JPH04136001A - Polysulfuric ester of sulfonamide derivative of beta-cyclodextrin and preparation thereof - Google Patents

Abstract

PURPOSE:To prepare a medicinal compd. which effectively prevents the growth of a human immunodeficiency virus without showing side effect by reacting a specific sulfonamide deriv. of beta-cyclodextrin with a sulfonating agent. CONSTITUTION:A sulfonamide deriv. of beta-cyclodextrin wherein at least one of the seven D-glucose units constituting beta-cyclodextrin has been substd. by a unit of the formula is reacted with a sulfonating agent, and if necessary the reaction product is converted into a salt, giving a polysulfuric ester of the sulfonamide deriv. or a salt of the ester. In the formula, R<1> is alkyl, phenyl, naphthyl, or heterocyclic. The sulfonamide deriv. is a new compd. and prepd. by reacting beta-cyclodextrin with a substd. sulfonic acid halide, separating and purifying the reaction product if necessary, reacting it with ammonia, and sulfonamidating the resulting product by a suitable method.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to a polysulfate ester of a β-cyclodextrin sulfonamide derivative having antiretroviral activity and a method for producing the same.

(Prior art) AIDS (acquired immunodeficiency syndrome) is a fatal or extremely malignant disease caused by infection with the human immunodeficiency virus (HIV), a type of retrovirus, and prevention and eradication is currently a worldwide goal. It has become the most important problem for humanity to overcome at this level.

Conventionally, compounds with antiretroviral effects include:
For example, azidothymidine [Igaku no Ayumi; Vol. 142, No. 9, pp. 619-622 (1987)], sulfated polyIJi (JP-A No. 63-45223, No. 64-257)
No. 24) etc. are known.

(Problems to be Solved by the Invention) The object of the present invention is to provide excellent antiretroviral effects, especially H.
The purpose of this invention is to provide a new pharmaceutical compound that has an excellent anti-proliferative effect on IV and is free from side effects.

(Structure of the Invention) The present invention provides seven D
- 1 to 7 of the glucose units have the general formula (where RI is an alkyl group; a phenyl group optionally substituted with a lower alkyl group, a lower alkoxy group, or a halogen atom; a naphthyl group; or nitrogen, oxygen, or sulfur) Represents a heterocyclic group containing 1 or 2 heteroatoms selected from and optionally substituted with a lower alkyl group. Polysulfuric acid of a β-cyclodextrin sulfonamide derivative substituted with the unit shown in It relates to an ester (hereinafter referred to as a polysulfate ester compound) or a salt thereof.

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

(However, m is an integer from 1 to 7, and represents that seven structural units are cyclically bonded at the 1st and 4th positions in any order, and R1 to R'' are all or part of -303H1 represents that the remainder is a hydrogen atom, and R1 has the same meaning as above.) As a specific example of the polysulfate ester compound of the present invention, for example, in general formula (I), R1 is a heptyl group, an octyl group, , an alkyl group such as a nonyl group; a lower alkyl group such as a methyl group, an ethyl group, a propyl group, a lower alkoxy group such as a methoxy group, an ethoxy group, a propoxy group, or a halogen atom such as a chlorine atom or a bromine atom. a phenyl group; a naphthyl group; and a heterocyclic group containing one or two atoms selected from nitrogen, oxygen, and sulfur, which may be substituted with a lower alkyl group such as a methyl group, an ethyl group, or a propyl group. (For example, a furyl group, a thienyl group, an oxasilyl group, an isoxasilyl group, a thiazolyl group, an isothiazolyl group).

Among these, compounds in which R1 is a C1-C10 alkyl group; a C1-C4 alkyl group, a C1-4 alkoxy group, or a phenyl group optionally substituted with a halogen atom are medicinally preferable. group; naphthyl group; or a thienyl group, isoxasilyl group, or thiazolyl group optionally substituted with an alkyl group having 1 to 4 carbon atoms, and the number of sulfate ester groups contained in the target compound is 8 to 8.
There are 19 compounds.

