Porphyrin-formononetin derivative and anti-tumor application thereof

Abstract

The porphyrin is bonded with the natural product isoflavone formononetin with antitumor activity, so that on one hand, the PDT effect of the porphyrin can be played to inhibit the growth and reproduction of tumor cells, and on the other hand, the tumor affinity characteristic of the porphyrin can be played, the formononetin with antitumor activity is positioned to a tumor part, the damage to normal tissues is reduced, and the antitumor activity of the formononetin is improved. The cytotoxicity is reduced.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and particularly to a porphyrin-formononetin derivative and its anti-tumor use. Background Art

[0002] Photodynamic Therapy (PDT), as an emerging reactive oxygen species (ROS)-mediated non-invasive therapy, has received increasing attention in recent years. It has now been applied to the treatment of cancers such as laryngeal cancer, skin cancer, breast cancer, etc., and certain therapeutic effects have been achieved. Since PDT can selectively act on tumor tissues and hardly affect the functions of normal tissue cells, it has the advantage of low toxicity and side effects, and is particularly suitable for people intolerant to traditional cancer treatment methods such as chemotherapy and radiotherapy. The reactive oxygen species generated during the PDT process (including hydroxyl radicals generated by type I and singlet oxygen generated by type II) can oxidize biological macromolecules (proteins, nucleic acids, lipids) in the system. After these essential biological macromolecules are oxidized, the normal life activities of cells are interfered, thereby causing apoptosis and necrosis of tumor cells. To fully exert the therapeutic efficacy of PDT, a suitable photosensitizer must be selected first.

[0003] With the development of PDT, remarkable progress has been made in the preparation of photosensitizers, especially porphyrin-based photosensitizers. Most photosensitizers used for cancer treatment have a porphyrin-based macrocyclic skeleton. The main advantages of porphyrin compounds in photodynamic research include: 1) the stability of aromatic compounds; 2) effective absorption of visible light; 3) high reactive oxygen species yield; 4) easy functionalization modification and structural diversity; 5) relatively long triplet lifetime and low dark toxicity. The first photosensitive drug Photofri approved by the US FDA for marketing is a classic porphyrin-structured drug.

[0004] Li et al. (Chinese Chemical Letters, 2007, 18(11): 1331-1334) designed and synthesized a novel potential targeted anticancer drug based on porphyrin and 5-fluorouracil, and evaluated the in vitro anticancer activity against human hepatoma cell line SMCC-7721 by MTT method. The preliminary results showed that the anticancer activities of the conjugates were more than twice that of 5-fluorouracil. Porphyrin had no killing effect on tumor cells under dark conditions, indicating that the conjugates actually improved the targeting of 5-fluorouracil to tumor tissues, thus enhancing the anticancer activity. S. Weimin et al. (Bioorg Med Chem, 2008, 16(10): 5665-5671) coupled 5-fluoroacetic acid / Boc-L-phenylalanine with amino porphyrin to synthesize a series of porphyrin compounds containing 5-fluorouracil / L-phenylalanine. The results of in vitro anticancer activity studies showed that the introduction of 5-fluorouracil and L-phenylalanine could significantly improve the phototoxicity of porphyrin. The high selectivity of porphyrin compounds for tumor tissues broadened the prospects for the research of anticancer drug-linked porphyrin compounds.

[0005] Formononetin (FMN), namely 7-hydroxy-4'-methoxyisoflavone, also known as biochanin A and formononetin. FMN is a phytoestrogen that can exert estrogen-like effects due to its similar structure to mammalian estrogens and shows certain therapeutic effects on estrogen-related diseases. Some studies have shown that FMN has various pharmacological activities, multiple benefits and medical values, and has extensive applications in medicine. Such as anticancer, neuroprotection, antidiabetes, anti-atherosclerosis, anti-inflammatory, etc. With the discovery of the anti-tumor activity of FMN, this property has become a research hotspot for formononetin.

[0006] Some studies have shown that FMN can be used for the prevention and treatment of various cancers and has shown potential application prospects in the field of anticancer, such as cervical cancer, lung cancer, bladder cancer, prostate cancer, breast cancer, colon cancer, gastric cancer, nasopharyngeal cancer, etc. FMN inhibits the growth and proliferation of cancer cells through multiple mechanisms, induces cell cycle arrest, apoptosis, and inhibits the migration, invasion and angiogenesis of cancer cells by regulating signal pathways. In addition, the antioxidant and anti-inflammatory effects of FMN are of great significance for the prevention and treatment of cancer. The occurrence of cancer is partly due to cell damage and mutations caused by oxidative stress and inflammatory responses in the body, and the antioxidant and anti-inflammatory properties of FMN help reduce this damage and lower the risk of cancer occurrence. Studies have found that FMN can enhance the immune system function and improve the body's anti-cancer ability. The immune system plays a crucial role in fighting cancer, and the immunomodulatory effect of FMN can help enhance the body's resistance and strengthen the immune system's ability to recognize and eliminate cancer cells.

[0007] In the present invention, porphyrin is linked with formononetin, a natural product with anti-tumor activity. On the one hand, the PDT effect of porphyrin can be exerted to inhibit the growth and reproduction of tumor cells, and at the same time, its tumor-affinity characteristic can be utilized to localize formononetin with anti-tumor activity to the tumor site, reducing the damage to normal tissues and lowering the cytotoxicity. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide a porphyrin-formononetin derivative, which can, on the one hand, exert the PDT effect of porphyrin to inhibit the growth and reproduction of tumor cells, and at the same time, utilize its tumor-affinity characteristic to localize formononetin with anti-tumor activity to the tumor site, reducing the damage to normal tissues and lowering the cytotoxicity.

[0009] In the first aspect of the present invention, there is provided a compound represented by Formula I or Formula II and a pharmaceutically acceptable salt thereof, which has the following structure:

[0010]

[0011] Wherein, n is an integer selected from 1 to 8;

[0012] Each R independently is selected from H, halogen, hydroxyl, nitro, CN, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 6-10 aryl, C 2-10 heteroaryl;

[0013] R' is selected from H, halogen, hydroxyl, nitro, CN, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl;

[0014] M is selected from Zn, Ni, Mn.

[0015] Preferably, n is selected from 1, 2, 3, 4 or 5.

[0016] Preferably, each R independently is selected from H, halogen, hydroxyl, nitro, CN, C 1-6 alkyl; more preferably, each R independently is selected from H, Br or Cl;

[0017] Preferably, R' is selected from H, halogen, hydroxyl, nitro, CN, C 1-6 alkyl; more preferably, R is selected from H;

[0018] Preferably, M is selected from Zn.

[0019] Another aspect of the present invention provides a method for preparing a compound of formula I, and its synthetic route is as follows:

[0020]

[0021] Wherein, the definitions of n, R, and R' are as described above.

