RU2794745C1 - Method for manufacturing of terpenoid dimer with perhydrotetracene spacer and its use as with a 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 organic chemistry, sprcifically to the method of terpenoid dimer with perhydrotetracene spacer (1), which includes interaction of cis- 1,6,7,12-tetraazaperhydrotetracene with N,N,N',N' -tetramethylmethanediamine and imido- amine MEMPA in CH₃OH medium in the presence of catalyst NiCl₂⋅ 6H₂O at the molar ratio of cis- 1,6,7,12-tetraazaperhydrotetracene: N,N,N',N' -tetramethylmethanediamine : imidoamine MEMPA: NiCl₂⋅ 6H₂O = 1: 4: 2: (0.03-0.07) at room temperature (~20° C) temperature and atmospheric pressure for 2.5-3.5 hours.

EFFECT: technical result: a method has been developed for manufacturing of terpenoid dimer with perhydrotetracene spacer (1), which has cytotoxic activity against human cancer cells (HTC-116, SH-SY5Y, MCF-7, Jurkat, THP-1).

1 cl, 2 tbl, 1 ex

Description

The present invention relates to the field of synthesis of compounds with biological activity, specifically, to a method for obtaining a terpenoid dimer with a perhydrotetracene spacer ( 1 ) based on the catalytic condensation reaction of cis - 1,6,7,12-tetraazaperhydrotetracene 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), T-cell leukemia (Jurkat), human monocytic leukemia (THP-1) and can be used as a cytotoxic agent.

A known method of obtaining [RMB Carrilho, MM Pereira, MJSM Moreno, A. Takács, L. Kollár, Tetrahedron Lett. , 2013, 54, 2763] of steroidal dicarboxamides (2) by reacting 17-iodo-5α-androst-16-ene with aliphatic and aromatic diamines in an autoclave in the presence of a Pd(OAc) ₂ /PPh ₃ palladium-phosphine catalyst.

The known method does not allow to obtain a terpenoid dimer with perhydrotetracene spacer (1).

A known method of obtaining [MP Bei, AP Yuvchenko, NV Puchkova, Russ. J. Gen. Chem. , 2015, 85, 1034] diimide diacids (3) 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 terpenoid dimer with perhydrotetracene 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] dimeric perhydrohexaazadibenzotetracenes (4) by cyclocondensation of amine derivatives of MEMPC with formaldehyde and trans -tetraazaperhydrotetracene in the presence of Ymmm zeolite in the H-form as a catalyst.

The known method does not allow to obtain a terpenoid dimer with perhydrotetracene spacer (1).

Known [EV Tretyakova, IE Smirnova, EV Salimova, TM Pashkova, OL Kartashova, VN Odinokov, LV Parfenova, Russ. J. Bioorg. Chem. , 2017, 43, 317] moderate antibacterial activity of the maleopimaric acid derivative (5) against Staphylococcus aureus.

Known are [OB Kazakova, EV Tret'yakova, OS Kukovinets, GA Tolstikov, TI Nazyrov, IV Chudov, AF Ismagilova, Russ. J. Bioorg. Chem. , 2010, 36, 762] anti-inflammatory and antiulcer properties of maleopimaric acid derivative (6).

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 (7) against prostate cancer cells (PC-3) .

Known for [G. Yao, M. Ye, R. Huang, Y. Li, Y. Zhu, Y. Pan, Z. Liao, H. Wang, Bioorg. Med. Chem. Lett. , 2013, 23 , 6755] antiproliferative activity of maleopimaric acid methyl ester N -arylimide ( 8 ) against NCI, A549, HepG-2, MGC-803, Hct-116 cell lines.

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 a terpenoid dimer with a perhydrotetracene spacer (1) and its biological properties.

A new method is proposed for the preparation of a terpenoid dimer with a perhydrotetracene spacer (1) in one preparative step from available starting reagents. The compound of formula (1) can be used as an agent with cytotoxic activity.

