阅读说明：本技术 一种治疗肝纤维化、硬皮病的化合物及其应用 (Compound for treating hepatic fibrosis and scleroderma and application thereof ) 是由 卢伟强 章涵堃 陈思 姜兴武 蒋蓓尔 刘明耀 于 2019-09-18 设计创作，主要内容包括：本发明属于医药领域,具体公开了一种治疗纤维化相关疾病的化合物LQ-2-191。本发明提供了以大麻素受体CB2为作用靶点在抗纤维化药物开发中的应用。所述的LQ-2-191可在体内外特异性地激活CB2,通过作用于CB2抑制纤维化部位的炎症反应并有效缓解纤维化的症状。本发明通过动物实验证实,LQ-2-191能够有效减轻胆管结扎诱导的大鼠肝纤维化和博来霉素诱导的小鼠硬皮病模型纤维化程度。所述抗纤维化药物以大麻素受体CB2为作用靶点,可以明显抑制肝纤维化、硬皮病的发生和发展,可用于制备抗肝纤维化、防治肝硬化和硬皮病的药物。(The invention belongs to the field of medicines, and particularly discloses a compound LQ-2-191 for treating fibrosis related diseases. The invention provides application of a target point with cannabinoid receptor CB2 as an action in developing anti-fibrosis drugs. The LQ-2-191 can specifically activate CB2 in vivo and in vitro, inhibit inflammatory reaction at a fibrosis part by acting on CB2 and effectively relieve the symptoms of fibrosis. Animal experiments prove that the LQ-2-191 can effectively relieve rat hepatic fibrosis induced by bile duct ligation and mouse scleroderma model fibrosis induced by bleomycin. The anti-fibrosis drug takes cannabinoid receptor CB2 as an action target, can obviously inhibit the occurrence and the development of hepatic fibrosis and scleroderma, and can be used for preparing drugs for resisting hepatic fibrosis and preventing and treating liver cirrhosis and scleroderma.)

1. The application of the compound LQ-2-191 in preparing the medicament for treating fibrosis related diseases is characterized in that the molecular formula of the LQ-2-191 is as follows: c₁₈H₂₃F₃N₄O₂The structure is shown as formula (1):

2. the use of claim 1, wherein LQ-2-191 reduces liver, skin inflammation by inhibiting inflammatory mediator production or infiltration; alternatively, said LQ-2-191 ameliorates fibrotic symptoms by reducing collagen deposition.

3. The use of claim 2, wherein the inflammatory mediator is TNF- α, TGF- β.

4. The use of claim 1, wherein the fibrosis-related disease drug targets cannabinoid receptor CB 2.

5. The use according to claim 1, wherein LQ-2-191 is administered in an amount of 1-20mg per kilogram body weight of the individual.

6. The pharmaceutical composition is characterized by comprising an active ingredient LQ-2-191 and a pharmaceutically acceptable carrier, wherein the structure of the LQ-2-191 is shown as a formula (1):

use of LQ-2-191 or a composition according to claim 6 for the preparation of a medicament for the inhibition and/or treatment of inflammatory mediators produced during scleroderma, liver fibrosis or both, wherein LQ-2-191 has the structure according to formula (1):

use of LQ-2-191 or a composition according to claim 6 for the manufacture of a medicament for reducing and/or treating collagen deposition during scleroderma, liver fibrosis or both, wherein LQ-2-191 has the structure according to formula (1):

9. the use of claim 7, wherein the LQ-2-191 is for reducing the stiffness and thickness of skin.

10. The use of claim 8, wherein LQ-2-191 is used to reduce the expression of ALT, AST and total bilirubin in the serum.

11. The use of claim 7, wherein the medicament is for the treatment of scleroderma.

12. The use according to claim 7, wherein the medicament is for the treatment of liver disease, which is a disease associated with liver fibrosis.

13. The use according to claim 12, wherein the liver disease is chronic hepatitis, nonalcoholic fatty liver, cirrhosis or liver cancer and a disease caused by liver damage.

