JPS61243074A - 2,4-dideoxysialic acid derivative - Google Patents

Abstract

NEW MATERIAL:The 2,4-dideoxysialic acid derivative of formula I [R1 and R2 are independent H or O-acetyl (R1 and R2 are not H at the same time); R3 is carboxyl or methoxycarbonyl; all R4 groups are same and are hydroxymethyl or O-acetyl]. EXAMPLE:Methyl 2-deoxy-2,3-dehydro-4,7,8,9-tetra-O-acetyl-N-acetyl-neuraminate. USE:A medicinal drug capable of activating suppressor T-cell and having immune regulating action to suppress the immunoglobulin production of B-cell. PREPARATION:The compound of formula III is produced by reacting the com pound of formula II with acetic anhydride in the presence of a relatively small amount of concentrated sulfuric acid at about 20-25 deg.C, or the compound of formula IV is obtained by reacting the above components in the presence of a relatively large amount of concentrated sulfuric acid at about 80-90 deg.C.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to immunologically active sialic acid derivatives and methods for their production. Sialic acid is present in sialocomplexes (I! proteins, glycolipids,
It is known to exist as oligosaccharides and polysaccharides.

In recent years, this compound has become important in medical and pharmaceutical fields such as neurological function, cancer, inflammation, immunity, viral infection, differentiation, and hormone receptors, and is attracting attention as a unique active molecule localized on the cell surface.

However, the role played by sialic acid in the sialo complex is still a matter of speculation.

This compound has been further studied by many natural product organic chemists and has already led to various simple derivatives. However, no significant physiologically active derivatives are known yet. The object of the present invention is to obtain a novel compound having excellent physiological activity.

Furthermore, in recent years, the diversification of treatments for malignant tumors of hematopoietic organs, various cancer diseases, collagen diseases, etc. has certainly had a life-extending effect. However, on the other hand, the frequency of use of drugs such as adrenocortical hormones and immunosuppressants cannot be avoided, and as a result, many side effects are occurring as well as a decrease in so-called immunity. The present inventors have focused on sialic acid, which is a component unique to living organisms, and have conducted intensive research into immunomodulators that have fewer side effects and have the ability to regulate the immune surveillance mechanism through chemical modification of sialic acid.
As a result, we have arrived at a novel compound of the present invention that has so-called immunomodulatory effects that activate suppressor T cells and suppress immunoglobulin production by B cells.

That is, the present invention provides (in the formula, R1 and R2 are independently hydrogen or 0-acetyl group, except when both R and R2 are 0-acetyl group, R3 is carboxyl group or methoxy (all R4s are the same and are a hydroxyl group or an O-acetyl group).

The method for producing the sialic acid derivative of formula (1) of the present invention will be explained below.

First, a known compound represented by the following formula (II) was reacted with acetic anhydride and concentrated sulfuric acid under different reaction conditions, respectively, and the formula (I[[
) or a compound of formula (TV') is obtained.

Φ 1-/Σ In the above reaction, a relatively small amount of concentrated sulfuric acid ([■] 3 mm
(approximately 10 to 20 mg per ol), and the reaction temperature was approximately 20
When carried out at ~25°C, a compound of formula (Ill) is obtained, while concentrated sulfuric acid is added in a relatively large amount ([■) about 0 to 3 mmol.
．． 08~0.12g), reaction temperature approximately 80~90℃
It has been found that a compound of formula (IV) can be obtained by carrying out the procedure. Conventionally, V, Kumar et al. used 100 mg of concentrated sulfuric acid for 3 mmol of [■] and carried out the reaction at a temperature of about 50°C, and the resulting reaction products had the formula (III) and the formula (mV
) compound (mixing ratio 1:1), its separation and clear production were difficult, requiring a complicated process of hydrolyzing the product and separating it by high performance liquid chromatography (V
, Kumar et al.

Carbohydr, Res, 94% 123~
l p. 30, 1981). In contrast, according to the method of the present inventors, the compounds of formula (m) and formula (IV) can be easily synthesized separately.

Next, the compounds of formula (III) and formula (IV) above are reduced under various conditions to obtain 2[) of the present invention.

