JPH04227601A - Polysulfate of cyclodextrin sulfide derivative and its production - Google Patents

Abstract

PURPOSE:To improve an antiretrovirus function by reacting a specific sulfodextrin sulfide derivative with a sulfonating agent. CONSTITUTION:A cyclodextrin sulfide derivative in which at least one of 6 to 8 D-glucose units constituting the cyclodextrin is a unit of the formula (wherein R' is an alkyl which may be substituted by 1 to 3 substituted or unsubstituted phenyl groups, a phenyl which may have a substituent, or a nitrogenous heterocyclic group which may have a substituent) is allowed to react with 1 to 10 equivalents of a sulfonating agent based on the hydroxyl groups of the derivative under heating in a reaction solvent.

Description

[Detailed description of the invention]

0001

FIELD OF THE INVENTION The present invention relates to a novel polysulfate ester compound of a cyclodextrin sulfide derivative having antiretroviral activity and a method for producing the same.

0002

[Prior Art] AIDS (acquired immunodeficiency syndrome) is caused by the human immunodeficiency virus (HIV), a type of retrovirus.
) is a fatal or extremely malignant disease caused by infection, and its prevention and eradication is currently the most important problem for humanity to overcome on a global level. Conventionally, as a compound having an antiretroviral effect, for example, azidothymidine [Igaku no Ayumi, Vol. 142, No. 9, 619
-622 pages (1987)] and sulfated polysaccharides (JP-A-63-45223, JP-A-64-25724). However, whether conventionally known antiretroviral drugs are effective and safe for the treatment of AIDS has not yet been fully verified and confirmed.

0003

OBJECTS OF THE INVENTION An object of the present invention is to provide a novel pharmaceutical compound having excellent antiretroviral activity, particularly excellent antiproliferative activity against HIV.

0004

[Means for Solving the Problems] The present invention provides that at least one of the 6 to 8 D-glucose units constituting a cyclodextrin has the general formula (I).

[0005]

[Chemical formula 3]

(However, R1 is an alkyl group that may be substituted with 1 to 3 substituted or unsubstituted phenyl groups, a phenyl group that may have a substituent, or a phenyl group that may have a substituent. This invention relates to a polysulfate ester of a cyclodextrin derivative (hereinafter referred to as a polysulfate compound) or a salt thereof, which is a unit represented by a nitrogen-containing heterocyclic group.

The polysulfate ester compound of the present invention is schematically shown as follows.

[0008]

[C4]

(However, n is an integer from 6 to 8, m is an integer from 1 to n, and R is -SO3H in whole or in part.
and the remainder are hydrogen atoms, and R1 has the same meaning as above. ) The above general formula (II) represents that 6 to 8 structural units are cyclically bonded at the 1st and 4th positions in any order. Further, in general formula (II), n is 6
A compound where n is 7 is a derivative of α-cyclodextrin, a compound where n is 7 is a derivative of β-cyclodextrin, and a compound where n is 8 is a derivative of γ-cyclodextrin.

Specific examples of the polysulfate ester compound of the present invention include, in the above general formula (II), R1 is a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group,
an alkyl group having 1 to 20 carbon atoms such as an octadecyl group;
a lower alkyl group substituted with 1 to 3 phenyl groups, halogenophenyl groups or lower alkoxyphenyl groups, such as a benzyl group, trityl group, phenethyl group, chlorobenzyl group or methoxybenzyl group; (1) phenyl group, chlorophenyl group; A phenyl group which may be substituted with a halogen atom, a lower alkyl group or a lower alkoxy group, such as a methylphenyl group or a methoxyphenyl group, and a phenyl group which is substituted with 1 to 2 hydroxyl groups, such as a dihydroxypyrimidinyl group or a purinyl group. Examples include compounds which are optionally a pyrimidinyl group or a purinyl group.

The above polysulfate ester compound (I) of the present invention
Among I), preferred compounds include, for example, R1
is a benzyl group, a chlorobenzyl group, a methoxybenzyl group, a methylphenyl group, a methoxyphenyl group, or a purinyl group. Further, the number of sulfate groups contained in the polysulfate compound (II) of the present invention is preferably 8 to 23. In the present invention, the lower alkyl group and the lower alkoxy group include those having 1 to 6 carbon atoms, particularly those having 1 to 4 carbon atoms.

