A diazatricyclic amine compound, preparation method and application

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 compounds of the present invention show that the compounds have 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 is one of the hot topics 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 cytotoxicity by affecting the activity of DNA topoisomerase. Therefore, topoisomerase has now 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 of compounds; it has a complex mechanism of action that may include toxicity to topo IIα, inhibition of topo I, and inhibition of gene transcription containing SP-1 site promoters, which may contribute to its potential in experimental solid tumor anticancer activity. SN28049 can selectively bind to GC-rich DNA, interfere with normal DNA replication and transcription, and thus affect cell growth and division. Many anticancer drugs, including doxorubicin and mitoxantrone, have a structure consisting of a DNA-intercalated chromophore connected to a positively charged side chain. The chromophore of SN28049 differs from earlier DNA-binding drugs in that it has a methyl-substituted cyclic amide, a structural feature that may contribute to its anti-solid tumor activity to some extent.

[0004] Another tricyclic compound, DACA, and phenazine-1-carboxamide derivatives are active against the DNA regulatory enzymes topoisomerase 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 still unclear.

[0005] The inventors found in the process of studying DACA that the substituent connected to the acridine ring has the general formula -CONH(CH2) n NR 1 R 2 , when R 1 and R 2 When it is a methyl group, the compound has the highest activity. SN28049 also retains this structural feature and also shows extremely strong activity, which further illustrates that this structural feature helps to improve the activity of this type of compound.

[0006] The inventors found that the above three compounds all exhibit certain biological activity, and their common structural features are as follows: ① All are aromatic three-membered ring structures; ② All have N atoms in the three-membered ring; ③ All N atoms at the terminal of the substituent are connected to dimethyl. At the same time, scholars have modified the substituents of SN28049 in existing studies. Although there is only a slight difference between the molecular structure of the compound before and after the modification, the activity of the compound before and after the modification is greatly reduced, up to four orders of magnitude (17000:2). Based on this, it can be concluded that the influence on the activity of this type of compound is not limited to the aromatic three-membered ring structure of the compound, but the substituent is also equally important. At the same time, although SN28049 showed high activity in animal experiments, it is still a long way from entering the clinic.

[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, which avoids the influence of structural modification on its activity by designing the molecular structure of the tricyclic amine compound, 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 solution adopted by the present invention is 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-(CH 2 ) m R 1 ; M is CHJ or G, J is H or an alkyl group of 1-6 carbon atoms, G is selected from a fully saturated or partially saturated cycloalkyl group; R 1 C(=NR 2 )NH、NHC(=NR 3 )NH 2 or NR 4 R 5 ; said R 2 , R 3 , R 4 , R 5 Each is an independent alkyl group of 1 to 4 carbon atoms, and m is an integer of 0 to 4.

[0013] Preferably, the diazatricyclic amine compound has a molecular structure as shown in Formula II or Formula III:

[0014] (II);

[0015] (III).

[0016] A method for preparing a diazatricyclic amine compound, comprising: preparing an intermediate B, preparing an intermediate C, and preparing a diazatricyclic amine compound;

[0017] 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;

[0018] The structure of the starting material A is shown in Formula IV:

[0019] (IV);

[0020] The molecular structure of the intermediate B is shown in Formula V:

[0021] (V);

[0022] The method for preparing intermediate C is that intermediate B reacts with trichloroisocyanuric acid and then is treated with potassium hydroxide to obtain intermediate C;

[0023] The molecular structure of the intermediate C is shown in Formula VI:

[0024] (VI);

[0025] The method for preparing the diazatricyclic amine compound is to condense the intermediate C with an N,N-dimethylated amino acid or a salt thereof to obtain the diazatricyclic amine compound;

[0026] Preferably, the N,N-dimethylated amino acid is N,N-dimethylglycine or N,N-dimethyl-β-alanine.

[0027] Preferably, in the preparation of intermediate B, the molar ratio of thionyl chloride to starting material A is 5-10:1.

[0028] The molar ratio of aqueous ammonia to starting material A is 2-5:1.

[0029] Preferably, in the preparation of intermediate C, the molar ratio of trichloroisocyanuric acid to intermediate B is 1-4:1.

[0030] Preferably, in the preparation of the diazatricyclic amine compound, the molar ratio of the N,N-dimethylated amino acid or its salt to the intermediate C is 1-5:1.

