JPH0371440B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the formula: Regarding the novel immune adjuvant compound shown in
Ru. In the above formula, R and R¹are the same
, acetyl, octanoyl, stearoyl,
Represents henoyl or benzoyl; R²Ha mechi
Represents R⁶represents methyl;
represents ril; Y represents D-isoglutamine
and the wavy line (〓) indicates α- or β-coordination,
Rui also represents a mixture, but one of the wavy lines
is α, then the other is β. For the sake of brevity in the reaction route diagrams described below,
In addition, to avoid excessive redundancy, in the above formula
of The part of the formula that corresponds to the part shown in i.e. has the subformula shown in formula (b)
The compound shown in formula (a) is actually a steric compound of the subformula shown in formula (a).
It is understood that it has an arrangement structure. X is L-valyl and Y is D-isoglu
Particularly preferred are compounds of the formula tamine. In the field of immunology, viruses (or
bacteria) that elicit a sufficient host immunological response to prevent infection.
One or more vaccines (or vaccines) must be used to
injections of Cterin) are usually required. This is the trick
Virus contained in chin (or bacterin)
(or bacterial) antigen to the animal (or human) host
It is purified very quickly from the place where it was injected,
When the host is insufficient to mount its own immune response
This is because there is only time. Delays the release of viral (or bacterial) antigens
and also to integrate the host's immunological system.
Vaccines (or bactericides) are commonly used to stimulate
phosphorus) with an immune adjuvant and multiple injections
Efforts are being made to eliminate the need to do so. death
However, these conventional immune adjuvants
For example carrion, tragacanth, bentonite
Carbopol, Calcium Phosphate Gel, Tapi
Oca, alum, aluminum hydroxide, calci chloride
Many of the drugs, such as alginate and sodium alginate, are injectable.
are non-metabolic when exposed to
The adjuvant itself has the disadvantage of being a source of stimulation. It can be easily metabolized by the host and
sometimes slows down the release of antigen and generalizes the immune response.
Immune adjuvant substances that stimulate
It was highly sought after. Immune adjuvants commonly used in laboratories are (a)
Death of all Mycobacterium tuberculosis
Fluids are suspensions of bacteria in mineral oil and emulsifiers.
Roint fully adjuvant (FCA); and (b) ore.
Freund incomplete ajiyuba with only oil and emulsifier
(FIA). FCA and FIA are laboratory standards
Although used as quasi-substances, these are not commercially available.
is not used. This is because FCA is a toxic
is derived from the organism, Mycobacterium tuberculosis;
FIA is highly sufficient in the absence of dead M. tuberculosis.
This is because it does not produce an immunological response. Ellouz et al.
and biophysical research community
NICATIONS (Biochemical and Biophysical
Research Communications), Vol. 59, No. 4, 1317
~1325 (1974), some species obtained from microorganisms
Peptidoglycan has immune adjuvant activity.
and used in place of all killed Mycobacterium tuberculosis in FCA.
(Also, Belgian patent no.
(See also No. 821385). For example, the novel peptidoglycan of formula () is
Bovine serum albumin (BSA) or ovalbumin
Antibody titer (antibody titer) against either
significant immune adjuvant, as measured by titres)
[D.J. Herbert]
(W.J.Herbert), Ch.20, Volume 1, Handbook
of experimental immunology
(Handbook of Experimental Immunology),
Editor
D.M.Weir), Blackwell Scientif
Tsuku Publications, Oxford
(Blackwell Scientific Publications, oxford)
(1973)] and described in detail in Example 16.
So, either BSA or ovalbumin
delayed-type hypersensitivity to
Liei and R.G. White (J.H.
Humphrey and R.G.White), Immunology Foundation
Students of Medicine
(Immunology for Students of Medicine), 493
~545 pages; Publisher F.A. Davis, Inc. (F.A.
Davis Co.), Philadelphia, 3rd edition (1970)
], or delay to tyrosine arsanilate
Type hypersensitivity [S. Laskowitz,
J.Exp.Med., 119,
291 (1963) and S. Laskowaits Scien.
Science, 155, 350 (1975)].
Furthermore, the compound of formula
Vaccine (or bacterin) compositions containing
Mineral oil and vegetable oil
When used in vaccines (or bacterins) containing
It does not exhibit the harmful side effects normally observed in other cases. Furthermore, the compound of formula may itself be infectious.
Aircraft, such as Klebsiella
pneumoniae), Escherichia coli,
Candida albicans, or yellow bud
Activity against Staphylococcus aureus
have sex. New compounds of the formula are generally shown below.
It can be produced according to the reaction route: In each of the above formulas, R, R¹,R²,X and
(〓) is as defined above; R³is acyl having 1 to 22 carbon atoms
is the basis; R^Fourand R^Fiveis the same or different 1 to 22
is an acyl group having carbon atoms; R⁶is hydrogen or has 1 to 16 carbon atoms
is an alkyl group; R⁷is hydrogen or methyl; n is an integer of 1 or 2; Ph is phenyl; and X¹is glycyl, L-alanyl, L-valyl,
L-leucyl, L-isoleucyl, L-α-ami
Nobutyryl, O-benzyl-L-seryl, O-be
ndyl-L-threonyl, L-methionyl, S-
Benzyl-L-cysteinyl, L-phenylara
Nyl, O-benzyl-L-tyrosyl, L-trip
Tofa, ε-carbobenzyloxy-L-lysi
L, δ-carbobenzyloxy-L-ornithyl
gu-nitro-L-arginyl, N^Im-Venji
-L-histidyl, γ-ben of L-glutamine
Dyl ester, L-glutaminyl, L-aspar
β-benzyl ester of chill, L-asparagini
L-prolyl, L-prolyl, or O-benzyl-L-hydroxy
selected from the group consisting of droxyprolyl. A compound of formula (13) in which n is an integer of 1 or 2
In this case, the compound thus obtained is such that Y is
D-isoasparagine or D-isoglu, respectively
It turns out that Tamin is a compound with the formula
Dew. When carrying out the above method, D-amino of formula (1)
Benzyl alcohol, P-methoxybenzyl acid
Alcohol, P-nitrobenzyl alcohol, etc.
aralkyl alcohol, preferably benzene
alcohol, hydrogen chloride, P-toluene sulfonate.
in the presence of a strong acid such as phosphoric acid, preferably sulfuric acid,
At a temperature of about 0° to about 40°C, preferably 20 to 25°C
treatment for about 8 to about 40 hours, preferably 14 to 18 hours.
A compound of formula (2) is obtained. The amino ester of formula (2) thus obtained
group, which is well known in polypeptide synthesis technology.
and generally acyl (e.g. formin, triflu)
(olacetyl, phthaloyl, etc.); urethane (and
For example, t-butyloxycarbonyl, P-methoxy
Benzyloxycarbonyl, 2-(P-bipheni)
-isopropyloxycarbonyl, isoniyuti
sulfenyl (O-
Nitrophenylsulfenyl, tritylsulfenyl
or alkyl (triphenylmethyl,
benzhydryl, etc.).
protection by blocking with a removable group.
More particularly, an appropriate reaction with an amino ester of formula (2)
agents (e.g. t-butyl fluorate, azide
t-butyl formate, t-butyl phenyl carbonate, etc.
(preferably t-butyl azidoformate) in an aqueous-organic
in mixtures or anhydrous organic solvents (e.g. dioxa
Tetrahydrofuran, Dimethylformamide
dimethyl sulfoxide, preferably anhydrous dimethyl sulfoxide, etc.
methyl sulfoxide), a base (e.g. hydroxyl
sodium chloride, magnesium oxide, tetramethyl
Guanidine, treethylamine, etc., preferably treethylamine, etc.
from about 12 to about 48 hours.
from about 0° to about 40°C, preferably for 20 to 24 hours,
Preferably, by reacting at 20 to 25°C, formula (3)
protected amino ester, N-t-buty
A carbonyl derivative is produced. The carbo of this protected amino ester of formula (3)
One of the commonly used reagents for the formation of amide bonds with xyl groups
(e.g. dicyclohexylcarbodiimide, dicyclohexylcarbodiimide,
Isopropylcarbodiimide or other carbodiimide
Diimide to N-hydroxysuccinimide or
1-Hydroxybenzotriazole; Chlorformic acid
isobutyl or other chloroformate;
2-ethyl-5-(3-sulfophenyl)-1,2
-oxazole-betaine or other 1,2
-Oxazolium salt; thionyl chloride or pentachloride
Phosphorus; 2,2'-dipyridyl disulfide/tri
phenylphosphine; etc., preferably chloroformic acid
with or using additives such as isobutyl.
(without using) and then anhydrous
Organic solvents (e.g. dioxane, tetrahydro
in furan, preferably tetrahydrofuran)
About 30 minutes to about 12 hours at -20° to about 40°C,
Preferably at about -15° to about 25°C for 1 hour.
By treatment with anhydrous ammonia, the compound of formula (4), all
That is, β-benzyl t-butyloxycarbonyl
-D-isoaspartate (n=1) or γ
-benzyl t-butyloxycarbonyl-D-i
Generate soglutamate (n=2). As shown above for t-butyloxycarbonyl
When N〓-protecting groups are well known to those skilled in the art,
Suitable methods for the group of protecting groups used
(e.g. P-toluenesulfonic acid, methanesulfonic acid)
Honic acid, hydrogen chloride, trifluoroacetic acid, formic acid, trifluoroacetic acid
Boronide or other moderate strength protons
an inert organic solvent by the donor or electron acceptor;
Preferably in a saturated solution of hydrogen chloride in ethyl acetate.
for about 1 to about 60 minutes at about -15° to about 40°C
between about 15°C and about 25°C, preferably between about 15°C and about 25°C.
Treat for about 30 minutes, then add a solvent with a low dielectric constant,
For example, hexane, ether, benzene, etc.
(or diluted with ether) to protect formula (4).
isoasparagine β-benzyl ester or
Is it an isoglutamine γ-benzyl ester compound?
is removed from the compound of formula (5), i.e. β-benzi
lysoasparaginate hydrochloride (n=1) or
γ-Benzyl isoglutamate hydrochloride (n=
2) is generated. Next, the t-butyloxycarbonyl compound of formula (6)
react with a compound of formula (5) to obtain a compound of formula (7).
Ru. This reaction is carried out in polar solvents (ethyl acetate, acetonate).
Tolyl, tetrahydrofuran, dimethylformua
salt in dimethylformamide, preferably dimethylformamide).
groups (e.g. triethylamine, diisopropyl
Ethylamine, N-methylmorpholine, etc., preferably
is about - in the presence of diisopropylethylamine)
Preferably at 10° to about 80°C for about 30 minutes to about 4 hours.
Preferably at about 10° to about 25°C for about 30 minutes to about
The reaction was carried out for 1 hour using a suitable additive (e.g. formula (4)
For the addition of the auxiliary agents mentioned above for the production of the compound of
, n=1 or 2 and X¹
is as defined above.
Generate petide. Alternatively, as shown in equation (6),
Other instead of t-butyloxycarbonyl group
Regarding the production of N-protecting groups, e.g. compounds of formula (3)
Groups such as those mentioned above can also be used. N-acetyl glycopeptide of formula (13) (R=R¹
=H,R²＝CH₃, n=1 or 2, X and R⁶
is as defined above) is the equation (8) to (13).
Follow as illustrated in the reaction pathway to produce
can get. The raw material for this reaction route is the benzyl 2-
Acetamide-4,6-benzylidene-2-deo
xy-α-D-glucopyranoside, and this compound
The compound is manufactured by P.H.Gross and Art.
P.H.Gross and
R.W.Jeanloz)
(J.Org.Chem.), 32, 2762 (1967)
There is. All of this continuous process is β−, which corresponds to equation (8).
Equivalent even if benzyl glycoside is used as a raw material
The synthesis of this starting material can also be performed in a die-cutting manner.
Lee Org Chemi. 32, 2762 (1967)
ing. These steps are performed using α-benzyl glycol of formula (8).
Described in the case of using coside as a raw material
All under conditions generally comparable to those of Alkylation of the free hydroxyl group in formula (8) is
Such as thorium hydride, potassium hydride, etc.
Using an appropriate base, dimethylformamide,
Inerts such as trahydrofuran, dioxane, etc.
About 30 minutes to about 0° to about 50°C in an organic solvent
for about 3 hours, preferably in dimethylformamide.
Process at 10° to about 30°C for about 15 to about 60 minutes.
easily by first forming its anionic form.
It can be achieved. This anionic body is then preferably
having 2 to 18 carbon atoms in the same solvent
Straight-chain α-halocarboxylic acid (typically chloroacetic acid)
acid, α-chloropropionic acid, α-bromopropionic acid
onic acid, α-chlorobutyric acid, α-bromobutyric acid, α-
Bromovaleric acid, α-bromocaproic acid, α-bromo
mumiristic acid and α-bromstearic acid
). This process requires dimethylform
in amide at about 20° to about 100°C for about 15 minutes or
about 5 hours, preferably about 0.5 at about 50° to about 80°C
for about 2 hours or at about 20℃ to about 30℃
Advantageously, it is carried out for 10 hours to about 48 hours. Equation (9) to generate (R²＝CH₃,R⁷=H,R⁶= the above
The compound (as defined) is then conventionally collected
isolated and crystallized by methods known to those skilled in the art.
