A no-type porphyrin-ferulic acid derivative, a preparation method and use thereof

Abstract

The application discloses a NO type porphyrin-ferulic acid derivative, which has excellent singlet oxygen generation capacity and can effectively release NO. Through related researches on photodynamic properties and in-vitro anti-tumor activity, an anti-tumor drug with good synergistic photodynamic-chemotherapy treatment effect is obtained. Under the condition of illumination, the compound has good inhibition effect on tumor cells. Therefore, the compound can be used as an active ingredient of an anti-tumor drug and has good development and application prospect.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a NO-type porphyrin-ferulic acid derivative, its preparation method, and its uses. Background Technology

[0002] Photodynamic therapy (PDT) was discovered over 100 years ago and has become an widely investigated therapy for treating cancer and various non-malignant diseases, including infections. To date, this therapy has been successfully used to treat a variety of malignant tumors. The basic elements of PDT can be divided into: photosensitizer (PS), specific excitation light, and molecular oxygen. The photosensitizer, when irradiated with light of appropriate energy wavelength, forms an excited singlet state, which then transforms into a long-lived excited triplet state. This triplet state can undergo a photochemical reaction in the presence of oxygen, transferring energy to surrounding oxygen molecules to form reactive oxygen species (ROS), thereby killing cancer cells, pathogens, and unwanted tissues. Compared to more traditional cancer therapies such as surgery, radiation therapy, and drug therapy, PDT has advantages such as being non-invasive and having fewer side effects, making it highly attractive in the field of tumor treatment research.

[0003] With the development of photodynamic therapy (PDT), the preparation of photosensitizers, especially porphyrin-based photosensitizers, has made remarkable progress. Most photosensitizers used in cancer treatment have a macrocyclic skeleton based on porphyrins. The main advantages of porphyrin compounds in photodynamic research include: 1) the stability of aromatic compounds; 2) efficient absorption of visible light; 3) high reactive oxygen species yield; 4) ease of functionalization and structural diversity; and 5) long triplet lifetime and low dark toxicity. Photofri, the first photosensitizing drug approved by the US FDA, 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 its in vitro anticancer activity against human liver cancer cells SMCC-7721 using the MTT assay. Preliminary results showed that the anticancer activity of the coupled compounds was more than twice that of 5-fluorouracil. Porphyrin had no killing effect on tumor cells under light-free conditions, indicating that the coupled compounds actually enhanced the targeting of 5-fluorouracil to tumor tissues, thereby enhancing the anticancer activity. S. Weimin et al. (Bioorg Med Chem, 2008, 16(10): 5665-5671) synthesized a series of porphyrin compounds containing 5-fluorouracil / L-phenylalanine by coupling 5-fluorouric acid / Boc-L-phenylalanine with aminoporphyrin. In vitro anticancer activity studies showed that the introduction of 5-fluorouracil and L-phenylalanine significantly enhanced the phototoxicity of porphyrins. The high selectivity of porphyrin compounds for tumor tissues broadens the prospects for research on porphyrin-linked anticancer drugs.

[0005] Currently, ferulic acid (FA) and its sodium salt have been gradually shown to have good anti-tumor activity, and have been shown to have anti-cancer activity against human renal adenocarcinoma (ACHN), human bladder cancer (T24), human breast cancer (MDA-MB-231), and human osteosarcoma (143B and MG63) cells. Ferulic acid has different mechanisms of action against different cancer cell lines, including altering the cell cycle, inducing apoptosis, and regulating protein production. For example, Nasr Bouzaiene N et al. (Eur J Pharmacol. 2015, 766: 99-105) reported that FA inhibits the proliferation of A549 cell line by inhibiting DNA synthesis, and at higher concentrations, it can reduce the survival rate of A549 cells to 23%. Janicke et al. (J Agric Food Chem. 2005, 53(17): 6658-6665) showed that FA can inhibit the proliferation of human colonic endothelial tumor cells by affecting the cell cycle. Luo et al. (Med Sci Monit, 2020, 26:e920095) treated Caski cells with ferulic acid and found that ferulic acid downregulated the PI3K / Akt signaling pathway in Caski cells and induced Caski cell apoptosis.

[0006] Because ferulic acid has easily modifiable hydroxyl and carboxyl groups in its structure, it is readily modified to obtain a series of ferulic acid derivatives. In recent years, an increasing number of ferulic acid derivatives have been reported, exhibiting higher antitumor activity and stability than ferulic acid itself. Yue et al. modified the carboxyl group in the ferulic acid structure to obtain the ferulic acid derivative FXS-3. Experimental results showed that FXS-3 could induce apoptosis in A549 cells and arrest their cell cycle at the G0 / G1 phase. Sawata et al. linked two ferulic acid monomers with resveratrol to obtain the novel compound UHA025. Compared with resveratrol monomers and resveratrol linked only to one ferulic acid monomer, UHA025 exerted a stronger inhibitory effect on the 3D proliferation of HCT-116 cells by increasing the mRNA level of the tumor suppressor factor p15. Pellerito et al. synthesized tributyltin (IV) ferulic acid TBT-F, which can increase cell membrane permeability by arresting the G2 / M cell cycle.

