An indole iodine salt-pyridine bis-hemicyanine compound, a preparation method and application thereof

Abstract

The application belongs to the field of tumor targeted therapy, and particularly relates to an indole iodine salt-pyridine double hemicyanine compound and a preparation method and application thereof. The structural formula of the indole iodine salt-pyridine double hemicyanine compound is shown in formula (I) or formula (II): two 3,3-dimethyl-3H-indole iodides are connected to the two sides of a pyridine nucleus through dimethyl chain, two compounds with high biological activity are synthesized, and the wide application of the compounds in biological medicine and the development prospect of pharmaceutical preparations are expanded. The compounds can significantly inhibit the proliferation of breast cancer, lung cancer and colorectal cancer cells at a lower dose, and have mitochondria targeting, which indicates that the compounds have the prospect of being developed into anti-tumor drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of tumor targeted therapy, and particularly relates to an indole iodine salt-pyridine bis-hemicyanine compound as well as a preparation method and application thereof. BACKGROUND

[0002] Cancer is an important cause of human death due to diseases worldwide. Because the early symptoms of these cancers are not specific, most patients are in the middle and late stages when diagnosed, the tumor metastasis and recurrence rate after surgery is high, and the side effects of traditional treatment methods such as chemotherapy and radiotherapy are large and are prone to drug resistance, and the clinical benefit rate is low. Therefore, it is of great significance to develop new targeted anti-cancer drugs.

[0003] Mitochondria are the main place to provide energy in cells, and are also the key regulators of various activities such as cell survival, metabolism, ion homeostasis, oxidation and reduction, and apoptosis. Studies have shown that mitochondrial abnormalities are related to the occurrence and development of cancer. The permeability of mitochondrial membrane can affect the apoptosis of cancer cells by regulating the activity of intracellular caspases. The production of reactive oxygen species in mitochondria can promote the proliferation, survival, angiogenesis and metastasis of cancer cells. Different strategies can be used to target the mitochondria of cancer cells according to the characteristics of the mitochondria of cancer cells different from those of normal cells, such as increased ROS levels, increased temperature, unbalanced membrane potential, and decreased matrix pH. Therefore, mitochondria are potential therapeutic targets for cancer.

[0004] Indole compounds belong to heterocyclic derivative alkaloids, and have good anti-inflammatory, analgesic, antiviral and antitumor activities. Indole compounds have been used to prepare various antitumor drugs such as vincristine (microtubule protein inhibitor), osimertinib mesylate (kinase inhibitor), goserelin acetate (gonadotropin-releasing hormone receptor (GnRHR) inhibitor), and paribranib (histone deacetylase inhibitor). Some preclinical studies have also shown that some new compounds synthesized with indole as the structural backbone have good potential in targeted anti-cancer research. Patent CN109400604A discloses 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole compounds and uses thereof. The compounds can be used as a multi-target receptor protein tyrosine kinase inhibitor to effectively inhibit the growth of tumor cells. It is not yet known whether these clinical drugs and preclinical compounds can target mitochondria. Therefore, the development of new indole compounds with mitochondrial targeting properties will provide new drug options for tumor targeted therapy. SUMMARY

[0005] To solve the above technical problems, the present application provides an indole iodine salt-pyridine bis-hemicyanine compound as well as a preparation method and application thereof.

[0006] To achieve the above-mentioned purposes, the technical scheme of the present application is as follows:

[0007] An indolium iodine salt-pyridine bis-hemicyanine compound has a structural formula as shown in formula (I) or formula (II):

[0008]

[0009] The preparation method of the above-mentioned indolium iodine salt-pyridine bis-hemicyanine compound is as follows: 1,2,3,3-tetramethyl-3H-indolium iodide and pyridine dimethyl formaldehyde are dissolved in a solvent, and reacted under argon protection, and the indolium iodine salt-pyridine bis-hemicyanine compound is obtained after the reaction is completed.

[0010] Further, the pyridine dimethyl formaldehyde is 2,6-pyridine dimethyl formaldehyde or 3,5-pyridine dimethyl formaldehyde.

[0011] Further, the solvent is ethanol, methanol or acetone.

[0012] Further, the molar ratio of the 1,2,3,3-tetramethyl-3H-indolium iodide to the pyridine dimethyl formaldehyde is (2.2-2.5):1.

[0013] Further, the concentration of the pyridine dimethyl formaldehyde is 0.017-0.024 mol / L.

[0014] Further, the reaction temperature is 70-100 DEG C, and the reaction time is 6-18 h.

