Total synthesis method and application of metabolite psoraleic acid of psoralen

Abstract

The invention relates to the technical field of chemical synthesis, in particular to a total synthesis method and application of metabolite psoraleic acid of bakuchiol, and the total synthesis method comprises the following steps: adding a raw material bakuchiol a into an epoxidation reagent mCPBA to obtain an epoxidation compound b, performing epoxidation opening under the acid catalysis action of dilute sulphuric acid to obtain a dihydroxy compound c, and adding the dihydroxy compound c into a reaction kettle to obtain the bakuchiol metabolite. Dihydroxyl is oxidized through a strong oxidizing reagent sodium periodate to obtain an aldehyde group compound d, and finally the aldehyde group is oxidized through pinnick to obtain the target product psoraleic acid with the high yield of 90%. According to the invention, the metabolite psoraleic acid of the psoralen can be rapidly and massively obtained at the total yield of 60% through total synthesis for the first time, and a solid material foundation is laid for researching metabolic pathways and principles of psoralen series products, biological and pharmacological activities of psoralen derivatives and the like, so that the psoraleic acid becomes a good lead compound.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical synthesis, and in particular to a total synthesis method and application of psoralen acid, a metabolite of psoralen alcohol. Background Art

[0002] Bakuchiol is a phytoestrogen obtained from the seeds of Psoralea corylifolia. Bakuchiol has been shown to be a non-competitive inhibitor of multiple enzymes and has great research and application potential in anti-inflammatory, antibacterial, anti-tumor and drug metabolism regulation. Bakuchiol (5-20μM, 72h) was found to have selective cytotoxicity to A549 cells through MTT experiments. Bakuchiol is more effective than its similar substance Resveratrol (HY-16561) in reducing the viability of A549 cells. Their IC 50 s values ​​were 9.58μM and 33.02μM, respectively. Bakuchiol (5-20μM, 24h or 36h) can arrest the cell cycle progression of A549 cells to the S phase in a concentration-dependent manner and induce reactive oxygen species (ROS)-related cell apoptosis. Bakuchiol (0-80μM, 120min) is a non-competitive inhibitor of UDP-glucuronosyltransferase 2B7 (UGT2B7). Its K i The value is 10.7 μM, IC 50 The value is 40.9 μM. Bakuchiol (30 min) is a potent non-competitive inhibitor of human carboxylesterase 2 (hCE2) with a K i The value is 2.12 μM, IC 50 The value was 7.28μM. Bakuchiol (0.9775-3.91μg / mL, 24h) can increase the membrane permeability of T. mentagrophytes fungal conidia in a concentration-dependent manner. Bakuchiol (3.91μg / mL, 3h) can induce a 187% increase in the level of reactive oxygen species (ROS) in fungal cells.

[0003]

[0004] At present, although some progress has been made in the research on the source, physicochemical properties, and pharmacological effects of bakuchiol, the antibacterial, antioxidant, and anti-cancer mechanisms of bakuchiol are still not very clear, the research on its metabolic pathways in the body has not been carried out in depth, the problem of low oral bioavailability has not been solved, and the developed dosage form is too single. These have limited the widespread application of bakuchiol in clinical practice.

[0005] In the prior art, the natural product Bakuchiol has very excellent biological activity. After long-term feeding of mice Bakuchiol, psoralic acid (its metabolite psoralic acid) is separated and purified from their feces and identified. From the similarity of the structure and the content in feces, it can be considered that psoralic acid is the main product in the Bakuchiol metabolites. Metabolites are generally also the main source of new drugs, such as the third-generation antihistamine desloratadine found from the loratadine metabolites of the second-generation antihistamines, and metabolites are important contents of many drug efficacy material bases and mechanism of action research, but usually in metabolites, content is very low, and the difficulty of enrichment and purification is higher, so try to obtain its metabolites with a synthetic method. The present application completes the total synthesis of psoralic acid for the first time, and lays a material foundation for studying the biological activity and metabolic mechanism research of psoralic acid in the future. Summary of the invention

[0006] The purpose of the present invention is to solve the shortcomings existing in the prior art, and to propose a total synthesis method and application of psoralen acid, a metabolite of psoralen alcohol, to lay a solid material foundation for the biological and pharmacological activities of psoralen acid and provide a good lead compound.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A method for the total synthesis of psoralen acid, a metabolite of psoralen alcohol, comprises the following steps:

[0009] The raw material bakuchiol a is added with the classic epoxidation reagent mCPBA to obtain the epoxidation compound b, which is epoxidized under the acid catalysis of dilute sulfuric acid to obtain the dihydroxy compound c, which is oxidized with the strong oxidizing agent sodium periodate to obtain the aldehyde compound d, and finally the target product bakuchiol acid is obtained with a high yield of 90% by the classic pinnick oxidation of the aldehyde group; the synthetic reaction route is as follows:

[0010]

[0011] The present invention also provides an application of psoralen acid obtained by the above-mentioned total synthesis method in the preparation of a tool drug with the function and action mechanism of a lead compound, specifically: psoralen acid is an active pharmaceutical ingredient used as a tool drug with the function and action mechanism of a lead compound.