U In the above reduction reaction, the compound of formula (1) is supported on carbon in an alcohol solvent such as methanol to obtain a compound represented by formula (1)-1.

Alternatively, the compound of formula (III) may be mixed with methanol-acetic acid (3:
Using 10% Pd-C as a catalyst in the mixed solution of 1),
A compound represented by formula (1)-2 is obtained by centrifugation at 5° C. in a hydrogen stream for about 2 hours.

Further, when the compound of formula (IV) is reduced in the same manner as above using 10% Pd-C as a catalyst in a methanol-acetic acid mixed solution, a compound represented by formula [1)-3 is obtained.

Next, the compounds of formulas (1)-2 and (1)-3 above were added to about 20-25% of the compound using about 1-2N caustic soda solution, respectively.
C. for about -12 to 20 hours, a compound of the invention is obtained in which the O-7 cetyl group is converted into a hydroxyl group and the methoxycarbonyl group is converted into a carboxyl group.

The compounds of the formulas (III) and (TV), which are intermediates of the compounds of the present invention, are cerium ammonium nitrate-sodium azide or N-promoscinimide-
By reacting with benzyl alcohol, regular azide derivatives and brominated derivatives can be synthesized, the details of which are shown in the reference examples below.

The compounds of general formula (1) thus obtained each have a remarkable immunomodulating effect.

The immunomodulatory effect could be confirmed by the following method.

Effect of ConA on activation of mouse lung lymphocytes: T cells are activated nonspecifically by ConA;
The sialic acid derivative of the present invention was added to this reaction system, and its effect was investigated. Specifically, compounds of ConA and general formula CI) were added to lung lymphocytes (SPC'') obtained from BALB/C mice, and the mixture was incubated at 37°C with 5% C on a microplate.
The cells were cultured for 20 hours while being fed Ot.

Tritium-labeled thymidine was added thereto, and after further culturing at 37°C for 10 hours, spc was collected, and the amount of 3H-thymidine incorporated into SPC was measured using a scintillation counter.

The compound represented by general formula CI) was found to promote and enhance the uptake of 3H-thymidine, and had an enhancing effect on T cell activation by ConA.

Effect on immunoglobulin production by mouse lung lymphocytes: The sialic acid derivative of the present invention, which was shown to have a T cell activating effect in the above experiment, was further investigated for its effect on immunoglobulin production by measuring the number of plaque-forming cells. did. Mazu SPC with sheep red blood cells (SRB C)
and each compound of general formula [1] were added and cultured at 37°C for 5 days. 5RBC and complement were added again to the obtained sensitized SPC.

PFCs (plaque-forming cells) were counted after culturing at 37° C. for 3 to 12 hours in a Cunningham chamber.

A decrease in PFC was observed, and cell viability was equivalent to that of the control standard, confirming an enhanced suppressive effect on immunoglobulin production.

The compounds of the present invention showed activity in the two aforementioned tests. In other words, it is thought that immunoglobulin production was suppressed by activation of suppressor T cells.

Conventionally, in autoimmune diseases such as collagen diseases, a decline in the function of suppressor T cells has been observed.

Therefore, the sialic acid derivative of the present invention having suppressive T cell activating activity is expected to be useful in clinical applications as an immunomodulator.

The present invention will be explained below using examples. These examples are merely illustrative of the invention and therefore, of course, are not intended to limit it.

Example 1 Methyl N-acetylneuraminate 0.97 g (3 m
mol) were added with 20 ml of acetic anhydride and 20 mg of concentrated sulfuric acid, and the mixture was stirred at 20-25°C for 10 hours, then poured into 100 ml of ice water and neutralized with NaHCO2.

The neutralized reaction solution was extracted with chloroform, washed with water, dried with Glauber's salt, filtered, and then the solvent was distilled off to dryness.

The obtained crude reaction product was dissolved in 30 ml of dry methanol, treated with 0.5 g of sodium methylate, allowed to stand at room temperature for 1 hour, neutralized using Dowex-50 (H"), and filtered through the mouth. , wash the resin thoroughly with methanol.

