CN117919222A - Application of 3-oxo-benzofuran compounds in preparation of medicines for treating heart failure related diseases - Google Patents
Application of 3-oxo-benzofuran compounds in preparation of medicines for treating heart failure related diseases Download PDFInfo
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- CN117919222A CN117919222A CN202410112890.9A CN202410112890A CN117919222A CN 117919222 A CN117919222 A CN 117919222A CN 202410112890 A CN202410112890 A CN 202410112890A CN 117919222 A CN117919222 A CN 117919222A
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- oxo
- heart failure
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- benzofuran compound
- benzofuran
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- 241000701161 unidentified adenovirus Species 0.000 description 1
- 239000001052 yellow pigment Substances 0.000 description 1
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Abstract
The invention belongs to the technical field of biological medicines, and particularly relates to application of a 3-oxo-benzofuran compound in preparation of a medicine for treating heart failure related diseases. Experiments show that the 3-oxo-benzofuran compound LC-4 can play a remarkable role in myocardial protection in a doxorubicin (Dox) induced heart injury model, an angiotensin II (Ang II) induced heart injury model and an Isoprenaline (ISO) induced chronic heart failure model; the mechanism of the compound LC-4 is that the SNX3 subcellular localization is changed by combining with SNX3, and the combination of the SNX3 and the VPS35 is interfered, so that the SNX3 function is inhibited, the downstream pathway is further inhibited, the heart fibrosis and heart failure caused by the heart fibrosis can be improved, and the effect of preventing and treating the heart failure is finally achieved.
Description
Technical Field
The invention belongs to the technical field of biological medicine. More particularly relates to application of 3-oxo-benzofuran compounds in preparing medicaments for treating heart failure related diseases.
Background
Heart failure is a myocardial injury caused by myocardial infarction, cardiomyopathy, overload of hemodynamic load, inflammation and the like, which can cause changes in myocardial structure and function, and finally, ventricular pump blood or filling function is low. Heart failure is clinically primarily characterized by hemodynamic changes, particularly in pulmonary or systemic congestion and tissue hypoperfusion, which may cause dyspnea, weakness and fluid retention. Chronic heart failure refers to a sustained heart failure state, which is the final stage in patients with various heart related diseases. The aim of treating heart failure is to improve the hemodynamics for a short period, alleviate clinical symptoms, improve the quality of life, and also aim at the mechanism of myocardial reconstruction, prevent or delay the occurrence and development of myocardial reconstruction, improve the biological function of the heart muscle, and reduce the hospitalization rate and the death rate of heart failure.
Currently, the latest heart failure guidelines recommend an angiotensin receptor enkephalinase inhibitor or angiotensin converting enzyme inhibitor/angiotensin ii receptor antagonist, a sodium-glucose cotransporter 2 inhibitor, a beta receptor blocker, and a mineralocorticoid receptor antagonist as a "new quadruple" regimen for heart failure drug treatment to be widely used in clinical treatment, and a great deal of clinical research has been conducted to confirm that the treatment regimen can reduce the mortality and morbidity of chronic heart failure patients (the society of cardiovascular physicians and departments of the Chinese physician association, the center alliance of heart failure in China, the working group of clinical decision path expert consensus for "new quadruple" drug treatment for chronic heart failure, etc. chronic heart failure "new quadruple" drug treatment clinical decision path expert consensus [ J ]. Chinese journal of circulation, 2022,37 (8): 13. However, the solution requires multiple drugs to be combined, which is complex, and the adverse effects of the drugs are easily aggravated, and there is still an urgent need to provide more drugs for treating heart failure, so as to provide more treatment options.
Disclosure of Invention
The invention aims to overcome the defects that the existing combined drug for treating heart failure needs a plurality of medicaments to be used in combination, is complex and aggravates adverse reactions of the medicaments easily, and provides the application of the 3-oxo-benzofuran compounds or derivatives thereof in preparing medicaments for treating heart failure related diseases.
The invention aims to provide a composition for treating heart failure related diseases.
The invention also aims to provide an application of the 3-oxo-benzofuran compounds or derivatives thereof in preparing inhibitors of sortilin 3.
It is still another object of the present invention to provide the use of a 3-oxo-benzofuran compound or derivative thereof in the preparation of a transcriptional activator protein 3 phosphorylation inhibitor.
