Quinazoline small molecule compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a quinazoline small molecule compound as well as a preparation method and application thereof. The molecular formula of the quinazoline small molecule compound is C25H19N3O, the structural formula is # imgabs0, and the quinazoline small molecule compound is obtained by reacting 6-methoxy-2-naphthaldehyde and 2-(2-aminophenyl-)-1H-benzimidazole under a heating condition. The quinazoline small-molecule compound shows good selective inhibition on PTP1B, has a strong inhibition effect on HepG2 liver cancer cells at the cellular level, and can be applied to preparation of tumor drugs.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a quinazoline small molecule compound and a preparation method and application thereof. Background Art

[0002] Protein tyrosine phosphatases (PTPs) are non-metal enzymes widely present in cells. They regulate the phosphorylation level of intracellular tyrosine together with protein tyrosine kinases. They are expressed in various tissues of the human body and play an important role in various physiological processes. Once PTPs are expressed abnormally, it can cause excessive or insufficient tyrosine dephosphorylation, leading to many diseases related to signal transduction disorders, including cancer, diabetes, obesity, immune disorders, etc. At present, a large number of genetic and pharmacological studies have shown that a variety of PTPs are closely related to human diseases, and some of them are expected to become therapeutic targets for human diseases.

[0003] Among the huge family of PTPs, the research and application of PTP1B are the most eye-catching. PTP1B was isolated from human placental tissue in 1988 and was the first PTP to be successfully isolated. Its crystal structure was confirmed for the first time in 1994. PTP1B contains 435 amino acid residues, of which 30-278 amino acids constitute the catalytic functional region of the enzyme, and is located on the endoplasmic reticulum of the cell through the C-terminus of 35 proline residues. The main structural features of PTP1B are three highly conserved ring structures, namely the catalytic active region containing Cys215, the WPD ring and the second binding site. Recent literature reports that PTP1B is overexpressed in breast cancer and liver cancer tissues, and inhibitors of PTP1B may become potential therapeutic drugs.

[0004] At present, although a variety of PTP1B small molecule inhibitors have been reported, due to the high homology of the PTPs family, especially the 74% homology between TCPTP and PTP1B in sequence, there is still a lack of inhibitors with high activity and selectivity. Therefore, the development of highly effective and specific inhibitors for the treatment of PTP1B-related diseases is still of practical significance. Summary of the invention

[0005] In view of this, the purpose of the present invention is to overcome the deficiencies of the above-mentioned prior art and provide a quinazoline small molecule compound and a preparation method and application thereof.

[0006] To achieve the above object, the technical solution of the present invention is as follows:

[0007] A quinazoline small molecule compound, the molecular formula of which is: C 25 H 19 N3O, its structural formula is:

[0008]

[0009] The quinazoline small molecule compound belongs to the monoclinic crystal system, the space group is P21 / n, and the unit cell parameters are: α=90°, β=94.986(5), γ=90°.

[0010] The method for preparing the quinazoline small molecule compound as described above comprises the following steps: dissolving 6-methoxy-2-naphthaldehyde and 2-(2-aminophenyl-)-1H-benzimidazole in methanol, and heating and refluxing to obtain the quinazoline small molecule compound.

[0011] Furthermore, the molar ratio of the 6-methoxy-2-naphthaldehyde to the 2-(2-aminophenyl)-1H-benzimidazole is 1:1.

[0012] Furthermore, the heating reflux temperature is 60° C. and the time is 3 hours.

[0013] The asymmetric unit of the quinazoline small molecule compound of the present invention comprises one molecule. During the preparation process, under the action of heating, 6-methoxy-2-naphthaldehyde and 2-(2-aminophenyl-)-1H-benzimidazole undergo cyclization, and the ligand is converted from a Schiff base compound into a 6-(6-methoxynaphthalen-2-yl)-12,12a-dihydrobenzo[4,5]imidazo[1,2-c]quinazoline small molecule compound. Meanwhile, C—H···N in one molecule forms a weak molecular interaction with C—H···N in another molecule, thereby forming a one-dimensional chain structure.

[0014] The use of the quinazoline small molecule compound as described above as a PTP1B inhibitor.

[0015] A PTP1B inhibitor comprises the above-mentioned quinazoline small molecule compound.

[0016] Use of the quinazoline small molecule compound or PTP1B inhibitor as described above in the preparation of tumor drugs.

[0017] Furthermore, the tumor includes liver cancer.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The quinazoline small molecule compound of the invention is obtained by reacting 6-methoxy-2-naphthaldehyde with 2-(2-aminophenyl-)-1H-benzimidazole under heating conditions. The preparation method is simple, the product has high purity and a crystal structure is obtained, and the yield is high, which can reach 70%.