When the mouth plate and cleaning solution are concentrated, crystals will precipitate. The crystals were recrystallized from methanol, treated with 5 mff of acetic anhydride, and 4 ml of pyridine, kept at room temperature overnight, poured into ice water, extracted with chloroform, 1N-HCl 5% Na HCO3, Ym Japanese saline, and dried with Glauber's salt, the solvent was distilled off, and the mixture was dried. The title compound was then obtained as a colorless powder. Yield 0.65 g (46% yield) (α) +40° (C=1, in MeOH)
Mass m/z 474 (M” +1) Elemental analysis Calculated value with C2゜Hz, NO+□ and 17 C:
50.74, H: 5.75, N: 2.96 Actual value C: 50.40, 'H: 5.65, N
: 2.82'HNMR (400MHz, CDC#3
Medium) δo (TMS) 1.920s, 3) 1.
NAc).

2.052.2.064, 2.075, 2.126 (1
2tl, 4xOAc).

3.800 (s, 3H, COOMe).

4.197 (dd, 11. J=7.1. -1
2.5Hz, 9-11) 4.382 (broad
dd, 18. J=9.9. 2.0Hz,
5-H) 4.380 (broad s, 18.6
-H)4.654(dd, LH,J=-12,5,
2,93Hz, 9-H')5.321(ddd,
LH, J=7.1.2.9.2.0Hz, 8-H)5
．． 402 (broad dd, 18. J=2.
0Hz, 7-)1) 5.48Hdd, IH,J=
2.9.2.0Hz, 4-H)5.987(d,
LH, J=2.9Hz, 3-H)6.086(d
, IH, J=9.9Hz, NH)rRν□x
(cm-9 1740 (OAc, COOMe), 165B (
amidel) 1535 (amideIt) Example 2 Methyl N-acetylneuraminate 1. Og(3,09
20 mA' of acetic anhydride and 0.1 g of concentrated sulfuric acid (mmol) were added, stirred at room temperature until dissolved (about 30 minutes), and then
The mixture was stirred at -90°C for 3 hours. The reaction solution was poured into ice water, neutralized with NaHCO3, extracted with chloroform, and the extract was dried over Glauber's salt for 13 passes, and the solvent was distilled off to dryness. The remaining reaction product was crystallized using isopropyl alcohol and recrystallized to obtain colorless needle-like crystals of the title compound. Yield 1.24g (yield 85%) Melting point 1
00-103℃ Mass m/z 474 (M” +1) (α)
D 14'5° (C=1, in MeOH) Elemental analysis C2°H2? Calculated value as N0I2 C; 50.
74, H, 5,75, N, 2.96 actual value C;
50.55, H; 6.04, N; 2.71'H
-NMR (300 MHz, in CDCjt x) δH (T
MS) 1.958 (s, 3H, NAc) 2.0?9.2.103, 2.120. 2.127(
121,4xOAc)3.816(s, 3H,COO
Me).

4.205 (dd, 18. J=12.5.7.5H
z, 9-H) 4.298 (dd, 1B, J=11
．． 0.2.1Hz, 6-H) 4.322(dt, L
H, J=4.3, 11.0Hz, 5-II) 4.80
3 (dd, IH, J= 12.5, 2.7Hz,
9-H') 5.162 (dd, IH, J=5.
6.4.3ffz, 4-H)5.322(ddd,
1)1. J=4.1, 7.5, 2.7Hz,
8-H) 5.511 (dd, IH, J=4.1,
2.1) r, z, 7-H) 5.615 (d, L
H, J=10.0Hz, NH)-6,233(d,
IH, J=5.6Hz, 3-H) 1750 (-0A
c), 1738 (-0Ac COOMe) 1658
(amide I), 1535 (amide II) Example 3 98 mg of the compound obtained in Example 1 (0,2 Immo
l) with 20mj of methanol! After absorbing 5 ml of hydrogen at normal pressure and room temperature using PtOzlOOmg and PtOzlOOmg, the reaction solution was passed through the mouth and the solvent was distilled off to dryness. Next, the reaction product was separated and purified by silica gel column chromatography using a developing solvent of chloroform:methanol=ioo:t to obtain a colorless powder of the title compound. Yield 17 mg (yield 20%) (α)D +2° (C=1, in MeOH)Mass
m/z 416 (M” +1) Elemental analysis Cr
Calculated value as s Hz s N OIo C; 52.
05, H; 6.02, N; 3.37 actual value C;
52.06, H, 6,15, N; 3.29'H-N
MR (400MHz, in CDCl x) 6M (TMS)
1.958 (s, 3H, NAc) 2.090, 2,066.2.115 (9H, 3xOA
c) 2.197 (ddd, J=20.3, 4.5.4
．． 1fLz, 4-H) 2.62Hddd, J=20
．． 3.5.6B, 4. IH2,4-11') 3.785
(s, IL COOMe).

