A diazatricyclic amine compound, preparation method and application thereof

Abstract

The present invention provides a diazatricyclic amine compound, a preparation method and an application thereof, and belongs to the field of tricyclic amine compounds. The diazatricyclic amine compound has a structure as shown in the following formula: #imgabs0#; wherein R is an alkyl group containing 1-6 carbon atoms, or R is M-(CH2) m R 1 ; M is CHJ or G, J is H or an alkyl group of 1-6 carbon atoms, and G is selected from a fully saturated or partially saturated cycloalkyl group; R 1 C(=NR 2 )NH、NHC(=NR 3 )NH2 or NR 4 R 5 ; said R 2 , R 3 , R 4 , R 5 are all independent alkyl groups with 1-4 carbon atoms, and m is an integer of 0-4. The results of biological activity tests on the diazatricyclic amine compound of the present invention show that the compound has an inhibitory effect on colon cancer cells.

Description

Technical Field

[0001] The present invention relates to the field of tricyclic amine compounds, and in particular to a diazatricyclic amine compound, a preparation method and application thereof. Background Art

[0002] DNA topoisomerase has been a hot topic in molecular biology research in recent years. Experiments have shown that it is directly related to cell proliferation, DNA replication, gene expression, and the study of anticancer drugs. Many drugs, such as doxorubicin and camptothecin, exert their cytotoxicity by affecting DNA topoisomerase activity. Therefore, topoisomerase has become a new target for screening anticancer drugs.

[0003] The benzonaphthyridine derivative N-[2-(dimethylamino)ethyl]-2,6-dimethyl-1-oxo-1,2-dihydrobenzo[b]-1,6-naphthyridine-4-carboxamide (SN28049) is a compound with a unique chemical structure and biological activity, belonging to the benzonaphthyridine class. Its complex mechanism of action may include toxicity to topo IIα, inhibition of topo I, and repression of gene transcription from promoters containing the SP-1 site. These effects may contribute to its potential anticancer activity in experimental solid tumors. SN28049 selectively binds to GC-rich DNA, interfering with normal DNA replication and transcription, thereby affecting cell growth and division. Several anticancer drugs, including doxorubicin and mitoxantrone, have a structure consisting of a DNA-intercalating chromophore linked to a positively charged side chain. The chromophore of SN28049 differs from earlier DNA-binding drugs in the presence of a methyl-substituted cyclic amide, a structural feature that may contribute to its anti-solid tumor activity.

[0004] Another tricyclic compound, DACA, and phenazine-1-carboxamide derivatives are active against the DNA regulatory enzymes topoisomerases I and II, and both exhibit cytotoxic effects. However, the relationship between the chemical structure of these compounds and their potential as anticancer drugs is not yet fully understood.

[0005] During the research on DACA, the inventors found that the substituent connected to the acridine ring has the general formula -CONH(CH2) n NR1R2, when R1 and R2 are both methyl groups, the compound has the highest activity. SN28049 also retains this structural feature and also exhibits extremely strong activity, which further illustrates that this structural feature helps to improve the activity of this type of compound.

[0006] The inventors discovered that all three compounds exhibited moderate biological activity, sharing the following common structural features: ① they all possessed aromatic three-membered ring structures; ② they all contained nitrogen atoms within the three-membered rings; and ③ the terminal nitrogen atoms of the substituents were all attached to dimethyl groups. Furthermore, existing research has shown that modifications to the substituents of SN28049 have significantly reduced their activity, with the difference in molecular structure between the modified and unmodified compounds exceeding four orders of magnitude (17,000:2). This suggests that the influence of the aromatic three-membered ring structure on the activity of these compounds is not limited to the substituents; the substituents are equally important. Furthermore, while SN28049 has demonstrated high activity in animal studies, it remains a significant step away from clinical application.

