Process for the preparation of lometabolic derivatives and their pharmaceutical activity

Abstract

This invention belongs to the fields of organic chemistry, medicinal chemistry, and pharmacology, specifically relating to the synthesis of lometapeptide bioelectron isosteres and their application in the preparation of therapeutic drugs for lipid-lowering diseases. A method for preparing lometapeptide bioelectron isosteres mainly includes six steps: 1) Using A and B as starting materials, under nickel catalysis and the action of phosphine ligands, a reaction occurs... endo The reaction [2+2] yielded intermediate I; 2) Intermediate I underwent intramolecular cyclization under thioxanthracene-9-one and photocatalysis to yield intermediate II; 3) Intermediate II was oxidized under alkaline potassium permanganate to yield intermediate III; 4) Intermediate III and C reacted under the action of a condensing agent to prepare amide intermediate IV; 5) Intermediate IV was debenzylated under the action of palladium on carbon and a hydrogen source to prepare amine intermediate V; 6) Intermediate V and D under alkaline conditions underwent a substitution reaction to obtain the final product, lometapetine bioelectron isostere VI. This bioelectron isostere can dose-dependently inhibit the secretion of apolipoprotein B in HepG2 cells and exhibits good kinetic solubility. This compound shows promise for development as a therapeutic drug for hypercholesterolemia-related diseases.

Description

[0001] This invention relates to the fields of drug synthesis and pharmacology, specifically to a method for preparing and applying a lometabolic bioelectron isostere. This compound achieves a lipid-lowering effect by inhibiting the release of apolipoproteins (such as apoB) from liver cells, and has the potential to be developed into a drug for the treatment of hypercholesterolemia. Background Technology

[0002] Ortho-disubstituted benzene ring skeletons are widely found in drug molecules and bioactive substances [Black, HR; Bailey, J.; Zappe, D., et al., Valsartan: more than a decade of experience]. Drugs 2009, 69 , 2393-2414. Sharpe, M.; Jarvis, B.; Goa, KL, Telmisartan: review of its use in hypertension. Drugs 2001, 61[1501-1529.] (Figure 1), for example, commercially available aspirin and drug molecules such as Valsartanz and Telmisartan used to treat hypertension, as well as the well-known fungicides Fluxapyroxade and Boscalid in agrochemistry, all contain ortho-disubstituted benzene ring structural units. Lomitapide is a drug used to treat familial hypercholesterolemia. Its main action is to reduce LDL cholesterol levels in the blood by inhibiting the synthesis of low-density lipoprotein (LDL) in the liver. This drug has shown significant effects in managing some difficult-to-control cases of high cholesterol. In addition to directly affecting LDL cholesterol, lomitapide can also reduce total cholesterol and non-high-density lipoprotein (non-HDL) cholesterol levels. This is crucial for the comprehensive control of lipid metabolism in patients, as non-HDL cholesterol is closely related to the development of atherosclerosis and cardiovascular disease. Lomitapide helps to improve the overall balance of lipid metabolism by regulating the lipid metabolism process in the liver, not only by reducing the level of specific types of cholesterol. This positive effect can manifest as a more stable and lasting effect in long-term treatment, helping to prevent and reduce the risk of cardiovascular disease. In recent years, replacing the benzene ring with saturated bioisosteres has become an important strategy for obtaining novel patent-free molecules with superior bioactivity and physicochemical properties. However, these synthetic methods for the skeleton of ortho-substituted benzene saturated isosteres typically require the pre-synthesis of mono- or para-substituted saturated isosteres [Denisenko, A.; Garbuz, P.; Shishkina, SV, et al., Saturated Bioisosteres of ortho-Substituted Benzenes]. Angew. Chem. Int. Ed 2020, 59 ,20515-20521. Harmata, AS; Spiller, TE; Sowden, MJ, et al.,Photochemical Formal (4+2)-Cycloaddition of Imine-Substituted Bicyclo[1.1.1]pentanes and Alkenes. J. Am. Chem.Soc 2021, 143[21223-21228.], while also having problems such as complex synthetic routes, stringent reaction conditions, and low overall yield [Lopchuk, JM; Fjelbye, K.; Kawamata, Y., et al., Strain-Release Heteroatom Functionalization: Development, Scope, and Stereospecificity. J. Am. Chem. Soc 2017, 139 [3209-3226.]. Therefore, developing a new, efficient, and simple method for isosteric bioelectrons of ortho-substituted benzene rings is of great significance.