4.177 (t, IH, J=5.9)1. z, 6-
H) 4.247 (d, IH, J=7.3, 15.6H
z, 9-H)4.365(dddd, J=8?6,
4.5.5.9,4. 'llz, 5-H) 4.56H
dd, IH, J= 3.5, 15.6Hz, 9-H
')5.233(ddd, 1)1. J=3.9,7
．． 3,3.5Hz, 8-tl)5.440(dd,
IH, J=5.9, 3.9Hz, 7-H) 5.645
(d, IH, J=8.6Hz, N)l) IR sil, 1. XcI11-1 1730 (C0OHJ+ 1640 (amide I)
.

1525 (amide II) Example 4 100 mg of the compound obtained in Example 1 (0.21 m
10% Pd-C100m in 20ml MeOH
A catalytic reduction reaction was carried out using g for 2 hours at normal pressure and room temperature,
The reaction solution was filtered through the mouth, the solvent was distilled off, and the mixture was dried. The reaction product was crystallized from ether and recrystallized to obtain colorless needle-like crystals of the title compound. Yield: 75 mg (yield: 75%) Melting point: 185-186°C (α) OH6° (C=1, in M e OH) Mass
m/z 476 (M” +1) Elemental analysis C
Calculated value as 2゜H2, NO, □ C; 50.53,
H: 6.11, N: 2.95 Actual value C: 50.49
, H; 6.14, N; 2.8B' HNMR (40
0MHz, CDC1:l) δ14 (TMS)1.
858 (dt, LH, J=12.1, 11.2112
．． 3-H)1.892(s, 38.NAc)2
．． 043, 2.072, 2.141 (12H, 4xOA
c) 2.406 (ddd, IH, J=12.1.4.
9.2.4112.3-H')3.699(dd, I
H, J=2.2, 10.411z, 6-H) 3.77
1(s, 3L-COOMe).

4.021 (q, LH, J=10.4Hz, 5-H
) 4.056 (dd, 18. J=12.2, 2.4
Hz, 2-H) 4.143 (dd, IH, J・7.
3,12.5H,z, 9-fl)4.650(dd,
In, J=2.6, 12.5Hz, 9-H')5
．． 046(ddd, 1)1. J=11.2,5.0
,10.0Ilz, 4-)1)5.250(ddd,
18. J, 2.4.7.3, 2.6Hz, 8-H
) 5.362 (dd, LH, J: 2.2, 2.4Hz
, 7-8) 5.385 (d, LH, J=9.2H2
, NH) 1730 (-0Ac, -COOMe).

1650(amide l)+1530(amide
II) Example 5 0.20 mg (0.42 m
mol) to MeOHl 5rnj! , AcOl(5
mj! A catalytic reduction reaction was carried out for 2 hours at normal pressure and room temperature using 10 g of 10% Pd-Co in
After evaporation of the solvent and drying, the remaining reaction product was crystallized using ether and recrystallized to obtain colorless needle-like crystals of the title compound. Yield 149 mg (yield 85%) Melting point 1
61-162℃ [α]. +15° (C=1. in MeOH) Mass
m/z 417 (M”) Elemental analysis Cl1lH
2, Calculated value as No. 10 c; 51.79, H;
6.52, Ni3.36 actual value C, 51,47, H
, 6,39,N; 3.44'H-NMR (300MHz
, CDC12, middle) δll (TMS) 1.517 (dd
d, LH, J=12.6, 3.42.12.5Hz,
4-Hax) 1.723 (ddd, LH, J=10.3
, 12.5.12.6Hz, 3-Wax) 1.894 (
s, 3H, NAc) 2.021, 2.049, 2.111 (9H, 3xOA
c) 2.0-2.2 (3-Heq, 4-Heq) 3
．． 556 (dd, ill, J=10.3,2.
4Hz, 6-H) 3.732(s, 311.
-COOMe).

