Use of targeted inhibition of p300 / CBP in treating liver injury diseases

Abstract

Use of targeted inhibition of p300 / CBP in treating liver injury diseases, and a method for preventing and / or treating liver injury diseases by targeted inhibition of p300 / CBP in a subject in need thereof, the liver injury diseases comprising acute liver injury, chronic liver injury and / or liver failure, and acute-on-chronic liver failure. A p300 / CBP inhibitor has broad market application prospects in the treatment of acute and chronic liver injury and / or liver failure and acute-on-chronic liver failure.

Description

Application of targeted inhibition of p300 / CBP in the treatment of liver damage diseases Technical Field

[0001] The present invention belongs to the field of biomedicine technology and relates to the application of targeted inhibition of p300 / CBP in liver damage diseases. Specifically, it relates to the application of targeted inhibition of p300 / CBP in liver damage diseases, including acute and chronic liver damage and / or liver failure and acute-on-chronic liver failure. Background Art

[0002] The liver is a vital organ for metabolism and detoxification, and its functions are extremely important. However, in certain conditions, such as alcoholism, drug poisoning, viral infection, and autoimmune liver disease, the liver may suffer varying degrees of damage, leading to liver dysfunction and even acute liver failure, chronic liver damage, and acute-on-chronic liver failure.

[0003] Acute liver failure (ALF), a severe form of acute liver injury, occurs in a relatively short period of time (usually 1-26 weeks) and is characterized by a rapid deterioration of liver function over a period of days to weeks. Patients typically experience severe liver dysfunction, including jaundice (a symptom of yellowing of the skin and mucous membranes), coagulopathy (such as bleeding tendency), high bilirubin and elevated serum ammonia levels. The condition of patients with acute liver failure often worsens within a short period of time, sometimes even leading to coma and multiple organ dysfunction within a few days.

[0004] Chronic liver injury (CLI) refers to long-term liver damage or inflammation of varying degrees, typically developing slowly over months or years and potentially accompanied by a gradual impairment of liver function. Chronic liver injury typically presents no acute symptoms but gradually leads to fibrosis (liver fibrosis) and scarring (cirrhosis) of the liver tissue. If left untreated or uncontrolled, chronic liver injury can ultimately lead to serious complications such as liver dysfunction, portal hypertension, and hepatocellular carcinoma.

[0005] Acute-on-chronic liver failure (ACLF) is a serious liver disease that occurs suddenly on top of chronic liver disease. It manifests with severe symptoms such as jaundice, coagulopathy, and hepatic encephalopathy, often leading to multi-organ failure and death. When the disease occurs, patients often require emergency treatment.

[0006] Currently, there are no FDA-approved appropriate symptomatic drugs for acute and chronic liver injury and acute-on-chronic liver failure. Clinical treatment mainly relies on symptomatic treatment and supportive treatment, such as nutritional support and liver protection. However, these treatments cannot fundamentally improve liver damage and can only relieve patients' symptoms. Liver transplantation is a clinical treatment option with good efficacy. However, due to the shortage of donors, the difficulty of surgery, and high costs, this treatment method imposes a huge physical and psychological burden on patients. Clinical drug treatments for acute and chronic liver injury and acute-on-chronic liver failure mainly include the following aspects: liver protection drugs (such as alanine, glutathione, etc.), immunomodulatory drugs, antiviral drugs (if viral infection is the main cause of liver function deterioration), supportive treatment drugs (vitamins, mineral supplements), etc. These treatments cannot fundamentally improve liver damage and can only relieve patients' symptoms.

[0007] Inflammatory immune dysregulation plays a key role in acute liver failure, chronic liver injury, and acute-on-chronic liver failure. This commonality involves multiple complex biological processes, including abnormal activation of immune cells, excessive release of inflammatory mediators, immune cell infiltration leading to local inflammatory responses, and oxidative stress. Specifically, in acute liver failure, inflammatory immune dysregulation can often manifest as: a systemic, large-scale inflammatory response due to severe damage to liver cells; immune cells release large amounts of inflammatory mediators, such as TNF-α and IL-6, leading to exacerbated inflammatory damage; acute liver failure may cause the immune system's tolerance mechanism to fail, causing the immune system to attack its own tissues and aggravate liver damage. Immune dysregulation in chronic liver injury is often manifested as: persistent low-grade inflammation, leading to progressive liver fibrosis and cirrhosis; some immune cells continue to be activated during long-term liver inflammation, releasing inflammatory mediators, leading to liver tissue damage; the immune system may produce an abnormal immune response to liver tissue, leading to the development of autoimmune liver disease. The situation is even more complicated in acute-on-chronic liver failure, which combines the characteristics of acute and chronic liver damage. Immune dysregulation usually manifests as the characteristics of acute liver failure, but patients already have a background of chronic liver damage, so it is often accompanied by a significant systemic inflammatory response (immune cell activation and the release of large amounts of inflammatory mediators). At the same time, immunosuppression occurs and may cause the body to be unable to effectively respond to infection, increasing the risk of infectious complications. Infection is one of the most common causes of ACLF deterioration.

