JPH08217785A - Sulfated fluoroalkyl laminaripentaoside and antiviral agent containing the same - Google Patents

Abstract

PURPOSE: To obtain the subject new compound, having high fat solubility and useful as an antiviral agent, etc., effective against an HIV by fluoroalkylating hydroxyl group at a specific site in a laminaripentaose and sulfating a part or all of the residual hydroxyl groups. CONSTITUTION: This new fluoroalkyllaminaripentaoside sulfate (salt) has hydrogen atom of hydroxyl group at the 1-position in a reducing terminal saccharide of a laminaripentaose substituted by a fluoroalkyl group such as the formula C2 H4 C8 F17 , C2 H4 C10 F21 , C6 H12 C4 F9 or C6 H12 C8 F17 , a part or all of the residual hydroxyl groups substituted by sulfuric ester groups and high fat solubility and is useful especially as an antiviral agent, etc., having excellent actions against a human immunodeficiency virus (HIV). The compound is obtained by reacting a D-laminaripentaose peracetate with a compound of the formula C10 F21 C2 H4 OH in the presence of tin tetrachloride in a solvent, then removing the acetyl group and further reacting the resultant compound with a sulfur trioxide pyridine complex.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel oligosaccharide derivative having a fluoroalkyl group at the 1-position of a reducing terminal sugar and a novel antiviral agent obtained by sulfating this, and particularly to HIV (Human). Immunodef
The present invention provides a preventive or therapeutic drug for infectious diseases caused by iciency virus).

[0002]

BACKGROUND OF THE INVENTION As a derivative of an oligosaccharide by the present applicant, a compound having an aliphatic alkyl group (JP-A-5-78382) and a compound derivatized with an aromatic compound having an alkyl group have been known. (JP-A-5-27938)
1). On the other hand, there was no example of an oligosaccharide derivative having a fluoroalkyl group which is considered to have interesting properties as an alkyl group.

AZT has been used as a therapeutic agent for AIDS.
Nucleic acid analogs such as (azidothymidine) and ddI (dideoxyinosine) are approved drugs, but side effects and
The emergence of resistant patients has become a problem. Recently, the anti-HIV action of sulfated saccharides and their derivatives has attracted attention. For example, as an anti-HIV effect of sulfated polysaccharides, JP-A-62-215529; Hideki Nak
asima et al Jpn. J. Cancer
Res. (Gann) 78,1164-1168, (1
987); Osamu Yoshida et al.
Biochemical Pha-rmacolog
y Vol. 37, 2887-2891 (1988);
There is JP-A-1-103601 and JP-A-3-43401 for further substituted polysaccharides. Further, as an oligosaccharide derivative (Japanese Patent Laid-Open No. 5-78382) is known.

[0004]

DISCLOSURE OF THE INVENTION The present invention relates to an oligosaccharide derivative having a fluoroalkyl group, which has not been reported so far, and includes a fluoroalkyl oligosaccharide glycoside peracetate and a sulfated fluoroalkyl oligosaccharide conjugate. To provide glycosides.

Furthermore, the present invention further improves the hydrophobicity of the sulfated alkyl oligosaccharides by introducing fluorine into the alkyl group, as compared with the sulfated alkyl oligosaccharides which are expected to act by a mechanism different from that of the conventional sulfated polysaccharides. The present invention relates to providing an antiviral agent which enhances the bioavailability of a sulfated alkyl oligosaccharide derivative.

[0006]

Means for Solving the Problems The present inventors have found that a sulfated product of an oligosaccharide alkyl ether, which is a compound having an extremely low molecular weight, exhibits anti-HIV activity as compared with the above-mentioned sulfated polysaccharide. Focusing attention, as a result of earnest research, a novel fluoroalkyl oligosaccharide in which fluorine is further introduced into the alkyl group portion of the oligosaccharide alkyl ether sulfate to improve lipophilicity has antiviral activity and is a synthetic precursor thereof. We have found that the oligosaccharide fluoroalkyl glycoside peracetate is a useful intermediate.

That is, in the sugar moiety, five glucoses are β-
1,3 glycoside-bonded laminaripentaose, in which the hydrogen atom of the hydroxyl group at the 1-position of the reducing terminal sugar is replaced by a fluoroalkyl group, and the remaining hydroxyl groups are partially or entirely replaced by a sulfate ester group. And a sulfated fluoroalkyl laminaripentaoside or a physiologically acceptable salt thereof and an antiviral agent containing the same as an active ingredient. Further, it relates to a peracetylfluoroalkyl oligoglycoside in which the hydrogen atom of the hydroxyl group at the 1-position of the reducing terminal sugar of laminaripentaose is substituted with a fluoroalkyl group, and all the remaining hydroxyl groups are substituted with acetyl groups.

First, a sulfated fluoroalkyl laminaripentaoside will be described as the first compound of the present invention. The oligosaccharide moiety of this compound is laminaripentaose. In the compound of the present application, the hydrogen atom at the 1-position of the reducing terminal sugar of laminaripentaose is substituted with a fluoroalkyl group. The fluoroalkyl group which is an aglycone may have a straight chain or branched chain having 22 or less carbon atoms in the alkyl chain, and may be a perfluoroalkyl group in which all the hydrogens of the alkyl group are fluorinated or partially. Fluorinated fluoroalkyl groups may be used.