Purify by other methods as described. Alternatively, a compound of formula (9) (R²＝CH₃,R⁷=
H,R⁶= as defined above) into equation (9)
The corresponding methyl ester of the compound (R²=R⁷＝CH₃)
by conventional methods, and then by conventional methods, e.g.
By crystallization or chromatography on silicic acid
Purification is preferred. This methyl ester
The production method is to use the acid of formula (9) (R²＝CH₃,R⁷=H)
Azomethane, methanol, and tetrahydrofura
dioxane, etc., preferably tetrahydrofuran
at a temperature of about 5° to about 40°C, preferably about
About 5 minutes to about 2 hours at a temperature of 20° to about 25°C
The reaction time is preferably about 15 to about 30 minutes.
compatible with methyl iodide or dimethyl sulfate.
suitable bases (e.g. potassium carbonate, sodium hydroxide)
solvent (e.g., triethylamine, etc.) in the presence of
For example, tetrahydrofuran, dioxane, dimethyl
formamide, etc.) at about 5° to about 50°C for about 30 minutes.
This includes a reaction in which the reaction is carried out for about 16 hours. suitable
This esterification is carried out using water and dimethyl sulfate.
about 20° to about 25°C following continuous addition of ate.
By reacting for about 14 hours to about 18 hours with
Performed in situ in the alkylation medium. In another embodiment of this method, the free hydrochloride of formula (8)
The alkylation of the xy group was carried out as described above.
The anionic form with 2 to 18 carbon atoms
straight-chain α-halocarboxylic acids (typical examples include chlorocarboxylic acids)
Methyl acetate, methyl 2-bromopropionate,
Methyl 2-bromobutyrate, Methyl 2-bromocaproate
methyl, 2-bromocaprate, 2-bromo
Methyl myristate and 2-bromostearin
methyl acids) and esters (e.g. methyl, ethyl
(chill, n-propyl, etc.)
achieve. This method is R⁶is 5 to 16 carbons
A compound of formula (9) which is a straight chain alkyl group having an atom
It is particularly useful for the production of formula (9) (R²=R⁷＝CH₃)
It is advantageous to directly produce the methyl ester of
Therefore, this methyl ester of formula (9) is crystallized and crystallized on silicic acid.
chromatography or as known to those skilled in the art.
Purify by other methods. This reaction is dimethic
Formamide, Tetrahydrofuran, Dioxa
at 0° to 100°C in an inert organic solvent such as
for about 15 minutes to about 24 hours, preferably dimethic acid.
in formamide at about 10° to about 30°C for about 30 minutes
Do this for at least 2 hours. In some cases, linear α-ha
(8) in a stirred solution of an appropriate ester of a carboxylic acid.
Drop a dimethylformamide solution of the anionic substance of
A high yield of the compound of formula (9) can be obtained by adding
can get. The compound of formula (9) (R²=R⁷＝CH₃) is then mixed with water.
suitable organic solvents (e.g. methanol, ethanol,
Dioxane, tetrahydrofuran, etc., preferably methane.
20°C or more in a mixture with (tanol)
Reflux temperature of the medium mixture, preferably about 60° to 90°C
for about 10 minutes to about 4 hours, preferably about
Add a suitable base (e.g. sodium hydroxide) for 15 minutes to 1 hour.
Thorium, potassium hydroxide, barium hydroxide, etc.
hydrolyzed, preferably using sodium hydroxide).
The corresponding carboxylic acid (R²=
CH₃,R⁷=H). The compound of formula (9) (R²＝CH₃,R⁷=H) and equation (10)
Various suitable protected benzyl α-aminoacyl
Ru-D-isoglutamate and benzyl α-
Aminoacyl-D-isoaspartate derivative
(X in the formula¹is glycyl, L-alanyl, L-vari
Le, L-leucyl, L-isoleucyl, L-α-
Aminobutyryl, O-benzyl-L-seryl, O
-benzyl-L-threonyl, L-methionyl,
S-benzyl-L-cysteinyl, L-phenyl
Alanyl, O-benzyl-L-tyrosyl, L-t
Lyptophanil, ε-carbobenzyloxy-L
-lysyl, δ-carbobenzyloxy-L-ol
Nityl, gu-nitro-L-arginyl, N^Im-be
γ- of L-histidyl, L-glutamyl
Benzyl ester, L-glutaminyl, L-as
β-benzyl ester of partyl, L-aspara
Guinyl, L-prolyl and O-benzyl-L-
hydroxyprolyl and n=1 or
The condensation with 2) requires a person familiar with peptide synthesis technology.
performed by various techniques well known to those who
Now. That is, for example, dipeptide hydrochloride of formula (10)
Salt derivatives such as diisopropylethylamine
The free salt can be prepared by adding a suitable strongly basic tertiary amine.
Convert to base form. Formation corresponding to the compound of formula (10)
Next, the free amino compound of formula (9) (e.g.
BaR²＝CH₃,R⁷=H and R⁶is as defined above
carbonyl group) and dicyclo
Hexylcarbodiimide or diisopropylcarbohydrate
using a carbodiimide such as rubodiimide
Dimethylformamide, dichloromethane, acetic acid ethyl
Non-containing substances such as dioxane, acetonitrile, etc.
Condensation in an active organic solvent. Depending on the case, also
Preferably, this reaction is carried out using 1-hydroxybenzotri
Azole, N-hydroxysuccinimide, P-
Nitrophenol, pentachlorophenol, etc.
In the presence of suitable additives well known such as
Let's do it. According to the invention, in dimethylformamide
For about 2 to about 60 hours at about 0° to about 50°C, preferably
Suitably at about 10° to about 30°C for about 10 to about 40 hours,
Diisopropylcarbodiimide and 1-hydro
Excellent condensation using xybenzotriazole
found to yield results. Generate(13)
The product of formula (R⁶,X¹and n as defined above.
crystallization and quenching on silicic acid.
Purified by conventional methods including romatography
do. Alternatively, a free base of formula (10) and a compound of formula (9)
Things (R⁷=H) can also be easily condensed with peptide technology.
It can also be done by other well known methods.
That is, for example, chloroformate (e.g.
isobutyl chloroformate) or 1-ethoxylic
Bonyl-2-ethoxy-1,2-dihydroquinolite
The mixture derived from the compound of formula (9) in the presence of
Using water products is effective. Also, for example, Zipi
Lysyl disulfide or carbon tetrahalide
triphenylphosphine or oxa in the presence of
Another problem caused by the use of zolium salts is
Carbonyl activation of ip is also useful. like this
A general overview of condensation methods is provided by I.S.
Klausner and M. Bodansky (I.S.Klausner)
and M.Bodansky), Synthesis,
453 (1972) and G.H.
(J.H.Jones), Chemistry and Indus
Torii (Chemistry and Industry), 723 (1974)
found in the paper. (8) to (9) (R⁷=H) Reaction sequence that proceeds to
is also an N-acyl functional group, R²Change in the properties of CO−
can also be changed in such a way as to make it possible to
Ru. That is, the N-acetyl functional group in formula (8) is
J.Org.Chem., 32, 2762
(1967) by hydrolysis with a strong base,
Benzyl 2-amino-4,6-benzylidene-2
-deoxy-α-D-glucopyranoside, formula
(8A) can be generated. This method is
Also described in Lee Org Chemi, 32, 2762 (1967)
Apply to the β-anomer of formula (8) as shown in
It can also produce benzyl 2-amino-4,
6-Benzylidene-2-deoxy-β-D-glue
Copyranoside is related to the reaction starting from equation (8A).
and subsequent reaction under essentially the same conditions as those described.
Can be used as appropriate. Selective N- of the compound of formula (8A)
Acylation is performed using lower alcohols, preferably methanol.
produced by reacting with carboxylic anhydride in a
wake up This acid anhydride is a linear and branched aliphatic
Carboxylic acid R²COOH〔However, R²is 1 to 21 charcoal
Contains elementary atoms, optionally methoxy, benzyloxy
halogen (e.g. fluorine) or aryl
substituted with compatible substituents such as
Thus, this aryl group has up to 10 carbon atoms.
and optionally lower alkyl, lower alkoxy or
substituted with a halogen group]
includes. Therefore, this acyl group R²CO- is
Formyl (R²=H, using formic anhydride-acetic acid
), acetyl, trifluoroacetyl,
Methoxyacetyl, butyryl, isobutyryl, butyryl
Leryl, octanoyl, lauroyl, myristoy
palmitoyl, behenoyl, phenylacetate
benzoyl, P-fluorobenzoyl, P-
Methoxybenzoyl, naphthoyl, 7-methoxy
Includes groups such as naphthoyl and the like. This selective
The exact conditions for N-acylation depend on the acid used.
Depends on the reactivity of the anhydride. Generally, this reaction
In methanol at about 20° to about 65°C for about 15 minutes.
The reaction proceeded satisfactorily within about 6 hours, and this reaction
Most are also subjected to reflux for about 30 to about 60 minutes.
can become. Certain types of anhydrous trifluoro vinegar
In the case of highly reactive acid anhydrides such as acids,
This reaction is carried out using an inert solution such as tetrahydrofuran.
Performed using a slight excess of acid anhydride in the medium
It is preferable. The compound of formula (8B) in the above reaction is then treated as described above.
Special sodium α-halocarboxylate or
Methyl α-halocarboxylate and sodium
using aluminum hydride in dimethylformamide.
Alkylated, R⁷=H and R²CO−,
R⁶is as defined above.
Zyl 2-acylamido-4,6-o-benzylide
-3-o-(1-carboxyalkyl)-α-D
- produces glucopyranosides. These derivatives
Coupling with the free base derived from formula (10)
is a compound of formula (9) (R²＝CH₃) regarding the generation of
achieved by the same method described and R²CO-acyl
Group, X¹aminoacyl group, n and R⁶is the above definition
The blocked glycope of equation (11) is as follows.
Generate petide. (11) from the blocked glycopeptide of formula
Removal of protecting groups uses a suitable palladium catalyst
Achieved by catalytic hydrolysis. This reaction is usually
5-10% palladium on charcoal, barium sulfate
Palladium, palladium black or related catalysts
1 to 2 atm of hydrogen at a temperature close to room temperature using
Carry out in acidic medium under pressure. Preferably about 60
Palladium on charcoal catalyst in to about 90% aqueous acetic acid
use. Removal of all protecting groups under these conditions
The removal is completed within about 24 to about 72 hours. This exclusion
The removal reaction can be carried out by thin film chromatography if desired.
can be traced, thus formula (13) (R=R¹=
H,R⁶and R²CO− is as defined above
However, R of the compound of formula (11)²is benzyloxymethyl
In this case, this benzyl ether is simultaneously solved.
Split R²A compound of formula (11) where is hydroxymethyl
and X is glycyl, L-alani
L-valyl, L-leucyl, L-isoleucil
L, L-α-aminobutyryl, L-seryl, L-
Threonyl, L-methionyl, L-cysteine
L, L-phenylalanyl, L-tyrosyl, L-
Tryptophanyl, L-lysyl, L-ornichi
L-, L-arginyl, L-histidyl, L-glue
Tamir, L-glutaminyl, L-aspartyl,
L-asparaginyl, L-prolyl or L-hyperginyl
The glycodipeptide raw material of droxyprolyl
product [e.g. 2-(2-acetamido-2-deo
xy-D-glucose-3-O-yl)-D-p
Ropionyl-L-valyl-D-isoglutamine]
generate. These compounds are biorex
(BIOREX) 70 (product name) (weakly acidic polyacrylic acid
Conventional applications such as chromatography on resin) columns
Purify by chromatographic method. depending on the case
Riko's chromatography alone detects small amounts of impurities.
Not completely removed. In such cases, the dow
Dowex 50 (H⁺) (Product name) (Gibi
Sulfonated polyester cross-linked with nylbenzene resin
Effect basic impurities through a styrene column
or amber light
(Amberlite) XAD-2 (trade name) (2% divinyl
Polystyrene cross-linked with benzene) is mixed with water or
is a chroma on a column placed in aqueous methanol.
All partially deblocked by
Separate the glycopeptides. Another aspect of the present invention is glycodype of formula (13)
4,6-di-o-acyl derivatives of peptide (however, R
and R¹are the same or different acyl groups,
X, R²,R⁶and n are as defined above)
Concerning the synthesis of Preferred acyl groups have 1 to 21 carbon atoms.
Containing straight or branched alkyl, aralkyl
aryl, arykaryl or aryl group, e.g.
Cetyl, octanoyl, stearoyl, behenoy
and benzoyl. Furthermore, according to the invention R or R¹one side
is an acyl group as defined above, and the other (R or
R¹) is hydrogen, and X, R²,R⁶and n is
4-o- and 6- of formula (13) as defined
A selective synthesis method for o-acyl derivatives has been developed.
Ta. In the synthesis of all three compounds, their first step
As, the protected glycopeptide of formula (11) (however,
ShiX¹,R²,R⁶and n are as defined above.
selection of 4,6-o-benzylidene functional group from
Using selective acidic hydrolysis. This hydrolysis
Various acid treatments well known in chemical chemistry techniques
Do this using These treatments are approximately 50°.
