Synthesis of porphyrin derivatives and their anti-tumor applications

Abstract

The application belongs to the field of medicine, and particularly relates to a preparation method and anti-tumor application of a novel hirsutellin derivative. The novel hirsutellin derivative with a novel structure is prepared by using a simple and efficient synthesis method, and the specific structure is as follows: the in-vitro anti-tumor activity research of the compound in the application shows that the novel hirsutellin derivative has obvious inhibitory effect on the growth of human breast cancer cells (MDA-MB-231) and human gastric cancer cells (SGC7901), which shows that the hirsutellin derivative can be used as a lead compound or a candidate compound for development of an anti-tumor drug.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of medicine, and particularly relates to synthesis and anti-tumor application of a novel calobrin derivative. BACKGROUND

[0002] At present, chemotherapy drugs include alkylating agents (cyclophosphamide) and antibiotics (doxorubicin). Since these chemotherapy drugs have no differential effects on tumors and normal cells, side effects such as neurotoxicity exist. Therefore, it is necessary to develop new anti-tumor drugs with better efficacy and less toxicity. Calobrin, as one of natural products derived from Dendrobium huoshanense, has significant pharmacological effects such as anti-cancer, antibacterial and antioxidant effects. In recent years, calobrin has been proved to have significant anti-tumor effects on various types of cancers in vivo and in vitro through various mechanisms, and is considered as a potential anticancer molecule. However, calobrin has the disadvantages of low solubility, fast metabolism and poor bioavailability, which makes it unable to become an anti-tumor drug with high efficiency and low toxicity. Therefore, it is of great research value to develop new calobrin derivatives based on calobrin and study the anti-tumor activity thereof, which is beneficial to the development of self-owned proprietary drugs in China. SUMMARY

[0003] The application aims to provide a calobrin derivative with a novel structure and a simple preparation method, and another purpose of the application is to provide the application of the calobrin derivative in anti-tumor drugs.

[0004] To achieve the above-mentioned purposes, the calobrin derivative has the following general structure:

[0005]

[0006] Specifically selected from the following compounds:

[0007]

[0008] The application also provides a preparation method of the calobrin derivative, which comprises the following steps: first, a substitution reaction of benzyl bromide WCJ-05 containing a substituent group and aniline WCJ-01 containing a substituent group in acetone to prepare a compound WCJ-02; a reaction of the compound WCJ-02 with 4-chloroformyl benzoic acid methyl ester or 4-bromomethyl benzoic acid methyl ester or succinic acid methyl ester acyl chloride or oxalyl chloride monomethyl ester to prepare a compound WCJ-03; a hydrolysis reaction of the compound WCJ-03 with sodium hydroxide aqueous solution to prepare a compound WCJ-04; and a condensation reaction of the compound WCJ-04 with 4-fluorobenzhydrazide or benzhydrazide to prepare a target product calobrin derivative.

[0009] The synthesis route is as follows:

[0010] The application aims to provide a calobrin derivative with a novel structure and a simple preparation method, and another purpose of the application is to provide the application of the calobrin derivative in anti-tumor drugs.

[0011] This invention also provides the use of any of the above-mentioned compounds in the preparation of tumor drugs. Preferably, the tumor is selected from human breast cancer cells (MDA-MB-231) and human gastric cancer cells (SGC7901).

[0012] The present invention also provides an antitumor drug comprising any of the above-mentioned dendritic derivatives and a pharmaceutically acceptable carrier, pharmaceutical salt, and excipient.

[0013] The present invention has the following beneficial effects:

[0014] This invention provides a novel dendritic derivative.

[0015] This invention provides a method for synthesizing a dendritic derivative. The method is simple to operate, safe, produces few reaction byproducts, has a high yield, and the product is easy to separate and purify.