[0015] Further, the post-treatment step after the reaction is completed is further included: when the reactant is 2,6-pyridine dimethyl formaldehyde, the solvent is removed by rotary evaporation and column chromatography is used for separation and purification; when the reactant is 3,5-pyridine dimethyl formaldehyde, the solvent is removed by rotary evaporation, washed by acetonitrile and dried.

[0016] The above-mentioned indolium iodine salt-pyridine bis-hemicyanine compound and biologically acceptable salts and preparations thereof are used in the preparation of cancer treatment drugs.

[0017] Further, the cancer refers to breast cancer, lung cancer and colorectal cancer.

[0018] The beneficial effects of the present application are as follows:

[0019] (1) The present application synthesizes a kind of indolium iodine salt-pyridine bis-hemicyanine compound DII-2,6-Py and DII-3,5-Py with a new structure. The proliferation inhibition effect of the compound on various cancer cells is detected by MTT method, and the targeting of the compound to mitochondria is detected by fluorescence imaging, which can be co-localized with mitochondrial specific dye.

[0020] (2) The compounds DII-2,6-Py and DII-3,5-Py of the present application can effectively inhibit the proliferation of cells of breast cancer, lung cancer, colorectal cancer and the like, and the drug efficacy is superior to that of 5-Fluorouracil (5Fu), a broad-spectrum anti-tumor chemotherapy drug. Specifically, the IC50 of 5Fu in MDA-MB-31, BT549, SW480 and A549 cell lines is about 17.9 times, 6.57 times, 1.79 times, 38.57 times and 4.5 times of that of DII-2,6-Py, and about 26.04 times, 11.43 times, 3.44 times, 20.08 times and 6.29 times of that of DII-3,5-Py, respectively, indicating that the sensitivity of DII-2,6-Py and DII-3,5-Py to tumor cells (especially breast cancer and lung cancer cells) is obviously superior to that of 5Fu, and the proliferation of these cells can be significantly inhibited. BRIEF DESCRIPTION OF DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or prior art description will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative effort on the basis of these drawings.

[0022] Figure 1 is the HR-ESI-MS spectrum of DII-2,6-Py.

[0023] Figure 2 is the HR-ESI-MS spectrum of DII-2,6-Py. 1 is the H NMR spectrum of DII-2,6-Py.

[0024] Figure 3 is the H NMR spectrum of DII-2,6-Py. 13 is the C NMR spectrum of DII-2,6-Py.

[0025] Figure 4 is the HR-ESI-MS spectrum of DII-3,5-Py.

[0026] Figure 5 is the HR-ESI-MS spectrum of DII-3,5-Py. 1 is the H NMR spectrum of DII-3,5-Py.

[0027] Figure 6 is the HR-ESI-MS spectrum of DII-3,5-Py.

[0028] Figure 7The proliferation inhibition activities of DII-2,6-Py and DII-3,5-Py on colorectal cancer cell lines SW480 and DLD-1 were characterized by MTT method with 5Fu as a positive control drug.

[0029] Figure 8 The proliferation inhibition activities of DII-2,6-Py and DII-3,5-Py on lung cancer cell lines A549 and H460 were characterized by MTT method with 5Fu as a positive control drug.

[0030] Figure 9 The mitochondrial co-localization experiments of DII-2,6-Py and DII-3,5-Py were performed by fluorescence imaging method. DETAILED DESCRIPTION

[0031] The technical solutions of the present application will be clearly and completely described below in combination with the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0032] In the method for synthesizing the compound of formula I in the present application, various raw materials used in the reaction can be prepared by those skilled in the art based on existing knowledge, or can be prepared by methods known from the literature, or can be commercially available. The intermediates, raw materials, reagents, reaction conditions, etc. used in the above reaction scheme can be appropriately changed based on the existing knowledge of those skilled in the art.

[0033] In the present application, unless otherwise specified, (i) the temperature is expressed in degrees Celsius (℃), and the operation is carried out at room temperature; more specifically, the room temperature refers to 20-30℃; (ii) the organic solvent is dried by a commonly used drying method, and the evaporation of the solvent uses a rotary evaporator for evaporation under reduced pressure, with a bath temperature not higher than 50℃; the developing agent and eluent are both by volume; (iii) the reaction process is tracked by thin layer chromatography (TLC); (iv) the final product has a satisfactory proton nuclear magnetic resonance (1H-NMR).