50-90% acetic acid, formic acid or propylene at about 100℃
Treatment with onic acid or approximately 90% truffle at room temperature
This includes short-term treatments using strong acids such as fluoroacetic acid.
Contains. Advantageously, this reaction is carried out at about 100°C for about 50 minutes.
For about 8 minutes, use about 60% to about 80% acetic acid.
Let's do it. Next, perform the usual finishing treatments and other
4 of equation (12A), which is sufficiently protected in terms of
Isolate the 6-diol and crystallize it or as silicic acid.
and purified by chromatography on a suitable adsorbent.
make (12A) Both free hydroxyls of formula
The conversion is carried out using a suitable inert organic solvent in the presence of a tertiary base.
Among them, acyl anhydride, acyl chloride, acyl
Conventions such as anides or acylimidazolides
2 molar equivalents or
or higher.
Suitable solvents include pyridine, methylene chloride,
Acetonitrile, dimethylformamide, tetra
Including hydrofuran, dioxane, etc., while pyri
gin, triethylamine, N-methylmorpholy
A suitable base such as chlorine, diisopropylethylamine, etc.
It is. 2 to 10 molar equivalents of acyl chloride or
Either of the acylic anhydrides in pyridine at room temperature is approx.
It is generally advantageous to use it for 10 to about 24 hours.
Yes, generates R^Fourand R^Fiveare R and R¹in the case of
is the same acyl group as similarly defined in
Isolating a 4,6-o-acyl derivative of formula (12E),
Crystallization or chromatography on adsorbents such as silicic acid
Purify by graphing. Is it a compound of formula (12E)?
Removal of these protecting groups converts the compound of formula (11) into formula (13).
compound (however, R and R¹is hydrogen)
A palladium-derived catalyst as described above for
This is accomplished by using catalytic hydrocracking. Living
The product formed is R²also encompasses hydroxymethyl
R except that it contains²,R⁶, X and n are as defined above
and R and R¹are the same acyl group
This is a compound of formula (13). Alternatively, a slight excess (approximately 1.0 to 1.2 moles) may be used.
(equivalent) of acyl anhydride, acyl chloride
acyl cyanide or acyl imidazoline
Inert organic solvent containing a tertiary base as described above
Acylation of the compound of formula (12A) used in
results in selective acylation of primary 6-hydroxyl groups
Ru. This reaction occurs at approximately 0° in a solution of the compound of formula (12A).
Add the acylating agent gradually at a temperature of about 20°C.
It is preferable to carry out this reaction with the starting material.
thin film chromatography until substantially consumed.
tracked by Next, finish using the usual method, formula (12F)
(R in the formula³are R and R¹A defined in the same way as
is a syl group, and¹,n,R²and R⁶is the above definition
The 6-o-acyl derivative of
and chromatography on a suitable adsorbent such as silicic acid.
Purify by roughy or crystallization. (12F) formula
Catalytic hydrogenolysis of the compound derived from palladium
Using a catalyst as described above, formula (13) is obtained.
(In the formula R and R²CO− is an ace as defined above.
Syl group, but excludes benzyloxyacetyl
Hydroxyacetyl (glycolyl) group instead
is also added, and R¹is hydrogen)
get something To selectively introduce an acyl group into the 4-o-position
is C in equation (12A).₆Protecting group first on hydroxy group
It is necessary to introduce them selectively. this is a carbohydrate
Using well-known techniques in the chemical field, tritium
or substituted trityl ethers.
It is possible to achieve better binding. In other words,
Dimethylform containing lysine or pyridine
Amide, dimethyl sulfoxide, acetonitrile
The compound of formula (12A) in an inert solvent such as
Triphenylmethyl chloride (trityl chloride)
or anisyldiphenylmethylchloride
(methoxytrityl chloride), dianisilph
enylmethyl chloride (dimethoxytrityl chloride)
chloride) or trianisylmethyl chloride (trianisylmethyl chloride)
methoxytrityl chloride) [this
Regarding the use of reagents, M. Smith et al.
(M.Smith et al)
See J.Amer.Chem.Soc., 84, 430 (1962).
]. Generally, this reaction takes place between about 10° and about 50°C.
It can be carried out in pyridine at room temperature.
is advantageous. The length of time for this reaction is
Depending on the reactant used, trianisylmethyl chloride
changes in about 2 to about 5 hours, or
In the case of triphenylmethyl chloride, it is about 3
It changes in about 6 days. 6 of equation (12B) to generate
-o-trityl or substituted 6-o-trityl ether
The solution may be precipitated, crystallized or
Conventional techniques such as chromatography on adsorbents
It can be easily isolated by surgery. In equation (12B),
R⁶,X¹, n and R²CO− is as defined above.
be. Acyl of the free 4-hydroxyl group of formula (12B)
is an acyl group R³is as defined above
acyl chloride, acyl anhydride,
Acyl cyanide or acyl imidazolide
This is carried out using conventional acylating agents. this reaction
optionally ranges from about 1 to about 5 molar ratios of the acylating agent.
The compound of formula (12A) is
When acylating to produce a compound of formula (12E)
can be carried out under the same conditions as mentioned above.
can. Formula (12C) to generate (where R²,R³,R⁶，
X¹and n are as defined above).
Isolate the silica product and crystallize, precipitate or
by chromatography on a suitable adsorbent such as
Refining. Selective addition of trityl (or substituted trityl) groups
Water splitting can occur under certain acidic conditions.
Wear. This is a clearing solution made with 70% acetic acid at 100℃.
Process until formed, and then process for about 2 minutes more.
It is advantageous to do this by these conditions
should avoid extreme conditions other than temperature, time
Variations can be made by a person skilled in the art with regard to temperature and acidity.
The methoxytrityl derivative corresponding to formula (12C) is
If used, the conditions for this acid hydrolysis are very
It is gentle and can be kept at room temperature for about 5 minutes to about 5 o'clock.
The use of 80% acetic acid is sufficient in between. generate
(12D) Formula (where X¹,n,R⁶,R³and R²is mentioned above
4-o-acyl derivative of
Isolate and crystallize or chromatograph on silicic acid
Purify by b. The 6-hydroxy group of the compound of formula (12D) is
To acylate, use formula (12E) or formula (12C)
Under the conditions outlined for the preparation of the compound of
Bonic acid R^FourUsing an acylating agent derived from OH
and achieve it. Formula (12E) (in the formula, the acyl group
R^Fourand R^Fiveare the same or different as defined above
and R²,R⁶,X¹and n as defined above.
The product is isolated and purified by conventional techniques.
Can be purified. Alternatively, an acid R as described above may be used.^Fivederived from OH
A compound of formula (12F) is prepared using an acylating agent.
By further acylation, the compound of formula (12E)
Manufacture things. Again, the formula (12E) that generates (in the formula
Acyl group R^Fourand R^Fiveare the same or different
As is righteousness, R²,R⁶,X¹and n
is as defined above),
Purify by conventional means. Compounds of formulas (12D), (12E) and (12F)
Removal of the protecting group from formula (13) of the compound of formula (12A)
As described above for the conversion of the acid into the compound
Contact using palladium-derived catalysts in a neutral medium
Achieved by hydrocracking. product produced
is the formula (13) (where R⁶,X,n and R²CO− is in front
as defined above, and R and R¹is the same
one or different acyl groups, or R or
R¹one of which is hydrogen and the other is an acyl group
It is a compound that is either one of the following. either
Also in the case, the acyl group R, R¹,R⁶and R²CO- is
Hydroxya instead of benzyloxyacetyl group
As defined above except for the addition of cetyl (glycolyl) group
It is as follows. These compounds of formula (13) are acetic acid
Less polar solvents such as ethyl or ether
by precipitation from methanol by addition of
and then convert the compound of formula (12A) into formula (13).
As mentioned above regarding the conversion to
on Tux 70 or Amberlight XAD-2
Further purification by chromatography. Apparent
Instead of this, peptides and glycopeptides
Other well-known chromatographs
Roughy technology can be used for these purifications. Isolation of the compounds described herein is optional.
e.g. extraction, crystallization, thin layer chromatography.
B.Thick layer chromatography or column chromatography
Any suitable separation or purification such as graphite
technology or also a combination of these methods.
can become. of suitable separation and isolation methods.
As specific examples, the following examples can be cited. deer
However, other equivalent separation or isolation methods may also be used.
Of course, it can also be used. The term room temperature refers to a temperature of approximately 15° to approximately 25°C.
Taste. The invention may be further understood from the following non-limiting examples.
I can do it. Examples 1 to 14 follow the reaction route described above.
Typical examples related to the production of raw materials and intermediate compounds
It is. Example 1 Carefully add 50 ml of concentrated sulfuric acid to 500 ml of anhydrous ether.
Well, add 500ml of benzyl alcohol to this mixture.
Add. Remove the ether by vacuum evaporation and remove the remaining
The distillate was treated with D-aspartic acid (1,
n=1) Add 67g in portions. The resulting suspension
Stir at 22°C for 16 hours to produce a clear solution.
Add one part of 95% ethanol to this vigorously stirred solution.
and 250 ml of pyridine. Crystallizes after a few minutes
begins and the mixture is kept at 0° C. for 20 hours.
Collect the product by filtration and wash thoroughly with ether.
77.9 g of a white solid with a melting point of 172° to 177°C
get. Mix this product with 1 water containing 10 drops of pyridine.
Recrystallize from the melting point of 220° to 221°C and [α]^{twenty five} _D
-28.1° (10 mg/ml, 1N hydrochloric acid)
Obtained 44.3g of Le D-aspartate (2, n=1)
Ru. In a similar manner, instead of D-aspartic acid, D
- Stoichiometric equivalent of glutamic acid (1, n=2)
using a melting point of 161° to 162°C and [α]^{twenty five} _D−
γ-benzyl D- with 19.3° (6 mg/ml acetic acid)
56.3 g of glutamate (2, n=2) are obtained. Example 2 Anhydrous dimethyl sulfoxy in a 500ml round bottom flask
250 ml of β-benzyl D-aspartate (2,
n=1) 11.15g, triethylamine 13.5ml and
Add 9 ml of t-butyl azide formate and generate
Keep the solution at 22 °C for 42 h. reaction mixture
Then pour the mixture into 700ml of water and 200ml of ether.
Extract 3 times. The aqueous phase was cooled with ice and diluted with acetic acid.
Add 200ml of chill and then quench the PH of this aqueous phase.
Addition of acid brings it to 2.8. Separate the phases and remove the aqueous phase
was further extracted three times with 200 ml of ethyl acetate. gather
The organic phase was mixed with 200 ml of water 5 times and 200 ml of saturated saline.
Extracted continuously with and dried over magnesium sulfate.
let Filter the ethyl acetate solution and evaporate the filtrate to dryness.
The solid residue obtained was dissolved in ethyl acetate-hexane.
crystallized from 102-103°C and
[α]^{twenty five} _D+19.4 (14mg/ml, dimethylformamide)
β-benzyl t-butyloxycarbonyl having
13.2g of Ru-D-aspartate (3 n=1)
obtain. Elemental analysis: C₁₆H_{twenty one}NO.₆For (323.34), Calculated value: C, 59.43; H, 6.55; N, 4.33 Actual values: C, 59.51; H, 6.39; N, 4.28. In a similar manner β-benzyl D-aspartate
stoichiometric equivalent of γ-benzyl D-glue instead of
Using tamate (2, n=2), light yellow oil and
and γ-benzyl t-butyloxycarbonyl-
16.9 g of D-glutamate (3, n=2) are obtained.
This product is pure enough to be used directly in the next step.
have a degree. Example 3 β-benzyl t- in 20 ml of tetrahydrofuran
Butyloxycarbonyl-D-aspartate
(3, n=1) 3.23g and triethyleneamine
Add 1.6 ml of the solution, stir magnetically, and cool to -15℃.
let Add isobutyl chloroformate to this solution.
Add 5 ml of 1.55 ml of tetrahydrofuran solution dropwise.
and maintain temperature at −15° C. for 25 minutes. this reaction
Dry the mixture while maintaining the temperature at -15°C for 30 min.
Bubble dry ammonia gas through. of the reaction mixture
The temperature was then increased to 0°C for 15 minutes;
At this point, the flow of ammonia gas will be stopped. Tetrahi
Evaporate Dorofuran under reduced pressure and dissolve the residue in 80ml of water and vinegar.
and 200 ml of ethyl acid. Ethyl acetate
The phase was washed twice with 50 ml of 5% aqueous sodium bicarbonate, then
Washed twice successively with 50 ml of saturated saline, then rinsed with sulfuric acid.
Dry over magnesium acid. Evaporate the filtered solution
After drying, 3.22 g of the obtained solid was added to 50 ml of ethyl acetate.