[0007] Nitric oxide (NO) is one of the simplest and most widely used endogenous molecules, capable of directly influencing many biological processes. When NO reaches high concentrations at tumor sites, it has been shown to be an effective anticancer agent by inhibiting tumor growth through mechanisms such as suppressing mitochondrial respiration, affecting tumor cell cycle arrest, and producing tumor toxicity. Furthermore, NO can promote intracellular GSH metabolism, disrupt intracellular redox balance, and increase intracellular ROS levels. Notably, NO can also react with ROS to produce more potent cytotoxic peroxynitrite anions (ONOO-) and other reactive nitrogen species (RNS), leading to apoptosis, inflammation, DNA deamination, cellular function suppression, and mitochondrial respiratory damage through nitrosation and oxidative stress. However, NO has a short half-life and is unstable in vivo, and direct delivery to tumor sites is ineffective; therefore, increasing research focuses on NO donors. Currently, many NO donors, when combined with other antitumor drugs, exhibit higher antitumor activity than the NO donor itself or single antitumor drugs, showing promising development prospects.

[0008] Currently, single photodynamic therapy is insufficient to meet the needs of cancer treatment. Therefore, the development of highly effective combination therapies to achieve bimodal and multimodal tumor treatment has gradually become a research focus. Previous studies have shown that porphyrins and metalloporphyrin derivatives are a class of photosensitizing drugs with great potential for photodynamic therapy. They not only have a special affinity for tumors, but also produce excellent combined therapeutic effects under light conditions when combined with anticancer drugs. NO donors are also gradually demonstrating their unique advantages in tumor treatment, especially natural products containing NO donors, which can have stronger antitumor effects than the parent compound. Therefore, based on the aforementioned research on porphyrin-anticancer drugs and NO donor-photosensitizers / anticancer drugs, this invention designs and synthesizes a series of NO-type porphyrin-ferulic acid derivatives and studies their application potential in cancer treatment. Summary of the Invention

[0009] The technical problem to be solved by the present invention is to provide a NO-type porphyrin-ferulic acid derivative, which has good activity in inhibiting tumor cells.

[0010] The first aspect of the present invention is to provide a compound of formula I or formula II and its pharmaceutically acceptable salts, having the following structure:

[0011]

[0012]

[0013] Where n is an integer selected from 1 to 8;

[0014] m is selected from integers from 1 to 8;

[0015] R1 is selected from H, halogen, hydroxyl, nitro, CN, C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 6-10 Aryl, C2-C10 heteroaryl;

[0016] R is selected from H, halogen, hydroxyl, nitro, CN, C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group;

[0017] M is selected from Zn, Ni, and Mn.

[0018] Preferably, n is selected from 1, 2, 3, 4 or 5.

[0019] Preferably, m is selected from 1, 2, 3, 4 or 5.

[0020] Preferably, R1 is selected from H, halogen, hydroxyl, nitro, CN, C. 1-6Alkyl group; more preferably, R1 is selected from H or Cl;

[0021] Preferably, R is selected from H, halogen, hydroxyl, nitro, CN, C 1-6 Alkyl; more preferably, R is selected from H or methyl;

[0022] Preferably, M is selected from Zn and Ni.

[0023] Preferably, the compound structure of the present invention is as follows:

[0024]

[0025]

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

[0027]

[0028] The definitions of n, m, R1, and R are as described above.

[0029] The specific reaction steps are as follows:

[0030] In an organic solvent, a base and porphyrin derivative 5 are added, and the mixture is heated and stirred. Ferulic acid derivative 4 is then added to the reaction system until the reaction is complete. After post-treatment, compound I is obtained.

[0031] Preferably, the molar ratio of porphyrin derivative 5 to ferulic acid derivative 4 is 1:(1-1.5), more preferably 1:1-1.2, and even more preferably 1:1.2;

[0032] Preferably, the alkali is selected from potassium hydroxide, triethylamine, or potassium carbonate, and more preferably triethylamine.

[0033] Preferably, the organic solvent is selected from: dichloromethane, chloroform, tetrahydrofuran, DMSO, DMF, methanol, ethanol, isopropanol; more preferably, DMF.

[0034] Another aspect of the present invention provides a method for preparing compounds of formula II, the synthetic route of which is as follows:

[0035]

[0036] The definitions of n, m, R1, R, and M are as described above.

[0037] The specific reaction steps are as follows:

[0038] The compound of formula I was dissolved in an organic solvent, and the metal salt of M was added. The mixture was refluxed and the reaction was monitored by TLC until the reaction was complete. After post-treatment, the porphyrin-ferulic acid derivative of formula II was obtained.

[0039] Preferably, the molar ratio of compound I to metal salt M is 1:3-7, more preferably 1:5;

[0040] Preferably, the M metal salt is selected from zinc acetate dihydrate and nickel(II) acetate tetrahydrate.

[0041] Another aspect of the present invention provides a pharmaceutical composition comprising a compound of formula I or formula II or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0042] Another aspect of the present invention relates to the use of a compound of formula I or II and its pharmaceutically acceptable salts or pharmaceutical compositions comprising the same in the preparation of an anticancer drug;

[0043] Preferably, the cancer is selected from lung cancer or liver cancer; in particular, human non-small cell lung cancer cells A549 and human liver cancer cells HepG2.

[0044] Compared with the prior art, the beneficial effects of the present invention are:

[0045] (1) This invention provides a new class of NO-type porphyrin-ferulic acid derivatives with anticancer activity, which broadens the scope of existing anticancer compounds and can be used as lead compounds for further optimization.

[0046] (2) The compound of the present invention uses porphyrin molecules as carriers and utilizes their tumor tissue aggregation effect to target tumor cells, thereby reducing the side effect of killing normal cells.

[0047] (3) The porphyrin-ferulic acid compounds of the present invention can achieve synergistic treatment of phototherapy and chemotherapy. In addition, inserting metal Zn or Ni into the porphyrin can also enhance the antitumor activity of the compound.

