9-site demethylated anti-renal fibrosis natural product Commiphorane C-D, isomer thereof, and preparation method and application of 9-site demethylated anti-renal fibrosis natural product Commiphorane C-D

Abstract

The invention discloses a 9-site demethylated anti-renal fibrosis natural product Commiphorane C-D, an isomer thereof, and a preparation method and application of the 9-site demethylated anti-renal fibrosis natural product Commiphorane C-D, and belongs to the technical field of medicines. The invention specifically relates to compounds represented by formulas Ia, Ib, Ic and Id or isomers and pharmaceutically acceptable salts thereof, and a preparation method thereof. A new pharmaceutical composition comprises effective doses of compounds of formulae Ia, Ib, Ic and Id and a pharmacodynamically acceptable carrier. The invention also discloses application of the compound in prevention and / or treatment of renal fibrosis and / or chronic renal failure caused by renal fibrosis.

Description

Technical Field

[0001] This invention relates to a 9-demethylated natural product for treating renal fibrosis, Commiphorane CD, isomers and pharmaceutically acceptable salts thereof, their polycrystalline and eutectic forms, their precursors or derivatives / probes with the same biological function, their preparation methods, pharmaceutical compositions containing them, and their use as a medicine, particularly as a preventive / anti-renal fibrosis drug, belonging to the field of pharmaceutical technology. Background Technology

[0002] Renal fibrosis is a complex disease involving multiple pathogenic factors and mechanisms, and it is a significant factor in the progression of chronic kidney disease to renal failure. The etiology of renal fibrosis varies, and its initial location also differs, potentially starting in the glomeruli or interstitial region. However, the final outcome is the disappearance of normal glomerular and tubular structures, an increase in fibroblasts and myofibroblasts in the extracellular matrix, and may be accompanied by tubular atrophy, glomerular sclerosis, interstitial fibrosis, and progressive loss of renal function. Renal fibrosis involves multiple pathways, including inflammatory responses, apoptosis of intrinsic renal cells and immune cells, enhanced oxidative stress, and an imbalance of anti-fibrotic cytokines. Therefore, the development of anti-renal fibrosis drugs primarily focuses on inhibiting or blocking inflammation, anti-apoptosis, anti-oxidative stress, and regulating the local immune microenvironment of renal tissue. Although pirfenidone is currently used clinically to treat renal fibrosis, it has significant side effects. Therefore, developing new drugs with better activity and fewer toxic side effects is crucial for the current treatment of renal fibrosis.

[0003] In 2017, Cheng Yongxian's team extracted four terpenoid compounds with novel skeletons from Resina Commiphora, namely Commiphoranes AD. The article reported that the natural product Commiphorane C can inhibit the overexpression of fibronectin, collagen I, and α-smooth muscle actin in TGF-β1-induced proximal tubular cells of rat kidneys, exhibiting good anti-renal fibrosis activity. Naturally derived drugs are characterized by multiple targets and low toxicity; discovering new lead compounds and drugs from active natural drugs is currently one of the important directions in international drug research. The total synthesis of Commiphorane C has been reported, but the starting material is relatively expensive. Currently, the relationship between the structure and activity of Commiphorane C is unclear. Therefore, this invention proposes a simplified form of Commiphorane CD, a method for synthesizing isomers, and studies its pharmacological mechanism and potential therapeutic applications, especially for renal fibrosis and chronic renal failure, which has practical development value. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a 9-demethylated natural product, Commiphorane CD, isomer, pharmaceutically acceptable salt, prodrug, polycrystalline or cocrystalline form of which has excellent therapeutic effects and low toxicity and has activity in treating renal fibrosis and chronic renal failure.

[0005] Another technical problem to be solved by the present invention is to provide a method for preparing such compounds.

[0006] Another technical problem to be solved by the present invention is to provide a pharmaceutical composition containing such compounds.

[0007] Another technical problem to be solved by the present invention is to provide the use of such compounds in medicaments for treating renal fibrosis, chronic renal failure, and immune-related diseases.

[0008] To solve the technical problem of this invention, the following technical solution is adopted:

[0009] Compounds of formulas Ia, Ib, Ic and Id, or their isomers and pharmaceutically acceptable salts thereof;

[0010]

[0011] This invention also discloses a method for preparing the compounds of this invention, comprising the following steps:

[0012] Methods for preparing formulas Ia, Ib, Ic, and Id:

[0013]

[0014] i: Aldol reaction; ii: silylation reaction; iii: hydrolysis reaction; iv: witting reaction; v: borohydride oxidation; vi: reduction reaction; vii: substitution; viiii: cyclization reaction; ix: silylation / acetylation reaction; x: oxidation reaction; xi: hydrolysis / oxidation; xii: intramolecular Aldol reaction.

[0015] step:

[0016] i) 2-Bromo-3-benzyl-benzaldehyde 1 and EVANS reagent generate intermediate 3 via an asymmetric Aldol reaction. The reaction solvent is selected from dichloromethane, tetrahydrofuran, toluene, methanol, and dioxane. The reaction temperature is -80℃ to 25℃, and the reaction time is 0.5-10h.

[0017] ii) Intermediate 3 reacts with trimethylsilyl trifluoromethanesulfonate to obtain intermediate 4. The reaction solvent is selected from tetrahydrofuran or dichloromethane. The reaction temperature is -78℃ to 25℃ and the reaction time is 0.5 to 10 h.

