Ferroptosis-Inducing Nanoparticle Formulation Targeting Ferritin, Preparation Method Thereof, and Application

Abstract

The present invention designs a class of ferroptosis-inducing nanodrugs targeting ferritin based on PROTAC design, and its general structural formula is shown in I: #imgabs0# Among them, #imgabs1# represents a ligand of E3 ubiquitin ligase, -L- represents a linking group, which is selected from an unsubstituted or optionally C-containing one or more heteroatoms 1‑50 alkyl chain, and the heteroatoms can be selected from O, S, N; #imgabs2# represents various saturated or unsaturated long alkyl chains. For ferritin for which there is no commercial inhibitor, it can degrade the target protein in cells at a lower concentration, synergistically induce iron ion overload with iron-based metal-organic frameworks, induce the Fenton reaction to generate reactive oxygen species (ROS), thereby leading to the accumulation of lipid peroxidation and inducing ferroptosis of cells. This class of compounds has been further verified in pancreatic cancer, has good degradation activity and tumor killing effect, and has high safety and broad application prospects.

Description

Technical Field:

[0001] The present invention relates to the field of biomedical technologies, and in particular to a ferroptosis-inducing nanodose form targeting ferritin, a preparation method thereof, and an application. Background Art:

[0002] Cancer is one of the important diseases affecting human health and has now become the second leading cause of death globally. Cancer treatment mainly includes traditional surgical treatment, radiotherapy, chemotherapy, and new treatment methods such as targeted therapy and immunotherapy that have developed rapidly in recent years. Due to the continuous replenishment of growth-promoting signals in tumors; extensive inhibition of anti-growth signals; resistance to cell death; unlimited replication ability; continuous angiogenesis; tissue infiltration and metastasis; avoidance of immune destruction; promotion of tumor inflammation; abnormal cell energy metabolism; genomic instability and mutations, etc., it has caused great obstacles to the treatment of the disease. Therefore, a single treatment method is difficult to achieve therapeutic benefits. For the treatment of cancer, a combination of multiple methods is often used, such as the combined application of surgical resection, radiotherapy, and chemotherapy. Among them, due to the rapid progression of cancer diseases, the best periods for surgical treatment and radiotherapy are often missed at the time of diagnosis, and chemotherapy drugs are needed to extend the survival of patients. Therefore, there is an urgent need for some anti-tumor compounds with excellent efficacy in clinical practice. Currently, most anti-tumor drugs in clinical use exert their anti-tumor effects partially or entirely through apoptosis, such as platinum compounds, paclitaxel, irinotecan, gemcitabine, temozolomide, pemetrexed, imatinib, etc. In the long-term clinical application of these drugs, they will induce apoptosis resistance in cancer cells, making it difficult to kill tumor cells and resulting in good therapeutic benefits for patients. To address this problem, developing new drugs that induce non-apoptotic cell death is one of the strategies for killing tumor cells and overcoming apoptosis resistance. Among various forms of non-apoptotic cell death, in 2012, ferroptosis discovered by Dixon, which is mediated by lipid peroxidation, is a good anti-tumor strategy.

[0003] The main regulators of ferroptosis are iron ions, lipid peroxidation, and the thiol reduction system. Under the action of divalent iron or lipoxygenase, unsaturated fatty acids highly expressed on the cell membrane are catalyzed to undergo lipid peroxidation, thereby inducing cell death. Under normal circumstances, GPX4, the regulatory core enzyme of the antioxidant system (glutathione system), inhibits the above pathway through a feedback mechanism, and the dysregulation of GPX4 will also induce the occurrence of ferroptosis. Currently, marketed ferroptosis inducers such as Erastin and soraferib mainly act by regulating the thiol reduction system to inhibit the reduction of lipid peroxides. Drugs targeting iron ion metabolism regulation are relatively rare. In the previous research of this research group, Fe was mainly delivered through the MOF-Fe nanodose form 2+To induce ferroptosis, it was found that MOF-Fe could effectively deliver iron ions into tumors and kill some tumor cells, but while the intracellular Fe 2+ increased, the cell protection mechanism was also upregulated, resulting in the upregulation of FTH1 and the storage of the overloaded Fe 2+ Therefore, in order to release Fe from the protein, induce iron overload, and further kill tumor cells, ferritin needs to be degraded. However, due to the special structure of ferritin, there is currently no targeted inhibitor for ferritin.

[0004] The chemical biology proteolysis targeting chimeras (PROTACs) are a class of bifunctional small molecule compounds based on the ubiquitin-proteasome system that chemically induce the polyubiquitination and degradation of target proteins. The structure contains two ligands: one end is a ligand that can recruit the protein degradation system (such as E3 ubiquitin ligase), and the other end is a ligand that binds to the target protein in the cell. The two ligands are connected by a linker. After PROTACs enter the cell, the ligands at both ends specifically bind to the corresponding proteins, bringing the target protein and the intracellular E3 ubiquitin ligase closer together to form a "target protein-PROTAC-E3" ternary complex. Among them, the E3 ubiquitin ligase can mediate the ubiquitination of the target protein by ubiquitin-conjugating enzyme E2. After the ternary complex dissociates, the dissociated PROTACs can bind to the protein again, and the target protein "labeled" by ubiquitin is recognized and degraded by the proteasome, thereby selectively reducing the level of the target protein in the cell.

[0005] Ferritin is a widely existing iron storage protein, with a protein shell in the form of an inner hollow structure composed of 24 subunits in a highly symmetric manner. The hollow diameter is about 8 nm. The protein shell surrounds an iron core composed of iron and phosphate molecules, with an outer diameter of 12 - 13 nm and a molecular weight of 500 kDa. Ferritins from different sources vary greatly in their primary structure, making it difficult to design specific inhibitors. Therefore, using chemical biology PROTACs to design and synthesize ferritin has good application prospects.

[0006] The iron storage protein ferritin contains fatty acid binding sites, and its occupancy regulates the absorption and release of iron. Therefore, it can be used as a binding ligand for ferritin. In the ferroptosis metabolic pathway, monounsaturated fatty acids often inhibit the occurrence of ferroptosis, while polyunsaturated fatty acids promote ferroptosis. Selecting polyunsaturated fatty acids to synthesize a series of ferritin degradants may have better therapeutic effects.

[0007] The molecular weights of PROTACs generally range from 600 to 1300, which are relatively large among small molecules. In addition, due to the non-compliance of molecular design with the "five principles of drug-likeness", the membrane permeability and bioavailability of compounds are poor, and the drug-forming performance is poor. Therefore, in order to further improve the in vivo bioavailability and drug-forming performance of drugs, it is necessary to further improve through certain drug dosage form designs. MOF-Fe@PROTAC@PEI has good solubility, high in vivo bioavailability, and can also deliver iron ions into cells to play a synergistic sensitization role with ferritin PROTAC.

[0008] Therefore, developing drugs with efficient targeting and ferritin degradation through simple chemical means for PROTAC drug development technology will have broader application prospects and market value. Summary of the Invention:

[0009] The purpose of the present invention is to provide an iron death-induced nano-formulation targeting ferritin, its preparation method and application, so as to solve the deficiencies of the prior art.

[0010] The present invention is implemented by the following technical solutions: On the one hand, the present invention provides a compound of general formula I:

[0011]

[0012] Among them, represents a ligand of E3 ubiquitin ligase;

[0013] The -L- represents a linking group, which can be unsubstituted or an optionally substituted C 1-50 alkyl chain, and the heteroatom can be selected from O, S, N.

[0014] The represents various saturated or unsaturated long alkyl chains, which are formed by the reaction of saturated or unsaturated fatty acids and their acyl halide and anhydride derivatives with amino groups. The above-mentioned saturated or unsaturated fatty acids can be selected from palmitic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, etc.

[0015] According to the embodiments of the present invention, the can be selected from conventional E3 ubiquitin ligase ligands (those skilled in the art of pharmacy can understand that the ligand is connected to -L- at any suitable site). Examples of the ligand structure are as follows:

[0016]

[0017]

[0018]

[0019] According to a preferred embodiment of the present invention, the may have the following structure;

[0020]

[0021] According to an embodiment of the present invention, the general formula I is further selected from the following structures of formula II, formula III, and formula IV:

[0022]

[0023] In formula II, -L-, is as defined above;

[0024]

[0025] In formula III, is as defined above, and n is selected from 1 to 100;

[0026]

[0027] In formula IV, is as defined above, and n is selected from 1 to 100;

[0028] The present invention provides a method for preparing a compound of general formula I, comprising the following steps:

[0029]

[0030]

[0031] According to an embodiment of the present invention, the preparation method of the general formula I may specifically further include one or more of the following steps:

[0032] Step 1: Synthesis of c and d type molecules:

[0033] Dissolve compound a in organic reagent A, select Boc-protected amino acid molecules with different lengths (1.2 eq), add condensation reagent A, react under nitrogen protection at room temperature for 2 - 4 hours, extract the reaction crude product with 5% sodium carbonate solution and organic reagent B, and purify by silica gel column to obtain the target product c. The synthesis of molecule d is the same as that of molecule c.

[0034] Step 2: Synthesis of e and f type molecules:

[0035] Dissolve molecule c in dichloromethane, add hydrochloric acid (4M) - 1,4-dioxane (3 eq), react at room temperature for 30 minutes, and obtain the crude product e by rotary evaporation. The synthesis of molecule f is the same as that of molecule e.

[0036] Step 3. Synthesis of OALnV and OALnOV molecules:

[0037] Dissolve compound e in organic reagent A, select different fatty acids or their anhydrides and acyl halide derivatives (1 eq), add condensation reagent C (2 eq) and the accompanying base solution, react overnight under nitrogen protection at room temperature, extract the crude reaction product with 5% sodium carbonate solution and organic reagent B, and purify it by silica gel column to obtain the target product OALnV. The synthesis of molecule OALnV is the same as that of molecule OALnOV.

[0038] According to the embodiments of the present invention, for the preparation method of the compound of general formula I, the organic reagent A is selected from dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, and dichloromethane; the organic reagent B is selected from ethyl acetate and dichloromethane; the condensation reagent C is selected from O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU), N,N'-carbonyldiimidazole (CDI), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (TCFH).

[0039] According to the embodiments of the present invention, those skilled in the relevant pharmaceutical fields can understand that in the above preparation method, the final product can be synthesized starting from any intermediate.

[0040] On the other hand, the present invention provides a cationic polyimine-coated MOF-Fe nanoparticle, which internally loads the compound of general formula I described above.

[0041] According to the embodiments of the present invention, for the MOF-Fe nanoparticle, the MOF-Fe loads the compound of general formula I through its internal pore diameter, and the surface of MOF-Fe@PROTAC is coated with cationic polyimine to prepare the MOF-Fe@PROTAC@PEI complex.