[0016] In vitro antitumor activity experiments of the novel romaine derivatives of this invention revealed that these compounds exhibit strong inhibitory activity against human gastric cancer cells (SGC-7901), with the strongest inhibitory activity reaching IC50. 50 With a value of 23 nM, it has broad application prospects in the preparation of anti-tumor drugs. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly described below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 For compound (WCJ-02-3) 1 H-NMR

[0019] Figure 2 For compound (WCJ-02-3) 13 C-NMR

[0020] Figure 3 HRMS of compound (WCJ-02-3) Detailed Implementation

[0021] The technical solutions of the present invention will be clearly and completely described below in conjunction with specific embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention or its application or use. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0022] The structures of the compounds of this invention were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR measurements were performed using a Bruker AV-500 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD) as the solvents, and tetramethylsilane (TMS) as the internal standard. Chemical shifts (δ) are expressed in ppm.

[0023] Examples of compound synthesis

[0024] Example 1: Preparation of N-(3,4-dimethoxybenzyl)-4-(2-(4-fluorobenzoyl)hydrazine-1-carbonyl)-N-(3,4,5-trimethoxyphenyl)benzamide (WCJ-02-17)

[0025] Preparation of 3,4,5-trimethoxy-N-(3,4-dimethoxybenzyl)aniline (WCJ-02)

[0026] In a 100 mL round-bottom flask, 3,4,5-trimethoxyaniline (5.38 g, 29.37 mmol), 3,4-dimethoxybenzyl bromide (6.79 g, 29.37 mmol), potassium carbonate (4.46 g, 32.30 mmol), and 60 mL of acetone were added and the mixture was heated under reflux for 8 h. The reaction was stopped by TLC monitoring until the reaction was complete. The mixture was extracted with dichloromethane and saturated brine, and the organic phase was retained. The organic phase was dried over anhydrous sodium sulfate, and the mixture was separated by vacuum distillation and silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 15:1) to obtain intermediate WCJ-02.

[0027] Preparation of methyl 4-((3,4-dimethoxybenzyl)(3,4,5-trimethoxyphenyl)carbamoyl)benzoate (WCJ-03)

[0028] 50 mL of dichloromethane and 5.74 mL of N,N-diisopropylethylamine (32.96 mmol) were added to a 100 mL round-bottom flask containing intermediate WCJ-02 (5 g, 15.00 mmol). The flask was cooled to 0 °C in an ice-water bath, and 10 mL of dichloromethane containing methyl 4-chloroformylbenzoate (3.6 g, 18.00 mmol) was slowly added dropwise while stirring. The reaction was stopped after TLC monitoring until the reaction was complete. The mixture was extracted with dichloromethane and saturated brine, and the organic phase was retained. The organic phase was dried over anhydrous sodium sulfate, distilled under reduced pressure, and separated by silica gel column chromatography (eluent: dichloromethane:methanol = 40:1) to obtain intermediate WCJ-03 (6.2 g, 79%).

[0029] Preparation of 4-((3,4-dimethoxybenzyl)(3,4,5-trimethoxyphenyl)carbamoyl)benzoic acid (WCJ-04)

[0030] 20 mL of methanol and 5 mL of water were added to a 50 mL round-bottom flask containing intermediate WCJ-03 (4 g, 8.59 mmol) and stirred at room temperature to dissolve. 3 mL of an aqueous solution of sodium hydroxide (378 mg, 9.45 mmol) was added and stirred at room temperature. The reaction was stopped after TLC monitoring showed that the reaction was complete. The mixture was then distilled under reduced pressure, extracted with ethyl acetate and water, and the aqueous phase was retained. The pH was adjusted to 1 with 1 M hydrochloric acid, and the organic phase was extracted with dichloromethane and dried over anhydrous sodium sulfate. The mixture was then distilled under reduced pressure to obtain intermediate WCJ-04 (3.4 g, 88%).

