,
16; N, 5.77 Penzyl 2-(benzyl 2-acetamido-4゜6-〇-benzylidene-2-deoxy-α-D-glucobyranosid-3-〇-yl)-D-propionyl-ξ-carbobenzyloxy-L- Risil-
D-isoglutaminate, melting point 256-259°C (min M
); (a]25+79.2 (10rn9/mj,
Dimethylformamide): Elemental analysis: C5tHaINsO1s (952-Q
8) Calculated value for K: c, 64.54; H,
6,46; N, 7.66 Actual value: c, 64.07
; H, 6,72; N, 7.31 benzyl 2-(pencyl 2-7cetamido 4゜6-o-benzylidene-2
-Deoxy-α-D-glucopyranoside 6-0-(L)-D-7' Robionyl-L-Zorolyl-D-isoglutaminate, melting point 1) 9° to 180°C; [α)'', 7
+ 98.2' (5W1g/ts1% zomethylformamide): Elemental analysis" C42H5ON401, (
Calculated value for 786-89): c, 64.11
; H, 6,40; N, 7.12 Actual value: c
, 64.56; H, 6,29; N, 7.01 benzyl 2-(benzyl 2-acetamide-4゜6-〇-
benzylidene-2-deoxy-α-D-glucopyranosyl P-3-0-yl)-D-propionyl-〇-benzyl-L-hydroxyprolyl-D-isoglutamate,
Melting point 75-79°C; ((1)”+93.5°(10
m9/rql, dimethylformamide) ; Elemental analysis: C49H56N4012 (8'?5-0
Calculated values for 2): c, 65.90; H, 6,
52; N, 6.27 Actual value: Ce65.71;
H, 6,55; N, 6.1) Similarly, benzyl 2-7cetamido 4.6-0-benzilysine-2-
−? "oxy-6-〇-LD-1-carboxyethyl)
-Instead of α-D-glucopyranoside, use the self-congratulatory pencil 2-7 cetamide 4.6-0-benzylizo/
-2-deoxy-6-0-cal dgoxymethyl-α-p
-/A/ Copyranoside (9, R2=CH!S)
By reacting this with each of the appropriate compounds obtained in 91J 4, the following corresponding protected glycopeptide derivatives are obtained: Benzyl (benzyl 2-acetami I''-4,6 −〇−
Benzylidene-2-deoxy-α-D-glucopyranoside-6-〇-yl)-a7,000-4-valyl D-inglutaminate, melting point 257-239°C; (α)”
+80'9° (,9m9/ml, dimethylformamide): Elemental analysis: C42H32N4011 (774-84
) calculation for ([: Ce65.55 ; H,6,
50; N, 7.25 Actual value: c, 65.45;
H, 6,51; N, 7.06 pen 2 (benzyl 2-
Acetamide-4,6-〇-benzylidene-2-deoxy-α-D-glucohyranosido-6-〇-yl)-acetyl-L-α-aminodecyl-D-isoglutaminate, melting point 218-260°C: [α]” +81.8
° (10mg7rnt, dimethylformamide); elemental analysis 204°I (calculated values for 48N4011 (760,86): c, 65.14; H, 6,56;
N, 7.56 Actual value: C, 62,94; H, 6,
29; N, 7.51 benzyl (benzyl 2-acetamido-4,6-〇-benzylidene-2-deoxy-α-
D-glucopyranoside-6-O-yl)-acetyl-O
-Penzyru L-tononyl D-inglutaminate, melting point 182-188°C; [α)'',' + 76.
5゜L 10W #9/114. dimethylformamide)
, benzyl (benzyl 2-acetamido-4,6-〇-
Penzolidene-2-deoxy-α-D-glucot-lanosidol-6-〇-yl)-7cetyl-L-phenylalanyl-〇-inglutami*-), melting point 229° to 261°C
: [α)” + 78.6° (5■/-, dimethylformamide): Elemental analysis: C4aHsoN4011 (822-95
) Calculated values for: C, 65,68; H, 6,12
; N, 6.81 Actual value: C, 65,65; H
, 6,02; N, 6.62 benzyl (benzyl 2
-Acetamide-4#6-〇-benzylidene-2-deoxy-α-D-glucopyranoside-5-o-yl)-acetyl-〇-benzyl-L-tyrosyl-D-inglutaminate, melting point 241°-246°C; [α] Sweet 66.
7゜(4,2R9/d, dimethylformamide): benzyl (benzyl 2-7cetamido 4,6-0-benzylidene-2-deoxy-α-D-glucotlanoside 6
-〇-yl)-acetyl-ξ-carbobenzyloxy-L -1) Fluor D-isoglutaminate, melting point 215
° ~ 219 °C; [α] 25 + 66.7 ° (5,5r
n9/II/, dimethylformamide v): Elemental analysis:
Calculated values for C5oHsoNsO1s (958,06): c, 64.02; H, 6,54; N, 7
-47 Actual value: c, 66.79; H, 6,51;
N, 7-44 benzyl (benzyl 2-acetami?-
4. 6-〇-Benzylidene-2-deoxy-α-D-glucopyranosido-6-〇-yl)-7cetyl-L-prolyl-D-inglutaminate, melting point 224°-225
°C: [α)'', '+68.1° (,5,1!/-, dimethylformamide); Elemental analysis: Calculated value for 041H48N4011 (772,82): c, 66.72; H, 6,26 ;
N, 7.25 actual order: c, 65.49;
H, 6,39; N, 7.14 benzyl (benzyl 2-acetamido-4,6-0-benzylidene-2-
deoxy-α-D-glucopyranoside 6-0-yl)
-Acetyl-〇-benzyl-L-hydroxyzorolyl-D-isoglutaminate, melting point 219°-221°c:
[α]25+ 75.2° (9 da/d, dimethylformamide); Elemental analysis: C47H54N4011 (87
Calculated values for 8-99): C, 65, 59; H,
6,19; N, 6.67 Actual value: C, 65,49
; H, 6,42; N, 6.56 In a similar manner, benzyl 2-acetamido-4°6-0-benzylidene-2
-deoxy-6-o-(D-i-calphokidiethyl)-