3.912 (dddd, IH, J=12.5.3.
5, 10.7, 10.3Hz.

5-■) 3.913 (dd, IH, J=2.0, 10.3H
z, 2-H) 4.141 (dd, IH, J=7
．． 3.12.2H2,9-tl)4.685(dd,
IN, J=2.4.12.2Hz, 9-1f
') 5.234 (ddd, LH, J=4.4, 7.
3,2.4Hz, 8-H)5.369(dd,
LH, J=2.4.4.4Hz, 7-105.31
0(d, LH, J=10.78Z, NH)17
40 (-08c, -COOMe).

1658 (amide I), 1545 (amide
(2) Furthermore, a catalytic reduction product of the compound obtained in Example 2 was obtained using PLO□ in methanol-acetic acid in the same manner as above.

Example 6 100 mg of the compound obtained in Example 5 was added to 1 ml of MeOH.
Add 1 ml of IN-Na OH, leave at room temperature for 15 hours, add 5 ml of water, and add Dowex-50 (H
The reaction solution was neutralized using a solution of 100% sodium chloride, filtered, and the solvent was distilled off to dryness, followed by crystallization and recrystallization using EtOH to obtain colorless prismatic crystals of the title compound. Yield: 43n+g. (
yield 65%), melting point 209-210°C (tx]I) -29° (C = 0.5, M e OH
Middle) Elemental analysis CIIHI9NO? Calculated values as C: 47.65, H: 6.86, N
: 5. (15 Actual value C: 47.11, H: 6
．． 53, N: 5.24'tlNMR (60MHz
, DzO) δH(DSS)2.10(s, 38
．． NAc)IRshi11. XcITl-1 1725(-COOH).

1630(amide I)+1540(amide
n) Reference Example 1 Compound 500B obtained in Example 2 (1,06 mmo
Add dry acetonitrile Qmf to l) and add NaN3
(120 mg) was added thereto, and the mixture was stirred at 0°C under a nitrogen stream.

Cerium ammonium sulfate (NH4) ice (NOz)i (1.8 g) was added to this, and stirring was continued. After 10 hours, the reaction solution was poured into ice water and extracted with ether. The ether layer was dried with Glauber's salt and then concentrated under reduced pressure. Colorless needle-like crystals of the title compound were obtained. (Yield: 88.2%), melting point 135-136°C IR (KBr) ν, , x am-' 2
125(-Nz)1650(-ONO□) Reference Example 2 100 mg of the compound obtained in Reference Example 1 (0,1711
aol) in dry methanol, NaOMe5mI
After stirring at room temperature for 30 minutes, acetic acid was added to neutralize and the solvent was distilled off. Thereafter, 5 ml of acetic anhydride and 5 ml of pyridine were added, and the mixture was allowed to stand overnight for acetylation.

After pouring ice water into the reaction solution to decompose excess acetic anhydride, extraction with chloroform was performed. After drying the chloroform layer with Glauber's salt and distilling off the solvent, a pyridine odor was emitted, so ethanol was added and azeotroped. Colorless needle-like crystals of the title compound were obtained. (Yield 50%) Melting point (decomposition) 211°C (ff)o+4.8° (C=0.33, in MeOH)
IR(KBr) ν11. ．． Cf1-' 2125
(-Nff) Elemental analysis C2OH2? Calculated value as N5OI6 C: 41.59, H: ・4.68, N
: 12.13 Actual value C: 40.58, H: 4
．． 46, N: 11.56 Reference Example 3 Sufficient amount of 2-deoxy-2,3-dehydro-4,7,8゜9-tetra-0-acetyl-N-acetylneuraminic acid methyl ester (473 mg, 1 mmol) After drying, dissolve in 29 ml of methanol that has been distilled and treated with 3 molecular sieves, add NBS (405 mg),
The mixture was stirred at room temperature for 3 hours. After distilling off the solvent of the reaction solution, it was dissolved in a small amount of chloroform, and excess NBS was precipitated as crystals, which were then gravity filtered. A yellow oil was obtained. Purification was performed using a Rover column. C solvent: chloroform) The title compound was separated as colorless needle-like crystals.