[0031] Preparation of N-(3,4-dimethoxybenzyl)-4-(2-(4-fluorobenzoyl)hydrazine-1-carbonyl)-N-(3,4,5-trimethoxyphenyl)benzamide (WCJ-02-17)

[0032] Intermediate WCJ-04 (250 mg, 553.74 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (143.83 mg, 609.11 μmol), and 1-hydroxybenzotriazole (68.32 mg, 609.11 μmol) were added to a 25 mL round-bottom flask containing 10 mL of dichloromethane and stirred at room temperature for 30 minutes. 4-fluorobenzoylhydrazine (151 mg, 0.98 mmol) was added, and the reaction was stopped by TLC monitoring. The mixture was then extracted with dichloromethane and water, and the organic phase was retained. The extract was dried over anhydrous sodium sulfate, and the mixture was separated by vacuum distillation and silica gel column chromatography (eluent: dichloromethane:methanol = 40:1) to obtain the white solid target product WCJ-02-17.

[0033] White solid, yield: 67.7%. mp 98.5-99.5℃. 1 H NMR (400MH) z, CDCl3) δ9.34 (d, J=4.6Hz, 1H), 9.22 (s, 1H), 7.88-7.84 (m, 2H), 7.68 (d, J=8.3Hz, 2H), 7.44 (d, J=8.4Hz, 2H), 7.16-7.0 9(m, 2H), 6.90(s, 1H), 6.85-6.78(m, 2H), 6.08(s, 2H), 5.00(s, 2H), 3.87(s, 3H), 3.83(s, 3H), 3.77(s, 3H), 3.58(s, 6H). 13 C NMR (101MHz, CDCl3) δ169.26, 166.48, 164.28, 164.16, 163.96, 153.23, 148.94, 148.61, 140.17, 138.23, 137.16, 131.97, 129.93, 129.8 4, 128.46, 127.36, 127.33, 126.95, 121.37, 115.90, 115.68, 112.16, 110.95, 105.66, 60.97, 56.16, 55.92, 53.77. HR-MS (ESI) m / z: calcd for C 33 H 33 N3O9Na[M+Na] + 634.2205, found 634.2201.

[0034] Example 2: Preparation of N-(3-fluoro-4-methoxybenzyl)-4-(2-(4-fluorobenzoyl)hydrazide-1-carbonyl)-N-(3,4,5-trimethoxyphenyl)benzamide (WCJ-02-21)

[0035] The preparation method is the same as in Example 1, using 3-fluoro-4-methoxybenzyl bromide instead of p-3,4-dimethoxybenzyl bromide.

[0036] White solid, yield: 75%. mp 100.2-101.2℃. 1 H NMR (400MHz, CDCl3) δ9.36 (s, 1H), 9.27 (s, 1H), 7.89-7.82 (m, 2H), 7.67 (d, J=8.1Hz, 2H), 7.44 (d, J=8.0Hz, 2H), 7.12 (t, J= 8.6Hz, 3H), 7.01 (d, J=8.3Hz, 1H), 6.89 (t, J=8.5Hz, 1H), 6.08 (s, 2H), 4.98 (s, 2H), 3.88 (s, 3H), 3.77 (s, 3H), 3.60 (s, 6H). 13C NMR (101MHz, CDCl3) δ169.31, 166.49, 164.25, 164.16, 163.97, 153.39, 15 3.33, 150.94, 147.24, 147.13, 139.89, 138.03, 137.24, 132.07, 130.15, 12 9.93, 129.84, 128.52, 127.35, 126.96, 124.82, 116.69, 116.51, 115.91, 1 15.69, 113.20, 105.53, 60.98, 56.27, 56.18, 53.23. HR-MS (ESI) m / z: calcd for C 32 H 29 N3O7F2Na[M+Na] + 628.1871, found 628.1871.

[0037] Example 3: Preparation of 4-(2-(4-fluorobenzoyl)hydrazide-1-carbonyl)-N-(4-methoxy-3-nitrobenzyl)-N-(3,4,5-trimethoxyphenyl)benzamide (WCJ-02-19)

[0038] The preparation method is the same as in Example 1, using 4-methoxy-3-nitrobenzyl bromide instead of p-3,4-dimethoxybenzyl bromide.