The essence of the method consists in the interaction of cis- 1,6,7,12-tetraazaperhydrotetracene with N,N,N',N' -tetramethylmethanediamine and imidoamine MEMPC in the presence of a catalyst NiCl ₂ ⋅6H ₂ O, taken in the molar ratio of cis- 1 ,6,7,12-tetraazaperhydrotetracene : 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 a CH ₃ OH medium. The yield of terpenoid dimer with perhydrotetracene spacer (1) is 15-22%. 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]. cis- 1,6,7,12-Tetraazaperhydrotetracene is obtained by the method described in [EB Rakhimova, V. Yu. Kirsanov, ES Mescheryakova, LM Khalilov, AG Ibragimov, UM Dzhemilev, Synlett, 2018, 29, 1861]. A terpenoid dimer with a perhydrotetracene spacer ( 1 ) is formed only with the participation of cis- 1,6,7,12-tetraazaperhydrotetracene, tetramethylmethanediamine, and imidoamine MEMPC, taken in a molar ratio of 1 : 4 : 2 (stoichiometric amounts). 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°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 medium, because the starting compounds dissolve well in it.

The cytotoxicity of compound ( 1 ) was evaluated on cancer cell lines (HepG2, HTC-116, SH-SY5Y, MCF-7, A-549, Jurkat, THP-1) and conditionally normal human cells (HEK293) by the MTT assay (colorimetric test for assessment of metabolic activity of cells). Cells were incubated in the presence of compound ( 1 ) for 48 h. 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 45 to 82 μM, depending on the tested cell line, and exhibits a moderate cytotoxic effect.

Table 1. Effect of terpenoid dimer with perhydrotetracene spacer ( 1 ) on cancer cell viability in vitro . HEK293
IC ₅₀ /µM HepG2
IC ₅₀ /µM HTC-116
IC ₅₀ /µM SH-SY5Y
IC ₅₀ /µM 1 68.8±1.6 > 100 82.3±0.1 77.9±0.2 5-fluorouracil 6.32 ± 0.71 3.86±0.7 2.38±0.9 1.16±0.3 MCF-7
IC ₅₀ /µM A-549
IC ₅₀ /µM Jurkat
IC ₅₀ /µM THP-1
IC ₅₀ /µM 1 58.6±1.4 > 100 45.9±0.3 60.0±2.9 5-fluorouracil 1.0±0.04 0.28±0.02 0.67±0.10 4.3±0.8

Significant differences of the proposed method:

In a known method, the reaction proceeds with the participation as initial reagents trance-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 cis- 1,6,7,12-tetraazaperhydrotetracene, tetramethylmethanediamine and imidoamine MEMPK in the presence of a NiCl ₂ ⋅ 6H ₂ O catalyst as initial reagents. In contrast to the known one, the proposed method makes it possible to obtain a terpenoid dimer with perhydrotetracene spacer (1).

The method is illustrated with examples:

EXAMPLE 1. 0.43 g (2.00 mmol) of imidoamine MEMPC in 10 ml of MeOH, 0.13 g (1 mmol) of cis- 1,6,7,12 -tetraazaperhydrotetracene, 0.41 g (4.00 mmol) tetramethylmethanediamine, and 0.012 g (0.05 mmol) NiCl ₂ ⋅6H ₂ O catalyst. The reaction mixture was stirred for 3 h at room temperature. The resulting precipitate is filtered off, washed twice with methanol (2x5 ml) and an individual terpenoid dimer with a perhydrotetracene spacer is obtained, namely dimethyl N',N'' -[(3b S* ,7a R* ,10b R* ,14a S* )-tetradecahydro -1 H ,8 H -2,3a,7b,9,10a,14b-hexaazadibenzo[ fg,op ]tetracene-2,9-diylbis(ethane -N',N'' -diyl)]bis(13'- isopropyl-4',10'-dimethyl-23',24'-dioxohexadecahydro-8',12'-ethenonaphtho[2,1- e ]isoindole-4'-carboxylate) in 19% yield.