Regarding the fluoroalkyl group moiety of the compound of the present invention, the bonding position thereof is important, and the fluoroalkyl group must be bonded to the 1-position of the terminal reducing sugar in the oligosaccharide moiety. However, the configuration may be such that the fluoroalkyl group is α-bonded or β-bonded to the sugar. Fluoroalkyl group has 1 carbon
0-14 are preferable,

[0010]

Embedded image _{-C 2 H 4 C 8 F 17} , -C 2 H 4 C 10 F 21, -C 6 H
A fluoroalkyl group selected from the group consisting of ₁₂ C ₄ F ₉ or —C ₆ H ₁₂ C ₈ F ₁₇ is more preferable, and especially

[0011]

Embedded image Fluoroalkyl groups represented by —C ₂ H ₄ C ₁₀ F ₂₁ are preferred.

The sulfated fluoroalkyl laminaripentaoside of the present invention is a reducing terminal sugar 1 of laminaripentaose.
A compound in which some or all of the hydroxyl groups other than the positions are substituted with a sulfate group. The higher the sulfation rate, the better
It is desirable that 70 to 100%, more preferably 90 to 100%, of the substitutable hydroxyl group is substituted with a sulfate ester group.

The present invention also includes a compound in which the sulfate ester group is a physiologically acceptable salt. Examples of the physiologically acceptable salt include sodium salt, potassium salt, magnesium salt and the like.

The second compound of the present invention, peracetylfluoroalkyllaminaripentaoside, will be described below. The oligosaccharide moiety of this compound is laminaripentaose, and the hydrogen atom at the 1-position of the reducing terminal sugar of this laminaripentaose is substituted with a fluoroalkyl group. The fluoroalkyl group that is an aglycone is the same as that described for the sulfated fluoroalkyl laminaripentaoside, and preferably has 10 to 14 carbon atoms,

[0015]

Embedded image _{-C 2 H 4 C 8 F 17} , -C 2 H 4 C 10 F 21, -C 6 H
A fluoroalkyl group selected from the group consisting of ₁₂ C ₄ F ₉ or —C ₆ H ₁₂ C ₈ F ₁₇ is more preferable, and especially

[0016]

Embedded image Fluoroalkyl groups represented by —C ₂ H ₄ C ₁₀ F ₂₁ are preferred.

The peracetylfluoroalkyl laminaripentaoside of the present invention is a compound in which all of the hydroxyl groups other than the 1-position of the reducing terminal sugar of laminaripentaose are substituted with acetyl groups.

More specific examples of the compounds of the present invention are shown below. Although the hydroxyl groups of sugars have been described as non-derivatized compounds, the compounds of the present invention have the skeleton substituted with the above-mentioned sulfate ester group or acetyl group. 1H, 1H, 2H, 2H-perfluorodecyl β-D-glucopyranosyl- (1 → 3)-{β
-D-glucopyranosyl- (1 → 3)} ₃ -β-D-glucopyranoside, 1H, 1H, 2H, 2H-perfluorododecyl β-D-glucopyranosyl- (1 → 3)-
{Β-D-glucopyranosyl- (1 → 3)} ₃ -β-D
-Glucopyranoside, 1H, 1H, 2H, 2H, 3H,
3H, 4H, 4H, 5H, 5H, 6H, 6H-perfluorodecyl β-D-glucopyranosyl- (1 → 3)-
{Β-D-glucopyranosyl- (1 → 3)} ₃ -β-D
-Glucopyranoside, 1H, 1H, 2H, 2H, 3H,
3H, 4H, 4H, 5H, 5H, 6H, 6H-perfluorotetradecyl β-D-glucopyranosyl- (1 →
3)-{β-D-glucopyranosyl- (1 → 3)} ₃ −
β-D-glucopyranoside.

Next, a method for producing the sulfated fluoroalkyl laminaripentaoside and peracetyl fluoroalkyl laminaripentaoside of the present invention will be described. As a method, in the method for producing an alkyl oligosaccharide described in JP-A-5-78382, the saccharide can be produced by using laminaripentaose and fluoroalcohol in place of alcohol. That is, a method in which sugar peracetate is reacted with fluoroalcohol under acidic conditions, sugar peracetate
enigs.Knorr) method to form glycosyl bromide and then reacting with fluoroalcohol using silver salt or silver salt, method of reacting glycosyl bromide with fluoroalcohol in the presence of tetraethylammonium bromide, Any method such as a method of reacting sugar imidate with fluoroalcohol can be used.
Of these, the method of reacting fluoroalcohols under acidic conditions is the simplest.

Further, the perfluoroacetate of the fluoroalkyl oligosaccharide obtained as described above is deacetylated with sodium methoxide, ammonia or the like by a conventional method to convert it into a fluoroalkyl oligosaccharide in which a hydroxyl group is not substituted. it can. Based on this fluoroalkyl oligosaccharide in which the hydroxyl group is not substituted, it can be converted into various derivatives by substituting the hydroxyl group by a conventional method.

Further, a sulfated fluoroalkyl oligosaccharide can be synthesized by sulfating the hydroxyl group of the sugar with a sulfur trioxide pyridine complex, 1-piperidinesulfonic acid, chlorosulfonic acid or the like as a sulfating agent.