[0003]

[0004] Formula 1: Bioactive molecules with ortho-disubstituted benzene ring structures Baran describes [Zhao, JX; Chang, YX; He, C., et al., 1,2-Difunctionalized bicyclo[1.1.1] pentanes: Long-sought-after mimetics forortho / meta-substituted arenes. Proc. Natl. Acad. Sci. U.S.A 2021, 118 [e2108881118.] A multifunctional platform for the synthesis of 1,2-difunctionalized bicyclo[1.1.1]pentane has been developed to potentially mimic ortho / meta-substituted aromatics. Through a systematic analysis of key properties of several drugs and their bioequivalence analogs, specifically, an increase in solubility is typically observed with such substitution (Equation 2).

[0005]

[0006] High-cubic alkylene exhibits properties similar to cubic alkylene [Hasegawa, T.; Kuwatani, Y.; Higuchi, H., et al., Wagner-Meerwein Rearrangement of 4-Hydroxymethylpentacyclo[4.3.0.02,5.03,8.04,7]nonane to Pentacyclo[5.3.0.02,6.03,9.04,8]decane inFormic Acid. Bull. Chem. Soc. Jpn 1993,66 [3009-3014.], these polyhedral alkanes are rigid and have high CH bond energies due to their strain characteristics. Compared to bicyclic [2.1.1]hexanes, polyhedral alkanes better match the benzene ring structure. Existing literature reports schemes for constructing bioisosteres of disubstituted benzene rings using high-cubic alkanes [Takebe, H.; Matsubara, S., Scaffold Editing of Cubanes into Homocubanes, Homocuneanes via Cuneanes.] Chem-Eur J 2024, 30 [e202303063.]. However, there are no literature reports on ortho-disubstituted bioisosteres.

[0007] Based on this, the present invention aims to develop a method for preparing lometabolic bioelectron isosteres and their application in the preparation of therapeutic drugs for lipid-lowering drug-related diseases.

[0008] Formula 2 Lomitapide and its isosteres (+)-E and (-)-E Summary of the Invention

[0009] The purpose of this invention is to provide a method for preparing lometapeptide bioelectron isosteres. This method has a short procedure, simple and easy reaction, is applicable to gram-scale reactions, and can be used for the large-scale synthesis of lometapeptide bioelectron isosteres.

[0010] The technical solution of this invention is: a method for preparing lometasone bioelectron isosteres, the specific synthetic route of which is as follows:

[0011] The specific synthesis steps are as follows: 1) Preparation of intermediate I: Using A and B as starting materials, the reaction proceeds under nickel catalysis and the action of phosphine ligands. endo The [2+2] reaction was used to prepare intermediate I; 2) Preparation of intermediate II: Intermediate I undergoes intramolecular cyclization under the action of a photocatalyst to obtain intermediate II; 3) Preparation of intermediate III: Intermediate II was oxidized under the action of an oxidant to prepare intermediate III; 4) Preparation of intermediate IV: Intermediate III and C were reacted under the action of a condensing agent to prepare amide compound intermediate IV; 5) Preparation of intermediate V: Intermediate IV was debenzylated under the action of palladium on carbon and hydrogen source to prepare amine compound intermediate V; 6) Preparation of bioelectron isostere VI: Intermediates V and D undergo a substitution reaction under alkaline conditions to obtain the final product VI.