[0039] White solid, yield: 72.8%. mp 120.8-121.8℃. 1 H NMR (400MHz, CDCl3) δ9.31 (d, J=5.3Hz, 1H), 9.28 (s, 1H), 7.85 (ddd, J=10.0, 5.2, 2.6Hz, 2H), 7.79 (d, J=2.3Hz, 1H), 7.68 (d, J=8.4Hz, 2H), 7.58 (dd, J= 8.3, 2.1Hz, 1H), 7.44 (d, J=8.4Hz, 2H), 7.13 (t, J=8.6Hz, 2H), 7.05 (d, J=8. 7Hz, 1H), 6.11(s, 2H), 5.04(s, 2H), 3.96(s, 3H), 3.78(s, 3H), 3.63(s, 6H). 13C NMR (101MHz, CDCl3) δ169.34, 164.01, 163.93, 153.57, 152.40, 139.51, 139.37, 137.86, 137.46, 134.86, 132.24, 129.88, 129.7 9, 129.56, 128.63, 127.35, 126.97, 125.95, 116.00, 115.78, 113.65, 105.48, 61.02, 56.61, 56.25, 52.94. HR-MS (ESI) m / z: calcd forC 32 H 29 N4O9FNa[M+Na] + 655.1816, found 655.1816.

[0040] Example 4: Preparation of 4-(2-(4-fluorobenzoyl)hydrazide-1-carbonyl)-N-(4-methoxybenzyl)-N-(3,4,5-trimethoxyphenyl)benzamide (WCJ-02-3)

[0041] The same preparation method as in Example 1 was used, replacing p-3,4-dimethoxybenzyl bromide with 4-methoxybenzyl bromide.

[0042] White solid, yield: 72%. mp 108.5-109.5℃. 1 H NMR (500MHz, CDCl3) δ9.52 (s, 1H), 9.28 (s, 1H), 7.77 (d, J = 8.2Hz, 2H), 7.65 (d, J = 7.9Hz, 2H), 7.41 (dd, J = 11.2, 8.3Hz , 4H), 7.24 (d, J=7.9Hz, 2H), 6.84 (d, J=8.2Hz, 2H), 6.06 (s, 2H), 5.00 (s, 2H), 3.79 (s, 3H), 3.76 (s, 3H), 3.57 (s, 6H). 13 C NMR (101MHz, CDCl3) δ169.31, 166.47, 164.40, 164.26, 163.94, 159.14, 153.21, 140.16, 138.13, 137.12, 131.94, 130.20, 129.96, 12 9.87, 129.39, 128.45, 127.41, 127.37, 126.94, 115.87, 115.65, 113.89, 105.68, 60.96, 56.14, 55.28, 53.43. HR-MS (ESI) m / z: calcd for C 32 H 30N3O7NaF[M+Na] + 610.1969, found 610.1965.

[0043] Example 5: Preparation of 4-(2-(4-fluorobenzoyl)hydrazide-1-carbonyl)-N-(4-methoxyphenyl)-N-(3,4,5-trimethoxybenzyl)benzamide (WCJ-02-18)

[0044] The preparation method is the same as in Example 1, using 3,4,5-trimethoxybenzyl bromide instead of p-3,4-dimethoxybenzyl bromide and 4-methoxyaniline instead of 3,4,5-trimethoxyaniline.

[0045] White solid, yield: 56.9%. mp 107.5-108.5℃. 1 H NMR (400MHz, CDCl3) δ9.48 (s, 1H), 9.22 (s, 1H), 7.87-7.82 (m, 2H), 7.63 (d, J = 8.0Hz, 2H), 7.37 (d, J = 8.0Hz, 2H), 7.10 (t, J=8.6Hz, 2H), 6.79 (d, J=8.4Hz, 2H), 6.69-6.65 (m, 2H), 6.50 (s, 2H), 4.98 (s, 2H), 3.84 (s, 3H), 3.78 (s, 6H), 3.72 (s, 3H). 13 C NMR (101MHz, CDCl3) δ169.48, 166.44, 164.59, 164.28, 163.92, 158.44, 153.17, 140.09, 137.42, 135.27, 132.76, 131.75, 129.96, 12 9.87, 129.18, 128.74, 127.32, 127.29, 126.90, 115.82, 115.61, 114.40, 105.93, 60.88, 56.12, 55.32, 54.17. HR-MS (ESI) m / z: calcd for C 32 H 30 N3O7FNa[M+Na] + 610.1965, found 610.1965.