[0034] Example 1

[0035] The preparation method of the indole iodine salt-pyridine bis-hemicyanine compound in this embodiment is as follows:

[0036] 147.5 mg (1.1 mmol) of 2,6-pyridinedicarboxaldehyde and 758.5 ​​mg (2.5 mmol) of 1,2,3,3-tetramethyl-3H-indole iodide were weighed into a two-necked round-bottom flask, and 60 mL of ethanol was added. The mixture was reacted at 70 °C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain the crude product. The crude product was purified by column chromatography (dichloromethane-methanol, v / v 20:1) to obtain 517.5 mg of the brick-red solid DII-2,6-Py, as shown in structural formula (Ⅰ), with a yield of 68%.

[0037]

[0038] The compound shown in formula (Ⅰ) was subjected to HR-ESI-MS spectra, 1 H NMR spectrum and 13 The specific data from the C NMR spectroscopy test are shown below:

[0039] HR-ESI-MS m / z:calcd for C 31 H 33 I₂N₃, 478.2864[M⁻²I⁺CH₃O₂] - ] + 223.6337[M-2I] 2+ ;found478.3604[M-2I+CH3O - ] + 223.6650 [M-2I] 2+ The specific spectrum is as follows Figure 1 As shown.

[0040] 1 H-NMR (400MH) Z DMSO-d6) δ: 8.55 (d, J = 16.0 Hz, 2H), 8.31–8.27 (m, 5H), 8.07–8.05 (m, 2H), 7.98–7.95 (m, 2H), 7.73–7.70 (m, 4H), 4.29 (s, 6H), 1.86 (s, 12H), detailed spectrum as follows. Figure 2 As shown.

[0041] 13 C-NMR (100MH) Z DMSO-d6) δ: 181.7, 152.2, 148.8, 144.0, 141.9, 139.3, 130.2, 129.9, 129.2, 123.1, 117.2, 116.0, 52.7, 35.3, 24.9. Specific spectra are shown below. Figure 3 As shown.

[0042] Example 2

[0043] The preparation method of an indole iodide-pyridine bis-half-cyanine compound in this embodiment includes the following steps:

[0044] 3,5-pyridinedicarboxaldehyde (175.2 mg, 1.3 mmol) and 1,2,3,3-tetramethyl-3H-indole iodide (961.7 mg, 3.2 mmol) were weighed into a two-necked round-bottom flask, and 60 mL of ethanol was added. The mixture was reacted at 100 °C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain the crude product. The crude product was washed five times with acetonitrile and dried under vacuum to obtain 783.0 mg of the orange solid DII-3,5-Py, as shown in structural formula (II), with a yield of 85%.

[0045]

[0046] The compound shown in formula (Ⅰ) was subjected to HR-ESI-MS spectra, 1 The specific data from the 1H NMR spectroscopy test are shown below:

[0047] HR-ESI-MS m / z:calcd for C 31 H 33 I₂N₃, 478.2864[M⁻²I⁺CH₃O₂] - ] + ;found 478.3482

[0048] [M-2I+CH3O - ] + The specific spectrum is as follows Figure 4 As shown.

[0049] 1 H-NMR (400MH) Z DMSO-d6) δ: 9.48 (d, J = 1.6 Hz, 2H), 9.26 (s, 1H), 8.50 (d, J = 16.4 Hz, 2H), 8.01–7.93 (m, 6H), 7.71–7.69 (m, 4H), 4.26 (s, 6H), 1.85 (s, 12H). Specific spectra are shown below. Figure 5 As shown.

[0050] Example 3

[0051] The preparation method of an indole iodide-pyridine bis-half-cyanine compound in this embodiment includes the following steps:

[0052] Into a two-necked round bottom flask, 2,6-pyridine dicarboxaldehyde (134.1 mg, 1 mmol) and 1,2,3,3-tetramethyl-3H-indolium iodide (758.5 mg, 2.5 mmol) were weighed, 60 mL of ethanol was added, and the reaction was carried out at 70 °C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a crude product. The crude product was separated and purified by column chromatography (dichloromethane-methanol, volume ratio 20:1) to obtain a brick red solid DII-2,6-Py, which is a compound as shown in structural formula (I).

[0053]

[0054] Example 4

[0055] The preparation method of the indolium iodide-pyridine bis-hemicyanine compound in this example is as follows:

[0056] Into a two-necked round bottom flask, 2,6-pyridine dicarboxaldehyde (134.1 mg, 1 mmol) and 1,2,3,3-tetramethyl-3H-indolium iodide (758.5 mg, 2.5 mmol) were weighed, 60 mL of ethanol was added, and the reaction was carried out at 70 °C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a crude product. The crude product was separated and purified by column chromatography (dichloromethane-methanol, volume ratio 20:1) to obtain a brick red solid DII-2,6-Py, which is a compound as shown in structural formula (I).