Recrystallized from
^{twenty five} _D+17.0 (10mg/ml, dimethylformamide)
benzyl t-butyloxycarbonyl-D
-2.72 g of isoasparaginate (4, n=1)
obtain. Elemental analysis: C₁₆H_{twenty two}N₂O_FiveFor (322.35), Calculated value: C, 59.61; H, 6.88; N, 8.69 Actual value: C, 59.81; H, 6.80; N, 8.87 In a similar manner, β-benzyl t-butyloxy
Instead of carbonyl-D-aspartate, chemical
stoichiometric equivalent of γ-benzyl t-butyloxycarp
Using Bonyl-D-Gludamate (3, n=2)
and melting point of 125° to 126°C; [α]^{twenty two} _D−2.6° (13.3mg
/
benzyl t with ml, dimethylformamide)
-Butyloxycarbonyl-D-isoglutamine
Obtain 1.92 g of salt (4, n=2). Example 4 A benzyl t-butyloxycarbonyl-D-
2.95g of isoglutaminate (4, n=2) and vinegar
Add ethyl acid to 14 ml of a saturated solution of hydrogen chloride. anti
The reaction mixture was kept at 22°C for 10 minutes and then diluted with ether.
Add 80 ml to precipitate the product and discard this precipitate at a distance.
Harvested by heart isolation, carefully dried under reduced pressure,
γ-Benzyl D-isoglutamate hydrochloride
(5, n=2) 2.33g is obtained. This product
Used in Step B of the following example without further purification. B t dissolved in 10 ml of dry dimethylformamide
-Butyloxycarbonyl-L-alanine
(6,X¹= L-alanyl) 2.01g of 1-hydro
1.72g xybenzotriazole and dicyclo
Add 2.19 g of hexylcarbodiimide. Generate
The mixture was kept at 22 °C for 1.5 h and then filtered.
γ-benzyl D-isoglutamate hydrochloride
2.33 g of salt (5, n=2) and diisopropyl
Ethylamine 1.55ml dimethylformamide 10
ml solution. The insoluble obtained from this filtration
Wash the sex cake with 5 ml of dimethylformamide,
Washing solution was added to the reaction mixture and incubated at 22°C for 1 hour.
Hold. Pour the reaction mixture into 500 ml of water,
Extract the resulting solution twice with 250 ml of ethyl acetate.
Ru. The combined organic phase was diluted with bicarbonate of water four times with 200 ml of water.
3 times with 150ml of 5% aqueous solution and saturated salt
Wash continuously with 150 ml of water. This ethyl acetate
The aqueous solution was then dried over sodium sulfate and filtered.
Filtration and evaporation to dryness gave a solid product, which was
crystallized from a mixture of ethanol and ether
melting point of 140° to 141°C and [α]^{twenty two} _D0° (7.6
mg/ml, dimethylformamide)
dil-t-butyloxycarbonyl-L-arani
RuD-isoglutaminate (7, n=2,
X¹= L-alanyl) 2.2 g are obtained. Elemental analysis: C₂₀H₂₉N₃O₆For (407.45), Calculated value: C, 58.95; H, 7.17; N, 10.31 Actual value: C, 59.11; H, 6.80; N, 10.50 In the same way, t-butyloxycarbonyl-L
-Stoichiometric equivalents instead of alanine t-butyloxycarbonyl-L-valine, t-Butyloxycarbonyl-L-α-amino
butyric acid, t-Butyloxycarbonyl-O-benzyl-
L-threonine, t-Butyloxycarbonyl-L-phenyla
Lanin, t-Butyloxycarbonyl-O-benzyl-
L-tyrosine, t-Butyloxycarbonyl-ε-carboben
Zyloxy-L-lysine, t-Butyloxycarbonyl-L-proline
call t-Butyloxycarbonyl-O-benzyl-
L-hydroxyproline and the corresponding protected dipeptide derivative
Obtain a conductor; benzyl t-butyloxycarbonyl-L-ba
Lyle-D-isoglutaminate, melting point: 152°~
153℃; [α]^{twenty five} _D+9.1° (10mg/ml, dimethylform
amide); Elemental analysis: C_{twenty two}H₃₃N₃O₆For (435.50), Calculated value: C, 60.67; H, 7.64; N, 9.65 Actual value: C, 60.88; H, 7.90; N, 9.67 Benzyl t-butyloxycarbonyl-L-α
-aminobutyryl-D-isoglutaminate, fused
Point: 102° ~ 104℃; [α]^{twenty five} _D+3.1° (10mg/ml, moisture
chillformamide); Elemental analysis: C_{twenty one}H₃₁N₃O₆For (421.50), Calculated value: C, 59.84; H, 7.41; N, 9.97 Actual value: C, 59.60; H, 7.56; N, 10.00 benzyl t-butyloxycarbonyl-o-be
Njiru-L-threonyl-D-isoglutamine
Melting point: 117° to 120°C; [α]^{twenty five} _D+7.1° (5.0mg/m
l,
dimethylformamide); Elemental analysis: C₂₈H₃₇N₃O₇For (527.63), Calculated value: C, 63.74; H, 7.07; N, 7.96 Actual value: C, 63.59; H, 6.81; N, 7.84 benzyl t-butyloxycarbonyl-L-ph
enylalanyl-D-isoglutamate, fused
Point: 139° ~ 140℃; [α]^{twenty five} _D−10.7° (10mg/ml, di
Mail
chillformamide); Elemental analysis: C₂₆H₃₃N₃O₆(483.57) for Calculated value: C, 64.58; H, 6.88; N, 8.69 Actual value: C, 64.62; H, 7.17; N, 8.68 benzyl t-butyloxycarbonyl-O-be
undil-L-tyrosyl-D-isoglutamine
Melting point: 172° to 173°C; [α]^{twenty four} _D+4.2° (4.6mg/m
l,
dimethylformamide); Elemental analysis: C₃₃H₃₉N₃O₇for (589.66) Calculated value: C, 67.21; H, 6.67; N, 7.13 Actual value: C, 67.44; H, 6.88; N, 7.06 benzyl t-butyloxycarbonyl-ε-ka
Lebobenzyloxy-L-lysyl-D-isoglu
Taminate, melting point: 94° ~ 97°C; [α]^{twenty five} _D-3.5° (8
mg/ml, dimethylformamide); Elemental analysis: C₃₁H₄₂N_FourO₈(598.68) for Calculated value: C, 62.19; H, 7.07; N, 9.36 Actual value: C, 62.19; H, 7.03; N, 9.44 benzyl t-butyloxycarbonyl-L-propylene
Loryl-D-isoglutaminate, [α]^{twenty four} _D−9.0°
(14.4mg/ml, dimethylformamide)
colorless oil; Elemental analysis: C_{twenty two}H₃₁N₃O₆(433.49) for Calculated value: C, 69.95; H, 7.21; N, 9.69 Actual value: C, 61.06; H, 7.49; N, 9.85 and benzyl t-butyloxycarbonyl-
O-benzyl-L-hydroxyprolyl-D-i
Soglutaminate, [α]^{twenty five} _D−2.4° (5 mg/ml,
oil with chloroformamide). Example 5 A benzyl 2-amino-4,6-o-benzyly
Den-2-deoxy-α-glucopyranoside
(8A) 3.57 g and 2.49 g of benzoic anhydride meth
Heat the 200 ml solution of Nord under reflux for 45 minutes.
The resulting mixture was cooled to 22°C to form a crystalline solid.
Filter, wash with methanol, then add hot methanol
benzyl 2-benzamide by recrystallization from
-4,6-O-benzylidene-2-deoxy-
α-D-glucopyranoside (8B,R²= Ph)
Obtain 3.46g, melting point 218°~220°C; [α]^{twenty five} _D+119°
(10mg/ml, pyridine); Elemental analysis: C₂₇H₂₇NO.₆・H₂for O(479.54)
Ru Calculated value: C, 67.63; H, 6.10; N, 2.92 Actual value: C, 68.09; H, 5.81; N, 2.87 B Benzyl 2-amino-4,6-O-benzyly
Den-2-deoxy-α-D-glucopyranosyl
(8A) 3.57g and trifluoroacetic anhydride
1 hour solution of 2.31g in 100ml of tetrahydrofuran
Heat to reflux for a while. Cool the resulting solution to approximately room temperature.
Allow to cool, then add to ice water 1. precipitated solids
Filter it, wash it thoroughly with water, dry it under reduced pressure, and
Benzyl-4,6-O-benzylidene-2-deo
xy-2-trifluoroacetamide-α-D
- Obtain 4.51 g of glucopyranoside, melting point 242 ~
243℃; [α]^{twenty five} _D+115.2° (9mg/ml, dimethylphosate)
Lumamide); Elemental analysis: C_{twenty two}H_{twenty two}NO.₆F₃・H₂For O(471.44)
do Calculated value: C, 56.05; H, 5.13; N, 2.97 Actual value: C, 56.43; H, 5.18; N, 3.07 C benzyl 2-amino-4,6-o-benzyly
Den-2-deoxy-α-D-glucopyranosyl
(8A) and 2.7 g of methoxyacetic anhydride.
Heat 200 ml of methanol solution to reflux for 30 minutes.
The resulting solution was then concentrated under reduced pressure and the residue was evaporated.
Mix with 150 ml of water, filter, and ventilate.
-4,6-o-benzylidene-2-deoxy
-2-methoxyacetamide-α-D-gluco
Pyranoside (8B, R²=-CH₂OCH₃)5.99g
Obtain, melting point 198° ~ 199°C; [α]^{twenty five} _D+82.0° (10mg/
ml, chloroform); Elemental analysis: C_{twenty three}H₂₇NO.₇(429.47) for Calculated value: C, 64.32; H, 6.34; N, 3.26 Actual value: C, 64.19; H, 6.28; N, 3.06 In a similar manner, instead of benzoic anhydride,
The acylating agent is used in stoichiometric equivalents and the product
using process B or C modified according to the physical properties of
and the corresponding 2-acylamine of the following formula (8B)
Obtain the noglucose derivative: benzyl 2-formamide-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid Benzyl 2-propionamide-4,6-o-
Benzylidene-2-deoxy-α-D-glucopi
Ranocide Benzyl 2-butylamide-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid Benzyl 2-valeramido-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid Benzyl 2-caproamido-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid, melting point 202°~204°C; [α]^{twenty five} _D+108.2° (10mg/
ml, dimethylformamide); Elemental analysis: C₂₆H₃₃NO.₆・0.5H₂For O(464.57)
do Calculated value: C, 76.22; H, 7.38; N, 3.02 Actual value: C, 67.54; H, 7.36; N, 3.10 benzyl 2-heptanamide-4,6-o-be
ndylidene-2-deoxy-α-D-glucopyra
noside Benzyl 2-caprylamido-4,6-o-be
ndylidene-2-deoxy-α-D-glucopyra
noside Benzyl 2-nonanamido-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid benzyl 2-capramide-4,6-o-benzi
Lyden-2-deoxy-α-D-glucopyranosyl
de Benzyl 2-lauramide-4,6-o-benzi
Lyden-2-deoxy-α-D-glucopyranosyl
de Benzyl 2-myristamide-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid, melting point 182° to 184°C; [α]^{twenty five} _D+55.5° (5mg/
ml, chloroform); Elemental analysis: C₃₄H₄₉NO.₆・0.5H₂For O(576.78)
do Calculated value: C, 70.80; H, 8.74; N, 2.43 Actual value: C, 70.50; H, 8.61; N, 2.53 Benzyl 2-palmitamide-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid Benzyl 2-stearamide-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid Benzyl 2-arachidamide-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid Benzyl 2-behenamido-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid Benzyl 2-benzyloxyacetamide-
4,6-o-benzylidene-2-deoxy-α-
D-glucopyranoside, melting point 168° to 169°C; [α]
^{twenty five} _D+72.1° (5 mg/ml, chloroform); Elemental analysis: C₂₇H₃₁NO.₇(505.55) for Calculated value: C, 68.89; H, 6.18; N, 2.77 Actual value: C, 68.82; H, 6.03; N, 2.90 and Benzyl 2-phenylacetamide-4,6-
o-Benzylidene-2-deoxy-α-D-glue
Copyranoside. Example 6 Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid (8) 41.7g dry dimethylformamide 500ml
Add a 50% suspension of sodium hydride in oil to the solution.
Add 1 part at a time to 19.2 g of the suspension. the resulting mixture
Stir for 30 min at 22 °C, then add sodium chloroacetate
Add 23.3g and heat the reaction mixture at 75°C for 1 hour.
Ru. The reaction mixture was cooled in an ice-water bath and diluted with ethanol.
Drop 125ml of the liquid until the foaming subsides (about 30 minutes).
and evaporate the resulting solution to dryness under reduced pressure.
Ru. The residue thus obtained was diluted with chloroform.
ml and a mixture of 500 ml of water and the sodium salt of the product
Stir mechanically until crystallization occurs. Filter the crystals
and resuspended in chloroform 3, then dosed.
Vets 50 (H⁺) (crosslinked with divinylbenzene)
sulfonated polystyrene resin) approx. 450
ml and the mixture was stirred for 1 hour at 22°C. resin
was filtered off, and the chloroform filtrate was diluted with magnesium sulfate.