The above object of the present invention is achieved by the following technical scheme:
sortation-connexin 3 (SNX 3) is an antiport-related cargo-binding protein, and has become a key regulator of protein transport involved in the pathogenesis of a variety of diseases. Structurally, SNX3 binds directly or indirectly to the Vacuolar Protein Sorting (VPS) family VPS26, VPS35 and VPS29, constituting an antiporter complex; SNX3 also targets endosomal membranes through Phox homology (PX) domains. Functionally, SNX3 recruits the reverse endosomes to the Early Endosomes (EE) or the Recirculating Endosomes (RE), followed by the budding of vesicles from EE/RE to the Trans Golgi Network (TGN), plasma Membrane (PM), or nucleus-mediated retrograde transport of cargo proteins. Among them, SNX3 is the simplest structural subtype in the sorted connexin family, and it has been reported in the literature that SNX3 causes heart damage by promoting signal transduction and phosphorylation and dimerization of transcription activator protein 3 (STAT 3) and translocation thereof into the nucleus (Lu J,Xu S,Huo Y,et al.Sorting nexin 3induces heart failure via promoting retromer-dependent nuclear trafficking of STAT3[J].Cell death and differentiation,2021(10):28.).
Chinese patent application CN111285829a discloses a class of pparγ/δ dual agonists, wherein compound I-8 is disclosed, namely: (Z) -2- (4- ((6-ethoxy-3-oxo-benzofuran-2 (3H) -subunit) methyl) -2, 6-dimethyl phenoxy-2-methylpropanoic acid is named as a compound LC-4, and the compound has better PPARgamma/delta dual-agonist activity and can prevent or/and treat diabetes, obesity, hyperlipidemia, atherosclerosis, fibrosis, fatty liver and other metabolic syndrome related diseases.
Experiments show that the compound LC-4 can play a remarkable myocardial protection role in an doxorubicin (Dox) induced heart injury model, an angiotensin II (Ang II) induced heart injury model and an Isoprenaline (ISO) induced chronic heart failure model. At present, no drug for treating heart diseases by taking SNX3 as a target point exists, and the invention discovers that the mechanism of the compound LC-4 is that the subcellular localization of the SNX3 is changed by combining with the SNX3, and the combination of the SNX3 and the VPS35 is interfered, so that the SNX3 function is inhibited, the downstream passage is further inhibited, the heart fibrosis and heart failure caused by the heart fibrosis can be improved, and the effect of preventing and treating the heart failure is finally achieved.
Therefore, the invention claims the application of 3-oxo-benzofuran compounds or derivatives thereof in preparing medicaments for treating heart failure related diseases, wherein the 3-oxo-benzofuran compounds have the following structures:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
Further, the pharmaceutically acceptable salt of the 3-oxo-benzofuran compounds is a salt formed by the 3-oxo-benzofuran compounds and organic bases or inorganic bases.
Preferably, the organic base is triethylamine, pyridine, sodium tert-butoxide, potassium tert-butoxide, diisopropylamine, n-butyllithium, isobutyllithium, tert-butyllithium, sodium methoxide, sodium ethoxide or sodium amide.
Preferably, the inorganic base is sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium thiosulfate, sodium hydroxide, lithium hydroxide, potassium hydroxide, ammonia water or ammonium bicarbonate.
Further, the heart failure-related diseases include coronary heart disease, myocardial infarction, angina pectoris, myocarditis, cardiac fibrosis, and the like.
Further, the medicament also comprises pharmaceutically acceptable auxiliary materials.
Preferably, the auxiliary materials are selected from one or more of diluents, excipients, disintegrants, fillers, binders, lubricants, flavoring agents, suspending agents, surfactants and stabilizers.
Further, the dosage form of the medicine is tablets, capsules, pills, powder, granules, solutions or injections.
In addition, based on heart failure guideline recommended medication, the compound LC-4 of the invention can be combined with known anti-heart failure medicines to achieve the effect of treating heart failure related diseases. Accordingly, the present invention also claims a composition for treating heart failure related diseases, the composition comprising a 3-oxo-benzofuran-type compound or derivative thereof and an anti-heart failure drug; the 3-oxo-benzofuran compounds have the following structures:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
Further, the heart failure resistant drug is selected from one or more of an angiotensin receptor enkephalinase inhibitor or an angiotensin converting enzyme inhibitor/angiotensin II receptor antagonist, a sodium-glucose cotransporter 2 inhibitor, a beta receptor blocker and a mineralocorticoid receptor antagonist.