[0042] According to the embodiments of the present invention, for the nanoparticle, the cationic polyimine includes one or more of polyetherimide (PEI) and polyoxyalkylene polyamine (ODA).

[0043] According to the preparation method of the above MOF-Fe nanoparticle, it includes the following steps:

[0044] (1) Add 2-aminoterephthalic acid (NH2-BDC) and iron(III) chloride hexahydrate (FeCl3·6H2O) to the reaction kettle, and react at 110 °C for 48 h.

[0045] (2) After drying in an oven at 100 °C, add PROTAC, disperse it by ultrasound, and incubate at room temperature for 24 h.

[0046] (3) The above-mentioned nanoparticles were centrifuged and washed clean, incubated with cationic polyimine for 24 h, the precipitate was collected by centrifugation and resuspended.

[0047] In another aspect, the present invention provides the use of the compound of general formula I or its nano-drug MOF-Fe@PROTAC@PEI in the preparation of anti-tumor drugs. According to some preferred embodiments of the present invention, the tumors include: human pancreatic ductal carcinoma (PANC-1, MIAPACA2) and murine cell line KPC-A719 pancreatic cancer cells (LSL-Kras G12D / + ; LSL-Trp53 R172H / + ; Pdx-1-Cre).

[0048] In another aspect, the present invention provides the use of the compound of general formula I or its nano-drug MOF-Fe@PROTAC@PEI in the preparation of ferroptosis inducers. According to the embodiments of the present invention, the diseases that the ferroptosis inducer can prevent and / or treat include neurodegenerative diseases, traumatic and hemorrhagic brain injuries, ischemic injuries (liver, kidney and heart ischemic injuries)

[0049] In some embodiments, the diseases include Parkinson's disease (PD), Huntington's disease (HD) and Alzheimer's disease (AD).

[0050] Advantages of the present invention:

[0051] 1. In the present invention, polyunsaturated fatty acids and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride which recruits E3 ubiquitin ligase are used to form Ferritin-PROTAC series structures shown in general formulas II, III and IV by matching and optimizing linkers. It has been found through research that this PROTAC can indeed cause the degradation of ferritin in different cell lines, as well as in cells with high expression and overexpression of ferritin, and further induce ferroptosis. Research shows that the combined use of Ferritin-PROTAC and MOF-Fe can play a strong synergistic effect and further enhance the tumor killing effect of Ferritin-PROTAC.

[0052] 2. The inventor team found through experimental research that the compound series of general formulas II, III, and IV have relatively high efficiency in killing pancreatic cancer cells. In particular, C20U4LV with arachidonic acid as the ferritin ligand has the highest degradation efficiency and relatively high tumor-killing effect. The ROS content on the cell membrane measured by BODIPY"581 / 591C11 increases with the increase in concentration, further confirming the occurrence of ferroptosis. By comparing different pancreatic cancer cell lines in the MTT cytotoxicity experiment, it is confirmed that C20U4LV can act on different cell lines, and it has a better effect in the cell MIAPACA2 with high ferritin expression. It also has a good protein degradation effect on the PANC-1 cell with overexpressed ferritin in the simulated drug-resistant cell line.

[0053] 3. The research team of the present invention further found that the combination of MOF-Fe and Ferritin-PROTAC has a strong tumor-killing effect and enhances the curative effect of ferroptosis. Therefore, a drug delivery strategy of MOF-Fe@PROTAC@PEI was designed. The cationic PEI was used to improve the delivery efficiency of PROTAC small molecules, increase the stability of the nano-drug, increase the water solubility of the drug, thereby further improving the targeting of this type of drug and reducing the toxic effect on normal cells. BRIEF DESCRIPTION OF THE DRAWINGS:

[0054] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0055] Figure 1 Schematically shows the structure and working principle of the optimal ferritin PROTAC degrader C18U2L3V.

[0056] Figure 2 Schematically shows the IC 50 (Cell viability) and AUC area (drug sensitivity).

[0057] Figure 3 Schematically shows the effect of the combined application of iron-based metal-organic framework (MOF-Fe) and ferritin PROTAC degrader C20U4LV on the proliferation of PANC-1 cells.

[0058] Figure 4 Schematically shows the ferritin degradation activity and statistical results of C20U4LV, C20U4L4V, C20U2L3V, and C20U3L4OV combined with MOF-Fe.

[0059] Figure 5 Schematic illustration of the degradation activity of C20U4LV in combination with MOF-Fe on ferritin in human pancreatic cancer cell line PANC-1 under the action of the ubiquitin ligase ligand (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (VHL-ligand).

[0060] Figure 6 Schematic illustration of the scanning electron microscope (SEM) image of MOF-Fe@C20U4LV@PEI.

[0061] Figure 7 1H-NMR results of C18U2L3V.

[0062] Figure 8 13C-NMR results of C18U2L3V.

[0063] Figure 9 ESI mass spectrum of C18U2L3V.

[0064] Figure 10 1H-NMR results of C18U2L4OV.

[0065] Figure 11 13C-NMR results of C18U2L4OV.

[0066] Figure 12 ESI mass spectrum of C18U2L4OV.

[0067] Figure 13 1H-NMR results of C20U4LV.

[0068] Figure 14 13C-NMR results of C20U4LV.

[0069] Figure 15 ESI mass spectrum of C20U4LV.

[0070] Figure 16 1H-NMR results of C20U4L4V.

[0071] Figure 17 13C-NMR results of C20U4L4V.

[0072] Figure 18 ESI mass spectrum of C20U4L4V.

[0073] Figure 19 1H-NMR results of C18U1L3V.

[0074] Figure 20 13C-NMR results of C18U1L3V

[0075] Figure 21 ESI mass spectrum of C18U1L3V Specific implementation method:

[0076] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0077] Unless otherwise specified, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known and mature methods.

[0078] Example 1 Synthesis of C20U4LV: (when n = 0)

[0079] Step 1 Synthesis of C20U4LV: Dissolve (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (1eq, 100mg) that recruits E3 ubiquitin ligase and arachidonic acid (1.5eq, 97mg) in the organic reagent DMF, add the condensation reagents HATU (2eq, 155mg) and DIPEA (6eq, 120mg), protect with nitrogen, react overnight at room temperature, extract the crude reaction product with 5% sodium carbonate solution and DCM, and purify by silica gel column to obtain a light yellow solid. 1 1H NMR (600 MHz, CDCl3) δ 8.69 (s, 1H), 7.38 – 7.31 (m,4H), 6.33 (d, J J = 9.0 Hz, 1H), 5.41 – 5.32 (m, 8H), 4.69 (t, J J = 7.9 Hz, 1H),4.59 – 4.51 (m, 3H), 4.33 (dd, J J = 15.0, 5.3 Hz, 1H), 4.06 (d, J J = 11.4 Hz, 1H),3.64 (dd, J= 11.2, 3.8 Hz, 1H), 2.88 – 2.76 (m, 7H), 2.51 (s, 3H), 2.49 – 2.45(m, 1H), 2.33 (t, 1H), 2.20 (t, J = 7.4 Hz, 2H), 2.16 – 2.01 (m, 5H), 1.74 –1.61 (m, 3H), 1.36 (q, J = 7.5 Hz, 2H), 1.26 (s, 1H), 0.94 (s, 9H), 0.88 (t, J =7.0 Hz, 3H). 13 C NMR (151 MHz, CDCl3) δ 176.65, 173.63, 171.87, 170.83, 150.44,148.39, 138.10, 131.63, 130.91, 130.52, 130.50, 129.51, 128.95, 128.91,128.86, 128.63, 128.59, 128.29, 128.23, 128.15, 128.11, 128.07, 127.86,127.83, 127.54, 127.51, 70.00, 58.70, 57.45, 56.77, 43.22, 36.01, 35.84,35.10, 33.44, 31.51, 29.69, 29.31, 27.22, 26.62, 26.58, 26.41, 25.64, 25.62,25.47, 24.77, 22.57, 16.00, 14.09. ESI-MS(m / z): [M+Na] calculated forC 42 H 60 N4O4S: 739.4233; found 739.420.

[0080] Example 2

[0081] Referring to the synthesis of Example 1, oleic acid was used as a raw material for replacement, and the molecule of C18U1LV was synthesized (the structure is shown below).

[0082]

[0083] 11H NMR (600 MHz, CDCl3) δ 8.66 (s, 1H), 7.33 (q, J = 8.1 Hz, 5H), 6.17 (t, J = 9.2 Hz, 1H), 5.35 – 5.28 (m, 2H), 4.69 – 4.65 (m, 1H), 4.55 – 4.48 (m, 3H), 4.33 (dd, J = 15.0, 5.4 Hz, 1H), 4.03 (d, J = 9.1 Hz, 1H), 3.73 – 3.65 (m, 2H), 3.62 (dd, J = 11.2, 3.8 Hz, 1H), 3.17 (q, J = 7.7 Hz, 2H), 2.49 (s, 3H), 2.15 (t, J = 7.7 Hz, 2H), 2.12 – 2.07 (m, 1H), 1.98 (q, J = 7.2 Hz, 4H), 1.32 – 1.22 (m, 20H), 0.92 (s, 9H), 0.86 (t, J = 6.9 Hz, 3H).

[0084] 13 13C NMR (151 MHz, CDCl3) δ 173.79, 171.79, 170.88, 150.35, 148.44, 138.11, 131.60, 130.93, 130.03, 129.70, 129.50, 128.07, 70.02, 58.70, 57.45, 56.69, 55.81, 55.73, 43.64, 43.20, 36.55, 31.90, 29.76, 29.70, 29.52, 29.32, 29.23, 29.20, 29.14, 27.22, 27.17, 26.42, 25.67, 22.68, 18.53, 18.19, 17.10, 16.67, 16.05, 14.13, 12.62. ESI-MS(m / z): [M+Na] calculated for C 40 H 62 N4O4S: 717.4384; found 717.437

[0085] Example 3

[0086] Referring to the synthesis of Example 1, oleic acid was used as a raw material for replacement, and the molecule of C18U3LV was synthesized (the structure is shown below).

