Use of a compound targeting lif-lifr in the manufacture of a medicament for promoting tissue anti-fibrosis

Abstract

The present disclosure relates to the use of a compound targeting LIF-LIFR in the preparation of a medicine for promoting the anti-fibrosis of tissues. It also relates to a medicine for promoting the anti-fibrosis of tissues, the active ingredient of which comprises a compound targeting LIF-LIFR. The compound targeting LIF-LIFR provided by the present disclosure exhibits good anti-fibrosis activity; it can inhibit the expression of Col1a1 (COL1A1), Fn, Acta2, Lif (LIF), Lifr (LIFR), alpha-SMA and other proteins, thereby effectively preventing, improving or eliminating the fibrosis of organ tissues and its complications.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to the field of biological medicine, in particular, to the use of a compound targeting LIF-LIFR in the preparation of a drug for promoting tissue anti-fibrosis. BACKGROUND

[0002] Fibrosis can occur in various organs, the main pathological changes are the increase of fibrous connective tissue in organ tissue (such as lung tissue, liver tissue, myocardial tissue), the decrease of parenchymal cells, the continuous progression can cause the destruction of organ structure and the reduction of function, and even failure, which seriously threatens human health and life.

[0003] Anti-fibrosis can slow down the progression of fibrosis, promote the repair and / or regeneration of tissue, and delay the deterioration of organ function; it can also prevent the occurrence of fibrosis-related complications; thus, the quality of life and prognosis of patients are improved.

[0004] Therefore, the development of anti-fibrosis drugs is of great significance. SUMMARY

[0005] The purpose of the present disclosure is to provide the use of a compound targeting LIF-LIFR in the preparation of a drug for promoting tissue anti-fibrosis. The compound provided by the present disclosure exhibits good anti-fibrosis activity.

[0006] To achieve the above-mentioned purpose, the first aspect of the present disclosure provides the use of a compound targeting LIF-LIFR in the preparation of a drug for promoting tissue anti-fibrosis, wherein the compound is selected from any one of the compounds represented by formula (1), formula (2), formula (3) and formula (4):

[0007] Formula (1), Formula (2),

[0008] Formula (3), Formula (4).

[0009] Optionally, the promotion of tissue anti-fibrosis is the prevention, improvement or elimination of fibrosis of organ tissue and its complications.

[0010] Optionally, the organ includes any one or several of lung, liver, kidney, heart and skin.

[0011] Optionally, the fibrosis of organ tissue and its complications include idiopathic fibrosis, connective tissue disease-related interstitial lung disease, chronic fibrosing interstitial lung disease of the progressive phenotype, fatty liver fibrosis, alcoholic liver fibrosis, hypertensive heart disease, myocardial infarction, scleroderma, hypertrophic scar and keloid.

[0012] Optionally, the administration mode of the medicine comprises injection administration and / or oral administration.

[0013] Optionally, the administration amount of the compound is 20-40 mg / kg.

[0014] Optionally, the medicine further comprises an excipient, and the excipient comprises at least one of carboxymethyl cellulose, dimethyl sulfoxide and Tween-80.

[0015] The second aspect of the present disclosure provides a medicine for promoting tissue anti-fibrosis, and an active ingredient of the medicine comprises a compound targeting LIF-LIFR, and the compound is selected from any one of the compounds shown in formula (1), formula (2), formula (3) and formula (4):

[0016] Formula (1), Formula (2),

[0017] Formula (3), Formula (4);

[0018] Optionally, the medicine further comprises an excipient, and the excipient comprises at least one of carboxymethyl cellulose, dimethyl sulfoxide and Tween-80.

[0019] Through the above technical solution, the present disclosure provides the use of the compound targeting LIF-LIFR in the preparation of the medicine for promoting tissue anti-fibrosis. The compound targeting LIF-LIFR provided by the present disclosure shows good anti-fibrosis activity; can inhibit the expression of Col1a1 (COL1A1), Fn, Acta2, Lif (LIF), Lifr (LIFR), α-SMA and other proteins, thereby effectively preventing, improving or eliminating the fibrosis of organ tissues and its complications.