[0020] The quinazoline small molecule compound provided by the present invention can effectively inhibit PTP1B, and has a selectivity of about 5 times compared with the homologous TCPTP, and has a strong inhibitory effect on HepG2 human liver cancer cells. The quinazoline small molecule compound of the present invention will also be used as a potential candidate for treating PTP1B-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Synthesis route of the quinazoline small molecule compound of the present invention

[0022] Figure 2 Asymmetric structural unit of quinazoline small molecule compound of the present invention

[0023] Figure 3 One-dimensional chain diagram of the quinazoline small molecule compound of the present invention

[0024] Figure 4 Electrospray mass spectrometry of the quinazoline small molecule compound of the present invention

[0025] Figure 5 The IC of the quinazoline small molecule compound of the present invention for inhibiting PTP1B activity 50 Value determination curve

[0026] Figure 6 The IC of the quinazoline small molecule compound of the present invention for inhibiting TCPTP activity 50 Value determination curve

[0027] Figure 7 Effects of the quinazoline small molecule compounds of the present invention on the proliferation of HepG2 cells DETAILED DESCRIPTION

[0028] In order to facilitate the understanding of the present invention, the present invention will be described more fully below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0029] Example 1 Preparation of quinazoline small molecule compound

[0030] The synthetic route of quinazoline small molecule compounds is as follows Figure 1As shown, the specific process is as follows: weigh 6-methoxy-2-naphthaldehyde (0.186g, 1mmol) and add it to 10mL methanol, heat and stir until completely dissolved, then weigh 2-(2-aminophenyl-)-1H-benzimidazole (0.209g, 1mmol) and dissolve it in 10mL methanol, dissolve the latter and add it dropwise to the solution of the former, start stirring, heat and reflux at 60°C for 3h to form a colorless mixture solution. After the colorless mixture is cooled to room temperature, filter the mixture, put the filtrate into a clean glass vial, and let it stand at room temperature. After about 10 days, colorless block crystals precipitate on the wall of the bottle, and the quinazoline small molecule compound is obtained, with a yield of 72%. Elemental analysis C 25 H 19 N3O: Theoretical value: C, 79.55, H, 5.07, N, 11.13; Experimen...

[0031] Example 2 Crystal structure analysis of quinazoline small molecule compound

[0032] After trying a variety of methods to grow crystals, we finally obtained single crystals of quinazoline small molecule compounds by volatilization at room temperature. The structure of the compound was tested by selecting regular-shaped, relatively transparent block single crystals and sticking them on glass fibers under a microscope, and then testing them using a Bruker APEX-Ⅱ CCD diffractometer at the 3W1A beamline of Beijing Synchrotron Radiation Facility (BSRF). A graphite monochromator was used. X-ray diffraction data were collected at room temperature in an ω-2θ scanning mode, the unit cell parameters were determined by the SMART program, and the original data were restored and corrected by the SAINTPLUS program. The final data were analyzed by the direct method using the SHELXS-97 program package. Finally, the SHELXL197 (Sheldrick, 2008) program package was used to refine it, and the unit cell parameters and hydrogen bond information obtained are shown in Tables 1 and 2. From the crystal analysis results, it can be seen that the small molecule compound prepared by the present invention belongs to the monoclinic system, the space group is P21 / n, and the unit cell parameters are: α=90°,β=94.986(5),γ=90°. Its asymmetric unit contains a molecule (such as Figure 2 ), while the weak interactions between molecules form a one-dimensional chain structure (such as Figure 3 ).

[0033] Table 1 Crystallographic data of quinazoline small molecule compounds

[0034]

[0035] Table 2 Hydrogen bonding information of quinazoline small molecule compounds

[0036] D—H···A D—H H···A D.··A D—H···A <![CDATA[C14—H14···N1 i ]]> 0.98 2.60 3.569(4) 169

[0037] Example 3 Electrospray mass spectrometry test of quinazoline small molecule compounds

[0038] In order to study the existence form of the quinazoline small molecule compound of the present invention in solution, a small amount of compound crystals were dissolved in DMSO, the supernatant was taken and loaded into an electrospray mass spectrometer, and an electrospray ion source was used to detect and record data in a positive ion mode. Figure 4 Table 3 is the positive ion electrospray mass spectrum of the compound. The experimental values ​​are consistent with the theoretical values, indicating that the compound exists stably in the synthesized form.

[0039] Table 3 Electrospray mass spectra of quinazoline derivatives

[0040] Quinazoline small molecule compounds Species Experimental value Theoretical value <h2 style=";text-align:left;direction:ltr"><![CDATA[C <h2 style=";text-align:left;direction:ltr"> 25 <h2 style=";text-align:left;direction:ltr"> H<h2 style=";text-align:left;direction:ltr"> 19 <h2 style=";text-align:left;direction:ltr"> N3O]]><h2 style=";text-align:left;direction:ltr"> <![CDATA[[(1)+H] + ]]> 378.07 378.45

[0041] Example 4 IC of Quinazoline Small Molecule Compounds for Inhibition of PTP1B and TCPTP Activity 50 Value determination