[0022] The specific reaction steps are as follows:

[0023] In an organic solvent, add a base and porphyrin 1, heat and stir, and add the formononetin derivative 2 to the reaction system until the reaction is complete. After post-treatment, a porphyrin-formononetin derivative of formula I is obtained.

[0024] Preferably, the molar ratio of porphyrin derivative 1 to formononetin derivative 2 is: 1:(1 - 1.5), preferably 1:1 - 1.2, and more preferably 1:1.2

[0025] The base is selected from potassium hydroxide, triethylamine or potassium carbonate, and more preferably potassium carbonate.

[0026] Another aspect of the present invention provides a method for preparing a compound of formula II, and its synthetic route is as follows:

[0027]

[0028] Wherein, the definitions of n, R, R' and M are as described above.

[0029] The specific reaction steps are as follows:

[0030] Dissolve the compound of formula I in an organic solvent, add the M metal salt, reflux and react, monitor by TLC until the reaction is complete, and after post-treatment, a porphyrin-formononetin derivative of formula II is obtained.

[0031] Preferably, the molar ratio of the compound of formula I to the M metal salt is: 1:3 - 7, preferably 1:5.

[0032] Another aspect of the present invention provides a pharmaceutical composition, which comprises a compound of formula I or formula II or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient.

[0033] Another aspect of the present invention relates to the use of a compound of formula I or formula II or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same in the preparation of an anti-cancer drug;

[0034] Preferably, the cancer is selected from lung cancer, breast cancer, colon cancer, gastric cancer, prostate cancer, and bladder cancer.

[0035] Compared with the prior art, the beneficial effects of the present invention are:

[0036] (1) The present invention provides a new class of porphyrin-formononetin derivatives with anti-cancer activity, broadening the scope of existing anti-cancer compounds and can be further optimized as lead compounds.

[0037] (2) The compounds of the present invention use porphyrin molecules as carriers. Utilizing their effect of aggregating in tumor tissues, they have targeting properties towards tumor cells, reducing the side effect of killing normal cells.

[0038] (3) The porphyrin-formononetin derivatives of the present invention can achieve the synergistic effect of phototherapy and chemotherapy. In addition, inserting metal Zn into porphyrin can also enhance the anti-tumor activity of the compounds. Description of the Drawings

[0039] Figure 1 Trend chart of the fluorescence intensity of the experimental group and the control group of compounds 5a - 5e with the change of illumination time.

[0040] Figure 2 Trend chart of the fluorescence intensity of the experimental group and the control group of compounds 6a - 6e with the change of illumination time.

[0041] Figure 3 Trend chart of the fluorescence intensity of the experimental group and the control group of compounds 7a - 7e with the change of illumination time.

[0042] Figure 4 Trend chart of the fluorescence intensity of the experimental group and the control group of compounds 8a - 8f with the change of illumination time. Detailed Embodiments

[0043] The content of the present invention will be specifically described below through examples. In the present invention, the following examples are for better explaining the present invention and are not used to limit the scope of the present invention. The materials, reagents, etc. used in the following examples can be obtained from commercial channels without special instructions.

[0044] Example 1 Synthesis of Porphyrins 1, 2, and 3

[0045]

[0046] Pour the measured 20 mL of pyrrole into a 50 mL round-bottom flask, add a magnetic stir bar, place the round-bottom flask in an oil bath at 135 °C, successively assemble a distillation head, a thermometer, a straight condenser, and a cow horn tube. Insulate and shield the reaction system with a towel and aluminum foil, collect the fraction at 130 - 131 °C, and store it in a -20 °C refrigerator for subsequent reactions.

[0047] Add 2.44 g of p-hydroxybenzaldehyde, 6.12 mL of benzaldehyde / 8.4 g of p-chlorobenzaldehyde / 11.1 g of p-bromobenzaldehyde to a 500 mL three-necked flask, and then add 120 mL of propionic acid as a solvent and stir to dissolve at 135 °C. Add 5.5 mL of freshly distilled pyrrole to a constant pressure dropping funnel, mix it with an equal volume of propionic acid, and then slowly drop it into the reaction system, controlling the dropping to be completed within 20 - 30 min. Continue to reflux under condensation for 2 h. After the reaction solution is cooled to room temperature, pour 100 mL of absolute ethanol into the reaction solution, place the three-necked flask in a 4 °C refrigerator and refrigerate for 24 h. Filter by suction, wash the filter cake with absolute ethanol, and place it in a vacuum drying oven to dry to obtain a purple crude product. Purify by passing through a silica gel column using the developing agent dichloromethane: n-hexane = 3:1 to obtain porphyrin compounds 1 / 2 / 3.

[0048] Porphyrin compound 1: Yield: 8%. 1H NMR (500 MHz, Chloroform-d) δ 8.89–8.83 (m, 8H), 8.25–8.21 (m, 6H), 8.06 (d, J = 8.3 Hz, 2H), 7.80–7.73 (m, 9H), 7.16 (d, J = 8.3 Hz, 2H), -2.76 (s, 2H) ppm.

[0049] Porphyrin compound 2: Yield: 8%. 1H NMR (500 MHz, Chloroform-d) δ 8.92–8.81 (m, 8H), 8.14 (d, J = 7.8 Hz, 6H), 8.06 (d, J = 8.0 Hz, 2H), 7.75 (d, J = 7.8 Hz, 6H), 7.22 (d, J = 8.0 Hz, 2H), -2.84 (s, 2H) ppm.

[0050] Porphyrin compound 3: Yield: 7.4%. 1H NMR (500 MHz, Chloroform-d) δ 8.90–8.82 (m, 8H), 8.12 (d, J = 7.8 Hz, 6H), 8.03 (d, J = 8.3 Hz, 2H), 7.71 (d, J = 8.0 Hz, 6H), 7.33 (d, J = 8.5 Hz, 2H), -2.83 (s, 2H) ppm.

[0051] Example 2 Synthesis of formononetin derivatives 4a - 4e

[0052]

[0053] Add 268 mg of formononetin and 552.8 mg of anhydrous K 2 CO 3, 50 mL of acetone was added as a solvent, and the mixture was stirred and activated at 60 °C for 30 min. Then, 129.3 μL of 1,2-dibromoethane / 156.7 μL of 1,3-dibromopropane / 181.4 μL of 1,4-dibromobutane / 204.3 μL of 1,5-dibromopentane / 242.8 μL of 1,6-dibromohexane was slowly added dropwise to the reaction system. A thin-layer chromatography silica gel plate was used to monitor the progress of the reaction. After the reaction was completed, K 2 CO 3 was removed by suction filtration. The acetone was evaporated off using a rotary evaporator to obtain a pale yellow crude product. The product was purified by passing through a silica gel column chromatography using a developing solvent of dichloromethane:ethyl acetate = 30:1 to obtain formononetin derivatives 4a - 4e.