[0018] iii) Intermediate 4 undergoes hydrolysis under organic or inorganic base conditions to generate compound 5; the organic base in the reaction is selected from: triethylamine, ethylenediamine, N,N-dimethylethylenediamine, propylenediamine, diisopropylaluminum hydride; the inorganic base is selected from: potassium carbonate, sodium carbonate, cesium carbonate, cesium fluoride; the reaction solvent is selected from: acetone, tetrahydrofuran, dichloromethane, toluene; the reaction temperature is -78℃ to 25℃, and the reaction time is 0.5 to 10 h;

[0019] iv) Compound 5 reacts with Witting's reagent under alkaline conditions to give compound 6; the base in the reaction is selected from: potassium carbonate, sodium carbonate, cesium carbonate, cesium fluoride, triethylamine, ethylenediamine, N,N-dimethylethylenediamine, propylenediamine, potassium tert-butoxide; the reaction solvent is selected from: dioxane, N,N-dimethylformamide, acetonitrile, water, ethylene glycol dimethyl ether, tetrahydrofuran, diethyl ether; the reaction temperature is -60℃ to 50℃, and the reaction time is 1 to 30 h;

[0020] v) Intermediate 6 was prepared into intermediate 7 by hydroboration oxidation; the reagents used were selected from: borane tetrahydrofuran complex, borane dimethyl sulfide complex, 9-BBN (9-boron bicyclo(3,3,1)-nonane), and the reaction solvents were selected from: dichloromethane, tetrahydrofuran, methanol, N,N-dimethylformamide, and water; the reaction temperature was -78℃ to 50℃, and the reaction time was 0.5 to 10 h;

[0021] vi) Intermediate 7 is reduced to obtain intermediate 8; the reducing agent used in the reaction is selected from: palladium on carbon, palladium hydroxide on carbon; the reaction solvent is selected from: dichloromethane, tetrahydrofuran, methanol, N,N-dimethylformamide; the reaction temperature is: -25℃-40℃, and the reaction time is 0.5-20h;

[0022] vii) Intermediate 8 undergoes an alkylation reaction with allyl bromide to generate intermediate 9; the base is selected from: sodium hydroxide, potassium carbonate, potassium hydroxide, triethylamine, ethylenediamine, pyridine; the reaction temperature is -25℃ to 40℃, and the reaction time is 0.5 to 20 h;

[0023] viii) Intermediate 9 undergoes an intramolecular cyclization reaction to transform into intermediate 10; the Pd catalyst is selected from: bis(dibenzylacetone)palladium, palladium acetate, 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride; the reaction solvent is selected from: dichloromethane, tetrahydrofuran, methanol, N,N-dimethylformamide, dioxane, toluene, and water; the reaction temperature is -25℃ to 120℃, and the reaction time is 0.5 to 20 h;

[0024] ix) Intermediate 10 undergoes silylation and acetylation reactions to generate compound 11; the base is selected from: sodium hydroxide, potassium carbonate, potassium hydroxide, triethylamine, ethylenediamine, pyridine; the reaction temperature is -20℃ to 80℃, and the reaction time is 0.5 to 20 h;

[0025] x) Intermediate 11 reacts under oxidizing conditions to generate compound 12. The oxidizing agent is selected from: pyridine chlorochromate, pyridine dichromate, Desmartin oxidant, 2-iodobenzoic acid, and manganese dioxide. The reaction temperature is -20℃ to 30℃, and the reaction time is 0.5 to 40 h.

[0026] xi) Deprotection oxidation reaction of intermediate 12, the oxidant is selected from: pyridine chlorochromate, pyridine dichromate, Desmartin oxidant, 2-iodobenzoic acid, manganese dioxide, hydrogen peroxide, peracetic acid, m-chloroperoxybenzoic acid; the reaction temperature is -20℃-30℃, and the reaction time is 0.5-40h;

[0027] xii) Intermediate 11 undergoes an intramolecular Aldol reaction to simultaneously yield compounds with structures of formula Ia, Ib, Ic, and Id; the organic base in the Aldol reaction is selected from: triethylamine, ethylenediamine, N,N-dimethylethylenediamine, and propylenediamine; the reaction solvent is selected from: dichloromethane, tetrahydrofuran, methanol, and N,N-dimethylformamide; the reaction temperature is -65℃ to 50℃, and the reaction time is 0.5 to 20 h.

[0028] Compounds of formulas Ia, Ib, Ic, and Id, or their isomers and pharmaceutically acceptable salts, include: hydrochlorides, hydrobromic acids, phosphates, sulfates, methanesulfonates, p-toluenesulfonates, acetates, trifluoroacetates, salicylates, amino acid salts, citrates, maleates, tartrates, fumarates, citrates, lactates, sodium salts, potassium salts, calcium salts, magnesium salts, lithium salts, ammonium salts, and salts of organic bases that provide physiologically acceptable cations, such as salts of methylamine, dimethylamine, trimethylamine, piperidine, morpholine, and tris(2-hydroxyethyl)amine. All salts of this invention can be prepared using conventional methods. Furthermore, polycrystalline or eutectic crystals may occur under different crystallization conditions during the preparation of solvates of compounds of formulas Ia, Ib, Ic, and Id and their salts.

[0029] Another aspect of the invention relates to a pharmaceutical composition comprising an effective dose of the compound of the invention and a pharmacodynamically acceptable carrier. The pharmaceutical composition can be prepared according to methods known in the art. It can be formulated into any dosage form suitable for human or animal use by combining the compound of the invention with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants. The content of the compound of the invention in its pharmaceutical composition is typically 0.1-95% by weight.

[0030] The compounds of the present invention or pharmaceutical compositions containing them can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0031] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0032] The compounds of this invention can be formulated into conventional formulations, as well as sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0033] To formulate the compounds of the present invention into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0034] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0035] To formulate the drug delivery unit into capsules, the active ingredient, the compound of the present invention, can be mixed with a diluent and a disintegrant, and the mixture can be placed directly into hard or soft capsules. Alternatively, the active ingredient, the compound of the present invention, can be first formed into granules or microspheres with a diluent, binder, and disintegrant, and then placed into hard or soft capsules. The diluents, binders, wetting agents, disintegrants, and disintegrants used to prepare tablets of the compound of the present invention can also be used to prepare capsules of the compound of the present invention.

[0036] To prepare the compounds of this invention into injectable formulations, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure regulators can be added. Solubilizers or co-solvents can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; pH adjusters can be phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; osmotic pressure regulators can be sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing lyophilized powder injections, mannitol, glucose, etc., can also be added as supporting agents.

[0037] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.