[0087]

[0088] 1 1H NMR (600 MHz, CDCl3) δ 8.66 (s, 1H), 7.33 (p, J J = 8.6 Hz, 5 H), 6.15(d, J J = 8.7 Hz, 1H), 5.41 – 5.25 (m, 6H), 4.69 (t, J J = 7.9 Hz, 1H), 4.58 – 4.47(m, 3H), 4.32 (dd, J J = 15.0, 5.3 Hz, 1H), 4.06 (d, J J = 11.4 Hz, 1H), 3.71 – 3.67(m, 4H), 3.60 (dd, J J = 11.3, 3.7 Hz, 1H), 2.50 (s, 3H), 2.17 (t, J J = 7.6 Hz, 2H),2.08 – 2.00 (m, 5H), 1.60 – 1.55 (m, 2H), 1.28 – 1.24 (m, 7H), 0.95 (t, J J = 7.5Hz, 3H), 0.91 (s, 9H). 13 13C NMR (151 MHz, CDCl3) δ 173.79, 171.95, 170.74,162.70, 150.34, 148.44, 138.10, 131.97, 131.62, 130.95, 130.25, 129.53,128.30, 128.24, 128.13, 127.74, 127.11, 70.04, 58.52, 57.45, 56.63, 55.45,43.45, 43.25, 36.58, 36.52, 35.85, 34.92, 31.48, 29.58, 29.22, 29.19, 29.13,27.20, 26.42, 25.62, 25.53, 20.55, 18.55, 17.10, 16.05, 14.29, 12.67. ESI-MS(m / z): [M+Na] calculated for C 40 H 58 14N4O4S: 713.4071; found 713.402

[0089] Example 4

[0090] Referring to the synthesis of Example 1, cis-5,8,11,14,17-eicosapentaenoic acid was used as a raw material for replacement, and the molecule of C20U5LV was synthesized (the structure is shown below).

[0091]

[0092] 1 H NMR (600 MHz, CDCl3) δ 8.67 (s, 1H), 8.00 (s, 1H), 7.37 – 7.32 (m,4H), 7.29 (t, J = 6.0 Hz, 1H), 6.15 (d, J = 8.7 Hz, 1H), 5.42 – 5.26 (m, 10H),4.70 (t, J = 7.9 Hz, 1H), 4.59 – 4.48 (m, 3H), 4.32 (dd, J = 14.9, 5.2 Hz, 1H),4.08 (d, J = 11.5 Hz, 1H), 3.75 – 3.67 (m, 2H), 3.60 (dd, J = 11.3, 3.7 Hz, 1H),3.21 – 3.14 (m, 2H), 2.50 (s, 3H), 2.19 (t, J = 7.7 Hz, 2H), 2.11 – 2.02 (m,4H), 1.66 (dd, J = 13.6, 7.5 Hz, 2H), 1.41 (d, J = 6.6 Hz, 6H), 0.96 (t, J = 7.5Hz, 3H), 0.92 (s, 8H). 1313C NMR (151 MHz, CDCl3) δ 173.61, 171.93, 170.68, 162.68, 150.35, 148.45, 138.06, 132.06, 131.61, 130.98, 129.54, 128.89, 128.60, 128.31, 128.20, 128.13, 128.07, 127.85, 127.00, 70.06, 58.49, 57.50, 56.66, 55.63, 43.59, 43.27, 36.56, 35.84, 35.79, 34.87, 31.48, 26.62, 26.41, 25.63, 25.55, 25.43, 20.56, 18.58, 17.16, 16.04, 14.28, 12.59. ESI-MS (m / z): [M+Na] calculated for C 42 H 58 N4O4S: 737.4071; found 737.404.

[0093] Example 5 Synthesis of C20U4L1V: (when n = 1)

[0094]

[0095] Step 1. Synthesis of Compound 1: Dissolve (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (1 eq, 100 mg) which recruits E3 ubiquitin ligase and arachidonic acid (1.5 eq, 97 mg) in the organic reagent DMF. Add the condensation reagents HATU (2 eq, 155 mg) and DIPEA (6 eq, 120 mg), protect with nitrogen, react overnight at room temperature, extract the crude reaction product with 5% sodium carbonate solution and DCM, and purify by silica gel column to obtain a pale yellow solid.

[0096] Step 2. Synthesis of Compound 2:

[0097] Dissolve Molecule 1 in dichloromethane, add hydrochloric acid (4M)-1,4-dioxane (3 eq), react at room temperature for 30 minutes, and obtain the crude product 2 after rotary evaporation.

[0098] Step 3. Synthesis of Compound C20U4L1V:

[0099] Compound 2 and arachidonic acid (1.5 eq, 74.3 mg) were dissolved in the organic reagent acetonitrile. The condensation reagents HATU (2 eq, 155 mg) and DIPEA (6 eq, 150 mg) were added. Under nitrogen protection, the reaction was carried out overnight at room temperature. The crude reaction product was obtained by extraction with 5% sodium carbonate solution and DCM, and purified by silica gel column chromatography to obtain a white solid.

[0100] 1 H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 8.01 (t, J = 6.0 Hz,1H), 7.54(d, J = 9.3 Hz, 1H), 7.34 (q, 4H), 6.71 (t, J = 4.7 Hz, 1H), 5.42 – 5.30 (m, 8H),4.77 (t, J = 8.2 Hz, 1H), 4.71 (d, J = 9.4 Hz, 1H), 4.57 (dd, J = 15.1, 6.5 Hz,1H), 4.51 (s, 1H), 4.30 (dd, J = 15.1, 5.3 Hz, 1H), 4.01 – 3.88 (m, 3H), 3.73(dd, J = 11.1, 3.6 Hz, 1H), 2.82 – 2.79 (m, 4H), 2.50 (s, 3H), 2.36 – 2.22 (m,3H), 2.22 – 2.18 (m, 2H), 2.06 (dq, J = 14.5, 7.3 Hz, 4H), 1.74 – 1.61 (m, 3H),1.38 – 1.24 (m, 7H), 0.98 (s, 9H), 0.88 (t, J = 7.0 Hz, 3H). 1313C NMR (151 MHz, CDCl3) δ 174.02, 171.38, 171.07, 169.02, 150.38, 148.41, 138.30, 131.57, 130.83, 130.55, 129.44, 128.95, 128.82, 128.67, 128.36, 128.05, 127.86, 127.79, 127.50, 70.18, 58.75, 57.52, 57.50, 43.07, 43.02, 37.49, 35.91, 35.68, 31.51, 29.31, 27.22, 26.69, 26.52, 25.64, 25.62, 25.52, 22.57, 16.06, 14.09. ESI-MS (m / z): [M+Na] calculated for C 44 H 62 N4O5S: 796.4078; found 796.450.

[0101] Example 6

[0102] Referring to the synthesis of Example 5, in Step 3, linoleic acid was used as a raw material for replacement, and the molecule C18U2L1V was synthesized (the structure is shown below).

[0103]

[0104] 1 1H NMR (600 MHz, CDCl3) δ 8.69 (s, 1H), 7.81 (s, 1H), 7.33 (q, J J = 8.2Hz, 4H), 6.65 (s, 1H), 6.36 (s, 1H), 5.41 – 5.27 (m, 4H), 4.75 (t, J J = 8.3 Hz,1H), 4.62 (d, J J = 9.0 Hz, 1H), 4.55 – 4.49 (m, 2H), 4.31 (dd, J J = 15.1, 5.4 Hz,1H), 3.96 (t, J J = 14.6 Hz, 2H), 3.91 – 3.85 (m, 1H), 3.75 – 3.67 (m, 3H), 3.21 – 3.14 (m, 3H), 2.75 (t, J= 6.8 Hz, 2H), 2.50 (s, 3H), 2.27 – 2.22 (m, 1H), 2.21 – 2.14 (m, 2H), 2.03 (p, J = 7.1 Hz, 4H), 1.38 – 1.22 (m, 12H), 1.04 – 0.91 (m, 9H), 0.88 (t, J = 6.8 Hz, 3H). 13 C NMR (150 MHz, CDCl3) δ 174.50, 171.43, 171.30, 169.46, 150.57, 148.30, 138.48, 131.86, 130.82, 130.39, 130.11, 129.56, 128.22, 128.05, 128.00, 70.39, 58.87, 57.81, 57.42, 55.79, 43.73, 43.23, 43.20, 37.36, 36.45, 35.79, 35.77, 31.64, 29.76, 29.46, 29.42, 29.28, 27.33, 26.61, 25.81, 25.76, 22.70, 18.69, 17.29, 17.27, 16.09, 14.21, 12.68. ESI-MS(m / z): [M+Na] calculated for C 42 H 63 N4O5S: 772.455; found: 772.440.

[0105] Example 7

[0106] Referring to the synthesis of Example 5, in Step 3, linolenic acid was used as a raw material for replacement, and a molecule of C18U3L1V was synthesized (the structure is shown below).

[0107]

[0108] 1 H NMR (600 MHz, CDCl3) δ 8.65 (s, 1H), 8.01 (t, J = 6.0 Hz, 1H), 7.49(d, J = 9.2 Hz, 1H), 7.31 (q, J = 8.2 Hz, 4H), 6.78 (t, J= 4.8 Hz, 1H), 5.41 –5.25 (m, 6H), 4.74 (t, J = 8.2 Hz, 1H), 4.67 (d, J = 9.3 Hz, 1H),4.54 – 4.47(m, 2H), 4.29 (dd, J = 15.2, 5.4 Hz, 1H), 3.96 – 3.89 (m, 3H), 3.75 – 3.67 (m,2H), 3.19 – 3.13 (m, 1H), 2.78 (q, J = 5.6 Hz, 3H), 2.48 (s, 3H), 2.25 (d, J =5.4 Hz, 2H), 2.20 – 2.12 (m, 2H), 2.09 – 1.98 (m, 4H), 1.54 (q, J = 7.4 Hz,2H), 1.44 – 1.37 (m, 6H), 1.00 – 0.92 (m, 12H). 13 C NMR (151 MHz, CDCl3) δ174.36, 171.47, 171.01, 169.12, 150.32, 148.41, 138.34, 131.97, 131.56,130.80, 130.17, 129.41, 128.33, 128.20, 127.84, 127.79, 127.09, 70.17, 58.80,57.48, 55.59, 43.53, 43.03, 37.59, 36.31, 35.91, 31.51, 29.61, 29.33, 29.31,29.17, 27.21, 26.52, 25.70, 25.62, 25.53, 20.55, 18.56, 17.15, 16.09, 14.29,12.56. ESI-MS(m / z): [M+Na] calculated for C 42 H 60 N4O5S: 771.3921; found: 771.424.

[0109] Example 8

[0110] Referring to the synthesis of Example 5, in Step 3, cis-5,8,11,14,17-eicosapentaenoic acid was used as a raw material for replacement, and the molecule of C20U5L1V was synthesized (the structure is shown below).