α-D-1" for rucogiranoside), the compounds obtained in Examples 6.6' and 6" were used in stoichiometric equivalents and these were reacted with the appropriate compounds obtained in Example 4. , a variety of corresponding compounds of formula 00 are obtained, representative examples of which are shown below:
Benzyl 2-(benzyl 2-acetamido-486-〇-benzylidene-2-deoxy-α-ri-glucobylanosyl-'-5-0-yl)-D-butyryl-monovalyl D-inglutaminate, melting point 276°~ 276°
O; (a) 25 + 94.3° (1.4 m9/ml
, dimethylformamide); Elemental analysis: C43H5aN4011 (802,94
) total V# value for: c, 64.52; H, 6,
78; N, 6.98 Actual value: C, 64,00; H
,6,73; N, 6.81benzyl 2-(benzyl 2
-acetamido-4゜6-0-benzylidene-2-deoxy-α-D-glucopyranosige-6-〇-yl)-D
- detyruru L-prolylue D-inglutaminate,
[α]＾+87.6°(5m9/Il, shi)fruform7m)-″): Elemental analysis: C43HsgC43Hs (800,92
) Calculated values for: C, 64,48; H, 6,57
; N, 7.00 Actual value 2C, 64,20; H, 6,
52; N, 7.19 benzyl 2-(benzyl 2-
Acetamide-4゜6-o-benzylidene-2-deoxy-α-D-glucotlanosidol-6-〇-yl)-D-
hexanoyl-L-valyl-D-inglutaminate,
Melting point 264°~266°C; [α]25+ 76.6”
(6 da/-, dimethylformamide); Elemental analysis: C45HaaNaOtx (850-99
) <B calculation 1st shift vs. C: c, 65.04;
H, 7,04; N, 6.74 Actual value: c
, 65.26; ) (, 7, 27; N, 6.
81 Benzyl 2-(benzyl 2-acetamide-4゜6
-0-benzylidene-2-deoxy-α-D-glucopyranosyl-3-0-yl)-D-hexanoyl-L-zorolyl-D-ylglutamate, melting point 80°-88°C
; [α)” 85.7° (5 da/d, dimethylformamide); Elemental analysis: 045H456N4011 (828,9
8) Calculated value for K: c, 65-20; H, 6
, 81; N, 6.76 Actual value: c, 65.45;
H*6.86; N, 75.84 benzyl (benzyl 2-benzamide-4,6-〇-benzylidene-2-
deoxy-α-D-glucopyranoside-3-0-yl)
-7 cetyl L-valyl-D-inglutaminate, melting point 242°-249°C: [α]y 74.4° (5rn
9/d, dimethylformamide): Elemental analysis: C1aH5zkkO1z (856-95
) calculated values for: c, 66.01; H, 6,
26; N, 6.69 Actual value: C, 65, 82;
H, 6,20; N, 6.46penzyl 2-(benzyl 2-benzamido-4゜6-0-benzylidene-2-deoxy-α-D-glucobyranosido 6-o-yl)-
D-7' Robionyl-L-valyl-D-inglutaminate, melting point 257°-259°c:[α:]','+
73.2° (5 da/d, dimethylformamide): Elemental analysis: C47H54N4011 (850,98
) Calculated value for K: c, 66.64; H, 6,
40; N, 6.58 Actual value: c, 66-54 s
H-6,69s N -6,50benzyl 2-(benzyl 2-benzamide-4゜6-0-benzylidene-2
-deoxy-α-D-glucopyranoside 5-0-yl)-D-propionyl-L-prolyl-D-inglutaminate, melting point 1486-151°C; [α]','+<
50.8'45■/-, dimethylformamide): Elemental analysis: C4〒H5□N4011 (849,97)
Calculated value for: c, 66.49; H, 6,1
); N, 6.60 Actual value: c, 66.62;
H, 6,25; N, 6.58 benzyl (benzyl 2-
Calloamido-4,6-〇-benzylidene-2-deoxy-α-D-glucopyranosid-6-0-yl)-acetyl-L-valyl-D-inglutaminate, melting point 26
4°~265°C; [α)”+74.5° (5 da/r
nl, dimethylformamide): Elemental analysis: C41SH58N4011 (850-9
Calculated values for 9): C, 65,04; H, 7,0
4; N, 6.74 Actual value: C, 64,92;
H, 6,97; N, 6.74 benzyl (benzyl 2-
Caproamido-4,6-0-benzylidene-2-deoxy-α-p-glucopyranosid-6-〇-yl)acetyl-L-prolyl-D-inglutaminate, melting point 19
6゜～194℃z ('']+1'!l+ 58-0'
(5W/wl, y methylformamide): Elemental analysis: c, 5H, 6N, ol, (828,98
): c, 65.20; H, 6,8
1; N, 6.77 Actual value: c, 65.05; u
, 6.61; N, 6.85 Benzyl 2-(benzyl 2-caproamido-4゜6-0-benzylidene-2-deoxy-α-D-glucopyranosido 6-〇-yl)-
D-propionyl-L-valyl-D-inglutaminate, melting point 2520-256°C: [α]” + 86.7
°(5■/wLl, dimethylformamide), elemental analysis: C46H6ON4011 (845,02
): c, 65.68; H, 7,1
6; N, 6.66 Actual value: c, 65.66;
H, 6,99; N, 6.74 benzyl 2-(benzyl 2-caproamide-4゜6-0-benzylidene-
2-Deoxy-α-D-glucobyranosid 6-〇-yl)-D-propionyl-L-zorolyl-D-inglutaminate, melting point 1361'-168°C;(d)"δ
+86.0'(5'f/d, dimethylformamide), benzyl (benzyl 2-myrimethamide-4,6-〇-benzylidene-2-deoxy-α-p-glucopyranoside-6-〇-yl)-a7thyl L-valyl D-inglutaminate, benzyl (benzyl 2-myrimethamide-4,6-〇-benzylidene-2-deoxy-α-D-glucopyranoside 6-O-yl) -y*tyl-t.