The polysulfate ester compound of the present invention comprises one to seven D-glucose units constituting β-cyclodextrin.
It can be produced by reacting a β-cyclodextrin sulfonamide derivative in which seven units are replaced with units represented by the general formula (however, the symbols have the same meanings as above) and a sulfonating agent. .

The above reaction is schematically illustrated as follows.

↓Sulfonating agent (However, the symbols have the same meanings as above.) Sulfonating agents include, for example, sulfur trioxide complex (e.g., sulfur dioxide-pyridine complex, sulfur dioxide-trialkylamine complex, trioxide Sulfur-dioxane complex, sulfur dioxide-N, N-dimethylformamide complex, etc.), sulfuric anhydride, concentrated sulfuric acid, chlorosulfuric acid, etc. can all be suitably used.

Further, the amount of the sulfonating agent used is preferably in excess of the β-cyclodextrin sulfonamide derivative (II). For example, sulfur dioxide-pyridine confusoxol or sulfur trioxide as a sulfonating agent.
When using a trialkylamine complex,
β-cyclodextrin sulfonamide derivative (Il
r) water e! 1 to 10 equivalents, especially 2
It is preferable to use about 5 equivalents.

β-Cyclodextrin sulfonamide derivative (I[
) and a sulfonating agent can be carried out in a suitable solvent or without a solvent. Examples of the reaction solvent include pyridine, N,N-dimethylbormamide, formamide,
hexamethylene phosphoryl triamide, chloroform,
Benzene, toluene, xylene, water and mixtures thereof,
Liquid sulfite or the like can be suitably used.

This reaction can be carried out under cooling or under heating (-30'C to 140''C) depending on the sulfonating agent used.
Particularly when using a sulfur dioxide complex, it is desirable to carry out the process under heating (50'C to 100'C).

After the reaction is complete, if necessary, isolate and isolate the target product using conventional methods.
Can be purified. For example, after neutralizing or salt-exchanging the crude product obtained from the reaction completed liquid with an alkali metal hydroxide, the product is subjected to a column packed with a cross-linked dextran gel or the like, or after being purified with an anion exchange resin, the crude product is purified using an alkali metal hydroxide. By processing with , the target product can be obtained as an alkali metal salt.

The polysulfate ester compound of the present invention can be suitably used either in its free form or in its pharmacologically acceptable salt form. Examples of such salts include alkali metal salts such as sodium salts, potassium salts, and lithium salts; alkaline earth metal salts such as calcium salts, magnesium salts, and barium salts; trimethylamine salts, triethylamine salts, pyridine salts, glycine ethyl ester salts,
Examples include organic amine salts such as ethanolamine salts and basic amino acid salts.

The polysulfate compound of the present invention or a pharmacologically acceptable salt thereof can be administered orally or parenterally (e.g., intravenously, intramuscularly, subcutaneously), and can be administered in the form of tablets, granules, etc. in a conventional manner. It can be used in appropriate pharmaceutical formulations such as tablets, capsules, powders, injections, suppositories, vaginal preparations, and creams.

The dosage of the polysulfate compound of the present invention or its pharmacologically acceptable salt varies depending on the age, weight, condition, and type of disease of the patient, but is usually about 0.1 per day.
~200mg/kg is suitable, especially about 0.5~50mg/kg.
It is preferable to set it to about ng/kg.

Note that the β-cyclodextrin sulfonamide derivative (II), which is the raw material compound of the present invention, is a new! A compound in which β-cyclodextrin is reacted with a sulfonic acid halide having a substituent (for example, mesitylenesulfonyl chloride), and if necessary, after separation and purification, it is reacted with ammonia, and then the product is treated with a sulfonate as appropriate. It can be produced by amidation.

(Example) Example 1 Heptakis (6-benzenesulfonylamino-6-deoxo)-β-cyclodextrin 0. Add 30 g of pyridine to 5 g, then add 1.6 gt of sulfur dioxide pyridine complex, and stir at 70° C. for 6 hours. After the reaction, the supernatant is removed, and the residue is washed with methanol and then ethanol, pulverized, filtered, and dried to obtain a brown powder. This powder was dissolved in three volumes of water, adjusted to pH 8 with 10% sodium hydroxide, and then cross-linked dextran gel:
Purification is performed using a Sephadex G-10 column (manufactured by Pharmacia) to obtain 0.7 g of the sodium salt of heptakis (6-benzenesulfonylamino-6-deoxy)-β-cyclodextrin polysulfate as a white powder.