[0031] 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.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] (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 a 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-treatment, and is conducive to large-scale production.

[0034] (2) In the in vitro cytotoxicity test of the diazatricyclic amine compounds of the present invention, the MTT inhibition rate data of compound Ia was calculated based on the MTT formazan generation state by performing an MTT experiment on HCT116 cells + diazatricyclic amine compounds (Ia, Ib). The MTT inhibition rate data IC of compound Ia was 50 The MTT inhibition rate of compound Ib is 429µg / mL. 50 The above-mentioned biological activity test of the diazatricyclic amine compounds shows that the IC value of the diazatricyclic amine compound Ia of the present invention against the HCT 116 human colon cancer cell line is 0.586 μg / mL. 50 1.26x10 -6 M; IC of diazatricyclic amine compound Ib against HCT 116 human colon cancer cell line 50 1.73x10 -9 M; Among them, the diazatricyclic amine compound Ib is 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 effective ingredient or intermediate in the preparation of anti-tumor drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] 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.

[0036] Figure 2 This is the hydrogen nuclear magnetic resonance spectrum of the diazatricyclic amine compound (i.e., compound Ib) prepared in Example 2 of the present invention.

[0037] Figure 3The 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 inhibition rate log fitting curve.

[0038] Figure 4 The cytotoxicity test results of the diazatricyclic amine compound (ie, compound Ib) of Example 2 in the test example; in the figure, the left figure is the original inhibition rate curve, and the right figure is the inhibition rate log fitting curve. DETAILED DESCRIPTION

[0039] In order to have a clearer understanding of the technical features, purposes and effects of the present invention, the specific implementation methods of the present invention are now described.

[0040] The present invention provides a diazatricyclic amine compound, the molecular structure of which is shown in Formula I:

[0041] (I).

[0042] In formula (I), R is an alkyl group containing 1 to 6 carbon atoms, or R is M-(CH 2 ) m R 1 ; M is CHJ or G, J is H or an alkyl group of 1-6 carbon atoms, G is selected from a fully saturated or partially saturated cycloalkyl group; R 1 C(=NR 2 )NH、NHC(=NR 3 )NH 2 or NR 4 R 5 ; said R 2 , R 3 , R 4 , R 5 Each is an independent alkyl group of 1 to 4 carbon atoms, and m is an integer of 0 to 4.

[0043] Preferably, the diazatricyclic amine compound has a molecular structure as shown in Formula II or Formula III:

[0044] (II);

[0045] (III).

[0046] 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;

[0047] 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;

[0048] The structure of the starting material A is shown in Formula IV:

[0049] (IV);

[0050] The starting material A used in the embodiments of the present invention can be prepared by a preparation method disclosed in the prior art.

[0051] The molecular structure of the intermediate B is shown in Formula V:

[0052] (V);

[0053] The method for preparing intermediate C is that intermediate B reacts with trichloroisocyanuric acid and then is treated with potassium hydroxide to obtain intermediate C;

[0054] The molecular structure of the intermediate C is shown in Formula VI:

[0055] (VI);

[0056] The method for preparing the diazatricyclic amine compound is to condense the intermediate C with an N,N-dimethylated amino acid or a salt thereof to obtain the diazatricyclic amine compound;

[0057] Preferably, the N,N-dimethylated amino acid is N,N-dimethylglycine or N,N-dimethyl-β-alanine.

[0058] Furthermore, the method for preparing intermediate B is: in a solvent environment, the starting material A reacts with thionyl chloride under reflux, and then the solvent is removed; and then the starting material A reacts with aqueous ammonia to obtain intermediate B.

[0059] Furthermore, the method for preparing intermediate C is that intermediate B reacts with trichloroisocyanuric acid at room temperature in a solvent environment to obtain intermediate C.

[0060] Furthermore, the method for preparing the diazatricyclic amine compound is to react the intermediate C with an N,N-dimethylated amino acid or a salt thereof 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.

[0061] The preparation method of the diazatricyclic amine compound has a synthetic route as follows:

[0062] .

[0063] 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.

[0064] The molar ratio of aqueous ammonia to starting material A is 2-5:1, more preferably 4-5:1.