[0054] Compound 4a: Yield: 48%. 1H NMR (500 MHz, Chloroform-d) δ 8.22 (d, J = 8.8 Hz, 1H), 7.92 (s, 1H), 7.49 (d, J = 8.3 Hz, 2H), 7.02–6.95 (m, 3H), 6.85 (d, J = 2.4 Hz, 1H), 4.38 (t, J = 6.2 Hz, 2H), 3.83 (s, 3H), 3.69 (t, J = 6.2 Hz, 2H).

[0055] Compound 4b: Yield: 79%. 1H NMR (500 MHz, Chloroform-d) δ 8.42 (s, 1H), 8.04 (d, J = 8.9 Hz, 1H), 7.52 (d, J = 8.5 Hz, 2H), 7.19 (d, J = 2.2 Hz, 1H), 7.10 (dd, J = 9.0, 2.2 Hz, 1H), 6.99 (d, J = 8.5 Hz, 2H), 4.24 (t, J = 6.0 Hz, 2H), 3.79 (s, 3H), 3.69 (t, J = 6.5 Hz, 2H), 2.34–2.28 (m, 2H).

[0056] Compound 4c: Yield: 81%. 1H NMR (500 MHz, Chloroform-d) δ 8.21 (d, J = 8.8 Hz, 1H), 7.92 (s, 1H), 7.50 (d, J = 8.2 Hz, 2H), 6.97 (dd, J = 8.8, 2.8 Hz, 3H), 6.84 (d, J = 2.3 Hz, 1H), 4.10 (t, J = 5.9 Hz, 2H), 3.84 (s, 3H), 3.51 (t, J = 6.4 Hz, 2H), 2.13–1.99 (m, 4H).

[0057] Compound 4d: Yield: 77%. 1H NMR (500 MHz, Chloroform-d) δ 8.41 (s, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.52 (d, J = 8.4 Hz, 2H), 7.15 (d, J = 2.3 Hz, 1H), 7.07 (dd, J = 8.9, 2.3 Hz, 1H), 6.99 (d, J = 8.5 Hz, 2H), 4.13 (t, J = 6.4 Hz, 2H), 3.79 (s, 3H), 3.57 (t, J = 6.7 Hz, 2H), 1.93–1.75 (m, 4H), 1.60–1.53 (m, 2H).

[0058] Compound 4e: Yield: 70%. 1H NMR (500 MHz, Chloroform-d) δ 8.20 (d, J = 8.9 Hz, 1H), 7.91 (s, 1H), 7.50 (d, J = 8.3 Hz, 2H), 6.97 (d, J = 7.9 Hz, 3H), 6.83 (d, J = 2.3 Hz, 1H), 4.06 (t, J = 6.4 Hz, 2H), 3.84 (s, 3H), 3.44 (t, J = 6.7 Hz, 2H), 1.94–1.84 (m, 4H), 1.54–1.48 (m, 4H).

[0059] Example 3 Synthesis of Porphyrin-Formononetin of Formula I

[0060]

[0061] Add 94.6 mg of porphyrin compound 1 and 82.9 mg of anhydrous K 2 CO 3 to a 50 mL round-bottom flask, add 20 mL of DMF as the solvent, stir and activate at 60 °C for 30 min, and then add 37.4 mg of 4a / 38.8 mg of 4b / 40.2 mg of 4c / 41.6 mg of 4d / 43 mg of 4e to the reaction system. Use a chromatography silica gel plate to monitor the progress of the reaction. After the reaction is completed, remove K 2 CO 3 by suction filtration, extract with dichloromethane, wash the organic layer 4 times with water, remove water with anhydrous sodium sulfate, and spin off dichloromethane with a rotary evaporator to obtain a purple crude product. Purify through a chromatography silica gel column successively using the eluent dichloromethane, n-hexane:ethyl acetate = 6:1 to obtain porphyrin-formononetin derivatives 5a - 5e.

[0062]

[0063] Yield: 53.3%. 11H NMR (500 MHz, Chloroform-d) δ 8.89–8.83 (m, 8H), 8.30 (d, J = 8.9 Hz, 1H), 8.24–8.15 (m, 8H), 7.92 (s, 1H), 7.76 (d, J = 7.9 Hz, 9H), 7.53 (d, J = 8.8 Hz, 2H), 7.35 (d, J = 8.3 Hz, 2H), 7.17 (d, J = 8.9, 2.4 Hz, 1H), 7.06 (s, 1H), 6.99 (d, J = 9.0 Hz, 2H), 4.66 (d, J = 27.1 Hz, 4H), 3.86 (s, 3H), -2.78 (s, 2H). 13 13C NMR (126 MHz, Chloroform-d) δ 175.67, 163.10, 159.41, 158.40, 157.69, 151.67, 142.02, 142.89, 135.46, 134.66, 134.35, 129.94, 127.70, 127.60, 126.48, 124.73, 124.06, 119.90, 119.83, 119.71, 118.36, 114.59, 113.78, 112.66, 100.61, 65.10, 64.19, 55.14. HRMS-APCI: m / z calcd for C 62 H 44 N 4 O 5 + 925.3312 [M+H] + , found 925.3384.

[0064]

[0065] Yield: 69.3%. 1 1H NMR (500 MHz, Chloroform-d) δ 8.90–8.83 (m, 8H), 8.28–8.20 (m, 7H), 8.14 (d, J = 7.9 Hz, 2H), 7.92 (s, 1H), 7.80–7.73 (m, 9H), 7.50 (d, J = 8.7 Hz, 2H), 7.30 (d, J = 8.5 Hz, 2H), 7.11 (dd, J = 8.9, 2.4 Hz, 1H), 7.02–6.94 (m, 3H), 4.45 (dt, J = 6.9, 6.0 Hz, 4H), 3.85 (s, 3H), 2.57–2.49 (m, 2H), -2.77 (s, 2H). 1313C NMR (126 MHz, Chloroform-d) δ 175.68, 163.11, 159.42, 158.41, 157.80, 151.88, 142.03, 142.00, 135.46, 134.66, 134.36, 129.95, 127.71, 127.51, 126.49, 124.74, 124.07, 119.91, 119.83, 119.71, 118.37, 114.70, 113.79, 112.57, 100.62, 65.11, 64.20, 55.15, 29.12. HRMS-APCI: m / z calcd for C 63 H 46 N 4 O 5 + 939.3468 [M+H] + , found 939.3541.