Next, the manufacturing method of the present invention will be described in more detail. Although it is manufactured by the following four steps, it is not limited to this method. Furthermore, the fluoroalcohol used as the raw material is Daikin Chemical Products Co., Ltd. or PCR In
Compounds sold under the corporate standard can be used.

(Step 1) Acetylation The hydroxyl groups of oligosaccharides are acetylated by a conventional method. That is, oligosaccharides can be acetylated with sodium or potassium acetate and acetic anhydride.

(Step 2) Fluoroalkylation The acetylated oligosaccharide produced in Step 1 and the fluoroalcohol can be produced by the reaction of an alcohol described in JP-A-5-78382. For example,
The oligosaccharide peracetate prepared in step 1 is reacted with fluoroalcohol in the presence of tin chloride (〓) in a solvent such as methylene chloride or 1,2-dichloroethane or hexafluoro-meta-xylene. At this time, the molar ratio of oligosaccharide peracetate, Lewis acid catalyst and fluoroalcohol is 0.1 to 2.0 molar equivalents of Lewis acid catalyst to oligosaccharide peracetate, preferably 0.5.
˜2.0 molar equivalents, and further 1.0 to 5.0 molar equivalents of fluoroalcohol, preferably 1.0 to 3.0 molar equivalents. The reaction of a mixture of these oligosaccharide peracetates, a Lewis acid catalyst and a fluoroalcohol is carried out in an inert gas from room temperature to the boiling point of the solvent, more preferably 20.
It is carried out at -80 ° C, preferably in the presence of molecular sieves. Although the reaction time varies depending on the oligosaccharide and fluoroalcohol used in the reaction, it is generally 2 to 30.
The time is usually 3 to 15 hours. After the reaction, the crude product obtained through extraction, washing and dehydration operations is purified by silica gel chromatography, and if necessary, reprecipitated and recrystallized to produce a purified product. With such a production method using a Lewis acid, a β-type anomer can be mainly obtained.
Further, the α, β anomer ratio can be adjusted by selecting the production method and the catalyst or operating the purification step.

(Step 3) Deacetylation The fluoroalkyl oligosaccharide peracetate produced in Step 2 is dissolved in an alcohol such as methanol, and a solution of sodium methoxide in methanol is added thereto, and generally 1 to 50 hours, usually 1 Stir at room temperature for ~ 24 hours to perform the deacetylation reaction. The sodium methoxide used in the deacetylation is preferably 0.01 to 1 equivalent to the acetyl group of the sugar, and more preferably 0.
Use 1 to 1 equivalent. After the reaction, sodium ions are removed using a hydrogen ion type cation exchange resin, and then the desired fluoroalkyl oligosaccharide is obtained. Furthermore, if necessary, it can be purified by reprecipitation, recrystallization or the like. Also,
Here, it is also possible to use ammonia or sodium hydroxide instead of sodium methoxide.

(Step 4) Sulfation Although various methods are known for sulfation, any method may be used. Here, a method using a sulfur trioxide pyridine complex will be described.

The fluoroalkyl oligosaccharide is dissolved in pyridine, and a necessary amount of sulfur trioxide pyridine complex is added to the sugar hydroxyl group. 1.2 to complete sulfation
It is preferred to use 3.0 equivalents of sulfur trioxide pyridine complex. The reaction is carried out in an inert gas at room temperature to 100 ° C, preferably 50 to 85 ° C for 0.5 to 72 hours, preferably 0.5 to 2 hours, and dissolved in an appropriate amount of ion-exchanged water at room temperature. Then, a barium hydroxide aqueous solution is further added to adjust the hydrogen ion concentration to 7 to 8. The precipitate formed at this time is removed by centrifugation. The solution after precipitation removal is passed through a sodium ion type cation exchange column column, the column eluate is concentrated and dried to dissolve in ion exchanged water, and an organic solvent such as acetone is added to reprecipitate the resulting precipitate. Is again dissolved in ion-exchanged water and freeze-dried to obtain the desired sulfated fluoroalkyl oligosaccharide.

Further, instead of neutralizing with barium hydroxide, the hydrogen ion concentration is adjusted to 9 to 10 with an aqueous sodium hydroxide solution, concentrated to dryness under reduced pressure at 40 ° C. or below, and subjected to an ultrafiltration method, a reverse osmosis membrane method, It is also possible to separate the excess inorganic salt with a microacylizer (electrical ion exchange membrane manufactured by Asahi Kasei Co., Ltd.) or the like.

In the case of 1-piperidine sulfonic acid, the desired product can be obtained by almost the same treatment. Further, in the present invention, among the above fluoroalkyl laminaripentaosides, a compound having a sulfated oligosaccharide moiety as a water-soluble group and a fluoroalkyl chain as a fat-soluble group is synthesized, and it has extremely good anti-HIV activity. It was a finding.