Based on that the compound LC-4 can interfere with the binding of SNX3 to VPS35 by altering the subcellular localization of SNX3 by binding to SNX3, thereby inhibiting the function of SNX3, the invention also claims the use of a 3-oxo-benzofuran compound or derivative thereof for the preparation of a split-junction protein 3 (SNX 3) inhibitor, said 3-oxo-benzofuran compound having the following structure:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
Preferably, the 3-oxo-benzofuran compounds are inhibitors of sortilin 3 (SNX 3).
Based on the fact that the compound LC-4 can inhibit STAT3 phosphorylation, the invention also claims the application of a 3-oxo-benzofuran compound or a derivative thereof in preparing a transcription activator protein 3 (STAT 3) phosphorylation inhibitor, wherein the 3-oxo-benzofuran compound has the following structure:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
The invention has the following beneficial effects:
Experiments show that the 3-oxo-benzofuran compound LC-4 can play a remarkable role in myocardial protection in a doxorubicin (Dox) induced heart injury model, an angiotensin II (Ang II) induced heart injury model and an Isoprenaline (ISO) induced chronic heart failure model; the mechanism of the compound LC-4 is that the SNX3 subcellular localization is changed by combining with SNX3, and the combination of the SNX3 and the VPS35 is interfered, so that the SNX3 function is inhibited, the downstream pathway is further inhibited, the heart fibrosis and heart failure caused by the heart fibrosis can be improved, and the effect of preventing and treating the heart failure is finally achieved.
Drawings
FIG. 1 is a statistical chart of the results of myocardial protection experiments in a Dox cardiomyopathy model for compound LC-4 of example 1; wherein # # # is P <0.001 compared to the CON group; * P <0.01 compared to the Dox group and P <0.001 compared to the Dox group.
FIG. 2 is a Western blot showing inhibition of STAT3 phosphorylation by compound LC-4 under induction of Ang II in example 2.
FIG. 3 is an immunofluorescence and Western blot of experiments demonstrating the effect of the compounds LC-4 and VPS34 inhibitor IN1 of example 3 on the structure of SNX3 protein.
FIG. 4 is an immunofluorescence co-localization map of the effect of compound LC-4 on SNX3 binding to VPS35 in H9C2 cells of example 4.
FIG. 5 is a Western blot of example 5 demonstrating that compound LC-4 affects SNX3 binding to VPS35 in H9C2 cells by co-immunoprecipitation (co-IP) experiments.
FIG. 6 is an immunofluorescence co-localization map of the binding of the compound LC-4-GFP conjugate to SNX3 in H9C2 cells as verified by immunofluorescence experiments in example 6.
FIG. 7 is a Western blot showing the binding of the compound LC-4-GFP conjugate to SNX3 in H9C2 cells by co-immunoprecipitation (co-IP) experiments, example 7.
FIG. 8 is a Western blot of the validation of the inhibition of AngII-stimulated cell fibrosis by compound LC-4 in SD milk mouse primary myocardial fibroblasts, example 8.
FIG. 9 is an echocardiogram (A), ejection Fraction (EF) map (B), left ventricular short axis foreshortening rate (FS) map (C), end-diastolic left ventricular anterior wall thickness (LAVW; D) map (D), end-systolic left ventricular anterior wall thickness (LAVW; s) map (E), end-diastolic left ventricular posterior wall thickness (LAPW; D) map (F), end-systolic left ventricular posterior wall thickness (LAPW; s) map (G), stroke Volume (SV) map (H), cardiac Output (CO) map (I) for example 9 animal model validation compound LC-4 under ISO-induced chronic heart failure model.
FIG. 10 shows the heart morphology size chart (A), HE staining chart (B), weight to weight ratio (HW/BW) chart (C), weight to length ratio (HW/TL) chart (D), HE staining chart (E), sirius red staining chart (F), wheat Germ Agglutinin (WGA) staining chart (G) of an ISO model laboratory animal in example 10.