[0020] Other features and advantages of the present disclosure will be described in detail in the following specific embodiment part. BRIEF DESCRIPTION OF DRAWINGS

[0021] The accompanying drawings are included to provide a further understanding of the present disclosure and constitute a part of the specification, and are used together with the following specific embodiments to explain the present disclosure, but do not constitute a limitation on the present disclosure. In the drawings:

[0022] Figure 1 is a safety test result graph of the compounds shown in formula (1)-formula (4) in Example 2.

[0023] Figure 2 is a safety test result graph of the compounds shown in formula (5)-formula (9) in Example 2.

[0024] Figure 3is a graph showing the results of staining of lung tissue sections of bleomycin-induced pulmonary fibrosis in mice treated with the compound represented by formula (4) in Example 3.

[0025] Figure 4 is the effect of the compounds represented by formulae (1) to (4) in Example 4 and formulae (5) to (9) in Comparative Example 1 on the expression of fibrosis-related proteins by fibroblasts stimulated with TGF-β1.

[0026] Figure 5 is an electropherogram of the expression of fibrosis-related proteins by fibroblasts stimulated with LIF in Example 5.

[0027] Figure 6 is the effect of the compounds represented by formulae (1) to (4) in Example 5 on the expression of fibrosis-related proteins by fibroblasts stimulated with LIF.

[0028] Figure 7 is an electropherogram of the expression of fibrosis-related proteins by fibroblasts stimulated with LIF in Comparative Example 2.

[0029] Figure 8 is the effect of the compounds represented by formulae (5) to (9) in Comparative Example 2 on the expression of fibrosis-related proteins by fibroblasts stimulated with LIF.

[0030] Figures 9-12 is the effect of the compounds represented by formulae (1) to (4) in Example 6 on the expression of fibrosis-related proteins by tubular epithelial cells stimulated with TGF-β1.

[0031] Figure 13 is an electropherogram of the expression of fibrosis-related proteins by tubular epithelial cells stimulated with LIF in Example 7.

[0032] Figure 14 is the effect of the compounds represented by formulae (1) to (4) in Example 7 on the expression of fibrosis-related proteins by tubular epithelial cells stimulated with LIF.

[0033] Figure 15 is the effect of the compounds represented by formulae (1) to (4) in Example 8 on serum creatinine in renal interstitial fibrosis induced by the UIRI model. DETAILED DESCRIPTION

[0034] A detailed description of specific embodiments of the disclosure follows, in reference to the attached drawings. It should be understood that the detailed description and specific examples, while indicating preferred embodiments of the disclosure, are given by way of illustration only and are not intended to limit the disclosure.

[0035] The first aspect of the present disclosure provides use of a compound targeting LIF-LIFR in the preparation of a medicament for promoting anti-fibrosis of an organ, wherein the compound is selected from any one of the compounds represented by formula (1), formula (2), formula (3) and formula (4):

[0036] formula (1), formula (2),

[0037] formula (3), formula (4).

[0038] In the present disclosure, the inventors of the present disclosure surprisingly found that the compounds represented by formula (1), formula (2), formula (3) and formula (4) have high affinity with LIF-LIFR (leukemia inhibitory factor and its specific receptor) and exhibit good anti-fibrosis activity; specifically, the inventors of the present disclosure verified through experiments that the compounds represented by formula (1), formula (2), formula (3) and formula (4) can inhibit the expression of COL1A1, FN, a-SMA, LIF, LIFR and other proteins, thereby effectively preventing, improving and eliminating fibrosis of an organ and its complications.

[0039] In the present disclosure, the promotion of anti-fibrosis of an organ refers to prevention, improvement or elimination of fibrosis of an organ and its complications; in an embodiment of the present disclosure, the organ includes any one or several of lung, liver, kidney, heart and skin. Specifically, fibrosis of an organ and its complications can include idiopathic fibrosis, connective tissue disease-related interstitial lung disease, chronic fibrosing interstitial lung disease of the progressive phenotype, fatty liver fibrosis, alcoholic liver fibrosis, hypertensive heart disease, myocardial infarction, scleroderma, hypertrophic scars and keloids.