[0008] Considering that inflammatory immune dysregulation plays an important role in the pathogenesis of acute and chronic liver injury and acute-on-chronic liver failure, understanding this commonality will help us better understand the pathogenesis of these liver diseases and also provide important clues for developing treatment strategies, such as ameliorating liver damage by regulating immune responses.

[0009] CBP and p300 belong to the same protein family and share high similarity in their structural motifs. They catalyze the acetylation of H3K18 and H3K27, respectively, and their genes are expressed in all mammalian cells. The field of epigenetics, to which p300 / CBP proteins belong, is currently a hot topic of research. p300 / CBP proteins are directly linked to processes such as immune cell proliferation, differentiation, and function. Furthermore, targeted inhibition of p300 / CBP has recently been reported to modulate immune responses and suppress the development and progression of various inflammatory conditions, such as rheumatoid arthritis. p300 / CBP inhibitors are a class of epigenetic drugs that inhibit the activity of CBP (CREB-binding protein) and p300 proteins, proteins that play important roles in regulating the transcription and expression of inflammatory immune genes. Therefore, targeted inhibition of p300 / CBP has shown promising results in the treatment of various inflammatory and immune-related diseases.

[0010] Currently, there is a lack of effective treatments for acute liver injury or liver failure, chronic liver injury, and acute-on-chronic liver failure on the market. Therefore, finding an effective treatment is of great clinical significance for improving patient survival and therapeutic outcomes.

[0011] Summary of the Invention

[0012] The present invention aims to at least partially address one of the technical problems in the related art. To this end, one object of the present invention is to provide a new target and related methods for preventing and / or treating acute and chronic liver damage diseases and acute-on-chronic liver failure.

[0013] The present invention is based on the following findings:

[0014] p300 / CBP inhibitors can significantly inhibit the secretion of inflammatory factors by immune cells; in an acute liver injury mouse model, they improve the survival rate of mice, reduce the transcription level of inflammatory factors in mouse liver tissue, improve liver tissue damage and reduce liver cell apoptosis; in a chronic liver injury mouse model, they reduce the expression of liver tissue fibrosis factors and the formation of liver tissue fibrous cords; reduce the levels of liver enzymes ALT and AST, etc.; in a chronic liver failure mouse model, they significantly delay the peak of death of animals with chronic liver failure and increase the survival rate of animals, and improve liver and kidney blood flow disorders.

[0015] To this end, according to a first aspect of the present invention, the present invention provides a method for preventing and / or treating liver damage diseases by targeted inhibition of p300 / CBP in a subject in need thereof.

[0016] In a specific embodiment, the targeted inhibition of p300 / CBP includes targeting the HAT domain and BRD domain of p300 / CBPp300p300.

[0017] In a specific embodiment, the p300 treatment or prevention includes producing one or more of the following effects on the subject: inhibiting the secretion of inflammatory factors by immune cells, improving liver tissue damage, reducing hepatocyte apoptosis, lowering liver enzyme levels, improving liver and kidney blood flow disorders, and improving survival rate.

[0018] In a specific embodiment, the targeted inhibition of p300 / CBP comprises administering to the subject a therapeutically effective amount of a p300 / CBP inhibitor, including but not limited to CCS1477, GNE207, NEO2734, A-485, and CBP30. Those skilled in the art can select any suitable p300 / CBP inhibitor as needed to complete the present invention, and it is within the scope of protection of the present invention.

[0019] In a specific embodiment, the liver injury disease includes acute liver injury, chronic liver injury and / or liver failure and acute-on-chronic liver failure.

[0020] In a specific embodiment, the liver injury disease is acute liver injury, and the p300 / CBP inhibitor is administered to the subject at a dosage of 20-40 mg / Kg for a single administration, for example, the drug is administered to the subject at a dosage of 20 mg / Kg, 25 mg / Kg, 30 mg / Kg, 35 mg / Kg and 40 mg / Kg for a single administration.

[0021] In a specific embodiment, the liver injury disease is chronic liver injury, and the p300 / CBP inhibitor is administered to the subject at a dose of 3-10 mg / Kg·day, for example, 5 mg / Kg·day.

[0022] In a specific embodiment, the liver damage disease is acute-on-chronic liver failure, and the dosage of the p300 / CBP inhibitor administered to the subject is a single administration of 5-20 mg / Kg, for example, the dosage of the drug administered to the subject is a single administration of 5 mg / Kg, 10 mg / Kg, 15 mg / Kg and 20 mg / Kg.

[0023] According to the second aspect of the present invention, the present invention further provides a use of a p300 / CBP inhibitor in the preparation of a medicament for preventing and / or treating liver damage diseases.

[0024] In a specific embodiment, the p300 / CBP inhibitor targets the HAT domain and the BRD domain of p300 / CBP.