Dry on top, refilter and then evaporate to dryness under reduced pressure.
benzyl 2-acetamide-4,6-o-
benzylidene-2-deoxy-3-o-carboxy
Cylmethyl-α-D-glucopyranoside (9,
R²＝CH₃) Obtain 39g, melting point 215°-216°C. In a similar manner, benzyl 2-acetamide-
4,6-o-benzylidene-2-deoxy-α-
In place of D-glucopyranoside, stoichiometric equivalents
Using the various compounds obtained in Example 5, the corresponding
A typical example of obtaining a compound of formula (9) is shown below: Benzyl 2-benzyloxyacetamide-
4,6-o-benzylidene-2-deoxy-3-
o-carboxylmethyl-α-D-glucopyrano
Sid, melting point 172°~173°C; [α]^{twenty five} _D+81.4° (5mg/
ml, chloroform); Elemental analysis: C₃₂H₃₅NO.₇(577.61) for Calculated value: C, 66.06; H, 5.90; N, 2.49 Actual value: C, 66.17; H, 5.96; N, 2.58 Benzyl 2-methoxyacetamide-4,6-
o-benzylidene-2-deoxy-3-o-cal
Boxylmethyl-α-D-glucopyranoside, fused
Point 209-210℃; [α]^{twenty five} _D+88.1° (10mg/ml, methane
); Elemental analysis: C_{twenty five}H₂₉NO.₉(487.52) for Calculated value: C, 61.59; H, 6.00; N, 2.87 Actual value: C, 61.40; H, 6.40; N, 2.77 Benzyl 2-benzamido-4,6-o-ben
Zylidene-2-deoxy-3-o-carboxyl
Methyl-α-D-glucopyranoside, melting point 257°~
258℃; [α]^{twenty five} _D+121.8° (10mg/ml, dimethylform
Muamide); Elemental analysis: C₂₉H₂₉NO.₈(519.56) for Calculated value: C, 67.04; H, 5.63; N, 2.70 Actual value: C, 67.23; H, 5.36; N, 2.73 benzyl 2-trifluoroacetamide-4,
6-o-benzylidene-2-deoxy-3-o-
Carboxylmethyl-α-D-glucopyranoside Benzyl 2-caproamido-4,6-o-ben
Zylidene-2-deoxy-3-o-carboxyl
Methyl-α-D-glucopyranoside, melting point 217~
218℃; [α]^{twenty five} _D+106.1° (5 mg/ml, chloroform
); Elemental analysis: C₂₈H₃₅NO.₈(513.60) for Calculated value: C, 65.48; H, 6.87; N, 2.73 Actual value: C, 65.06; H, 6.74; N, 2.75 and Benzyl 2-myristamide-4,6-o-ben
Zylidene-2-deoxy-3-o-carboxyl
Methyl-α-D-glucopyranoside, melting point 188° ~
190℃. Example 6′ Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Sid (8) 41.7g dry dimethylformamide 500ml
Add 9.6 g of 100% sodium hydride (crystals) to the solution.
Add one part at a time and stir the resulting mixture for 30 minutes at 22°C.
Stir to produce the sodium derivative of (8). on the other hand,
7.0g of 100% sodium hydride was added to D,L-2-C.
lorpropionic acid (or stoichiometric equivalents of D,L)
-2-bromopropionic acid) 26g in dry dimethyl
Be careful with the solution cooled on ice in 200 ml of formamide.
and add. The resulting mixture contains sodium
Add the derivative and stir the reaction mixture at 75°C for 3 hours.
Ru. The reaction mixture was cooled to about 5°C in an ice bath and diluted with water.
Add 100 ml and then dimethysulfate over 2 hours.
Add 100ml of water. The temperature of the reaction mixture was brought to 22 °C,
The mixture was then stirred at 22°C for 16 h and a large excess of water was added.
Inject inside. Filter the formed precipitate and wash with water.
Then, dissolve in 500ml of chloroform and remove the aqueous phase.
Remove. Magnesium sulfate chloroform solution
and then concentrated under reduced pressure to obtain the crude ester.
Le (9, R²=R⁶=R⁷＝CH₃) Obtain 49.6g. child
Chroma on a 1 kg column of prepared silicic acid.
treated with ethyl acetate and chloroform.
Elute with a 1:1 mixture of Collect appropriate fractions,
Evaporate to dryness, recrystallize from methanol, and
2-acetamido-4,6-o-benzylyl
Den-2-deoxy-3-o-(D-1-carbo
methoxyethyl)-α-D-glucopyranoside
(9, R²=R⁶=R⁷＝CH₃) Obtain 26.6g, melting point
212-214℃; [α]^{twenty five} _D+126.8° (10mg/ml, chlorophore
(in the room). Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
(rubomethoxyethyl)-α-D-glucopyranosyl
Do (9, R²=R⁶=R⁷＝CH₃) 26.6g methanol
20g of sodium hydroxide in the suspension in 1
1 solution of water was added and the mixture was heated at 100℃ for 1.5 hours.
Heat and allow to cool to room temperature at this point. The generated solution
Concentrate the solution under reduced pressure to a volume of approximately 500ml, and use this solution as
Treat with 6N hydrochloric acid until the pH is 2.5. Generate
Collect the white solid by filtration, wash thoroughly with water, and remove the
dried under vacuum over a tube and then crystallized from methanol.
benzyl 2-acetamido-4,6-o-benzyl
Zylidene-2-deoxy-3-o-(D-1-
carboxyethyl)-α-D-glucopyranoside
(9, R²=R⁶＝CH₃,R⁷=H) Obtain 21.9g, melt
Point 239°~241℃ In a similar manner, D,L-2-chloropropio
In place of the acid, a stoichiometric equivalent of another D,L-2-halo
Alkanonic acids (having 4 to 6 carbon atoms)
(9) (R⁶has 2 to 4 carbons
is an alkyl having an atom and R⁷=H) compound
Typical examples of obtaining things are as follows: Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
ruboxy-1-propyl)-α-D-glucopyra
noside, melting point 216°-217°C; [α]^{twenty five} _D+126.7° (5
mg/ml, dimethylformamide); Elemental analysis: C₂₆H₃₁NO.₈(485.55) for Calculated value: C, 64.32; H, 6.44; N, 2.88 Actual value: C, 64.56; H, 6.22; N, 2.98 and Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
ruboxy-1-pentyl)-α-D-glucopyra
noside, melting point 232°-233°C; [α]^{twenty five} _D+113.2° (5
mg/ml, dimethylformamide); Elemental analysis: C₂₈H₃₅NO.₈(513.60) for Calculated value: C, 65.48; H, 6.87; N, 2.73 Actual value: C, 65.68; H, 7.02; N, 2.77 Similarly, benzyl 2-acetamide-4,6-
o-Benzylidene-2-deoxy-α-D-glue
Instead of copyranoside, the stoichiometric equivalent of Example 5
Using various compounds obtained, the compound of formula (9) was prepared.
A typical example of this is shown below: Benzyl 2-benzyloxyacetamide-
4,6-o-benzylidene-2-deoxy-3-
o-(D-1-carboxyethyl)-α-D-glue
Copyranoside, melting point 220-221℃; [α]^{twenty five} _D+76.8°
(5mg/ml, chloroform); Elemental analysis: C₃₂H₃₅NO.₉(577.61) for Calculated value: C, 66.54; H, 6.11; N, 2.43 Actual value: C, 66.48; H, 6.37; N, 2.46 Benzyl 2-methoxyacetamide-4,6-
o-Benzylidene-2-deoxy-3-o-(D
-1-carboxyethyl)-α-D-glucopyra
Noside, melting point 209-210℃; [α]^{twenty five} _D+107° (3mg/
ml, methanol); Elemental analysis: C₂₆H₃₁NO.₉(501.55) for Calculated value: C, 62.27; H, 6.23; N, 2.79 Actual value: C, 63.00; H, 6.24; N, 2.78 Benzyl 2-benzamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
ruboxyethyl)-α-D-glucopyranoside,
Melting point 275°~276°C; [α]^{twenty five} _D+114.7° (5 mg/ml, di
methylformamide); Elemental analysis: C₃₀H₃₁NO.₈(533.59) for Calculated value: C, 67.53; H, 5.86; N, 2.63 Actual value: C, 67.36; H, 5.94; N, 2.54 benzyl 2-trifluoroacetamide-4,
6-o-benzylidene-2-deoxy-3-o-
(D-1-carboxyethyl)-α-D-glucopi
Ranocide Benzyl 2-caproamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
ruboxyethyl)-α-D-glucopyranoside,
Melting point 234°~236°C; [α]^{twenty five} _D+117.2° (5 mg/ml, di
methylformamide); Elemental analysis: C₂₉H₃₇NO.₈(527.62) for Calculated value: C, 66.02; H, 7.07; N, 2.65 Actual value: C, 66.01; H, 6.88; N, 2.69 and Benzyl 2-myristamide-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
ruboxyethyl)-α-D-glucopyranoside,
Melting point 117°~178°C; [α]^{twenty five} _D+95.5° (5 mg/ml, di
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chillformamide); Elemental analysis: C₃₇H₅₃NO.₈(639.84) for Calculated value: C, 69.46; H, 8.35; N, 2.19 Actual value: C, 69.38; H, 8.21; N, 2.16 Example 6″ Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-α-D-glucopyrano
Dissolve 4.17 g of cid (8) in 30 ml of dry pyridine and add this
The solution is evaporated to dryness under reduced pressure. repeat this process
and then dry the residue with 75 ml of dimethylformamide.
100% sodium hydride (crystals) dissolved in
Add 0.72 g in portions and heat the resulting mixture at 40°C.
Stir for 1 hour to obtain the sodium derivative (8). child
Then the solution of methyl D,L-2-bromodecanoe
(5.0 g of D,L-2-bromodecanonic acid and
From an ethanolic solution containing 31 mmol of azomethane
) 5.3g in 25ml dimethylformamide
and dropwise into the vigorously stirred solution for 15 minutes.
and add. Stir the resulting mixture at 22 °C for 16 h.
and then concentrated to dryness under reduced pressure. Residue produced
is then distributed between 500 ml of ethyl acetate and 250 ml of water.
Then, the ethyl acetate phase was further washed with 100 ml of water and diluted with sulfuric acid.
Dry over magnesium, then concentrate in vacuo to obtain the crude
Made of ester (9, R²=R⁷= Me, R⁶=n-C₈
H₁₇) Obtain 7.7g. This mixture was mixed with ethyl acetate:
400 of silicic acid prepared in loloform (1:9)
chromatography on a g column and ethyl acetate.
Le:chloroform (1:9) 2 to ethyl acetate
Le:Chloroform (3:7) changes linearly to 2
Elute with a solution. Collect appropriate fractions and evaporate to dryness.
Dry, benzyl 2-acetamide-4,6-o
-Benzylidene-2-deoxy-3-o-(D-
1-carbomethoxy-1-nonyl)-α-D-g
Lucopyranoside (9,R²=R⁷= Me, R⁶=n-C₈
H₁₇) Obtain 2,46g; Melting point: 146° to 148°C; [α]^{twenty five} _D
+89.0° (5 mg/ml, chloroform); Elemental analysis: C₃₃H₄₅NO.₈(583.73) for Calculated value: C, 67.90; H, 7.77; N, 2.40 Actual value: C, 68.29; H, 7.77; N, 2.44 Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
(rubomethoxy-1-nonyl)-α-D-glucopi
Ranosside (9,R²=R⁷= Me, R⁶=n-C₈H₁₇)
1.17g in warm (approx. 60°C) methanol 35ml solution
Add a solution of 0.8 g of sodium hydroxide to 35 ml of water as a raw material.
Drop at such a rate that no precipitate is seen.
Add. The resulting solution was heated to reflux for 25 min, then 22
After cooling to ℃, the pH of the solution was adjusted to 2.5 with 6N hydrochloric acid.
Arrange. Immediately add approximately 250 ml of ice/water to the resulting mixture.
In addition, filter the precipitated white solid and thoroughly dissolve with water.
Washed, then vacuum dried over phosphorous pentoxide and vented.
2-acetamido-4,6-o-benzylidene
-2-deoxy-3-o-(D-1-carboxy
-1-nonyl)-α-D-glucopyranoside (9,
R²= Me, R⁶=n-C₈H₁₇,R⁷=H) 1.13g
Obtain; Melting point: 194° to 196°C; [α]^{twenty five} _D+94.7° (5mg/
ml, dimethylformamide); Elemental analysis: C₃₂H₄₃NO.₈(569.70) for Calculated value: C, 67.47; H, 7.61; N, 2.46 Actual value: C, 67.39; H, 7.62; N, 2.55 In a similar manner, methyl D,L-2-bromoteca
stoichiometric equivalent of another methyl D instead of noate,
L-2-bromoalkanoate (5 to 18
(contains a carbon atom)), formula (9) (R⁶teeth
Alkyl having 5 to 18 carbon atoms
R, R⁷A typical example of obtaining a compound with =H) is as follows.
Shown: Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
ruboxy-1-tridecyl)-α-D-glucopi
Ranoside: melting point 157° to 159°C; [α]^{twenty five} _D+77.2° (5
mg/ml, dimethylformamide) and Benzyl 2-acetamido-4,6-o-ben
Zylidene-2-deoxy-3-o-(D-1-ka)
ruboxy-1-heptadecyl)-α-D-gluco
Pyranoside; melting point 155-158℃; [α]^{twenty five} _D+81.4° (5
mg/ml, dimethylformamide). Similarly, benzyl 2-acetamide-4,6-
o-Benzylidene-2-deoxy-α-D-glue
Instead of copyranoside, the stoichiometric equivalent obtained in Example 5
Using various compounds of formula (9),
get something Example 7 A benzyl t-butyloxycarbonyl-L-
Valyl-D-isoglutaminate (7, n=
2.X′=L-baryl) 0.44g of ethyl acetate salt
Add to 5 ml of a saturated solution of hydrogen hydride at 22°C. 20 minutes
After that, add 40ml of ether and centrifuge the precipitated salt.
collected by separation, carefully dried under reduced pressure,
Benzyl L-valyl-D-isoglutamate
Hydrochloride (10, n=2, X'=L-valyl) was obtained
Ru. B The above benzyl L-valyl-D-isoglutami
nate hydrochloride (10, n=2, X'=L-valyl)
diisopropylene in 5 ml of dimethylformamide solution.