[0048] definition:

[0049] In some embodiments, the pharmaceutically acceptable form is a pharmaceutically acceptable salt, which is well known in the art. Examples of pharmaceutically acceptable salts are those that form salts with the compound, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, perchloric acid, acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoroacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0050] "Pharmaceutically acceptable carriers" or "pharmaceutically acceptable excipients" include any and all solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic agents, and absorption-retarding agents. Pharmaceutically acceptable carriers or excipients do not impair the pharmacological activity of the disclosed compound and are non-toxic when administered in doses sufficient to deliver a therapeutic amount of the compound. The use of the aforementioned media and reagents for the pharmaceutically active substance is well known in the art. Attached Figure Description

[0051] Figure 1 shows compounds 6a-6e ( Figure 1a ), 7a-7e Figure 1b ), 8a-8e ( Figure 1c Changes in DPBF fluorescence intensity after quenching. Detailed Implementation

[0052] The present invention will be specifically illustrated below through examples. In this invention, the following examples are provided to better illustrate the invention and are not intended to limit the scope of the invention. Unless otherwise specified, the materials, reagents, etc., used in the following examples are commercially available.

[0053] Example 1: Synthesis of NO Donor 1

[0054]

[0055] At 0°C, 1.1 mL of concentrated nitric acid and 2.4 mL of concentrated sulfuric acid were slowly added to a 15 mL round-bottom flask. After stirring for 10 minutes, 1 mL of 5-bromo-1-pentanol was added, and stirring continued for 1 hour. After the reaction was complete, the reaction solution was mixed with 50 mL of ice water and extracted with CH2Cl2. The CH2Cl2 was collected, and the trace amounts of water contained in the CH2Cl2 were removed with anhydrous Na2SO4. The CH2Cl2 was then removed under reduced pressure to obtain a yellow liquid (compound 1).

[0056] 1: Yield 98%. 1 H NMR (500MHz, Chloroform-d) δ4.46 (t, J = 6.6 Hz, 2H), 3.41 (t, J = 6.7 Hz, 2H), 1.92–1.85 (m, 2H), 1.76–1.73 (m, 2H), 1.59–1.50 (m, 2H) ppm.

[0057] Example 2: Synthesis of ferulic acid derivatives 3a-3e

[0058]

[0059] 2.57 mmol of ferulic acid (500 mg) was weighed and placed in a 150 mL round-bottom flask. Thionyl chloride and DMF were added sequentially using CH₂Cl₂ as the solvent. After reacting at 40 °C for 1.5 h, 5.14 mmol of 2-bromoethanol (364.3 μL) / 3-bromo-1-propanol (464.8 μL) / 4-bromo-1-butanol (736.5 μL) / 5-bromo-1-pentanol (626.3 μL) / 6-bromo-1-hexanol (672.4 μL) was added. The reaction was monitored by thin-layer chromatography. After the reaction was complete, distilled water was added to the reaction solution to remove excess thionyl chloride and DMF. CH₂Cl₂ was collected and dehydrated with anhydrous Na₂SO₄. The crude product was obtained by rotary evaporation under reduced pressure. The crude product was eluted with CH₂Cl₂ and purified by silica gel chromatography. After collecting and concentrating the first band, a yellow, viscous liquid (compounds 3a-3e) can be obtained.

[0060] Example 3: Synthesis of ferulic acid derivatives 4a-4e containing NO donor

[0061]

[0062] Weigh 1 mmol of compounds 3a-3e (3a 301.1 mg, 3b 315.2 mg, 3c 329.2 mg, 3d 343.2 mg, 3e 357.2 mg) and 691 mg of K₂CO₃ into a 150 mL three-necked flask. Use 60 mL of acetone as solvent, and add 0.5 mmol of 5-bromonitrate and 500 μL of triethylamine. Reflux at 60 °C. After the reaction is complete, filter the reaction solution and remove acetone under reduced pressure to obtain the crude product. The crude product is also eluted with CH₂Cl₂ and purified by silica gel chromatography. Collect the second band and remove CH₂Cl₂ under reduced pressure to give a pale yellow solid, namely compounds 4a-4e.

[0063] 4a: yield 32%. 1 HNMR (500MHz, DMSO-d6) δ7.62(d,J=15.9Hz,1H),7.38(d,J=1.9Hz,1H),7.24(dd,J=8.3,1.8Hz,1H),6.98(d,J=8.4Hz,1H),6.60(d,J=15.9Hz,1H) ,4.54(t,J=6.5Hz,2H),4.47(t,J=6.5Hz,2H),4.00(t,J=6.4Hz,2H),3.8 1(s,3H),3.73(t,J=6.4Hz,2H),1.79–1.70(m,2H),1.52–1.44(m,2H)ppm.

[0064] 4b:yield 30%. 1 H NMR(500MHz,Chloroform-d)δ7.84(d,J=15.9Hz,1H),7.47(s,1H),7.29(dd,J=8.3,1.6Hz,1H),7.06(d,J=8.3Hz,1H),6.51(d,J=15.9Hz,1H),4.69(t,J=6.6Hz,2H),4.55(t,J=6.0Hz,2H),4.27(t,J=6.4Hz,2H),3.82(s,3H),3.73(t,J=6.6Hz,2H),2.42–2.52(m,2H),2.07–2.12(m,2H),1.99–2.05(m,2H),1.79–1.87(m,2H)ppm.

[0065] 4c:yield 31%. 1 H NMR(500MHz,Chloroform-d)δ7.62(d,J=15.9Hz,1H),7.08(dd,J=8.3,1.7Hz,1H),7.05(d,J=1.7Hz,1H),6.85(d,J=8.3Hz,1H),6.30(d,J=15.9Hz,1H),4.48(t,J=6.6Hz,2H),4.23(t,J=6.3Hz,2H),4.06(t,J=6.4Hz,2H),3.84(s,3H),3.47(t,J=6.6Hz,2H),2.02–1.97(m,2H),1.93–1.86(m,4H),1.84–1.79(m,2H),1.66–1.60(m,2H)ppm.