Although the detailed mechanism of action of the sulfated fluoroalkyl laminaripentaoside of the present invention as an antiviral agent is not clear, it inhibits the adsorption process of viruses to target cells as well as sulfated alkyl oligosaccharides. It is clear that they are doing it. The antiviral agent of the present invention can be orally and parenterally administered in a usual formulation, for example, tablets, capsules, granules, pills, solutions, syrups and the like. As a means for this purpose, the conventional excipients and additives used for producing pharmaceutical products can be used. Examples of conventional excipients include water, physiological saline, alcohol, polyethylene glycol, glycerol ester,
Examples include gelatin, carbohydrate magnesium stearate, talc and the like. Also, as a conventional additive, a preservative,
Examples include sterilizing agents, lubricants, coating agents, wetting agents, emulsifying agents, coloring agents, masking flavors, and aromatic agents.

The dosage of the antiviral agent according to the present invention varies depending on the dosage form of the agent, the number of administrations, the condition of the patient, the weight of the patient, and the severity of the disease.
0.1 mg to 200 mg, preferably 0.5 m
It is preferable to administer g to 100 mg in several divided doses a day. The frequency of administration is also determined by the form of the drug, the condition of the patient, and the weight of the disease. In some cases, continuous intravenous infusion is also possible.

Further, the drug of the present invention has higher lipophilicity by introducing a fluoroalkyl group than in the case of the conventional sulfated alkyl oligosaccharides. Generally, it is said that the higher the lipid solubility of a drug, the higher its bioavailability and the better the uptake of the drug into the body upon oral administration. The fluoroalkyl group in the present invention is more lipophilic than the conventional oligosaccharide derivative due to the effect of the introduced fluorine atom, and as a result, has high bioavailability. For this reason, it is a drug that has higher absorbability upon oral administration than conventional oligosaccharide derivatives and is more easily administered orally as an antiviral agent.

The compounds of the present invention show activity against a wide range of viruses, but the activity against HIV is illustrated as an example. The following test methods are based on R. Pauwe
ls) et al., J. of Biological Met
hods Vol. This method is accepted as an anti-HIV test as shown in 20 309-321 (1988). The activity test was carried out in a 96-well microtiter plate with various concentrations of test substances together with HIV-infected MT.
-4 cells (2.5 x 104 / well, MOI: 0.0
1) is added immediately after infection. In order to know the cytotoxicity of the test substance against MT-4, virus-uninfected cells are likewise incubated with various concentrations of the test substance. After culturing at 37 ° C. for 5 days in a CO ₂ incubator, the number of viable cells is measured by the MTT method. The antiviral activity is a concentration (EC ₅₀ ; 50% ef) at which 50% of cytotoxicity caused by HIV infection is protected.
50% cytotoxicity (CC ₅₀ ; 5) depending on the test substance.
0% cytotoxic concentratio
n) respectively evaluated. These values include EC ₅₀
The smaller the value is, the higher the activity is, and the larger the CC ₅₀ is, the safer the drug is with less toxicity.
This time, the sulfated fluoroalkyl oligosaccharide had an EC ₅₀ of 0.19 to 0.57 μg / ml, which was extremely high activity against HIV. Further, in all cases, CC ₅₀ has low toxicity of 1000 μg / ml or more, and the sulfated fluoroalkyl oligosaccharide of the present invention has high activity against HIV,
It was revealed to have low toxicity.

The present invention selects a compound excellent as an antiviral agent and an intermediate thereof, but a compound useful as an antiviral agent and an intermediate thereof even if the sugar moiety is other than laminaripentaose. There is. The sulfated fluoroalkyl oligosaccharides or peracetylfluoroalkyl oligosaccharides will be described. As sugars constituting the sugar moiety, there are 6-carbon sugars such as glucose, galactose, and mannose, and other commonly known monosaccharides. As the oligosaccharide moiety composed of these monosaccharides, all oligosaccharides having two or more sugars such as homooligosaccharides, heterooligosaccharides and branched oligosaccharides can be used.

The number of monosaccharides constituting the oligosaccharide moiety of the fluoroalkyl oligosaccharide may be any number, but generally, the synthesis yield decreases as the number of sugar chains increases, so the oligosaccharide moiety is preferably 30 or less. From the viewpoint of antiviral activity, the number of monosaccharides varies depending on the type of sugar used, but is usually preferably 30 or less in view of the anticoagulant action and the compatibility with living organisms which are observed in the case of sulfated polysaccharides. Regarding the oligosaccharide chain length, the activity changes depending on the type of sugar,
Glucose-based oligosaccharides give good performance with sugar chains of 3 or more sugars, and more preferably give extremely good results when the sugar chain has about 5 sugar chains. The glycosidic bond between monosaccharides is
Any of (1 → 2), (1 → 3), (1 → 4), and (1 → 6) may be used, but α (1 → 4) and α (1 → 6) -linked sulfated fluoroalkyl oligosaccharides are , Because it is easily decomposed in vivo, when it is used as an antiviral agent, it easily decomposes in the body and loses its activity. Will be required. Therefore, the oligosaccharide moiety of the sulfated fluoroalkyl oligosaccharide is more preferably a β-bond having a relatively smaller biodegradability than that of an α-bond, and the bonding position is also β (1 → 4), β (1 → 6) more than 6)
A (1 → 3) bond is more preferable. More specifically, β (1 → 3) oligoglucose (an oligosaccharide in which glucose is sequentially bound at the 1st and 3rd positions of glucose), that is, a laminari oligosaccharide obtained by decomposing a polysaccharide such as curdlan or laminaran is preferable.