FIG. 11 is a Western blot image relating to fibrosis of mice which were model of ISO-induced heart failure in example 10.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
The 3-oxo-benzofuran compound LC-4 reference Li Z,Ren Q,Zhou Z,Cai Z,Wang B,Han J,Zhang L.Discovery of the first-in-class dual PPARδ/γpartial agonist for the treatment of metabolic syndrome.Eur J Med Chem.2021Dec 5;225:113807.doi:10.1016/j.ejmech.2021.113807.Epub 2021Aug 25.PMID:34455359. is prepared.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
Example 13 myocardial protection of oxo-benzofurans LC-4
S1, taking well-grown rat myocardial cell strain H9C2, culturing 5×10 4 cells per well in 96-well transparent flat bottom plates by using high-sugar DMEM,10% fetal bovine serum, 5% carbon dioxide and 37 ℃;
After S2 and 24h adherence, compound LC-4 with different drug concentrations (1.25. Mu.M, 2.5. Mu.M, 5. Mu.M, 10. Mu.M, 20. Mu.M, 40. Mu.M, 80. Mu.M, 160. Mu.M) was given;
S3, after pretreatment for 12 hours, 1 mu M doxorubicin (Dox) is given to stimulate H9C2 cells to establish a Dox myocardial cell injury model;
s4, after culturing for 12 hours, adding a complete culture medium containing 10% of CCK8 solution, incubating for 1-4 hours, measuring absorbance (OD value) at 450nm by using a microplate reader, and calculating cell viability (Cell Vilability).
The experimental result is shown in figure 1, and the experimental result shows that the compound LC-4 can obviously improve the cell activity (Cell Vilability) and restore the cells to the normal state, thereby showing that the compound LC-4 has better myocardial protection effect.
EXAMPLE 23 inhibition of STAT3 phosphorylation of oxo-benzofurans LC-4
S1, taking well-grown rat myocardial cell strain H9C2, culturing 5×10 4 cells per well in 96-well transparent flat bottom plates by using high-sugar DMEM,10% fetal bovine serum, 5% carbon dioxide and 37 ℃;
S2, after the cells grow to 70% -80% of density, starving the cells by using a high-sugar DMEM culture medium containing 1% of fetal calf serum;
S3, after starvation treatment, replacing the high-sugar DMEM containing 10% of fetal bovine serum, and simultaneously pre-dosing compound LC-4 with different concentrations (1 mu M, 2.5 mu M, 5 mu M and 10 mu M);
S4, after pre-administration for 12 hours, 1 mu M angiotensin II (Ang II) is administrated to stimulate H9C2 cells for 12 hours to induce STAT3 phosphorylation;
and S5, collecting cell proteins after 12 hours, and performing Western blot experiment (Western blot) detection after quantitative determination by using BCA.
The experimental results are shown in fig. 2, and it can be seen from the experimental results that the compound LC-4 can significantly inhibit STAT3 phosphorylation caused by Ang II.
Example 33 Effect of an oxo-benzofuran type LC-4 on SNX3 protein localization
S1, taking well-grown rat myocardial cell strains H9C2, adding 5 mu M of compound LC-4 or an inhibitor Vps34-IN-1 (MCE, # 1383716-33-3) of Vps34 into a laser confocal cuvette, and respectively treating for 1-6H;
s2, fixing the mixture for 20min at room temperature by using 4% paraformaldehyde, washing the mixture by using PBS, permeabilizing the mixture for 20min at 37 ℃ by using 1% tween-20, and washing the mixture by using PBS;
S3, blocking for 1h by using goat serum, adding early endo-body antigen 1 antibodies (EEA 1, CST, # 2411) diluted according to a recommended proportion, SNX3 (Santa, # sc-376667) antibodies, and respectively incubating alpha tubulin serving as an internal reference protein (alpha-tubulin, wuhan Sanying, # 66031-1-Ig) overnight, and washing by using PBS;
s4, performing light-shielding operation, adding a fluorescent secondary antibody of a corresponding species for incubation for 1h, washing with PBS, using DAPI dye liquor to dye the nuclei for 10min, and washing with PBS;
s5, observing fluorescence distribution under a laser confocal microscope.