[0066] 4d:yield 33%. 1 H NMR(500MHz,Chloroform-d)δ7.62(d,J=15.9Hz,1H),7.08(dd,J=8.3,1.8Hz,1H),7.05(d,J=1.8Hz,1H),6.85(d,J=8.3Hz,1H),6.30(d,J=15.9Hz,1H),4.48(t,J=6.6Hz,2H),4.21(t,J=6.5Hz,2H),4.06(t,J=6.4Hz,2H),3.89(s,3H),3.44(t,J=6.7Hz,2H),1.93–1.87(m,4H),1.84–1.80(m,2H),1.76–1.72(m,2H),1.61–1.63(m,2H)ppm.

[0067] 4e: yield 30%. 1 H NMR(500MHz,Chloroform-d)δ7.62(d,J=15.9Hz,1H),7.08(dd,J=8.3,1.8Hz,1H),7.05(d,J= 1.8Hz,1H),6.85(d,J=8.3Hz,1H),6.30(d,J=15.9Hz,1H),4.48(t,J=6.6Hz,2H),4.20(t,J=6 .6Hz,2H),4.06(t,J=6.4Hz,2H),3.82(s,3H),3.42(t,J=6.8Hz,2H),1.91–1.87(m,4H),1.83 –1.80(m,2H),1.74–1.70(m,2H),1.65–1.58(m,2H),1.53–1.48(m,2H),1.47–1.42(m,2H)ppm.

[0068] Example 4: Synthesis of Porphyrin Matrix 5

[0069]

[0070] 4.8848 g (40 mmol) of p-hydroxybenzaldehyde and 12.7344 g (120 mmol) of benzaldehyde were weighed and placed in a 250 mL three-necked flask. The reaction was carried out at 130 °C using 150 mL of propionic acid as solvent. 11.08 mL (160 mmol) of freshly distilled pyrrole was added to a constant-pressure dropping funnel at a rate of 1 drop / second, with the titration completed within 30 min. The reaction was then stopped by reflux for 90 min. Approximately half of the propionic acid was removed by rotary evaporation, and the mixture was then placed in a refrigerator at 4 °C after adding an equal volume of ethanol. The reaction solution was filtered the next day and washed several times with ethanol to obtain a crude product with a blue-purple color. The crude product was purified by silica gel chromatography to remove impurities, yielding the target product.

[0071] 5: Yield 8%. 1 H NMR (500MHz, DMSO-d6) δ8.89–8.85(m,8H),8.22(d,J=7.9Hz,6H),8.06(d,J=8.3Hz,2H),7.80–7.73(m,9H),7.16(d,J=8.3Hz,2H),-2.76(s,2H).

[0072] Example 5 Synthesis of porphyrin-ferulic acid (6a-6e) derivatives

[0073]

[0074] Weigh 0.22 mmol of compounds 4a-4e (4a 95.01 mg, 4b 98.18 mg, 4c 101.27 mg, 4d 104.36 mg, 4e 107.44 mg), 138 mg of K₂CO₃, and 126 mg of compound 5 into a 150 mL three-necked flask. Add 200 μL of triethylamine to 60 mL of DMF as solvent and react at 60 °C. Monitor the reaction progress using thin-layer chromatography. After the reaction is complete, filter the mixture, add distilled water to the filtrate, and extract with CH₂Cl₂. Remove the water from the collected CH₂Cl₂ with anhydrous Na₂SO₄, and remove the CH₂Cl₂ under reduced pressure to obtain the crude product. Purify the crude product using silica gel column chromatography with CH₂Cl₂ as the eluent. Collect the third band to obtain the purple solid (compounds 6a-6e).

[0075] 6a: yield 80%; purity 98%; 1 H NMR(500MHz,Chloroform-d)δ8.88–8.84(m,8H),8.22(d,J=6.7Hz,6H),8.08(d,J=8.3Hz,2H),7.78–7.7 4(m,9H),7.67(d,J=15.9Hz,1H),7.21(d,J=8.3Hz,2H),7.11-7.06(m,2H),6.85(d,J=8.3Hz,1H),6.34(d ,J=15.9Hz,1H),4.52(t,J=6.1Hz,2H),4.49(t,J=6.6Hz,2H),4.06(t,J=6.4Hz,2H),3.90(s,3H),3.59(t ,J=6.1Hz,2H),1.93–1.87(m,2H),1.84–1.79(m,2H),1.65–1.62(m,2H),-2.78(s,2H)ppm; FT-IR(KBr,cm -1 ):3315(υ NH ),1705(υ C＝O ),1277(υ Ph-O-C ),1259(υ C-O-C ),1007(υ pyrrole ); HRMS-ESI:m / z calcd for C 61 H 51 N5O8 + 982.3738[M+H] + ,found 982.256.

[0076] 6b: yield 83%; purity 95%; 1H NMR(500MHz,Chloroform-d)δ8.89–8.84(m,8H),8.22(d,J=6.7Hz,6H),8.13(d,J=8.3Hz,2H),7.80–7.70(m,10H),7.30(d,J=8.4Hz,2H),7.13–7.09(m,2H),6.84(d,J=8.2Hz,1H),6.41(d,J=15.9Hz,1H),4.58(t,J=6.2Hz,2H),4.47(t,J=6.6Hz,2H),4.41(t,J=6.0Hz,2H),4.03(t,J=6.5Hz,2H),3.89(s,3H),2.43–2.38(m,2H),1.91–1.86(m,2H),1.83–1.77(m,2H),1.63–1.55(m,2H),-2.76(s,2H)ppm;FT-IR(KBr,cm -1 ):3315(υ NH ),1706(υ C＝O ),1278(υ Ph-O-C ),1254(υ C-O-C ),1001(υ pyrrole ).HRMS-ESI:m / z calcd for C 62 H 53 N5O8 + 997.3894[M+H] + ,found 997.4062.