Polysulfate compound (II) of the present invention
is produced by reacting a β-cyclodextrin derivative in which at least one of the 6 to 8 D-glucose units constituting the cyclodextrin is a unit represented by the general formula (I) and a sulfonating agent. can do. The above reaction is schematically shown as follows.

[0013]

[C5]

(However, the symbols have the same meanings as above.)
The reaction between compound (III) and the sulfonating agent can be carried out in a suitable solvent. As the sulfonating agent, for example, sulfur trioxide-pyridine complex, sulfur trioxide-trialkylamine complex, sulfuric anhydride, concentrated sulfuric acid, chlorosulfuric acid, etc. can be used. Further, it is preferable to use the sulfonating agent in an excess amount relative to the raw material compound (III). For example, when using a sulfur trioxide-pyridine complex or a sulfur trioxide-trialkylamine complex as the sulfonating agent, is used in an amount of about 1 to 10 equivalents, preferably about 2 to 5 equivalents, based on the hydroxyl group of the starting compound (III).

As the reaction solvent, for example, tertiary amine (
For example, pyridine, picoline, lutidine, N,N-dimethylaniline), N,N-dimethylformamide, hexamethylenephosphorylamide, dimethylsulfoxide,
Chloroform, benzene, toluene, xylene, water, a mixture thereof, liquid sulfite, and the like can be suitably used. This reaction can be carried out under cooling to heating, preferably under heating. After the reaction is complete,
The target product can be isolated and purified by conventional methods. For example, the target product can be obtained as an alkali metal salt by treating the crude product obtained from the reaction-completed liquid with an alkali metal hydroxide and then applying it to a column packed with a crosslinked dextran gel or the like.

The polysulfate ester compound of the present invention can be suitably used either in its free form or in its pharmacologically acceptable salt form. Such salts include, for example, alkali metal salts such as sodium salts, potassium salts, lithium salts, calcium salts, magnesium salts,
Examples include alkaline earth metal salts such as barium salts, organic amine salts such as trimethylamine salts, triethylamine salts, pyridine salts, and glycine ethyl ester salts. The polysulfate compound or salt thereof of the present invention can be administered orally or parenterally (e.g., intravenously, intramuscularly, subcutaneously), and can be administered in the form of tablets, granules, capsules, powders, etc. in a conventional manner. It can be used in appropriate pharmaceutical formulations such as injections, suppositories, vaginal preparations, and creams. Although the dosage of the active ingredient compound of the present invention varies depending on the patient's age, weight, condition, and type of disease, it is usually about 0.5 mg/day.
1 to 500 mg/kg is suitable, especially about 1 to 50 m
It is preferable to set it to about g/kg.

[0017]

[Example] Example 1 heptakis(6-benzylthio-6-deoxy)-β-
Add 0.4 g of cyclodextrin to 25 ml of pyridine, then add 1.4 g of sulfur trioxide-pyridine complex.
Add 2g and stir at 70°C for 6 hours. After cooling, the supernatant liquid was removed, and the residue was dried under reduced pressure, then dissolved in 5 ml of water, adjusted to pH 8 with 10% sodium hydroxide, and then applied to a column packed with Sephadex G-10 (manufactured by Pharmacia). By purifying with
get. IRNujolνMaxcm-1:1240,
1160,1040,8301H-NMR(D2O)δ
: 3.6 (br, s), 6.8 to 7.4 (br, s) Number of sulfate ester groups in the molecule calculated from elemental analysis values:
12

Examples 2 to 19 The corresponding raw material compounds were treated in the same manner as in Example 1, and the following
The compounds listed in the table are obtained.

[0019]

[Table 1]

[0020]

[Table 2]

[0021]

[Table 3]

[0022]

[Table 4]

Examples 20 to 23 Tris(6-benzylthio-6-deoxy)-γ-cyclodextrin or octakis(6-deoxy) obtained in Reference Example 22
Benzylthio-6-deoxy)-γ-cyclodextrin is treated in the same manner as in Example 1 to obtain the compounds listed in Table 2 below.