[0007] Thus, a tricyclic amine compound with a novel structure is provided, which can avoid the influence of the aromatic three-membered ring and substituents on its activity and obtain good biological activity, which has important technical significance and research value. Summary of the Invention

[0008] In order to solve the technical problems existing in the prior art, the present invention provides a diazatricyclic amine compound. By designing the molecular structure of the tricyclic amine compound, the influence of structural modification on its activity is avoided, thereby obtaining good biological activity. The present invention also provides a preparation method and application of the diazatricyclic amine compound.

[0009] In order to solve the above technical problems, the technical solutions adopted by the present invention are as follows:

[0010] A diazatricyclic amine compound, the molecular structure of which is shown in Formula I:

[0011] (I).

[0012] In formula (I), R is an alkyl group containing 1 to 6 carbon atoms, or R is M-(CH2) m R 1 ; M is CHJ or G, J is H or an alkyl group of 1-6 carbons, G is selected from a fully saturated or partially saturated cycloalkyl group; R 1 C(=NR 2 )NH、NHC(=NR 3 )NH2 or NR 4 R 5 ; the R 2 、R 3 、R 4 、R 5 Each is an independent alkyl group with 1 to 4 carbon atoms, and m is an integer of 0 to 4.

[0013] Preferably, the molecular structure of the diazatricyclic amine compound is as shown in Formula II:

[0014] (II).

[0015] A method for preparing a diazatricyclic amine compound, comprising: preparing an intermediate B, preparing an intermediate C, and preparing a diazatricyclic amine compound;

[0016] The method for preparing intermediate B is as follows: the starting material A is converted into an acyl chloride compound using thionyl chloride, and then reacted with aqueous ammonia to obtain intermediate B;

[0017] The structure of the starting material A is shown in Formula III:

[0018] (III);

[0019] The molecular structure of the intermediate B is shown in Formula IV:

[0020] (IV);

[0021] The method for preparing intermediate C is as follows: intermediate B is reduced with sodium borohydride to obtain intermediate C;

[0022] The molecular structure of the intermediate C is shown in Formula V:

[0023] (V);

[0024] The method for preparing the diazatricyclic amine compound is to condense the intermediate C with N,N-dimethylglycine or a salt thereof to obtain the diazatricyclic amine compound;

[0025] Preferably, in the preparation of intermediate B, the molar ratio of thionyl chloride to starting material A is 5-10:1.

[0026] Preferably, in the preparation of intermediate B, the molar ratio of ammonia water to starting material A is 2-5:1.

[0027] Preferably, in the preparation of intermediate C, the molar ratio of sodium borohydride to intermediate B is 3-6:1.

[0028] Preferably, in the preparation of the diazatricyclic amine compound, the molar ratio of N,N-dimethylglycine or its salt to the intermediate C is 1-5:1.

[0029] A use of the aforementioned diazatricyclic amine compound in the preparation of an anti-tumor drug; or a use of the aforementioned diazatricyclic amine compound as an intermediate in the preparation of an anti-tumor drug.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] (1) The diazatricyclic amine compound of the present invention can effectively avoid the influence of the substituent on the activity of the compound after the substituent is modified through specific molecular structure design, and the diazatricyclic amine compound has extremely strong biological activity; the preparation method of the diazatricyclic amine compound has a simple process flow, mild reaction conditions, easy control of the preparation process, low difficulty of post-processing, and is conducive to large-scale production.

[0032] (2) In the in vitro cytotoxicity test of the diazatricyclic amine compound of the present invention, the MTT inhibition rate data of compound Ia was calculated based on the MTT formazan generation state through the MTT experiment of HCT116 cells + diazatricyclic amine compound (Ia). The MTT inhibition rate data IC of compound Ia was 50 By the aforementioned biological activity test of diazatricyclic amine compounds, it can be seen that the IC of the diazatricyclic amine compound Ia of the present invention against HCT 116 human colon cancer cell line 50 0.8x10 -9 M; diazatricyclic amine compound Ia compared with SN 28049 (the IC of SN 28049 reported in the literature against HCT 116 human colon cancer cell line 50 8.4x10 -9 M), the activity is significantly improved, and it can be used as an active ingredient or intermediate in the preparation of anti-tumor drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is the hydrogen nuclear magnetic resonance spectrum of the diazatricyclic amine compound (i.e., Compound Ia) prepared in Example 1 of the present invention.