[0038] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0039] According to the present invention, compounds of formulas Ia, Ib, Ic, and Id, or their isomers and pharmaceutically acceptable salts thereof, exhibit excellent effects in the treatment of renal fibrosis. Therefore, they can be used as anti-renal fibrosis drugs in animals, preferably in mammals, particularly humans. The compounds of the present invention can be used in the preparation of drugs for the prevention and / or treatment of renal failure caused by renal fibrosis.

[0040] The dosage of the pharmaceutical compositions of the present invention can vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route of administration, and the dosage form. Generally, the suitable daily dose range of the compounds of the present invention is 0.001-150 mg / kg body weight, preferably 0.1-100 mg / kg body weight, more preferably 1-70 mg / kg body weight, and most preferably 2-30 mg / kg body weight. The above dosage can be administered as a single dose unit or divided into several dose units, depending on the physician's clinical experience and the administration regimen, including the use of other treatment methods.

[0041] The compounds or compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs. When the compounds of the present invention have a synergistic effect with other therapeutic drugs, their dosage should be adjusted according to the actual situation.

[0042] Beneficial technical effects

[0043] The compounds of this invention are 9-demethylated natural products of the anti-renal fibrosis compound Commiphorane CD and its isomers. Changes in the configuration of the four chiral centers in the isomers significantly affect their activity. These compounds exhibit good binding to multiple target proteins, good bioavailability, and low toxicity, making them suitable for chronic renal failure and immune-related diseases caused by renal fibrosis. This method provides a way to prepare the C9-demethylated natural product of the anti-renal fibrosis compound Commiphorane CD and its isomers. The method is characterized by constructing the chiral centers at positions 5 and 6 using an asymmetric Aldol reaction, followed by the construction of benzofuran via a HECK reaction. Finally, the route utilizes an intramolecular Aldol reaction to construct the chiral centers at positions 2 and 3, ultimately yielding the 9-demethylated natural product Commiphorane CD and its isomers. This method is original, simple to operate, and convenient for large-scale preparation. Attached Figure Description

[0044] Figure 1 Results of Western blotting analysis of compounds Ia, Ib, Ic, and Id for Fibronectin, Collagen1, α-SMA, and α-Tublin proteins.

[0045] Note: cpd1 is Ia, cpd2 is Ib, cpd3 is Ic, cpd4 is Id, and cpd5 is the control. Detailed Implementation

[0047] The following examples and drug activity experiments are used to further illustrate the present invention, but they do not imply any limitation on the present invention.

[0048] The following examples are used to explain the present invention, but are not intended to limit the invention in any way.

[0049] The raw materials used are known compounds or prepared by known methods.

[0050] Step i):

[0051] Compound 1 (5.8 g, 20 mmol) was dissolved in 100 mL of dichloromethane in a 250 mL three-necked flask. (2.31 mL, 21 mmol) TiCl4 was added at 0°C, and after 15 min, (3.83 mL, 22 mmol) DIPEA was added. The mixture was stirred for 40 min, followed by (1.93 mL, 20 mmol) NMP. The mixture was stirred for 10 min, and finally (5.8 g, 20 mmol) Compound 2 was added at -78°C. The mixture was incubated overnight at room temperature. Saturated ammonium chloride was added at -78°C, and the mixture was extracted with ethyl acetate, washed with saturated sodium bicarbonate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (PE:EA = 20:1) to obtain 8.1 g of Compound 3.

[0052]

[0053] 1 H NMR(400MHz, CDCl3)δ7.48(d,J=7.0Hz,2H),7.40–7.27(m,8H),7.22(d,J=5.0Hz,2H),6.91(d d,J=6.3,3.3Hz,1H),5.46(t,J=2.4Hz,1H),5.16(s,2H),5.04(qd,J=7.0,2.7Hz,1H),4.97(td ,J=7.4,6.8,3.2Hz,1H),4.31(dd,J=9.3,2.4Hz,1H),4.25(d,J=16.1Hz,1H),3.84(dd,J=2.2, 1.1Hz, 1H), 3.23 (dd, J=13.4, 3.6Hz, 1H), 2.79 (dd, J=13.4, 9.9Hz, 1H), 1.27 (d, J=7.0Hz, 3H).

[0054] Step ii):

[0055] Compound 3 (6.67 g, 12.4 mmol) was dissolved in 50 mL of dichloromethane. 2,6-Lutidine (3.9 mL, 33.5 mmol) was added dropwise at -25 °C, followed by the addition of TESOTf. The reaction was maintained at this temperature for 2 h, then stopped by saturated sodium bicarbonate. The mixture was extracted with ethyl acetate, washed with 1 N HCl, washed with saturated sodium chloride, dried over anhydrous magnesium sulfate, filtered, concentrated, and subjected to column chromatography (PE:EA = 30:1) to give compound 4, 6.9 g, in 85.4% yield.

[0056]

[0057] 1H NMR (400MHz, CDCl3) δ7.35–7.25(m,5H),7.18–7.03(m,7H),6.75(dd,J=7.9,1.7Hz,1H),5.30 (d,J=8.4Hz,1H),5.17(d,J=12.6Hz,1H),5.09–4.97(m,2H),4.13(dddd,J=10.5,7.1,3.4,1. 6Hz,1H),3.78(dd,J=9.1,1.7Hz,1H),3.19(dd,J=13.3,3.4Hz,1H),3.04(t,J=8.1Hz,1H),2. 55(dd,J=13.3,10.5Hz,1H),1.43(d,J=6.6Hz,3H),0.82(t,J=7.9Hz,9H),0.58–0.41(m,6H).

[0058] Step iii):

[0059] Compound 4 (6.5 g, 10.0 mmol) was added to 100 mL of dichloromethane at -78 °C with 20 mL of DIBAL-H. After reacting for 1 h, saturated sodium potassium tartrate was added, stirred, separated, and the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude compound 5, 4.6 g of oil, which was used directly in the next step.