Add 0.17ml of pyruethylamine, then add diiso
Propylcarbodiimide 0.139g, 1-hydro
xybenzotriazole 0,178g and ben
Zyl-2-acetamido-4,6-o-benzyly
Den-2-deoxy-3-o-(D-1-cal
boxyethyl)-α-D-glucopyranoside
(9, R²=R⁶＝CH₃,R⁷=H) dimethylform
Add 5 ml of Muamide solution. reaction mixture produced
Stir the mixture magnetically at 22°C and test the reaction using the Kaiser test.
(Kaiser test) to complete the reaction.
Add reactants up to Reaction mixing upon completion of reaction
Pour the substance into excess water and filter out the solids that form.
Wash thoroughly with water, then twice with ethanol.
dried under vacuum over phosphorus pentoxide, and benzyl 2
-(benzyl 2-acetamide-4,6-o-
Benzylidene-2-deoxy-α-D-gluco
pyranoside-3-o-yl)-D-propioni
Ru-L-valyl-D-isoglutamate
(11, n=2, X=L-baryl, R²=R⁶=
CH₃) Obtain 0.74g, melting point 275-277℃ (min.
solution); [α]^{twenty five} _D+76.0° (10 mg/ml, methanol:
Roloform, 15:85); Elemental analysis: C₄₁H₅₀N_FourO₁₁(794.84) for Calculated value: C, 63.55; H, 6.50; N, 7.23 Actual value: C, 63.45; H, 6.51; N, 7.06 In a similar manner, benzyl t-butyl oxycarbyl
Bonyl-L-valyl-D-isoglutamate
Alternatively, use the various products from Example 4 for stoichiometric equivalents.
and the corresponding next protected glycopeptide inducer.
Get the conductor: benzyl 2-(benzyl 2-acetamide-4,
6-o-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-o-yl)-D-propyl
Onyl-L-α-aminobutyryl-D-isoglu
Taminate, melting point 241-262℃; [α]^{twenty five} _D+95.4° (10
mg/ml, dimethylformamide); Elemental analysis: C₄₁H₅₀N_FourO₁₁(774.88) for Calculated value: C, 63.55; H, 6.50; N, 7.23 Actual value: C, 63.66; H, 6.48; N, 7.23 benzyl 2-(benzyl 2-acetamide-4,
6-o-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-o-yl)-D-propyl
onyl-o-benzyl-L-threonyl-D-y
Soglutaminate, melting point 220° to 225°C; [α]^{twenty five} _D+
76.6° (3 mg/ml, dimethylformamide); Elemental analysis: C₄₈H₅₆N_FourO₁₂(881.01) for Calculated value: C, 65.44; H, 6.41; N, 6.36 Actual value: C, 65.78; H, 6.82; N, 6.68 benzyl 2-(benzyl 2-acetamide-4,
6-o-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-o-yl)-D-propyl
onyl-L-phenylalanyl-D-isogluta
laminate, melting point 273° to 277°C; [α]^{twenty three} _D+98.9° (4.5
mg/ml in dimethylformamide); Elemental analysis: C₄₆H₅₂N_FourO₁₁(836.95) for Calculated value: C, 66.01; H, 6.26; N, 6.69 Actual value: C, 66.29; H, 6.27; N, 6.49 benzyl 2-(benzyl 2-acetamide-4,
6-o-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-O-benzyl-L-tyrosyl-D-iso
Glutamate, melting point 276℃ (decomposition); [α]^{twenty four} _D+
85.4° (2.5 mg/ml, dimethylformamide); Elemental analysis: C₅₃H₅₈N_FourO₁₂(943.08) for Calculated value: C, 67.50; H, 6.20; N, 5.94 Actual value: C, 67.49; H, 6.16; N, 5.77 benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-ε-carbobenzyloxy-L-lysyl
-D-isoglutaminate, melting point 253-259°C (min.
solution); [α]^{twenty five} _D+79.2° (10mg/ml, dimethylform
a
mid); Elemental analysis: C₅₁H₆₁N_FiveO₁₃(952.08) for Calculated value: C, 64.34; H, 6.46; N, 7.36 Actual value: C, 64.07; H, 6.72; N, 7.31 benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-prolyl-D-isoglutamine
g, melting point 179° to 180°C; [α]^{twenty four} _D+98.2° (3mg/ml
,
dimethylformamide); Elemental analysis: C₄₂H₅₀N_FourO₁₁(786.89) for Calculated value: C, 64.11; H, 6.40; N, 7.12 Actual value: C, 64.36; H, 6.29; N, 7.01 benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-O-benzyl-L-hydroxyprolyl
-D-isoglutaminate, melting point 75-79℃; [α]
^{twenty five} _D+93.5° (10 mg/ml, dimethylformamide); Elemental analysis: C₄₉H₅₆N_FourO₁₂(893.02) for Calculated value: C, 65.90; H, 6.32; N, 6.27 Actual value: C, 65.71; H, 6.55; N, 6.17 In a similar manner, benzyl 2-acetamide-
4,6-O-benzylidene-2-deoxy-3-
O-(D-1-carboxyethyl)-α-D-glue
Stoichiometric equivalent of benzyl in place of copyranoside
2-acetamido-4,6-O-benzylidene-
2-deoxy-3-O-carboxymethyl-α-
D-glucopyranoside (9,R²＝CH₃)use
and this was combined with each of the appropriate compounds obtained in Example 4.
By reacting the corresponding next protected
Obtain glycopeptide derivatives: benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-acetyl-
D-L-valyl-D-isoglutaminate, melting point
237-239℃; [α]^{twenty five} _D+80.9° (9mg/ml, dimethyl
formamide); Elemental analysis: C₄₂H₅₂N_FourO₁₁(774.84) for Calculated value: C, 63.55; H, 6.50; N, 6.23 Actual value: C, 63.45; H, 6.51; N, 7.06 Benzyl (benzyl 2-acetamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
-α-aminobutyryl-D-isoglutamine
g, melting point 218-230℃; [α]^{twenty five} _D+81.8° (10mg/ml,
dimethylformamide); Elemental analysis: C₄₀H₄₈N_FourO₁₁for (760.86) Calculated value: C, 63.14; H, 6.36; N, 7.36 Actual value: C, 62.94; H, 6.29; N, 7.31 Benzyl (benzyl 2-acetamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-O
-benzyl-L-threonyl-D-isoglutami
nate, melting point 182-188℃; [α]^{twenty five} _D+76.3° (10mg/
ml, dimethylformamide), Benzyl (benzyl 2-acetamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
-phenylalanyl-D-isoglutamate,
Melting point 229°~231°C; [α]^{twenty five} _D+78.6° (5 mg/ml, di
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chillformamide); Elemental analysis: C₄₅H₅₀N_FourO₁₁(822.93) for Calculated value: C, 65.68; H, 6.12; N, 6.81 Actual value: C, 65.63; H, 6.02; N, 6.62 Benzyl (benzyl 2-acetamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-O
-Benzyl-L-tyrosyl-D-isoglutamine
Melting point: 241° to 243°C; [α]^{twenty three} _D+66.7° (4.2mg/
ml, dimethylformamide); Benzyl (benzyl 2-acetamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-ε
-carbobenzyloxy-L-lysyl-D-iso
Glutaminate, melting point 215° to 219°C; [α]^{twenty five} _D+
66.7° (5.5 mg/ml, dimethylformamide); Elemental analysis: C₅₀H₅₉N_FiveO₁₃(938.06) for Calculated value: C, 64.02; H, 6.34; N, 7.47 Actual value: C, 63.79; H, 6.51; N, 7.44 Benzyl (benzyl 2-acetamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
-Prolyl-D-isoglutaminate, melting point 224°
~225℃; [α]^{twenty four} _D+68.1° (3.1mg/ml, dimethylphosate)
Lumamide); Elemental analysis: C₄₁H₄₈N_FourO₁₁(772.82) for Calculated value: C, 63.72; H, 6.26; N, 7.25 Actual value: C, 63.49; H, 6.39; N, 7.14 Benzyl (benzyl 2-acetamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-O
-benzyl-L-hydroxypurulyl-D-iso
Glutaminate, melting point 219°~221°C; [α]^{twenty five} _D+
73.2° (9 mg/ml, dimethylformamide); Elemental analysis: C₄₈H₅₄N_FourO₁₂(878.99) for Calculated value: C, 65.59; H, 6.19; N, 6.37 Actual value: C, 65.49; H, 6.42; N, 6.33 In a similar manner, benzyl 2-acetamide-
4,6-O-benzylidene-2-deoxy-3-
O-(D-1-carboxyethyl)-α-D-glue
Instead of copyranoside, the compounds obtained in Example 6, 6′ and 6″
Using stoichiometric equivalents of the compounds obtained, these are used as examples.
By reacting with the appropriate compound obtained in step 4, various pairs were prepared.
The following is a representative example of the compound of formula (13) that corresponds to
show: benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
glucopyranoside-3-O-yl)-D-butyryl
Leu-L-valyl-D-isoglutamate, melting point
273°～276℃; [α]^{twenty five} _D+94.3° (1.4 mg/ml, dimethyst
le
formamide); Elemental analysis: C₄₃H₅₄N_FourO₁₁(802.94) Calculated value: C, 64.32; H, 6.78; N, 6.98 Actual value: C, 64.00; H, 6.73; N, 6.81 benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
glucopyranoside-3-O-yl)-D-butyryl
L-L-prolyl-D-isoglutamate,
[α]^{twenty five} _D+87.6° (5 mg/ml, dimethylformamide
)
oil having; Elemental analysis: C₄₃H₅₂N_FourO₁₁(800.92) for Calculated value: C, 64.48; H, 6.57; N, 7.00 Actual value: C, 64.20; H, 6.52; N, 7.19 benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
glucopyranoside-3-O-yl)-D-hexa
Noyl-L-valyl-D-isoglutamate,
Melting point 264°~266°C; [α]^{twenty five} _D+76.6° (3 mg/ml, di
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chillformamide); Elemental analysis: C₄₅H₅₈N_FourO₁₁(830.99) for Calculated value: C, 65.04; H, 7.04; N, 6.74 Actual value: C, 65.23; H, 7.27; N, 6.81 benzyl 2-(benzyl 2-acetamide-4,
6-O-Benzylidene-2-deoxy-α-D-
glucopyranoside-3-O-yl)-D-hexa
Noyl-L-prolyl-D-isoglutamine
g, melting point 80° to 88°C; [α]^{twenty five} _D+85.7° (5mg/ml,
Ji
methylformamide); Elemental analysis: C₄₅H₅₆N_FourO₁₁(828.98) for Calculated value: C, 65.20; H, 6.81; N, 6.76 Actual value: C, 65.43; H, 6.86; N, 6.84 Benzyl (benzyl 2-benzamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
- Valyl-D-isoglutaminate, melting point 242° ~
249℃; [α]^{twenty five} _D+74.4° (5mg/ml, dimethylform
amide); Elemental analysis: C₄₆H₅₂N_FourO₁₁(836.95) for Calculated value: C, 66.01; H, 6.26; N, 6.69 Actual value: C, 65.82; H, 6.20; N, 6.43 benzyl 2-(benzyl 2-benzamide-4,
6-O-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-valyl-D-isoglutamate,
Melting point 257°~259°C; [α]^{twenty five} _D+73.2° (5 mg/ml, di
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chillformamide); Elemental analysis: C₄₇H₅₄N_FourO₁₁for (850.98) Calculated value: C, 66.34; H, 6.40; N, 6.58 Actual value: C, 66.54; H, 6.39; N, 6.50 benzyl 2-(benzyl 2-benzamide-4,
6-O-Benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-prolyl-D-isoglutamine
g, melting point 148° to 151°C; [α]^{twenty five} _D+60.8° (5mg/ml
,
dimethylformamide); Elemental analysis: C₄₇H₅₂N_FourO₁₁(849.97) for Calculated value: C, 66.49; H, 6.17; N, 6.60 Actual value: C, 66.32; H, 6.25; N, 6.58 Benzyl (benzyl 2-caproamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
- Valyl-D-isoglutaminate, melting point 264° ~
265℃; [α]^{twenty five} _D+74.5° (5 mg/ml, dimethylform
amide); Elemental analysis: C₄₅H₅₈N_FourO₁₁(830.99) for Calculated value: C, 65.04; H, 7.04; N, 6.74 Actual value: C, 64.92; H, 6.97; N, 6.74 Benzyl (benzyl 2-caproamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)acetyl-L-
Prolyl-D-isoglutaminate, melting point 193° ~
194℃; [α]^{twenty five} _D+58.0° (5mg/ml, dimethylform
amide); Elemental analysis: C₄₅H₅₆N_FourO₁₁(828.98) for Calculated value: C, 65.20; H, 6.81; N, 6.77 Actual value: C, 65.03; H, 6.61; N, 6.85 benzyl 2-(benzyl 2-caproamide-4,
6-O-benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-valyl-D-isoglutamate,
Melting point 252°~253°C; [α]^{twenty five} _D+86.7° (5 mg/ml, di
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chillformamide); Elemental analysis: C₄₆H₆₀N_FourO₁₁(845.02) for Calculated value: C, 65.38; H, 7.16; N, 6.63 Actual value: C, 65.66; H, 6.99; N, 6.74 benzyl 2-(benzyl 2-caproamide-4,
6-O-benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-prolyl-D-isoglutamine
g, melting point 136° to 138°C; [α]^{twenty five} _D+86.0° (5mg/ml
,
dimethylformamide), Benzyl (benzyl 2-myristamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
- Valyl-D-isoglutamate, Benzyl (benzyl 2-myristamide-4,6
-O-benzylidene-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
-prolyl-D-isoglutaminate, benzyl 2-(benzyl 2-myristamide-4,
6-O-benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-valyl-D-isoglutamate, and benzyl 2-(benzyl 2-myristamide-4,
6-O-benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-prolyl-D-isoglutamine
to. Example 8 Heat 45.0 ml of 67% acetic acid to 97°C on a steam bath, and
Zyl 2-(benzyl 2-acetamide-4,6-
O-benzylidene-2-deoxy-α-D-glue
Copyranosid-3-O-yl)-D-propioni
Ru-L-valyl-D-isoglutamate (11,
n=2,X¹=L-Baryl, R²=R²＝CH₃)5.9g
Add. Generate solution at 97° to 100° for exactly 7 minutes
℃ and then immediately cooled in an ice bath. melt
The liquid is evaporated to dryness under reduced pressure, and the residue is separated from water.