[0077] 6c:yield 82%;purity 96%; 1 H NMR(500MHz,Chloroform-d)δ8.89–8.83(m,8H),8.22(d,J=6.3Hz,6H),8.12(d,J=8.5Hz,2H),7.80–7.74(m,9H),7.69(d,J=15.9Hz,1H),7.29(d,J=8.5Hz,2H),7.10–7.07(m,2H),6.72(d,J=8.1Hz,1H),6.38(d,J=15.9Hz,1H),4.43–4.39(m,4H),4.33(t,J=5.7Hz,2H),3.90–3.87(m,5H),2.16–2.08(m,4H),1.80–1.69(m,4H),1.52–1.47(m,2H),-2.78(s,2H)ppm;FT-IR(KBr,cm -1 ):3315(υ NH),1706(υ C＝O ),1278(υ Ph-O-C ),1252(υ C-O-C ),1001(υ pyrrole );HRMS-ESI:m / z calcd for C 63 H 55 N5O8 + 1010.4061[M+H] + ,found 1010.4151.

[0078] 6d:yield 82%;purity 96%; 1 H NMR(500MHz,Chloroform-d)δ8.89–8.83(m,8H),8.22(d,J=6.3Hz,6H),8.11(d,J=8.4Hz,2H),7.80–7.74(m,9H),7.67(d,J=15.9Hz,1H),7.28(d,J=8.5Hz,2H),7.10–7.07(m,2H),6.78(d,J=8.2Hz,1H),6.37(d,J=15.9Hz,1H),4.43(t,J=6.6Hz,2H),4.34(t,J=6.5Hz,2H),4.29(t,J=6.2Hz,2H),3.96(t,J=6.4Hz,2H),3.88(s,3H),2.09–2.03(m,2H),1.95–1.90(m,4H),1.85–1.73(m,6H),1.55–1.50(m,2H),-2.78(s,2H)ppm;FT-IR(KBr,cm -1 ):3314(υ NH ),1705(υ C＝O ),1278(υ Ph-O-C ),1256(υ c-o-c ),1001(υ pyrrole );HRMS-ESI:m / z calcd for C 64 H 57 N5O8 + 1024.4207[M+H] + ,found1024.4316.

[0079] 6e:yield 84%;purity 90%; 1H NMR(500MHz,Chloroform-d)δ8.89–8.83(m,8H),8.22(d,J=6.5Hz,6H),8.10(d,J=8.3Hz,2H),7.80–7.73(m, 9H),7.66(d,J=15.9Hz,1H),7.27(d,J=8.6Hz,2H),7.08–7.05(m,2H),6.70(d,J=8.1Hz,1H),6.36(d,J=15.9H z,1H),4.34(t,J=6.6Hz,2H),4.30(t,J=6.6Hz,2H),4.27(t,J=6.4Hz,2H),3.86(s,3H),3.81(t,J=6.4Hz,2H) ,2.04–2.00(m,2H),1.88–1.83(m,2H),1.74–1.61(m,8H),1.44–1.39(m,2H),-2.77(s,2H)ppm; FT-IR(KBr,cm -1 ):3315(υ NH ),1705(υ C＝O ),1278(υ Ph-O-C ),1254(υ C-O-C ),1001(υ pyrrole ); HRMS-ESI:m / z calcd for C 65 H 59 N5O8 + 1038.4364 [M+H] + ,found1038.4528.

[0080] Example 6 Synthesis of porphyrin-ferulic acid (7a-7e) derivatives

[0081]

[0082] Compounds 6a-6e were dissolved in CH₂Cl₂ in a 150 mL three-necked flask, and five equivalents of zinc acetate dihydrate were added. The reaction was carried out at 40 °C. After the reaction was stopped, the reaction solution and distilled water were added to a separatory funnel. The funnel was shaken to remove excess metal salts from the reaction solution by passing distilled water through it. The mixture was allowed to stand, and the organic layer was collected. The collected organic layer was dehydrated with anhydrous Na₂SO₄, and the solvent was removed under reduced pressure to obtain bright purple crystals (compounds 7a-7e).

[0083] 7a: yield 98%; purity 96%; 1H NMR(500 MHz,Chloroform-d)δ8.99–8.94(m,8H),8.22(d,J=6.8 Hz,6H),8.08(d,J=8.0 Hz,2H),7.79–7.73(m,9H),7.64(d,J=15.9Hz,1H),7.20(d,J=8.1 Hz,2H),7.08–7.05(m,2H),6.83(d,J=8.3 Hz,1H),6.31(d,J=15.9 Hz,1H),4.51–4.47(m,4H),4.05(t,J=6.4 Hz,2H),3.89(s,3H),3.58(t,J=6.1 Hz,2H),1.93–1.87(m,2H),1.85–1.79(m,2H),1.64–1.60(m,2H)ppm.FT-IR(KBr,cm -1 ):1705(υ C＝O ),1277(υ Ph-O-C ),1259(υ C-O-C ),1002(υ pyrrole );HRMS-ESI:m / z calcd for C 61 H 49 N5O8Zn + 1044.2873[M+H] + ,found 1044.1676.