Orient constituting fluoroalkyl oligosaccharide
More specific examples of goose are shown below. In addition, sugar water
For the acid group, describe the underivatized oligosaccharide
However, the sulphate group or acetyl group
Of the fluoroalkyl group at the reducing sugar 1-position.
Those that are combined are useful as antiviral agents. D-glucopyranosyl- (1 → 3)-{β-D-gluco
Pyranosyl (1 → 3)} _n-Β-D-glucopyrano
, D-glucopyranosyl- (1 → 4)-{α-D-glucose
Pyranosyl- (1 → 4)}_n-Α-D-glucopyrase D-glucopyranosyl- (1 → 6)-{β-D-glucose
Pyranosyl- (1 → 6)}_n-Β-D-glucopyrase D-galactopyranosyl- (1 → 4)-{β-D-gala
Topyranosyl- (1 → 4)}_n-Β-D-glucopyra
Note that n represents an integer of 0 to 28.

[0037]

EXAMPLES The present invention will be described in more detail with reference to the examples according to the present invention.

Example 1 Synthesis of 1H, 1H, 2H, 2H-perfluorodecyl-β-D-laminaripentaoside peracetate 0.30 g of D-laminaripentaose superacetate (JP-A-5-78382) Synthetic for reference) and 0.190 g
1H, 1H, 2H, 2H-perfluorodecanol was dissolved in 10 ml of hexafluoro-meta-xylene, an appropriate amount of molecular sieve was added, and the temperature was adjusted to 4
Add 0.102 g of tin tetrachloride while stirring at 5 ° C,
It was stirred for 2 hours. The reaction mixture was diluted with chloroform and washed with a saturated aqueous sodium hydrogen carbonate solution, water and saturated brine in this order. Further purification by column chromatography (silica gel / hexane / ethyl acetate) and analysis by silica gel thin layer chromatography (ethyl acetate / hexane / = 3/1 (volume ratio)) collects and concentrates a fraction having an Rf value of 0.61. did. The yield was 0.079 g and the yield was 21%.
This compound is 1 from the nuclear magnetic resonance spectrum shown below.
It was confirmed to be H, 1H, 2H, 2H-perfluorodecylperacetyl-β-D-laminaripentaoside.

Nuclear magnetic resonance spectrum (proton) 1-position 4.52 ppm of ring proton of sugar bound to alcohol, J12 = 8.5 Hz, 1 proton Carbon-proton of 2-position of fluoroalkyl 2.30-2.53 ppm , Multiplet, 2 protons Specific rotation [α] D = −43.8 ° (c = 1.0 / chloroform) (20 ° C.)

Example 2 Sulfation 1H, 1H, 2H, 2
Synthesis of H-perfluorodecyl-β-D-laminaripentaoside 70 mg of the fluoroalcohol glycoside peracetate obtained in Example 1 was dissolved in 5 ml of methanol to obtain 3.7.
After adding 0.1 ml of a 0.1 M sodium methoxide methanol solution at room temperature and stirring for 5 hours, sodium ions were removed by an ion exchange resin. After filtering off the ion exchange resin, methanol was concentrated to dryness, and 45 mg of 1H, 1H, 2
H, 2H-perfluorodecyl-β-D-laminaripentaoside was obtained. 4 of the obtained 1H, 1H, 2H, 2H-perfluorodecyl-β-D-laminaripentaoside
5 mg was dissolved in 5 ml dry pyridine by heating to 85 ° C. and 176 mg sulfur trioxide pyridine complex was added,
The mixture was stirred at the same temperature for 90 minutes. After cooling to room temperature, dilute with 3 ml of distilled water, adjust the pH to 7-8 with barium hydroxide,
The solid matter was separated by centrifugation, passed through a sodium ion type ion exchange resin column to give a sodium salt, and the distillate was concentrated under reduced pressure. The residue is very small (0.5m
It was dissolved in distilled water (about 1 l) and reprecipitated with 300 ml of acetone. The resulting precipitate was collected by centrifugation, washed with acetone three times, and freeze-dried from water to obtain the desired product. Yield 68 mg.

Specific optical rotation [α] D = -11.9 ° (c =
1.0 / water) (20 ° C.) Elemental analysis value C 14.2 wt%, H 2.8 wt%,
S 15.9 wt%, Na 11.6 wt%

Example 3 Synthesis of 1H, 1H, 2H, 2H-perfluorododecyl-β-D-laminaripentaoside peracetate 0.20 g D-laminaripentaose superacetate and 0.250 g 1H, 1H, 2H, 2H-perfluorododecanol was dissolved in 10 ml of methylene chloride, and after adding an appropriate amount of molecular sieve, 0.068 g of tin tetrachloride was added under reflux and refluxed for 2.5 hours. It was The reaction mixture was diluted with chloroform and washed with saturated aqueous sodium hydrogen carbonate solution, water and saturated brine. Further purification by column chromatography (silica gel / hexane / ethyl acetate) and analysis by silica gel thin layer chromatography (ethyl acetate / hexane / = 3/1 (volume ratio)) collects and concentrates a fraction having an Rf value of 0.68. did. The yield was 0.058 g and the yield was 22%. This compound is 1H, 1H, 2H, 2 from the nuclear magnetic resonance spectrum shown below.
It was confirmed to be H-perfluorododecyl peracetyl-β-D-laminaripentaoside.