[0060]

[0061] 1 H NMR (500MHz, CDCl3) δ9.86 (s, 1H), 7.52 (t, J = 8.3Hz, 3H), 7.46–7.33 (m, 5H), 7.21 (dd, J = 7. 1,4.6Hz,4H),6.92(d,J=8.3Hz,1H),5.79(d,J=2.9Hz,1H),5.33(s,1H),5.24–5.14(m,2H) ,4.77(t,J=8.8Hz,1H),4.46(dd,J=9.0,6.3Hz,1H),4.31(t,J=7.4Hz,1H),3.07–2.87(m,2 H),2.84–2.71(m,1H),1.04(d,J=7.0Hz,3H),0.90(t,J=7.9Hz,9H),0.56(q,J=8.0Hz,6H).

[0062] Step iv):

[0063] Potassium tert-butoxide (2.4 g, 21.4 mmol) was added to 100 mL of tetrahydrofuran at 0 °C. After 20 min, a 10 mL tetrahydrofuran solution of compound 5 (4.5 g, 9.7 mmol) was added, and the reaction was carried out at 0 °C for 30 min. Saturated ammonium chloride was added, and the mixture was diluted with 300 mL of ethyl acetate. Water was added, and the mixture was washed with 50 mL of saturated sodium chloride. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (PE:EA = 50:1) to obtain 4 g of the product, with a yield of 89.2%.

[0064]

[0065] 1 H NMR (400MHz, CDCl3) δ7.53–7.46(m,2H),7.42–7.35(m,2H),7.34–

[0066] 7.29(m,1H),7.21–7.10(m,2H),6.82(dd,J=7.9,1.7Hz,1H),5.86(ddd,J=17.6,10.3,7.6Hz,1H),5.14(s,2H),5.08(d,J=4. 6Hz,1H),5.03–4.89(m,2H),2.51(q,J=6.4Hz,1H),0.96(d,J=6.8Hz,3H),0.85(t,J=7.9Hz,9H),0.50(qd,J=7.9,2.2Hz,6H).

[0067]

[0068] 1 H NMR (400MHz, CDCl3) δ7.53–7.45(m,2H),7.42–7.35(m,2H),7.34–

[0069] 7.28(m,1H),7.21–7.11(m,2H),6.83(dd,J=8.0,1.6Hz,1H),5.14(d,J=2.6Hz,2H),5.07(d,J=3.9Hz,1H),3.76(dq,J=7.2,5.3Hz,1H),3.70 –3.60(m,1H),1.91(m,1H),1.77–1.65(m,1H),1.59–1.48(m,1H),1.36(t,J=5.6Hz,1H),0.86(t,J=7.9Hz,9H),0.51(td,J=8.0,7.0Hz,6H).

[0070] Step vi):

[0071] Compound 7 (2.1 g, 4.4 mmol) was dissolved in 50 mL of ethyl acetate, and 200 mg of Pd-C was added. The mixture was incubated at room temperature for 10 h. TLC showed complete reaction. The mixture was filtered, concentrated, and compound 8 (900 mg) was obtained, with a yield of 75%.

[0072]

[0073] 1 H NMR (400MHz, CDCl3) δ7.07(t,J=7.8Hz,1H),6.95(dd,J=7.8,1.6Hz,1H),6.75(dd,J=8.0,1.6Hz,1H),4.93(d, J=3.6Hz,1H),3.71–3.50(m,2H),2.00(ddt,J=10.3,6.7,3.6Hz,1H),1.74–1.51(m,2H),0.75(d,J=7.0Hz,3H).

[0074] Step vii):

[0075] Compound 8 (0.7 g, 2.55 mmol) was added to 30 mL of acetone, along with (703 mg, 5.1 mmol) K₂CO₃ and (0.26 mL, 3.1 mmol) allyl bromide. The mixture was refluxed for 4 h, filtered, concentrated, and subjected to column chromatography (PE:EA = 1:1) to give compound 9, 0.75 g solid, yield 93.7%.

[0076]

[0077] 1 H NMR (400MHz, CDCl3) δ7.22-7.26(m,1H),7.15(dd,J=7.7,1.5Hz,1H),6.79(dd,J=8.1,1.5Hz,1 H),6.06(ddt,J=17.2,10.2,5.0Hz,1H),5.49(dq,J=17.3,1.7Hz,1H),5.37–5.24(m,1H),5.13 (t,J=3.1Hz,1H),4.59(dt,J=5.0,1.7Hz,2H),3.77(ddd,J=23.4,11.0,5.7Hz,2H),2.72(d,J= 3.4Hz,1H),2.15(qd,J=6.7,3.4Hz,1H),1.87(s,1H),1.79–1.68(m,2H),0.83(d,J=6.9Hz,3H).

[0078] Step viii):

[0079] Compound 9 (0.8 g, 2.4 mmol) was added sequentially to 60 mL of dioxane with cesium fluoride (0.74 g, 4.87 mmol) and Pd(dba)3 (112 mg, 0.12 mmol). The reaction was carried out at 80 °C–100 °C for 16 h. The mixture was diluted with 300 mL of ethyl acetate, washed with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (PE:EA = 2:1) to give 0.4 g of the product, with a yield of 66.7%.

[0080]

[0081] 1 H NMR (400MHz, CDCl3) δ7.35 (dd, J=8.1, 1.0Hz, 2H), 7.23-7.27 (m,

[0082] 1H),5.30(d,J=4.3Hz,1H),3.77(ddd,J=10.7,6.8,5.6Hz,1H),3.66(ddd,J=10.8,6.7,5.6Hz,1H),2.34(d,J =1.4Hz,3H),2.08(tdd,J=7.0,5.4,4.3Hz,1H),1.71(dtd,J=9.2,7.0,6.3,4.6Hz,2H),0.92(d,J=6.8Hz,3H). 13 C NMR (101MHz, CDCl3) δ157.2,143.0,138.3,125.0,124.0,119.7,115.2,110.0,70.1,58.2,37.6,37.2,13.2,11.3.