[0111]

[0112] 1 1H NMR (600 MHz, CDCl3) δ 8.67 (s, 1H), 7.96 (t, J J = 6.0 Hz, 1H), 7.46(d, J J = 9.2 Hz, 1H), 7.33 (q, J J = 8.3 Hz, 4H), 6.72 (t, J J = 4.8 Hz, 1H), 5.43 –5.27 (m, 10H), 4.76 (t, J J = 8.2 Hz, 1H), 4.68 (d, J J = 9.2 Hz, 1H), 4.55 (dd, J J =15.1, 6.4 Hz, 1H), 4.51 (s, 1H), 4.31 (dd, J J = 15.1, 5.4 Hz, 1H), 3.99 – 3.88(m, 3H), 3.74 – 3.69 (m, 2H), 3.21 – 3.16 (m, 3H), 2.83 (d, J J = 6.1 Hz, 2H),2.50 (s, 3H), 2.31 – 2.25 (m, 2H), 2.23 – 2.18 (m, 2H), 2.07 (p, J J = 7.4 Hz,4H), 1.65 (p, J J = 7.6 Hz, 2H), 1.42 (d, J J = 6.6 Hz, 4H), 1.01 – 0.94 (m, 12H). 13CNMR (151 MHz, CDCl3) δ 174.05, 171.40, 171.08, 169.08, 150.32, 148.43, 138.30, 132.06, 131.57, 130.82, 129.43, 128.90, 128.86, 128.61, 128.33, 128.27, 128.12, 128.03, 128.03, 127.86, 127.84, 127.00, 70.21, 58.76, 57.56, 57.41, 55.67, 43.60, 43.06, 37.46, 35.83, 35.68, 26.69, 26.51, 25.62, 25.54, 25.53, 20.56, 18.55, 17.14, 16.09, 14.29, 12.58.

[0113] ESI-MS (m / z): [M+K] calculated for C 44 H 60 N4O5S: 794.4182; found: 794.425.

[0114] Example 9

[0115] Referring to the synthesis of Example 5, in Step 3, 13-docosenoic acid was used as a raw material for replacement, and the molecule of C22U1L1V was synthesized (the structure is shown below).

[0116]

[0117] 1 H NMR (600 MHz, CDCl3) δ 8.66 (s, 1H), 7.85 (s, 1H), 7.37 – 7.28 (m, 4H), 6.63 (s, 1H), 5.36 – 5.30 (t, 2H), 4.76 (t, J J = 8.2 Hz, 1H), 4.64 (d, J J = 9.1 Hz, 1H), 4.57 – 4.49 (m, 2H), 4.31 (dd, J J = 15.1, 5.4 Hz, 1H), 4.00 – 3.92 (m, 2H), 3.88 (dd, J= 17.0, 5.0 Hz, 1H), 3.75 – 3.67 (m, 2H), 3.21 – 3.14 (m,1H), 2.50 (s, 3H), 2.37 – 2.29 (m, 1H), 2.28 – 2.21 (m, 1H), 2.18 (t, J = 7.8Hz, 2H), 2.00 (q, J = 6.6 Hz, 4H), 1.36 – 1.21 (m, 28H), 0.97 (s, 9H), 0.87 (t, J = 7.0 Hz, 3H). 13 C NMR (151 MHz, CDCl3) δ 174.50, 171.37, 171.30, 169.38,150.42, 148.60, 138.38, 131.68, 131.02, 130.07, 129.99, 129.58, 128.04,70.39, 58.82, 57.78, 57.48, 55.84, 43.79, 43.23, 37.38, 36.50, 35.81, 32.04,29.94, 29.91, 29.79, 29.74, 29.66, 29.52, 29.49, 29.46, 27.36, 26.63, 25.83,22.82, 18.74, 17.37, 16.23, 14.26, 12.63. ESI-MS(m / z): [M+Na] calculated forC 46 H 73 N5O5S (M+Na): 830.533, found: 830.521.

[0118] Example 10

[0119] Referring to the synthesis of Example 5, in Step 2, N-Boc-γ-aminobutyric acid was used as a raw material for replacement, and the molecule of C20U4L3V was synthesized (the structure is shown below).

[0120]

[0121] 1 H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.44 (t, J = 6.0 Hz, 1H), 7.35(q, 4H), 7.00 (d, J= 8.3 Hz, 1H), 6.01 (t, J = 5.9 Hz, 1H), 5.46 – 5.28 (m, 8H),4.74 (t, J = 8.1 Hz, 1H), 4.57 – 4.48 (m, 3H), 4.33 (dd, J = 15.0, 5.4 Hz, 1H),4.09 (d, J = 11.3 Hz, 1H), 3.61 (dd, J = 11.3, 3.6 Hz, 1H), 3.33 – 3.17 (m, 2H),2.83 – 2.78 (m, 4H), 2.51 (s, 3H), 2.27 – 2.11 (m, 7H), 2.12 – 2.02 (m, 4H),1.79 – 1.72 (m, 2H), 1.68 (p, J = 7.5 Hz, 2H), 1.39 – 1.24 (m, 7H), 0.96 (s,9H), 0.88 (t, J = 6.9 Hz, 3H). 13 C NMR (151 MHz, CDCl3) δ 173.71, 173.29, 172.00,170.90, 150.32, 148.47, 138.16, 131.59, 130.95, 130.55, 129.51, 129.06,128.81, 128.65, 128.31, 128.11, 128.08, 127.82, 127.50, 70.13, 58.44, 58.12,56.85, 43.19, 38.46, 36.19, 36.07, 34.95, 33.18, 31.51, 29.32, 27.23, 26.70,26.48, 26.03, 25.65, 25.63, 25.58, 22.57, 16.06, 14.09. ESI-MS(m / z): [M+K]calculated for C 46 H 66 N4O5S (M+K): 824.4652, found: 824.483.

[0122] Example 11

[0123] Referring to the synthesis in Example 5, in Step 2, N-Boc-γ-aminobutyric acid was used as a raw material for replacement, and in Step 3, oleic acid was used as a raw material for replacement, and a molecule of C18U1L3V was synthesized (the structure is shown below).

[0124]

[0125] 1 H NMR (600 MHz, DMSO-d6) δ 8.98 (s, 1H), 8.56 (t, J = 6.1 Hz, 1H),7.90 (d, J = 9.3 Hz, 1H), 7.73 (t, J = 5.6 Hz, 1H), 7.45 – 7.36 (m, 4H), 5.31(t, J = 4.9 Hz, 2H), 5.13 (d, J = 3.6 Hz, 1H), 4.53 (d, J = 9.3 Hz, 1H), 4.46– 4.40 (m, 2H), 4.35 (s, 1H), 4.21 (dd, J = 15.8, 5.5 Hz, 1H), 3.67 (dd, J =10.5, 4.1 Hz, 1H), 3.63 (d, J = 10.8 Hz, 1H), 3.16 – 3.12 (m, 1H), 3.05 –2.95 (m, 2H), 2.44 (s, 3H), 2.28 – 2.20 (m, 1H), 2.16 – 2.08 (m, 1H), 2.02(t, J = 7.4 Hz, 2H), 1.97 (q, J = 6.5 Hz, 4H), 1.93 – 1.87 (m, 1H), 1.64 –1.53 (m, 2H), 1.46 (p, J = 7.4 Hz, 2H), 1.31 – 1.18 (m, 20H), 0.93 (s, 3H),0.84 (t, J = 6.9 Hz, 3H). 1313C NMR (150 MHz, DMSO-d6) δ 172.46, 172.42, 172.23, 170.13, 139.97, 131.63, 130.10, 129.11, 127.89, 69.35, 59.17, 56.84, 56.83, 54.07, 42.32, 42.12, 38.57, 38.41, 35.91, 35.72, 33.01, 31.74, 29.59, 29.55, 29.28, 29.16, 29.14, 29.04, 29.01, 27.06, 27.03, 26.85, 26.26, 25.76, 22.55, 18.56, 17.20, 16.41, 14.42, 12.96. ESI-MS(m / z): [M+Na] calculated for C 44 H 69 N5O5S (M+K): 802.4917, found: 802.490.

[0126] Example 12

[0127] Referring to the synthesis of Example 5, in Step 2, N-Boc-γ-aminobutyric acid was used as a raw material for replacement, and in Step 3, linoleic acid was used as a raw material for replacement to synthesize the molecule of C18U2L3V (the structure is shown below).

[0128]

[0129] 1 1H NMR (600 MHz, DMSO) δ 8.98 (s, 1H), 8.56 (t, J J = 6.1 Hz, 1H), 7.89(d, J J = 9.3 Hz, 1H), 7.73 (t, J J = 5.6 Hz, 1H), 7.46 – 7.37 (m, 4H), 5.38 – 5.25(m, 4H), 5.13 (d, J J = 3.6 Hz, 1H), 4.53 (d, J J = 9.4 Hz, 2H), 4.46 – 4.40 (m, 2H),4.35 (s, 1H), 4.21 (dd, J= 15.8, 5.6 Hz, 1H), 3.70 – 3.60 (m, 2H), 3.05 – 2.94(m, 2H), 2.73 (t, J = 6.8 Hz, 2H), 2.44 (s, 3H), 2.28 – 2.20 (m, 1H), 2.16 –2.07 (m, 1H), 2.01 (p, J = 7.3 Hz, 6H), 1.94 – 1.86 (m, 1H), 1.66 – 1.52 (m,1H), 1.33 – 1.18 (m, 16H), 0.93 (s,9H), 0.85 (t, J = 6.9 Hz, 3H). 13 C NMR (150 MHz, DMSO) δ 172.47, 172.42, 172.24, 170.13, 151.92, 148.18, 139.97, 131.64, 130.21, 130.19, 130.10, 129.11, 128.22, 127.89, 70.25, 69.34, 59.17, 56.84, 42.12, 40.51, 38.58, 38.41, 35.91, 35.72, 33.01, 31.36, 29.50, 29.18, 29.14, 29.14, 29.04, 27.10, 27.07, 26.85, 26.27, 25.77, 25.69, 22.44, 16.41, 14.40. ESI-MS (m / z): [M+Na] calculated for C 44 H 67 N5O5S (M+Na): 800.486, found: 800.474.

[0130] Example 13

[0131] Referring to the synthesis of Example 5, in Step 2, N-Boc-γ-aminobutyric acid was used as a raw material for replacement, and in Step 3, linolenic acid was used as a raw material for replacement to synthesize the molecule of C18U3L3V (the structure is shown below).