-prolyl-low isoglutamate, benzyl 2-(
Benzyl 2-myristamide-4,6-0-benzylidene-2-deoxy-α-D-glucopyranoside 6-〇-
yl)-D-fropiony-L-valyl-D-isoglutamate, and benzyl 2-Lbenzyl 2-mi13stamiF-4゜6
-0-Benzylidene-2-deoxy-α-D-glucopyranosy P-,5-0-(L)-D-7'lobionyl-L-zorolyl-D-isoglutaminate.

Example 8 67fI acetic acid 45.0-
Benzylidene-2-deoxy-α-D-glucopyranosid-6-〇-yl)-〇-propionyl-L-valyl-D-inglutaminate (11, n = 2, Xi =
Add 5.9 g of L-valyl, R"=R2=C)I3). Heat the resulting solution to 97@-100 °C for exactly 7 minutes and then cool immediately in a water bath. Evaporate the solution under reduced pressure. After drying, the acetic acid is removed from the residue by double coevaporation with water and double coevaporation with toluene to obtain the crude product.

Ether 2501 was added to the product and the residue was diluted with ether 2501 at 22°C.
Stir magnetically for 5 hours. The resulting solid was filtered and dried,
Benzyl 2-(benzyl 2-acetamido-2-deoxy-α-D-glucopyranosido-5-o-yl)-D-
Propionyl-L-valyl-〇-inglutaminate (
12A, n-2, X'= L-baryl, R"=R6-
CM3) t-obtain: melting point 221-224°C: [a]
"p"106" 5° (10119/d, dimethylformamide): Elemental analysis: Cs5H4aNaO]1 (700,80)
Calculated values for: c, 59.99; H, 6,90
; N, 8.00 Actual value: c, 59.56; a
,6.92; )-D-7”-onyl-L-valyl-
Using stoichiometric amounts of the compound obtained in Example 7 in place of D-(soglutaminate) gives the corresponding compound of formula (12A), a representative example of which is shown below: benzyl (benzyl 2-acetamido- 2-Deoxy-α-D-glucopyranosyp-5-o-yl)-7cetyl-L-paryl-D
- Isoglutaminate, melting point 215°-222°C; [
α)”5+ 79.0° (I QIII9/mj, dimethylformamide): Elemental analysis: C34H46N40
11 (686,77) Calculated value for K; c, 59.
46; H, 6,75; N, 8.16 Actual value: c
, 59.56; H, 6,94; N, 7.92 cases
9 Benzyl 2-(benzyl 2-7cetamyP-2-deoxy-α-D-glucopyranosyl 1'-6-0-yl)-D
-Propionyl-L-valyl-D-isoglutaminate (12A% n=2, X'=L-valyl, R''=R
6=CH3) 0.701 and 0.56 g of triphenylmethyl chloride in 10-pyridine solution at 22°C for 5 days.
and then steam-dried under reduced pressure. The residue is partitioned between chloroform 5081 and water 50111, the chloroform phase is extracted two more times with 501 l of water, dried over magnesium sulfate, washed with ether, dried and the benzyl 2-(benzyl 2-acetamide- 2-deoxy-6-0-)liphenylmethyl-α-D-glucopyranosid-6-〇-yl)-D-propionyl-L-paryl-D-inglutamine) (12B, n=2, X'=L
-baryl, R2=R'=CH,) is obtained.