Yield: 140% (wt% of target product relative to raw material compound,
The same applies to Examples 2 to 14 below) T Rv, , X c
m-': 1240.1)60,1050.830
H-NMR (DzO) 6: 7.0-8.2 (br
s) Number of sulfate ester groups in the molecule calculated from elemental analysis values = 1) Example 2 0.83 g of mono (6-benzenesulfonylamino-6-deoxy)-β-cyclodextrin was dissolved in pyridine 85
- Dissolve by heating at 100°C.

Next, add 6.2 g of sulfur trioxide-pyridine complex without stirring, and stir at 100°C for 6 hours. Pyridine is distilled off, and the residue is dissolved in 20% water and 40% methanol.
-I understand. Furthermore, 300 d of metal vines were added, and the mixture was left standing in a cool place at night. The precipitate is collected by filtration, washed with methanol, dissolved in water, and once concentrated to distill off the methanol contained therein.

Add water and strong acidic ion exchange resin 5K-IB (H゛) to the residue.
Mitsubishi Kasei Corporation) 501) 1 was added and stirred at room temperature for 30 minutes. The resin was filtered off from the mixture, the filtrate was adjusted to pH 7.3 with 017N potassium hydroxide, filtered with acid using a membrane filter, and lyophilized to obtain mono(6-benzenesulfonylamino-6-deoxy)-β-cyclodextrin. Potassium salt of polysulfate ester 2, Og is obtained as a white powder.

Yield: 241% IR, Xc m-': 1540,1240,
1) 60,1000,940゜'If-NMR (DzO
) δ: 7.6 ~ 8. Hm) Number of sulfate ester groups in the molecule calculated from elemental analysis values: 19 Examples 3 to 14 The corresponding raw material compounds were treated in the same manner as in Example 1 or Example 2 to produce the compounds listed in Table 1 below. obtain.

Reference Example 1 1 g of heptakis(6-amino-6-deoxy)-β-cyclodextrin and 4 ml of 10% aqueous sodium hydrogen carbonate solution
0d, then 3.3g of benzenesulfonyl chloride
was added dropwise and stirred vigorously for 2 days at room temperature. The precipitate was collected by filtration, washed with water, and dried to give Hebutakis (6-benzenesulfonylamino 6-deoxy)-β-cyclodextrin 1.
．． 3 g are obtained as a white powder. Yield: 69%'I
I-NMR (DMSO-d6) δ: 4.75 (b
r s), 5.7-5.9 (br s), 7.4-8.
0 (br m) Reference Example 2 (1) 126 g of β-cyclodextrin is dissolved in 2.5j' of pyridine, and 30 g of 2-mesitylenesulfonyl chloride is added in small portions at 25°C. After stirring for 2 hours, 2
- Add 6 g of mesitylenesulfonyl chloride and stir for an additional hour, then add water and leave overnight. After distilling off the solvent from the reaction solution, the residue is dissolved in water 1), and the solution is passed through a column filled with CHP-20 resin (manufactured by Mitsubishi Chemical Corporation). The column is washed sequentially with water, 10% methanol, 20% methanol, and eluted with 50% methanol. The solvent was distilled off from the eluate, and the residue was dried under reduced pressure to obtain 52.6 g of mono(60-mesitylenesulfonyl)-β-cyclodextrin as a white powder.

Yield: 40% I Rv, 41) X cm-': 1) 60,10
80.1030'H-NMR (DMSO-d6) δ
: 2.29(s, 3t(), 2.54(s, 6H)
The column after elution is further eluted with 80% methanol.

The eluate was dried to dryness and dried under reduced pressure to obtain 13.0 g of bis(6-0-mesitylenesulfonyl)-β-cyclodextrin.
is obtained as a white powder.