[0065] Preferably, in the preparation of intermediate C, the molar ratio of trichloroisocyanuric acid to intermediate B is 1-4:1, more preferably 2.5-3.5:1.

[0066] Preferably, in the preparation of the diazatricyclic amine compound, the molar ratio of the N,N-dimethylated amino acid or its salt to the intermediate C is 1-5:1, more preferably 1.2-2:1.

[0067] The present invention also provides a use of the aforementioned diazatricyclic amine compound in the preparation of anti-tumor drugs; or the use of the aforementioned diazatricyclic amine compound as an intermediate in the preparation of anti-tumor drugs.

[0068] The present invention is further described below in conjunction with some specific embodiments.

[0069] Example 1

[0070] This embodiment provides a diazatricyclic amine compound Ia, the molecular structure of which is shown below:

[0071] .

[0072] This embodiment also provides a method for preparing a diazatricyclic amine compound Ia, and the specific steps are as follows:

[0073] 1. Preparation of intermediate B

[0074] Add the starting material A (0.27 g, 1.0 mmol), 10 mL of N,N-dimethylformamide (DMF), and thionyl chloride (SOCl) into a four-necked flask. 2 (0.595 g, 5 mmol), heated to 80 ° C, kept under reflux 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) and 10 mL of dichloromethane (CH 2 Cl 2 , 10 mL of deionized water, react at 0 °C for 2 h; after the reaction was completed, dichloromethane (CH 2 Cl 2 The reaction solution was extracted with 25 mL × 3, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography (eluent: pure ethyl acetate) to obtain 0.1 g of yellow solid (i.e., intermediate B), with a yield of 37%.

[0075] The hydrogen nuclear magnetic resonance spectrum of the obtained intermediate B:

[0076] 1H NMR (400 MHz, CDCl 3 ) 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).

[0077] The starting material A used in this example is prepared by a preparation method disclosed in the prior art, and the molecular structure of the starting material A is shown below:

[0078] .

[0079] 2. Preparation of intermediate C

[0080] Into a four-necked flask, intermediate B (27 mg, 0.1 mmol), trichloroisocyanuric acid TCCA (8.5 mg, 0.037 mmol), 3 mL of methanol MeOH, and 2 mL of dichloromethane CH 2 Cl 2 Dissolve and stir at room temperature overnight. After the reaction is completed as monitored by TLC, remove the solvent by rotary evaporation, add 50 mL of deionized water and 3 g of potassium hydroxide (KOH) to the residue, and reflux at 100°C for 3 h. After cooling to room temperature, solid precipitates. Filter to obtain a yellow solid (i.e., intermediate C), which is directly used in the next step.

[0081] 3. Preparation of diazatricyclic amine compound Ia

[0082] N,N-dimethyl-β-alanine hydrochloride (19 mg, 0.12 mmol) and 2 mL of N,N-dimethylformamide DMF were added to a four-necked flask to fully dissolve, and then 1-hydroxybenzotriazole HOBt (18 mg, 0.12 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride EDC·HCl (23 mg, 0.12 mmol), and N,N-diisopropylethylamine DIPEA (20 mg) were added, and stirred at room temperature for 30 min. Then, the above intermediate C (24 mg, 0.1 mmol) was added, and stirred at room temperature overnight. After the reaction was completed as monitored by TLC, the reaction solution was poured into 2 mL of deionized water, and solids were precipitated. The solids were filtered to obtain the solids, and dried to obtain 25 mg of yellow solids (i.e., diazatricyclic amine compound Ia), with a yield of 76%.

[0083] like Figure 1 As shown, the nuclear magnetic resonance hydrogen spectrum of the diazatricyclic amine compound Ia prepared in this example is:

[0084] 1 H NMR (400 MHz, CDCl3 ) δ 10.80(s,1H),9.29(s,1H),7.93-7.87(m,3H),7.28(s,1H),3.67(s,3H),3.54(s,3H),3.52-3.46(m,4H),2.95(s,3H),2.82(s,3H).

[0085] This embodiment also provides a use of a diazatricyclic amine compound Ia in the preparation of an anti-tumor drug; or a use of a diazatricyclic amine compound Ia as an intermediate in the preparation of an anti-tumor drug.