As shown in fig. 3, the permeabilization treatment in the experiment eluted SNX3 not bound to the early endosomal membrane, thereby showing only the remaining SNX3 bound to the early endosomal membrane; the function of SNX3 to play a role in sorting proteins mainly depends on subcellular localization, and from experimental results, it can be seen that the compound LC-4 can dissociate SNX3 from early endosomal membranes into cytoplasm, and change subcellular localization of SNX3 without changing the total protein level and destroying early endosomal structure. The compound LC-4 can regulate subcellular localization of SNX3 to play a role in myocardial protection.
Example 43 Effect of an oxo-benzofuran type LC-4 on the inverse SNX3 transport Complex
S1, taking well-grown rat myocardial cell strain H9C2, putting the well-grown rat myocardial cell strain H9C2 into a laser confocal small dish, and adding 40 mu M of compound LC-4 for treatment for 12 hours;
S2, processing according to the method in the embodiment 3, wherein 1% Tween-20 permeabilization is changed into 0.3% Triton X-100 permeabilization for 10min at room temperature;
s3, adding VPS35 (Wuhan Sanying, #10236-1-AP,1:200 dilution) and SNX3 antibody diluted according to the recommended proportion, respectively incubating overnight, incubating the fluorescent secondary antibody with the operation of example 3, staining with DAPI, and observing fluorescence distribution under a laser confocal microscope.
As shown in FIG. 4, the permeabilization treatment in the experiment did not elute SNX3 from the cytoplasm. From the experimental results, it can be seen that the co-localization coefficient (Pearson , s Rr) and co-localization yellow pigment spot were significantly reduced, indicating that compound LC-4 can inhibit the binding of SNX3 to VPS 35.
Example 53 Effect of an oxo-benzofuran type LC-4 on the inverse SNX3 transport Complex
S1, taking well-grown rat myocardial cell strain H9C2, putting the well-grown rat myocardial cell strain H9C2 into a 10cm cell culture dish, adding adenovirus to overexpress SNX3 (Ad-SNX 3) when the cell density reaches 70% -80%, and dividing the cell density into a CON group and an LC-4 group, wherein the LC-4 group is treated for 12 hours after 40 mu M of compound LC-4 is added, the CON is not treated additionally, and the LC-4 group is treated for 12 hours after 40 mu M of compound LC-4 is added;
S2, extracting total cell proteins for quantification, respectively taking 500 mug of proteins from a CON group and an LC-4 group, adding a VPS35 antibody according to a recommended proportion, simultaneously respectively taking 20 mug of proteins from the total proteins as positive control (Input) samples before co-immunoprecipitation, taking about 100 mug of proteins and adding immunoglobulin G (IgG) of the same species as negative control, and slowly shaking the IgG group and the VPS35 antibody group at 4 ℃ for overnight;
S3, adding 30 μl protein A/G agarose microspheres (Siemens, 88803) into each group of EP tubes, uniformly mixing, slowly shaking at 4 ℃ for 4h, centrifuging at 4 ℃ for 30S at 12000G, and discarding the supernatant;
S4, using an immune coprecipitation Buffer 1 (Buffer l) with gradient of NaCl concentration, using an immune coprecipitation Buffer 2 (Buffer 2), washing and precipitating the immune coprecipitation Buffer 3 (Buffer l) once, centrifuging for 30S at the temperature of 4 ℃ after each washing, discarding the supernatant, and sucking the supernatant as completely as possible each time, but avoiding sucking the supernatant to the precipitation;
S5, adding 30 mu l of SDS-PAGE protein loading buffer into each group of EP pipes, uniformly mixing, and placing in a metal bath at 100 ℃ for boiling for 5min to respectively serve as IP: CON group and LC-4 group of VPS 35;
s6, performing Western blot detection on the Input, IP and IgG control samples.
The experimental results are shown in FIG. 5, and it can be seen from the co-immunoprecipitation (co-IP) experimental results that the SNX3 expression level of the LC-4 group is significantly reduced compared with the CON group, indicating that the compound LC-4 can inhibit the binding of SNX3 to VPS 35.
Example 6 and example 7 were performed by submitting the coupling body of the compound LC-4 and Green Fluorescent Protein (GFP) to the Hebei Langmuir Biotechnology Co., ltd, and the binding between LC-4 and SNX3 was examined by GFP.