[0012] In step 1, the range of phosphine ligands selected includes, but is not limited to, triphenylphosphine, tri-tert-butylphosphine, 1,4-bis(diphenylphosphine)butane, bis(2-diphenylphosphine) ether, 1,1'-binaphthyl-2,2'-bisdiphenylphosphine, tris(4-methoxyphenyl)phosphine, tris(pentafluorophenyl)phosphine, tris(4-trifluorotolyl)phosphine, triethyl phosphite, tris(4-fluorophenyl)phosphine, tris[3,5-di(trifluoromethyl)phenyl]phosphine, etc., among which tris[3,5-di(trifluoromethyl)phenyl]phosphine is the best.

[0013] In step 1, the solvent selection range includes, but is not limited to, tetrahydrofuran, dimethyl sulfoxide, N,N-dimethylformamide, ethylene glycol dimethyl ether, n-hexane, acetonitrile, m-xylene, p-xylene, cyclohexane, and toluene, with cyclohexane being the most suitable. The nickel catalyst is bis-(1,5-cyclooctadiene)nickel, and the molar ratio of A, B, nickel catalyst, and phosphine ligand is 1.0:2.0:0.1:0.2.

[0014] In step 2, the photocatalyst can be selected from a range including, but not limited to, thioxanthracene-9-one derivatives and 9-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)phenyl]-carbazole derivatives, with 3-methoxy-thioxanthracene-9-one being the most suitable. The solvent can be selected from a range including, but not limited to, tetrahydrofuran, dimethyl sulfoxide, diethyl ether, ethylene glycol dimethyl ether, n-hexane, acetonitrile, acetone, m-xylene, p-xylene, and toluene, with toluene being the most suitable. The molar ratio of intermediate I to the photocatalyst is 1.0:0.1.

[0015] In step 3, the oxidant can be selected from a range including, but not limited to, potassium permanganate, Jones reagent, and hydrated ferric nitrate, with potassium permanganate being the best. The molar ratio of intermediate II to oxidant is 1.0:4.0.

[0016] In step 4, the selection range of condensing agents includes, but is not limited to, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide condensing agents, and onium salt condensing agents, among which 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride is the best. The condensation aid is 1-hydroxybenzotriazole. The molar ratio of intermediates III and C, the condensing agent, and the condensation aid is 1.0:1.2:1.2:1.4.

[0017] In step 5, the hydrogen source can be selected from, but is not limited to, formic acid, ammonium formate, hydrogen, and water, with formic acid being the best. The molar ratio of intermediate IV to formic acid is 1.0:5.0.

[0018] In step 6, the range of alkalis to be selected includes, but is not limited to, triethylamine, potassium carbonate, sodium hydride, and sodium hydroxide, with triethylamine being the best. The molar ratio of intermediates V, D, and alkali is 1.0:1.2:3.0.

[0019] The beneficial effects of this invention are: it provides a method for preparing lometapeptide bioelectron isosteres, which has a short route and simple and easy reaction, is applicable to gram-scale reactions, and can be used for the large-scale synthesis of lometapeptide bioelectron isosteres.

[0020] Furthermore, the present invention excludes endo / exo The possibility of interconversion of products due to configurational inversion in the reaction system. This invention realizes the reaction of norbornene with unactivated alkynes catalyzed by zero-valent nickel. endo The [2+2] type reaction exhibits good yields. endo Excellent selectivity. Meanwhile, focusing on the research of photocatalytic intramolecular olefin [2+2] cyclization synthesis of ortho-disubstituted high cuboidal alkanes, commercially available norbornene and alkynes can be used as starting materials to rapidly construct ortho-disubstituted high cuboidal alkanes and apply them to the synthesis of lometasyl isosteres.