[0066]

[0067] Yield: 77%. 1 1H NMR (500 MHz, Chloroform-d) δ 8.91–8.84 (m, 8H), 8.28–8.20 (m, 7H), 8.13 (d, J = 8.1 Hz, 2H), 7.91 (s, 1H), 7.82–7.71 (m, 9H), 7.49 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 7.07 (dd, J = 8.9, 2.3 Hz, 1H), 6.96 (d, J = 8.3 Hz, 2H), 6.92 (d, J = 2.2 Hz, 1H), 4.34 (t, J = 6.2 Hz, 2H), 4.26 (t, J = 6.4 Hz, 2H), 3.84 (s, 3H), 2.25–2.15 (m, 4H), -2.75 (s, 2H). 13 13C NMR (126 MHz, Chloroform-d) δ 175.87, 163.42, 159.61, 158.79, 157.99, 152.04, 142.25, 142.22, 135.66, 134.67, 134.57, 131.09, 130.14, 127.88, 127.72, 126.69, 124.90, 124.29, 120.11, 120.02, 118.47, 114.83, 113.98, 112.76, 100.75, 68.32, 67.65, 55.34, 26.14, 26.03. HRMS-APCI: m / z calcd for C64 H 48 N 4 O 5 + 953.3625 [M+H] + , found 953.3697.

[0068]

[0069] Yield: 81%. 1 HNMR (500 MHz, Chloroform-d) δ 8.92–8.83 (m, 8H), 8.25–8.20 (m, 7H), 8.12 (d, J = 8.0 Hz, 2H), 7.86 (s, 1H), 7.80–7.72 (m, 9H), 7.45 (d, J = 8.3 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 7.05 (dd, J = 8.9, 2.3 Hz, 1H), 6.92 (d, J = 8.3 Hz, 2H), 6.88 (d, J = 2.2 Hz, 1H), 4.29 (t, J = 6.2 Hz, 2H), 4.17 (t, J = 6.4 Hz, 2H), 3.82 (s, 3H), 2.10–2.02 (m, 4H), 1.90–1.83 (m, 2H), -2.75 (s, 2H). 13 C NMR (126 MHz, Chloroform-d) δ 175.86, 163.48, 159.58, 158.89, 157.98, 152.00, 142.25, 142.22, 135.65, 134.57, 130.11, 127.83, 127.71, 126.69, 124.87, 124.27, 120.10, 120.00, 118.40, 114.87, 113.95, 112.78, 100.68, 68.56, 67.99, 55.32, 29.20, 28.88, 22.91. HRMS-APCI: m / z calcd for C 65 H 50 N 4 O 5 + 967.3781 [M+H] + , found 967.3854.

[0070]

[0071] Yield: 87%. 1HNMR(500MHz, Chloroform-d) δ 8.90–8.83(m, 8H), 8.25–8.18(m, 7H), 8.12(d, J=8.1Hz, 2H), 7.86(s, 1H), 7.80–7.73(m, 9H), 7.45(d, J=8.6Hz, 2H), 7.27(d, J=8.3Hz, 2H), 7.03(dd, J,=8.9, 2.3Hz, 1H), 6.92(d, J=8.5Hz, 2H), 6.88(d, J=2.3Hz, 1H), 4.28(t, J=6.3Hz, 2H), 4.14(t, J=6.4Hz, 2H), 3.82(s, 3H), 2.03–1.95(m, 4H), 1.78–1.67(m, 4H), -2.75(s, 2H). 13 C NMR(126MHz, Chloroform-d) δ 175.86, 163.53, 159.57, 158.96, 157.98, 152.00, 142.26, 142.22, 135.65, 134.57, 134.49, 130.10, 127.80, 127.70, 126.68, 124.85, 124.28, 120.09, 119.98, 118.37, 114.84, 113.95, 112.78, 100.69, 68.61, 68.11, 55.32, 29.43, 29.04, 26.04, 25.93. HRMS-APCI: m / z calcd for C 66 H 52 N 4 O 5 + 981.3938[M + H] + , found 981.1040.

[0072] Add 110 mg of porphyrin compound 2 and 82.9 mg of anhydrous K 2 CO 3 , add 20 mL of DMF as the solvent, stir and activate at 60 °C for 30 min, and then add 37.4 mg of 4a / 38.8 mg of 4b / 40.2 mg of 4c / 41.6 mg of 4d / 43 mg of 4e to the reaction system. Use a chromatography silica gel plate to spot-check and monitor the progress of the reaction. After the reaction is completed, remove K 2 CO 3Removed by suction filtration, extracted with dichloromethane, the organic layer was washed with water 4 times, dried over anhydrous sodium sulfate, and the dichloromethane was removed by rotary evaporation to obtain a purple crude product. Purified by passing through a silica gel column for chromatography successively using the eluents dichloromethane, dichloromethane:ethyl acetate = 30:1 to obtain porphyrin-formononetin derivatives 6a - 6e.

[0073]

[0074] Yield: 60%. 1 H NMR (500 MHz, Chloroform-d) δ 8.90–8.82 (m, 8H), 8.29 (d, J = 8.8 Hz, 1H), 8.16–8.11 (m, 8H), 7.95 (s, 1H), 7.74 (d, J = 8.1 Hz, 6H), 7.53 (d, J = 8.7 Hz, 2H), 7.33 (d, J = 8.5 Hz, 2H), 7.15 (dd, J = 8.9, 2.4 Hz, 1H), 7.03–6.97 (m, 3H), 4.63–4.57 (m, 4H), 3.86 (s, 3H), -2.83 (s, 2H). 13 C NMR (126 MHz, Chloroform-d) δ 175.86, 162.99, 159.67, 158.42, 157.89, 152.12, 140.51, 140.46, 135.65, 135.51, 134.95, 134.36, 130.16, 128.05, 127.02, 125.01, 124.20, 120.37, 118.84, 118.65, 114.80, 114.03, 113.00, 101.15, 67.26, 66.50, 55.36. HRMS-APCI: m / z calcd for C 62 H 41 Cl 3 N 4 O 5 + 1027.2143 [M + H] + , found 1027.2215.

[0075]

[0076] Yield: 71%. 11H NMR (500 MHz, Chloroform-d) δ 8.92–8.78 (m, 8H), 8.27 (d, J = 8.9 Hz, 1H), 8.19–8.05 (m, 8H), 7.92 (s, 1H), 7.78–7.70 (m, 6H), 7.50 (d, J = 8.7 Hz, 2H), 7.31 (d, J = 8.5 Hz, 2H), 7.09 (dd, J = 8.9, 2.4 Hz, 1H), 7.00–6.94 (m, 3H), 4.49–4.41 (m, 4H), 3.85 (s, 3H), 2.55–2.50 (m, 2H), -2.84 (s, 2H). 13 13C NMR (126 MHz, Chloroform-d) δ 175.86, 163.29, 159.63, 158.74, 157.98, 152.06, 140.52, 140.48, 135.66, 135.50, 134.53, 134.34, 130.14, 127.93, 127.03, 127.01, 124.95, 124.24, 120.55, 118.80, 118.59, 114.86, 114.00, 112.86, 100.83, 65.27, 64.42, 55.35, 29.33. HRMS-APCI: m / z calcd for C 63 H 43 Cl 3 N 4 O 5 + 1041.2299 [M+H] + , found 1041.2371.