[0086] Example 2

[0087] This embodiment provides a diazatricyclic amine compound Ib, the molecular structure of which is shown below:

[0088] .

[0089] This embodiment also provides a method for preparing a diazatricyclic amine compound Ib, and the specific steps are as follows:

[0090] 1. Preparation of intermediate B

[0091] This step is the same as that in Example 1 and will not be repeated here.

[0092] 2. Preparation of intermediate C

[0093] This step is the same as that in Example 1 and will not be repeated here.

[0094] 3. Preparation of diazatricyclic amine compound Ib

[0095] N,N-dimethylglycine hydrochloride (19 mg, 0.12 mmol) and 2 mL of N,N-dimethylformamide DMF were added to a four-necked flask to fully dissolve, and then 1-hydroxybenzotriazole HOBt (18 mg, 0.12 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride EDC·HCl (23 mg, 0.12 mmol) were added, and stirred at room temperature for 30 min; then intermediate C (24 mg, 0.1 mmol) was added, and stirred at room temperature overnight; after the reaction was completed as monitored by TLC, the reaction solution was poured into 2 mL of deionized water, and solids were precipitated. The solids were filtered to obtain 21 mg of yellow solids (i.e., diazatricyclic amine compound Ib), and the yield was 65%.

[0096] like Figure 2 As shown, the nuclear magnetic resonance hydrogen spectrum of the diazatricyclic amine compound Ib prepared in this example is:

[0097] 1H NMR (400 MHz, CDCl 3 ) δ10.81(s,1H),9.28(s,1H),7.93-7.87(m,3H),7.28(s,1H),3.67(s,3H),3.54(s,3H),3.45(s,2H),2.95(s,3H),2.79(s,3H).

[0098] This embodiment also provides a use of a diazatricyclic amine compound Ib in the preparation of an anti-tumor drug; or a use of a diazatricyclic amine compound Ib as an intermediate in the preparation of an anti-tumor drug.

[0099] Test Example 1

[0100] Biological Tests of Diazatricyclic Amines

[0101] The diazatricyclic amine compound of the present invention was subjected to an in vitro cytotoxicity test by using the MTT test of diazatricyclic amine compound + HCT116 cells. The drug preparation method is as follows:

[0102] Drug mother solution: Dissolve 1 mg of compound Ia and compound Ib in 500 uL of DMSO respectively. After mixing the solution, take 6.4 uL of the drug solution and add it to 4.994 mL of DMEM-H culture medium (preparation volume 5 mL). At this time, the drug concentration is 2.56 ug / mL; this drug solution is the mother solution, and the drug is filtered and sterilized with a 0.22 um filter.

[0103] Drug sub-liquid: Take 62.5ul of the drug mother solution of compound Ia or compound Ib and add it to 1.938mL of DMEM-H culture medium and mix well (preparation volume 2mL). At this time, the drug concentration is 80ng / mL, and this drug solution is the sub-liquid.

[0104] 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.

[0105] 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 The MTT inhibition rate of compound Ib is 297µg / mL. 50 It is 0.586µg / mL.

[0106] According to the biological activity test results of the above-mentioned diazatricyclic amine compounds (such as Figure 3-4 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 0.92x10-6 M; IC of diazatricyclic amine compound Ib against HCT 116 human colon cancer cell line 50 1.73x10 -9 M.

[0107] Unless otherwise specified, all percentages used in the present invention are by mass.

[0108] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is 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 can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A diazatricyclic amine compound, characterized in that The molecular structure is shown in formula III: (III)。 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 IV: (IV); The molecular structure of the intermediate B is shown in Formula V: （V）； The method for preparing intermediate C is that intermediate B reacts with trichloroisocyanuric acid and then is treated with potassium hydroxide to obtain intermediate C; The molecular structure of the intermediate C is shown in Formula VI: (WE); 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 diazatricyclic amine compound has the following molecular structure: 。 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; The molar ratio of aqueous ammonia to starting material A is 2-5:

1.

4. 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 trichloroisocyanuric acid to intermediate B is 1-4:

1.

5. 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.

6. A use of the diazatricyclic amine compound as claimed in claim 1, characterized in that: Application of the diazatricyclic amine compound in the preparation of anti-tumor drugs; or application of the diazatricyclic amine compound as an intermediate in the preparation of anti-tumor drugs.

Images