[0021] To test the application of lometape bioelectron isosteres in lipid-lowering drugs, this patent uses HepG2 cells as a carrier to test the effects of lometape and lometape bioelectron isosteres on apolipoprotein (apoB) in HepG2 cells (Figure 1). In the presence of oleic acid and palmitic acid (500 μM / 250 μM), HepG2 cells were treated with different concentrations of lometape and lometape bioelectron isosteres for 24 h. The accumulation of apolipoprotein B in the culture medium was measured by ELISA. The data are expressed as mean ± SEM (n=3). The half-maximal inhibitory concentrations (IC50) of lometape (IC50=113.3 nM) and lometape bioelectron isosteres (IC50=33.2 nM) were calculated. To evaluate the solubility and precipitation behavior of compounds in aqueous solutions, providing crucial decision-making support for compound screening, optimization, and biological experimental design, and significantly reducing late-stage development risks and costs, this patent tested the kinetic solubility of lometabine and its bioelectron isostere (19.7 μg / mL vs. 258.6 μg / mL), showing a more than tenfold increase in solubility (Figure 2). Simultaneously, to rapidly assess the new drug potential of the lometabine bioelectron isostere, this patent conducted mouse liver microsomal metabolic stability analysis, testing its phase metabolic rate and predicting its in vivo clearance potential (Figure 3). Detailed Implementation

[0022] The following embodiments are further illustrations of the present invention, and not intended to limit the scope of the invention. Modifications and substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit of the invention are all within the scope of the invention.

[0023] The following embodiments are further illustrations of the present invention, and not intended to limit the scope of the invention. Modifications and substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit of the invention are all within the scope of the invention. Example

[0024] Step 1: Preparation of Intermediate I:

[0025] Inside the glove box, add A (0.2 mmol, 40.0 mg), norbornene B (0.4 mmol, 36.9 mg), bis(1,5-cyclooctadiene)nickel (0.02 mmol, 5.5 mg), tris(3,5-bis(trifluoromethylphenyl)phosphine) (0.04 mmol, 26.8 mg), and toluene (0.5 mL) to a sealed tube. Seal the tube and remove it from the glove box. Incubate at 80°C. o The mixture was stirred in reaction module C for 12 hours. After the reaction was complete, it was filtered through diatomaceous earth, concentrated, dissolved in deuterated chloroform, and then sampled for 1H NMR analysis. The results were recorded. endo / exo After adjusting the ratio of the two isomers, the crude product was mixed with NMR recovery solution, the solvent was evaporated under reduced pressure, and column chromatography was performed to give yellow oil I (37.0 mg), yield 63%, with an isomer ratio of [missing information]. endo : exo = 83:17. The following are the NMR experimental data for product I: 1 H NMR (500 MHz, CDCl3) δ= 7.54 (d, J = 8.0 Hz, 2H), 7.33 (d, J = 8.0Hz, 2H), 5.93-5.69 (m, 2H), 4.38-4.28 (m, 2H), 3.19-3.14 (m, 1H), 3.08-3.03(m, 1H), 2.91 (s, 1H), 2.79 (s, 1H), 1.96 (d, J = 8.0 Hz, 1H), 1.64 (d, J =8.0 Hz, 1H). 13C NMR (125 MHz, CDCl3) δ= 147.5, 142.2, 137.9, 132.1, 131.6, 128.8(q, J C-F = 32.4 Hz), 127.3, 126.2, 125.3 (q, J C-F = 3.75 Hz), 124.1 (q, J C-F =262.9 Hz), 59.6, 53.7, 42.01, 41.96, 41.0. 19 F NMR (470 MHz, CDCl3) δ = -62.4. Step 2, Preparation of Intermediate II:

[0026] In a 25 mL Schlenk tube equipped with a magnetic stir bar, intermediate II (210 mg, 0.72 mmol) and a solution of potassium permanganate (454 mg, 2.9 mmol) in H₂O (5% KOH, 5 mL) were added at room temperature. The mixture was slowly heated to 25 °C and stirred overnight. The mixture was treated with HCl (2 M) until acidic, and then extracted with dichloromethane (20 mL × 3). The bound organic phase was washed with saturated NaCl solution, dried over anhydrous Na₂SO₄, and concentrated under vacuum. The residue was then purified by column chromatography on silica gel (ethyl acetate / petroleum ether = 1:3) to give 210 mg (95%) of product III as a white solid. The following are the NMR experimental data of product III: 1 H NMR (500 MHz, CDCl3) δ=7.55 (d, J = 8.0 Hz, 2H), 7.29 (d, J = 8.0Hz, 2H), 3.59-3.48 (m, 4H), 3.47-3.43 (m, 1H), 3.37-3.32 (m, 1H), 1.80 (s, 2H). 13 C NMR (125 MHz, CDCl3) δ= 176.6, 142.9, 128.5 (q, J C-F = 32.0 Hz), 126.4, 125.1 (q, J C-F= 3.6 Hz), 124.3 (q, J C-F = 270.4 Hz), 55.6, 45.8, 44.0,43.7, 42.7, 42.0. 19 F NMR (470 MHz, CDCl3) δ = -62.4. Step 3: Preparation of Intermediate III:

[0027] In a 25 mL Schlenk tube equipped with a magnetic stir bar, intermediate II (210 mg, 0.72 mmol) and a solution of potassium permanganate (454 mg, 2.9 mmol) in H₂O (5% KOH, 5 mL) were added at room temperature. The mixture was slowly heated to 25 °C and stirred overnight. The mixture was treated with HCl (2 M) until acidic, and then extracted with dichloromethane (20 mL × 3). The bound organic phase was washed with saturated NaCl solution, dried over anhydrous Na₂SO₄, and concentrated under vacuum. The residue was then purified by column chromatography on silica gel (ethyl acetate / petroleum ether = 1:3) to give 210 mg (95%) of product III as a white solid. The following are the NMR experimental data of product III: 1 H NMR (500 MHz, CDCl3) δ=7.55 (d, J = 8.0 Hz, 2H), 7.29 (d, J = 8.0Hz, 2H), 3.59-3.48 (m, 4H), 3.47-3.43 (m, 1H), 3.37-3.32 (m, 1H), 1.80 (s, 2H). 13 C NMR (125 MHz, CDCl3) δ= 176.6, 142.9, 128.5 (q, J C-F = 32.0 Hz), 126.4, 125.1 (q, J C-F = 3.6 Hz), 124.3 (q, J C-F = 270.4 Hz), 55.6, 45.8, 44.0,43.7, 42.7, 42.0. 19 F NMR (470 MHz, CDCl3) δ = -62.4. Step 4: Preparation of Intermediate IV:

[0028] Intermediate III (8.0 mmol, 2.45 g), C (9.6 mmol, 1.83 g), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (9.6 mmol, 1.49 g), 1-hydroxybenzotriazole (11.2 mmol, 1.51 g), and dimethylformamide (25 mL) were added to a 25 mL oven-dried Schlenk tube fitted with a magnetic stir bar. The resulting mixture was stirred overnight at the same temperature. After the reaction was complete, it was quenched with saturated sodium bicarbonate. The mixture was extracted with dichloromethane (50 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, concentrated under vacuum, and purified by flash column chromatography on silica gel (ethyl acetate / petroleum ether = 1:5) to give 3.56 g (96%) of the desired product IV as a white solid. The following are the NMR experimental data of product IV: 1 H NMR (500 MHz, CDCl3) δ= 7.59 (d, J = 8.0 Hz, 2H), 7.34-7.21 (m,7H), 4.89 (d, J = 8.0 Hz, 1H), 3.72-3.63 (m, 1H), 3.53-3.48 (m, 4H), 3.47-3.44 (m, 1H), 3.40 (s, 2H), 3.36-3.32 (m, 1H), 2.61-2.44 (m, 2H), 2.04 (t, J = 10.5 Hz, 2H), 1.80 (s, 2H), 1.69-1.61 (m, 2H), 1.23-1.12 (m, 2H). 13 C NMR (125 MHz, CDCl3) δ= 169.5, 143.3, 138.1, 129.0, 128.6 (q, J C-F = 32.3 Hz), 128.1, 127.0, 126.4, 125.4 (q, J C-F = 3.5 Hz), 124.1 (q, J C-F=270.3 Hz), 62.90 (s), 55.34, 55.29, 51.7, 45.5, 44.2, 43.7, 42.5, 41.9, 31.8. 19 F NMR (470 MHz, CDCl3) δ = -62.3. Step 5: Preparation of intermediate V:

[0029] A solution of intermediate IV (1.91 g, 4.0 mmol), formic acid (0.92 g, 20.0 mmol), and 20% Pd-C (0.85 g) in methanol (20 mL) was stirred overnight at 70 °C. The catalyst was filtered through diatomaceous earth and washed with methanol (10 mL). The filtrate was concentrated under vacuum and purified by flash column chromatography on silica gel (methanol / dichloromethane = 1:20) to give 1.24 g (80%) of the desired product V as a white solid. The following are the NMR experimental data of product V: 1 H NMR (500 MHz, CDCl3) δ= 7.57 (d, J = 8.0 Hz, 2H), 7.30 (d, J = 8.0Hz, 2H), 5.51 (s, 1H), 3.87-3.78 (m, 1H), 3.56-3.49 (m, 4H), 3.46-3.34 (m,2H), 3.20 (d, J = 12.0 Hz, 2H), 2.77 (t, J = 11.5 Hz, 2H), 1.82 (s, 2H), 1.78(d, J = 13.0 Hz, 2H), 1.61-1.50 (m, 2H). 13 C NMR (125 MHz, CDCl3) δ= 169.7, 143.2, 128.4 (q, J C-F = 32.3 Hz), 126.3, 125.2 (q, J C-F = 3.6 Hz), 124.1 (q, J C-F = 270.3 Hz), 55.4, 45.5, 44.10, 44.07, 43.4, 43.0, 42.5, 41.9, 29.4. 19 F NMR (470 MHz, CDCl3) δ = -62.3. Step Six: Preparation of Bioelectron Isosterone VI:

[0030] Compound D (469 mg, 1.1 mmol), intermediate V (350 mg, 0.9 mmol), triethylamine (273 mg, 2.7 mmol), and acetonitrile (5 mL) were added to a 25 mL Schlenk tube fitted with a magnetic stir bar. The resulting mixture was stirred overnight at the same temperature. After the reaction was complete, the mixture was concentrated under vacuum and purified by flash column chromatography on silica gel (methanol / dichloromethane = 1:30) to give 600 mg (91%) of the desired product VI as a white solid. The following are the NMR experimental data of product VI: 1 H NMR (500 MHz, CDCl3) δ= 7.76 (d, J = 7.5 Hz, 2H), 7.58-7.50 (m,4H), 7.44 (t, J = 7.5 Hz, 2H), 7.36 (t, J = 7.5 Hz, 2H), 7.29 (d, J = 7.5 Hz, 2H), 5.38 (t, J = 6.0 Hz, 1H), 5.07 (d, J = 7.5 Hz, 1H), 3.73-3.63 (m, 3H), 3.52-3.47 (m, 4H), 3.46-3.41 (m, 1H), 3.37-3.32 (m, 1H), 2.68-2.55 (m, 1H), 2.44-2.37 (m, 2H), 2.21-2.14 (m, 2H), 2.06 (t, J = 11.0 Hz, 2H), 1.80 (s,2H), 1.67-1.59 (m, 2H), 1.41-1.30 (m, 4H), 0.72-0.63 (m, 2H). 13 C NMR (125 MHz, CDCl3) δ= 173.3, 169.7, 144.9, 143.1, 140.9, 128.6,128.4 (q, J C-F= 32.3 Hz), 128.1, 126.4, 125.3 (q, J C-F = 3.5 Hz), 124.2, 123.7(q, J C-F = 277.1 Hz), 120.4, 62.2, 57.7, 55.3 (d, J = 5.5 Hz), 51.6, 45.4,44.9, 44.1, 43.5, 42.5, 41.9, 40.6 (q, J C-F = 34.5 Hz), 35.8, 30.7, 26.0,21.5. 19 F NMR (470 MHz, CDCl3) δ = -62.3, -72.8 (t, J = 8.5 Hz). Example 2: Effects of lomestatide and lomestatide bioelectron isostere VI on apolipoprotein (apoB) in HepG2 cells First, cell viability was measured to rule out the influence of drug toxicity on cells. HepG2 cells were stored in DMEM supplemented with 10% bovine serum and 1% penicillin-streptomycin at 37°C under a 5% CO2 atmosphere. Cells were seeded at a density of 5 × 10^3 cells per well in 96-well plates and allowed to grow to ~80% confluence before treatment with the specified compound for 24 hours. Subsequently, 10 μL of CCK-8 reagent (Mei5 Biotechnology, MF128-01) was added to each well, and the cells were incubated at 37°C for 2 hours. The absorbance at 450 nm was measured using a SpectraMax iD5 microplate reader.