[0077]

[0078] Yield: 73%. 1 1H NMR (500 MHz, Chloroform-d) δ 8.93–8.80 (m, 8H), 8.25 (d, J = 8.9 Hz, 1H), 8.16–8.12 (m, 8H), 7.91 (s, 1H), 7.77–7.71 (m, 6H), 7.50 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.2 Hz, 2H), 7.07 (dd, J = 8.9, 2.4 Hz, 1H), 6.99–6.91 (m, 3H), 4.36 (t, J = 6.3 Hz, 2H), 4.26 (t, J = 6.4 Hz, 2H), 3.84 (s, 3H), 2.27–2.19 (m, 4H), -2.83 (s, 2H). 1313C NMR (126 MHz, Chloroform-d) δ 175.83, 163.46, 159.55, 159.03, 157.97, 151.99, 140.51, 140.49, 135.66, 135.50, 134.31, 134.22, 130.09, 127.85, 127.00, 126.01, 124.85, 124.24, 120.70, 118.77, 118.52, 118.41, 114.84, 113.90, 112.87, 100.65, 68.58, 68.09, 55.35, 28.76, 28.25. HRMS-APCI: m / z calcd for C 64 H 45 Cl 3 N 4 O 5 + 1055.2456 [M+H] + , found 1055.2528.

[0079]

[0080] Yield: 70%. 1 1H NMR (500 MHz, Chloroform-d) δ 8.93–8.87 (m, 8H), 8.24 (d, J = 9.1 Hz, 1H), 8.19–8.05 (m, 8H), 7.87 (s, 1H), 7.74 (d, J = 7.4 Hz, 6H), 7.44 (d, J = 7.2 Hz, 2H), 7.29 (d, J = 7.1 Hz, 2H), 7.05 (d, J = 8.5 Hz, 1H), 6.96–6.85 (m, 3H), 4.31 (t, J = 6.0 Hz, 2H), 4.19 (t, J = 6.1 Hz, 2H), 3.82 (s, 3H), 2.15–2.08 (m, 4H), 1.91–1.85 (m, 2H), -2.82 (s, 2H). 13CNMR(126MHz, Chloroform-d) δ 175.85, 163.47, 159.58, 159.02, 157.98, 152.00, 140.54, 140.49, 135.65, 135.51, 134.33, 134.23, 130.09, 127.84, 127.03, 127.01, 124.87, 124.24, 120.72, 118.78, 118.54, 118.41, 114.85, 113.94, 112.86, 100.69, 68.56, 68.02, 55.32, 29.20, 28.89, 22.92. HRMS-APCI: m / z calcd for C 64 H 47 Cl 3 N 4 O 5 + 1069.2612 [M+H] + , found 1069.2684.

[0081]

[0082] Yield: 88%. 1 H NMR(500MHz, Chloroform-d) δ 8.99–8.72(m, 8H), 8.23(d, J=8.9Hz, 1H), 8.15–8.12(m, 8H), 7.85(s, 1H), 7.79–7.69(m, 6H), 7.44(d, J=8.4Hz, 2H), 7.29(d, J=8.2Hz, 2H), 7.03(dd, J=8.9, 2.3Hz, 1H), 6.92(d, J=8.4Hz, 2H), 6.88(d, J=2.4Hz, 1H), 4.28(t, J=6.3Hz, 2H), 4.15(t, J=6.4Hz, 2H), 3.82(s, 3H), 2.07–1.95(m, 4H), 1.79–1.70(m, 4H), -2.83(s, 2H). 1313C NMR (126 MHz, Chloroform-d) δ 175.64, 163.31, 159.37, 158.89, 157.77, 151.78, 140.34, 140.29, 135.45, 135.31, 134.13, 133.96, 129.89, 127.60, 126.83, 126.81, 124.64, 124.05, 120.56, 118.58, 118.34, 118.16, 114.63, 113.74, 112.66, 100.47, 68.40, 67.93, 55.12, 29.51, 29.23, 25.85, 25.74. HRMS-APCI: m / z calcd for C 65 H 49 Cl 3 N 4 O 5 + 1083.2769 [M+H] + , found 1083.2841.

[0083] 86.44 mg of porphyrin compound 3 and 82.9 mg of anhydrous K 2 CO 3 were added to a 50 mL round-bottom flask. 20 mL of DMF was added as a solvent, and the mixture was stirred and activated at 60 °C for 30 min. Then 37.4 mg of 4a / 38.8 mg of 4b / 40.2 mg of 4c / 41.6 mg of 4d / 43 mg of 4e were added to the reaction system. The progress of the reaction was monitored by spotting on a chromatographic silica gel plate. After the reaction was completed, K 2 CO 3 was removed by suction filtration, and the mixture was extracted with dichloromethane. The organic layer was washed with water 4 times, dried over anhydrous sodium sulfate, and the dichloromethane was removed by rotary evaporation to obtain a purple crude product. The product was purified by passing through a chromatographic silica gel column successively using dichloromethane and dichloromethane:ethyl acetate = 40:1 as eluents to obtain porphyrin-formononetin derivatives 7a - 7e.

[0084]

[0085] Yield: 49%. 11H NMR (500 MHz, Chloroform-d) δ 8.92–8.80 (m, 8H), 8.30 (d, J = 8.9 Hz, 1H), 8.13 (d, J = 8.3 Hz, 2H), 8.10–8.07 (m, 6H), 7.96 (s, 1H), 7.90 (d, J = 8.0 Hz, 6H), 7.53 (d, J = 8.7 Hz, 2H), 7.33 (d, J = 8.5 Hz, 2H), 7.16 (dd, J = 8.9, 2.4 Hz, 1H), 7.03–6.98 (m, 3H), 4.66–4.59 (m, 4H), 3.86 (s, 3H), -2.84 (s, 2H). 13 13C NMR (126 MHz, Chloroform-d) δ 175.85, 163.00, 159.67, 158.44, 157.89, 152.11, 140.98, 140.93, 135.84, 135.65, 134.94, 130.16, 129.97, 128.06, 125.01, 124.21, 122.59, 120.41, 118.86, 118.82, 118.63, 114.79, 114.03, 113.01, 101.16, 67.27, 66.52, 55.36. HRMS-APCI: m / z calcd for C 62 H 41 Br 3 N 4 O 5 + 1161.0607 [M+H] + , found 1161.0645.