Yield: 8.7% r RvM, cm-': 1610,1355,
1) 60. IQ30'H-NMR (DMSO-d6)
δ: 2.29 (s, 6H), 2.52 (s, 12H)
(2-a) 26.4 g of mono (6-0-mesitylenesulfonyl)-β-cyclodextrin and 350 m of 10% ammonia-methanol are stirred at 70° C. for 3 days in a sealed tube. After cooling, the crystals are collected by filtration, dried and mono (6
-amino-6-deoxy)β-cyclodextrin 18
．． 1 g is obtained as a white powder. Yield near 9.8% m, p, 262°C (dec,) I Rv,, xc m-': 3300, 1638, 1
)58.1080.1030(2-b) Bis-(6
-0-mesitylenesulfonyl)-β-cyclodextrin is treated in the same manner as in (2-a) above to obtain bis(6-amino-6-deoxy)-β-cyclodextrin.

Yield near 1% m, p, 245℃ (dec,) I Rv,,, cm m-': 3300, 1630.1)
58.1080.1030 (3-a) Above (2-a
) is suspended in 10- water and 10- tetrahydrofuran, 0.1 g of sodium bicarbonate and 0.21 g of benzenesulfonyl chloride are added, and the mixture is stirred overnight at room temperature. After concentrating the reaction solution, it is crystallized by cooling on ice, and the precipitated crystals are collected by filtration, washed with water and acetone, and dried to obtain mono (6-benzenesulfonylamino-6-deoxy)-
1.0 g of β-cyclodextrin is obtained as a white powder. Yield: 1B, 5% m, p, 233-236°C (de
c, )'II-NMR (DMSO-db) δ: 4.
3-4.6 (m, 61)), 4.7-5.0 (m, 7H
), 5.6-5.9 (m, 14H), 7.4-7.7 (
m, 3H).

7.7-7.9(m, 2H) (3-b) The product of (2-b) and naphthalenesulfonyl chloride were treated in the same manner as in (3-a) above to obtain bis(6-naphthalene). A white powder of sulfonylamino-6-deoxy)-β-cyclodextrin is obtained. Yield near 0% 'tl-NMR (DMSO-d6) δ: 4.4-
5.0 (m), 5.0-6.2 (m).

7.5-8.6 (m) Reference Example 3 (1) Dissolve 4 g of β-cyclodextrin 1) in 1.21 g of pyridine, add 52 g of 2-naphthalenesulfonyl chloride while stirring with water cooling. The reaction solution is heated at room temperature. After stirring for one day, water is added to stop the reaction. The solvent is distilled off from the reaction solution, water 1) is added to the residue, and the mixture is heated. The supernatant was removed by decantation, and the resulting candy-like substance was washed with water and then dissolved by heating in 1.51 g of 70% methanol. The solution to about 1)? a) and filter the precipitate. Furthermore, it was heated and dissolved in 70% methanol 1), the insoluble matter was filtered off, and then left to crystallize. (6-0-
Naphthalenesulfonyl)-β-cyclodextrin 1
:23.0 g of a mixture of 1 was obtained as a white powder. Yield: 13%') l-NMR (DMSO-ds) δ near, 5-8.5 (br m, 24B) (2) Above (1)
The product was treated in the same manner as in Reference Example 2-(2-a), and
Tris(6-amino-6-deoxy)-β-cyclodextrin and tetrakis(6-amino-6-deoxy)-
A 1:1 mixture of β-cyclodextrins is obtained.

Yield: 68.8% m, p, >220°C IRv,, Xcm: 3300.1630.1) 55.
1080.1030(3) The product of (2) above was treated in the same manner as in Reference Example 2-(3-a) to obtain tris(6-benzenesulfonylamino-6-deoxy)-β-cyclodextrin and tetrakis( 1=1 of 6-benzenesulfonylamino-6-deoxy)-β-cyclodextrin
A mixture of is obtained as a white powder.

Yield: 54% '+1-NMR (DMSO-d6) δ: 4.5-5
．． 2(m), 5.3-6.3(br).

7.4-8.0m) Reference Examples 4 to 13 The corresponding starting compounds were treated in the same manner as in Reference Examples 1 to 3 to obtain the compounds listed in Table 2 below.

