RU2794743C1 - Method for manufacturing of imidoamine bis-adduct of maleopimaric acid metil ester with dimethyl-hexaazaperhydropyrene spacer and its use as a drug with cytotoxic activity - Google Patents

Abstract

FIELD: synthesis of biologically active compounds.

SUBSTANCE: method for the production of 2,4,6-trimethyl-5-{2-[2,4,6-trimethyl-5-{2-[2,4,6-trimethyl-1,3,5-dithiazinan-5-il]-ethyl}-1,3,5-thiadiazinan-3-il]-ethyl}-1,3,5-dithiazinan. The invention relates to the field of synthesis of biologically active compounds, specifically to method for the production of bis-adduct of maleopimaric acid methyl ester (MPAME) imidoamine with dimethyl-hexaazaperhydropyrene spacer of formula (1). The essence of the method is the interaction of 2,6-dimethyl-1,4,5,8-tetraazadecaline with N,N,N',N'-tetramethylmethanediamine and MPAME imidoamine in the presence of catalyst NiCl₂ ⋅ 6H₂O at a molecular ratio of 2,6-dimethyl-1,4,5,8-tetraazadecalin : N,N,N',N'- tetramethylmethanediamine : MPAME imidoamine : NiCl₂ ⋅ 6 H₂O = 1 : 4 : 2 : (0.03-0.07) in CH₃OH-H₂O medium at ~20° C and atmospheric pressure for 2.5–3.5 h. The yield of bis-adduct of MPAME imidoamine with dimethyl-hexaazaperhydropyrene spacer (1) is 14-24%.

EFFECT: technical result of the invention is production of bis-adduct of maleopimaric acid methyl ester imidoamine with dimethyl-hexaazaperhydropyrene spacer of formula (1), which has cytotoxic activity against human cancer cells (HTC-116, SH-SY5Y, MCF-7, A549, Jurkat, THP-1).

1 cl, 2 tbl, 5 ex

Description

The present invention relates to the field of synthesis of compounds with biological activity, specifically, to a method for obtaining a bis -adduct of imido-amine of maleopimaric acid methyl ester with a dimethyl-hexaazaperhydropyrene spacer ( 1 ) based on the catalytic condensation reaction of 2,6-dimethyl-1,4,5 ,8-tetraazadecaline with N,N,N',N' -tetramethylmethanediamine and maleopimaric acid methyl ester imidoamine (MEMPC). Compound (1) in vitro exhibits a cytotoxic effect against cell lines of colon adenocarcinoma (HTC-116), human neuroblastoma (SH-SY5Y), breast carcinoma (MCF-7), lung adenocarcinoma (A549), T-cell leukemia ( Jurkat), human monocytic leukemia (THP-1) and can be used as a cytotoxic agent.

A known method of obtaining [MP Bei, AP Yuvchenko, NV Puchkova, Russ. J. Gen. Chem. , 2015, 85, 1034] diimide diacids (2) by the interaction of maleopimaric acid with para- or meta -phenylenediamines at boiling for 8 hours in dichlorobenzene.

The known method does not allow to obtain a bis -adduct of imido-amine methyl ester of maleopimaric acid with dimethyl-hexaazaperhydropyrene spacer (1).

A known method of obtaining [Rakhimova, EB; Kirsanov, V.Yu.; Tret'yakova, EV; Khalilov, L.M.; Ibragimov, A.G.; Dzhemileva, L.U.; D'yakonov, V. A.; Dzhemilev, UM RSC Advances 2020, 10, 36, 21039] of dimeric hexaazaperhydrodibenzotetracenes (3) by cyclocondensation of amine derivatives of MEMPC with formaldehyde and tetraazaperhydrotetracene in the presence of Ymmm zeolite in the H-form as a catalyst.

The known method does not allow to obtain a bis -adduct of imido-amine methyl ester of maleopimaric acid with dimethyl-hexaazaperhydropyrene spacer (1).

A known method of obtaining [Rakhimova, EB; Kirsanov, V.Yu.; Meshcheryakova, E.S.; Khalilov, L.M.; Kutepov, B.I.; Ibragimov, AG; Dzhemilev, UM Tetrahedron 2017, 73 , 6880] dialkyl-substituted (4) and dicycloalkyl-substituted (5) 4,9-dimethyl-2,3а,5а,7,8а,10а-hexaazaperhydropyrenes by intermolecular heterocyclization N,N - bis (methoxymethyl) - N -alkylamines or recyclization of 1,3,5-tricycloalkyl-1,3,5-triazinanes with 2,6-dimethyl-1,4,5,8-tetraazadecalin under the action of catalysts based on salts of d- and f- elements.