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

table 2 №№
p/n The ratio of cis-1,6,7,12-tetraaza-perhydrotetracene: tetramethylmethanediamine:
imido-amine MEMPK : NiCl ₂ ⋅6H ₂ O, mmol Reaction time,
hour Yield (1), % 1 1:4:2:0.05 3 19 2 1:4:2:0.03 3 15 3 1:4:2:0.07 3 22 4 1:4:2:0.05 2.5 17 5 1:4:2:0.05 3.5 21

All experiments were carried out in a CH ₃ OH medium at room temperature (~20°C).

Spectral characteristics of terpenoid dimer with perhydrotetracene spacer:

^1H NMR spectrum, δ, ppm: 0.60 s (6H, CH ₃ , H-20',20''), 0.92-1.02 m (2H, CH ₂ , H _ax -1',1''; 12H, CH ₃ , H-16',16'',17',17''), 1.16 s (6H, CH ₃ , H-19',19''), 1.21-1.94 m (36H, CH ₂ , H _eq -1′,1′′, H _ax,eq -2′,2′′,3′,3′′,6′,6′′, H _ax -7′,7′′, H _ax,eq -11',11''; H _a,b -4,5,6,7,11,12,13,14; 4H, CH, H-5',5'',9',9''), 2.16-2.20 m (2H, CH, H-15',15''), 2.42-2.53 m (2H, CH ₂ , H _eq -7',7'', 6H, CH, H-22',22' ′, H-3b,7a,10b,14a), 2.79 br. s (2H, CH, H-21',21'', 2H, CH ₂ , H _a -1.8), 2.82-2.98 m (4H, CH ₂ , H-26',26''; 2H, CH , H-14c,14d), 3.04 s (2H, CH, H-12',12''), 3.41-3.60 m (6H, CH ₂ , H _a -3.10, H-27',27'' ), 3.68 s (6H, OCH ₃ , H-25',25''), 3.83-3.88 m (4H, CH ₂ , H _b -1,3,8,10), 5.38 s (2H, CH, H -14′,14′′).

^13С NMR spectrum, δ, ppm: 15.65 (С-20′,20′′), 16.77 (С-19′,19′′), 17.06 (С-2′,2′′), 19.96 ( C-5,7,12,14), 20.06 (С-16′,16′′), 20.71 (С-17′,17′′), 21.79 (С-6′,6′′), 25.49 (С -6.13), 27.59 (С-11′,11′′), 27.83 (С-4.11), 32.67 (С-15′,15′′), 35.35 (С-7′,7′′) , 35.67 (С-12′,12′′), 36.71 (С-3′,3′′, С-27′,27′′), 37.69 (С-10′,10′′), 38.17 (С- 1′,1′′), 40.69 (С-8′,8′′), 44.99 (С-21′,21′′), 47.17 (С-4′,4′′), 49.55 (С-5′ ,5′′), 50.02 (С-26′,26′′), 51.95 (С-22′,22′′), 52.38 (С-25′,25′′), 54.21 (С-9′,9 ′′), 57.47 (C-3b,10b), 59.20 (C-7a,14a), 69.22 (C-1,3,8,10), 77.87 (C-14c,14d), 124.37 (C-14′ ,14′′), 146.97 (C-13′,13′′), 175.83 (C-24′,24′′), 178.60 (C-23′,23′′), 179.23 (C-18′,18 ′′).

MALDI TOF/TOF: m/z (%) = 1210.1363 [ M - H] ⁺ (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 obtaining terpenoid dimer with perhydrotetracene spacer (1):

characterized in that cis- 1,6,7,12-tetraazaperhydrotetracene is reacted with N,N,N',N' -tetramethylmethanediamine and imidoamine MEMPK in CH ₃ OH in the presence of NiCl ₂ ⋅6H ₂ O catalyst at a molar ratio of cis- 1,6,7,12-tetraazaperhydrotetracene : N,N,N',N' -tetramethylmethanediamine : imidoamine MEMPK : NiCl ₂ ⋅6H ₂ O = 1 : 4 : 2 : (0.03-0.07) at room temperature (~20°C) temperature and atmospheric pressure for 2.5-3.5 hours.