Acetic acid is removed by evaporation and co-evaporation with toluene.
to obtain the crude product. 250 ether to product
ml and the residue is stirred magnetically for 15 h at 22 °C.
The resulting solid was collected by filtration, dried and benzyl 2-
(Benzyl 2-acetamido-2-deoxy-α
-D-glucopyranoside-3-O-yl)-D-
Propionyl-L-valyl-D-isoglutamine
(12A, n=2, X′=L-baryl, R²=R⁶
＝CH₃); melting point 221-224℃; [α]^{twenty five} _D+
103.5° (10mg/ml, dimethylformamide); Elemental analysis: C₃₅H₄₈N_FourO₁₁for (700.80) Calculated value: C, 59.99; H, 6.90; N, 8.00 Actual value: C, 59.56; H, 6.92; N, 7.70 In a similar manner, benzyl 2-(benzyl 2-
Acetamide-4,6-O-benzylidene-2-
Deoxy-α-D-glucopyranoside-3-O-
yl)-D-propionyl-L-valyl-D-y
Compound obtained in Example 7 instead of soglutamate
is used in stoichiometric equivalents, and the corresponding equation (12A) is
Representative examples of obtaining compounds are shown below: Benzyl (benzyl 2-acetamido-2-de
Oxy-α-D-glucopyranoside-3-O-y
)-acetyl-L-valyl-D-isoglutami
nate, melting point 215°~222°C; [α]^{twenty five} _D+79.0° (10mg
/
ml, dimethylformamide); Elemental analysis: C₃₄H₄₆N_FourO₁₁(686.77) for Calculated value: C, 59.46; H, 6.75; N, 8.16 Actual value: C, 59.56; H, 6.94; N, 7.92 Example 9 benzyl 2-(benzyl 2-acetamide-2
-deoxy-α-D-glucopyranoside-3-O
-yl)-D-propionyl-L-valyl-D-
Isoglutaminate (12A, n = 2, X' = L-bar)
Lil, R²=R⁶＝CH₃) 0.07g and triphenylmethy
A solution of 0.56 g of luchloride in 10 ml of pyridine for 5 days.
Stir at 22°C and then steam dry under reduced pressure. residual
Divide the material between 30 ml of chloroform and 30 ml of water, and
Extract the loloform phase two or more times with 30 ml of water, and
Dry over magnesium, wash with ether and dry
to convert benzyl 2-(benzyl 2-acetamide
Do-2-deoxy-6-O-triphenylmethyl
-α-D-glucopyranoside-3-O-yl)-
D-propionyl-L-valyl-D-isogluta
Minate (12B, n=2, X'=L-baryl, R²=
R³＝CH₃). In a similar manner, benzyl 2-(benzyl 2-a
Cetamido-2-deoxy-α-D-glucopyra
Nosido-3-O-yl)-D-propionyl-L
- instead of valyl-D-isoglutamate Example 8
Using the compound obtained in stoichiometric equivalents,
A compound of formula (12B) is obtained. Example 10 benzyl 2-(benzyl 2-acetamide-2
-deoxy-α-D-glucopyranoside-3-O
-yl)-D-propionyl-L-valyl-D-
Isoglutaminate (12A, n=2, X=L-bar)
Lil, R²=R⁶＝CH₃)0.36g and 1.0ml of acetic anhydride.
A 5 ml solution of pyridine was kept at 22°C for 16 hours.
At this point, add 1 ml of methanol and bring the resulting solution to room temperature.
Stir for an additional hour at warm temperature. The solution is then evaporated to dryness
and divide the residue between 20 ml of chloroform and 20 ml of water.
the chloroform phase over magnesium sulfate.
Dry and then evaporate to dryness to obtain the crude product.
This product was crystallized from aqueous ethanol and
benzyl 2-(benzyl 2-acetamide-4,
6-di-O-acetyl-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-valyl-D-isoglutamate
(12E, n=2, X′=L-baryl, R²=R⁶＝CH₃，
R^Four=R^Five＝COCH₃), melting point 225°~241°C;
[α]^{twenty five} _D+93.7° (10mg/ml, dimethylformamide
Do); Elemental analysis: C₃₉H₅₂N_FourO₁₃(748.88) for Calculated value: C, 59.68; H, 6.68; N, 7.14 Actual value: C, 60.00; H, 6.76; N, 7.14 In a similar manner, substitute the appropriate acyl for acetic anhydride.
oxidizing agent (acid anhydride or acid chloride) at stoichiometry.
to obtain the corresponding compound of formula (12E),
Typical examples are shown below: benzyl 2-(benzyl 2-acetamide-4,
6-di-O-butyryl-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-valyl-D-isoglutamate, benzyl 2-(benzyl 2-acetamide-4,
6-di-O-octanoyl-2-deoxy-α-
D-glucopyranoside-3-O-yl)-D-p
Ropionyl-L-valyl-D-isoglutamine
g, melting point 207° to 210°C; [α]^{twenty five} _D+79.3° (8mg/ml
,
dimethylformamide), benzyl 2-(benzyl 2-acetamide-4,
6-di-O-stearoyl-2-deoxy-α-
D-glucopyranoside-3-O-yl)-D-p
ropionyl-L-valyl-isoglutamate,
Melting point 200°~207°C, benzyl 2-(benzyl 2-acetamide-4,
6-di-O-behenoyl-2-deoxy-α-D
-glucopyranoside-3-O-yl)-D-pro
Pionyl-L-valyl-D-isoglutamine
g, melting point 202-205℃; [α]^{twenty five} _D+68.1° (5mg/ml,
dimethylformamide), and benzyl 2-(benzyl 2-acetamide-4,
6-di-O-benzoyl-2-deoxy-α-D
-glucopyranoside-3-O-yl)-D-pro
Pionyl-L-valyl-D-isoglutamine
Melting point: 169° to 172°C. In a similar manner, benzyl 2-(benzyl 2-a
Cetamido-2-deoxy-α-D-glucopyra
Nosido-3-O-yl)-D-propionyl-L
- instead of valyl-D-isoglutamate Example 8
Using stoichiometric equivalents of the compound obtained in
A typical example of the compound of formula (12E) is as follows.
Shown: benzyl 2-(benzyl 2-acetamide-4,
6-di-O-acetyl-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-prolyl-D-isoglutamine
g, melting point 80° to 85°C; [α]^{twenty five} _D+82.4° (10mg/ml,
Ji
methylformamide); Elemental analysis: C₃₉H₅₀N_FourO₁₃for (782.86) Calculated value: C, 59.84; H, 6.44; N, 7.16 Actual value: C, 59.58; H, 6.03; N, 7.13 benzyl 2-(benzyl 2-acetamide-4,
6-di-O-butyryl-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-prolyl-D-isoglutamine
to, benzyl 2-(benzyl 2-acetamide-4,
6-di-O-octanoyl-2-deoxy-α-
D-glucopyranoside-3-O-yl)-D-p
Ropionyl-L-prolyl-D-isoglutamine
tort, Benzyl (benzyl 2-acetamide-4,6
-di-O-acetyl-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
- Valyl-D-isoglutaminate, melting point 197°~
205℃; [α]^{twenty five} _D+79.1° (10mg/ml, dimethylform
amide); Elemental analysis: C₃₈H₅₀N_FourO₁₃for (770.85) Calculated value: C, 59.21; H, 6.54; N, 7.27 Actual value: C, 59.55; H, 6.33; N, 7.28 Benzyl (benzyl 2-acetamide-4,6
-di-O-butyryl-2-deoxy-α-D-g
Lucopyranoside-3-O-yl)-acetyl-L
- Valyl-D-isoglutamate, and Benzyl (benzyl 2-acetamide-4,6
-di-O-octanoyl-2-deoxy-α-D
-glucopyranoside-3-O-yl)-acetyl
-L-valyl-D-isoglutaminate, melting point
174-177℃; [α]^{twenty five} _D+68.3° (8 mg/ml, dimethyl
formamide). Example 11 Benzyl 2 (benzyl 2-acetamido-2-
Deoxy-α-D-glucopyranoside-3-O-
yl)-D-propionyl-L-valyl-D-y
Benzyl 2-(benzi) instead of soglutamate
2-acetamido-2-deoxy-6-O-t
rifenylmethyl-α-D-glucopyranoside-
3-O-yl)-D-propionyl-L-valyl
-D-isoglutaminate (12B, n=2,¹=
L-baryl, R²=R⁶＝CH₃) in stoichiometric equivalents
Repeat the method of Example 10 except that benzyl 2
-(benzyl 2-acetamido-4-O-acetyl
-2-deoxy-6-O-triphenylmethyl
-α-D-glucopyranoside-3-O-yl)-
D-propionyl-L-valyl-D-isogluta
laminate (12C, n=2, X¹=L-Baryl, R²
=R⁶＝CH₃,R³＝COCH₃). In a similar manner, substitute the appropriate acyl for acetic anhydride.
oxidizing agent (either acid anhydride or acid chloride)
Use stoichiometric equivalents and convert the corresponding formula (12C)
Obtain a compound. In a similar manner, benzyl 2-(benzyl 2-a
Cetamido-2-deoxy-6-O-tripheni
methyl-α-D-glucopyranoside-3-O-
yl)-D-propionyl-L-valyl-D-y
Compound obtained in Example 9 instead of soglutamate
is used in stoichiometric equivalents, and the corresponding equation (12C) is
Obtain the compound. Example 12 benzyl 2-(benzyl 2-acetamide-4
-O-acetyl-6-O-triphenylmethyl-
α-D-glucopyranoside-3-O-yl)-D
-Propionyl-L-valyl-D-isoglutami
Nate (12C, n=2, X¹=L-Baryl, R²=
R⁶＝CH₃,R³=-COCH₃) 0.97g and 70% acetic acid 10
ml and heated at 100℃ until a clear solution is obtained.
(about 2 minutes). Place this solution directly in an ice bath.
Immediately cool the mixture and filter off the precipitate. Evaporate the filtrate under reduced pressure.
Let dry and mix the residue with 25 ml of ether.
The resulting solid was collected by filtration, washed with ether, and dried under reduced pressure.
After drying, benzyl-2-(benzyl 2-acetoacetate)
Mido-4-O-acetyl-α-D-glucopyrano
Sid-3-O-yl)-D-propionyl-L-
Valyl-D-isoglutaminate (12C, n=
2,X¹=L-Baryl, R²=R⁶＝CH₃,R³=-
COCH₃) is generated. In a similar manner, benzyl 2-(benzyl 2-a
Cetamide-4-O-acetyl-6-O-trif
enylmethyl-α-D-glucopyranoside-3-
O-yl)-D-propionyl-L-valyl-D
- obtained in Example 11 instead of isoglutaminate
Compounds are used in stoichiometric equivalents and correspond (12D)
A compound of formula is obtained. Example 13 benzyl 2-(benzyl 2-acetamide-2
-deoxy-α-D-glucopyranoside-3-O
-yl)-D-propionyl-L-valyl-D-
Instead of isoglutamate, benzyl 2-(benzyl)
2-acetamido-4-O-acetyl-α
-D-glucopyranoside-3-O-yl)-D-
Propionyl-L-valyl-D-isoglutamine
(12D, n=2,¹=L-Baryl, R²=R⁶
＝CH₃,R³=-COCH₃) in stoichiometric equivalents
and using propionic anhydride instead of acetic anhydride.