The known method does not allow to obtain a bis -adduct of imido-amine methyl ester of maleopimaric acid with dimethyl-hexaazaperhydropyrene spacer (1).

A known method of obtaining [Rakhimova, EB; Kirsanov, V.Yu.; Ibragimov, AG; Dzhemilev, UM Russ. J. Org. Chem. 2018, 54 , 1085] diaryl-substituted 4,9-dimethyl-2,3а,5а,7,8а,10а-hexaazaperhydropyrenes (6) by cyclocondensation of arylamines with formaldehyde and 2,6-dimethyl-1,4,5,8-tetraazadecalin in the presence of a catalyst based on Yb (III).

The known method does not allow to obtain a bis -adduct of imido-amine methyl ester of maleopimaric acid with dimethyl-hexaazaperhydropyrene spacer (1).

Known are [Elena V. Tretyakova, Irina E. Smirnova, Oxana B. Kazakova, Genrikh A. Tolstikov, Nadejda P. Yavorskaya, Irina S. Golubeva, Rujena B. Pugacheva, Galina N. Apryshko, Vladimir V. Poroikov. Bioorganic & Medicinal Chemistry 2014, 22, 6481] in vitro antitumor activity of compound (7).

Known [Elena V. Tretyakova, Irina E. Smirnova, Elena V. Salimova, Victor N. Odinokov. Bioorganic & Medicinal Chemistry 2015, 23, 6543] high activity of compound (8) against human papillomavirus (HPV-11 strain).

Known for [E.V. Tretyakova, E.V. Salimova, L.V. Parfenov. Bioorganic Chemistry , 2018, 44, 5, 554] moderate in vitro antitumor activity of the cyanethyl derivative of maleopimaric acid methyl ester (9) against prostate cancer cells (PC-3) .

Syntheses of these compounds are either multistage or based on the use of expensive starting reagents.

Thus, there is no information in the literature about the method of obtaining the imidoamine bis -adduct of maleopimaric acid methyl ester with dimethyl-hexaazaperhydropyrene spacer (1) and its biological properties.

A new method is proposed for the preparation of a bis -adduct of imido-amine of maleopimaric acid methyl ester with dimethyl-hexaazaperhydropyrene spacer (1) in one preparative stage from available starting reagents. The compound of formula (1) can be used as an agent with cytotoxic activity.

The essence of the method lies in the interaction of 2,6-dimethyl-1,4,5,8-tetraazadecalin with N,N,N',N'- tetramethylmethanediamine and imidoamine MEMPK in the presence of NiCl ₂ ⋅6H ₂ O catalyst, taken in molar ratio 2,6-dimethyl-1,4,5,8-tetraazadecalin : N,N,N',N' -tetramethylmethanediamine : imidoamine MEMPK : NiCl ₂ ⋅6H ₂ O = 1 : 4 : 2 : (0.03- 0.07), preferably 1 : 4 : 2 : 0.05. The reaction mixture was stirred for 2.5–3.5 h at a temperature of ~20°C and atmospheric pressure in CH ₃ OH–H ₂ O. The yield of the bis -adduct of imidoamine MEMPC with dimethyl-hexaazaperhydropyrene spacer (1) was 14–24%. The reaction proceeds according to the scheme:

Imido-amine MEMPC receive according to the method described in [H. Wang, H. Wang, G. Zhou, Polym. Int. , 2011, 60, 557]. 2,6-Dimethyl-1,4,5,8-tetraazadecalin is obtained by the method described in [R. Mullera, W. Philipsborna, L. Schleiferh, P. Aped, B. Fuchs, Tetrahedron 1991, 47, 1013]. Bis -adduct of imidoamine MEMPK with dimethyl-hexaazaperhydropyrene spacer ( 1 ) is formed only with the participation of 2,6-dimethyl-1,4,5,8-tetraazadecaline, tetramethylmethanediamine and imidoamine MEMPK, taken in a molar ratio of 1 : 4 : 2 (stoichiometric quantities). At a different ratio of the initial reagents, the yield of the target product decreases ( 1 ). The reaction does not proceed without a catalyst. Carrying out this reaction in the presence of a NiCl ₂ ⋅6H ₂ O catalyst is more than 7 mol. % does not lead to a significant increase in the yield of the target product (1). The use of catalyst NiCl ₂ ⋅6H ₂ O less than 3 mol. % reduces the yield (1), which is possibly associated with a decrease in catalytically active centers in the reaction mass. The reactions were carried out at a temperature of 20 ^about C. At temperatures above 20°C (for example, 60°C) energy costs increase, and at temperatures below 20°C (for example, at 0°C) the reaction rate decreases. The experiments were carried out in CH ₃ OH-H ₂ O medium, because the starting compounds dissolve well in them.