[0031] Cells were cultured in Minimum Essential Medium (MEM; Meilunbio®) containing 10% fetal bovine serum (FBS; ExCell) and 1% penicillin-streptomycin mixture at 37°C and 5% CO2 atmosphere. Cells were sputtered at a rate of 2 × 10⁶ cells / year. 5Cells were seeded at a density of cells / well in 6-well plates and cultured for 24 hours. Except for the control group, oleic acid and palmitic acid (final concentrations of 500 μM and 250 μM, respectively) were added to the experimental cells, and incubation continued for 48 hours. Subsequently, the experimental cells were treated with lometabine and lometabine bioelectron isostere VI at final concentrations of 0.03, 0.1, 0.3, and 1 μM, respectively, and incubated for another 24 hours. Control cells (containing no oleic acid / palmitic acid and any drugs) and model cells (containing oleic acid / palmitic acid but no drugs) were treated with 0.1% DMSO (final concentration of the medium) as the solvent control. Conditioned culture media were collected according to the kit instructions, and apoB content was detected using the Human apoB Elisa kit reagent (Elabscience). Data analysis was performed using GraphPad Prism 8.2.1 software, and the IC50 value was determined by nonlinear regression analysis of the concentration-response data (Figure 1).

[0032] Example 3: Testing the kinetic solubility of lomestatide and its bioelectron isosteres The compound was dissolved in DMSO to prepare a separate stock solution (30 mg / mL). 1.0 mL of ultrapure water was added to a 1.5 mL Eppendorf tube, and then the stock solution was gradually added under gentle vortexing until the solution was saturated. The suspension was centrifuged at 3000 g for 15 min at room temperature. The supernatant was collected and immediately filtered through a 0.45 µm nylon membrane. To avoid membrane equilibrium effects, the initial 200 µL of filtrate was discarded. The filtrate was then diluted 1:1 (v / v) with acetonitrile, and aliquots were injected into the chromatographic system under validation conditions. The peak area was quantified according to the calibration curve, and the kinetic solubility was the measured concentration in the filtered aqueous phase after equilibrium (Figure 2).

[0033] Example 4: Metabolic stability analysis of lomestatide and lomestatide bioelectron isosteres in mouse liver microsomes The test compound was incubated with mouse liver microsomes in the presence of NADPH. The reaction mixture consisted of NADPH (1 mM), liver microsomes (0.5 mg / mL), PBS buffer (100 mM, pH 7.4), and the test compound (1 μM). The reaction mixture was pre-incubated for 5 minutes, followed by the addition of the test compound to initiate the reaction. At 0, 10, 30, and 60 minutes of incubation, 50 μL of the reaction solution was transferred from the incubation plate to a sample plate containing 200 μL of cold acetonitrile (containing 50 ng / mL dexamethasone and toluene). A negative control (NC) sample was included, which did not contain the NADPH S114 regeneration system and was incubated with the compound and liver microsomes for 60 minutes. All samples were centrifuged at 12000 g for 5 minutes, and the supernatant was collected for LC-MS / MS analysis. Half-life (T0) was measured. 1 / 2 The formulas for calculating the remaining percentage of the compound are as follows: The slope k of the straight line is calculated using Microsoft Excel software (with the natural logarithm of the remaining percentage of the compound, ln(%), as the ordinate and the incubation time as the abscissa); where k (min) - ¹)=-slope, T 1 / 2 (min) = 0.693 / k, Remaining percentage of compound (% Remaining) = 100 × T / T0 (T is the concentration of compound detected at each time point, T0 is the initial concentration of compound at 0 minutes) (Appendix) Figure 3 ). Attached Figure Description

[0034] Figure 1 shows the effect of lometabine and lometabine bioelectron isosteres on apolipoprotein (apoB) in HepG2 cells.