In a similar manner, benzyl 2-(benzyl 2-acetamido-2-deoxy-α-D-glucogyranosid 6-〇-yl)-D-! The compound obtained in Example 8 is used in a stoichiometric equivalent amount in place of Cubionyl-L-valyl-D-inglutaminate to obtain a compound of formula (12B).

Example 10 Benzyl 2-(Benzyl 2-acetamido-2-deoxy-α-D-glucopyranosido-6-0-yl)-D-
propionyl-L-paryl-D-isoglutamine)
(12A% n-2, X=L-valyl, R2=R6=C
H3) 0.56g, acetic anhydride 1, Qal Tonobi Q di: 1
The 5rnl@ solution is held at 22° C. for 16 hours, at which point 1 d of methanol is added and the resulting solution is stirred at room temperature for an additional hour. Solution M is then evaporated to dryness, the residue is partitioned between 201 Ll of chloroform and 20 L of water, the chloroform phase is dried over magnesium sulfate and then evaporated to dryness to obtain the crude product. The product was crystallized from aqueous ethanol and benzyl 2-(benzyl 2-acetamido-4,6-di-0-acetyl-2-deoxy-α-D
-Glucopyranoside 6-〇-yl)-〇-propionyl-L-paryl-D-isoglutamine) (12m,
n=2, X'=L-baryl, R"=R'=C
H3, R4 = R5 = C0CH5), melting point 22
5°~241°C: [α)”+95.7°(1011
qi/rtil, dimethylformamide): elemental analysis' 039H52N4013 (748,88)
Calculated value for VCN: c, 59.6B; H, 6,
68; N, 7.14 Actual value: C, 60,00;
H, 6, 76; N, 7.14 In a similar manner, a suitable acylating agent (acid anhydride or acid chloride) is used in a stoichiometric equivalent amount to O acetic anhydride, and the corresponding formula (12g) is obtained. Typical examples thereof are as follows; -7' Robionyl-L-valyl-D-inglutaminate, benzyl 2-(benzyl 2-acetamido-4゜6-di-0-octanoyl-2
-deoxy-α-D-glucopyranoside 6-〇-yl)-D-! Robionil-L-Vally-D-inglutaminate, melting point 207"-210°C: [α)" + 79
．． 6°C811 1) m1. dimethylformamide),
Benzyl 2-(bensyl 2-acetamido-4J6-di-o-stearoyl-2-deoxy-α-D-glucopyranoside 6-o-yl)-D-noropionyl-valyl-isoglutamate, melting point 200°~ 207°C, benzyl 2-(benzyl 2-acetamido-4゜6-di-O-pehenoyl-2-deoxy-α-D-curcobyranosid 6-〇-yl)-D-7'ropionyl-L-valyl-D - Inglutaminate, melting point 202-205°
O; (d)”+ 68.1° (5 m9/ml, dimethylformamide), and benzyl 2-(benzyl 2-7cetamido 4°6-di-0-benzoyl-2-deoxy-α-D-glucopyranoside 5-o-yl)-D-propionyl-L-valyl-D-inglutaminate, melting point 169° to 1) 2°C
.