(117 mg, yield 20%) Melting point 144-147°C [α]D -65,5° (C: 0.94, MeOH)
IRv,,, am-' 1750 (-COO
Me), 1660 (-NHCO-) 1550 (-NHC
O-) Mass (M") mHz 524 (M"-
OAc)526(M''-08C) NMR(300MHz, CDCj?3) δo
(TMS) 3.849ppn+(3H,s, C
OOMe) 3.537 (3H,s, OMe)
2.144 (3H,s, 0Ac)2.08
7 (3B, s, 0Ac)2.075
(311,s, 0Ac)2.037 (38
,s, 0Ac)1.909 (3H,s,
N! ! 3.983 to 8C (IH+cLJz, a
・10.75. H-3) Reference Example 4 After sufficiently drying 2-deoxy-2,3-dehydro-4,7,8゜9-tetra-0-acetyl-N-acetylneuraminic acid methyl ester (473 mg, 1 mmol) , distilled and treated with 3 molecular sieves 575 mg of benzyl alcohol dissolved in 20 ml of acetonitrile and treated with 3 molecular sieves 5 NBS (2
85 mg) was added thereto, and the mixture was stirred at room temperature for 4 hours.

After distilling off the solvent of the reaction solution, the NBS precipitate was removed by gravity filtration. Silica gel column (developing solvent: benzene-
Purification was carried out with acetone (l O/1). Repurification was performed using a Rover column.

(Silica gel, developing solvent chloroform-methanol (8
0/1)) The title compound-1 was isolated as colorless needle-like crystals.

(20 mg, yield: 3.03%) Melting point 175-178°C [α]. ±5.8° (C=0.35, MeOH)
rRc-x-' 17ff5(-COOM
e), 1655(-NHCO-)0m 1550(-NHCO-) NMR(400MH2, CDC13) δH(TM
S) 3.73109 (3) i, s, COOMe) 2゜1
8926 (3H, s, 0Ac) 2.07838 (3
H, s, 0Ac) 2.04337 (3B, s,
0Ac) 1.95054(3f(,s, 0Ac
) 1.91159 (3H, s, NHAc) 4.64
863 (IL d, J=11.54.-CH-〇)
4.41495(1)1. d, J=11.54. -C)
I-Q) Showa year, month, day 1, case description 1985 Patent Application No. 69742 2, title of the invention 2.4-dideoxysialic acid derivative 3, person making the amendment Relationship with the case Applicant name Title Kanto Ishikawa Pharmaceutical Co., Ltd. Co., Ltd. 4, Agent 6, Subject of amendment: Detailed explanation of the invention in the specification column 7, contents of the amendment Add the following sentence following the last line of page 27 of the specification cylinder 0 "Reference Example 3 Codexico , 3-dehydro-'A7.g,9-tetra-0-7cetyl-N-acetylneuraminic acid methyl ester (4731t7,/mmoJ) was dried using P2O5, dissolved in methanol 20- 4t
) was added, and the mixture was stirred at room temperature for 3 hours. After distilling off the solvent of the reaction solution, dissolve in a small amount of chloroform and remove excess N.
The BS was crystallized and filtered. A yellow oil was obtained. Purification was performed using a Roper 7 ram (Merck Grees).

, solvent: chloroform) The above α-glucosyl body, β-glucosyl body, and raw material were isolated. α-glucosyl form and β
- The glucosyl body honeychill was crystallized.

α-Glucosyl substance melting point: /14t-/17°C (decomposition) Colorless needle crystals (yield: /6%) [α11;7:-4! , t'(c: θ94. M4!0
H) IR':4F crIL-'/7,! i-
0 (-COOMe) / 6 Otsu 0 (-NHCO-) / Tata 0 (-NHCO-) Mass (M+) rry'z! r, 2lI
t(M"-0Ac)52(M-OAC) NMR(3θQMHz, cocg3)aH(TMS
)3, gl19ppm (3H, S, COOMe)
3JJ 7 (jH, s, OMe) 21'14
t(3H,s,0Ac)2gg7(JH,s
, 0Ac) 5 (jh, s, oAc) 2.037 (, aH, s-, 0Ac) i90q
(jH,s,NHAc), 2.9g3 (j
H, d, J3. Gu=/θ7! , H-3)! f, 4' q
2 (jH, t, J3,4 =IOJO-%5
=10JO.