[0083] Step ix):

[0084] Compound 10 (200 mg, 0.85 mmol) was reacted in 10 mL of dichloromethane with imidazole (116 mg, 1.71 mmol) and TBSCl (141 mg, 0.94 mmol) added in portions. The reaction was carried out at 0 °C to room temperature for 1 h. 100 mL of EA and 20 mL of water were added, followed by washing with 1 N HCl, saturated sodium chloride, and drying over anhydrous sodium sulfate. The mixture was filtered, concentrated, and subjected to column chromatography (PE:EA = 30:1) to give 230 mg of intermediate. This intermediate was dissolved in 10 mL of dichloromethane, and pyridine (0.12 mL, 1.5 mmol) and acetic anhydride (0.14 mL, 1.5 mmol) were added at room temperature. The catalytic amount of DMAP was added, and the reaction was carried out at room temperature for 2 h. 100 mL of EA and 20 mL of water were added, followed by washing with 1 N HCl, saturated sodium chloride, and drying over anhydrous sodium sulfate. The mixture was filtered, concentrated, and subjected to column chromatography (PE:EA = 50:1) to give 190 mg of intermediate, a pale yellow oil with a yield of 57.4%.

[0085]

[0086] 1 H NMR (400MHz, CDCl3) δ7.39–7.31(m,2H),7.27–7.14(m,2H),6.24(d,J=6.6Hz,1H),3.67–3.45(m,2H),2.45(d,J=1.3Hz,3H),2.26 –2.13(m,1H),2.08(s,3H),1.58–1.49(m,1H),1.41–1.23(m,1H),0.97(d,J=6.7Hz,3H),0.82(s,9H),-0.04(s,3H),-0.05(s,3H).

[0087] Step x):

[0088] Compound 11 (160 mg, 0.41 mmol) was dissolved in 5 mL of dichloromethane. 2.5 mL of peracetic acid was added at 0 °C, and the mixture was incubated at room temperature for 4 h. Complete reaction was observed by TLC. 10 mL of saturated sodium thiosulfate was added dropwise at 0 °C, and the mixture was stirred for 10 min. Extraction was then performed with 100 mL of EA, washed with 0.5 N NaOH, washed with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (PE:EA = 30:1) to give compound 12, 0.1 g, yield 60.2%.

[0089]

[0090] 1 H NMR (400MHz, CDCl3) δ7.31–7.21(m,2H),7.10(ddd,J=14.0,8.0,1.0Hz,2H),7.01(ddd,J=7.5,6.1,1.0Hz,2H),5.83(d,J =5.9Hz,1H),5.48(d,J=8.6Hz,1H),4.01(q,J=7.6Hz,1H),3.75–3.52(m,5H),2.35–2.22(m,1H),2.11(s,3H),2.10–2.04 (m, 1H), 2.00 (s, 3H), 1.71 (d, J = 7.5 Hz, 3H), 1.64 (d, J = 7.6 Hz, 3H), 1.53–1.45 (m, 1H), 1.41–1.36 (m, 1H), 1.34–1.27 (m, 1H), 1.23–1.12 (m, 1H), 1.06 (d, J = 6.5 Hz, 3H), 0.90 (d, J = 6.7 Hz, 3H), 0.84 (d, J = 2.9 Hz, 18H), 0.06–-0.08 (m, 11H). (Differential epimer)

[0091] Step xi):

[0092] Compound 12 (57 mg, 0.14 mmol) was added to 5 mL of dichloromethane at 0 °C with (31 μL, 0.28 mmol) 70% hydrofluoric acid pyridine. Complete TLC was observed at this temperature for 0.5 h. 10 mL of saturated sodium bicarbonate was added dropwise, followed by extraction with 50 mL of EA, washing with 10 mL of saturated sodium chloride, drying to anhydrous sodium sulfate, filtration, concentration, and column chromatography (PE:EA = 5:1) to give intermediate 25 mg. Intermediate (25 mg, 0.086 mmol) was added to 5 mL of dichloromethane with 44 mg of Diesel oxidant. Complete TLC was observed at room temperature for 1 h. 3 mL of saturated sodium thiosulfate and 3 mL of saturated sodium bicarbonate were added at 0 °C, followed by extraction with 50 mL of EA, washing with 1 N HCl, washing with 10 mL of saturated sodium chloride, drying to anhydrous sodium sulfate, filtration, concentration, and column chromatography (PE:EA = 5:1) to give compound 13, 20 mg, yield 50%.

[0093]

[0094] 1 H NMR (600MHz, CDCl3) δ9.69(t,J=1.1Hz,1H),9.62(d,J=1.4Hz,0H),7.29(t,J=7.9Hz,2H),7.18–6.96(m,3H),5.89(d,J=4.8 Hz,1H),5.51(d,J=8.7Hz,1H),4.01(q,J=7.7Hz,1H),3.84(q,J=7.5Hz,1H),2.83–2.72(m,1H),2.55(dd,J=7.0,1.1Hz,1H), 2.53–2.47 (m, 2H), 2.33 (dd, J = 4.6, 1.4 Hz, 1H), 2.30–2.25 (m, 2H), 2.13 (s, 3H), 2.02 (s, 2H), 1.73 (d, J = 7.6 Hz, 4H), 1.69 (d, J = 7.6 Hz, 2H), 1.56 (s, 2H), 1.30–1.26 (m, 1H), 1.24 (d, J = 3.8 Hz, 1H), 1.13 (d, J = 6.6 Hz, 2H), 0.95 (d, J = 6.7 Hz, 3H). (Differential epimer)

[0095] Step xii):

[0096] 20 mg (0.07 mmol) of compound 13 was dissolved in dichloromethane or acetone in a 20 mL round-bottom flask. Triethylamine (0.01 mL, 0.08 mmol) was added and the mixture was reacted at room temperature for 4 h. The reaction was complete by TLC. After concentration, the product was purified by column chromatography (PE:EA = 5:1) to give 4 mg of compound Ia.

[0097] Example 1:

[0098]

[0099] 1 H NMR (600MHz, CDCl3) δ7.28(t,J=8.0Hz,1H),7.05(d,J=8.0Hz,1H),7.01(d,J=8.1Hz,1H),5.84(d,J=10.5Hz,1H),3.89 –3.75(m,1H),3.53(s,1H),2.26(s,3H),2.15–2.04(m,2H),1.88(ddd,J=10.6,5.0,2.5Hz,1H),1.06(d,J=6.7Hz,3H).