[0034] Figure 2 The figure shows the cytotoxicity test results of the diazatricyclic amine compound (ie, compound Ia) of Example 1 in the test example. In the figure, the left figure is the original inhibition rate curve, and the right figure is the log fitting curve of the inhibition rate. DETAILED DESCRIPTION

[0035] In order to have a clearer understanding of the technical features, purposes and effects of the present invention, the specific embodiments of the present invention are now described.

[0036] The present invention provides a diazatricyclic amine compound, the molecular structure of which is shown in Formula I:

[0037] (I).

[0038] In formula (I), R is an alkyl group containing 1 to 6 carbon atoms, or R is M-(CH2) m R 1 ; M is CHJ or G, J is H or an alkyl group of 1-6 carbons, G is selected from a fully saturated or partially saturated cycloalkyl group; R1 C(=NR 2 )NH、NHC(=NR 3 )NH2 or NR 4 R 5 ; the R 2 、R 3 、R 4 、R 5 Each is an independent alkyl group with 1 to 4 carbon atoms, and m is an integer of 0 to 4.

[0039] Preferably, the present invention provides a diazatricyclic amine compound, the molecular structure of which is shown in Formula II:

[0040] (II).

[0041] The present invention also provides a method for preparing a diazatricyclic amine compound, comprising: preparing an intermediate B, preparing an intermediate C, and preparing a diazatricyclic amine compound;

[0042] The method for preparing intermediate B is as follows: the starting material A is converted into an acyl chloride compound using thionyl chloride, and then reacted with aqueous ammonia to obtain intermediate B;

[0043] The structure of the starting material A is shown in Formula III:

[0044] (III);

[0045] The starting material A used in the embodiments of the present invention can be prepared by a preparation method disclosed in the prior art.

[0046] The molecular structure of the intermediate B is shown in Formula IV:

[0047] (IV);

[0048] The method for preparing intermediate C is as follows: intermediate B is reduced with sodium borohydride to obtain intermediate C;

[0049] The molecular structure of the intermediate C is shown in Formula V:

[0050] (V);

[0051] The method for preparing the diazatricyclic amine compound comprises the following steps: intermediate C is subjected to a condensation reaction with N,N-dimethylglycine or a salt thereof to obtain the diazatricyclic amine compound.

[0052] Furthermore, the method for preparing intermediate B is as follows: in a solvent environment, starting material A reacts with thionyl chloride under reflux, and then the solvent is removed; and then reacts with aqueous ammonia to obtain intermediate B.

[0053] Furthermore, the method for preparing intermediate C is to reduce intermediate B with sodium borohydride in a solvent environment in the presence of a boron trifluoride ether adduct solution at room temperature to obtain intermediate C.

[0054] Furthermore, the method for preparing the diazatricyclic amine compound is to react the intermediate C with N,N-dimethylglycine or its salt at room temperature in a solvent environment in the presence of 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride to obtain the diazatricyclic amine compound.

[0055] The preparation method of the diazatricyclic amine compound has a synthetic route as follows:

[0056] .

[0057] Preferably, in the preparation of intermediate B, the molar ratio of thionyl chloride to starting material A is 5-10:1, more preferably 5-6:1.

[0058] Preferably, in the preparation of intermediate B, the molar ratio of ammonia water to starting material A is 2-5:1, more preferably 4-5:1.

[0059] Preferably, in the preparation of intermediate C, the molar ratio of sodium borohydride to intermediate B is 3-6:1, more preferably 3.5-4.5:1.