[0086]

[0087] Yield: 73%. 1 1H NMR (500 MHz, Chloroform-d) δ 8.93–8.79 (m, 8H), 8.27 (d, J = 8.8 Hz, 1H), 8.13–8.05 (m, 8H), 7.94–7.87 (m, 7H), 7.50 (d, J = 8.6 Hz, 2H), 7.31 (d, J = 8.3 Hz, 2H), 7.10 (dd, J = 8.8, 2.3 Hz, 1H), 6.99–6.94 (m, 3H), 4.50–4.45 (m, 4H), 3.85 (s, 3H), 2.55–2.50 (m, 2H), -2.85 (s, 2H). 1313C NMR (126 MHz, Chloroform-d) δ 175.85, 163.29, 159.64, 158.75, 157.98, 152.05, 141.00, 140.95, 135.84, 135.65, 134.51, 130.14, 129.98, 129.96, 127.93, 124.95, 124.25, 122.58, 120.59, 118.79, 118.59, 118.57, 114.85, 114.00, 112.87, 100.84, 65.28, 64.43, 55.35, 53.40, 29.33. HRMS-APCI: m / z calcd for C 63 H 43 Br 3 N 4 O 5 + 1175.0763 [M+H] + , found 1175.3384.

[0088]

[0089] Yield: 77%. 1 1H NMR (500 MHz, Chloroform-d) δ 8.91–8.81 (m, 8H), 8.26 (d, J = 8.9 Hz, 1H), 8.05–7.94 (m, 8H), 7.93–7.89 (m, 7H), 7.50 (d, J = 8.4 Hz, 2H), 7.30 (d, J = 8.2 Hz, 2H), 7.07 (dd, J = 8.8, 2.3 Hz, 1H), 6.97–6.93 (m, 3H), 4.37 (t, J = 6.3 Hz, 2H), 4.28 (t, J = 6.4 Hz, 2H), 3.84 (s, 3H), 2.27–2.19 (m, 4H), -2.84 (s, 2H). 13 13C NMR (126 MHz, Chloroform-d) δ 175.83, 163.45, 159.62, 159.03, 157.95, 151.94, 141.02, 140.99, 135.79, 135.63, 134.20, 130.11, 129.97, 129.95, 127.88, 124.79, 124.23, 122.56, 120.76, 118.81, 118.52, 118.41, 114.87, 113.92, 112.87, 100.71, 68.55, 68.05, 55.31, 29.22, 28.87. HRMS-APCI: m / z calcd for C64 H 45 Br 3 N 4 O 5 + 1189.0920[M+H] + , found 1189.0961.

[0090]

[0091] Yield: 81%. 1 H NMR (500 MHz, Chloroform-d) δ 8.93–8.81 (m, 8H), 8.24 (d, J = 8.9 Hz, 1H), 8.13–8.05 (m, 8H), 7.92–7.86 (m, 7H), 7.44 (d, J = 8.6 Hz, 2H), 7.29 (d, J = 8.3 Hz, 2H), 7.05 (dd, J = 8.9, 2.4 Hz, 1H), 6.92 (d, J = 8.5 Hz, 2H), 6.89 (d, J = 2.3 Hz, 1H), 4.30 (t, J = 6.2 Hz, 2H), 4.19 (t, J = 6.4, 2H), 3.82 (s, 3H), 2.12–2.04 (m, 4H), 1.92–1.85 (m, 2H), -2.84 (s, 2H). 13 C NMR (126 MHz, Chloroform-d) δ 175.83, 163.47, 159.58, 159.03, 157.98, 151.98, 141.02, 140.97, 135.84, 135.65, 134.21, 130.09, 129.98, 129.95, 127.84, 124.87, 124.25, 122.57, 120.76, 118.77, 118.53, 118.42, 114.84, 113.94, 112.87, 100.69, 68.56, 68.02, 55.31, 29.20, 28.89, 22.92. HRMS-APCI: m / z calcd for C 65 H 47 Br 3 N 4 O 5 + 1203.1076[M+H] + , found 1203.1101.

[0092]

[0093] Yield: 88%. 11H NMR (500 MHz, Chloroform-d) δ 8.92–8.81 (m, 8H), 8.23 (d, J = 8.9 Hz, 1H), 8.12–8.06 (m, 8H), 7.91–7.88 (m, 6H), 7.86 (s, 1H), 7.44 (d, J = 8.6 Hz, 2H), 7.29 (d, J = 8.4 Hz, 2H), 7.03 (dd, J = 8.9, 2.4 Hz, 1H), 6.92 (d, J = 8.6 Hz, 2H), 6.88 (d, J = 2.4 Hz, 1H), 4.28 (t, J = 6.3 Hz, 2H), 4.15 (t, J = 6.4 Hz, 2H), 3.82 (s, 3H), 2.06–1.97 (m, 4H), 1.77–1.70 (m, 4H), -2.84 (s, 2H). 13 13C NMR (126 MHz, Chloroform-d) δ 175.84, 163.51, 159.57, 157.97, 151.97, 141.02, 140.97, 135.84, 135.64, 134.14, 130.08, 129.98, 129.95, 127.81, 124.85, 124.25, 122.56, 120.80, 118.76, 118.52, 118.37, 114.83, 114.00, 113.94, 112.86, 100.69, 68.61, 68.14, 55.32, 29.43, 29.04, 26.05, 25.94. HRMS-APCI: m / z calcd for C 66 H 49 Br 3 N 4 O 5 + 1217.1233 [M+H] + , found 1217.1246.

[0094] Example 4 Synthesis of Porphyrin-Formononetin of Formula II

[0095]

[0096] Add 48.3 mg of 5d / 49 mg of 5e / 53.4 mg of 6d / 54.1 mg of 6e / 60 mg of 7d / 60.7 to a 50 mL round-bottom flask

[0097] mg7e, 43.9 mg of zinc acetate dihydrate, 20 mL of dichloromethane was added as a solvent, and the reaction was stirred at 40 °C. The progress of the reaction was monitored by spotting on a chromatography silica gel plate. After the reaction was completed, the unreacted zinc acetate was removed by suction filtration, the filtrate was washed with distilled water, dried over anhydrous sodium sulfate, and the dichloromethane was removed by rotary evaporation to obtain a purple-red crude product. The product was purified by passing through a chromatography silica gel column using dichloromethane as the eluent to obtain the metallated compound of porphyrin-formononetin 8a - 8f.

[0098]

[0099] Yield: 91%. 1 H NMR (500 MHz, Chloroform - d) δ 9.00–8.93 (m, 8H), 8.24–8.17 (m, 7H), 8.12 (d, J = 8.0 Hz, 2H), 7.85 (s, 1H), 7.80–7.74 (m, 9H), 7.41 (d, J = 8.3 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 7.03 (dd, J = 9.0, 2.2 Hz, 1H), 6.92–6.84 (m, 3H), 4.31 (t, J = 6.3 Hz, 2H), 4.19 (t, J = 6.4 Hz, 2H), 3.80 (s, 3H), 2.11–2.03 (m, 4H), 1.91–1.85 (m, 2H). 13 C NMR (126 MHz, Chloroform - d) δ 175.87, 163.48, 159.69, 158.90, 157.99, 152.02, 142.27, 142.23, 140.05, 135.67, 134.68, 130.11, 127.83, 127.71, 126.69, 124.87, 124.27, 120.10, 120.00, 118.40, 114.87, 113.95, 112.78, 100.68, 68.67, 68.00, 55.32, 29.22, 28.88, 22.93. HRMS - APCI: m / z calcd for C 65 H 48 N 4 O 5 Zn + 1029.2916 [M + H] + , found 1029.2988.