[0046] Example 6: Preparation of 2-(2-(4-fluorobenzoyl)hydrazino)-N-(4-methoxybenzyl)-2-oxo-N-(3,4,5-trimethoxyphenyl)acetamide (WCJ-02-22)

[0047] The same preparation method as in Example 1 was used, replacing p-3,4-dimethoxybenzyl bromide with 4-methoxybenzyl bromide and methyl 4-chloroformylbenzoate with methyl oxaloyl chloride monomethyl ester.

[0048] White solid, yield: 42.4%. mp 98.5-99.5℃. 1 H MR (400MHz, CDCl3) δ9.57 (d, J=5.4Hz, 1H), 8.58 (d, J=5.2Hz, 1H), 7.74 (dd, J=8.8, 5.2Hz, 2H), 7.16 (d, J=8.6Hz, 2 H), 7.08 (t, J=8.6Hz, 2H), 6.82 (d, J=8.6Hz, 2H), 6.20 (s, 2H), 4.85 (s, 2H), 3.82 (s, 3H), 3.79 (s, 3H), 3.71 (s, 6H). 13 C NMR (101MHz, CDCl3) δ165.76, 165.46, 163.78, 161.65, 159.43, 157.59, 153.27, 137.71, 135.38, 130.57, 130.22, 13 0.02, 129.95, 129.58, 129.28, 116.00, 115.82, 113.86, 104.47, 60.96, 56.13, 55.29, 54.89. HR-MS (ESI) m / z: calcd forC 26 H 26 N3O7FNa[M+Na] + 534.1657, found 534.1653

[0049] Example 7: Preparation of 4-(2-(4-fluorobenzoyl)hydrazino)-N-(4-methoxybenzyl)-4-oxo-N-(3,4,5-trimethoxyphenyl)butyramide (WCJ-02-23)

[0050] The preparation method is the same as in Example 1, using 4-methoxybenzyl bromide instead of 3,4-dimethoxybenzyl bromide and methyl succinate chloride instead of methyl succinate.

[0051] White solid, yield: 18.3%. mp 84.5-85.5℃. 1H NMR (400MHz, CDCl3) δ8.78 (s, 1H), 7.87-7.82 (m, 2H), 7.15-7.07 (m, 5H), 6.79-6.77 (m, 2H), 6.20 (s , 2H), 4.78 (s, 2H), 3.84 (s, 3H), 3.75 (s, 3H), 3.71 (s, 6H), 2.67-2.63 (m, 2H), 2.53 (t, J=5.9Hz, 2H). 13 C NMR (101MHz, CDCl3) δ172.00, 171.26, 166.34, 164.24, 163.82, 159.03, 153.61, 137.81, 137.14, 130.32, 129.89, 129.80, 1 29.46, 128.00, 127.97, 115.80, 115.58, 113.71, 105.75, 60.95, 56.19, 55.24, 52.62, 29.53, 29.32. HR-MS (ESI) m / z: calcd for C 28 H 30 N3O7FNa[M+Na] + 562.1970, found 562.1965.

[0052] Example 8: Preparation of 4-fluoro-N'-(4-((4-methoxybenzyl)(3,4,5-trimethoxyphenyl)amino)methyl)benzoyl)benzoylhydrazine (DX-30-01)

[0053] The preparation method is the same as in Example 1, using 4-methoxybenzyl bromide instead of 3,4-dimethoxybenzyl bromide and 4-bromomethylbenzoate instead of methyl 4-chloroformylbenzoate.