[0040] In an embodiment of the present disclosure, the administration mode of the medicament includes injection administration and / or oral administration.

[0041] In an embodiment of the present disclosure, the administration amount of the compound is 20-40 mg / kg. In the above embodiment, the administration amount of the compound refers to the administration amount in a mouse model of pulmonary fibrosis.

[0042] In an embodiment of the present disclosure, the medicament further includes an excipient, and the excipient includes at least one of carboxymethyl cellulose, dimethyl sulfoxide and Tween-80.

[0043] The second aspect of the present disclosure provides a medicament for promoting anti-fibrosis of an organ, wherein the active ingredient of the medicament includes a compound targeting LIF-LIFR, and the compound is selected from any one of the compounds represented by formula (1), formula (2), formula (3) and formula (4):

[0044] Formula (1), Formula (2),

[0045] Formula (3), Formula (4).

[0046] In an embodiment of the present disclosure, the medicine further comprises an excipient, and the excipient comprises at least one of carboxymethyl cellulose, dimethyl sulfoxide, and Tween-80.

[0047] The present application is further illustrated by the following examples, but the present application is not limited in any way by the following examples.

[0048] The compounds used in the following examples and comparative examples are shown in Table 1.

[0049] Table 1

[0050]

[0051]

[0052] Example 1

[0053] This example is used to illustrate the affinity test of the compounds shown in Formula (1) to Formula (9) to LIF (leukemia inhibitory factor).

[0054] The affinity test uses a surface plasmon resonance (SPR) instrument (model Biacore 8K), and the specific steps include:

[0055] The system temperature is set to 25℃, the flow rate is set to 10 μL / min, and the 1×PBS-P buffer is passed until the baseline is stable. A CM5 chip is selected, and the coupling of LIFR is operated according to the NHS / EDC coupling method, and the coupling steps are as follows: (a) Activation: 900 s of sample injection at a flow rate of 10 μL / min, and a mixture of NHS / EDC (0.1M NHS and 0.4M EDC, mixed in a volume ratio of 1:1 before use, and used immediately) is used to activate the carboxyl groups of the dextran on the chip surface; (b) Immobilization: a solution of LIFR at 20 ng / μL is prepared using a sodium acetate buffer (pH 5.5), and 1200 s of sample injection is performed at a flow rate of 10 μL / min; (c) Blocking: 900 s of blocking is performed using 1M ethanolamine hydrochloride, and the coupling process is completed.

[0056] The compound to be detected was dissolved in PBS-P buffer (1.0x PBS, pH 7.4, 0.15 M NaCl, 0.05% tween-20) to prepare a concentration gradient of 0.01 μM, 0.0195 μM, 0.039 μM, 0.078 μM, 0.156 μM, 0.3125 μM, 0.625 μM, 1.25 μM, and EC359 was used as a positive control for kinetic constant analysis. The results are shown in Table 2.

[0057] Table 2

[0058]

[0059] As shown in Table 2, the compounds represented by formula (1) to formula (8) have an affinity for LIF comparable to that of the positive control; indicating that the compounds represented by formula (1) to formula (8) have different degrees of binding affinity for LIF.

[0060] Example 2

[0061] This example is used to illustrate the in vitro safety (cytotoxicity) test of the compounds represented by formula (1) to formula (9).

[0062] The in vitro safety test uses the CCK-8 method to detect NRK-49F cytotoxicity, and the specific steps include:

[0063] NRK-49F cells were seeded in a 96-well plate at a density of 5x10 4 cells / well, and 100 μL of DMEM / F12 (containing 10% FBS and 1% double antibody) cell culture solution was added to each well. After incubation in a 37°C cell culture incubator for 24 h, the culture solution was replaced with DMEM / F12 (containing double antibody) cell culture solution containing 1.5 ng / mL, 3 ng / mL, 6 ng / L, 12.5 ng / mL, 25 ng / mL, 50 ng / mL, and 100 ng / mL of the compound represented by formula (1) prepared in advance, and incubation was continued for 48 h. The old culture solution was carefully removed and replaced with fresh DMEM / F12 (containing 1% double antibody) cell culture solution, and 10 μL of CCK-8 solution was added to each well, which was incubated at 37°C for 0.5 h. The OD 450nm was read using a microplate reader, and the relative cell survival rate was calculated.