In a similar manner, benzyl 2-(benzyl 2-7cetamido 2-deoxy-α-D-glucopyranoside 5-0
-yl)-D-propionyl-L-valyl-D-inglutaminate, the compound obtained in Example 8 is used in stoichiometric equivalent to obtain the corresponding compound of formula (12E),
Representative examples are shown below: benzyl 2-(benzyl 2-acetamido-4゜6-di-0-7cetyl-2-deoxy-α-D-glucopyranoside 6-〇-yl) -p -7' o hionyl L-prolyl-D-isoglutaminate, melting point 80°~
85°0; ((1)”+82.4° (10!/1, dimethylformamide): Elemental analysis: Cs*HaoN40x3 (782-86)
Calculated value for: c, 59.84; H, 6,4
4; N, 7.16 Actual value: c, 59.58
; H, 6,05; N, 7.13 pencil 2-(
Benzyl 2-acetamido-4゜6-di-0-butyryl-2-buoxy-α-D-glucopyranoside-6-0-
yl)-D-zologionyl-L-zorolyl-D-inglutaminate, benzyl 2-(benzyl 2-acetamido-4゜6-di-0-octanoyl-2-deoxy-α
-D-glucopyranoside 6-〇-yl)-D-propionyl-L-prolyl-p-isoglutaminate, benzyl (benzyl 2-acetamido-4,6-di-O
-acetyl-2-deoxy-α-D-glucopyranosido-6-〇-yl)-7cetyl-L-ba1) Ru p-
Isoglutaminate, melting point 197°-205°C; (a
]25+ 79.1' (10m97mls dimethylformamide): Elemental analysis' C3eHsoNaOx3 (770-85)
Calculated value for: c, 59.21; H, 6,5
4; N, 7.27 Actual value: c, 59.55;
H, 6,56; N, 7.28 Benzyl (benzyl 2-acetamiP-4, 6-di-O-butyryl-2-buoxy-α-D-glucopyranosid-6-〇-yl)-
7 cetyl 1°-valyl-D-isoglutamate, and benzyl (benzyl 2-7cetamido 4,6-di-0
-Octanoyl-2-deoxy-α-D-glucobyranosid 6-〇-yl)-acetyl-L-paryl-D-inglutaminate, melting point 1) 4 to 1) 7°C; r-a]
7 + 68.6° (8mp/mt.

dimethylformamide).

Example 11 Benzyl 2 (benzyl 2-acetamido-2--f,
t key/-α-D-glucopyranoside-6-〇-yl)-D-7'Robionyl-L-Ha! Benzyl 2-(pencyl 2-acetamido-2-deoxy-6-〇-triphenylmethyl-α-D-glucopyranosito-5-0-yl)-D-propionyl-L-valyl instead of J-Roo D-4 soglutaminate -D-isoglutaminate (1
2B s n ”' 2, XI: L-14 li/
l/, R''=R'=CH,) i Repeating the method of Example 10 but using stoichiometric equivalents, benzyl 2-(benzyl 2-acetamido-4-〇-acetyl-2-deoxy-6-0- ) liphenylmethyl-α-D-glucopyranosid-6-0-yl)-D-propionyl-L-valyl-D-isoglutaminate (12C, n=2, Xl=
L-baryl, R2=R'=CH3, R3=C0
CH5) is obtained.

In a similar manner, replacing acetic anhydride with a suitable acylating agent (either an acid anhydride or an acid chloride) in stoichiometric equivalents yields the corresponding compound of formula (12C).

In a similar manner, benzyl 2-(benzyl 2-amotoamido-2-deoxy-6-0-)! J phenylmethyl-α
-D-glucopyranoside-6-〇-yl)-〇-propionyl L-valyl D-inglutaminate is replaced by the compound obtained in 91J 9 in a stoichiometric equivalent amount,
The corresponding compound of formula (12C) is obtained.

Example 12 Benzyl 2-(benzyl 2-7 cetamide 4-〇-7
cetyl-6-〇-triphenylmethyl-α-D-glufuviranosido-6-〇-yl)-D-propionyl-L-
Valyl-D-isoglutaminate (12C1n=2,
L=L-baryl, R2=R'= CH3, R3= -C
OCH, ) 0.97 fi and 70% acetic acid 10- is heated at 100° C. until a clear solution is obtained (approximately 2 minutes). The solution is immediately cooled in a water bath and the precipitate is filtered off. The filtrate was evaporated to dryness under reduced pressure, the residue was mixed with ether 25d, the resulting solid was collected by filtration, washed with ether and dried under reduced pressure to give benzyl-2-(benzyl 2-
-7 Cetamita 4-〇-acetyl-α-D-glucopyranoside-5-o-yl)-D-propionyl-L-valyl-D-inglutaminate (12C, n=2, Xi
= L-baryl, R11=R6==CH3, R3=-
COCH3) is produced.

In a similar manner, benzyl 2-(benzyl 2-acetamido-4-o-acetyl-6-0-)liphenylmethyl-α-D-glucopyranosid-6-〇-yl)-D-propionyl-L-valyl- Using stoichiometric equivalents of the compound obtained in Example 11 in place of D-isoglutaminate, the corresponding compound of formula (12D) is obtained.