[0025] In a specific embodiment, the liver injury disease includes acute liver injury, chronic liver injury and / or liver failure and acute-on-chronic liver failure.

[0026] In a specific embodiment, the p300 / CBP inhibitor includes but is not limited to CCS1477, GNE207, NEO2734, A-485 and CBP30.

[0027] According to the third aspect of the present invention, the present invention further provides a drug for preventing and / or treating liver damage diseases, wherein the active ingredient of the drug comprises a p300 / CBP inhibitor.

[0028] In a specific embodiment of the present invention, the p300 / CBP inhibitor includes a compound targeting the HAT domain and BRD domain of p300 / CBP.

[0029] In a specific embodiment of the present invention, the p300 / CBP inhibitors include but are not limited to CCS1477, GNE207, NEO2734, A-485 and CBP30.

[0030] In a specific embodiment of the present invention, the liver injury disease includes acute liver injury, chronic liver injury and / or liver failure and acute-on-chronic liver failure.

[0031] In a specific embodiment of the present invention, the dosage form of the drug includes but is not limited to injection, tablets, capsules, oral granules, and enema. The drug of the present invention can be prepared into any dosage form that is convenient for administration.

[0032] According to the fourth aspect of the present invention, the present invention also proposes a pharmaceutical composition for preventing and / or treating liver damage diseases, which comprises the drug described above and a pharmaceutically acceptable excipient, wherein the excipient is one or any combination selected from a binder, a filler, a coating polymer, a plasticizer, a glidant, a disintegrant and a lubricant.

[0033] However, it should be understood that any mode of administration is within the scope of protection of the present invention.

[0034] On the basis of conforming to the common sense in this field, the above-mentioned preferred conditions can be arbitrarily combined without exceeding the concept and protection scope of the present invention. Beneficial effects

[0035] The inventors of this application discovered that slowing down the release of inflammatory factors and regulating immune responses are key points and directions in the study of acute-on-chronic liver failure. CBP / p300 inhibitors are a class of epigenetic drugs that can inhibit the activity of CBP (CREB-binding protein) and p300 proteins, which play an important role in the transcription and expression regulation of inflammatory immune genes.

[0036] The inventors of this application have also unexpectedly discovered that when a p300 / CBP inhibitor is selected from CCS1477, GNE207, NEO2734, A-485, and CBP30, and particularly when the p300 / CBP inhibitor is CCS1477, it can effectively prevent or treat liver damage, successfully protecting liver cells, reducing liver cell death caused by damage, reducing the release of inflammatory factors from immune cells, improving blood flow disorders in the liver and kidneys, delaying the peak of mortality in animals with acute-on-chronic liver failure, and improving the survival rate of individual animals. Furthermore, the aforementioned p300 / CBP inhibitors proposed in this invention are all small molecule compounds with low preparation, transportation, and storage costs, making them suitable for timely application in the event of acute liver damage or failure and acute-on-chronic liver failure.

[0037] In summary, the targeted inhibition of p300 / CBP proposed in this application can not only effectively solve the problem of the lack of ideal drugs and therapeutic methods for acute-on-chronic liver failure, but also effectively prevent or treat liver damage diseases by inhibiting the secretion of inflammatory factors of individual immune cells, improving liver tissue damage, reducing hepatocyte apoptosis, lowering liver enzyme levels, and improving animal survival rate. Therefore, the method of targeted inhibition of p300 / CBP proposed in this application has broad market application prospects in the treatment of acute and chronic liver injury and / or liver failure and acute-on-chronic liver failure. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] FIG1 is a graph comparing the mRNA levels of inflammatory factors in cells in a control group after LPS stimulation and a treatment group treated with different concentrations of p300 / CBP inhibitors 2 hours before LPS stimulation according to Example 1 of the present invention.

[0039] FIG2 is a graph comparing the levels of inflammatory cytokines in the control group after LPS stimulation and the treatment group treated with different concentrations of p300 / CBP inhibitors 2 hours before LPS stimulation according to Example 1 of the present invention.

[0040] Figure 3 is a graph comparing the survival of mice in the LPS-induced acute liver failure model in Example 2 of the present invention, using a vehicle and LPS / D-GalN injection modeling group, a treatment group that received a single intraperitoneal injection of CCS1477 4 hours before LPS / D-GalN injection, and a group that received only a single intraperitoneal injection of CCS1477 to detect drug toxicity.

[0041] Figure 4 is a comparison of the gross conditions of the liver tissues of mice in the LPS-induced acute liver failure model in Example 2 of the present invention, using solvent and LPS / D-GalN injection modeling groups and the treatment group given a single intraperitoneal injection of CCS1477 4 hours before LPS / D-GalN injection.

[0042] Figure 5 is a comparison of hematoxylin-eosin staining of liver tissues of mice in the LPS-induced acute liver failure model in Example 2 of the present invention, using solvent and LPS / D-GalN injection modeling groups and the treatment group given a single intraperitoneal injection of CCS1477 4 hours before LPS / D-GalN injection.

