HDAC6 and HSP90 bifunctional inhibitor and application method thereof

Abstract

The invention belongs to the technical field of medicine, and discloses an application method of an HDAC6 and HSP90 bifunctional inhibitor, a double-target drug can act on two different targets at the same time, so that a synergistic effect is achieved in the aspects of regulating a signal channel, regulating physiological functions and the like, and the treatment effect is improved. Due to the fact that the double-target medicine acts on the two targets at the same time, the drug resistance appearing in disease development is reduced, and the lasting time of the curative effect of the medicine is prolonged. By acting on the two targets at the same time, the double-target medicine can achieve the same curative effect under the lower dosage, and therefore the influence of the side effect on a patient is reduced. The double-target drug has the advantages of better curative effect, low drug resistance, small side effect, personalized treatment and the like. According to the present invention, two targets such as HSP90 and HDAC6 are combined, and an HDAC6 and HSP90 dual-function inhibitor is reported, and is used for treating inflammatory bowel diseases, such as ulcerative colitis and Crohn disease.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to an HDAC6 and HSP90 dual-function inhibitor and an application method thereof. Background Art

[0002] HSP90 (heat shock protein 90) is an important molecular chaperone protein and a member of the molecular chaperone protein family. HSP90 is involved in the folding, assembly and stabilization of many important proteins in cells and plays a key role under cell stress conditions. HSP90 helps these proteins achieve the correct conformation and function by binding to guest proteins.

[0003] HSP90 is widely present in cells, and its functions and regulatory mechanisms are extremely complex. HSP90 is mainly involved in regulating the conformational changes of proteins in many signal transduction pathways, thereby affecting the biological processes in cells. It plays an important role in cell proliferation, apoptosis, metabolism, and stress resistance. Since HSP90 is involved in the regulation of many cell signaling pathways, it is considered an important drug target. The abnormally high expression of HSP90 is closely related to the occurrence and development of many human diseases, such as malignant tumors, neurodegenerative diseases, autoimmune diseases, organ fibrosis, etc. For the indication of malignant tumors, there are many HSP90 inhibitors in the clinical trial stage. Currently, an HSP inhibitor has been marketed in Japan as a treatment for gastric stromal tumors.

[0004] In addition to the above diseases, the literature also reports that HSP90 is associated with the occurrence of ulcerative colitis. HSP90 inhibitors can improve the degree of pathological damage in the colon of DSS-induced mice, increase the weight and colon length of mice, and significantly reduce the disease activity index (DAI) score. Intraperitoneal injection of HSP90 inhibitors can reduce the expression and phosphorylation of MAPK pathway markers P38, JNK, and ERK, and reduce the content of AFT-2 and p53 downstream of the MAPK pathway. In addition, treatment with HSP90 inhibitors upregulated the expression of anti-apoptotic proteins BCL-2 and survivin in the colon of colitis mice, and downregulated the expression of apoptotic proteins caspase3 and BAX. After treatment with HSP90 inhibitors, it was also observed that the levels of inflammatory factors such as IL-6, MCP-1, IFN-γ, TNF, IL-12p70 decreased, and the level of IL-10 increased. The high expression of these inflammatory factors is closely related to ulcerative colitis.

[0005] In addition, abnormal activation of the NLRP3 inflammasome is associated with a variety of human inflammatory diseases, but no NLRP3 inhibitor drugs have been approved yet. In a study, researchers found that the component of the traditional herbal medicine licorice, Glycyrrhiza uralensis chalcone, effectively inhibited the activation of the NLRP3 inflammasome in vitro and in vivo. Further studies showed that Glycyrrhiza uralensis chalcone inhibited the NLRP3 inflammasome by binding to HSP90, inhibiting its ATPase activity, and disrupting the association between the co-chaperone SGT1 and HSP90-NLRP3. Importantly, in vivo experiments showed that administration of Glycyrrhiza uralensis chalcone significantly inhibited NLRP3 inflammasome activation and improved DSS-induced colitis in mice. Therefore, HSP90 can be used as a drug development target for the treatment of ulcerative colitis.

[0006] In addition, HDAC6 is a histone deacetylase, the full name of which is histone deacetylase 6. It is a member of the zinc-dependent deacetylase in the HDACs family, and is closely related to the regulation of organelle dynamics and protein degradation in the cytoplasm. HDAC6 is mainly expressed in the cytoplasm in human cells, and has dual enzyme activity, which can deacetylate non-histone proteins and histones, which is different from other HDACs. HDAC6 has diverse functions, including participating in biological processes such as protein degradation, cell movement, cell stress response, and apoptosis in cells. HDAC6 participates in the regulation of various important biological processes in cells by regulating microtubule structure, driving protein degradation pathways, affecting cell movement, and regulating autophagy.

[0007] Studies have found that HDAC6 plays an important role in the occurrence and development of many diseases, including cancer, neurodegenerative diseases, inflammatory diseases, etc. Therefore, HDAC6 has become one of the targets that has attracted much attention in the field of drug development. Studies have shown that HDAC6 is also associated with the onset of ulcerative colitis.