Technical Field

The invention belongs to the field of medicines, and particularly relates to a compound for treating hepatic fibrosis and scleroderma and application thereof.

Background

Fibrosis, a pathological condition caused by excessive tissue repair, generally occurs during wound healing after tissue injury. Examples of conditions associated with tissue fibrosis are numerous and generally include the formation of skin scarring, liver fibrosis, kidney fibrosis, pulmonary interstitial fibrosis, glomerulonephritis, heart failure (ischemic and non-ischemic), scleroderma, and the like.

Scleroderma, systemic sclerosis, is an autoimmune disease, the most prominent external features being skin thickening and arteriolar damage due to collagen accumulation. Two forms of sclerosis are currently common in the clinic: local sclerosis and systemic sclerosis. Topical sclerosis affects mainly the skin of the face, hands and feet. Systemic sclerosis may also develop in internal organs including the kidney, heart, lung and gastrointestinal tract, in addition to the skin. The current data indicate that the 10-year survival rate for patients with localized systemic sclerosis is 75%; nearly 10% of the years have progressed to pulmonary hypertension in 10 to 20 years. The 10-year survival rate for systemic sclerosis patients is only 55%, the most common cause of death being failure of the lungs, heart and kidneys. In addition, these patients are at a slightly increased risk of developing cancer.

Hepatic fibrosis is one of the key processes of the progression of various acute and chronic liver diseases such as liver cirrhosis and is also the most important risk factor of liver cancer. Liver cancer is the cancer with the highest mortality in the world, the pathogenesis process of the liver cancer is generally acute and chronic liver injury caused by alcohol, virus and other pathogenic factors, the liver is excessively repaired under the conditions of continuous injury and strong inflammation reaction, the balance of collagen fiber synthesis and degradation is broken, a large amount of extracellular matrix protein is excessively accumulated in the liver, liver fibrosis and liver cirrhosis are caused, and the liver cancer is possibly developed finally, and the pathogenesis mode is the main pathogenesis mode of liver cancer in China. Liver fibrosis is a necessary way for the development of various chronic liver diseases such as hepatitis, non-alcoholic fatty liver disease and the like to cirrhosis, so that early prevention of liver fibrosis can slow down the progress of various liver diseases. A number of animal experiments and clinical trials have demonstrated that liver fibrosis is a reversible process. Improving liver fibrosis can not only prevent inflammatory necrosis of liver cells and reduce abnormal liver metabolism, but also slow down the progress of cirrhosis and liver cancer, thereby effectively increasing survival time of patients and improving the life quality of patients.

Cannabinoid receptors and their downstream pathways are intimately involved in fibrosis caused by a variety of inflammations. In the liver, after the CB1 receptor in the fibroblast is activated, the hepatic stellate cell can be activated, the division speed of the myofibroblast is accelerated, and the hepatic fibrosis is promoted. After activation of the CB2 receptor expressed in the Kunpan cells, the anti-inflammatory factor for promoting the apoptosis of hepatic stellate cells can be secreted, and myofibroblast apoptosis can be induced, so that the effect of inhibiting hepatic fibrosis is achieved. Therefore, inhibition of the CB1 receptor or activation of the CB2 receptor may be effective in treating liver fibrosis. Also, for skin fibrosis caused by inflammation, inflammation can be effectively inhibited by activating the CB2 receptor, so that the occurrence and the development of skin fibrosis can be effectively inhibited. However, there is a great deal of clinical evidence that antagonists that inhibit the CB1 receptor can cause serious side effects such as depression, suicidal ideation tendencies, and the like. Therefore, the development of CB2 selective agonists is a promising new strategy for liver fibrosis treatment.