[0043] Figure 6 is a comparison of TUNEL (terminal dexynucleotidyl transferase-mediated dUTP nick end labeling) apoptosis staining in liver tissue of mice in the LPS-induced acute liver failure model using vehicle and LPS / D-GalN injection groups and the treatment group given a single intraperitoneal injection of CCS1477 4 hours before LPS / D-GalN injection according to Example 2 of the present invention.

[0044] Figure 7 is a comparison of serum biochemical indicators of hepatocellular damage: aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the LPS-induced acute liver failure model in mice according to Example 2 of the present invention, using a vehicle and LPS / D-GalN injection modeling group, a treatment group given a single intraperitoneal injection of CCS1477 4 hours before LPS / D-GalN injection, and a group only given a single intraperitoneal injection of CCS1477 to detect drug toxicity.

[0045] Figure 8 is a comparison of the mRNA levels of inflammatory factors in the liver tissue of mice in the LPS-induced acute liver failure model in Example 2 of the present invention, using solvent and LPS / D-GalN injection modeling groups and the treatment group given a single intraperitoneal injection of CCS1477 4 hours before LPS / D-GalN injection.

[0046] Figure 9 is a comparison of the gross conditions of the liver tissues of mice in the vehicle control group, the CCl4 injection modeling group, and the CCl4 injection modeling and CCS1477 intraperitoneal injection treatment group according to Example 3 of the present invention, a chronic mouse liver fibrosis injury model induced by CCl4.

[0047] Figure 10 is a comparison of Sirius red staining of liver tissues of mice in the CCl4-induced chronic mouse liver fibrosis injury model according to Example 3 of the present invention, a vehicle control group, a CCl4 injection modeling group, and a CCl4 injection modeling group simultaneously given CCS1477 intraperitoneal injection treatment group.

[0048] Figure 11 is a comparison of α-SMA and Col1a1 staining of liver tissues of mice in the CCl4-induced chronic liver fibrosis injury model in Example 3 of the present invention, including the vehicle control group, the CCl4 injection modeling group, and the CCl4 injection modeling and concurrent CCS1477 intraperitoneal injection treatment group.

[0049] Figure 12 is a comparison chart of the serum biochemical indicators of liver cell damage: aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the vehicle control group, the CCl4 injection modeling group and the CCl4 injection modeling and CCS1477 intraperitoneal injection treatment group according to Example 3 of the present invention, a chronic mouse liver fibrosis injury model induced by CCl4.

[0050] Figure 13 is a comparison of the mRNA levels of liver tissue fibrosis indicators in the CCl4-induced chronic mouse liver fibrosis injury model according to Example 3 of the present invention, a vehicle control group, a CCl4 injection modeling group, and a CCl4 injection modeling group simultaneously given intraperitoneal injection of CCS1477.

[0051] Figure 14 is a comparison of the liver tissue fibrosis indicator protein levels of mice in the CCl4-induced chronic mouse liver fibrosis injury model according to Example 3 of the present invention, a solvent control group, a CCl4 injection modeling group, and a CCl4 injection modeling group simultaneously given CCS1477 intraperitoneal injection treatment group.

[0052] Figure 15 is a comparison of the survival of mice in the model of acute-on-chronic liver failure induced by CCl4 and Klebsiella pneumonia (KP) in Example 4 of the present invention and the group treated with a single intraperitoneal injection of CCS1477, GNE207, and NEO2734 2 hours before KP injection.

[0053] Figure 16 is a comparison of laser Doppler blood flow imaging of the liver and kidneys of mice in the model of acute-on-chronic liver failure induced by CCl4 and Klebsiella pneumonia (KP) according to Example 4 of the present invention and the treatment group given a single intraperitoneal injection of CCS1477 2 hours before KP injection.

[0054] Figure 17 is a quantitative comparison of laser Doppler blood flow imaging of the liver and kidneys of mice in the model of acute-on-chronic liver failure induced by CCl4 and Klebsiella pneumonia (KP) according to Example 4 of the present invention and the treatment group given a single intraperitoneal injection of CCS1477 2 hours before KP injection. DETAILED DESCRIPTION

[0055] the term

[0056] As used herein, the term "prevent" or "prevent" refers to a reduction in the risk of acquiring a disease or disorder (i.e., halting the development of at least one clinical symptom of a disease in a subject who may be at risk or predisposed to the disease but who has not yet experienced or displayed symptoms of the disease).

[0057] In this application, the term "treatment" refers to any form of treatment that provides an effect to an individual who has a disease or is at risk of developing a disease, including improvement of the individual's condition (e.g., one or more symptoms), delay in progression of the disease, delay in onset of symptoms, or blunting of symptom progression. Therefore, the terms "treatment" and "prevention" do not refer to a cure or complete elimination of symptoms.