[0132]

[0133] 1 H NMR (600 MHz, CDCl3) δ 8.66 (s, 1H), 7.47 (t, J= 6.0 Hz, 1H), 7.32(q, J = 8.3 Hz, 4H), 7.06 (d, J = 8.4 Hz, 1H), 6.16 (t, J = 6.0 Hz, 1H), 5.42 –5.25 (m, 6H), 4.71 (t, J = 8.1 Hz, 1H), 4.56 – 4.45 (m, 3H), 4.32 (dd, J = 15.1,5.5 Hz, 1H), 3.61 (dd, J = 11.3, 3.6 Hz, 1H), 3.29 – 3.12 (m, 2H), 2.81 – 2.75(m, 3H), 2.49 (s, 3H), 2.44 – 2.37 (m, 1H), 2.25 – 2.10 (m, 5H), 2.10 – 1.99(m, 4H), 1.72 (p, J = 7.0 Hz, 2H), 1.56 (p, J = 8.6 Hz, 2H), 1.34 – 1.22 (m,10H), 0.95 (d, J = 3.6 Hz, 12H). 13 C NMR (151 MHz, CDCl3) δ 174.05, 173.35,171.87, 171.06, 150.34, 148.42, 138.20, 131.97, 131.60, 130.88, 130.21,129.47, 128.31, 128.21, 128.01, 127.76, 127.09, 70.11, 58.62, 58.09, 56.89,43.14, 38.52, 36.71, 36.45, 35.08, 33.17, 29.61, 29.32, 29.30, 29.17, 27.20,26.48, 25.93, 25.80, 25.61, 25.52, 20.55, 16.05, 14.29. ESI-MS(m / z): [M+Na]calculated for C 44 H 64 N5O4S 760.4597, found:798.4234.

[0134] Example 14

[0135] Referring to the synthesis in Example 5, in Step 2, N-Boc-γ-aminobutyric acid was used as a raw material for replacement, and in Step 3, cis-5,8,11,14,17-eicosapentaenoic acid was used as a raw material for replacement, and the molecule of C20U5L3V was synthesized (the structure is shown below).

[0136]

[0137] 1 H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.46 (t, J J = 6.1 Hz, 1H), 7.34(q, J J = 8.4 Hz, 4H), 7.08 (d, J J = 8.4 Hz, 1H), 6.16 (t, J J = 5.9 Hz, 1H), 5.42 –5.31 (m, 10H), 4.73 (t, J J = 8.1 Hz, 1H), 4.57 – 4.47 (m, 3H), 4.34 (dd, J J = 15.1,5.5 Hz, 1H), 4.07 (d, J J = 11.5 Hz, 1H), 3.71 (p, J J = 6.7 Hz, 1H), 3.63 (dd, J J=11.3, 3.6 Hz, 1H), 3.30 – 3.15 (m, 3H), 2.51 (s, 3H), 2.47 – 2.42 (m, 1H),2.36 – 2.30 (m, 1H), 2.26 – 2.19 (m, 2H), 2.19 – 2.14 (m, 4H), 2.07 (pt, J J=7.6, 3.1 Hz, 4H), 1.78 – 1.71 (m, 2H), 1.71 – 1.64 (m, 2H), 1.47 – 1.41 (m,4H), 0.99 – 0.94 (m, 12H). 1313C NMR (151 MHz, CDCl3) δ 173.78, 173.37, 171.91, 171.05, 150.37, 148.40, 138.20, 132.07, 131.63, 130.87, 129.48, 129.08, 128.75, 128.61, 128.31, 128.22, 128.17, 128.05, 128.03, 127.84, 126.99, 70.12, 58.60, 58.10, 56.89, 55.48, 43.45, 43.15, 38.55, 36.41, 36.05, 35.05, 33.17, 26.70, 26.47, 25.91, 25.63, 25.60, 25.54, 20.56, 16.03, 14.29.

[0138] ESI-MS (m / z): [M+Na] calculated for C 46 H 64 N4O5S 822.4234, found: 822.458.

[0139] Example 15

[0140] Referring to the synthesis of Example 5, in Step 2, N-Boc-γ-aminobutyric acid was used as a raw material for replacement, and in Step 3, 13-docosenoic acid was used as a raw material for replacement to synthesize the molecule of C22U1L3V (the structure is shown below).

[0141]

[0142] 1 1H NMR (600 MHz, DMSO) δ 8.98 (s, 1H), 8.56 (t, J J = 6.1 Hz, 1H), 7.90 (d, J J = 9.3 Hz, 1H), 7.73 (t, J J = 5.6 Hz, 1H), 7.39 (q, 4H), 5.36 – 5.29 (m, 2H), 5.13 (d, J J = 3.5 Hz, 1H), 4.54 (d, J J = 9.3 Hz, 1H), 4.43 (d, 2H), 4.35 (s, 1H), 4.22 (dd, J= 15.8, 5.5 Hz, 1H), 3.69 – 3.61 (m, 2H), 3.05 – 2.96 (m, 2H), 2.44(s, 3H), 2.29 – 2.20 (m, 1H), 2.15 – 2.09 (m, 1H), 2.03 (t, J = 7.4 Hz, 3H),1.97 (q, J = 6.5 Hz, 3H), 1.93 – 1.87 (m, 1H), 1.64 – 1.54 (m, 2H), 1.50 – 1.42(m, 2H), 1.32 – 1.18 (m, 28H), 0.94 (s, 9H), 0.85 (t, J = 6.9 Hz, 3H). 13 C NMR(151 MHz, DMSO) δ 172.47, 172.42, 172.24, 170.13, 151.92, 148.18, 139.97,131.63, 130.11, 129.10, 127.89, 69.35, 59.17, 56.85, 42.12, 40.51, 38.57,38.41, 35.92, 35.72, 33.01, 31.75, 29.55, 29.49, 29.43, 29.33, 29.29, 29.16,29.04, 27.02, 26.85, 26.26, 25.77, 22.56, 16.41, 14.41. ESI-MS(m / z):[M+Na]calculated for C 48 H 77 N5O5S 858.565, found:858.553.

[0143] Example 16

[0144] Referring to the synthesis of Example 5, in Step 2, N-Boc-5-aminovaleric acid was used as a raw material for replacement, and a molecule of C20U4L4V was synthesized (the structure is shown below).

[0145]

[0146] 1 H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.41 – 7.32 (m, 5H), 6.39 (d, J= 8.7 Hz, 1H), 5.80 (t, J = 5.9 Hz, 1H), 5.44 – 5.30 (m, 8H), 4.72 (t, J = 8.1Hz, 1H), 4.59 – 4.50 (m, 3H), 4.34 (dd, J = 15.0, 5.4 Hz, 1H), 4.07 (d, J = 11.3Hz, 1H), 3.62 (dd, J = 11.3, 3.5 Hz, 1H), 3.19 (q, J = 6.6 Hz, 2H), 2.83 – 2.79(m, 4H), 2.51 (s, 3H), 2.29 – 2.23 (m, 1H), 2.21 – 2.12 (m, 5H), 2.11 – 2.03(m, 5H), 1.72 – 1.57 (m, 5H), 1.51 – 1.44 (m, 2H), 1.39 – 1.33 (m, 2H), 1.32– 1.24 (m, 5H), 0.94 (s, 9H), 0.88 (t, J = 6.8 Hz, 3H). 13 C NMR (151 MHz, CDCl3)δ 173.48, 173.29, 171.89, 170.87, 150.32, 148.48, 138.15, 131.59, 130.96,130.55, 129.52, 129.12, 128.76, 128.64, 128.28, 128.15, 128.08, 127.84,127.51, 70.08, 58.56, 57.62, 56.87, 43.22, 38.74, 36.18, 36.10, 35.52, 35.08,31.52, 29.32, 28.84, 27.23, 26.70, 26.43, 25.65, 25.64, 25.57, 22.58, 22.51,16.07, 14.09. ESI-MS(m / z): [M+Na] calculated for C 47 H 68 N4O5S 838.4910, found:838.501.

[0147] Example 17

[0148] Referring to the synthesis in Example 5, in Step 2, N-Boc-5-aminovaleric acid was used as a raw material for replacement, and in Step 3, linoleic acid was used as a raw material for replacement, and the molecule of C18U2L4V was synthesized (the structure is shown below).

[0149]

[0150] 1 H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.41 (t, J J = 6.0 Hz, 1H), 7.35(q, J J = 8.3 Hz, 4H), 6.43 (d, J J = 8.7 Hz, 1H), 5.89 (t, J J = 5.9 Hz, 1H), 5.42 –5.28 (m, 4H), 4.72 (t, J J = 8.1 Hz, 1H), 4.58 – 4.49 (m, 3H), 4.35 (dd, J J = 15.0,5.4 Hz, 1H), 4.06 (d, J J = 11.5 Hz, 1H), 4.00 (s, 1H), 3.70 (p, J J = 6.7 Hz, 1H),3.62 (dd, J J = 11.3, 3.6 Hz, 1H), 3.22 – 3.11 (m, 3H), 2.76 (t, J J = 6.8 Hz, 2H),2.51 (s, 3H), 2.49 – 2.44 (m, 1H), 2.28 – 2.23 (m, 1H), 2.22 – 2.11 (m,6H),2.08 – 2.00 (m, 4H), 1.58 (t, J J = 6.8 Hz, 2H), 1.38 – 1.24 (m, 16H), 0.94 (s,9H), 0.89 (t, J J = 6.9 Hz, 3H). 1313C NMR (151 MHz, CDCl3) δ 173.78, 173.65, 171.95, 171.05, 150.42, 148.57, 138.28, 131.71, 131.04, 130.36, 130.14, 129.61, 128.17, 128.00, 70.19, 58.71, 57.75, 56.98, 55.12, 43.31, 43.17, 38.83, 36.88, 36.36, 35.62, 35.20, 31.63, 29.75, 29.45, 29.42, 29.29, 28.92, 27.31, 26.54, 25.91, 25.74, 22.68, 22.65, 16.18, 14.20, 12.49. ESI-MS (m / z): [M+Na] calculated for C 45 H 69 N5O5S 814.502, found: 814.490.

[0151] Example 18

[0152] Referring to the synthesis of Example 5, in Step 2, N-Boc-5-aminovaleric acid was used as a raw material for replacement, and in Step 3, linolenic acid was used as a raw material for replacement to synthesize the molecule of C18U3L4V (the structure is shown below).

[0153]

[0154] 1 1H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.45 – 7.41 (m, 1H), 7.35 (q, 4H), 6.44 (t, J J = 9.8 Hz, 1H), 5.88 (t, J J = 6.5 Hz, 1H), 5.42 – 5.28 (m, 6H), 4.72 (t, J J = 7.9 Hz, 1H), 4.58 – 4.50 (m, 3H), 4.34 (dd, J J = 15.0, 5.4 Hz, 1H), 4.05 (d, J J = 11.4 Hz, 2H), 3.63 (dd, J J = 11.2, 3.6 Hz, 1H), 3.19 (q, J= 6.6 Hz, 2H), 2.83 – 2.77 (m, 4H), 2.51 (s, 3H), 2.49 – 2.44 (m, 1H), 2.27 – 2.23 (m, 1H), 2.21 – 2.11 (m, 5H), 2.05 (dt, J = 23.7, 7.1 Hz, 4H), 1.63 – 1.55 (m, 3H), 1.52 – 1.44 (m, 2H), 1.36 – 1.31 (m, 2H), 1.30 – 1.27 (m, 6H), 0.97 (t, J = 7.5 Hz, 3H), 0.94 (s, 9H). 13 C NMR (151 MHz, CDCl3) δ 173.59, 173.45, 171.85, 170.95, 150.31, 148.46, 138.18, 131.97, 131.60, 130.93, 130.24, 129.50, 128.30, 128.23, 128.05, 127.74, 127.10, 70.05, 58.61, 57.61, 56.86, 43.19, 40.98, 38.71, 36.77, 36.26, 35.51, 35.15, 29.61, 29.30, 29.16, 28.84, 27.21, 26.43, 25.79, 25.62, 25.53, 22.54, 20.55, 16.06, 14.29. ESI-MS (m / z): [M+Na] calculated for C 45 H 67 N5O5S 812.4761, found: 812.473.