Example 6 3 interaction of an oxo-benzofuran-based LC-4-GFP conjugate with SNX3
S1, taking well-grown rat myocardial cell strain H9C2, adding the strain into a laser confocal cuvette, adding Ad-SNX3 virus to overexpress SNX3, adding 1 mu L of conjugate solution after 12 hours to make the final concentration of the conjugate solution be 10 mu M, and incubating for 24 hours;
S2, treating according to the method in the example 4, but only incubating SNX3 antibody;
S3, incubating a fluorescent secondary antibody and staining by using DAPI according to the operation of the embodiment 3, and observing fluorescence distribution under a laser confocal microscope.
The experimental results are shown in FIG. 6, and it can be seen from the experimental results that green fluorescence is GFP-coupled compound LC-4, red fluorescence is sorting connexin 3, and co-localization yellow pixel and co-localization coefficient indicate that interaction exists between compound LC-4 and SNX 3.
Example 73 interaction of an oxo-benzofuran-based LC-4-GFP conjugate with SNX3
S1, taking well-grown rat myocardial cell strain H9C2, putting the well-grown rat myocardial cell strain H9C2 into two 10cm cell culture dishes, adding Ad-SNX3 virus to overexpress SNX3 when the cell density reaches 70% -80%, respectively adding 3 mu L of LC-4-GFP conjugate solution after 12 hours to make the final concentration of the strain 10 mu M, and incubating for 24 hours;
S2, taking about 20 mug of protein from total protein as an Input control sample before IP after cell protein cleavage, respectively adding a Flag tag antibody (Flag-tag) and a GFP tag antibody (GFP-tag), and slowly shaking at 4 ℃ overnight by taking the same kind of epidemic globulin G (IgG) as a negative control;
s3, preparing a WB sample and performing Western blot detection by the same operation as in the example 5.
The experimental results are shown in FIG. 7, and it can be seen from the experimental results that the compound LC-4 binds to SNX 3.
Example 83 Effect of the oxo-benzofurans LC-4 on cardiac fibrosis
S1, culturing SD (dairy mouse) primary myocardial fibroblasts by using a complete culture medium: high sugar DMEM+10% fetal bovine serum, and the culture condition is 5% carbon dioxide and 37 ℃;
S2, after the cells grow to 70% -80% of density, starving the cells for 12 hours by using a high-sugar DMEM culture medium containing 1% of fetal calf serum;
s3, after starvation treatment, replacing the culture medium with high-sugar DMEM containing 10% of fetal calf serum, and simultaneously pre-dosing compound LC-4 with different concentrations;
s4, after pre-dosing for 2 hours, 1 mu M angiotensin II (Ang II) is dosed for 48 hours to induce myocardial fibroblast fibrosis, and Ang II is added every 12 hours;
And S5, collecting cell proteins after 48 hours, and performing Western blot detection after quantitative determination by using BCA.
As shown in fig. 8, it can be seen from the experimental results that the individual Ang ii group fibrosis-related proteins Fibronectin (FN), collagen type I (COL 1 A1) and α -smooth muscle actin (α -SMA) were all significantly expressed and significantly down-regulated by administration of different concentrations of compound LC-4, and were dose dependent; the above results show that compound LC-4 can inhibit myocardial fibrosis induced by Ang ii.
Example 93 Effect of the oxo-benzofuran type LC-4 on ISO-induced heart failure mice cardiac function
40 Male 6-8 week old C57BL/6 mice were selected and randomly divided into 5 groups, namely 8 control groups, 8 model groups, 85 mg.kg -1 compound groups, 810 mg.kg -1 compound groups and 820 mg.kg -1 compound groups: compound LC-4 was administered intraperitoneally, and after 7 days of pre-administration, molding was performed; both the model group and the drug group were subcutaneously injected with isoprenaline (ISO, 5 mg.kg -1) once a day for a total of 10 days, simulating the chronic heart failure animal model, and the control group mice were given an equal volume of physiological saline.