[0058] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0059] The embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and are not to be construed as limiting the present invention.

[0060] In the following examples, the experimental methods without specific conditions are carried out according to conventional methods and conditions, or according to the product specifications. The reagents and raw materials used in the present invention are all commercially available.

[0061] Example 1

[0062] This example studies the therapeutic effects of different p300 / CBP inhibitors (A-485, CBP30, and CCS1477) administered at different concentrations on cellular inflammatory factors after LPS stimulation.

[0063] Experimental Procedure: RAW264.7 macrophages were seeded in 6-well plates and incubated with conditioned medium for 24 hours. A control group, an LPS model control group, and treatment groups were designated. Treatment groups were treated with various p300 / CBP inhibitors at varying concentrations. The control group received the same dose of dimethyl sulfoxide as the drug-treated group. Both the LPS model control and treatment groups were stimulated with 500 ng / mL LPS for 6 hours. Treatment groups were administered 2 hours before LPS stimulation and then co-stimulated with LPS for 6 hours. Following LPS stimulation, cellular RNA was extracted using the Trizol method and subjected to real-time quantitative PCR to evaluate mRNA expression levels of the inflammatory factors TNF-α, IL-1β, IFNβ, and NOS2. Alternatively, cell supernatants were collected and analyzed by enzyme-linked immunosorbent assay to evaluate protein expression levels of the inflammatory factors TNF-α, IL-1β, and IL-6.

[0064] Results: Figure 1 shows the mRNA expression levels of inflammatory factors in the drug treatment group, blank control group, and model control group after 6 hours of LPS stimulation. Figure 2 shows the protein expression levels of inflammatory factors in the drug treatment group, blank control group, and model control group after 6 hours of LPS stimulation.

[0065] Conclusion: A-485, CBP30, and CCS1477 in the treatment group can all alleviate the increase in inflammatory cytokine expression levels induced by LPS stimulation, and the therapeutic effect is concentration-dependent. Among them, A-485 and CBP30 have similar efficacy, and both are weaker than CCS1477.

[0066] Example 2

[0067] This example studies the therapeutic effect of the p300 / CBP inhibitor CCS1477 on LPS-induced acute liver failure in mice.

[0068] Experimental Procedure: The groups were divided into a model control group, a drug treatment group, a drug toxicity test group, and a blank control group. Ten-week-old, specific pathogen-free C57 male mice (to avoid the influence of estrogen) were used as experimental animals. The modeling protocol consisted of a single intraperitoneal injection of 2 mg / kg LPS (lipopolysaccharide) combined with 250 mg / kg D-GalN (D-galactosamine) dissolved in PBS. Administration: A single intraperitoneal injection was administered 4 hours before LPS injection at drug concentrations of 20 mg / kg and 40 mg / kg, with a drug solution injection dose of 200-300 μL per mouse (the drug solution dose and solvent ratio were adjusted based on the average weight of the mice; multiple experiments confirmed that the solvent ratio did not affect the experimental results). The drug solvent was 10% DMSO (dimethyl sulfoxide), 25-40% PEG300 (polyethylene glycol 300), and 65-50% PBS (phosphate buffered saline). The drug administration and LPS injections were performed as quickly as possible, with the time difference between the first and last injections in mice being 5-10 minutes (the shorter the difference, the better). The model control group received the drug vehicle 4 hours before LPS injection; the drug treatment group received the drug solution 4 hours before LPS injection; the drug toxicity test group received the drug solution 4 hours before PBS injection; and the blank control group received the drug vehicle 4 hours before PBS injection. Two parallel groups were set up: an observation group and a sampling group. One group was used to observe 24-hour survival, and the other was used for sampling. The 24-hour survival of the observation group mice was observed: the number of deaths in each group was recorded starting 4 hours after LPS injection and every hour thereafter. Mice in the sampling group were anesthetized 4 hours after LPS injection, their eyes enucleated, and blood was collected before being sacrificed. Liver removal was then performed. Evaluation criteria included 24-hour survival rate, gross appearance of the liver, liver function assessment using serum ALT and AST, and pathological sections including HE staining, TUNEL apoptosis staining, and transcriptional levels of inflammatory factors in liver tissue.

[0069] Results: Figure 3 shows the 24-hour survival rates of the model control group, drug treatment group (CCS1477, doses of 20 mg / kg and 40 mg / kg, respectively), drug toxicity detection group (CCS1477, dose of 40 mg / kg), and blank control group. Figure 4 shows the gross appearance of the livers of the model control group, drug treatment group (CCS1477, 40 mg / kg), and blank control group. Figure 5 shows HE staining of liver tissues of the model control group, drug treatment group (CCS1477, 40 mg / kg), and blank control group. Figure 6 shows TUNEL apoptosis staining of liver tissues of the model control group, drug treatment group (CCS1477, 40 mg / kg), and blank control group. Figure 7 shows the liver function evaluation indicators: serum ALT and AST levels of the model control group, drug treatment group (CCS1477, doses of 20 mg / kg and 40 mg / kg, respectively), drug toxicity detection group (CCS1477, dose of 40 mg / kg), and blank control group. Figure 8 shows the transcriptional levels of inflammatory factors TNFα, IL-1β, IL-6, and NOS2 in the liver tissues of the model control group, the drug treatment group (CCS1477, at doses of 20 mg / kg and 40 mg / kg, respectively), and the blank control group.