[0155] Example 19

[0156] Referring to the synthesis of Example 5, in Step 2, N-Boc-5-aminovaleric acid was used as a raw material for replacement, and in Step 3, cis-5,8,11,14,17-eicosapentaenoic acid was used as a raw material for replacement to synthesize the molecule of C20U5L4V (the structure is shown below).

[0157]

[0158] 1 H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.42 (t,J = 6.0 Hz, 1H), 7.35(q, J = 8.4 Hz, 4H), 6.44 (d, J = 8.8 Hz, 1H), 5.85 (t, J = 5.9 Hz, 1H), 5.55 –5.17 (m, 10H), 4.71 (t, J = 8.1 Hz, 1H), 4.59 – 4.49 (m, 4H), 4.34 (dd, J = 15.0,5.3 Hz, 1H), 4.06 (d, J = 11.8 Hz, 1H), 3.63 (dd, J = 11.2, 3.6 Hz, 1H), 3.19 (q, J = 6.6 Hz, 2H), 2.84 – 2.76 (m, 4H), 2.51 (s, 3H), 2.49 – 2.44 (m, 1H), 2.25(p, J = 7.2 Hz, 2H), 2.21 – 2.12 (m, 5H), 2.11 – 2.03 (m, 4H), 1.72 – 1.55 (m,5H), 1.47 (dq, J = 14.0, 6.7 Hz, 2H), 0.99 – 0.93 (m, 12H). 13 C NMR (151 MHz,CDCl3) δ 173.48, 173.31, 171.84, 170.94, 150.34, 148.45, 138.17, 132.07,131.64, 131.60, 130.93, 129.50, 129.14, 128.72, 128.61, 128.31, 128.20,128.06, 127.85, 127.00, 70.07, 58.61, 57.61, 56.88, 43.20, 40.96, 38.75,36.26, 36.09, 35.51, 35.13, 28.85, 26.70, 26.43, 25.64, 25.57, 25.55, 22.54,20.57, 16.06, 14.29. ESI-MS(m / z):[M+Na] calculated for C 45 H 67N5O5S 852.4500., found: 852.508.

[0159] Example 20

[0160] Referring to the synthesis of Example 5, in Step 2, N-Boc-5-aminovaleric acid was used as a raw material for replacement, and in Step 3, 13-docosenoic acid was used as a raw material for replacement to synthesize the molecule of C22U1L4V (the structure is shown below).

[0161]

[0162] 1 H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.39 (t, J J = 6.0 Hz, 1H), 7.35 (q, J J = 8.3 Hz, 4H), 6.40 (d, J J = 8.8 Hz, 1H), 5.81 (t, J J = 5.9 Hz, 1H), 5.34 (t, 2H), 4.71 (t, J J = 8.1 Hz, 1H), 4.58 – 4.49 (m, 3H), 4.34 (dd, J J = 15.0, 5.3 Hz, 1H), 4.07 (d, J J = 11.5 Hz, 1H), 3.61 (dd, J J = 11.3, 3.6 Hz, 1H), 3.19 (q, J J = 6.6 Hz, 2H), 2.51 (s, 3H), 2.27 – 2.23 (m, 1H), 2.21 – 2.10 (m, 4H), 2.00 (q, J = 6.6 Hz, 4H), 1.64 (dt, J J = 14.4, 7.3 Hz, 1H), 1.62 – 1.53 (m, 3H), 1.51 – 1.39 (m, 3H), 1.33 – 1.22 (m, 28H), 0.93 (s, 9H), 0.87 (t, J J = 6.9 Hz, 3H). 1313C NMR (151 MHz, CDCl3) δ 173.68, 173.56, 171.90, 170.88, 150.36, 148.38, 138.19, 131.66, 130.90, 129.93, 129.88, 129.51, 128.09, 70.09, 58.56, 57.65, 56.88, 43.22, 38.70, 36.81, 36.17, 35.51, 35.05, 32.62, 31.91, 29.80, 29.78, 29.72, 29.69, 29.66, 29.59, 29.55, 29.53, 29.50, 29.41, 29.37, 29.35, 29.33, 28.82, 27.23, 26.43, 25.81, 22.69, 22.51, 16.03, 14.13. ESI-MS (m / z): [M+Na] calculated for C 49 H 79 N5O5S 872.580, found: 872.567.

[0163] Example 21

[0164] Referring to the synthesis of Example 5, in Step 2, (2-aminoethoxy)acetic acid was used as a raw material for replacement, and a molecule of C20U4L4OV was synthesized (the structure is shown below).

[0165]

[0166] 1 1H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.35 (q, 4H), 7.13 (d, J J = 8.8 Hz, 1H), 6.18 (t, 1H), 5.44 – 5.29 (m, 8H), 4.72 (t, J J = 7.9 Hz, 1H), 4.57 – 4.50 (m, 3H), 4.37 (dd, J J = 15.0, 5.4 Hz, 1H), 4.08 – 3.98 (m, 2H), 3.89 (d, J= 15.5 Hz, 1H), 3.69 – 3.61 (m, 2H), 3.56 – 3.51 (m, 1H), 3.50 – 3.39 (m, 2H), 3.34 (s, 1H), 2.83 – 2.79 (m, 4H), 2.52 (s, 3H), 2.20 (t, 2H), 2.16 – 2.11 (m, 1H), 2.11 – 2.02 (m, 4H), 1.86 (s, 2H), 1.70 (p, J = 7.0 Hz, 2H), 1.36 (q, J = 7.3 Hz, 2H), 1.32 – 1.25 (m, 5H), 0.95 (s, 9H), 0.88 (t, J = 6.9 Hz, 3H). 13 13C NMR (151 MHz, CDCl3) δ 173.32, 171.55, 170.58, 170.03, 150.35, 148.53, 138.01, 131.51, 131.11, 130.56, 129.56, 129.11, 128.80, 128.66, 128.32, 128.14, 127.83, 127.51, 70.82, 70.18, 70.13, 58.53, 57.04, 56.83, 43.28, 39.16, 35.93, 35.87, 35.14, 31.52, 29.33, 27.23, 26.70, 26.41, 25.65, 25.64, 25.47, 22.58, 16.08, 14.10. ESI-MS (m / z): [M+Na] calculated for C 46 H 67 N5O6S840.4710, found: 840.471.

[0167] Example 22

[0168] Referring to the synthesis of Example 5, in Step 2, (2-aminoethoxy)acetic acid was used as a raw material for replacement, and in Step 3, linoleic acid was used as a raw material for replacement to synthesize the molecule of C18U2L4OV (the structure is shown below).

[0169]

[0170] 11H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.45 (q, J = 5.8 Hz, 1H),7.38 – 7.28 (m, 4H), 7.18 (d, J = 9.1 Hz, 1H), 6.52 (dd, J = 20.5, 8.9 Hz,1H), 5.42 – 5.27 (m, 1H), 4.70 (t, J = 7.9 Hz, 1H), 4.56 (d, J = 9.1 Hz, 1H),4.54 – 4.46 (m, 2H), 4.39 (dd, J = 14.9, 5.6 Hz, 1H), 4.00 – 3.93 (m, 2H),3.88 (d, J = 15.5 Hz, 1H), 3.68 (dd, J = 11.1, 3.9 Hz, 1H), 3.66 – 3.61 (m,1H), 3.55 – 3.44 (m, 2H), 3.41 – 3.35 (m, 1H), 2.75 (q, J = 5.5 Hz, 2H), 2.51(s, 3H), 2.45 – 2.38 (m, 1H), 2.22 – 2.12 (m, 3H), 2.04 (q, J = 7.3 Hz, 4H),1.42 (m, 3H), 1.36 – 1.24 (m, 14H), 0.96 (s, 9H), 0.88 (t, J = 7.0 Hz, 3H). 13 13C NMR (151 MHz, CDCl3) δ 173.86, 173.84, 171.18, 171.04, 169.84, 150.37,148.41, 138.17, 138.15, 131.56, 130.92, 130.23, 129.98, 129.46, 128.06,128.00, 127.87, 70.69, 70.03, 69.98, 58.87, 56.90, 56.85, 55.37, 43.13,39.05, 36.51, 35.58, 31.49, 29.64, 29.35, 29.32, 27.19, 26.39, 25.73, 25.62,22.56, 18.56, 17.18, 16.05, 14.08, 12.54. ESI-MS(m / z): [M+Na] calculated forC 44 H 67N5O6S 816.4710, found: 816.471.

[0171] Example 23

[0172] Referring to the synthesis of Example 5, in Step 2, (2-aminoethoxy)acetic acid was used as a raw material for replacement, and in Step 3, linolenic acid was used as a raw material for replacement to synthesize a molecule of C18U3L4OV (the structure is shown below).

[0173]

[0174] 1 H NMR (600 MHz, CDCl3) δ 8.65 (s, 1H), 7.39 (d, J = 6.0 Hz, 1H), 7.32 (q, J = 7.8 Hz, 4H), 7.14 (d, J = 9.0 Hz, 1H), 6.47 – 6.39 (m, 1H), 5.40 – 5.25 (m, 6H), 4.67 (m, 1H), 4.53 (d, J = 9.1 Hz, 1H), 4.51 – 4.46 (m, 2H), 4.36 (dd, J = 15.0, 5.6 Hz, 1H), 3.97 – 3.92 (m, 2H), 3.86 (d, J = 15.5 Hz, 1H), 3.66 – 3.59 (m, 2H), 3.53 – 3.41 (m, 2H), 3.38 – 3.33 (m, 1H), 3.17 – 3.13 (m, 1H), 2.77 (q, J = 5.6 Hz, 3H), 2.48 (s, 3H), 2.43 – 2.37 (m, 1H), 2.15 (t, J = 7.6 Hz, 2H), 2.06 – 1.98 (m, 4H), 1.41 – 1.36 (m, 4H), 1.32 – 1.23 (m, 8H), 0.96 – 0.90 (m, 12H). 1313C NMR (151 MHz, CDCl3) δ 173.84, 171.22, 170.99, 169.88, 150.36, 148.43, 138.14, 131.98, 131.56, 130.92, 130.21, 129.47, 128.30, 128.19, 128.02, 127.72, 127.09, 70.69, 70.07, 70.00, 58.84, 56.93, 56.84, 55.46, 43.15, 39.07, 36.52, 35.53, 31.50, 29.62, 29.35, 29.33, 29.17, 27.21, 26.40, 25.73, 25.61, 25.52, 20.54, 18.56, 17.16, 16.05, 14.29, 12.56. ESI-MS (m / z): [M+Na] calculated for C 44 H 65 N5O6S 814.4553, found: 814.450.