[0064] The compounds represented by formula (2) to formula (9) were tested for cytotoxicity according to the above method, and the results are shown in Figure 1 and 2 .

[0065] From Figure 1 and Figure 2It can be seen that the compound shown in formula (6) is highly toxic, while the compounds shown in formula (1), formula (2), formula (3), formula (4), formula (5), formula (7), formula (8) and formula (9) have better safety.

[0066] Example 3

[0067] This example illustrates that the compound shown in formula (4) can prevent the progression of bleomycin-induced pulmonary fibrosis in mice.

[0068] (1) The methods for preparing a bleomycin-induced mouse pulmonary fibrosis model include:

[0069] Healthy male C57BL / 6J mice aged 5 weeks with an average weight of 20 g were selected and acclimatized for 7 days. Then, a physiological saline solution containing bleomycin was injected intratracheally at a dose of 2.5 mg / kg / day. The model was successfully established when the mice exhibited huddling and lying down, weight loss, and inflammatory infiltration and collagen deposition in the lung tissue pathology examination on the 7th day of culture.

[0070] (2) Feeding methods for bleomycin-induced mouse pulmonary fibrosis model:

[0071] Model group: The group model constructed in step (1) is divided into groups and fed with the compound shown in formula (4) at a rate of 30 mg / kg / day.

[0072] Blank control group: fed the same amount of blank solvent (physiological saline) as the model group.

[0073] Solvent control group (sham surgery group): fed the same amount of blank solvent (physiological saline) as the model group.

[0074] (3) Lung tissue was taken 21 days later for Masson staining to verify the progression of pulmonary fibrosis. The results are as follows: Figure 3 As shown, by Figure 3 It can be seen that formula (4) can significantly inhibit the progression of pulmonary fibrosis.

[0075] Example 4

[0076] This example illustrates that the compounds shown in formulas (1) to (4) can inhibit the expression of fibrosis-related proteins in rat kidney fibroblasts (NRK-49F) stimulated by TGF-β1.

[0077] The specific method is as follows:

[0078] NRK-49F cells were inoculated in DMEM / F12 mixed medium containing 10% FBS and 1% double antibody, and cultured in a 5% CO2 incubator. NRK-49F cells cultured for 24 h were divided into a negative control group (mixed culture with sterile PBS buffer), a positive control group (mixed culture with only 8 ng / mL human recombinant TGF-β1 protein (rhTGF-β1)), and four drug administration groups (mixed culture with 50 ng / mL compounds represented by formula (1) to formula (4) and 8 ng / mL rhTGF-β1, respectively). The four drug administration groups were replaced with DMEM / F12 (containing 1% double antibody) cell culture solution containing 50 ng / mL compounds represented by formula (1) to formula (4) configured in advance 2 h in advance. After 2 h, the positive control group and the drug administration group were added with 8 ng / mL of rhTGF-β1, and continued to be cultured for 48 h. The supernatant was discarded, the cells were washed with PBS for 3 times, the cells were collected, the mRNA was extracted, and the mRNA expression level of fibrosis-related proteins in the extracellular matrix of NRK-49F cells was detected by RT-qPCR. The results are shown in Table 1. Figure 4

[0079] Comparative Example 1

[0080] The same method as in Example 4 was used, except that five drug administration groups were set up, and the cultured NRK-49F cells were mixed with 10 ng / mL compound represented by formula (6), 50 ng / mL compound represented by formula (5), and compounds represented by formula (7) to formula (9), respectively. The remaining steps were the same as in Example 4, and the results are shown in Table 2. Figure 4

[0081] From Table 2, it can be seen that the compounds represented by formula (1), formula (3), and formula (4) can effectively inhibit the expression of Figure 4 in the extracellular matrix of NRK-49F, thereby effectively improving the fibrosis caused by TGF-β1 stimulating NRK-49F cells. Col1a1, Fn, Acta2, Lif, Lifr Example 5

[0082] This example is used to illustrate that the compounds represented by formula (1) to formula (4) can inhibit the expression of fibrosis-related proteins in rat kidney fibroblasts (NRK-49F) stimulated by LIF.