The results of the echocardiography of the mice are shown as A in fig. 9, and further data analysis of the echocardiography results can be seen: compared with the control group, the parameters of the ISO group of mice such as ultrasonic cardiac ejection fraction (ejection fraction, EF), left chamber short axis shortening rate (fraction shortening, FS), cardiac Output (CO), stroke Volume (SV) and the like are obviously reduced, and the parameters of the left chamber anterior wall thickness (LVAW) and the left chamber posterior wall thickness (LVPW) are obvious, which indicate that the ISO group of mice have reduced cardiac function and successful molding. The EF, FS, CO, SV was significantly up-regulated after treatment with different concentrations of LC-4, suggesting that the LC-4 groups were able to effectively improve the reduction in ISO-induced Ejection Fraction (EF), left ventricular short axis reduction (FS), cardiac Output (CO), stroke Volume (SV). The results indicate that the compound LC-4 can significantly improve ISO-induced heart function abnormality, and the compound LC-4 is shown to significantly improve ISO-induced chronic heart failure and has better heart protection effect.
Example 10 Effect of 3-oxo-benzofuran based LC-4 on ISO-induced heart failure and cardiac fibrosis
The heart morphology, tissue HE staining, sirius red staining, wheat germ lectin staining (WGA) sections of each group of experimental animals in example 9 were analyzed, and Western blot detection was performed after partial tissue lysis.
As shown in the experimental results in FIG. 10, the A-B shows that the ISO group heart appearance is obviously increased, and the compound LC-4 can improve the cardiac hypertrophy caused by ISO; C-D shows that the weight-to-weight ratio (HW/BW) and the length-to-length ratio (HW/TL) of the ISO group are obviously increased, which indicates that abnormal cardiac function can be improved after the treatment of compounds LC-4 with different concentrations; E-G shows that the cell arrangement disorder, fibrosis increase and collagen fiber abnormal deposition of the ISO group can be obviously improved after the LC-4 treatment of compounds with different concentrations.
Further western blot experiments see fig. 11, which shows that the ISO group fibrosis related protein FN expression levels are significantly up-regulated and expression is significantly reduced after treatment with different concentrations of compound LC-4.
The above results indicate that LC-4 can improve to some extent heart failure and heart fibrosis induced by ISO.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. The application of a 3-oxo-benzofuran compound or a derivative thereof in preparing a medicament for treating heart failure related diseases is characterized in that the 3-oxo-benzofuran compound has the following structure:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
2. The use according to claim 1, wherein the pharmaceutically acceptable salt of the 3-oxo-benzofuran compound is a salt of the 3-oxo-benzofuran compound with an organic or inorganic base.
3. The use according to claim 2, characterized in that the organic base is triethylamine, pyridine, sodium tert-butoxide, potassium tert-butoxide, diisopropylamine, n-butyllithium, isobutyllithium, tert-butyllithium, sodium methoxide, sodium ethoxide or sodium amide.
4. Use according to claim 2, characterized in that the inorganic base is sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium thiosulfate, sodium hydroxide, lithium hydroxide, potassium hydroxide, ammonia or ammonium bicarbonate.
5. The use according to any one of claims 1to 4, wherein the heart failure related disease comprises coronary heart disease, myocardial infarction, angina pectoris, myocarditis or cardiac fibrosis.
6. The use according to claim 5, wherein the medicament is in the form of a tablet, capsule, pill, powder, granule, solution or injection.
7. A composition for treating a heart failure related disorder, the composition comprising a 3-oxo-benzofuran compound or derivative thereof and an anti-heart failure agent; the 3-oxo-benzofuran compounds have the following structures:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
8. The composition for treating heart failure-related diseases according to claim 7, wherein the anti-heart failure drug is selected from one or more of an angiotensin receptor enkephalinase inhibitor or an angiotensin converting enzyme inhibitor/angiotensin ii receptor antagonist, sodium-glucose cotransporter 2 inhibitor, beta blocker, mineralocorticoid receptor antagonist.
9. Use of a 3-oxo-benzofuran compound or derivative thereof in the preparation of an inhibitor of sortilin 3, wherein the 3-oxo-benzofuran compound has the following structure:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
10. Use of a 3-oxo-benzofuran compound or derivative thereof in the preparation of a transcriptional activator protein 3 phosphorylation inhibitor, wherein the 3-oxo-benzofuran compound has the following structure:
Wherein the derivative is pharmaceutically acceptable salt, precursor, chiral isomer, esterified substance or solvate of the 3-oxo-benzofuran compound.
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