[0070] In the LPS-induced acute liver failure model, the 24-hour survival rates of mice in each group are shown in Figure 3: The 24-hour survival rates of mice in the model control group vs. the drug-treated group (CCS1477, 40 mg / kg) vs. the drug-treated group (CCS1477, 20 mg / kg) were 25% vs. 83.33% vs. 66.67%, p<0.0001. The 24-hour survival rates of mice in the drug toxicity group and the blank control group were both 100%. Gross appearance of the livers of mice in each group is shown in Figure 4: The drug-treated group showed significant improvement in liver morphology. Compared with the blank control group, the livers of mice in the model control group were significantly darker and larger in size, while these abnormalities in gross liver morphology were significantly alleviated in the drug-treated group. HE staining and TUNEL apoptosis staining of mice in each group are shown in Figures 5 and 6: HE staining of liver tissue in the model control group revealed severe histological abnormalities, including destruction of liver tissue architecture, hepatocyte necrosis, congestion, and inflammatory cell infiltration. These effects were significantly improved in the drug-treated groups; similarly, TUNEL apoptosis staining showed increased hepatocyte apoptosis in the liver tissue of model control mice, while treatment with the drug CCS1477 significantly reduced apoptotic cells in the liver. The transcriptional levels of inflammatory factors in the liver tissue of mice in each group are shown in Figure 8: Compared with the blank control group, the expression of pro-inflammatory mediators (IL-1β, IL-6, and TNF-α) in the model control group was significantly increased. In the drug-treated groups, CCS1477 treatment significantly reduced the levels of inflammatory cytokines in liver tissue, which is consistent with the in vitro results (Figures 1 and 2).

[0071] Conclusion: Figure 3 shows that after LPS-induced acute liver failure, the survival rate of mice treated with CCS1477 was significantly increased. Figure 8 shows that after LPS-induced acute liver failure, the transcriptional levels of inflammatory factors in liver tissue were significantly elevated. However, after CCS1477 treatment, the elevated inflammatory factors were significantly reduced at the transcriptional level, alleviating inflammation. Figures 4, 5, 6, and 7 show that after LPS-induced acute liver failure, liver tissue was severely damaged. However, after CCS1477 treatment, liver tissue damage was effectively restored, and hepatocyte apoptosis was reduced.

[0072] In summary, it was shown that treatment with the p300 / CBP inhibitor CCS1477 had a therapeutic effect on liver damage in mice in the LPS-induced acute liver failure model.

[0073] Example 3

[0074] This example studies the therapeutic effect of p300 / CBP inhibitors on CCl4-induced chronic liver fibrosis in mice.

[0075] Experimental procedures: The groups were divided into a model control group, a drug treatment group, and a blank control group. Six- to eight-week-old, specific pathogen-free male C57 mice (to avoid the effects of estrogen) were used as experimental animals. The model control group received intraperitoneal injections of 10% CCl₄ solution diluted in olive oil twice weekly for 8 weeks at a dose of 1 mL / kg. The drug treatment group received concurrent drug treatment starting the week of CCl₄ injection. The drug treatment regimen consisted of 5 mg / kg / day of CCS1477, administered intraperitoneally (100 μl / mouse) in a solution of 10% DMSO, 15% Peg300, and 75% PBS. The blank control group received intraperitoneal injections of the corresponding dose of olive oil twice weekly for 8 weeks. Blood samples were collected 24 hours after the last CCl₄ injection and 4 hours after drug administration (with a 24-hour fast before sampling). Liver samples were obtained after anesthesia and sacrifice. Evaluation indicators: gross appearance of the liver, liver function evaluation: serum ALT and AST, pathological sections: Sirius red staining, immunohistochemical staining (α-SMA and COL1A1) and transcription and protein levels of liver tissue fibrosis factors, etc.

[0076] Results: Figure 9 shows the gross appearance of the livers in the model control group, the drug treatment group (CCS1477, 5 mg / kg / day), and the blank control group. Figure 10 shows Sirius red staining of the liver tissues in the model control group, the drug treatment group (CCS1477, 5 mg / kg / day), and the blank control group. Figure 11 shows immunohistochemical staining (α-SMA and COL1A1) in the model control group, the drug treatment group (CCS1477, 5 mg / kg / day), and the blank control group. Figure 12 shows serum ALT and AST levels, indicators of liver function evaluation, in the model control group, the drug treatment group (CCS1477, 5 mg / kg / day), and the blank control group. Figures 13 and 14 show the expression of fibrosis factors (α-SMA and COL1A1) at the transcriptional and protein levels in the model control group, the drug treatment group (CCS1477, 5 mg / kg / day), and the blank control group, respectively.