[0083] The specific method is as follows:

[0084]

[0085] ​​​NRK-49F cells were inoculated in DMEM / F12 mixed medium containing 10% FBS and 1% double antibody and cultured in a 5% CO2 incubator. NRK-49F cells cultured for 24 h were divided into a negative control group (mixed with sterile PBS buffer for culture), a positive control group (mixed with 12 ng / mL human recombinant LIF protein (rhLIF) for culture only), and four drug administration groups (mixed with 1, 5, 10, 20, 50 ng / mL compounds represented by formula (1) to formula (4) and 12 ng / mL rhLIF, respectively). The four drug administration groups were replaced with DMEM / F12 (containing 1% double antibody) cell culture solution containing 1, 5, 10, 20, 50 ng / mL compounds represented by formula (1) to formula (4) configured in advance 2 h in advance, and 12 ng / mL rhLIF was added to the positive control group and the drug administration group 2 h later. Continue to culture for 48 h, discard the supernatant, wash the cells with PBS for 3 times, collect the cells, extract the protein, and use Western Blot to detect the expression of fibrosis-related proteins in the extracellular matrix of NRK-49F cells. The results are shown in Figures 5-6 .

[0086] Comparative Example 2

[0087] The same method as in Example 5 was used, except that the drug administration groups of the compounds represented by formula (5) to formula (9) were set up, and the NRK-49F cells after culture were mixed with 1, 5, 10 ng / mL formula (6), 1, 5, 10, 20, 50 ng / mL compounds represented by formula (5), formula (7) to formula (9) for culture, and the remaining steps were the same as in Example 5. The results are shown in Figures 7-8 .

[0088] From Figures 5-8 It can be seen that, compared with formula (5) to formula (9), the compounds represented by formula (1), formula (2), formula (3) and formula (4) can effectively inhibit the expression of COL1A1, LIFR, α-SMA and LIF, thereby effectively improving the fibrosis caused by LIF stimulating NRK-49F cells.

[0089] Example 6

[0090] This example is used to illustrate that the compounds represented by formula (1) to formula (4) can inhibit the expression of fibrosis-related proteins in mouse primary renal tubular epithelial cells stimulated by TGF-β1.

[0091] The specific method is:

[0092] Eight-week-old C57BL / 6 male mice were euthanized with sodium pentobarbital. Kidneys were removed in a clean biosafety cabinet, rinsed with sterile, ice-cold physiological saline to remove blood cells, minced with ophthalmic scissors, and then digested with type IV collagenase at 37°C until a paste-like consistency was achieved. Digestion was terminated by adding PBS buffer containing 10% FBS. The cells were centrifuged at 500 rpm for 2 minutes at 4°C, the supernatant was discarded, and the pellet was resuspended in 5 mL of 32% Percoll solution. The cells were centrifuged at 2000g at 4°C for 10 minutes, with a slow deceleration (increase rate 9, decrease rate 5). The supernatant was discarded, and the cells were resuspended in DMEM / F12 (containing 1% penicillin and antibiotics), centrifuged at 500 rpm for 2 minutes at 4°C. The supernatant was discarded, and the cells were resuspended in DMEM / F12 cell culture medium containing 10% FBS and 1% penicillin and antibiotics. The resuspended solution was transferred to culture dishes and cultured at 37°C for subsequent experiments.

[0093] Once the cells reached a suitable density, mouse primary renal tubular epithelial cells were divided into a negative control group (cultured with sterile PBS buffer), a positive control group (cultured with only 8 ng / mL rhTGF-β1), and four drug-treated groups (cultured with 1, 5, 10, 20, and 50 ng / mL of the compounds shown in formulas (1)-(4) and 8 ng / mL rhTGF-β1, respectively). Two hours before treatment, the four drug-treated groups were replaced with DMEM / F12 (containing 1% penicillin antibody) cell culture medium containing the compounds shown in formulas (1)-(4) at concentrations of 1, 5, 10, 20, and 50 ng / mL. Two hours later, 8 ng / mL rhTGF-β1 was added to the positive control group and the drug-treated groups, and the cells were cultured for another 48 hours. The supernatant was discarded, the cells were washed three times with PBS buffer, and the cells were collected. mRNA was extracted, and the mRNA expression level of fibrosis-related proteins in the extracellular matrix of mouse primary renal tubular epithelial cells was determined by RT-qPCR. The results are as follows: Figures 9-12 As shown.