[0060] Preferably, in the preparation of the diazatricyclic amine compound, the molar ratio of N,N-dimethylglycine or its salt to the intermediate C is 1-5:1, more preferably 1.2-2:1.

[0061] The present invention also provides a use of the aforementioned diazatricyclic amine compound in the preparation of an anti-tumor drug; or a use of the aforementioned diazatricyclic amine compound as an intermediate in the preparation of an anti-tumor drug.

[0062] The present invention will be further described below with reference to some specific embodiments.

[0063] Example 1

[0064] This embodiment provides a diazatricyclic amine compound Ia, the molecular structure of which is shown below:

[0065] .

[0066] This embodiment also provides a method for preparing a diazatricyclic amine compound Ia, and the specific steps are as follows:

[0067] 1. Preparation of intermediate B

[0068] The starting material A (0.27 g, 1.0 mmol), 10 mL of N,N-dimethylformamide (DMF), and thionyl chloride (SOCl2) (0.595 g, 5 mmol) were added to a four-necked flask, heated to 80°C, and refluxed for 2 h. After the reaction was completed as monitored by TLC, the solvent was removed by rotary evaporation, and ammonia water (concentration 33%, 0.175 g, 5 mmol), 10 mL of dichloromethane (CH2Cl2), and 10 mL of deionized water were added to the residue, and the reaction was carried out at 0°C for 2 h. After the reaction was completed as monitored by TLC, the reaction solution was extracted with dichloromethane (CH2Cl2) (25 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (eluent: pure ethyl acetate) to obtain 0.1 g of a yellow solid (i.e., intermediate B) with a yield of 37%.

[0069] H NMR spectrum of the obtained intermediate B:

[0070] 1 H NMR (400 MHz, CDCl3) δ 9.35(s,1H),7.93(d,J=8.4Hz,1H),7.78(d,J=6.0Hz,1H),7.57-7.53(m,1H),7.27(s,1H),3.74(s,3H),2.83(s,3H).

[0071] The starting material A used in this example was prepared by a preparation method disclosed in the prior art. The molecular structure of the starting material A is shown below:

[0072] .

[0073] 2. Preparation of intermediate C

[0074] To a four-necked flask, add intermediate B (27 mg, 0.1 mmol) and 5 mL of tetrahydrofuran, mix and dissolve, then add sodium borohydride (15 mg, 0.4 mmol) and boron trifluoride ether adduct solution (40 mg, 0.28 mmol), and stir at room temperature overnight. After the reaction is completed as monitored by TLC, add 2 mL of saturated aqueous ammonium chloride solution, stir for 15 min, and concentrate to remove tetrahydrofuran. After extraction with dichloromethane, dry and concentrate to obtain a yellow solid (i.e., intermediate C), which is directly used in the next reaction.

[0075] 3. Preparation of diazatricyclic amine compound Ia

[0076] To a four-necked flask, N,N-dimethylglycine hydrochloride (19 mg, 0.12 mmol) and 2 mL of N,N-dimethylformamide (DMF) were added and fully dissolved. 1-Hydroxybenzotriazole (HOBt) (18 mg, 0.12 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) (23 mg, 0.12 mmol) were then added and stirred at room temperature for 30 min. Then, the above-mentioned intermediate C (25 mg, 0.1 mmol) was added and stirred at room temperature overnight. After completion of the reaction as monitored by TLC, the reaction solution was poured into 2 mL of water. Solid precipitated, which was filtered and dried to obtain 21 mg of a yellow solid (i.e., diazatricyclic amine compound Ia) in a yield of 62%.

[0077] like Figure 1 As shown, the nuclear magnetic resonance hydrogen spectrum of the diazatricyclic amine compound Ia prepared in this example is:

[0078] 1 H NMR (400 MHz, CDCl3) δ11.13(s,1H),9.33(t,1H),8.63(d,1H),7.94(d,1H),7.77(d,1H),7.56(dd,1H),3.74-3.50(m,5H),2.85(m,3H),2.45(m,2H),2.28(s,6H).