[0100] Example 2:

[0101] 20 mg (0.07 mmol) of compound 13 was dissolved in dichloromethane or acetone in a 20 mL round-bottom flask. Triethylamine (0.01 mL, 0.08 mmol) was added and the mixture was reacted at room temperature for 4 h. The reaction was complete by TLC. After concentration, the product was purified by column chromatography (PE:EA = 5:1) to give 4 mg of compound Ib.

[0102]

[0103] 1 H NMR (600MHz, CDCl3) δ7.30–7.27(m,1H),7.13(d,J=7.9Hz,2H),5.62(d,J=5.0Hz,1H),4.07(dd,J=11.8,3.4Hz,1H),3.53(s,1H ),2.58(ddd,J=14.7,11.6,3.3Hz,1H),2.52–2.41(m,1H),2.08(s,3H),1.89–1.79(m,1H),1.73(s,3H),0.76(d,J=7.4Hz,3H).

[0104] Example 3:

[0105] 20 mL of compound 13 (20 mg, 0.07 mmol) was dissolved in dichloromethane or acetone. Triethylamine (0.01 mL, 0.08 mmol) was added and the mixture was reacted at room temperature for 4 h. The reaction was complete by TLC. After concentration, the product was obtained by column chromatography (PE:EA = 5:1) to give 4 mg of compound Ic.

[0106]

[0107] 1 H NMR (600MHz, CDCl3) δ7.24-7.27(m,1H),7.13(d,J=7.7Hz,1H),7.09(d,J=8.1Hz,1H),5.69(d,J=4.8Hz,1H),4.37(s,1H),2.62(ddd,J= 15.6,4.2,2.3Hz,1H),2.40(ddd,J=8.6,7.6,4.0Hz,1H),2.07(s,3H),1.93(dt,J=15.6,4.1Hz,1H),1.68(s,3H),0.95(d,J=7.6Hz,3H).

[0108] Example 4:

[0109] 20 mL of compound 13 (20 mg, 0.07 mmol) was dissolved in dichloromethane or acetone. Triethylamine (0.01 mL, 0.08 mmol) was added and the mixture was reacted at room temperature for 4 h. The reaction was complete by TLC. After concentration, the product was obtained by column chromatography (PE:EA = 5:1) to give 4 mg of compound Id.

[0110]

[0111] 1 H NMR(600MHz, CDCl3)δ7.28(t,J=8.0Hz,1H),7.07–6.98(m,2H),5.93(d,J=9.9Hz,1H),4.26(s ,1H),2.27(s,3H),2.23–2.16(m,2H),2.15–2.08(m,1H),1.55(m,3H),1.03(d,J=6.3Hz,3H).

[0112] Pharmacological experiments

[0113] Example 1: Western blotting detection of the effects of 9-demethylated anti-renal fibrosis natural product Commiphorane C-D and its isomer on TGF-β1-induced renal fibrosis cells.

[0114] The effect of the 9-demethylated Commiphorane C isomer on TGF-β1-induced renal fibrosis cells was detected by Western blotting. 800,000 NRK52E cells per well were seeded into 6-well plates, induced with TGF-β1, and treated with the compound for 48 h. Proteins were extracted, quantified by BCA, and then subjected to electrophoresis, membrane transfer, antibody incubation, and ECL imaging to determine the expression levels of Fibronectin, Collagen I, and α-SMA proteins in the cells.

[0115] Experimental methods

[0116] Cell Count

[0117] (1) Preheat the culture medium used in cell passage in a 37°C water bath.

[0118] (2) NRK52E cells were digested with trypsin, and culture medium was added to stop the digestion. After the digestion was stopped, the cell suspension was transferred to a 15ml centrifuge tube.

[0119] (3) Count cells using a cell counting chamber. Take 1 million cells and put them into a 1.5ml centrifuge tube.

[0120] Cell protein extraction

[0121] (1) Place a 1.5ml centrifuge tube containing cell suspension in a centrifuge and centrifuge at 4℃, 12000rpm for 15min. Discard the supernatant.

[0122] (2) Take 1 ml of 1x PBS solution to resuspend the cells, centrifuge at 12000 rpm for 15 min at 4℃, and discard the supernatant.

[0123] (3) Repeat step (2).

[0124] (4) Prepare cell lysis buffer and add an appropriate amount of protease inhibitor; add 100 μl of cell lysis buffer to each 1.5 ml centrifuge tube.

[0125] (5) On ice, lyse for 15 min.

[0126] (6) Use a cell ultrasonic disruptor to disrupt the cells at 20% power, sonicate for 2 seconds, pause for 5 seconds, and the whole process lasts for 1 minute.

[0127] (7) Centrifuge at 4℃ and 12000rpm for 15min. After centrifugation, take the supernatant and aliquot it into 1.5mL centrifuge tubes. A portion of the supernatant is used for BCA assay. Take another 80μl of cell lysis buffer, add 20μl of bromophenol blue solution, and place in a metal bath at 100℃ for 10 minutes.

[0128] BCA protein assay

[0129] (1) Preparation of standards: Take 30 μl of 2 μg / μl BSA solution and serially dilute it with 30 μl of 1xPBS to obtain 6 concentration points. The first concentration point is 2 μg / μl BSA solution, and the eighth concentration point is 0 μg / μl BSA solution. Sample preparation: Take 3 μl of sample lysis buffer and add it to 27 μl of 1xPBS to dilute it 10 times.

[0130] (2) Prepare a certain volume of BCA working solution according to the number of samples. BCA working solution preparation: add 1 volume of BCA reagent B to 50 volumes of BCA reagent A (A:B = 50:1) and mix thoroughly.

[0131] (3) Take 20 μl of standard and test sample into a 96-well plate.

[0132] (4) Add 200 μl of BCA working solution to each well, shake for 30 seconds to mix thoroughly, cover the microplate, and incubate at 37°C for 30 minutes.

[0133] (5) After incubation, the absorbance was measured at 570 nm using an ELISA reader.