[0094] Depend on Figures 9-12 It can be seen that the compounds shown in formulas (1), (2), (3), and (4) can effectively inhibit Col1a1, Fn, Acta2, Lif, Lifr The expression of [the substance] can effectively improve fibrosis caused by TGF-β1 stimulation of renal tubular epithelial cells.

[0095] Example 7

[0096] This embodiment is used to illustrate that the compounds shown in formulas (1)-(4) can inhibit the expression of fibrosis-related proteins in renal tubular epithelial cells stimulated by LIF.

[0097] The specific method is as follows:

[0098] Eight-week-old C57BL / 6 male mice were euthanized with sodium pentobarbital. Kidneys were removed in a clean biosafety cabinet, rinsed with sterile, ice-cold physiological saline to remove blood cells, minced with ophthalmic scissors, and then digested with type IV collagenase at 37°C until a paste-like consistency was achieved. Digestion was terminated by adding PBS buffer containing 10% FBS. The cells were centrifuged at 500 rpm for 2 minutes at 4°C, the supernatant was discarded, and the pellet was resuspended in 5 mL of 32% Percoll solution. The cells were centrifuged at 2000g at 4°C for 10 minutes, with a slow deceleration (increase rate 9, decrease rate 5). The supernatant was discarded, and the cells were resuspended in DMEM / F12 (containing 1% penicillin and antibiotics), centrifuged at 500 rpm for 2 minutes at 4°C. The supernatant was discarded, and the cells were resuspended in DMEM / F12 cell culture medium containing 10% FBS and 1% penicillin and antibiotics. The resuspended solution was transferred to culture dishes and cultured at 37°C for subsequent experiments.

[0099] Once the cells reached a suitable density, primary mouse renal tubular epithelial cells were divided into a negative control group (cultured with sterile PBS buffer), a positive control group (cultured with 12 ng / mL rhLIF only), and four drug-treated groups (cultured with compounds of formulas (1)-(4) at concentrations of 1, 5, 10, 20, and 50 ng / mL and 12 ng / mL rhLIF, respectively). Two hours prior to treatment, the four drug-treated groups were replaced with pre-prepared DMEM / F12 (containing 1% penicillin antibody) cell culture medium containing compounds of formulas (1)-(4) at concentrations of 1, 5, 10, 20, and 50 ng / mL. Two hours later, the positive control group and the drug-treated groups were treated with 12 ng / mL rhLIF and cultured for another 48 hours. The supernatant was discarded, and the cells were washed three times with PBS. Cells were collected, proteins were extracted, and the expression of fibrosis-related proteins in the extracellular matrix of renal tubular epithelial cells was detected using Western blotting. The results are as follows: Figures 13-14 As shown.

[0100] Depend on Figures 13-14 It can be seen that the compounds shown in formulas (1), (2), (3) and (4) can effectively inhibit the expression of COL1A1, LIFR, α-SMA and LIF, thereby effectively improving fibrosis caused by TGF-β1 stimulation of renal tubular epithelial cells.

[0101] Example 8

[0102] This embodiment is used to illustrate that the compounds shown in formulas (1)-(4) can inhibit renal interstitial fibrosis induced by the UIRI model.