[0057]

[0058] Example 5

[0059] The preparation method of the indolium iodide-pyridine bis-hemicyanine compound in this example is as follows:

[0060] Into a two-necked round bottom flask, 2,6-pyridine dicarboxaldehyde (134.1 mg, 1 mmol) and 1,2,3,3-tetramethyl-3H-indolium iodide (758.5 mg, 2.5 mmol) were weighed, 60 mL of ethanol was added, and the reaction was carried out at 70 °C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a crude product. The crude product was separated and purified by column chromatography (dichloromethane-methanol, volume ratio 20:1) to obtain a brick red solid DII-2,6-Py, which is a compound as shown in structural formula (I).

[0061]

[0062] Example 6

[0063] The preparation method of the indolium iodide-pyridine bis-hemicyanine compound in this example is as follows:

[0064] Into a two-necked round bottom flask, 3,5-pyridine dicarboxaldehyde (175.2 mg, 1.3 mmol) and 1,2,3,3-tetramethyl-3H-indolium iodide (961.7 mg, 3.2 mmol) were weighed, 60 mL of ethanol was added, and the reaction was carried out at 80°C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a crude product. The crude product was washed with acetonitrile five times, and dried under vacuum to obtain an orange solid DII-3,5-Py, which is a compound as shown in structural formula (II).

[0065]

[0066] Example 7

[0067] A method for preparing an indolium iodide-pyridine bis-hemicyanine compound in this example is as follows:

[0068] Into a two-necked round bottom flask, 3,5-pyridine dicarboxaldehyde (175.2 mg, 1.3 mmol) and 1,2,3,3-tetramethyl-3H-indolium iodide (961.7 mg, 3.2 mmol) were weighed, 60 mL of ethanol was added, and the reaction was carried out at 80°C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a crude product. The crude product was washed with acetonitrile five times, and dried under vacuum to obtain an orange solid DII-3,5-Py, which is a compound as shown in structural formula (II).

[0069]

[0070] Example 8

[0071] A method for preparing an indolium iodide-pyridine bis-hemicyanine compound in this example is as follows:

[0072] Into a two-necked round bottom flask, 3,5-pyridine dicarboxaldehyde (175.2 mg, 1.3 mmol) and 1,2,3,3-tetramethyl-3H-indolium iodide (961.7 mg, 3.2 mmol) were weighed, 60 mL of ethanol was added, and the reaction was carried out at 80°C for 12 h under argon protection. The reaction solution was cooled to room temperature, and the solvent was removed by rotary evaporation to obtain a crude product. The crude product was washed with acetonitrile five times, and dried under vacuum to obtain an orange solid DII-3,5-Py, which is a compound as shown in structural formula (II).

[0073]

[0074] Test Example

[0075] (1) Inhibition of the proliferation of breast cancer, lung cancer, colorectal cancer and other cells by DII-2,6-Py, DII-3,5-Py and 5Fu

[0076] The breast cancer, lung cancer and colorectal cancer cell proliferation inhibition effects of DII-2,6-Py prepared in Example 1, DII-3,5-Py prepared in Example 2 and 5Fu were tested, and the specific testing process is shown as follows:

[0077] The MDA-MB-31, BT549, SW480, DLD-1, A549 and H460 cells in logarithmic growth phase were collected respectively, counted, and the cell suspension concentration was adjusted to 5x10 4 The compound DII-2,6-Py, DII-3,5-Py and 5Fu were diluted with DMSO and added to the culture hole, so that the final concentration of the compound in the system was 0.195, 0.39, 0.78, 1.56, 3.12, 6.25, 12.5, 25, 50 and 100 (μmol / L) respectively. After 48h of continuous culture, 10μL of MTT solvent (concentration of 5mg / ml) was added to each hole, and incubated at 37℃ for 4h. The culture solution containing MTT was aspirated, and 100μL of DMSO was added to each hole. After low-speed shaking on a shaking table for 10min, the OD value at an absorption wavelength of 490nm was measured by an enzyme marker, and the results were recorded. The cell growth curve was drawn with the dose of the compound as the horizontal coordinate and the absorbance value as the vertical coordinate. The statistical results of the half-inhibition rate (IC50 value) of the compound on tumor cells are shown in Table 1. Figures 6-8 and Table 1:

[0078] The statistical results of the half-inhibition rate (IC50 value) of the compound on tumor cells are shown in Table 1.