Nuclear magnetic resonance spectrum (proton) 1st position of ring proton of sugar bound to alcohol 4.50 ppm, J12 = 7.8 Hz, 1 proton Carbon proton of 2nd position of fluoroalkyl 2.30-2.55 ppm , Multiplet, 2 protons Specific rotation [α] D = -42.1 ° (c = 1.0 / chloroform) (20 ° C.)

Example 4 Sulfation 1H, 1H, 2H, 2
Synthesis of H-perfluorododecyl-β-D-laminaripentaoside 65 mg of the fluoroalcohol glycoside peracetate synthesized according to Example 3 is dissolved in 5 ml of methanol and 3.2 ml of 0.1N sodium methoxide methanol solution is dissolved. Was added dropwise at room temperature and stirred for 5 hours, and then sodium ions were removed by an ion exchange resin. After removing the ion exchange resin by filtration, the methanol was concentrated to dryness to obtain 35 mg of 1H, 1.
H, 2H, 2H-perfluorododecyl-β-D-laminaripentaoside was obtained. The obtained 1H, 1H, 2H,
35 mg of 2H-perfluorododecyl-β-D-laminaripentaoside in 85 ml of dry pyridine was
155 mg of sulfur trioxide pyridine complex was added, and the mixture was stirred at the same temperature for 90 minutes. After cooling to room temperature, dilute with 3 ml of distilled water and adjust the pH to 7 with barium hydroxide.
The solid content was separated by centrifugation, passed through a sodium ion type ion exchange resin column to give a sodium salt, and the distillate was concentrated under reduced pressure. The residue was dissolved in a very small amount (about 0.3 ml) of distilled water and reprecipitated with 250 ml of acetone. The resulting precipitate was collected by centrifugation, washed with acetone three times, and freeze-dried from water to obtain the desired product. Yield 3
9 mg.

Specific rotation [α] D = −9.8 ° (c =
1.0 / water) (20 ° C.) Elemental analysis value C 13.7 wt%, H 2.6 wt%,
S 15.5 wt%, Na 10.8 wt%

Example 5 1H, 1H, 2H, 2H, 3
Synthesis of H, 3H, 4H, 4H, 5H, 5H, 6H, 6H-perfluorodecyl-β-D-laminaripentaoside peracetate 0.30 g D-laminaripentaose superacetate and 0.124 g 1H, 1H, 2H, 2H, 3H, 3
H, 4H, 4H, 5H, 5H, 6H, 6H-perfluorodecanol dissolved in 10 ml of methylene chloride,
After adding an appropriate amount of molecular sieves, 0.102 g of tin tetrachloride was added under reflux, and the mixture was refluxed for 2.5 hours.
The reaction mixture was diluted with chloroform and washed with saturated aqueous sodium hydrogen carbonate solution, water and saturated brine. Further purification by column chromatography (silica gel / hexane / ethyl acetate) and analysis by silica gel thin layer chromatography (ethyl acetate / hexane / = 3/1 (volume ratio))
The Rf value of 0.71 was collected and concentrated. Yield 0.11
6 g, yield 33%. From the nuclear magnetic resonance spectrum shown below, this compound is H, 1H, 2H, 2H, 3H,
It was confirmed to be 3H, 4H, 4H, 5H, 5H, 6H, 6H-perfluorodecyl-β-D-laminaripentaoside peracetate.

Nuclear magnetic resonance spectrum (proton) 1-position of ring proton of sugar bound to alcohol 4.50 ppm, J12 = 8.4 Hz, 1 proton Proton of 6-position of fluoroalkyl group 2.08 ppm Multiplet line, 2 Proton Specific rotatory power [α] D = -44.1 ° (c = 1.0 / chloroform) (20 ° C.)

Example 6 Sulfation 1H, 1H, 2H, 2
H, 3H, 3H, 4H, 4H, 5H, 5H, 6H, 6H
-Synthesis of perfluorodecyl-β-D-laminaripentaoside 83 mg of the fluoroalcohol glycoside peracetate obtained in Example 5 was dissolved in 5 ml of methanol to obtain 3.2.
After adding 0.1 ml of a 0.1 M sodium methoxide methanol solution at room temperature and stirring for 5 hours, sodium ions were removed by an ion exchange resin. After the ion exchange resin was filtered off, methanol was concentrated to dryness to obtain 51 mg of 1H, 1H, 2
H, 2H, 3H, 3H, 4H, 4H, 5H, 5H, 6
H, 6H-perfluorodecyl-β-D-laminaripentaoside was obtained. 5 of the obtained 1H, 1H, 2H, 2H-perfluorodecyl-β-D-laminaripentaoside
1 mg was dissolved in 5 ml of dry pyridine by heating to 85 ° C., 225 mg of sulfur trioxide pyridine complex was added,
The mixture was stirred at the same temperature for 90 minutes. After cooling to room temperature, dilute with 3 ml of distilled water, adjust the pH to 7-8 with barium hydroxide,
The solid matter was separated by centrifugation, passed through a sodium ion type ion exchange resin column to give a sodium salt, and the distillate was concentrated under reduced pressure. The residue is very small (0.3m
It was dissolved in distilled water (about 1 l) and reprecipitated with 250 ml of acetone. The resulting precipitate was collected by centrifugation, washed with acetone three times, and freeze-dried from water to obtain the desired product. Yield 95 mg.