[0103] The specific method is as follows:

[0104] 8w C57BL / 6 male mice were anesthetized with sodium pentobarbital, the abdomen was opened along the midline, the left kidney and renal artery were exposed, the left renal artery was clamped with a vascular clamp, the mouse was placed on a preheated 38℃ metal bath, physiological saline was dropped to the abdomen to prevent excessive evaporation of the mouse's body fluid, after 35 minutes, the mouse was taken out, the left kidney had turned purple black indicating that the modeling was successful, the vascular clamp was removed, the mouse's abdominal wound was sutured using single interrupted suture method, and the layers were sutured. After suture, the wound was wiped with 75% alcohol. Return to 37℃ constant temperature metal bath, wait for the mouse to wake up and return to the mouse cage. The sham group was treated with the same abdominal wound, then sutured, and not treated otherwise. Five days after the operation, the UIRI group was divided into 3 groups (UIRI+vehicle group, UIRI+EC330 group and UIRI+formula (1) group), the UIRI+vehicle group was given blank control reagent, the UIRI+EC330 group was injected with EC330 (5mg / kg) intraperitoneally, and the UIRI+formula (1) group was given formula (1) (9mg / kg) by gavage, for five consecutive days. On the 10th day after the operation, the mouse was anesthetized with sodium pentobarbital, the right kidney was opened dorsally, the right kidney and renal artery were exposed, the renal artery and ureter were tied tightly, and the mouse's right healthy kidney was removed. The mouse's dorsal wound was sutured using single interrupted suture method, and the layers were sutured. After suture, the wound was wiped with 75% alcohol. Return to 37℃ constant temperature metal bath, wait for the mouse to wake up and return to the mouse cage. On the 11th day after the operation, the mouse was sacrificed, the sample was taken, and the sample was detected. The results are shown in Figure 15 .

[0105] The same method as formula (1) was used, except that the concentration of formula (2) was 30mg / kg, the concentration of formula (3) was 3mg / kg, and the concentration of formula (4) was 30mg / kg, and the remaining steps were the same as example 8. The results are shown in Figure 15 .

[0106] As can be seen from Figure 15 , the compounds represented by formula (1), formula (2), formula (3) and formula (4) can effectively inhibit the increase of serum creatinine induced by the UIRI model, thereby effectively improving the renal interstitial fibrosis induced by the UIRI model in mice.

[0107] Examples 3-8 verify that the compounds represented by formula (1), formula (2), formula (3) and formula (4) have good anti-fibrosis effect.

[0108] The preferred embodiments of the present disclosure are described in detail above in combination with the drawings, but the present disclosure is not limited to the specific details in the above-described embodiments. Within the technical concept of the present disclosure, various simple modifications can be made to the technical solutions of the present disclosure, and these simple modifications all belong to the protection scope of the present disclosure.

[0109] It should be further noted that various specific technical features described in the above specific embodiments can be combined in any suitable manner, and the disclosure will not be repeated here for various possible combinations.

[0110] In addition, various different embodiments of the disclosure can also be combined with each other as long as they do not contradict the idea of the disclosure, and they should also be considered as disclosed by the disclosure.

Claims

1. Use of a compound targeting LIF-LIFR for the manufacture of a medicament for preventing, ameliorating or eliminating fibrosis of the kidney, characterized in that, The compound is selected from any one of compounds shown in formula (1), formula (2), formula (3) and formula (4): Formula (1), Formula (2), Formula (3), Formula (4).

2. Use according to claim 1, wherein, The administration mode of the medicine comprises injection administration and / or oral administration.

3. Use according to claim 2, wherein, The administration amount of the compound is 20-40 mg / kg.

4. The use according to claim 1, wherein, The medicine further comprises an excipient, and the excipient comprises at least one of carboxymethyl cellulose, dimethyl sulfoxide and Tween-80.

5. Use of a compound targeting LIF-LIFR for the manufacture of a medicament for preventing, ameliorating or eliminating fibrosis of the lung, characterized in that, The compound is the compound shown in formula (4): Formula (4).

6. Use according to claim 5, wherein, The pulmonary fibrosis comprises idiopathic pulmonary fibrosis, connective tissue disease-related interstitial lung disease and chronic fibrotic interstitial lung disease of progressive phenotype.

7. Use according to claim 5, wherein, The administration mode of the medicine comprises injection administration and / or oral administration.

8. Use according to claim 7, wherein, The administration amount of the compound is 20-40 mg / kg.

9. The use according to claim 5, wherein, The medicine further comprises an excipient, and the excipient comprises at least one of carboxymethyl cellulose, dimethyl sulfoxide and Tween-80.

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