[0084] 7b:yield 98%;purity 95%; 1 H NMR(500 MHz,Chloroform-d)δ9.03–8.91(m,8H),8.22(d,J=6.9 Hz,6H),8.13(d,J=8.3 Hz,2H),7.79–7.73(m,9H),7.63(d,J=15.9Hz,1H),7.29(d,J=8.3 Hz,2H),7.10–7.06(m,2H),6.82(d,J=8.2 Hz,1H),6.35(d,J=15.9 Hz,1H),4.52–4.46(m,4H),4.39(t,J=6.0 Hz,2H),4.03(t,J=6.4 Hz,2H),3.88(s,3H),2.37(t,J=6.2 Hz,2H),1.91–1.87(m,2H),1.84–1.78(m,2H),1.63–1.57(m,2H)ppm.FT-IR(KBr,cm -1 ):1707(υ C＝O ),1277(υPh-O-C ),1257(υ C-O-C ),1002(υ pyrrole );HRMS-ESI:m / z calcd forC 62 H 51 N5O8Zn + 1059.3029[M+H] + ,found 1059.3103.

[0085] 7c:yield 97%;purity 96%; 1 H NMR(500 MHz,Chloroform-d)δ8.99–8.94(m,8H),8.22(d,J=6.5 Hz,6H),8.12(d,J=8.4 Hz,2H),7.80–7.73(m,9H),7.61(d,J=15.9Hz,1H),7.28(d,J=8.4 Hz,2H),7.07–7.03(m,2H),6.72(d,J=8.2 Hz,1H),6.32(d,J=15.9 Hz,1H),4.43(t,J=6.6 Hz,2H),4.36(t,J=6.0 Hz,2H),4.31(t,J=5.8 Hz,2H),3.91(t,J=6.4 Hz,2H),3.85(s,3H),2.13–2.04(m,4H),1.83–1.72(m,4H),1.56–1.53(m,2H)ppm.FT-IR(KBr,cm -1 ):1704(υ C＝O ),1278(υ ph-O-C ),1257(υ C-O-C ),1002(υ pyrrole );HRMS-ESI:m / z calcd forC 63 H 53 N5O8Zn + 1072.3186[M+H] + ,found 1072.3384.

[0086] 7d:yield 96%;purity 94%; 1H NMR(500 MHz,Chloroform-d)δ8.99–8.94(m,8H),8.22(d,J=6.6 Hz,6H),8.11(d,J=8.3 Hz,2H),7.79–7.73(m,9H),7.61(d,J=15.9Hz,1H),7.28(d,J=8.4 Hz,2H),7.07–7.03(m,2H),6.76(d,J=8.3 Hz,1H),6.32(d,J=15.9 Hz,1H),4.45(t,J=6.6Hz,2H),4.30–4.27(m,4H),3.96(t,J=6.4Hz,2H),3.86(s,3H),2.08–2.02(m,2H),1.94–1.89(m,2H),1.85–1.74(m,6H),1.58–1.54(m,2H)ppm;FT-IR(KBr,cm -1 ):1705(υ C＝O ),1278(υ Ph-O-C ),1259(υ C-O-C ),1002(υ pyrrole );HRMS-ESI:m / z calcdfor C 64 H 55 N5O8Zn + 1086.3342[M+H] + ,found 1086.3433.

[0087] 7e:yield 98%;purity 97%; 1 H NMR(500MHz,Chloroform-d)δ8.99–8.94(m,8H),8.22(d,J=6.6Hz,6H),8.10(d,J=8.3Hz,2H),7.79–7.73(m,9H),7.60(d,J=15.9Hz,1H),7.27(d,J=7.8Hz,2H),7.05–7.01(m,2H),6.69(d,J=8.3Hz,1H),6.31(d,J=15.9Hz,1H),4.39(t,J=6.6Hz,2H),4.28–4.24(m,4H),3.85–3.83(m,5H),2.05–1.99(m,2H),1.86–1.81(m,2H),1.76–1.67(m,6H),1.64–1.59(m,2H),1.48–1.42(m,2H)ppm.FT-IR(KBr,cm -1 ):1705(υ C＝O ),1278(υPh-O-C ),1256(υ C-O-C ),1002(υ pyrrole ); HRMS-ESI:m / z calcd forC 65 H 57 N5O8Zn + 1100.3499[M+H] + ,found 1100.3702.

[0088] Example 7 Synthesis of porphyrin-ferulic acid (8a-8e) derivatives

[0089]

[0090] Compounds 6a-6e were dissolved in N,N-dimethylformamide in a 150 mL three-necked flask, and five equivalents of nickel(II) acetate tetrahydrate were added. The mixture was refluxed at 125 °C. After the reaction was complete, distilled water was added to the reaction solution, and the mixture was extracted with CH2Cl2. The CH2Cl2 was collected and a small amount of water was removed with anhydrous Na2SO4. After removing CH2Cl2 by rotation, the crude product was obtained. The crude product was purified by silica gel column chromatography using CH2Cl2 as the developing solvent to obtain reddish-brown crystals (compounds 8a-8e).

[0091] 8a: yield 52%; purity 86.93%; 1 H NMR(500MHz,Chloroform-d)δ8.78–8.74(m,8H),8.01(d,J=6.8Hz,6H),7.94(d,J=8. 2Hz,2H),7.87(d,J=8.2Hz,2H),7.74–7.62(m,10H),7.15–7.10(m,2H),6.82(d,J=7.8 Hz,1H),6.45(d,J=16.0 Hz,1H),4.71(t,J=4.7Hz,2H),4.51–4.47(m,4H),4.03(t,J=6.3 Hz,2H),3.89(s,3H),2.06–2.01(m,2H),1.93–1.78(m,4H)ppm.FT-IR(KBr,cm -1 ):1707(υ C＝O ),1277(υ Ph-O-C ),1259(υ C-O-C ),1006(υ pyrrole ); HRMS-ESI:m / z calcd for C 61 H 49 N5O8Ni +1038.2935[M+H] + ,found 1038.1083.