The cytotoxicity of compound (1) was evaluated on cancer cell lines (HTC-116, SH-SY5Y, MCF-7, A549, Jurkat, THP-1, HepG2) and conditionally normal human cells (HEK293) using the MTT test (colorimetric test for assessing metabolic cell activity). Cells were incubated in the presence of compound (1) for 48 hours. 5-fluorouracil served as a control. As can be seen from Table 1, compound ( 1 ) inhibits the viability of cancer cells at IC ₅₀ values from 37 to 73 μM, depending on the tested cell line, and exhibits a moderate cytotoxic effect.

Table 1. Effect of bis -adduct of imidoamine MEMPC with dimethyl-hexaazaperhydropyrene spacer ( 1 ) on the viability of cancer cells in vitro . HEK293
IC ₅₀ /µM HTC-116
IC ₅₀ /µM SH-SY5Y
IC ₅₀ /µM MCF-7
IC ₅₀ /µM 1 53.5± 0.4 68.4±0.5 55.8±0.5 39.5±0.2 5-fluorouracil 6.32 ± 0.71 2.38±0.9 1.16±0.3 1.0±0.04 A-549
IC ₅₀ /µM Jurkat
IC ₅₀ /µM THP-1
IC ₅₀ /µM HepG2
IC ₅₀ /µM 1 73.3±0.8 37.1±0.2 46.8±0.5 > 100 5-fluorouracil 0.28±0.02 0.67±0.10 4.3±0.8 3.86±0.7

Significant differences of the proposed method:

In a known method, the reaction proceeds with the participation as initial reagents 1,6,7,12-tetraazaperhydrotetracene, formaldehyde and amino derivatives MEMPC in the presence of Ymmm zeolite in the H-form as a catalyst.

In the proposed method, the reaction proceeds with the participation of 2,6-dimethyl-1,4,5,8-tetraazadecalin, tetramethylmethanediamine and imidoamine MEMPK as initial reagents in the presence of a NiCl ₂ ⋅ 6H ₂ O catalyst. Unlike the known one, the proposed method allows to obtain a bis -adduct of imido-amine MEMPC with dimethyl-hexaazaperhydropyrene spacer (1) with cytotoxic activity.

The method is illustrated with examples:

EXAMPLE 1. 0.17 g (1 mmol) of 2,6-dimethyl-1,4,5,8-tetraazadecalin in 5 ml of H ₂ O, 0.41 g (4.00 mmol) tetramethylmethanediamine and 0.012 g (0.05 mmol) catalyst NiCl ₂ ⋅6H ₂ O, then add 0.43 g (2.00 mmol) imidoamine MEMPC in 10 ml MeOH. The reaction mixture was stirred for 3 hours at room temperature. The resulting precipitate is filtered off, washed twice with methanol (2x5 ml) and an individual bis -adduct of imido-amine MEMPC with a dimethyl-hexaazaperhydropyrene spacer, namely dimethyl- N',N'' -[4,9-dimethyloctahydro-1 H ,6 H -2,3a,5a,7,8a,10a-hexaazapyrene-2,7-diylbis(ethane- N',N'' -diyl)]bis(13'-isopropyl-4',10'-dimethyl-23',24'-dioxohexadecahydro-8',12'-ethenonaphtho[2,1- e ]isoindole-4'-carboxylate) in 21% yield.

Examples 2-5, confirming the method, performed similarly to pr. 1 and are given in table. 2.

table 2 №№
p/p The ratio of 2,6-dimethyl-1,4,5,8-tetraazadecalin : tetramethylmethanediamine :
imido-amine MEMPK : NiCl₂⋅6H₂O, mmol Reaction time,
hour Yield ( 1 ), % 1 1:4:2:0.05 3 21 2 1:4:2:0.03 3 14 3 1:4:2:0.07 3 24 4 1:4:2:0.05 2.5 17 5 1:4:2:0.05 3.5 23

All experiments were carried out in CH ₃ OH-H ₂ O

at room temperature (~20°C).