Repeat the method of Example 10 except that benzyl 2-
(Benzyl 2-acetamido-4-O-acetyl
-6-O-propionyl-α-D-glucopyrano
Sid-3-O-yl)-D-propionyl-L-
Valyl-D-isoglutaminate (12E, n=
2,X¹=L-Baryl, R²=R⁶＝CH₃,R^Five=
COC₂H₃,R^Four=-COC₂H_Five). In a similar manner, suitable instead of propionic anhydride
acylating agent (acid anhydride or acid chloride)
Used in stoichiometric equivalents, the corresponding compound of formula (12E)
get something In a similar manner, benzyl 2-(benzyl 2-a
Cetamide-4-O-acetyl-α-D-gluco
pyranosid-3-O-yl)-D-propionyl
- instead of L-valyl-D-isoglutamate
Using the compound obtained in Example 12 in stoichiometric equivalents,
The corresponding compound of formula (12E) is obtained. Example 14 benzyl 2-(benzyl 2-acetamide-2
-deoxy-α-D-glucopyranoside-3-O
-yl)-D-propionyl-L-valyl-D-
Isoglutaminate (12A, n=2, X¹=L-
Baril, R²=R⁶＝CH₃)0.36g dissolved in 5ml of pyridine
The solution was mixed with 0.095 ml of acetic anhydride and 1 ml of pyridine for 30 minutes.
Stir at 0° C. while adding dropwise. Living
The resulting solution was kept at 0°C for 16 hours, at which point the solution was
Add 1 ml of tanome and evaporate the mixture to dryness.
The residue thus obtained was mixed with 100 ml of chloroform.
Partition the chloroform phase between 100 ml of water and sulfuric acid macerate.
Dry over magnesium and concentrate under reduced pressure to yield the crude product.
Produces 0.36g of substance. This product is separated by thick layer chromatography.
Benzyl 2-(benzyl 2-(benzyl)
Dyl-2-acetamido-6-O-acetyl-2-
Deoxy-α-D-glucopyranoside-3-O-
yl)-D-propionyl-L-valyl-D-y
Soglutaminate (12F, n=2, X¹=L-Bali
Le, R²=R⁶＝CH₃,R³=-COCH₃) is generated.
Ru. In a similar manner, substitute the appropriate acyl for acetic anhydride.
oxidizing agent (either acid anhydride or acid chloride)
Use stoichiometric equivalents and convert the corresponding formula (12F)
Obtain a compound. In a similar manner, benzyl 2-(benzyl 2-a
Cetamido-2-deoxy-α-D-glucopyra
Nosido-3-O-yl)-D-propionyl-L
- instead of valyl-D-isoglutamate Example 8
Using the compound obtained in stoichiometric equivalents, (12F)
A compound of formula is obtained. Example 15 benzyl 2-(benzyl 2-acetamide-4,
6-O-benzylidene-2-deoxy-α-D-
Glucopyranoside-3-O-yl)-D-propyl
onyl-L-valyl-D-isoglutamate
(11, n=2,¹=L-Baryl, R²=R⁶＝CH₃)
85% of 0.50g and 50mg of 10% palladium on carbon catalyst
The mixture in 15 ml of acetic acid was heated at 22°C in a hydrogen atmosphere.
Stir magnetically. Product (13, n=2, X=L
-Baryl, R²=R⁶＝CH₃) formation is 8:1:1
A solvent consisting of acetonitrile:acetic acid:water in the ratio of
Tracked by thin film chromatography using
Ru. This reaction is generally complete in 24-40 hours
I understand. At this point, use a microporous filter [Teflon
(Product name)] to remove the catalyst and remove the filtrate.
is evaporated to dryness under reduced pressure to obtain a residue. This residual
Dissolve the substance in 3 ml of methanol, then add 8 ml of ethyl acetate.
Precipitate the product by adding ml. thus obtained
The resulting white solid was collected by centrifugation and
Dry carefully to obtain a white powder. This powder
Dissolve in 5 ml of water and add this solution to Biorex.
〓(BIOREX) 70 (Polyacrylic acid weak acid ion
Pass it over a 20 cm x 0.5 cm column of ion-exchange resin).
The column was eluted with water to remove any residue containing the desired product.
The fractions obtained were collected, freeze-dried in vacuum, and
Cetamido-2-deoxy-D-glucose-3
-O-yl)-D-propionyl-L-valyl-
D-isoglutamine (13, n=2, X=L-bari
Le, R=R¹= hydrogen, R²=R⁶＝CH₃) produces 200mg
Melting point: 140° to 158°C (decomposition); [α]^{twenty four} _D+13.6°
(10mg/ml, methanol); Elemental analysis: C_{twenty one}H₃₆N_FourO₁₁(538.57) for Calculated value: C, 46.83; H, 7.11; N, 10.40 Actual value: C, 46.63; H, 7.23; N, 10.26 In a similar manner, benzyl 2-(benzyl 2-a
Cetamido-4,6-O-benzylidene-2-de
Oxy-α-D-glucopyranoside-3-O-y
)-D-propionyl-L-valyl-D-iso
Instead of glutamate, various formations from Example 7
Using stoichiometric equivalents of
Obtain lycopeptide. 2-(2-acetamido-2-deoxy-D-
glucose-3-O-yl)-D-propionyl
-L-phenylalanyl-D-isoglutamine
(Reference compound), melting point 140° to 143°C; [α]^{twenty two} _D+84.4
°
(5.9mg/ml, methanol); Elemental analysis: C_{twenty five}H₃₆N_FourO₁₁・11/2H₂O (595.62) against Calculated value: C, 50.41; H, 6.60; N, 9.41 Actual value: C, 50.25; H, 6.25; N, 9.27 In a similar manner, benzyl 2-(benzyl 2-a
Cetamido-4,6-O-benzylidene-2-de
Oxy-α-D-glucopyranidoside-3-O-
yl)-D-propionyl-L-valyl-D-y
Instead of soglutamate, various raw materials from Example 10
Using the components in stoichiometric equivalents, the corresponding equation (13) is
Obtain the compound. Typical examples of the present invention thus obtained
The following compounds are: 2-(4,6-di-O-acetyl-2-aceto
Amido-2-deoxy-D-glucose-3-O
-yl)-D-propionyl-L-valyl-D-
Isoglutamine; melting point 150° to 170°C (decomposition); [α]
^{twenty five} _D+28.3° (10mg/ml, methanol); Elemental analysis: C_{twenty five}H₄₀N_FourO₁₃・2H₂For O(640.63)
do Calculated value: C, 46.87; H, 6.92; N, 8.75 Actual value: C, 46.93; H, 6.22; N, 8.61 2-(4,6-di-O-octanoyl-2-a
Cetamido-2-deoxy-D-glucose-3
-O-yl)-D-propionyl-L-valyl-
D-isoglutamine; melting point 198° to 200°C; [α]^{twenty five} _D
+45.9° (5 mg/ml, methanol); Elemental analysis: C₃₇H₆₄N_FourO₁₃(772.95) for Calculated value: C, 57.49; H, 8.35; N, 7.25 Actual value: C, 57.52; H, 8.27; N, 7.26 2-(4,6-di-O-stearoyl-2-a
Cetamido-2-deoxy-D-glucose-3
-O-yl)-D-propionyl-L-valyl-
D-isoglutamine; melting point 187℃, 2-4,6-di-O-behenoyl-2-aceto
Amido-2-deoxy-D-glucose-3-O
-yl)-D-propionyl-L-valyl-D-
Isoglutamine; melting point 175° to 185°C, 2-(4,6-di-O-benzoyl-2-acetate)
toamido-2-deoxy-D-glucose-3-
O-yl)-D-propionyl-L-valyl-D
-isoglutamine; melting point 152°C (decomposed), Well known to those familiar with carbohydrate chemistry
By using techniques, especially crystallization
By doing so, equation (13) [and (or) ()
The compound of formula] into individual α- and β-anomers
To divide. Example 16 20μg of tyrosine arsanilate and protein [cattle
Serum albumin (BSA) or ovalbumin
Use an antigen mixture consisting of 100 μg of either
do. This antigen mixture and test compound are shown in the table below.
Dissolved in saline at the indicated dose/guinea pig, etc.
Breast with amount of Freund's incomplete adjuvant (FIA)
to become A group of 10 guinea pigs is listed separately.
Unless otherwise specified, each test compound is used separately, and each guinea pig is used separately.
Inject 0.1ml of the above emulsion subcutaneously into the eel area.
Ru. In all analyses, the control group was emulsified with FIA.
providing said antigen mixture and in most analyses,
The test compound was also 2-(2-acetamido-2-
deoxy-D-glucose-3-O-yl)-D
-Propionyl-L-alanyl-D-isogluta
Compare with Min under the same conditions. Binds to BSA or ovalbumin after 14 days
25 μg of tyrosine arsanilate in 0.1 ml of saline solution
This method is used to inject intradermally into the animal's flank.
skin test on all animals (antibiotics).
This occurs when ovalbumin is present in the raw mixture.
BSA is used in the test, and egg white alcohol is used in the reverse case.
using Bumin). Infiltration observed in each skin test
A number from 1 to 3 is a measure of the size of a 24-hour period.
By multiplying the average area of skin reaction at
Delayed hypersensitivity reaction to tyrosine rusanilate
Calculate the potency of the stimulating test compound. Hidden
The expression is as follows:
The skin test indicated is recorded as ++ and given the value 3;
Those showing a lesser degree of infiltration are marked with a +.
and give a value of 2; on the other hand, a skin reaction that does not show infiltration
The response is represented by 0 and the number 1 is assigned. these numbers
are then summed for each group of animals, and the resulting average value is
Multiply by the average area of observed skin reactions. write
Therefore, the titer includes the average area observed in the skin test.
Combined degrees of infiltration are included. of the test compound
The average titer is shown in the first column of the table below and has a value of 1.
Compare with FIA. All animals were bled by cardiac puncture on day 42;
I¹²⁵FAR using labeled bovine serum albumin
(Farr) technology [P. Minden]
and R.S.Farr,
“Handbook of Experimental I”
Handbook of Experimental
Immunology), D.M.
Weir), 15 chapters, Volume 1, Black Weir
Yenteifutsk Publications, Otsuk
By radioimmunoassay using
using the passive hemagglutination method to convert the serum into an anti-bovine serum aliquot.
minutes for rubumin or anti-ovalbumin antibodies.
analyze. The average titers for these analyzes are shown in column 3 of the following table.
A comparison with FIA, which has a value of 1, is shown below. On day 42, all surviving animals
protein (if BSA is present in the antigen mixture).
BSA in case or ovalbumin in vice versa)
Perform a skin test using 10 μg. 1 in 24 hours
For this protein compared to FIA with numerical values
Potency of test compound to stimulate delayed-type hypersensitivity reactions
, delayed-type hypersensitivity to tyrosine arsanilate
Measured exactly as described above for the reaction.
and is shown in the second column of the table below.

【table】

[Table] Example 17 Acute toxicity test in mice Method: The test substance is dissolved or homogeneously suspended in an aqueous medium containing 0.9% sodium chloride, 0.5% sodium carboxymethylcellulose, 0.4% polysorbate-80 and 0.9% benzyl alcohol. Ta. The concentration was adjusted so that the indicated dose could be administered in a volume of 10ml/1Kg of body weight. The test substance was then subcutaneously injected (S.
C.) and the animals were observed daily for 21 days for mortality. A control group of mice not receiving drug was tested at the same time. As can be seen from the accompanying table, all of the tested compounds showed LD ₅₀ values greater than 60 mg/Kg body weight. This particular dose is approximately the amount required to elicit enhanced delayed-type hypersensitivity and to circulate the antibody levels described in the examples.
I chose it because it is equivalent to 30 times.

【table】

【table】

Claims (1)

[Claims] 1 formula [In the formula, R and R ¹ are the same, and acetyl,
represents octanoyl, stearoyl, behenoyl or benzoyl; R ² represents methyl; R ⁶
represents methyl; X represents L-valyl; Y
represents D-isoglutamine; and the wavy line (〓) represents α- or β-configuration or a mixture thereof, where if one of the wavy lines is α, the other is β. 2. A compound according to claim 1, wherein R and R ¹ are acetyl, octanoyl, stearoyl, behenoyl or benzoyl groups. 3. A compound according to claim 1, wherein R and R ¹ are both acetyl. 4. The compound according to claim 1, wherein R and R ¹ are both octanoyl. The compound according to claim 1, wherein 5 R and R ¹ are both stearoyl. 6 formula; where R and R ¹ are the same and represent acetyl, octanoyl, stearoyl, behenoyl or benzoyl; R ² represents methyl; R ⁶ represents methyl; X is L-valyl; Y is D - isoglutamine; and the wavy line (〓) represents the α- or β-configuration, or a mixture thereof, where one of the wavy lines is α, the other is β]. manufacturing method, formula (wherein R', R ¹ ', R ² ' and X' are respectively the groups R, R ¹ , R ² and X of the above formula, Z is hydrogen and P is hydrogen, with the proviso that R', R ¹ ′,
At least one of R ² ', X', Z and P is a protecting group) is catalytically hydrogenolyzed. 7. The method of claim 6, wherein the protecting group to be removed is selected from the group consisting of benzyl ester, benzyl ether, benzylamino or benzylidene acetal.