[0092] 8b:yield 60%;purity 82.06%; 1 H NMR(500 MHz,Chloroform-d)δ8.78–8.73(m,8H),8.01(d,J=6.8 Hz,6H),7.92(d,J=8.3 Hz,2H),7.71–7.67(m,10H),7.22(d,J=8.4 Hz,2H),7.11–7.08(m,2H),6.84(d,J=8.2 Hz,1H),6.38(d,J=15.9 Hz,1H),4.55(t,J=6.2 Hz,2H),4.35(t,J=6.0 Hz,2H),4.20(t,J=6.5 Hz,2H),4.04(t,J=6.7 Hz,2H),3.89(s,3H),2.39–2.34(m,2H),1.91–1.86(m,2H),1.78–1.72(m,2H),1.64–1.62(m,2H)ppm;FT-IR(KBr,cm -1 ):1707(υ C＝O ),1277(υ Ph-O-C ),1261(υ C-O-C ),1006(υ pyrrole );HRMS-ESI:m / z calcd forC 62 H 51 N5O8Ni + 1052.3091[M+H] + ,found 1052.1259.

[0093] 8c:yield 54%;purity 84.42%; 1H NMR(500 MHz,Chloroform-d)δ8.78–8.74(m,8H),8.01(d,J=6.8 Hz,6H),7.91(d,J=8.6 Hz,2H),7.72–7.65(m,10H),7.21(d,J=8.2 Hz,2H),7.09–7.06(m,2H),6.74(d,J=9.0 Hz,1H),6.36(d,J=15.9 Hz,1H),4.45–4.38(m,4H),4.27(t,J=7.5 Hz,2H),3.95–3.88(m,5H),2.11–2.03(m,4H),1.99–1.94(m,2H),1.82–1.73(m,4H)ppm.FT-IR(KBr,cm -1 ):1715(υ C＝O ),1277(υ Ph-O-C ),1261(υ C-O-C ),1006(υ pyrrole );HRMS-ESI:m / z calcd for C 63 H 53 N5O8Ni + 1066.3248[M+H] + ,found 1066.3314.

[0094] 8d:yield 58%;purity 85.83%. 1 H NMR(500 MHz,Chloroform-d)δ8.78–8.73(m,8H),8.01(d,J=6.7 Hz,6H),7.90(d,J=8.2 Hz,2H),7.72–7.64(m,10H),7.20(d,J=8.4 Hz,2H),7.09–7.06(m,2H),6.79(d,J=8.1 Hz,1H),6.35(d,J=15.9 Hz,1H),4.45(t,J=6.6 Hz,2H),4.32(t,J=6.4 Hz,2H),4.23(t,J=6.3 Hz,2H),3.98(t,J=6.4 Hz,2H),3.88(s,3H),2.05–1.99(m,4H),1.91–1.84(m,4H),1.79–1.74(m,4H)ppm.FT-IR(KBr,cm -1 ):1705(υ C＝O ),1276(υ Ph-O-C ),1260(υ C-O-C ),1006(υ pyrrole); HRMS-ESI:m / z calcd forC 64 H 55 N5O8Ni + 1080.3404[M+H] + ,found 1080.3488.

[0095] 8e: yield 56%; purity 95.32%. 1 H NMR(500MHz,Chloroform-d)δ8.79–8.74(m,8H),8.01(d,J=6.7Hz,6H),7.90(d,J=8.1Hz,2H),7 .70–7.63(m,10H),7.20(d,J=8.3Hz,2H),7.09–7.06(m,2H),6.73(d,J=8.1Hz,1H),6.34(d,J=15 .9Hz,1H),4.40(t,J=6.6Hz,2H),4.28(t,J=6.5Hz,2H),4.21(t,J=6.1Hz,2H),3.92–3.87(m,5H ),2.01–1.96(m,2H),1.85–1.81(m,2H),1.77–1.67(m,8H),1.53–1.45(m,2H)ppm.FT-IR(KBr,cm -1 ):1704(υ C＝O ),1277(υ Ph-O-C ),1257(υ C-O-C ),1006(υ pyrrole ); HRMS-ESI:m / z calcd for C 65 H 57 N5O8Ni + 1094.3561[M+H] + ,found 1094.3645.

[0096] Example 8 Singlet Oxygen Detection

[0097] Detection method: Accurately weigh compound 5, compounds 6a-8e, and DPBF, respectively, using chloroform as solvent, to prepare an 8 μmol / L test solution and a 20 μmol / L DPBF solution. Mix equal volumes of the DPBF solution and the test solution, and measure the fluorescence intensity change of DPBF before and after adding it to the test solution under the same conditions. Set the excitation wavelength to 418 nm and scan in the range of 200-800 nm at a scan rate of 1200 nm / min. No fluorescence was observed in any of the test compounds at 463 nm. During the detection process, the fluorescence changes of DPBF were recorded and plotted graphically; the results are shown in Figure 1 of the instruction manual.

[0098] The faster the DPBF fluorescence intensity decreases and the lower the signal value, the higher the singlet oxygen content. The results in the attached figure show that compounds 5 and 6a-8e significantly reduce the fluorescence intensity of DPBF within a short period, with the latter showing a greater decrease. Compounds 6a-6e and 7a-7e continuously reduce the fluorescence intensity of DPBF within a 50-second detection time, eventually lowering it to below 30. The fluorescence intensity of DPBF treated with compound 8a-8e decreases to around 100 within 10 seconds, indicating that compound 8a-8e can rapidly generate singlet oxygen under laser irradiation in a very short time. All these results indicate that NO-type porphyrin-ferulic acid derivatives (compounds 6a-8e) have a stronger singlet oxygen generation capacity than the porphyrin parent compound (compound 5), and can be considered as potential PDT therapeutic agents for cancer treatment.