[0012] Preferably, the tool drug is in the form of pills, capsules, injections or oral liquids.

[0013] By adopting the above technical scheme: the natural product Bakuchiol is used as the starting material to fully synthesize the metabolite of Bakuchiol, Psoralic acid, for the first time. The natural product Bakuchiol has very excellent biological activity. By completing the full synthesis of Psoralic acid for the first time, a material foundation is laid for future research on the biological activity and metabolic mechanism of Psoralic acid. The invention is the first to fully synthesize Psoralic acid through 4 steps of organic reaction, with a total yield of 60.6%.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention can for the first time rapidly and in large quantities obtain the metabolite of psoralen by total synthesis with a total yield of 60%, thereby laying a solid material foundation for studying the metabolic pathways and principles of psoralen series products, the biological and pharmacological activities of psoralen derivatives, etc., making it a good lead compound. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The nuclear magnetic resonance hydrogen spectrum of the intermediate d of psoralen acid of the present invention;

[0017] Figure 2 The psoralen nuclear magnetic resonance 1 H-NMR spectrum;

[0018] Figure 3 The psoralen nuclear magnetic resonance 13 C-NMR carbon spectrum;

[0019] Figure 4 It is the HMBC two-dimensional nuclear magnetic resonance spectrum of psoralen acid of the present invention;

[0020] Figure 5 The figure is the two-dimensional nuclear magnetic resonance HSQC spectrum of psoralic acid of the present invention. DETAILED DESCRIPTION

[0021] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the accompanying drawings, so that those skilled in the art can better understand the advantages and features of the present invention, thereby making a clearer definition of the protection scope of the present invention. The embodiments described in the present invention are only a part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work are within the scope of protection of the present invention.

[0022] Reference Figure 1-Figure 5 ,in, Figure 1 The key precursor intermediate d for the total synthesis of psoralen acid of the present invention 1H-NMR spectrum, the target product of the present invention, psoralen acid, is obtained by pinnick oxidation. Figure 2 , Figure 3 , Figure 4 , Figure 5 The nuclear magnetic resonance analysis was used to characterize psoralen and confirm the structure of the compound.

[0023] Embodiment 1:

[0024]

[0025] Bakuchiol a (1.282 g, 5.0 mmol) was dissolved in 30 ml of dichloromethane solution, cooled to 0°C in an ice-water bath, and m-CPBA (0.949 g, 5.5 mmol) dissolved in 60 ml of dichloromethane was slowly added dropwise, and the reaction was allowed to proceed at room temperature for 4 hours. The reaction was monitored by TLC, and 30 ml of saturated sodium bisulfate solution was added to quench the reaction, and the reaction was stirred for 30 minutes. Extracted with dichloromethane 3 times, 30 ml each time, combined with dichloromethane, washed once with 30 ml of saturated brine, and the dichloromethane organic phase was dried by adding anhydrous magnesium sulfate. After drying for half an hour, the magnesium sulfate was removed by filtration, and dichloromethane was removed by vacuum distillation, and then petroleum ether-ethyl acetate (PE:EA=2:1) ​​column chromatography was used to obtain white solid compound b (1.158 g, yield 85%).

[0026] 1 H NMR (400MHz, CDCl3-d): δ7.27(d,J=8.5Hz,2H),6.78(d,J=8.5Hz,2H),6.30(d,J=16.2Hz,1H),6.09(d,J=16.2Hz,1H),5.9 4-5.90(m,1H),5.08-5.02(m,1H),5.05-5.00(m,1H),2.64(t,J=6.0Hz,1H),1.60-1.55(m,4H),1.22(s,3H),1.19(s,6H).

[0027] Embodiment 2:

[0028]

[0029] Epoxide b (817.2 mg, 3.0 mmol) was dissolved in 50 ml of dioxane: water = 1: 1 solution (25 ml of dioxane and 25 ml of water), and 5 drops of concentrated sulfuric acid were added with a rubber dropper. The reaction was allowed to react overnight at room temperature. The reaction raw material spot disappeared after monitoring by TLC. The dioxane was removed by vacuum distillation. The remaining aqueous phase was extracted with a mixed solution of dichloromethane: isopropanol = 3: 1 for 4 times, 30 ml each time, and the organic layers were combined. The dichloromethane organic phase was dried by adding anhydrous magnesium sulfate. After drying for half an hour, the magnesium sulfate was removed by filtration. After dichloromethane was removed by vacuum distillation, the dihydroxy compound c was obtained by column chromatography using petroleum ether-ethyl acetate (PE: EA = 1: 1), 862.5 mg (yield 99%).