[0077] In the CCl4-induced chronic liver fibrosis injury model, the gross appearance of the livers of mice in each group is shown in Figure 9. The drug-treated group showed significant improvement in liver morphology. Compared with the blank control group, the livers of mice in the model control group showed increased surface granularity, condensed livers, and a firm texture. However, the abnormal gross morphology of the livers in the drug-treated groups was significantly alleviated. Sirius red staining of the livers of mice in each group is shown in Figure 10. Sirius red staining of the liver tissues of mice in the model control group revealed increased fibrillary and collagen deposition, indicating increased liver fibrosis. However, collagen deposition and fibrosis were significantly reduced in the liver tissues of mice in the CCS1477-treated group. Immunohistochemical staining of liver tissues (α-SMA and COL1A1) for each group is shown in Figure 11. The α-SMA and COL1A1-positive areas were significantly increased in the model control group but decreased in the drug-treated groups. Figures 13 and 14 show the expression of fibrosis factors (α-SMA and COL1A1) at the transcriptional and protein levels in each group of mice. Consistent with Figure 11, the expression of α-SMA and COL1A1 at the transcriptional and protein levels was significantly increased in the model control group, but decreased in the drug treatment group. Figure 12 shows the serum liver function evaluation indicators ALT and AST in each group of mice. ALT and AST were significantly elevated in the model control group, but were alleviated and decreased in the drug treatment group.

[0078] Conclusion: As shown in Figure 9, after CCl4-induced chronic liver fibrosis injury, the liver tissue showed obvious granular condensation and hardening, and CCS1477 treatment can improve the abnormal morphology of chronic liver injury. As shown in Figures 10, 11, 13, and 14, after CCl4-induced chronic liver fibrosis injury, the expression of fibrosis factors increased significantly, and fibrous cords in the liver tissue were clearly formed. CCS1477 treatment can improve the expression of fibrosis factors in the liver tissue and reduce the formation of fibrous cords in the liver tissue. As shown in Figure 12, after CCl4-induced chronic liver fibrosis injury, the liver tissue was severely damaged. After CCS1477 treatment, the liver tissue damage was effectively restored, and the levels of liver enzymes ALT and AST were reduced.

[0079] In summary, it was shown that treatment with the p300 / CBP inhibitor CCS1477 had a therapeutic effect on liver damage in mice induced by CCl4-induced chronic liver fibrosis.

[0080] Example 4

[0081] This example studies the therapeutic effects of different p300 / CBP inhibitors on CCl4 and Klebsiella pneumoniae (KP)-induced acute-on-chronic liver failure in mice.

[0082] Experimental procedures: The groups were set up as model control group and drug treatment group. Experimental animals were 6-8 weeks old, specific pathogen-free C57 male mice (to avoid the influence of estrogen). The modeling scheme was divided into chronic modeling stage, acute challenge stage and KP infection stage. In the chronic modeling stage, mice were intraperitoneally injected with 10% CCl4 solution diluted in olive oil. The injection time was 8-12 weeks, the injection frequency was twice a week, and the injection dose was 1mL / kg. In the acute challenge stage, 72 hours after the chronic modeling stage, a single intraperitoneal injection of 10% CCl4 solution diluted in olive oil was performed at a dose of 2mL / kg. In the KP infection stage, 24 hours after the acute challenge stage, a single intraperitoneal injection of KP bacterial solution was performed at a concentration of 5000 CFU / mL, and the dose was 100uL / mouse. The model control group was established according to the above protocol and received a single intraperitoneal injection of the drug vehicle 2 hours before KP injection. The drug treatment group received a single intraperitoneal injection of the drug solution at a dose of 10 mg / kg 2 hours before KP injection. The drug solution consisted of 10% DMSO, 25% PEG-300, and 65% PBS. Two parallel groups were set up: an observation group and a blood flow monitoring group. One group was used to monitor 72-hour survival, while the other group was used to monitor liver and kidney blood flow. 72-hour survival was monitored in the observation group. The number of deaths in each group was recorded starting 24 hours after the acute CCl4 injection. Deaths were recorded every hour from 24 to 48 hours, and every 2 hours from 48 to 72 hours. Half of the mice in the blood flow monitoring group were anesthetized and opened 48 hours after the acute CCl4 injection, and the other half were anesthetized 60 hours after the acute CCl4 injection. Liver and kidney blood flow were monitored using Doppler flowmetry, and livers were subsequently harvested. Evaluation indicators: 72-hour survival rate, liver and kidney blood flow, etc.