[0024]

[Table 5]

Example 24 Hexakis(6-p-tolylthio-6-deoxy)-α
-Cyclodextrin was treated in the same manner as in Example 1 to produce hexakis(6-p-tolylthio-6-deoxy)-α-
Obtain the sodium salt of cyclodextrin polysulfate. Yield: 126% (weight % of target product relative to starting compound) IRNujolνMaxcm-1: 1240,
1040,8301H-NMR (D2O) δ: 2.0(
brs), 6.5-7.2 (brm) Number of sulfate ester groups in the molecule calculated from elemental analysis value: 10

0026
Reference Example 1 Add 30 ml of mesitylenesulfonyl chloride to a 2.5 liter solution of 126 g of β-cyclodextrin in pyridine at 25°C.
Add g little by little. After stirring for 2 hours, 6 g of mesitylenesulfonyl chloride was further added and stirred for 1 hour. After adding water to the reaction solution and leaving it overnight, the solvent was distilled off, and the residue was dissolved in 1 liter of water and applied to a column filled with CHP-20 resin (manufactured by Mitsubishi Chemical Corporation). After sequentially washing the column with water, 10% methanol, and 20% methanol, 50% methanol was passed through the column and the eluate was collected. Next, 80% methanol is passed through the tube and the eluate is collected. The following compounds i) and ii) are obtained by concentrating and drying each eluate under reduced pressure. i) Mono(6-O-mesitylenesulfonyl)-β-cyclodextrin Yield: 52.6g White powder IRNujol νMaxcm-1: 1160,10
80,1030 1H-NMR (DMSO-d6) δ: 2.29 (s, 3
H), 2.54 (s, 6H), 2.7-4.6 (m, 4
8H), 4.6-4.9 (m, 7H), 5.70 (br
s, 14H), 7.10 (s, 2H) ii) Bis(6-O-mesitylenesulfonyl)-β-cyclodextrin Yield: 13.0g White powder IRNujol νMaxcm-1: 1610, 13
55,1160,1030 1H-NMR (DMSO-d6) δ: 2.29 (s, 6
H), 2.52 (s, 12H), 3.1-4.6 (m,
47H), 4.74(brs, 4H), 4.84(br
s, 3H), 5.6-5.9 (m, 14H), 7.07
(s, 2H), 7.10 (s, 2H)

Reference Example 2 56.8 g of β-cyclodextrin and 600 m of pyridine
A solution of 43.7 g of mesitylene sulfonyl chloride in 100 ml of pyridine was added to the
After the dropwise addition over a period of time, the mixture was further stirred for 2 hours. The solvent is distilled off from the reaction solution, the residue is dissolved in 200 ml of methanol, and 300 ml of water is added. The solution is cooled with ice water, the supernatant is discarded, and the residue is treated with acetone to powder. The powder is collected by filtration, dried, and then separated and purified by silica gel column chromatography to obtain the following compounds i), ii), and iii).

i) Tris(6-O-mesitylenesulfonyl)-β-cyclodextrin yield: 7. Og White powder IRKBr νMaxcm-1:3400,2850
, 1610, 1350, 1190, 1175, 1155
, 1080, 1030 1H-NMR (DMSO-d6) δ: 2.2-2.4 (
m, 9H), 2.4-2.7 (m, 18H), 3.0-
5.0 (br, 53H), 5.6-6.0 (m, 14H
), 7.0-7.3 (m, 6H) ii) Tetrakis(6-O-mesitylenesulfonyl)-
β-Cyclodextrin yield: 3.0g white powder IRKBr νMaxcm-1: 3400,2850
,1610,1355,1190,1170,1080
, 1055 1H-NMR (DMSO-d6) δ: 2.27 (br,
s, 12H), 2.4-2.7 (m, 24H), 3.0
~5.0 (m, 52H), 5.6 ~ 6.1 (m, 14H
), 6.9-7.2 (m, 8H) iii) Pentakis (6-O-mesitylenesulfonyl)
-β-Cyclodextrin yield: 1.30g White powder IRKBr νMaxcm-1: 3400, 2850
,1610,1355,1190,1175,1080
, 1055 1H-NMR (DMSO-d6) δ: 2.26 (brs
, 15H), 2.3~2.7 (m, 30H), 3.0~
5.0 (m, 51H), 5.6-6.1 (m, 14H)
, 6.8-7.2 (m, 10H)

Reference Example 3 To a solution of 0.69 g of benzyl mercaptan in 20 ml of dimethylformamide, 0.22 g of sodium hydride (62
% content) while stirring while cooling with ice water. Heptakis(6-iodo-6-deoxy)-β was added to the mixture at room temperature.
- Add 1 g of cyclodextrin, under a stream of argon gas,
Incubate overnight at room temperature. By pouring the reaction solution into 200 ml of water, filtering the resulting precipitate, washing and drying,
heptakis(6-benzylthio-6-deoxy)-β-
0.95 g of yellow powder of cyclodextrin is obtained. 1H-NMR (DMSO-d6) δ: 4.9 (brs)
, 5.82 (brs), 7.17 (brs)