Specific Optical Rotation [α] D = −9.6 ° (c =
1.0 / water) (20 ° C.) Elemental analysis value C 16.4 wt%, H 2.7 wt%,
S 16.4 wt%, Na 11.7 wt%

Example 7 1H, 1H, 2H, 2H, 3
Synthesis of H, 3H, 4H, 4H, 5H, 5H, 6H, 6H-perfluorotetradecyl-β-D-laminaripentaside peracetate 0.30 g D-laminaripentaose superacetate and 0.202 g 1H, 1H, 2H, 2H, 3H, 3
H, 4H, 4H, 5H, 5H, 6H, 6H-perfluorotetradecanol was dissolved in 10 ml of methylene chloride, an appropriate amount of molecular sieve was added, and then 0.102 g of tin tetrachloride was added under reflux. The mixture was refluxed for 2.5 hours. The reaction mixture was diluted with chloroform and washed with saturated aqueous sodium hydrogen carbonate solution, water and saturated brine. Further purification by column chromatography (silica gel / hexane / ethyl acetate) and analysis by silica gel thin layer chromatography (ethyl acetate / hexane / = 3/1 (volume ratio)) collects and concentrates a fraction having an Rf value of 0.71. did. The yield was 0.129 g and the yield was 33%. This compound shows H, 1H, 2H, 2 from the nuclear magnetic resonance spectrum shown below.
H, 3H, 3H, 4H, 4H, 5H, 5H, 6H, 6H
-Perfluorotetradecyl-β-D-laminaripentaoside peracetate was confirmed.

Nuclear magnetic resonance spectrum (proton) 1-position of ring proton of sugar bound to alcohol 4.50 ppm, J12 = 8.4 Hz, 1-proton Proton of 6-position of fluoroalkyl group 2.08 ppm Multiplet, 2 Proton Specific rotation [α] D = -48.6 ° (c = 1.0 / chloroform) (20 ° C)

Example 8 Sulfation 1H, 1H, 2H, 2
H, 3H, 3H, 4H, 4H, 5H, 5H, 6H, 6H
-Synthesis of perfluorotetradecyl-β-D-laminaripentaoside 98 mg of the fluoroalcohol glycoside peracetate obtained in Example 7 was dissolved in 5 ml of methanol to give 3.8.
After adding 0.1 ml of a 0.1 M sodium methoxide methanol solution at room temperature and stirring for 5 hours, sodium ions were removed by an ion exchange resin. After the ion exchange resin was filtered off, methanol was concentrated to dryness to obtain 54 mg of 1H, 1H, 2
H, 2H, 3H, 3H, 4H, 4H, 5H, 5H, 6
H, 6H-perfluorotetradecyl-β-D-laminaripentaoside was obtained.

54 mg of the obtained 1H, 1H, 2H, 2H-perfluorotetradecyl-β-D-laminaripentaoside was heated to 85 ° C. in 5 ml of dried pyridine and dissolved to give 238 mg of sulfur trioxide. A pyridine complex was added, and the mixture was stirred at the same temperature for 90 minutes. After cooling to room temperature, the mixture was diluted with distilled water (3 ml), the pH was adjusted to 7 to 8 with barium hydroxide, the solid substance was separated by centrifugation, passed through a sodium ion type ion exchange resin column to give a sodium salt, and the distillate was removed. Concentrated under reduced pressure. The residue was dissolved in a very small amount (about 0.3 ml) of distilled water and reprecipitated with 250 ml of acetone. The resulting precipitate was collected by centrifugation, washed with acetone three times, and freeze-dried from water to obtain the desired product. Yield 9
6 mg.

Specific optical rotation [α] D = −9.6 ° (c =
1.0 / water) (20 ° C.) Elemental analysis value C 17.3 wt%, H 2.6 wt%,
S 15.1 wt%, Na 10.8 wt%

(Test Example) Measurement example of anti-HIV activity Test method: MTT method Cell: MT-4 cell Virus: HTLV-IIIB (infectious titer: 3 × 10 5 PF)
U / ml) The outline of the test is shown below.

HIV-infected MT-4 cells (2.
5x104 / well, MOI: 0.01) was added immediately after infection. In order to know the cytotoxicity of the test substances against MT-4, virus-uninfected cells were likewise incubated with different concentrations of the test substances. 37 in a CO ₂ incubator
After culturing at 5 ° C for 5 days, MTT (3- (4,5-dim
ethylthiazol-2-yl) -2,5-di
phenyl tetrazolim bromid
e) PBS (-) aqueous solution (7.5 mg / ml) was injected, and after incubation, IPA / HCl / TritonX
The OD value of formazan precipitated only in living cells by adding -100 solution was measured, and the number of viable cells was measured. From this value, the concentration that protects against cell damage caused by HIV infection by 50% (EC ₅₀ ; 50% effective c)
oncentration) and cytotoxicity are 50% cytotoxicity (CC ₅₀ ; 50% cyto) of the test substance.
Toxic concentration was calculated to obtain anti-HIV activity data (Powell et al.
Auwels, et al) Journal Biological Method, Vol. 20 (1988) 309-321). The compounds described in Examples 2, 4, 6, and 8 were used as test substances. The results are shown in Table 1.