Example 16 Penzyl 2-(benzyl 2-acetamido-2-deoxy-α-D-glucopyranosido 6-〇-yl)-D-
Benzyl 2-(benzyl 2-acetamide-4
-0-7 cetyl α-D-glucopyranoside-6-0-
Il)-〇-propionyl-L-valyl-D-inglutaminate (12D% n=2, Xi = L-valyl, R2==R' = CH3, R' = -COC'H3
) is used in stoichiometric equivalents and propionic anhydride is used in place of acetic anhydride,
benzyl 2-(pairzyl 2-acetamido-4-〇-acetyl-6-0-f robionyl-α-D-/rucobyranosidol-6-〇-yl)-D-progionyl-L-valyl-D-inglutamine) ( 12E.

n=2, X1=L-baryl, R"=R6=CH3
, R''-COC2H3, R'=-C'OC2H5).

In a similar manner, using a suitable acylating agent (acid anhydride or acid chloride) in a stoichiometric equivalent amount in place of propionic anhydride), the corresponding compound of formula (12E) is obtained.

In a similar manner, benzyl 2-(benzyl 2-acetamido-4-〇-acetyl-α-D-glucopyranoside-6
The compound obtained in Example 12 is used in stoichiometric equivalents in place of -D-propionyl-valyl-D-isoglutaminate to give the corresponding compound of formula (12E).

Example 14 Benzyl 2-(benzyl 2-acetamido-2-deoxy-α-D-glucopyranosido-6-0-yl)-D-
Propionyl-L-valyl-D-isoglutaminate (
12A, n=2, X''=L-baryl, R8=R'=C
H3) Add 0.66 g of gin 5 solution dropwise over 60 minutes with acetic anhydride Q, Q 95 m/birisin 1 and stir at 0°C. The resulting solution is held at 0° C. for 16 hours, at which point 1 g of methanol is added and the mixture is evaporated to dryness. The residue thus obtained is partitioned between 100 mj of chloroform and 100 mj of water, the chloroform phase is dried over magnesium sulfate and concentrated under reduced pressure, yielding 0.56 fi of crude product. This product was purified by preparative thick layer chromatography and benzyl 2-
(Benzyl 2-acetami)-'-6-0-acetyl-
2-Deoxy-α-D-glucopyranosido-6-〇-yl)-D-propionyl-L-valyl-D-isoglutaminate (12F.

n=2, X''=L-baryl, R2=R'=CH3
, R3=-COCH3).

In a similar manner, a suitable acylating agent (#
R anhydride or acid chloride) in stoichiometric equivalents to give the corresponding compound of formula (12F).

In a similar manner, benzyl 2-(benzyl-2-acetamido-2-deoxy-α-D-glucopyranoside 6-
Using the chemical equivalent of the compound obtained in Example 8 to (l)-D-propionyl-L-valyl-D-inglutaminate (12F) gives a compound of formula (12F).