Spectral characteristics of bis -adduct of imido-amine MEMPC with dimethyl-hexaazaperhydropyrene spacer:

^1H NMR spectrum, δ, ppm ( J , Hz): 0.58 s (6H, CH ₃ , H-20',20''), 0.89-0.98 m (2H, CH ₂ , H _ax -1',1''; 12H, CH ₃ , H -16',16'',17',17''; 6H, CH ₃ , H-11,12), 1.15 s (6H, CH ₃ , H-19',19''), 1.16-1.79 m ( 20Н, CH ₂ , H _eq -1',1'', H _ax,eq -2',2'',3',3'',6',6'',11',11'', H _ax -7',7''; 4H, CH, H-5',5'',9',9''), 1.82-1.97 m (2H, CH ₂ , H _a -5.10), 2.10-2.19 m (2H, CH, H-15',15''), 2.30-2.42 m (6H, CH ₂ , H _b -5.10, H-26',26''; 6H, CH, H-22' ,22′′, H-4.9, H-10b,10c), 2.50 d (2H, CH ₂ , H _eq -7′,7′′, ^2J = 13), 2.73-2.78 m (2H, CH , H-21′ _, 21′′), 2.88-3.05 _m (4H, CH ₂ , H-27',27''), 3.51-3.58 m (2H, CH ₂ , H _b -1.6), 3.67 s (6H, OCH ₃ , H-25',25'' ), 3.92-3.98 m (2H, CH ₂ , H _b -3.8), 5.37 s (2H, CH, H-14',14'').

^13С NMR spectrum, δ, ppm: 15.63 (С-20',20''), 16.75 (С-11.12, С-19',19''), 17.04 (С-2',2 ′′), 20.09 (С-16′,16′), 20.71 (С-17′,17′′), 21.78 (С-6′,6′′), 27.60 (С-11′,11′′) , 32.65 (С-15′,15′′), 35.32 (С-7′,7′′), 35.57 (С-12′,12′′), 36.56 (С-27′,27′′), 36.69 (С-3′,3′′), 37.67 (С-10′,10′′), 38.16 (С-1′,1′′), 40.65 (С-8′,8′′), 45.01 (С -21′,21′′), 47.16 (С-4′,4′′), 49.53 (С-5′,5′′), 49.61 (С-26′,26′′), 51.06 (С-4 ,9), 51.95 (С-22′,22′′), 52.36 (С-25′,25′′), 54.15 (С-9′,9′′), 55.98 (С-5.10), 70.16 (C-3.8), 73.50 (C-1.6), 82.63 (C-10b,10c), 124.38 (C-14',14''), 146.95 (C-13',13''), 177.55 (C-24',24''), 178.65 (C-23',23''), 179.24 (C-18',18'').

MALDI TOF/TOF: m/z (%) = 1129.718 [ M -H] ⁺ (100) and 1169.917 [ M +K] ⁺ (100).

NMR spectra ( ¹ H, ¹³ C) were recorded on a Bruker Avance 500 spectrometer (125.78 MHz for ¹³ C nuclei and 500.17 MHz for ¹ H nuclei) according to standard Bruker procedures, internal standard Me ₄ Si, solvent CDCl ₃ . Mass spectra were obtained on a Bruker MALDI TOF/TOF AUTOFLEX III instrument.

Claims (3)

Method for the preparation of imidoamine bis -adduct of maleopimaric acid methyl ester (MEMPC) with dimethyl-hexaazaperhydropyrene spacer of formula (1)

, characterized in that 2,6-dimethyl-1,4,5,8-tetraazadecalin is reacted with N,N,N',N' -tetramethylmethanediamine and imidoamine MEMPC in CH ₃ OH-H ₂ O in the presence of a catalyst NiCl ₂ 6H ₂ O at a molar ratio of 2,6-dimethyl-1,4,5,8-tetraazadecalin: N,N,N',N'- tetramethylmethanediamine:imido-amine MEMPC:NiCl ₂ 6H ₂ O=1 :4:2:(0.03-0.07) at room (~20°C) temperature and atmospheric pressure for 2.5 - 3.5 hours.