[0079]

[0080] As Figures 6-8 and Table 1: the IC50 of 5Fu in MDA-MB-31, BT549, SW480 and A549 cell lines is about 17.9 times, 6.57 times, 1.79 times, 38.57 times and 4.5 times that of DII-2,6-Py, and about 26.04 times, 11.43 times, 3.44 times, 20.08 times and 6.29 times that of DII-3,5-Py. Therefore, in this experiment, in addition to the colon cancer DLD-1 cells, the sensitivity of DII-2,6-Py and DII-3,5-Py to the remaining tumor cells (especially breast cancer and lung cancer cells) is obviously better than that of 5Fu, which can significantly inhibit the proliferation of these cells.

[0081] (2) Mitochondrial co-localization experiment of DII-2,6-Py and DII-3,5-Py

[0082] Mitochondrial co-localization experiment of DII-2,6-Py prepared in Example 1 and DII-3,5-Py prepared in Example 2 was carried out as follows: MDA-MB-231 cells were seeded in 35 mm culture dishes (2 x 104cell / dish), and after 24 h incubation in an incubator, the old medium was replaced with fresh medium. DII-2,6-Py (5 μM), DII-3,5-Py (5 μM) and DMSO were added respectively, and incubated for 30 min. After the fresh medium was replaced, the mitochondria dye Mito-Tracker Red (0.5 μM) was added to incubate the cells for 30 min. After the cells were washed with PBS, fluorescence imaging was carried out by laser confocal (FV1000, Olympus, Japan), and the results are shown in Figure 9

[0083] The results show that after DII-2,6-Py and DII-3,5-Py treatment, the green fluorescent part of the compounds in the cells is consistent with the part positively stained by Mito-Tracker Red. And the state of the cells is better, which shows the sensitivity of the compounds to the mitochondrial localization of tumor cells at low doses.

[0084] The above results show that DII-2,6-Py and DII-3,5-Py can significantly inhibit the proliferation of breast cancer, lung cancer and colorectal cancer cells, and the drug efficacy is significantly better than that of the clinical drug 5Fu, and has mitochondrial targeting. Therefore, such drugs have good anticancer effect and development potential.

[0085] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.​

Claims

1. An indole iodide-pyridine bishemine compound, characterized in that, It has a structural formula as shown in formula (I) or formula (II):

2. The method for preparing the indole iodide-pyridine bishemine compound according to claim 1, characterized in that, The steps are as follows: 1,2,3,3-tetramethyl-3H-indole iodide and pyridinedicarboxaldehyde are dissolved in a solvent and reacted under argon protection. After the reaction is completed, indole iodide-pyridinedihalocyanine compound is obtained.

3. The method for preparing the indole iodide-pyridine bishemine compound according to claim 2, characterized in that, The pyridinedicarboxaldehyde is 2,6-pyridinedicarboxaldehyde or 3,5-pyridinedicarboxaldehyde.

4. The method for preparing the indole iodide-pyridine bishemine compound according to claim 3, characterized in that, The solvent is ethanol, methanol, or acetone.

5. The method for preparing the indole iodide-pyridine bishemine compound according to claim 4, characterized in that, The molar ratio of 1,2,3,3-tetramethyl-3H-indole iodide to pyridinedicarboxaldehyde is (2.2-2.5):

1.

6. The method for preparing the indole iodide-pyridine bishemine compound according to claim 5, characterized in that, The concentration of pyridinedicarboxaldehyde is 0.017-0.024 mol / L.

7. The method for preparing the indole iodide-pyridine bishemine compound according to any one of claims 2-6, characterized in that, The reaction temperature is 70-100℃, and the reaction time is 6-18h.

8. The method for preparing the indole iodide-pyridine bishemine compound according to claim 7, characterized in that, It also includes post-processing steps after the reaction: when the reactant is 2,6-pyridinedicarboxaldehyde, the solvent is removed by rotary evaporation and purified by column chromatography; when the reactant is 3,5-pyridinedicarboxaldehyde, the solvent is removed by rotary evaporation, and the reactants are washed with acetonitrile and dried.

9. The use of the indole iodide-pyridine bishemine compound of claim 1, and its biologically acceptable salts and formulations, in the preparation of cancer therapeutic drugs.

10. The application according to claim 9, characterized in that, The cancers referred to are breast cancer, lung cancer, and colorectal cancer.

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