[0175] Example 24

[0176] Referring to the synthesis of Example 5, in Step 2, (2-aminoethoxy)acetic acid was used as a raw material for replacement, and in Step 3, cis-5,8,11,14,17-eicosapentaenoic acid was used as a raw material for replacement, and the molecule C20U5L4OV was synthesized (the structure is shown below).

[0177]

[0178] 1 1H NMR (600 MHz, CDCl3) δ 8.65 (s, 1H), 7.38 (t, J J = 6.0 Hz, 1H), 7.32 (q, J J = 8.3 Hz, 4H), 7.14 (d, J J = 9.1 Hz, 1H), 6.42 (t, J J = 5.8 Hz, 1H), 5.43 – 5.22 (m, 10H), 4.67 (t, J J = 7.9 Hz, 1H), 4.55 – 4.45 (m, 3H), 4.36 (dd, J= 15.1, 5.7 Hz, 1H), 4.09 (s, 1H), 3.97 – 3.90 (m, 2H), 3.86 (d, J = 15.5 Hz, 1H), 3.71– 3.57 (m, 3H), 3.52 – 3.42 (m, 2H), 3.40 – 3.32 (m, 1H), 3.15 (q, J = 7.5 Hz, 1H), 2.76 – 2.74 (m, 2H), 2.48 (s, 3H), 2.42 – 2.36 (m, 1H), 2.17 (t, 2H), 2.15 – 2.10 (m, 1H), 2.04 (m, 4H), 1.68 – 1.62 (m, 2H), 1.43 – 1.37 (m, 4H), 0.96 – 0.92 (m, 12H). 13 C NMR (151 MHz, CDCl3) δ 173.52, 171.19, 171.02, 169.86, 150.38, 148.42, 138.14, 132.04, 131.56, 130.92, 129.47, 129.10, 128.72, 128.59, 128.31, 128.21, 128.18, 128.04, 128.02, 127.83, 126.99, 70.65, 70.05, 70.00, 58.85, 56.94, 56.83, 55.42, 43.38, 43.14, 39.08, 36.48, 35.84, 35.53, 26.69, 26.40, 25.62, 25.53, 25.52, 20.53, 18.56, 17.16, 16.02, 14.30, 12.55. ESI-MS (m / z): [M+Na] calculated for C 46 H 65 N5O6S 838.4553, found: 838.453.

[0179] Example 25

[0180] Referring to the synthesis of Example 5, in Step 2, N-Boc-6-aminohexanoic acid was used as a raw material for replacement, and the molecule C20U4L5V was synthesized (the structure is shown below).

[0181]

[0182] 1 1H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.44 (t, J J = 6.0 Hz, 1H), 7.38 –7.33 (m, 4H), 6.30 (d, J J = 8.9 Hz, 1H), 5.74 (t, J J = 5.8 Hz, 1H), 5.43 – 5.30 (m,8H), 4.72 (t, J J = 8.1 Hz, 1H), 4.58 – 4.49 (m, 3H), 4.35 (dd, J J = 15.0, 5.4 Hz,1H), 4.05 (d, J J = 11.5 Hz, 1H), 3.94 (s, 1H), 3.62 (dd, J J = 11.2, 3.6 Hz, 1H),3.19 (q, J J = 6.4 Hz, 2H), 2.83 – 2.79 (m, 4H), 2.51 (s, 3H), 2.49 – 2.44 (m,1H), 2.24 – 2.11 (m, 6H), 2.12 – 2.02 (m, 4H), 1.68 (p, J J = 7.5 Hz, 2H), 1.64 –1.55 (m, 2H), 1.46 (p, J J = 7.4 Hz, 2H), 1.40 – 1.23 (m, 9H), 0.94 (s, 9H), 0.88(t, J J = 7.0 Hz, 3H). 1313C NMR (151 MHz, CDCl3) δ 173.51, 173.02, 171.82, 170.95, 150.32, 148.46, 138.17, 131.60, 130.93, 130.54, 129.50, 129.13, 128.74, 128.63, 128.27, 128.14, 128.06, 127.83, 127.51, 70.00, 58.63, 57.48, 56.84, 43.19, 39.25, 36.23, 36.11, 35.11, 31.51, 29.32, 29.30, 27.22, 26.69, 26.42, 26.21, 25.65, 25.63, 25.58, 25.09, 22.57, 16.06, 14.09. ESI-MS (m / z): [M+K] calculated for C 48 H 70 N4O5S 852.4704, found: 852.508.

[0183] Example 26

[0184] Referring to the synthesis of Example 5, in Step 2, N-Boc-6-aminohexanoic acid was used as a raw material for replacement, and in Step 3, linoleic acid was used as a raw material for replacement to synthesize the molecule of C18U2L5V (the structure is shown below).

[0185]

[0186] 1 1H NMR (600 MHz, CDCl3) δ 8.64 (s, 1H), 7.48 (t, J J = 7.2 Hz, 1H), 7.31(q, J = 2.8 Hz, 4H), 6.45 – 6.39 (m, 1H), 6.00 (p, J J = 5.6 Hz, 1H), 5.38 – 5.26(m, 4H), 4.67 (t, J J = 7.2 Hz, 1H), 4.51 – 4.45 (m, 2H), 4.33 (dd, J J = 15.1, 5.5Hz, 1H), 4.28 – 4.22 (m, 1H), 3.97 (d, J J = 11.5 Hz, 1H), 3.62 (dd, J= 11.1, 3.8 Hz, 1H), 3.15 (q, J = 6.6 Hz, 2H), 2.73 (t, J = 6.9 Hz, 2H), 2.47 (s, 3H), 2.40 – 2.32 (m, 1H), 2.17 – 2.06 (m, 4H), 2.00 (p, J = 6.8 Hz, 4H), 1.59 – 1.50 (m, 4H), 1.40 – 1.36 (m, 4H), 1.30 – 1.22 (m, 14H), 0.92 (s, 9H), 0.85 (t, J = 7.0 Hz, 3H). 13 C NMR (151 MHz, CDCl3) δ 173.49, 173.46, 171.65, 171.19, 150.34, 148.37, 138.25, 131.64, 130.81, 130.22, 130.00, 129.43, 128.04, 127.97, 127.87, 69.96, 58.82, 57.51, 56.82, 55.36, 53.47, 43.35, 43.10, 39.22, 36.74, 36.55, 36.06, 35.28, 31.50, 29.63, 29.32, 29.30, 29.19, 29.17, 27.19, 26.43, 26.24, 25.85, 25.62, 25.12, 22.56, 18.55, 17.12, 16.05, 14.08, 12.62. ESI-MS (m / z): [M+K] calculated for C 46 H 70 N4O5S 828.4704, found: 828.506.

[0187] Example 27

[0188] Referring to the synthesis of Example 5, in Step 2, N-Boc-6-aminohexanoic acid was used as a raw material for replacement, and in Step 3, linolenic acid was used as a raw material for replacement to synthesize the molecule of C18U2L5V (the structure is shown below).

[0189]

[0190] 11H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.46 (t, J J = 6.0 Hz, 1H), 7.37 –7.33 (m, 4H), 6.43 (d, J = 8.7 Hz, 2H),6.16 (q, J J = 5.2 Hz, 1H), 5.43 – 5.27(m, 6H), 4.69 (t, J J = 8.1 Hz, 1H), 4.54 – 4.48 (m, 3H), 4.37 (dd, J J = 15.1, 5.6Hz, 1H), 4.00 (d, J J = 11.2 Hz, 1H), 3.75 – 3.63 (m, 4H), 3.23 – 3.12 (m, 6H),2.80 (q, J J = 5.3 Hz, 4H), 2.50 (s, 3H), 2.41 – 2.35 (m, 1H), 2.25 – 2.11 (m,5H), 2.11 – 2.00 (m, 4H), 1.62 – 1.53 (m, 4H), 1.47 (q, J J = 7.4 Hz, 2H), 1.34 –1.25 (m, 12H), 1.02 – 0.92 (m, 12H). 1313C NMR (151 MHz, CDCl3) δ 173.74, 173.62, 171.64, 171.29, 150.37, 148.33, 138.23, 131.96, 131.68, 130.76, 130.23, 129.41, 128.29, 128.22, 127.94, 127.71, 127.08, 70.01, 58.88, 57.64, 56.76, 55.58, 43.52, 43.07, 39.28, 36.72, 36.63, 36.04, 35.19, 31.49, 29.62, 29.59, 29.30, 29.25, 29.16, 29.02, 27.20, 26.42, 26.19, 25.92, 25.62, 25.60, 25.51, 25.15, 22.55, 20.54, 18.47, 16.98, 16.05, 14.28, 12.71. ESI-MS (m / z): [M+K] calculated for C 46 H 68 N4O5S 826.4547, found: 826.489.

[0191] Example 28

[0192] Referring to the synthesis of Example 5, in Step 2, N-Boc-6-aminohexanoic acid was used as a raw material for replacement, and in Step 3, cis-5,8,11,14,17-eicosapentaenoic acid was used as a raw material for replacement to synthesize the molecule of C20U5L5V (the structure is shown below).

[0193]

[0194] 1 1H NMR (600 MHz, CDCl3) δ 8.68 (s, 1H), 7.47 (t, J J = 6.0 Hz, 1H), 7.38 –7.32 (m, 4H), 6.41 (d, J J = 9.2 Hz, 1H), 6.01 (t, J J = 5.9 Hz, 1H), 5.43 – 5.28 (m,10H), 4.70 (t, J J = 8.0 Hz, 1H), 4.56 – 4.49 (m, 3H), 4.36 (dd, J= 15.1, 5.5 Hz, 1H), 4.01 (d, J = 11.2 Hz, 1H), 3.75 – 3.63 (m, 3H), 3.18 (p, J = 6.6 Hz, 4H), 2.87 – 2.76 (m, 8H), 2.50 (s, 3H), 2.43 – 2.37 (m, 1H), 2.25 – 2.11 (m, 6H), 2.08 (q, J = 6.8 Hz, 4H), 1.66 (p, J = 7.5 Hz, 2H), 1.59 (p, J = 7.6 Hz, 2H), 1.33– 1.24 (m, 2H), 1.02 – 0.92 (m, 12H). 13 C NMR (151 MHz, CDCl3) δ 173.56, 173.28, 171.67, 171.19, 150.37, 148.36, 138.23, 132.05, 131.66, 130.80, 129.44, 129.11, 128.69, 128.58, 128.28, 128.19, 128.18, 128.05, 127.98, 127.84, 126.98, 70.00, 58.82, 57.57, 56.80, 55.51, 53.47, 43.47, 43.11, 39.27, 36.52, 36.09, 36.06, 35.21, 29.14, 26.68, 26.43, 26.21, 25.67, 25.62, 25.53, 25.12, 20.55, 18.52, 17.06, 16.05, 14.28, 12.67. ESI-MS (m / z): [M+K] calculated for C 48 H 68 N4O5S 850.4547, found: 850.488.