[0035] Figure 2 It is used to test the kinetic solubility of lometasate and its bioelectron isosteres.

[0036] Figure 3 The purpose is to test the metabolic stability of lometaxel and lometaxel bioelectron isosteres in mouse liver microsomes.

[0037] Figure 4 It is the structural formula of the Lometas bioelectron isostere.

Claims

1. A method for synthesizing lometasate bioelectron isosteres and its application in the preparation of therapeutic drugs for lipid-lowering drug-related diseases. Characterized by, The specific synthetic route is as follows:

2. The method for preparing a lometasate bioisostere as described in claim 1, characterized in that, The specific synthesis steps are as follows: 1) Preparation of intermediate I: Using A and B as starting materials, the reaction proceeds under nickel catalysis and the action of phosphine ligands. endo The [2+2] reaction was used to prepare intermediate I; 2) Preparation of intermediate II: Intermediate I undergoes intramolecular cyclization under the action of a photocatalyst to obtain intermediate II; 3) Preparation of intermediate III: Intermediate II was oxidized under the action of an oxidant to prepare intermediate III; 4) Preparation of intermediate IV: Intermediate III and C were reacted under the action of a condensing agent to prepare amide compound intermediate IV; 5) Preparation of intermediate V: Intermediate IV was debenzylated under the action of palladium on carbon and hydrogen source to prepare amine compound intermediate V; 6) Preparation of bioelectron isostere VI: Intermediates V and D undergo a substitution reaction under alkaline conditions to obtain the final product VI.

3. The method for preparing a lometasate bioisostere according to claim 2, characterized in that, In step 1: the phosphine ligands include, but are not limited to, triphenylphosphine, tri-tert-butylphosphine, 1,4-bis(diphenylphosphine)butane, bis(2-diphenylphosphine) ether, 1,1'-binaphthyl-2,2'-bisdiphenylphosphine, tris(4-methoxyphenyl)phosphine, tris(pentafluorophenyl)phosphine, tris(4-trifluorotolyl)phosphine, triethyl phosphite, tris(4-fluorophenyl)phosphine, tris[3,5-bis(trifluoromethyl)phenyl]phosphine, etc.

4. The method for preparing a lometasate bioisostere according to claim 2, characterized in that, In step 1, the solvents include, but are not limited to, tetrahydrofuran, dimethyl sulfoxide, N,N-dimethylformamide, ethylene glycol dimethyl ether, n-hexane, acetonitrile, m-xylene, p-xylene, cyclohexane, and toluene.

5. The method for preparing a lometasate bioisostere according to claim 2, characterized in that, In step 2: the photocatalyst includes, but is not limited to, thioxanthracene-9-one derivatives, 9-[4-(4,6-diphenyl-1,3,5-triazin-2-yl)phenyl]-carbazole derivatives, etc.

6. The method for preparing a lometasate bioisostere according to claim 2, characterized in that, In step 2, the solvents include, but are not limited to, tetrahydrofuran, dimethyl sulfoxide, diethyl ether, ethylene glycol dimethyl ether, n-hexane, acetonitrile, acetone, m-xylene, p-xylene, and toluene.

7. The method for preparing a lometasate bioisostere according to claim 2, characterized in that, In step 3: the oxidizing agents include, but are not limited to, potassium permanganate, Jones reagent, and hydrated ferric nitrate.

8. The application according to claim 1, wherein the cause of the disease is at least in part caused by high cholesterol and high blood lipids.

9. The application according to claim 1, wherein the lipid-lowering effect is achieved by inhibiting the release of apolipoproteins (such as apoB) from liver cells, etc.

10. The application of the lometrepinephrine bioelectron isostere according to claim 1 in the treatment of various diseases at least partially caused by high cholesterol and high blood lipids, characterized in that, Including but not limited to the following diseases: atherosclerosis, coronary heart disease, stroke, peripheral vascular disease, kidney disease, fatty liver, and pancreatitis.

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