[0134] Electrophoresis, membrane transfer

[0135] (1) After washing and drying the glass plate, fix it on the gel casting machine. Pour the separating gel into the gaps of the glass plate to an appropriate height, add an appropriate amount of anhydrous ethanol, and remove air bubbles from the gel surface. After the separating gel has completely solidified, pour out the anhydrous ethanol, then add the stacking gel to an appropriate height, insert the comb teeth, and remove the comb teeth after the stacking gel has completely solidified.

[0136] (2) Electrophoretic separation: Fix the prepared gel onto the electrophoresis tank and pour the electrophoresis buffer into the reservoir. Use a micropipette to add the prepared protein sample and marker to the sample wells, with a sample volume of 15 μl. After adding the sample, electrophoresis is performed at a constant voltage of 60 V until the bromophenol blue indicator forms a line at the junction of the stacking gel and the separating gel (about 30 min). Then, the voltage is changed to a constant voltage of 120 V until the bromophenol blue reaches the bottom of the gel (about 1 h). This process takes about 1.5 h.

[0137] (3) Transfer: Remove the gel and discard the stacking gel. Rinse off any residual electrophoresis buffer on the separating gel with distilled water. Cut a PVDF membrane (soaked in methanol for 1-3 minutes) and filter paper to the same size as the PAGE gel and soak them together in the electrotransfer buffer. Arrange the gel in the following order: black plate - fiber pad - filter paper - gel - PVDF membrane - filter paper - fiber pad - white plate (gel negative, membrane positive). Clamp the plates and place them in the transfer apparatus, with the black plate facing the black negative electrode. Fill the transfer tank with the electrotransfer buffer and begin the transfer. It is best to place the electrophoresis tank on crushed ice during the transfer to avoid excessive temperature. Constant current: 300mA, 60 minutes.

[0138] Closed

[0139] (1) Remove the PVDF membrane and wash it: TBST 3 times, 5 min each time.

[0140] (2) Soak the PVDF membrane in TBST (blocking solution) containing 5% skim milk powder, block on a shaker at 37°C for 1.0 h, and incubate with primary antibody overnight.

[0141] Dilute the corresponding primary antibody with TBST or 5% skim milk powder, immerse the PVDF membrane in the primary antibody incubation solution, and incubate overnight at 4°C (in an ice box).

[0142] Secondary antibody incubation

[0143] (1) Washing the membrane: TBST 3 times, 5 minutes each time;

[0144] (2) Dilute the corresponding secondary antibody with TBST, immerse the PVDF membrane in the secondary antibody incubation solution, and incubate at room temperature for 2 hours;

[0145] (3) Washing membrane: TBST 3 times, 5 minutes each time

[0146] Color Developing Exposure

[0147] Mix the enhancement solution and stable peroxidase solution in the ECL reagent at a 1:1 ratio, add an appropriate amount of working solution to the PVDF membrane, and develop.

[0148] Experimental results

[0149] Bio-Rad gel imaging revealed bands of Fibronectin, Collagen1, α-SMA, and α-Tublin in NRK52E cells. The grayscale values ​​of each protein and α-Tublin were calculated based on the bands. Results showed that after co-incubation of NRK52E cells with TGF-β1 (10 ng / ml) and GW788388 (25 μM), the expression levels of Fibronectin and α-SMA proteins were significantly reduced, but the reduction in Collagen1 expression was not significant. After incubation of NRK52E cells with TGF-β1 (10 ng / ml) and different compounds for 48 hours, Id and cmp5 significantly downregulated Fibronectin expression, while other compounds had little effect on Fibronectin expression. Id downregulated α-SMA expression and also had a certain downregulating effect on Collagen1 expression. (Repeat experiments showed that Ib, Ic, and Id reduced the expression level of α-SMA protein.) Specific data are shown in Table 1.

[0150] Table 1. Gray values ​​of Fibronectin, Collagen1, α-SMA, and α-Tublin

[0151]

Claims

1. Compounds, isomers, or pharmaceutically acceptable salts thereof as indicated by formulas Ia, Ib, Ic, and Id.

2. The method for preparing the compound according to claim 1, characterized in that, The steps include: (1) Methods for preparing formulas Ia, Ib, Ic, and Id: step: i) Compound 1 and compound 2 generate intermediate 3 via asymmetric Aldol; ii) Intermediate 3 is reacted with trimethylchlorosilane to give intermediate 4; iii) Intermediate 4 is hydrolyzed under organic or inorganic alkaline conditions to generate compound 5; iv) Compound 5 reacts with methyltriphenylphosphine bromide under alkaline conditions to give compound 6; v) Compound 6 undergoes a hydroboration oxidation reaction to generate intermediate 7; vi) Intermediate 7 is reduced on palladium on carbon to obtain intermediate 8; vii) Intermediate 8 undergoes a substitution reaction with allyl bromide to generate intermediate 9; viii) Intermediate 9 is converted into intermediate 10 under alkaline conditions and with the participation of a Pd catalyst; (ix) Intermediate 10 undergoes a substitution reaction to generate compound 11; x) Compound 11 undergoes an oxidation reaction to generate intermediate 12; xi) Intermediate 12 undergoes deprotection oxidation to generate intermediate 13; xii) Intermediate 13 can be reacted intramolecularly with Aldol to simultaneously yield compounds with structures of formulas Ia, Ib, Ic and Id.