Example 15 Benzyl 2-(benzyl 2-acetamide-4゜6-0
-benzylidene-2-deoxy-α-D-glucopyranosido-6-0-yl)-D-7'ropionyl-L-valyl-D-inglutaminate (11, n = 2, Xl
= L, -baryl, R2== R15==CH3) 0
．． A mixture of 5011 and 50 vv of palladium on 10% carbon catalyst in 15 m of 85% acetic acid was heated to 22 m in a hydrogen atmosphere.
Stir magnetically at °C. Product (16, n = 2,
X=L-valyl, R”=R6=CH3) is formed by a
Follow-up by thin layer chromatography using a solvent base of acetonitrile:acetic acid:water in the ratio:i:1. It can be seen that this reaction is generally complete in 24-40 hours. At this point, the catalyst is removed by filtration through a microporous filter (Teflon) and the filtrate is evaporated to dryness under reduced pressure to yield a residue. The residue is dissolved in 6-methanol and the product is then precipitated by addition of 8 ml of ethyl acetate. The white solid thus obtained is collected by centrifugation and carefully dried under reduced pressure to obtain a white powder. This powder is dissolved in 5 ml of water and the solution is then passed over a 20 cm xo, 5 cI11 column of Biolex o (B10 order) 70 (a weakly acidic ion-exchange resin of polyacrylic acid). The column was eluted with water and the fractions containing the desired product were collected, lyophilized in vacuo and 2-(2-7cetamido-2-deoxy-D-glucose-6-〇-yl)-D-propionyl -L-valyl-D-isoglutamine (16, n=2, M); ((ri'W+15
．． <5'(10'9/-, methanol); Elemental analysis: Cs+1H3aN401x "R20(5
58-57) VC vs. f Le calculated value: C, 46,85
; a, 7.11; N$ 10.40 actual fia
t: C, 46,66; H, 7,26; N, 1
0.2<S In a similar manner, benzyl 2-(benzyl-2
-acetamido-4,6-0-benzylidene-2-deoxy-α-D-glucopyranosido-5-0-yl)-D
In place of -7'-onyl-L-valyl-D-isoglutaminate, various products from Example 7 are used in stoichiometric equivalents to give the corresponding glycopeptides of formula 0. Representative compounds thus obtained are shown below: 2-(2-acetamido-2-deoxy-D-glucose-6-0-yl)-D-7'ropionyl-L-phenyl7ranyl-D-iso Glutamine, melting point 140°~14
6℃; [α]22+ 84.4° (5.9 Da/1, methanol); Elemental analysis: C25H36N4011・1 l/2 R
20 (595-62) Calculated value for Ic: c, 50.41; H, 6,60; N
, 9.41 Actual value: c, 50.25; H, 6
, 61; N, 9.27 In a similar manner, benzyl 2-
(Pencil 2-7cetamido 4.6-0-benzylidene-2-deoxy-α-D-glucogyranidoside 5-
In place of 0-yl)-D-propionyl-valyl-D-inglutaminate, the various products from Example 10 are used in stoichiometric equivalents to give the corresponding compounds of formula 03. Representative compounds thus obtained are shown below: 2-(4,6-di-0-acetyl-2-acetamide-
2-deoxy-D-glucose-5-0-yl)-D-
Propionyl-L-valyl-D-isoglutamine; melting point 150° ~ 1) 0'C (decomposition): [α stomach + 28.6° (
10 Da/d, methanol); Elemental analysis: C25H4O
Calculated value for N4013 2H20C640,66): c, 46.87; H, 6,92; N
, 8.75 Actual value: c, 46.96; H, 6,22
; N, 8.612-(4,6-di-0-octanoyl-2-acetamyl-2-deoxy-D-glucose-
6-〇-yl)-D-propionyl-L-valyl-D-
Inglutamine, melting point 198°-200°C: (d stomach +
45.9° (5r!9/I-(, mep 7-A/)
;Elemental analysis' C3? H64N40□3 (772,9
Calculated values for 5): c, 57.49; H, 8
, 55; N, 7.25 Actual value: c, 57.52
; H, 8,27; N, 7.262-(4,6-di-
0-stearoyl-2-acetamido-2-deoxy-
D-glucose-60-1)-D-7'rohionyl-monovalyl D-isoglutamine; melting point 187°C12
-4,6-di-O-behenyl-2-acetamide-2
-deoxy-D-glucose-6-〇-yl)-D-7
'Robionyl-L-valyl-D isoglutamine; melting point 1
) 5° to 185° Cl2-(4,6-di-0-benzoyl-2-acetamido-2-deoxy-D-glucose-6-0-yl)-D-propionyl-L-valyl-D
- Isoglutamine: melting point 152 ° G (decomposed), obtained by formula 0 [and/or) (1 ) is divided into its individual Oα- and β-anomers.

Example 16 20 μl of tyrosine arsanilate and protein [either bovine serum albumin (BSA) or ovalbumin]
An antigen mixture consisting of 100 μg is used. This antigen mixture and the test compound are dissolved in saline in 1 guinea pig doses shown in the table below and emulsified with equicrine Freund's incomplete adjupan (CFIA). Groups of 10 guinea pigs were used for each test compound unless otherwise specified;
Each guinea pig is injected subcutaneously with 0.1 of the above emulsion in the eel area. In all analyses, the control group received the antigen mixture emulsified with FIA, and in most analyses, the test compound was also administered with 2-(2-7cetamido-2-deoxy-D-glucose-6-o-yl)-D -Propionyl-
Compare with L-alanyl-D-isoglutamine under the same conditions.

After 14 days, 25 μg of tyrosine arsanilate conjugated to BSA or ovalbumin was injected intradermally into the flank of the animals in 0.1 l saline and all animals were subjected to skin testing using a skin test method. (BOA is used in this test if ovalbumin is present in the antigen mixture, and ovalbumin is used vice versa). A number between 1 and 6, which is a measure of the size of the infiltration observed in each skin test, stimulates a delayed hypersensitivity reaction to tyrosine arsanilate by multiplying the average area of skin reaction at 24 hours. Calculate the potency of the test compound. Thus, the expression is as follows: skin tests showing a high degree of infiltration are recorded with a ++ and given the value 6; those showing a lesser degree of infiltration are marked with a 10 and given the value 2.
t-gives; on the other hand, skin reactions that do not show hydration are represented by 0 and assigned a value of 1. These values are then summed for a group of animals and the resulting average value is multiplied by the average area of observed skin response. This titer thus encompasses the degree of penetration combined with the average area observed in the skin test. The average titer of the test compound is shown in column 1 of the following table and compared to the FIA having a value of 1.

On day 42, all animals were bled by cardiac puncture.
Farr's technique using labeled bovine serum aldemi/[P. Minden & R. Farr.
Handbook of Experimental Immunology
talImmunology), Day War Jl-A
#Wire (D, M.