[0042] IC 50 Determination method: The inhibitory effect of PTP1B and TCPTP activity was determined using the previously reported method, with p-nitrophenol phosphate (pNPP) as the substrate for the enzyme inhibition experiment. The inhibitor was dissolved in DMSO and diluted step by step. The experiment was carried out in a transparent 96-well plate. First, 83 μL of enzyme-containing MOPS buffer solution (pH = 7.24) was added to the first three rows of the ELISA plate. At the same time, a group of MOPS buffer solution without enzyme was added as a blank control group. 10 μL of DMSO solution was added to the first column, and the second to eighth columns were added as experimental groups with inhibitors of various concentration gradients (from small to large concentrations). After adding, they were fully mixed. After being placed in a 37°C constant temperature water bath for 30 minutes, 2 μL of 0.1M pNPP solution was added to start the reaction. After about 10 minutes, the solution in the 96-well plate turned yellow, and then 5 μL of 2M NaOH was added to terminate the reaction. The absorbance value of substrate decomposition (λ = 405nm) was measured, and finally the data was processed to obtain IC 50 The inhibitor solutions of each concentration gradient were freshly prepared before each experiment. This experiment needs to be repeated three times or more to ensure the authenticity and reliability of the experimental data. Figure 5 and 6 The half inhibitory concentration (IC50) of the quinazoline small molecule compound of the present invention on PTP1B 50 ) is: 0.69 μM, the half inhibitory concentration (IC 50 ) is: 3.20μM, which is a highly efficient and selective inhibitor.

[0043] Example 5 Study on the effect of quinazoline small molecule compounds on HepG2 cell proliferation

[0044] It was detected that PTP1B was overexpressed in HepG2 cells. In order to explore whether quinazoline small molecule compounds have an inhibitory effect on PTP1B in cells, HepG2 cells were selected for exploration. HepG2 cells were cultured in DMEM medium. When the cells grew to about 80%, the old medium was discarded, washed twice with 0.85% NaCl, the remaining liquid was sucked out with a pipette, an appropriate amount of trypsin was added to digest the cells, the supernatant was aspirated after centrifugation, 1 mL of fresh medium was added and aspirated evenly, an appropriate amount of cell suspension was added to the culture bottle containing 5-6 mL of culture medium, the "∞" shape was inverted and shaken well, and then placed in the incubator for culture.

[0045] When the cells grow to the logarithmic growth phase, wash them twice with 0.85% NaCl, add trypsin digestion, centrifuge, and add 1mL of fresh culture medium to make a cell suspension. According to the different culture time, inoculate 50μL of cell suspension of different concentrations into each well of the 96-well plate. After the cells adhere to the wall, add quinazoline small molecule compounds with final concentrations of 0, 1, 5, 10, 25, and 50μM for 24h, 48h, and 72h, add MTT and incubate for 4h, discard the supernatant, add 150μLDMSO and shake for about 10min, and then test the absorbance value with a microplate reader. The experimental results are shown in the figure. Figure 7 As shown in the figure, quinazoline small molecule compounds can significantly reduce the survival rate of HepG2 cells, and the cell survival rate is positively correlated with the action time and concentration. When 50μM quinazoline small molecule compounds acted on HepG2 cells for 24h, 48h, and 72h, the cell survival rates were 87.35%, 67.73%, and 45.61%, respectively. It is preliminarily speculated that quinazoline small molecule compounds form hydrogen bonds with certain proteins in cells, enhance the interaction between the complex and the protein, and thus show better anti-tumor activity.

[0046] The above description is only for better explaining the embodiments of the present invention, and is not intended to limit the present invention. Any modification or equivalent substitution that does not depart from the spirit and scope of the present invention shall fall within the scope of the present invention.

Claims

1. A quinazoline small molecule compound, characterized in that: Its molecular formula is: C 25 H 19 N3O, the structural formula is:

2. The quinazoline small molecule compound according to claim 1, characterized in that The quinazoline small molecule compound belongs to the monoclinic crystal system, the space group is P21 / n, and the unit cell parameters are: α=90°, β=94.986(5), γ=90°.

3. The method for preparing the quinazoline small molecule compound according to claim 1 or 2, characterized in that: The following steps are involved: 6-methoxy-2-naphthaldehyde and 2-(2-aminophenyl-)-1H-benzimidazole are dissolved in methanol, and heated under reflux to obtain the quinazoline small molecule compound.

4. The method for preparing a quinazoline small molecule compound according to claim 3, wherein The molar ratio of the 6-methoxy-2-naphthaldehyde to the 2-(2-aminophenyl)-1H-benzimidazole is 1:

1.

5. The method for preparing a quinazoline small molecule compound according to claim 3, characterized in that: The heating reflux temperature is 60° C. and the time is 3 h.

6. Use of the quinazoline small molecule compound according to claim 1 or 2 as a PTP1B inhibitor.

7. A PTP1B inhibitor, characterized in that Including the quinazoline small molecule compound according to claim 1 or 2.

8. Use of the quinazoline small molecule compound according to claim 1 or 2 or the PTP1B inhibitor according to claim 7 in the preparation of tumor drugs.

9. The use according to claim 8, characterized in that: The tumor includes liver cancer.

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