0030
Reference Example 4 Benzyl mercaptan 3 was added to a suspension of 1.2 g (62%) of sodium hydride in 130 ml of dimethylformamide.
．． 5 ml of mono(6-O) was added dropwise to the mixture after 30 minutes.
-mesitylenesulfonyl)-β-cyclodextrin 1
Add 3.2g and stir at 80°C for 8 hours. After the reaction solution was cooled, it was poured into 600 ml of acetone, and the precipitate was collected by filtration and dried to obtain a colorless powder of mono(6-benzylthio-6-deoxy)-β-cyclodextrin (9.84 g).
get g. 1H-NMR (DMSO-d6) δ: 4.8
3(brs), 5.73(brs), 7.2~7.4(
m)

Reference Examples 5 to 21 The corresponding starting compounds were treated in the same manner as in Reference Example 4 to obtain the compounds listed in Table 3 below.

[0032]

[Table 6]

[0033]

[Table 7]

[0034]

[Table 8]

[0035]

[Table 9]

Reference Example 22 (1) 39.1 g of γ-cyclodextrin was added to pyridine 6
00 ml solution, and add 26 g of mesitylenesulfonyl chloride while stirring at room temperature. After stirring at room temperature overnight, water was added to the reaction solution and the solvent was distilled off. The residue is washed with water and ethanol, filtered as a powder, then dissolved in methanol and applied to a column packed with CHP-20 resin (manufactured by Mitsubishi Kasei Corporation). The column was washed with 50% methanol, 6
After sequentially washing with 5% methanol, 80% methanol is passed through to elute tris(6-O-mesitylenesulfonyl)-γ-cyclodextrin. Next, methanol was passed through, and tetrakis(6-O-mesitylenesulfonyl)-
Elute γ-cyclodextrin.

(2) 80% methanol eluate [Tris(
[containing 6-O-mesitylenesulfonyl)-γ-cyclodextrin] was concentrated to dryness under reduced pressure, further dissolved in methanol, and then applied to a column packed with CHP-20 resin. After washing the column with 70% methanol, 80% methanol is passed through the column to obtain three fractions (fractions A, B, and C). The HPLC analysis results (retention time) of each fraction are as follows. Fraction A: 7.4, 9.0, 9.5, 11.5 (
Fraction B: 11.6 (min) Fraction C: 7.4 (min) By concentrating and drying each fraction under reduced pressure, the following tris(6-O-mesitylenesulfonyl)-γ- The substituted isomer of cyclodextrin is obtained as a colorless powder.

i) Tris(6-O-mesitylenesulfonyl)-γ-cyclodextrin [prepared from fraction A] Yield: 5.0 g m.p. p. 188°C (decomposition) ii) Tris(6-O-mesitylenesulfonyl)-
γ-Cyclodextrin [prepared from fraction B] Yield: 1.12 g m. P. 192°C (decomposition) iii) Tris(6-O-mesitylenesulfonyl)
-γ-cyclodextrin [prepared from fraction C]
Yield: 1.20g m. P. 190°C (decomposition) On the other hand, the methanol eluate [containing tetrakis(6-O-mesitylenesulfonyl)-γ-cyclodextrin]
Concentrate to dryness under reduced pressure, dissolve 20 g of the obtained residue in 100 ml of pyridine, and add 7.2 g of mesitylenesulfonyl chloride. After stirring at room temperature for 2 days, drying
Octakis (6-O-mesitylenesulfonyl) was purified by silica gel column chromatography.
- gamma-cyclodextrin is obtained as a colorless powder. Yield: 0.93g m. p. 230℃ (decomposition)

(3-a) Benzyl mercaptan 0.
Tris(6-O
-mesitylenesulfonyl)-γ-cyclodextrin 1
．． Add 0g and stir at 80°C for 2 hours. After cooling, add 1 water
The precipitate was collected by filtration, dissolved in methanol, and then decolorized with activated carbon. After concentration, acetone is added, the precipitate is collected by filtration, dissolved in N,N-dimethylformamide, and purified using a column packed with Sephadex G-25 (manufactured by Pharmacia). The eluate is dried, washed with acetone, and dried to obtain 0.28 g of tris(6-benzylthio-6-deoxy)-γ-cyclodextrin as a colorless powder. m. P. 233℃ (decomposition) (3-b~d)
Tris prepared from fractions B and C of (2) (
6-O-mesitylenesulfonyl)-γ-cyclodextrin and octakis(6-O-mesitylenesulfonyl)
-γ-Cyclodextrin is treated in the same manner as in (3-a) to obtain the compounds listed in Table 4 below.