[0057]

[Table 1] Table 1 ─────────────────────────
── Sample CC ₅₀ EC ₅₀ ──────────────────────────
Example 2> 1000 0.57 Example 3> 1000 0.19 Example 4> 1000 0.70 Example 8 945 0.53 Comparative Example 1 ¹⁾ > 1000 0.61 Comparative Example 2 ²⁾ > 1000 0.18 ─────────────────────────
── 1) Sulfated n-dodecyl laminaripentaoside 2) Curdlan sulfate (molecular weight)

The following are examples of drug production of the antiviral agent of the present invention. (Manufacture of Drug 1) Sulfated fluoroalkyl laminaripentaoside (Compound of Example 4) 50 mg Starch 30 mg Lactose 110 mg Talc 7 mg Magnesium stearate 3 mg ────────────────── ─────────────────── 200 mg Sulfated 1H, 1H, 2H, 2H-perfluorododecyl-β-D-laminaripentaoside (Compound of Example 4) Crush, add lactose and starch to it, and mix. 10% starch paste is added to the above mixture and stirred to produce granules. After drying, the particles are sized, mixed with talc and magnesium stearate, and compressed into tablets by a conventional method.
00 mg tablets were produced.

(Drug Production Example 2) Sulfated fluoroalkyl laminaripentaoside (compound of Example 4) 25 mg Starch 23 mg Lactose 50 mg Magnesium stearate 2 mg ──────────────── ──────────────────── 100 mg Sulfated 1H, 1H, 2H, 2H-perfluorododecyl-β-D-laminaripentaoside (Compound of Example 4 ) 25 mg was crushed well, lactose, starch and magnesium stearate were added, mixed well and then filled into capsules to prepare capsules.

(Drug Production Example 3) Sulfation 1H, 1H, 2
To 400 mg of H, 2H-perfluorododecyl-β-D-laminaripentaoside (compound of Example 4), physiological saline sterilized by an autoclave was added and dissolved to bring the total amount to 1
After adjusted to 0 ml, the solution was sealed in a dry heat sterilized ampoule bottle to prepare 10 ml of a liquid preparation.

[0061]

INDUSTRIAL APPLICABILITY The present invention provides a novel sulfated fluoroalkyl laminaripentaoside and peracetylfluoro laminaripentaoside, and particularly sulfated fluoroalkyl laminaripentaoside and its physiologically acceptable salts. Is a highly lipophilic compound having a fluorine atom in the alkyl group, particularly HIV.
It is possible to provide an antiviral agent having an excellent action against

Claims (9)

[Claims] 1. A hydrogen atom of a hydroxyl group at the 1-position of the reducing terminal sugar of laminaripentaose is substituted with a fluoroalkyl group,
A sulfated fluoroalkyl laminaripentaoside or a physiologically acceptable salt thereof in which a part or all of the remaining hydroxyl groups are substituted with a sulfate ester group. 2. The fluoroalkyl group has 10 to 14 carbon atoms.
2. A sulfated fluoroalkyl laminaripentaoside or a physiologically acceptable salt thereof according to claim 1, which is a fluoroalkyl group of 3. A fluoroalkyl group has the formula: --C ₂ H ₄ C ₈ F ₁₇ , --C ₂ H ₄ C ₁₀ F ₂₁ , --C ₆ H.
The sulfated fluoroalkyl laminaripentaoside or a physiologically acceptable salt thereof according to claim 1, which is a fluoroalkyl group selected from the group consisting of ₁₂ C ₄ F ₉ or —C ₆ H ₁₂ C ₈ F _17. salt. 4. The sulfated fluoroalkyl laminaripentaoside or the physiology thereof according to claim 3, wherein the fluoroalkyl group is a fluoroalkyl group represented by: —C ₂ H ₄ C ₁₀ F ₂₁ . Acceptable salt. 5. The hydrogen atom of the hydroxyl group at the 1-position of the reducing terminal sugar of laminaripentaose is substituted with a fluoroalkyl group,
And a peracetylfluoroalkyl laminaripentaoside in which all the remaining hydroxyl groups are substituted with acetyl groups. 6. The fluoroalkyl group has 10 to 14 carbon atoms.
The peracetylfluoroalkyl laminaripentaoside according to claim 1, which is a fluoroalkyl group of 7. A fluoroalkyl group, embedded image _{-C 2 H 4 C 8 F 17} , -C 2 H 4 C 10 F 21, -C 6 H
_The peracetylfluoroalkyl laminaripentaoside according to claim 5, which is a fluoroalkyl group selected from the group consisting of ₁₂ C ₄ F ₉ or —C ₆ H ₁₂ C ₈ F ₁₇ . 8. An antiviral agent comprising the sulfated fluoroalkyl laminaripentaoside according to any one of claims 1 to 4 as an active ingredient. 9. The antiviral agent according to claim 8, wherein the virus is HIV.