[0099] Example 9: In vitro antitumor activity test

[0100] Both the HepG2 human liver cancer cells and the A549 human non-small cell lung cancer cells were obtained from the Cell Resource Center of the Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

[0101] Resuscitated A549 and HepG2 cells were placed in a cell culture incubator at 37°C and 5% CO2. When the cells reached approximately 90% of the culture flask's volume, the culture medium was aspirated, and residual dead cells and culture medium were washed away with PBS. 1 mL of trypsin-EDTA was added to the flask to digest the cells, and after 45 seconds, 2 mL of culture medium was added to stop the digestion. The cells were then pipetted to form a suspension, centrifuged, and the resulting cell pellet was prepared as a cell suspension for later use. Six 96-well plates were prepared, with two plates forming a group. Each group consisted of a light group (Group A) and a dark group (Group B). PBS was first added around the perimeter of each 96-well plate, 200 μL per well. Next, add 13 ml of culture medium to the sample loading well, take 185 μL of the above cell suspension, mix them, and seed them into a 96-well plate at a volume of 100 μL / well. Then, place the seeded 96-well plate into a cell culture device at an ambient temperature of 37°C and 5% CO2 for 24 hours of culture.

[0102] Each experimental drug was prepared into drug-containing culture media at concentrations of 128 μmol / L, 64 μmol / L, 32 μmol / L, and 16 μmol / L. Each experiment included a blank control group and a negative control group with six replicates, and a positive control group and an experimental group with three replicates. After removing the 96-well plate, the original culture medium was discarded, and culture medium (blank group), DMAC-containing medium (negative control group), 5-FU (positive control group), and drug-containing medium (experimental group) were added sequentially. The wells of groups A and B were then marked on the surface of the plate and placed back into the incubator for further incubation. 24 hours after drug addition, the experimental plate of group A was removed and irradiated with a near-infrared lamp for 10 min, while group B received no treatment. 48 h after drug addition, 5 g / L MTT solution was added to the A and B plates of the two experimental groups at a rate of 20 μl / well. After 4 h of incubation, the 96-well plate was removed, the supernatant was discarded, and DMSO solution was added at a rate of 150 μl / well. After the addition was complete, the wells were covered with aluminum foil. The 96-well plate containing DMSO was transferred to a shaker, and after 10 minutes, the absorbance (OD value) at 490 nm was measured using a microplate reader. The in vitro inhibitory rate (IC50) of the drug against cancer cells was calculated from the obtained OD value. 50 (Value). The test results are as follows:

[0103]

[0104]

[0105] As shown in the table above, this series of compounds also produced varying degrees of phototoxicity to A549 cells. The comparison reveals that most compounds exhibited higher phototoxicity to HepG2 cells, and compounds 7a-7c and 8a showed superior inhibitory effects on HepG2 cells compared to 5-fu under light conditions, with compound 7a showing the most significant effect (IC50 = 43.82 ± 2.50). Notably, compounds 7a and 8a also exhibited strong phototoxicity to A549 cells, and their cytotoxicity to A549 cells was stronger than that of 5-fu.

Claims

1. A compound of Formula I or Formula II and its pharmaceutically acceptable salts, having the following structure: in, n is an integer selected from 1 to 8; m is selected from integers from 1 to 8; R1 is selected from H, a halogen; R is selected from H, a halogen; M is selected from Zn and Ni.

2. The compounds of formula I or II according to claim 1 and their pharmaceutically acceptable salts, characterized in that: R1 is selected from H or Cl; R is selected from H.

3. The compounds of formula I or II according to claim 1 or 2, and their pharmaceutically acceptable salts, characterized in that: n is selected from 1, 2, 3, 4 or 5; m is selected from 1, 2, 3, 4 or 5.

4. The compounds of formula I or II according to claim 1 and their pharmaceutically acceptable salts, characterized in that, The structure of the compound is as follows: 。 5. A method for preparing the compound of formula I as described in claim 1, wherein the reaction route is as follows: ； in, The definitions of n, m, R1, and R are as described in claim 1; This includes the following reaction steps: In an organic solvent, a base and porphyrin derivative 5 are added, and the mixture is heated and stirred. Ferulic acid derivative 4 is then added to the reaction system until the reaction is complete. After post-treatment, compound I is obtained.

6. The preparation method according to claim 5, characterized in that: The molar ratio of porphyrin derivative 5 to ferulic acid derivative 4 is 1:(1-1.5). The base is selected from potassium hydroxide, triethylamine, or potassium carbonate; The organic solvent is selected from one or more of the following: dichloromethane, chloroform, tetrahydrofuran, DMSO, DMF, methanol, ethanol, and isopropanol.

7. A method for preparing the compound of formula II as described in claim 1, wherein the reaction route is as follows: in, The definitions of n, m, R1, R, and M are as described in claim 1; It includes the following reaction steps: The compound of formula I was dissolved in an organic solvent, and the metal salt of M was added. The mixture was refluxed and the reaction was monitored by TLC until the reaction was complete. After post-treatment, the porphyrin-ferulic acid derivative of formula II was obtained.

8. A pharmaceutical composition comprising a compound of formula I or formula II as claimed in any one of claims 1-4, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

9. Use of the compound of any one of claims 1-4 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 8 in the preparation of a medicament for treating cancer, wherein the cancer is selected from lung cancer or liver cancer.

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