[0054] White solid, yield: 80.9%. mp 136.5-137.5℃. 1 H NMR (400MHz, CDCl3) δ9.70 (s, 1H), 9.44 (s, 1H), 7.89-7.85 (m, 2H), 7.81 (d, J=8.3Hz, 2H), 7.33 (d, J=7.8Hz, 2H), 7.26 (s, 2H), 7.17 (d, J=8. 8Hz, 2H), 7.08 (td, J=8.6, 1.6Hz, 2H), 6.87 (d, J=8.8Hz, 2H), 5.94 (s, 2H), 4.58 (s, 2H), 4.52 (s, 2H), 3.80 (s, 3H), 3.75 (s, 3H), 3.68 (s, 6H). 13C NMR (101MHz, CDCl3) δ166.48, 164.99, 164.04, 163.96, 158.81, 153.74, 145.93, 144.20, 130.28, 130.18, 129.94, 129.90, 129. 81, 128.15, 127.75, 127.37, 127.12, 115.89, 115.67, 114.12, 91.53, 61.06, 55.97, 55.30, 54.80, 54.62. HR-MS (ESI) m / z: calcd for C 32 H 33 N3O6F[M+H] + 574.2353, found 574.2353.

[0055] Example 9: Preparation of N'-benzoyl-4-(((4-methoxybenzyl)(3,4,5-trimethoxyphenyl)amino)methyl)benzoylhydrazine (DX-33-01)

[0056] The preparation method is the same as in Example 1, using 4-methoxybenzyl bromide instead of p-3,4-dimethoxybenzyl bromide, methyl 4-bromomethylbenzoate instead of methyl 4-chloroformylbenzoate, and benzoyl hydrazine instead of 4-fluorobenzoyl hydrazine.

[0057] White solid, yield: 37.4%. mp 115-116℃. 1 H NMR (400MHz, CDCl3) δ9.41 (q, J=6.3Hz, 2H), 7.88-7.82 (m, 4H), 7.58-7.53 (m, 1H), 7.45 (t, J=7.6Hz, 2H), 735 (d, J=8.0Hz, 2 H), 7.18 (d, J=8.6Hz, 2H), 6.89-6.85 (m, 2H), 5.95 (s, 2H), 4.59 (s, 2H), 4.53 (s, 2H), 3.80 (s, 3H), 3.76 (s, 3H), 3.68 (s, 6H). 13 C NMR (101MHz, CDCl3) δ164.40, 164.04, 158.81, 153.74, 132.49, 131.28, 130.07, 128.78, 128.17, 1 27.70, 127.28, 127.18, 114.12, 91.51, 61.07, 55.98, 55.32, 54.80, 54.70. HR-MS (ESI) m / z: calcd for C 32 H 34 N3O6[M+H] +556.2448, found 556.248.

[0058] Application examples of compounds

[0059] In vitro antitumor activity test: Four cell lines were used in the CCK8 assay, namely human breast cancer cells (MDA-MB-231) and human gastric cancer cells (SGC-7901).

[0060] Collect cells in the logarithmic growth phase, adjust the cell suspension concentration, add 100 μL to each well, and seed the cells to adjust the cell density. Fill the edge wells of the 96-well plate with PBS. Incubate for 24 h at 5% CO2, 37°C, and 90% humidity until the cell monolayer covers the bottom of the 96-well plate. Add a concentration gradient of the synthesized drug of this invention, with 9 concentrations, 200 μL in each well (3 replicates), and incubate for 72 h. Observe under an inverted microscope. Add 10 μL of CCK8 solution to each well and continue culturing for 2 h. Measure the absorbance of each well at 450 nm using a microplate reader. Statistically analyze the experimental results and calculate the IC50 using GraphPad Prism 9.5 software. 50 The (μmol / L) values ​​are shown in Table 1 below.

[0061] Table 1 Results of in vitro antitumor activity tests

[0062]

Claims

1. A novel dendritic derivative, characterized in that, It has the structure described in general formula I: Specifically selected from the following compounds:

2. The application of the dendritic derivative as described in claim 1 in drug preparation, characterized in that, It is used as an active ingredient in the preparation of drugs for treating breast cancer cells and gastric cancer.

3. An antitumor drug composition, characterized in that, The pharmaceutical composition uses the compound of claim 1 as the active ingredient; the pharmaceutical composition further includes a pharmaceutically acceptable carrier, pharmaceutical salt, or excipient.

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