A number of cannabinoid receptor agonists are currently entering clinical research for anti-fibrosis. The Anabasum is a natural artificial cannabinoid analogue which enters the clinic and is used for treating the fibrotic diseases, and achieves more remarkable effects. However, the general selectivity of Anabasum for different cannabinoid receptor subtypes has limited its clinical utility. Therefore, there is a need to develop novel highly active, highly selective CB2 receptor agonists for use in the treatment of fibrotic diseases. Among them, CB2 receptor activation can inhibit or even reverse the occurrence of fibrosis.

Disclosure of Invention

Through large-scale drug screening, the invention discovers that LQ-2-191 can efficiently activate a G protein channel downstream of CB2, thereby effectively improving hepatic fibrosis and scleroderma. The LQ-2-191 inhibits inflammatory reaction of a fibrosis part by activating CB2 and effectively relieves the symptoms of fibrosis.

The technical problem to be solved by the invention is to provide a compound for treating hepatic fibrosis and scleroderma.

The invention provides application of a compound LQ-2-191 in preparation of a medicament related to treatment of a fibrotic disease.

Wherein, the fibrosis related disease drug takes cannabinoid receptor CB2 as an action target.

The drug is an anti-fibrosis drug named as LQ-2-191, and the molecular formula of the LQ-2-191 is as follows: c₁₈H₂₃F₃N₄O₂The structure is shown as formula (1):

in the invention, the LQ-2-191 can reduce inflammation of liver and skin by inhibiting the generation or infiltration of inflammatory mediators.

Wherein the inflammatory mediator is TNF-alpha, TGF-beta.

In the invention, the LQ-2-191 improves the fibrosis symptoms by reducing collagen deposition.

In the invention, the LQ-2-191 is administered in a dose of 1-20mg per kg of body weight of the subject.

The invention also provides a pharmaceutical composition, which comprises an effective amount of a compound shown as the formula (1) and a pharmaceutically acceptable carrier:

in the invention, the dosage form of the pharmaceutical composition is oral liquid.

The invention also provides application of the pharmaceutical composition in preparation of medicines for resisting hepatic fibrosis and scleroderma.

In the invention, the LQ-2-191 is injected into an animal individual (such as gavage), and the final concentration of the LQ-2-191 is 1-20 mg/kg. Preferably, the final concentration of the LQ-2-191 is 20 mg/kg.

The anti-fibrosis drug of the present invention can provide different effects when administered (administered) therapeutically. Generally, anti-fibrotic drugs can be formulated in non-toxic, inert, and pharmaceutically acceptable aqueous carrier media to form pharmaceutical compositions. Wherein the aqueous carrier medium is normal saline, phosphate buffer, dextrose solution, distilled water, glycerol, ethanol, and combinations thereof.

Wherein the pharmaceutical composition typically has a pH of about 5 to about 8; preferably, the pH is about 6-8, which may vary depending on the nature of the substance being formulated and the condition being treated.

LQ-2-191 of the present invention can be prepared in the form of an oral preparation, for example, by a conventional method using physiological saline or an aqueous solution containing glucose and other adjuvants. The pharmaceutical compositions are preferably manufactured under sterile conditions. The amount of active ingredient administered is a therapeutically effective amount, for example, about 1-20mg/kg per day. The specific dosage should also take into account such factors as the route of administration, the health of the patient, etc. Furthermore, LQ-2-191 of the present invention may also be used with other therapeutic agents.

The invention carries out weight monitoring on an induced fibrosis animal model, and detects the content of collagen I (colla I), alpha smooth muscle actin (alpha-SMA) and hydroxyproline in tissues, and the experimental result shows that the LQ-2-191 can reduce infiltration of inflammatory cells at a fibrosis part and deposition of collagen, thereby improving fibrosis symptoms.

The invention also provides application of LQ-2-191 or the pharmaceutical composition in preparing a medicament for inhibiting and/or treating inflammatory mediators generated in the processes of hepatic fibrosis and scleroderma.

Wherein the inflammatory mediator is TNF-alpha, TGF-beta.