[0040]

[Table 10]

Reference Example 23 0.83 g of P-toluenethiol, 30 ml of N,N-dimethylformamide and 0.25 g of sodium hydride (
1.0 g of hexakis(6-bromo-6-deoxy)-α-cyclodextrin was added to the mixture (containing 62.7%), and the mixture was stirred at room temperature for 20 hours under a stream of argon gas. The reaction solution was poured into 200 ml of water, and the resulting precipitate was collected by filtration, washed, and dried to obtain hexakis(6-p-tolylthio-6-deoxy)-α-cyclodextrin 1.
．． Obtain 0 g as a white powder. m. p. 232-235
°C (decomposition)

[0042]

[Action] Experimental example HIV proliferation inhibitory effect (principle) Human T-cell leukemia virus type I [human T-c
[Ell Leukemia virus, HTLV-I] It is known that when MT-4 cells, a persistently infected cell line, are infected with HIV, HIV rapidly proliferates, and the MT-4 cells die in 5 to 6 days due to cytotoxicity. ing. Therefore, H
The HIV growth inhibitory effect of the sample can be examined using the number of viable MT-4 cells infected with IV as an indicator. (Method) HIV (TALL-1/LAV-1 culture supernatant) was added to MT-4 cells at 0.001TCID50 (median
tissue culture infection
us dose, 50% tissue culture infectious dose)/cell
After infecting for 1 hour at 37°C, the cells were washed and transferred to RPMI-1640 medium [FCS (
fetal calf serum)
containing 10%] at a concentration of 1×10 5 cells/ml. Transfer this cell suspension to a flat bottom culture plate.
00 μl/well and cultured at 37° C. in the presence of 5% carbon dioxide for 5 days. After culturing, the number of living cells in the cell suspension was counted by trypan blue staining. The HIV proliferation inhibitory effect of the specimen was determined as the concentration of the specimen that inhibited 100% of the infectivity and cytopathic effects of HIV on MT-4 cells. (Results) The results are shown in Table 5 below.

[0043]

[Table 11]

[0044]

[Effects of the Invention] As mentioned above, the sulfate ester compound of the present invention has an excellent antiretroviral effect, especially an excellent antiproliferative effect against HIV, and is also characterized by low toxicity and high safety as a medicine. .

Claims (9)

[Claims] Claim 1: 6 to 8 constituting cyclodextrin
At least one of the D-glucose units has the general formula: A polysulfate ester of a cyclodextrin sulfide derivative or a salt thereof. 2. R1 is: (1) an alkyl group having 1 to 20 carbon atoms, (2) a lower alkyl group substituted with 1 to 3 phenyl, halogenophenyl, or lower alkoxyphenyl groups, (2) a halogen atom, or a lower alkyl group; , (2) a phenyl group optionally substituted with a lower alkoxy group, and (1) a pyrimidinyl group or purinyl group optionally substituted with 1 to 2 hydroxyl groups. 3. The compound according to claim 1, wherein R1 is a benzyl group, a chlorobenzyl group, a methoxybenzyl group, a methylphenyl group, a methoxyphenyl group, or a purinyl group. 4. The compound according to claim 1, wherein the cyclodextrin is α-cyclodextrin. 5. The compound according to claim 1, wherein the cyclodextrin is β-cyclodextrin. 6. The compound according to claim 1, wherein the cyclodextrin is γ-cyclodextrin. 7. The compound according to claim 1, having 8 to 23 sulfate groups. Claim 8: 6 to 8 constituting cyclodextrin
At least one of the D-glucose units has the general formula: A cyclodextrin sulfide derivative, which is a unit represented by a phenyl group that may have a substituent or a nitrogen-containing heterocyclic group that may have a substituent, is reacted with a sulfonating agent, and if desired, the product is A method for producing a polysulfate ester of the cyclodextrin sulfide derivative or a salt thereof, characterized in that the polysulfate ester is a salt thereof. 9. The method according to claim 8, wherein the sulfonating agent is sulfur trioxide complex, anhydrous sulfuric acid, concentrated sulfuric acid or chlorosulfuric acid.