RU2357960C1 - Immunotropic n, n '-(sulphonyldi-1,4-phenylene)bis[(n",n'"-dimethyl)methyliminomethane]1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5-pyrimidinesulphonate - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention refers to new biologically active high-immunotropic compound -N,N'-(sulphonyldi-1,4-phenylene)bis[(N",N'"-dimethyl)methyliminomethane]1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5-pyrimidinesulphonate of formula stated below to be used in treatment of, e.g., patients suffering from leprosy, allergic dermatosis, dermatitis herpetiformis.

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EFFECT: new compound is characterised with useful biological activity.

2 tbl, 1 ex

Description

The invention relates to a new biologically active compound, which belongs to the group of drugs used for the treatment of patients with leprosy, allergic dermatoses, herpetiform dermatitis (diutsifon, dimotsifon).

Known analogs of the claimed compounds diuciphon (SU 459228) and dimocipon (RU 2200550) have anti-leprosy activity (A. S. USSR N 459228), but do not show anti-tuberculosis activity. In addition, the listed analogues are not sufficiently bioavailable and require the use of large doses to achieve a therapeutic effect.

Also known is the compound proposed by the authors of patent RU 2136668 and international application PCT / RU98 / 00314 (publication number WO 99/18083), N, N '- (sulfonyldi-1,4-phenylene) bis (N ", N" -dimethylformamidine) 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5-pyrimidine sulfonate, corresponding to the formula

In accordance with the description of this invention, this drug has advantages over ducifon: it is less toxic, active against leprosy mycobacteria, tuberculosis and opportunistic atypical mycobacteria, has a high immunotropic effect on microorganisms (stimulation index is 3 times higher than that of duciphone) . In addition, he takes part in the metabolism of the cell, stimulates its function 1.5-2 times better than diutsifon.

A significant disadvantage of the compound proposed by the invention according to the patent of the Russian Federation 2136668 is the dependence of the immunotropic activity and a number of other biomedical parameters of the drug samples on difficult to control, and mainly not detected parameters during synthesis by the method described in the description of patent RU No. 2136668 (loading speed of components the reaction mixture, the intensity of mixing, slight temperature fluctuations within the limits allowed by the method according to the patent).

The technical result of the invention is to obtain compounds with higher immunotropic activity.

The technical result is achieved by the compound N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4- dioxo-5-pyrimidine sulfonate,

possessing immunotropic, antimycobacterial activity and stimulating cellular metabolism.

According to the claimed invention, N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4 -dioxo-5-pyrimidine sulfonate is obtained with a high degree of purity due to the use of an organic base as a proton acceptor in the synthesis process.

N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methylaminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5- pyrimidine sulfonate was tested for immunotropic activity in the laboratory of drug toxicology, Research Institute of Pharmacology RAMS.

Study of the immunotropic activity of N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo -5-pyrimidine sulfonate was carried out according to the "Guidelines for assessing the immunotoxic effects of pharmacological agents" (1) and the "Guidelines for assessing the immunotropic activity of pharmacological substances" M., 2005

N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5 preparation -pyrimidine sulfonate, is a derivative of diphenyl sulfone. It is known that the effect of diphenylsulfone is directed mainly to the cellular immune response. N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5 preparation -pyrimidine sulfonate selectively inhibits the synthesis of IgE, while the level of IgG, A and M did not decrease, and in cases where IgG, A and M were initially low, their rise was noted, i.e. the drug showed a truly immunomodulating effect. Under the influence of N, N '- (sulfonyldi-1,4-phenylene) bis [(N ”, N'" - dimethyl) methylaminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo 5-pyrimidine sulfonate, activation of IL-2 producing cells occurs and, as a result, a significant increase in their production of this cytokine leads to stimulation of type I T-helpers (Th1 response) .An increase in the number of CD25 lymphocytes having an IL-2 receptor is observed. as well as an increase in the number of natural killers and an increase in antibody-dependent cellular cytotoxicity. In this regard, the effect of the sample N, N '- (sulfonyldi-1,4-phenyl m) bis [(N ", N '" - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5-pyrimidinesulfonate to the cellular immune response.

Dose N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methylaminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5 -pyrimidine sulfonate for humans is 0.2 g. After recalculation, taking into account species sensitivity, weight and body surface according to Freireich EJ, the dose for mice was 26 mg / kg.

As a control, a sample of N, N '- (sulfonyldi-1,4-phenylene) bis (N ”, N" -dimethylformamidine) 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5 was used -pyrimidine sulfonate.