[0030] 1 H NMR (400MHz, CDCl3-d): δ7.20(d,J=8.5Hz,2H),6.72(d,J=8.5Hz,2H),6.23(d,J=1 5.9Hz,1H),6.05(d,J=15.9Hz,1H),5.86(dd,J=10.8,17.6Hz,1H),4.96(d,J=17.6 Hz,1H),4.95(d,J=10.8Hz,1H),3.18(dd,J=5.0,12.0Hz,1H),1.56-1.52(m,1H),1 .88-1.85(m,1H),1.42-1.38(m,1H),1.25-1.21(m,1H),1.05(s,6H),1.13(s,3H);.

[0031] Embodiment 3:

[0032]

[0033] Dissolve the dihydroxy compound c (1.162g, 4.0mmol) in 50ml tetrahydrofuran: water = 3:2 solution (30ml tetrahydrofuran and 20ml water), add sodium periodate (1.711g, 8.0mmol), react at room temperature for 1 hour, TLC monitoring reaction raw material point disappears, remove tetrahydrofuran by vacuum distillation, extract the remaining aqueous phase with dichloromethane: isopropanol = 3:1 mixed solution 4 times, 30ml each time, and combine the organic layers. Add anhydrous magnesium sulfate to the dichloromethane organic phase and dry it. After drying for half an hour, filter to remove magnesium sulfate, remove dichloromethane by vacuum distillation, and use petroleum ether-ethyl acetate (PE:EA = 3:1) column chromatography to obtain aldehyde compound d, (781.8mg, yield 80%).

[0034] 1H NMR (400MHz, CDCl3-d): δ7.22(d,J=6.4Hz,2H),6.76(d,J=8.5Hz,2H),6.25(d,J=16.2Hz,1H),5.98(d,J=16.2Hz ,1H),5.86(d,J=10.7Hz,1H),5.06(s,1H),2.42–2.24(m,2H),1.95–1.72(m,2H),1.34-1.25(m,3H),1.18(s,3H).

[0035] Embodiment 4:

[0036]

[0037] The aldehyde compound d (244.3 mg, 1.0 mmol) was dissolved in 20 ml of tert-butanol, and 15 ml of aqueous solution prepared by sodium dihydrogen phosphate (960.0 mg, 8.0 mmol) and sodium hypochlorite (670.0 mg, 9.0 mmol) was added, and the auxiliary agent 2-methyl-2-butene (2.800 g, 4.25 ml, 40.0 mmol) was added. The reaction was carried out at room temperature for 2 hours. After TLC monitoring, the reaction raw material point disappeared, and the tert-butanol was removed by vacuum distillation. The remaining aqueous phase was extracted with a mixed solution of dichloromethane: isopropanol = 3:1 for 4 times, 30 ml each time, and the organic layers were combined. The dichloromethane organic phase was dried by adding anhydrous magnesium sulfate, and the magnesium sulfate was removed by filtration after drying for half an hour. After dichloromethane was removed by vacuum distillation, petroleum ether-ethyl acetate (PE:EA = 1:5) was used for column chromatography to obtain psoralen acid, 234.3 mg (yield 90%).

[0038] 1 H NMR (400MHz, CDCl3-d): δ7.22(d,J=8.3Hz,2H),6.80–6.72(m,2H),6.25(d,J=16.2Hz,1H),5.98(d,J=16.3Hz,1H),5. 84(dd,J=17.4,10.7Hz,1H),5.09–5.01(m,2H),2.37–2.28(m,2H),1.88–1.80(m,2H),1.34–1.25(m,3H),1.18(s,3H).

[0039] In summary, the present invention can for the first time obtain the metabolite of psoralen, psoralen acid, quickly and in large quantities with a total yield of 60% through total synthesis, laying a solid material foundation for studying the metabolic pathways and principles of psoralen series products, the biological and pharmacological activities of psoralen derivatives, etc., making it a good lead compound.

[0040] The description and practice disclosed in the present invention are easy to think and understand for ordinary technicians in the technical field, and several improvements and modifications can be made without departing from the principles of the present invention. Therefore, modifications or improvements made without departing from the spirit of the present invention should also be regarded as the protection scope of the present invention.

Claims

1. A method for the total synthesis of psoralen acid, a metabolite of psoralen alcohol, characterized in that: The steps include: The raw material bakuchiol a is added with the epoxidation reagent mCPBA to obtain the epoxidation compound b, which is epoxidized under the acid catalysis of dilute sulfuric acid to obtain the dihydroxy compound c, which is oxidized with the strong oxidizing reagent sodium periodate to obtain the aldehyde compound d, and finally the target product bakuchiol acid is obtained with a high yield of 90% by pinnick oxidation of the aldehyde group. The synthetic reaction route is as follows:

2. An application of psoralen acid obtained by the total synthesis method of claim 1 in the preparation of tool drugs with the function and mechanism of action of lead compounds.

3. The use according to claim 2, characterized in that: The tool medicine is in the form of pills, capsules, injections or oral liquids.

Images