[0079] This embodiment also provides the use of the diazatricyclic amine compound Ia in the preparation of anti-tumor drugs; or the use of the diazatricyclic amine compound Ia as an intermediate in the preparation of anti-tumor drugs.

[0080] Test Example 1

[0081] Biological tests on diazatricyclic amine compounds

[0082] The diazatricyclic amine compound of the present invention was subjected to an in vitro cytotoxicity test by using an MTT test of diazatricyclic amine compound + HCT116 cells. The drug preparation method is as follows:

[0083] Drug stock solution: Dissolve 1 mg of compound Ia in 500 μL of DMSO. Mix the solution thoroughly and add 6.4 μL of the drug solution to 4.994 mL of DMEM-H medium (5 mL volume). The drug concentration is 2.56 μg / mL. This solution is the stock solution. Sterilize the drug by filtering it through a 0.22 μm filter.

[0084] Drug sub-solution: 62.5 μl of the drug stock solution of compound Ia was added to 1.938 mL of DMEM-H culture medium and mixed evenly (preparation volume 2 mL). The drug concentration at this point was 80 ng / mL, and this drug solution was the sub-solution.

[0085] A DMSO control group was set up by adding 0.5 mL of DMSO to 4.5 mL of DMEM-H medium and mixing well to prepare a 10% DMSO control group.

[0086] Finally, the MTT inhibition rate data was calculated based on the MTT formazan generation status. The MTT inhibition rate data IC of compound Ia was 50 It is 270ng / mL.

[0087] According to the biological activity test results of the above-mentioned diazatricyclic amine compounds (such as Figure 2 As shown), it can be seen that the IC of the diazatricyclic amine compound Ia of the present invention against the HCT 116 human colon cancer cell line is 50 0.8x10 -9 M.

[0088] Unless otherwise specified, all percentages used in the present invention are by mass.

[0089] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art will be able to modify the technical solutions described in the aforementioned embodiments or substitute equivalents for some of the technical features. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A diazatricyclic amine compound, characterized in that The molecular structure is shown in formula II: (II)。 2. A method for preparing a diazatricyclic amine compound, characterized in that: Including: preparing intermediate B, preparing intermediate C, preparing diazatricyclic amine compounds; The method for preparing intermediate B is that the starting material A is converted into an acyl chloride compound by using thionyl chloride, and then reacted with aqueous ammonia to obtain intermediate B; The molecular structure of the starting material A is shown in Formula III: (III); The molecular structure of the intermediate B is shown in Formula IV: (IV); The method for preparing intermediate C is that intermediate B is reduced by sodium borohydride to obtain intermediate C; The molecular structure of the intermediate C is shown in Formula V: （V）； The method for preparing the diazatricyclic amine compound is that the intermediate C is subjected to a condensation reaction with N,N-dimethylglycine or a salt thereof to obtain the diazatricyclic amine compound; The molecular structure of the diazatricyclic amine compound is shown below: 。 3. The method for preparing a diazatricyclic amine compound according to claim 2, characterized in that: In the preparation of intermediate B, the molar ratio of thionyl chloride to starting material A is 5-10:

1.

4. The method for preparing a diazatricyclic amine compound according to claim 2, characterized in that: In the preparation of intermediate B, the molar ratio of ammonia water to starting material A is 2-5:

1.

5. The method for preparing a diazatricyclic amine compound according to claim 2, characterized in that: In the preparation of intermediate C, the molar ratio of sodium borohydride to intermediate B is 3-6:

1.

6. The method for preparing a diazatricyclic amine compound according to claim 2, characterized in that: In the preparation of the diazatricyclic amine compound, the molar ratio of N,N-dimethylglycine or its salt to the intermediate C is 1-5:

1.

7. A use of the diazatricyclic amine compound as claimed in claim 1, characterized in that: The diazatricyclic amine compound is used in the preparation of anti-tumor drugs; the tumor is human colon cancer.

Images