[0100]

[0101] Yield: 95%. 1HNMR (500 MHz, Chloroform-d) δ 8.93–8.85 (m, 8H), 8.25–8.21 (m, 7H), 8.11 (d, J=7.9 Hz, 2H), 7.85 (s, 1H), 7.81–7.75 (m, 9H), 7.42 (d, J=8.2 Hz, 2H), 7.03 (d, J=9.0 Hz, 1H), 6.89 (d, J=9.8 Hz, 3H), 4.29 (t, J=6.0 Hz, 2H), 4.15 (t, J=6.2 Hz, 2H), 3.80 (s, 3H), 2.07–1.97 (m, 4H), 1.80–1.67 (m, 4H). 13 C NMR (126 MHz, Chloroform-d) δ 175.87, 163.63, 159.67, 158.97, 157.99, 152.00, 142.26, 142.23, 140.67, 135.65, 134.67, 134.50, 130.10, 127.81, 127.71, 126.69, 124.86, 124.28, 120.09, 119.98, 118.37, 114.84, 113.95, 112.78, 100.69, 68.61, 68.12, 55.32, 29.44, 29.04, 26.05, 25.91. HRMS-APCI: m / z calcd for C 66 H 50 N 4 O 5 Zn + 1043.3073 [M+H] + , found 1043.3145.

[0102]

[0103] Yield: 89%. 1 HNMR (500 MHz, Chloroform-d) δ 9.11–8.90 (m, 8H), 8.20–8.09 (m, 9H), 7.83 (s, 1H), 7.74 (s, 6H), 7.37 (d, J=8.2 Hz, 2H), 7.00 (d, J=7.4 Hz, 1H), 6.90–6.84 (m, 3H), 4.31 (t, J=6.0 Hz, 2H), 4.19 (t, J=6.1 Hz, 2H), 3.79 (s, 3H), 2.13–2.06 (m, 4H), 1.92–1.83 (m, 2H). 13¹³C NMR (126 MHz, Chloroform-d) δ 175.84, 163.47, 159.67, 159.02, 157.97, 151.69, 140.64, 140.49, 139.89, 135.65, 135.50, 134.33, 134.22, 130.04, 127.83, 127.03, 127.00, 124.87, 124.24, 120.72, 118.78, 118.64, 118.41, 114.85, 113.94, 112.85, 100.68, 68.66, 68.01, 55.32, 29.20, 28.89, 22.92. HRMS-APCI: m / z calcd for C 65 H 45 Cl 3 N 4 O 5 Zn + 1131.1747 [M+H] + , found 1131.1756.

[0104]

[0105] Yield: 93%. 1 ¹H NMR (500 MHz, Chloroform-d) δ 9.04–8.88 (m, 8H), 8.15–8.07 (m, 9H), 7.82 (s, 1H), 7.78–7.68 (m, 6H), 7.36 (d, J = 8.6 Hz, 2H), 7.00 (dd, J = 8.9, 2.4 Hz, 1H), 6.90–6.83 (m, 3H), 4.29 (t, J = 6.3 Hz, 2H), 4.14 (t, J = 6.4 Hz, 2H), 3.79 (s, 3H), 2.06–1.98 (m, 4H), 1.78–1.68 (m, 4H). 1313C NMR (126 MHz, Chloroform-d) δ 175.63, 163.30, 159.36, 159.88, 157.76, 151.77, 140.34, 140.28, 139.66, 135.44, 135.30, 134.12, 133.95, 129.88, 127.60, 126.82, 126.80, 124.63, 124.04, 120.66, 118.67, 118.33, 118.15, 114.62, 113.73, 112.65, 100.46, 68.39, 67.92, 55.11, 29.22, 28.63, 26.82, 26.74. HRMS-APCI: m / z calcd for C 66 H 47 Cl 3 N 4 O 5 Zn + 1145.1904 [M+H] + , found 1145.1976.

[0106]

[0107] Yield: 88%. 1 1H NMR (500 MHz, Chloroform-d) δ 9.09–8.91 (m, 8H), 8.25 (d, J = 8.9 Hz, 1H), 8.13–7.99 (m, 8H), 7.89–7.82 (m, 7H), 7.36 (d, J = 8.3 Hz, 2H), 7.00 (dd, J = 9.1, 2.2 Hz, 1H), 6.89–6.81 (m, 3H), 4.30 (t, J = 6.4 Hz, 2H), 4.16 (t, J = 6.3 Hz, 2H), 3.79 (s, 3H), 2.14–2.01 (m, 4H), 1.95–1.86 (m, 2H). 1313C NMR(126MHz,Chloroform-d)δ175.82,163.46,159.67,159.02,157.97,151.91,141.01,140.98,140.36,135.83,135.64,134.20,130.09,129.97,129.94,127.83,124.86,124.24,122.66,120.76,118.76118.62,118.41,114.83,113.94,112.86,100.68,68.66,68.01,55.31,29.20,28.89,22.92.HRMS-APCI:m / z calcdfor C 65 H 45 Br 3 N 4 O 5 Zn + 1265.0211[M+H] + ,found 1265.0420.

[0108]

[0109] Yield: 88%. 1 1H NMR(500MHz,Chloroform-d)δ8.98–8.72(m,8H),8.15–8.07(m,9H),7.83(s,1H),7.74(s,6H),7.37(d,J = 8.1Hz,2H),6.99(d,J = 9.0Hz,1H),6.89–6.84(m,3H),4.29(s,2H),4.13(t,J = 6.5Hz,2H),3.79(s,3H),2.06–1.95(m,4H),1.78–1.65(m,4H). 13 13C NMR(126MHz,Chloroform-d)δ175.83,163.47,159.61,157.98,151.96,141.02,140.97,140.45,135.83,135.58,134.14,130.13,129.86,129.95,127.87,124.85,124.35,122.56,120.76,118.78,118.52,118.41,114.88,114.00,113.94,112.86,100.69,68.67,68.14,55.32,29.45,29.05,26.06,25.98.HRMS-APCI:m / z calcd for C66 H 47 Br 3 N 4 O 5 Zn + 1279.0368[M+H] + , found 1279.0418。

[0110] Example 4 Singlet Oxygen Detection Experiment

[0111] Precisely weigh 10 μmol of DPBF and the compound to be tested using an electronic balance, add 1 mL of chloroform to each, and prepare the sample mother liquor. Then dilute with chloroform to a 20 μmol / L DPBF solution and a 5 μmol / L solution of the compound to be tested. Mix DPBF and the compound to be tested in equal volumes. Use a 1-cm glass cell, set the excitation wavelength to 418 nm, the scanning speed to 2400 nm / min, and the scanning range to 200–800 nm. Measure the change in the fluorescence intensity of DPBF in the glass cell at 0 s, 10 s, 20 s, 30 s, 40 s, and 50 s of illumination. Conduct three parallel experiments for each group.