3. The preparation method according to claim 2, characterized in that, (1) The asymmetric Aldol reaction in step i) occurs in the presence of dichloromethane, tetrahydrofuran, toluene, methanol, and dioxane solvent; (2) The solvent in step ii) is selected from: dichloromethane, tetrahydrofuran, and toluene; (3) The organic base in step iii) is selected from: triethylamine, ethylenediamine, N,N-dimethylethylenediamine, propylenediamine, and diisopropyl lithium hydride; the inorganic base is selected from: potassium carbonate, sodium carbonate, cesium carbonate, cesium fluoride, and lithium hydroxide. (4) The alkali in step iv) is selected from: potassium carbonate, sodium carbonate, cesium carbonate, cesium fluoride, triethylamine, ethylenediamine, N,N-dimethylethylenediamine, propylenediamine, potassium tert-butoxide; (5) The boron reagent used in step v) of the borohydride oxidation reaction is selected from: boron tetrahydrofuran complex, boron dimethyl sulfide complex, and 9-BBN (9-boron bicyclo(3,3,1)-nonane); (6) The reducing reagent used in step vi) is selected from: palladium on carbon and palladium hydroxide on carbon; (7) In step vii), the alkylation reaction, the base is selected from: sodium hydroxide, potassium carbonate, potassium hydroxide, triethylamine, ethylenediamine, pyridine; (8) In step viii) cyclization reaction, the Pd catalyst is selected from: bis(dibenzylacetone palladium), palladium acetate, and 1,1′-bis(diphenylphosphine)ferrocene palladium dichloride; (9) In step ix), the silylation and acetylation reactions, the base is selected from: sodium hydroxide, potassium carbonate, potassium hydroxide, triethylamine, ethylenediamine, and pyridine; (10) The oxidation reaction in step x) is carried out by an oxidant selected from: pyridine chlorochromate, pyridine dichromate, Desmartin oxidant, 2-iodobenzoic acid, and manganese dioxide. (11) The deprotecting oxidation reaction in step xi) uses an oxidant selected from: pyridine chlorochromate, pyridine dichromate, Desmartin oxidant, 2-iodobenzoic acid, manganese dioxide, hydrogen peroxide, peracetic acid, and m-chloroperoxybenzoic acid. (12) The Aldol reaction in step xii) uses an organic base selected from: triethylamine, ethylenediamine, N,N-dimethylethylenediamine, and propylenediamine.

4. The preparation method according to claim 2, characterized in that: (1) The asymmetric Aldol reaction in step i) occurs in the presence of dichloromethane, tetrahydrofuran, toluene, methanol and dioxane solvent, at a reaction temperature of -80℃ to 25℃ and a reaction time of 0.5-10h. (2) The reducing agent in step ii) is selected from: the reaction solvent is selected from: tetrahydrofuran, dichloromethane; the reaction temperature is: -78℃-25℃, and the reaction time is 0.5-10h; (3) In step iii), the organic base is selected from: triethylamine, ethylenediamine, N,N-dimethylethylenediamine, propylenediamine, and diisopropyl lithium hydride; the inorganic base is selected from: potassium carbonate, sodium carbonate, cesium carbonate, cesium fluoride, and lithium hydroxide; the reaction solvent is selected from: acetone, tetrahydrofuran, dichloromethane, and toluene; the reaction temperature is -78℃ to 25℃, and the reaction time is 0.5 to 10 h; (4) In step iv), the base is selected from: potassium carbonate, sodium carbonate, cesium carbonate, cesium fluoride, triethylamine, ethylenediamine, N,N-dimethylethylenediamine, propylenediamine, potassium tert-butoxide; the reaction solvent is selected from: dioxane, N,N-dimethylformamide, acetonitrile, water, ethylene glycol dimethyl ether, tetrahydrofuran, diethyl ether; the reaction temperature is -60℃ to 50℃, and the reaction time is 1 to 30 h; (5) The reagents used in step v) of the borohydride oxidation are selected from: borane tetrahydrofuran complex, borane dimethyl sulfide complex, 9-BBN (9-boron bicyclo(3,3,1)-nonane), and the reaction solvents are selected from: dichloromethane, tetrahydrofuran, methanol, N,N dimethylformamide, and water; the reaction temperature is -78℃ to 50℃, and the reaction time is 0.5 to 10 h; (6) The reduction reaction in step vi) uses a reducing reagent selected from palladium on carbon and palladium hydroxide on carbon; the reaction temperature is -25℃ to 40℃ and the reaction time is 0.5 to 20h. (7) In step vii), the alkylation reaction is carried out using a base selected from sodium hydroxide, potassium carbonate, potassium hydroxide, triethylamine, ethylenediamine, and pyridine. The reaction temperature is -25℃ to 40℃, and the reaction time is 0.5 to 20 h. (8) In step viii) the cyclization reaction, the Pd catalyst is selected from: bis(dibenzylacetone)palladium, palladium acetate, 1,1′-bis(diphenylphosphine)ferrocene palladium dichloride; the reaction solvent is selected from: dichloromethane, tetrahydrofuran, methanol, N,N-dimethylformamide, dioxane, toluene, water; the reaction temperature is -25℃ to 120℃, and the reaction time is 0.5-20h; (9) In the silylation and acetylation reactions of step ix), the base is selected from: sodium hydroxide, potassium carbonate, potassium hydroxide, triethylamine, ethylenediamine, pyridine; the reaction temperature is -20℃ to 80℃, and the reaction time is 0.5 to 20h; (10) The oxidation reaction in step x) is carried out by oxidizing agent selected from: pyridine chlorochromate, pyridine dichromate, Desmartin oxidizing agent, 2-iodobenzoic acid, manganese dioxide; the reaction temperature is -20℃-30℃, and the reaction time is 0.5-40h. (11) The deprotecting oxidation reaction in step xi) uses an oxidant selected from: pyridine chlorochromate, pyridine dichromate, Desmartin oxidant, 2-iodobenzoic acid, manganese dioxide, hydrogen peroxide, peracetic acid, and m-chloroperoxybenzoic acid; the reaction temperature is -20℃ to 30℃ and the reaction time is 0.5 to 40h. (12) The Aldol reaction in step xii) uses an organic base selected from: triethylamine, ethylenediamine, N,N-dimethylethylenediamine, and propylenediamine; the reaction temperature is -65℃ to 50℃ and the reaction time is 0.5 to 20h.

5. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises effective doses of Ia, Ib, Ic and Id as described in claim 1, isomers or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers or excipients.

6. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition is selected from tablets, capsules, pills, and injections.

7. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition is selected from sustained-release formulations, controlled-release formulations, and various microparticle delivery systems.

8. The use of the Ia, Ib, Ic and Id, isomers or pharmaceutically acceptable salts thereof as described in claim 1 in the preparation of medicaments for the treatment and / or prevention of chronic renal failure caused by renal fibrosis.

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