Wherein, the medicine is used for treating liver diseases, and the liver diseases refer to diseases related to liver fibrosis; the liver disease is chronic hepatitis, non-alcoholic fatty liver, liver cirrhosis or liver cancer and diseases caused by liver injury.

Wherein the medicament is for reducing the stiffness and thickness of the skin.

Wherein the medicament is for use in treating scleroderma; the scleroderma is an autoimmune disease, which is mainly characterized by skin thickening and arteriolar lesions caused by collagen accumulation and fibrotic degeneration of heart, liver and lung organs.

The invention also provides application of the LQ-2-191 or the pharmaceutical composition in preparing a medicament for reducing and/or treating collagen deposited in the liver fibrosis and scleroderma processes.

Wherein the LQ-2-191 or the pharmaceutical composition is used for reducing the expression level of ALT, AST and total bilirubin in serum.

The beneficial effects of the invention include: the invention provides application of LQ-2-191 or a pharmaceutical composition in preparation of medicines for resisting hepatic fibrosis and scleroderma. The LQ-2-191 or the pharmaceutical composition can obviously inhibit the development of hepatic fibrosis and scleroderma. The invention develops the LQ-2-191 as a new anti-fibrosis drug or an auxiliary component thereof, has obvious anti-fibrosis effect and stable property, and provides a new way and means for treating hepatic fibrosis and scleroderma.

Drawings

FIG. 1 shows the results of the calcium flux test of example 1LQ-2-191 at the CB2 receptor; wherein, fig. 1a shows the calcium flow activity of a yang drug CP55940, and fig. 1b shows the calcium flow activity of LQ-2-191.

FIG. 2 shows that the activation of CB2 receptor by LQ-2-191 in example 1 is inhibited by the CB2 receptor antagonist AM 630.

FIG. 3 is a graph showing that example 1LQ-2-191 is capable of causing CB2 receptor desensitization; wherein, fig. 3a is calcium flux activity caused by DMSO, CP55940 and LQ-2-191 when stimulating CB2 twice, and fig. 3b is a statistical analysis chart of 3 a.

FIG. 4 shows the results of HTRF testing of example 1LQ-2-191 at the CB2 receptor; FIG. 4a shows HTRF test data of CP55940, and FIG. 4b shows HTRF test results of LQ-2-191.

FIG. 5 is a histopathological map of liver of the control group stained with H & E and animals administered LQ-2-191 in example 2.

FIG. 6 is a pathological view of liver tissues of animals treated with the control group stained with sirius red and administered with LQ-2-191 of example 2.

FIG. 7 shows the liver function markers including alanine Aminotransferase (ALT), aspartate Aminotransferase (AST) and total bilirubin (T-BIL) in the serum of the rat in example 2.

FIG. 8 is a graph of example 2LQ-2-191 reducing the amount of rat liver fibrosis related markers including TGF- β, Col1a1, α -SMA and TNF- α.

FIG. 9 is a graph showing that example 2LQ-2-191 reduces liver fibrosis in rat serum, including type III Procollagen (PCIII), type IV Collagen (CIV), Laminin (Laminin), and Hyaluronic Acid (HA).

FIG. 10 is a histopathological image of skin of the control group stained with H & E and the animals administered LQ-2-191 in example 3.

FIG. 11 is the skin histopathology of the control group stained with sirius red and the animals given LQ-2-191 of example 3.

FIG. 12 is a photograph of the skin histopathology of the control group and animals given LQ-2-191 in immunohistochemical fluorescent staining of example 3.

FIG. 13 is a graph of example 3LQ-2-191 reducing the amount of markers associated with mouse skin fibrosis, including TGF- β, Col1a1, α -SMA, and TNF- α.

Detailed Description

The present invention will be described in further detail with reference to the following specific examples, but the present invention is not limited to the following examples. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept, which is set forth in the following claims. The procedures, conditions, reagents, experimental methods and the like for carrying out the present invention are general knowledge and common general knowledge in the art except for the contents specifically mentioned below, and the present invention is not particularly limited.