The influence of the control and test samples of the drug on the cellular immune response was studied by the delayed-type hypersensitivity reaction in experiments on mice F ₁ hybrids (CBAxC57BL / 6). The control sample N, N '- (sulfonyldi-1,4-phenylene) bis (N ", N" -dimethylformamidine) 1,2,3,4-tetrahydro-6- was orally administered to the animals of the first experimental group for 3 days 26 mg / kg methyl 2,4-dioxo-5-pyrimidine sulfonate; mice of the second group were similarly injected with the test sample N, N '- (sulfonyldi-1,4-phenylene) bis [(N ”, N”' dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5-pyrimidine sulfonate at a dose of 26 mg / kg.

The control animals were injected with an appropriate volume of a 1% starch solution. Before the last administration of the drug, mice of the control and experimental groups were immunized subcutaneously with 1 × 10 ⁷ EB in a volume of 100 μl. A resolving dose of antigen, 1 × 10 EB in a volume of 20 μl, was administered on day 5 after sensitization, a sub - aponeurotic - plate of the right hind limb, and 20 μl of physiological saline in the contralateral paw. The intensity of the inflammatory reaction was taken into account 24 hours after the resolving dose of the antigen. Mice were scored, both legs were cut off at the level of the ankle joint and weighed on a torsion balance. Then calculated the reaction index (And _p ) by the formula:

where R _op - the mass of the foot of the hind paw, in the pad of which was introduced EB;

P _to - the mass of the foot of the control paw.

Table 1 shows the reaction indices obtained as a result of experiments.

Table 1. Test results (reaction indices) of N, N '- (sulfonyldi-1,4-phenylene) bis (N ”, N” -dimethylformamidine) 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo samples -5-pyrimidine sulfonate (control sample) and N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6- methyl 2,4-dioxo-5-pyrimidine sulfonate (test sample) Animal number The control Control sample 26 mg / kg Test sample 26 mg / kg one 16.1 41.9 65,2 2 14.6 27.8 42.0 3 19.6 47.8 29.2 four 31.7 28,2 43.1 5 20.7 42,2 43,4 6 25,2 37.1 63,4 7 21.9 41.7 31.3 8 20,0 35.3 35,2 9 19.2 35.0 42.5 10 24.9 39.2 46.0

The obtained values were processed by the method of variation statistics using t-student criterion. The results of statistical processing are presented in table 2.

Table 2. The effect of the drug on the cellular immune response (HRT) Dose of the drug

Reaction Index:

Significance level The control 21.4 ± 1.6
n = 10 Control sample 26 mg / kg 37.6 ± 2.0
n = 10 P <0.01 Test sample 26 mg / kg 44.1 ± 3.8
n = 10 P <0.01 Note: n is the number of animals in the group.

The results presented in table 2 indicate that triple oral administration of the test samples at a dose of 26 mg / kg led to a pronounced stimulation of cellular immunity. A control sample of the drug stimulated cellular immunity by 75.7% (1.8 times) compared with the control group. The test sample stimulated a T-cell immune response by 106.1% (2.1 times) compared with the control. The obtained results indicate the presence of immunotropic activity in the studied samples of N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methyliminomethane] -1,2,3,4-tetrahydro-6 methyl 2,4-dioxo-5-pyrimidine sulfonate.

Thus, the results obtained in the study of two samples of the drug indicate an increase in the activity of the test sample of the drug compared with the control sample.

EXAMPLE

Preparation of N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N'" - dimethyl) methylaminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo-5 -pyrimidinesulfonate

To a solution of 4.96 g (0.02 mol) of 4,4'-diaminodiphenyl sulfone in 20 ml of dimethylformamide was added 4.05 g (0.04 mol) of triethylamine. Then, with stirring in small portions, 9.4 g (0.042 mol) of 6-methyluracil sulfochloride are added, keeping the temperature not higher than 30-35 ° С. After that, the reaction mixture was stirred for 2 hours at 50 ° C. The cooled reaction mass is poured into 100 ml of chloroform, stirred for 1.5 hours. The precipitate formed is filtered off. 60 ml of ethyl alcohol are added to the resulting product, heated for one hour, filtered, dried at 40 ° C in vacuo. Received 9.6 g (80%) of a yellowish substance.

PMR spectrum (300 MHz, d ⁶ DMSO):

Filmed at 20 ° C 2.00 (CH ₃ 6-methyluracil), 2.38 s (3 H, CH ₃ ); 3.27 s (6 H, NCH ₃ ); 3.38 s (6 H, NCH ₃ ); 5.32 (H 6-methyluracil); 7.72 d (4H, aromatic); 8.02d (4H, aromatic); 8.89 s (2H, N = CH ), 10.63 s and 10.87 s ( NH ); 11.33 N ⁺ H.