Experimental example HIVf +I (Principle) Human T-cell leukemia virus type (human T-ce)
Leukemia virus, HTLV-1 (A
TLV)) When MT-4 cells, a persistently infected cell line, are infected with old V, old V rapidly proliferates, and MT-4
It is known that cells die due to cell damage. Therefore, the inhibitory effect of the sample on old V proliferation can be investigated using the number of viable MT-4 cells infected with old V as an index.

(Method) MT-4 cells were incubated with HIV (TALL-1/LAV culture supernatant) at 0.001 TCIDso (median ti
ssue culture 1nfectious d
ose.

50% tissue culture infectious dose)/ce41 at 37°C.
After infecting for 1 hour with
lf serum: Contains 10% fetal bovine serum)
X 10'cell/ml? Float at i degree. Transfer this cell suspension to a 24-well flat bottom culture plate.
A well amount was added and cultured at 37°C in the presence of 15% carbon dioxide for 5 days. After culturing, the number of living cells in the cell suspension was counted by trypan blue staining. IITV of the specimen
The growth inhibitory effect was determined as the concentration of the sample that inhibited 100% of the infectivity and cytopathic effects of HIV on MT4 cells.

(result) The results are shown in Table 3 below.

(Effects of the Invention) As mentioned above, the polysulfate compound of the present invention has an excellent antiretroviral effect, especially an excellent anti-proliferation effect against HIV, and is also characterized by low toxicity and high safety as a medicine. be.

Furthermore, the polysulfate ester compound of the present invention has weak side effects such as anticoagulant effect, which is known from sulfated polysaccharides.

Agent: Patent Attorney Shoji Nakajima 5 children 1'212no

Claims (6)

[Claims] (1) Of the seven D-glucose units that make up β-cyclodextrin, 1 to 7 have a general formula ▲ Numerical formula, chemical formula, table, etc. ▼ (However, R^1 is an alkyl group; a lower alkyl group , a phenyl group optionally substituted with a lower alkoxy group or a halogen atom; a naphthyl group; or a heterocycle containing one or two heteroatoms selected from nitrogen, oxygen, and sulfur and optionally substituted with a lower alkyl group A polysulfate ester of a β-cyclodextrin sulfonamide derivative or a salt thereof, which is substituted with a unit represented by the formula group. (2) R^1 is an alkyl group having 1 to 10 carbon atoms; 1 carbon number
~4 alkyl group, a phenyl group optionally substituted with a C1-4 alkoxy group or a halogen atom; a naphthyl group; or a thienyl group optionally substituted with a C1-4 alkyl group; 2. The compound according to claim 1, which is an isoxazolyl group or a thiazolyl group. (3) The compound according to claim 1 or 2, having 8 to 19 sulfate ester groups. (4) Of the seven D-glucose units that make up β-cyclodextrin, 1 to 7 have a general formula ▲ Numerical formula, chemical formula, table, etc. ▼ (However, R^1 is an alkyl group; a lower alkyl group , a phenyl group optionally substituted with a lower alkoxy group or a halogen atom; a naphthyl group; or a heterocycle containing one or two heteroatoms selected from nitrogen, oxygen, and sulfur and optionally substituted with a lower alkyl group The β-cyclodextrin sulfonamide derivative substituted with a unit represented by the formula group) is reacted with a sulfonating agent, and if desired, the product is converted into a salt thereof. A method for producing a polysulfate ester of a cyclodextrin sulfonamide derivative or a salt thereof. (5) the sulfonating agent is a sulfur trioxide complex;
5. The method according to claim 4, wherein the sulfuric acid is anhydrous sulfuric acid, concentrated sulfuric acid or chlorosulfuric acid. (6) Of the seven D-glucose units that make up β-cyclodextrin, 1 to 7 have a general formula ▲ Numerical formula, chemical formula, table, etc. ▼ (However, R^1 is an alkyl group; a lower alkyl group , a phenyl group optionally substituted with a lower alkoxy group or a halogen atom; a naphthyl group; or a heterocycle containing one or two heteroatoms selected from nitrogen, oxygen, and sulfur and optionally substituted with a lower alkyl group A β-cyclodextrin sulfonamide derivative substituted with a unit represented by the formula group.