[0083] Results: Figure 15 shows the 7-day survival rates of mice in the model control group and the drug-treated groups (CCS1477, 10 mg / kg), (NEO2734, 5 mg / kg), and (GNE207, 10 mg / kg). Figures 16 and 17 show the liver and kidney blood flow of mice in the model control group and the drug-treated group (CCS1477, 10 mg / kg) under laser Doppler monitoring.

[0084] In the CCl4 and Klebsiella pneumoniae (KP)-induced acute-on-chronic liver failure experiment in mice, the 72-hour survival rates of mice in each group are shown in Figure 15 : the 72-hour survival rates of mice in the model control group and the drug-treated group were (0.00% vs. 50.00%, p<0.0001). Liver and kidney blood flow patterns in each group at 48 and 60 hours, as monitored by Doppler flowmetry, are shown in Figures 16 and 17 .

[0085] Conclusion: As shown in Figure 15, after mice induced with acute-on-chronic liver failure (ACHF) using CCl4 and Klebsiella pneumoniae (KP), the survival rates of mice in the drug treatment groups treated with CCS1477, NEO2734, and GNE207 were all increased, with CCS1477 showing the greatest efficacy. As shown in Figures 16 and 17, after mice induced with ACHF using CCl4 and Klebsiella pneumoniae (KP), the model control group experienced significant liver and kidney blood flow impairment, while the CCS1477-treated group showed significant improvement in liver and kidney blood flow impairment.

[0086] In summary, it was shown that the p300 / CBP inhibitor CCS1477 had a therapeutic effect on liver damage in mice with acute-on-chronic liver failure induced by CCl4 and Klebsiella pneumoniae (KP).

[0087] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A method for preventing and / or treating liver damage diseases by targeted inhibition of p300 / CBP in a subject in need thereof.

2. The method according to claim 1, in, The targeted inhibition of p300 / CBP includes targeting the HAT domain and the BRD domain of p300 / CBP.

3. The method according to claim 1, in, The treatment or prevention includes producing one or more of the following effects on the subject: inhibiting the secretion of inflammatory factors of immune cells, improving liver tissue damage, reducing liver cell apoptosis, lowering liver enzyme levels, improving liver and kidney blood flow disorders, and improving survival rate.

4. The method according to claim 1, in, The targeted inhibition of p300 / CBP comprises administering to the subject a therapeutically effective amount of a p300 / CBP inhibitor, which includes CCS1477, GNE207, NEO2734, A-485 and CBP30.

5. The method according to claim 1, in, The liver injury diseases include acute liver injury, chronic liver injury and / or liver failure and acute-on-chronic liver failure.

6. The method according to claim 5, in, The liver injury disease is acute liver injury, and the dosage of the p300 / CBP inhibitor administered to the subject is 20-40 mg / Kg for a single administration, for example, the dosage of the drug administered to the subject is 20 mg / Kg, 25 mg / Kg, 30 mg / Kg, 35 mg / Kg and 40 mg / Kg for a single administration.

7. The method according to claim 5, in, The liver damage disease is chronic liver damage, and the p300 / CBP inhibitor is administered to the subject at a dosage of 3-10 mg / Kg·day, for example, 5 mg / Kg·day.

8. The method according to claim 5, in, The liver damage disease is acute-on-chronic liver failure, and the dosage of the p300 / CBP inhibitor administered to the subject is 5-20 mg / Kg for a single administration, for example, the dosage of the drug administered to the subject is 5 mg / Kg, 10 mg / Kg, 15 mg / Kg and 20 mg / Kg for a single administration.

9. Use of a p300 / CBP inhibitor in the preparation of a medicament for preventing and / or treating liver damage diseases.

10. The use according to claim 9, in, The p300 / CBP inhibitor targets the HAT domain and the BRD domain of p300 / CBP.

11. The use according to claim 9, in, The liver injury diseases include acute liver injury, chronic liver injury and / or liver failure and acute-on-chronic liver failure.

12. The use according to claim 9, in, The p300 / CBP inhibitors include CCS1477, GNE207, NEO2734, A-485 and CBP30.

13. A drug for preventing and / or treating liver damage diseases, wherein the active ingredient of the drug comprises a p300 / CBP inhibitor.

14. The drug according to claim 13, in, The p300 / CBP inhibitors include compounds targeting the HAT domain and the BRD domain of p300 / CBP.

15. The drug according to claim 13, in, The p300 / CBP inhibitors include CCS1477, GNE207, NEO2734, A-485 and CBP30.

16. The drug according to claim 13, in, The liver injury diseases include acute liver injury, chronic liver injury and / or liver failure and acute-on-chronic liver failure.

17. The drug according to claim 13, in, The dosage forms of the drug include injection, tablet, capsule, oral granule and enema.

18. A pharmaceutical composition for preventing and / or treating liver damage diseases, comprising the drug according to claim 13 and a pharmaceutically acceptable excipient, wherein the excipient is one selected from a binder, a filler, a coating polymer, a plasticizer, a glidant, a disintegrant and a lubricant, or any combination thereof.

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