[0112] Analysis based on the change in the fluorescence intensity of DPBF at 463 nm at 0 s, 10 s, 20 s, 30 s, 40 s, and 50 s of illumination shows that the fluorescence intensity of DPBF itself only changed slightly within 50 s of illumination, and this change can be ignored. When formononetin was present, the fluorescence intensity of DPBF decreased to a certain extent. The porphyrin compounds (1, 2, 3) and 5a - 8f caused a significant decrease in the fluorescence intensity of DPBF at 10 s of illumination, with the fluorescence intensity decreasing by more than 90%. This indicates that a large amount of ROS was generated to destroy the cyclic structure of DPBF, resulting in the fluorescence quenching of DPBF. The fluorescence intensity of compounds 5a - 8f decreased more than that of their corresponding porphyrin precursors, which indicates that compounds 5a - 8f generated more ROS than their corresponding porphyrin precursors. The reason may be that formononetin also has the ability to generate ROS, so that after the porphyrin precursor combines with formononetin, they can synergistically generate ROS.

[0113] For the fluorescence change diagrams of the target compounds 5a - 8f and the control group, see the appendix Figures 1-4 . In the determination of the ultraviolet - visible absorption spectrum, compounds 5a - 8f all contain the Soret band and Q band of the starting porphyrin and the characteristic absorption peak of formononetin at around 250 nm. According to whether Zn is inserted into the center of the porphyrin ring 2+ , the number of Q bands shows a difference. When Zn is inserted 2+When the time comes, due to the increased symmetry of the molecular structure and the increased degeneracy of molecular orbitals, the number of Q bands decreases. In addition, due to the different substituents in the structure, the characteristic absorption peaks in compounds 5a - 8f will show a red shift or a blue shift relative to the raw materials. In the determination of ROS, as mentioned above, formononetin causes a certain degree of decrease in the fluorescence intensity of DPBF. Compared with the two raw materials, compounds 5a - 8f cause a significant decrease in the fluorescence intensity of DPBF after illumination, indicating that the porphyrin-formononetin compounds generate more ROS after illumination.

[0114] Example 5 Detection of in vitro anti-tumor cell activity by MTT colorimetric method

[0115] The inhibitory activities of the target compounds and control compounds against different cells were determined by the MTT method. A blank control group, a negative control group, a positive control group, and an experimental group were set up. Among them, DMSO was used as the negative control group, 5-FU and Ce6 were used as the positive control group, the raw material formononetin (FMN), porphyrin compounds (1, 2, 3), and the target compounds 5a - 8f were used as the experimental group. Both the blank control group and the negative control group were set with 6 replicates, and the positive control group and the experimental group were set with 3 replicates.

[0116] Table 1 Anti-A549 and MDA-MB-231 cell activities of porphyrin-formononetin derivatives

[0117]

[0118]

[0119]

[0120] Table 2 Anti-HCT-116 and HGC-27 cell activities of porphyrin-formononetin derivatives

[0121]

[0122]

[0123] Table 3 Anti-DU145, TCCSUP, and H9c2 cell activities of porphyrin-formononetin derivatives

[0124]

[0125]

[0126] As can be seen from Table 1, Table 2 and Table 3, the porphyrin-formononetin derivatives of the present invention show varying degrees of inhibitory effects on the growth of six cancer cells under light and dark conditions, while most of the compounds have little toxicity to normal cells. Especially under light conditions, the porphyrin-formononetin derivatives have better growth inhibitory effects on cancer cells compared to the porphyrin parent and formononetin raw materials. This is because ROS is generated after porphyrin is irradiated with light, which damages the functional structure of cancer cells, thereby inhibiting their growth and reproduction. In addition, formononetin, as a natural anti-cancer compound, itself also has an inhibitory effect on the growth of cancer cells. Therefore, under light conditions, the synergistic effects of phototherapy and chemotherapy are exhibited.

Claims

1. A porphyrin-formononetin derivative and a pharmaceutically acceptable salt thereof, which has a structure shown in Formula I or Formula II: in, n is an integer selected from 1 to 8; R is independently selected from H, halogen, hydroxy, nitro, CN, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 2-10 heteroaryl; R' is selected from H, halogen, hydroxy, nitro, CN, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl; M is selected from Zn, Ni, and Mn.

2. The porphyrin-formononetin derivative and its pharmaceutically acceptable salt according to claim 1, characterized in that: n is selected from 1, 2, 3, 4 or 5. R is selected from H, halogen, hydroxy, nitro, CN, C1-6 alkyl; preferably, R is selected from H or Cl; R' is selected from H, halogen, hydroxy, nitro, CN, C1-6 alkyl; preferably, R is selected from H; M is selected from Zn.

3. A method for preparing the compound of formula I as claimed in claim 1, wherein the reaction scheme is as follows: in, n, R, and R' are as defined in claim 1. The specific reaction steps are as follows: add a base and porphyrin 1 to an organic solvent, heat and stir, add the formononetin derivative 2 to the reaction system until the reaction is completed, and obtain the porphyrin-formononetin derivative of formula I through post-treatment.

4. The preparation method according to claim 3, characterized in that: The molar ratio of the porphyrin derivative 1 to the formononetin derivative 2 is: 1:(1-1.5); the base is selected from potassium hydroxide, triethylamine or potassium carbonate, more preferably potassium carbonate.

5. A method for preparing the compound of formula II as claimed in claim 1, wherein the reaction scheme is as follows: in, n, R, R' and M are as defined in claim 1; The specific reaction steps are as follows: dissolving the compound of formula I in an organic solvent, adding M metal salt, reflux reaction, monitoring by TLC until the reaction is complete, and obtaining the porphyrin-formononetin derivative of formula II through post-treatment.

6. The preparation method according to claim 5, characterized in that: The molar ratio of the compound of formula I to the metal salt of M is 1:3-7, preferably 1:

5.

7. A pharmaceutical composition comprising the porphyrin-forononetin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2, and a pharmaceutically acceptable carrier.

8. Use of the porphyrin-forononetin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2 or the pharmaceutical composition according to claim 7 in the preparation of a drug for treating cancer.

9. The method of claim 8, wherein the cancer is selected from the group consisting of lung cancer, breast cancer, colon cancer, stomach cancer, prostate cancer, and bladder cancer.

Images