Filmed at 80 ° C: 2.00 (CH ₃ 6-methyluracil), 2.42 s (3 H, CH ₃ ); 3.29 s (6 H, NCH ₃ ); 3.36 s (6 H, NCH ₃ ); 5.32 (H 6-methyluracil); 7.65 d (4H, aromatic); 7.97d (4H, aromatic); 8.72 s (2H, N = CH ), 10.67 s and 10.87 s ( NH ); 11.73 N ⁺ H.

In the PMR spectrum, taken upon heating (80 ° C), the signals of methyl protons at nitrogen atoms do not change their position. This confirms that the methyl groups are located at different nitrogen atoms in the formamidine component of the claimed complex.

In the NMR spectrum of the control sample (N, N '- (sulfonyldi-1,4-phenylene) bis (N ", N" -dimethylformamidine) 1,2,3,4-tetrahydro-6-methyl-2,4-dioxo 5-pyrimidine sulfonate), taken by heating, the signals of methyl protons merge (3.32 ppm).

Control sample:

PMR spectrum (300 MHz, d ⁶ DMSO):

Taken at 20 ° C: 2.38 s (3 N, CH ₃ ); 3.25 s (6 H, NCH ₃ ); 3.35 s (6 H, NCH ₃ ); 7.65 d (4H, aromatic); 8.01d (4H, aromatic); 8.78 s (2H, N = CH ), 10.68 and 10.90 ( NH ).

Taken at 80 ° C: 2.42 s (3 N, CH ₃ ); 3.32 s (12 H, NCH ₃ ); 7.65 d (4H, aromatic); 7.97 d (4H, aromatic); 8.68 s (2H, N- CH ).

The structure of the claimed compounds is confirmed by UV, IR and NMR spectroscopy, mass spectrometry.

¹ H NMR spectra were recorded on Bruker spectrometers (300 MHz) in deuterated dimethyl sulfoxide at 20 ° C and 80 ° C.

Mass spectrometric analysis was performed on an MX-5303 instrument equipped with an electrospray ionization source with orthogonal ion input and a time-of-flight mass analyzer (ESI-o-TOF).

IR spectra were recorded on a Bruker instrument (FT-IR spectrometer).

UV spectra were recorded on an SF-46 spectrophotometer.

In the PMR spectrum of the claimed complex N, N '- (sulfonyldi-1,4-phenylene) bis [(N ", N"' - dimethyl) methyliminomethane] 1,2,3,4-tetrahydro-6-methyl-2,4 -dioxo-5-pyrimidine sulfonate, taken by heating (80 ° C) the signals of methyl protons at nitrogen atoms do not change their position. This confirms that the methyl groups are located at different nitrogen atoms in the formamidine component of the claimed complex.

PMR spectrum (300 MHz, d ⁶ DMSO):

at 20 ° C: 2.38 s (3 N, CH ₃ ); 3.27 s (6 H, NCH ₃ ); 3.38 s (6 H, NCH ₃ ); 7.72 d (4H, aromatic); 8.02d (4H, aromatic); 8.89 s (2H, N = CH ), 10.63 s and 10.87 s ( NH ); 11.33 N ⁺ H.

at 80 ° C: 2.42 s (3 N, CH ₃ ); 3.29 s (6 H, NCH ₃ ); 3.36 s (6 H, NCH ₃ ); 7.65 d (4H, aromatic); 7.97d (4H, aromatic); 8.72 s (2H, N = CH ).

In the NMR spectrum of the control sample (N, N '- (sulfonyldi-1,4-phenylene) bis (N ”, N” -dimethylformamidine) 1,2,3,4-tetrahydro-6-metal-2,4-dioxo 5-pyrimidine sulfonate) (STANDART), taken when heated, the signals of metal protons merge (3.32 ppm).

PMR spectrum (300 MHz, d ⁶ DMSO):

Taken at 20 ° C: 2.38 s (3 N, CH ₃ ); 3.25 s (6 H, NCH ₃ ); 3.35 s (6 H, NCH ₃ ); 7.65 d (4H, aromatic); 8.01d (4H, aromatic); 8.78 s (2H, N = CH ), 10.68 and 10.90 ( NH ).

Taken at 80 ° C: 2.42 s (3 N, CH ₃ ); 3.32 s (12 H, NCH ₃ ); 7.65 d (4H, aromatic); 7.97 d (4H, aromatic); 8.68 s (2H, N = CH ).

The composition of the complex was confirmed by HPLC.

Claims (2)

1. N, N '- (Sulfonyldi-1,4-phenylene) bis [(N'',N''' - dimethyl) methyliminomethane] 1,2,3,4-
tetrahydro-6-methyl-2,4-dioxo-5-pyrimidine sulfonate of the formula

2. The compound according to claim 1, having immunotropic activity.