Weir) published, Chapter 15, Volume 1, Blackwell Scientific Research, Oxford University Press, by radioimmunoassay, or by passive hemagglutination, the serum was treated with the antibiotic serum aldemin or by passive hemagglutination. Analyze for anti-ovaldemin antibodies. The average titers for these assays are shown in column 6 of the following table, compared to the FIA, which has a value of 1.

On day 42, all surviving animals undergo a skin test with 10 μg of protein (BOA if BOA is present in the antigen mixture or ovalbumin if vice versa). The potency of the test compound to stimulate a delayed-type hypersensitivity reaction to this protein compared to an FIA with a value of 1 at 24 hours was determined exactly as described above for the delayed-type hypersensitivity reaction to tyrosine arsanilate. and is shown in the second column of the following table.

Table annotations: a = 0.1 μmol dose.

c=Relative potency to the potency of 2-(2-7cetamido-2-deoxy-D-glucose-3-0-yl)-D-propionyl-L-7ranyl-D-isoglutamine 20 μl dose.

θ-Whether skin test or body fluid antibody synthesis λ F Tech A
The actual response to FOA can vary greatly compared to the response obtained with either FOA or test compounds given with FFA. This reflects the high degree of variability in titers calculated by calculating the mean titer of the test compound relative to the mean titer of the FIA control group in the same analysis.
t 41 extra. Therefore, we calculated the potency of the test compound as follows: 2-(2-
Median titer of acetamido-2-deoxy-D-glucose-6-〇-yl)-D-f robionyl-L-7 ranyl-D-isoglutamine t-#1 added and 2-(2-7 Cetamido-2-deoxy-D-glucose-6-〇-yl)-rz-f robionyl-L-
722-(2-acetamido-2-deoxy-D)
-glucose-6-〇-ilun-D-progionyl-L
The intermediate potency of -alanyl-D-inglutamine and the multiplication factor are compared with R-Technique A, which has the number 1 of the potency t1 of the test compound, calculated by calculating.

Example 1) Acute toxicity test method in mice = 0.9% sodium chloride, 0.5% sodium carboxymethylcellulose, 0.4% polysorbate)
The test substance was dissolved or homogeneously suspended in an aqueous medium containing 80 and 0.9% P/D alcohol A/f.

10-7 The concentration so that the indicated dose can be administered in the amount of 9 to 11? ! - Adjustment friend. Next, 6 male Swiss-Webster mice in the test substance kI group were given 60 da/kl of body weight. The number of deaths of the animals was observed daily for 21 days after a downwind injection (8,0,) in a holding shed. A control group of mice not receiving the drug was challenged at the same time. As can be seen from the accompanying table, all of the tested compounds showed LD, o values greater than 60 Da/11 Yu. This particular dose dose is approximately 60% of the amount required to elicit an enhanced delayed-type hypersensitivity and to circulate the antibody levels described in the example.
It was selected because it corresponds to 0 times.

(continued)

Claims (10)

[Claims] (1) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [In the formula, R and R^1 are the same and represent hydrogen, acetyl, octanoyl, stearoyl, behenoyl, or benzoyl; R^2 represents methyl; R^3 represents methyl; When one of the wavy lines is α, the other is β; provided that when R and R^1 represent hydrogen, X is not L-valyl]. (2) The compound according to claim 1, wherein R and R^1 are each hydrogen. (3) The compound according to claim 1, wherein R and R^1 are acetyl, octanoyl, stearoyl, behenoyl, or benzoyl groups. (4) The compound according to claim 1, wherein X is L-valyl. (5) R and R^1 are both acetyl, and
2. A compound according to claim 1, wherein is L-valyl. (6) The compound according to claim 1, wherein R and R^1 are both octanoyl, and X is L-valyl. (7) The compound according to claim 1, wherein both R and R^1 are stearoyl, and X is L-valyl. (8) R and R^1 are both acetyl, and
is L-valyl; i.e. 2-(4,6-di-O-
The compound according to claim 1, which is acetyl-2-acetamido-2-deoxy-D-glucose-3-O-yl]-D-propionyl-L-valyl-D-isoglutamine. (9) Formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R and R^1 are the same and represent hydrogen, acetyl, octanoyl, stearoyl, behenoyl, or benzoyl; R^2 is methyl. Representation; R^
6 represents methyl; When one of the wavy lines is α, the other is β; however, when R and R^1 represent hydrogen, X is not L-valyl] A method for producing a compound represented by the formula ▲ There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R', R^1', R^2' and X' are the groups R, R^1, R^2 and X of the above formula I, respectively,
Z is hydrogen and P is hydrogen, provided that R', R^
1', R^2', X', Z and at least one of P is a protecting group]. (10) The method according to claim 9, wherein the protecting group to be removed is selected from the group consisting of benzyl ester, benzyl ether, benzylamino, or benzylidene acetal.