H-da) gu, 2 Otsuri (/H, dt-Jq3 ='
/θj O, J4,4 = 10JO−j Hu□=/
θ5θ, H-5) tAA! rg (jH, dd, J↓6 =/θA
;0-J6ri=2.20゜H-6)ri, 2Ari (jH, dd, J47=Yuko0,
J7. g = g, 79.

H-7) Fu3! ;3 (jH, ddd, Jpe9=對-”g
, 9'=39H-4) 'AO419(/)'-"'L-'?9#=/2.11
4tH-9) lA collO (#+, dd, H-y') elemental analysis Calculated value as C21H5ONO13Br C; 4
',? ,/Otsu, H;yo/7. N; 2.3W Actual Measured Value C; 1I, 3.3* H; Left/Right, N;
Melting point of 2-4t β-glufucil body, 170-172°C Colorless needle crystals (yield/q%) [α 4: +/&4° (C: / , MeOH) 71
1m −1 1Rνmax cMFl / 73QC/ -COO
Me)/is 70 (-NHCO-)/! r, 3. ! ; (-NHCO-) Mass(M)r$z 3;24t (M-0A
C)! r,, 2is (M -OAc) NMR (lIooMHz, cock3) aH (vM
s)3gll! rpm (3H,s, cooMe
)3,. 300 (3H, s, OMe) L/g
'l (3M, s, 0Ac) YuOg, 3 (3
H,s,OAc)QOg/(,aH,s,
0Ac) θJ g (J He s -0Ac)
19/g (3H,s,NHAc)'AA16
(/HedeJ3.Da=3゜Otsu7.H-J')! ;,'
170 (jH, dd, %1/QA7.H-4')
'A3! r! r (/H-broad Q-
'j,NH9,4'To -j part seasonal θ! r, 3. H-!
r) lAogko (jH, dd, J67=ig'l, H-A
)Left J! rg'(jH, dd,, j=da,!-g,
H-7) 2g left 99 (jH, ddd, Jro q=7.'1g-
JBon9. =2. Dada.

H-za44/g7 (jH, dd, Jqqt=/2.3/
, H-9) Showa year month day 1, case description 1985 patent application No. 69742 2
, Title of the invention 2.4-dideoxysialic acid derivative 3
, Relationship with the case of the person making the amendment Applicant name Kanto Ishi Pharmaceutical Co., Ltd. 4, Agent 6, Subject of amendment Detailed explanation of the invention in the specification column (1) Procedural Amendment No. dated June 26, 1985 Add the following sentence following "...H-9')" on the last line of page 5.

[Reference Example 6 Compound obtained in Reference Example 1 (50 mg, 0.087 mm
ol), sodium acetate (1 g) and acetic acid (20
mg) was added thereto, and the mixture was heated under reflux at 100°C for 1 hour. Water was added to this reaction solution and extracted with chloroform. The chloroform layer was washed with water, dried over sodium sulfate, and then the solvent was distilled off. A yellow oil was obtained. Roper column (silica gel, M
erck, Gr5Be B).

(Developing solvent: chloroform/methanol (''/,)) White crystals were obtained. (Yield 20%) ((Z) + 48.9° (C = 0.18. MeOH)
L Rv Cm-' 1740 (-C[JUM
e) 1640 (-NHCO-) 1540 (-NHCO-) 2100 (-N3)

Claims (1)

[Claims] General formula ▲ Numerical formula, chemical formula, table, etc. ▼ [ I ] (In the formula, R_1 and R_2 are independently hydrogen or an O-acetyl group, provided that both R_1 and R_2 are both O- (except when it is an acetyl group, R_3 is a carboxyl group or a methoxycarbonyl group, and R_4 are all the same and are a hydroxyl group or an O-acetyl group).