[0195] Example 29

[0196] CCK8 cell proliferation assay: PANC-1 and MIAPACA2 cells were incubated with different concentrations of the above compounds for 72 h. The CCK8 cell proliferation activity assay showed that C18U2L3V had the highest pancreatic cancer killing efficiency (see Figure 2-1 ), and by measuring the IC 50The area under the region was integrated and it was found that pancreatic cancer was highly sensitive to drugs of C20U4LV, C20U4L1V, C18U2L3V, and C18U2L4OV (see Figure 2-2 , Figure 2-3 ).

[0197] Embodiment 30

[0198] Combination of the two drugs: Cell proliferation assay-plate clone formation experiment was used to detect the effect of the combination of the two drugs on the proliferation ability of pancreatic tumor cells. Different concentrations of C20U4LV (1μm, 3μm, 5μm) and MOF-Fe@PEI (0.3μg / ml, 1μg / ml, 3 μg / ml) were used alone and in combination. It was found that when the two drugs were used in combination, they had a strong synergistic effect (see Figure 3 In the human pancreatic cancer cell line PANC-1, compared with the control group without drug addition, the cell clone percentage after combined treatment with 1 μm C20U4LV and 3 μg / ml MOF-Fe@PEI was 10.3%, which was much lower than the single application of 1 μm C20U4LV (cell clone percentage: 89.13%) and the single application of 3 μg / ml MOF-Fe@PEI (cell clone percentage: 32.6%).

[0199] Embodiment 31

[0200] Western blotting (WB) experiment: Based on the experimental results of Example 29, the cell killing effect and drug sensitivity of the above compounds on PANC-1 and MIAPACA2 cells were comprehensively considered, and four compounds (C20U4LV, C20U4L1V, C18U2L3V, C18U2L4OV) were selected for ferritin degradation experimental verification. Due to the use of MOF-Fe@PROTAC@PEI dosage form, MOF-Fe@PEI (10μg / ml) was added at the same time as the drug (10μm / ml), and WB experiment was performed after incubation with PANC-1 cells for 48h. Western blotting was used to analyze protein, cells were lysed using lysis buffer, and protein concentration was determined by BCA kit. 20μg of protein was taken for immunoblotting analysis, and FTH1 (1:3000, (A1144), ABclonal) and β-Actin (1:4000, (AC026), ABclonal) were used to characterize the degradation efficiency of ferritin. The experimental results show that MOF-Fe can significantly upregulate the expression of ferritin by delivering iron ions into pancreatic cancer, while C18U2L3V and C20U4LV can effectively degrade the expression of ferritin (see Figure 4 ).

[0201] Embodiment 32

[0202] On the other hand, in order to further prove the mechanism of this type of PROTAC molecule, the VHL ligand drug ((2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride) was used to inhibit the performance of PROTAC (taking C20U4LV as an example). Western blotting (WB) experiments showed that the addition of VHL-ligand could effectively inhibit the degradation of ferritin by this molecule, further confirming that this type of PROTAC molecule binds through the E3 ubiquitin ligase, highly ubiquitinates ferritin, and finally degrades through the proteasome pathway (see Figure 5 ).

[0203] Example 33

[0204] Effect of MOF-Fe@PEI drug-loaded PROTAC to obtain the compound of general formula I: In order to better analyze the anti-cancer efficiency of the compound of general formula I in vivo and overcome the drug disadvantages of PROTAC molecules, through the use of the MOF-Fe@PEI drug-loaded delivery strategy, the MOF-Fe@PEI nanodrug improved the targeting and drug delivery efficiency of PROTAC small molecules, and the molecule was observed to have good stability by electron microscopy (as Figure 6 shown, taking MOF-Fe@PEI@C20U4LV as an example). The preparation of MOF-Fe@PEI drug-loaded PROTAC to obtain the compound of general formula I includes the following steps: Add 2-aminoterephthalic acid (NH2-BDC) (200 mg) and iron(III) chloride hexahydrate (FeCl3·6H2O) (600 mg) to the reaction kettle, and react at 110 °C for 48 h. After drying in an oven at 100 °C, take 100 mg of MOF-Fe and add PROTAC (10 mg), ultrasonically disperse, and incubate at room temperature for 24 h. Centrifuge and wash the above nanoparticles, add cationic polyimine and incubate for 24 h, centrifuge to collect the precipitate, and resuspend.

[0205] Example 34

[0206] Term definitions and explanations:

[0207] Unless otherwise specified, the group and term definitions recorded in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be combined and combined with each other arbitrarily. The group definitions and compound structures after such combination and combination shall fall within the scope recorded in the specification of this application.

[0208] Unless otherwise specified, the numerical ranges recited in this specification and the claims are equivalent to at least reciting each specific integer value therein. For example, the numerical range "1-10" is equivalent to reciting each integer value in the numerical range "1-10", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10. It should be understood that in the description of substituents herein, for "one, two or more", "more" should refer to an integer ≥3, such as 3, 4, 5, 6, 7, 8, 9 or 10.

[0209] The term "C 1-50 alkyl chain" means a straight-chain or branched-chain alkyl having 1-50 carbon atoms.

[0210] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A ferroptosis-inducing nanodrug targeting ferritin, characterized in that, Containing a MOF-Fe@PROTAC@PEI complex, the MOF-Fe@PROTAC@PEI complex comprises a compound and MOF-Fe nanoparticles. The MOF-Fe nanoparticles load the compound through the internal pore diameter to form MOF-Fe@PROTAC, and the surface of the MOF-Fe@PROTAC is wrapped by cationic polyimine to form the MOF-Fe@PROTAC@PEI complex; The cationic polyimine is polyetherimide PEI; The compound is selected from four categories of C18U1L3V, C20U4LV, C20U4L1V, and C18U2L3V, and the structural formulas are respectively: 。 2. Preparation method of ferritin-targeted ferroptosis-inducing nanodrug delivery system, characterized in that, Including: Step 1, prepare the compound; Step 2, prepare MOF-Fe nanoparticles; Step 3, prepare the MOF-Fe@PROTAC@PEI complex. Load the MOF-Fe nanoparticles prepared in Step 2 with the compound prepared in Step 1 through the internal pore diameter to form MOF-Fe@PROTAC, and then wrap the surface of the MOF-Fe@PROTAC with polyetherimide PEI to form the MOF-Fe@PROTAC@PEI complex, and further prepare a ferroptosis inducer-targeted ferritin ferroptosis-inducing nanodose form; The compound is selected from four categories of C18U1L3V, C20U4LV, C20U4L1V, and C18U2L3V, and the structural formulas are respectively: The synthesis steps of C20U4LV are as follows: Dissolve (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride, which recruits E3 ubiquitin ligase, and arachidonic acid in organic reagent A, add the condensation reagent HATU and the co-used base solution DIPEA, protect with nitrogen, react at room temperature overnight, extract with 5% sodium carbonate solution and organic reagent B to obtain the crude reaction product, and purify by silica gel column to obtain a pale yellow solid; The synthesis steps of C20U4L1V are as follows: Step 1.1: Synthesis of compound 1. Dissolve (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride, which recruits E3 ubiquitin ligase, and arachidonic acid in organic reagent A, add the condensation reagent HATU and the co-used base solution DIPEA, protect with nitrogen, react at room temperature overnight, extract with 5% sodium carbonate solution and organic reagent B to obtain the crude reaction product, and purify by silica gel column to obtain a pale yellow solid; Step 1.2: Synthesis of compound 2. Dissolve the compound 1 in dichloromethane, add hydrochloric acid-1,4-dioxane, react at room temperature for 30 minutes, and obtain the crude product after rotary evaporation; Step 1.3: Synthesis of compound C20U4L1V. Dissolve the said compound 2 and arachidonic acid in organic reagent A, add condensation reagent HATU and the co-used base solution DIPEA, protect with nitrogen, react overnight at room temperature, extract the crude reaction product with 5% sodium carbonate solution and organic reagent B, and purify by silica gel column to obtain a white solid; The synthesis steps of C18U1L3V are as follows: Refer to the synthesis steps of C20U4L1V, use N-Boc-γ-aminobutyric acid as the raw material, replace arachidonic acid used in the synthesis of compound 1 in Step 1.1 of the C20U4L1V synthesis, use oleic acid as the raw material, replace arachidonic acid used in Step 1.3 of the C20U4L1V synthesis, and synthesize to obtain white solid C18U1L3V; The synthesis steps of C18U2L3V are as follows: Refer to the synthesis steps of C20U4L1V, use N-Boc-γ-aminobutyric acid as the raw material, replace arachidonic acid used in the synthesis of compound 1 in Step 1.1 of the C20U4L1V synthesis, use linoleic acid as the raw material, replace arachidonic acid used in Step 1.3 of the C20U4L1V synthesis, and synthesize to obtain white solid C18U2L3V.

3. The preparation method of the ferroptosis-inducing nanodrug formulation targeting ferritin according to claim 2, characterized in that, The said organic reagent A is selected from dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, dichloromethane; the said organic reagent B is selected from ethyl acetate, dichloromethane.

4. The preparation method of the ferroptosis-inducing nanodrug formulation targeting ferritin according to claim 2, wherein, The preparation steps of the MOF-Fe@PROTAC@PEI composite include the following steps: Step 21: Add 2-aminoterephthalic acid NH2-BDC and iron(III) chloride hexahydrate FeCl3·6H2O into a reaction kettle, react at 110 °C for 48 h; Step 22: After drying in an oven at 100 °C, add the compound, disperse by ultrasonic wave, and incubate at room temperature for 24 h; Step 23: Centrifuge and wash the above nanoparticles, add polyetherimide PEI and incubate for 24 h, centrifuge to collect the precipitate, and resuspend.

5. Use of the compound or the MOF-Fe@PROTAC@PEI composite according to claim 1 in the preparation of a drug for treating pancreatic cancer.

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