[0008] Treatment of mice with DSS-induced colitis with the HDAC6 inhibitor BML-281 can reduce colitis, weight loss, and disease pathology, including changes in colon structure and function. BML-281 treatment can induce broad-spectrum anti-inflammatory effects and prevent the infiltration and activation of key immune cells in the intestinal epithelial lamina propria. Among different immune cells, BML-281 specifically inhibits the infiltration of CD19+B cells into the inflamed colonic lamina propria. This study supports the use of HDAC6 as an anti-inflammatory strategy for treating colonic inflammation that progresses to IBD. Also in another work, the researchers found that the HDAC6 inhibitor WT161 can play a protective role in a colitis model by blocking NLRP3 inflammasome activation, disrupting ASC spot formation, and reducing NLRP3 expression. In summary, HDAC6 is also a drug development target for the treatment of ulcerative colitis.

[0009] Through the above analysis, the problems and defects of the prior art are as follows:

[0010] Existing drugs, nintedanib and pirfenidone, have poor therapeutic effects on inflammatory bowel disease, have toxic side effects, and are prone to drug resistance. Summary of the invention

[0011] In view of the problems existing in the prior art, the present invention provides an application method of a dual-function inhibitor of HDAC6 and HSP90.

[0012] The present invention is achieved in that a method for applying a dual-function inhibitor of HDAC6 and HSP90 comprises:

[0013] S101, the therapeutic application of HDAC6 / HSP90 dual-function inhibitors in inflammatory bowel diseases, including ulcerative colitis and Crohn's disease.

[0014] Furthermore, the molecular structure of the HDAC6 / HSP90 dual-function inhibitor is as follows:

[0015]

[0016] R is one of the following molecular formulas:

[0017]

[0018] Further, the HDAC6 and HSP90 dual-function inhibitor represents the structural formula of compound 1:

[0019]

[0020] In combination with the above technical solutions and the technical problems solved, the advantages and positive effects of the technical solutions to be protected by the present invention are as follows:

[0021] First, the research and development of dual-target and multi-target drugs has attracted much attention in recent years, mainly because they have shown significant advantages in terms of therapeutic effects and patient experience. First of all, enhancing the efficacy is one of the main advantages of dual-target drugs. By acting on two different targets at the same time, dual-target drugs can exert a synergistic effect in regulating signal pathways and regulating physiological functions, thereby significantly improving the therapeutic effect. Secondly, reducing drug resistance is also an important advantage of dual-target drugs. Since dual-target drugs intervene in two targets at the same time, the likelihood of drug resistance during the development of the disease is reduced and the duration of the drug's efficacy is prolonged. In addition, reducing side effects is also one of its significant advantages. Dual-target drugs can achieve the same therapeutic effect at a lower dose, thereby reducing the side effects of drugs on patients and improving patients' tolerance and quality of life. Finally, improving personalized treatment capabilities gives dual-target drugs a unique advantage in modern medicine. By selecting different target combinations according to the specific circumstances of the patient, dual-target drugs can achieve more personalized treatment plans and improve the pertinence and effectiveness of treatment.

[0022] Based on the above advantages, the present invention combines two targets, HSP90 and HDAC6, and reports a bifunctional inhibitor of HDAC6 and HSP90, which is specifically used to treat inflammatory bowel disease (IBD), including ulcerative colitis and Crohn's disease. The bifunctional inhibitor can regulate inflammatory responses and cellular stress responses at multiple levels by simultaneously inhibiting HDAC6 and HSP90, thereby effectively reducing intestinal inflammation and promoting the repair and regeneration of diseased tissues. Specifically, the inhibitory effect of HDAC6 helps to regulate the deacetylation state of intracellular proteins, reduce inflammatory responses, and promote cell survival and repair; while the inhibitory effect of HSP90 interferes with its function as a molecular chaperone, affects multiple signal transduction pathways, further reduces inflammatory responses and inhibits pathological processes. Through this dual mechanism of action, the bifunctional inhibitor can not only improve the therapeutic effect, but also reduce the occurrence of drug resistance, reduce the risk of side effects, and achieve a more personalized and efficient treatment plan.

[0023] In summary, the HDAC6 and HSP90 bifunctional inhibitors described in the present invention, with their significant advantages such as enhanced efficacy, reduced drug resistance, reduced side effects and improved personalized treatment, show broad application prospects and significant technological progress in the treatment of inflammatory bowel disease, are in line with the current hot trends in the field of drug research and development, and have important clinical application value and market potential.

[0024] The present invention provides a dual-function inhibitor that combines two targets, HSP90 and HDAC6, specifically for the treatment of inflammatory bowel disease (IBD), including ulcerative colitis and Crohn's disease. By inhibiting HDAC6 and HSP90 at the same time, the inhibitor can effectively regulate inflammatory response and cellular stress response, thereby reducing intestinal inflammation and promoting the repair and regeneration of diseased tissues.

[0025] Second, the HDAC6 and HSP90 dual-function inhibitor of the present invention has the following significant technical effects in the treatment of inflammatory bowel disease:

[0026] 1. Enhanced anti-inflammatory effect: By simultaneously inhibiting HDAC6 and HSP90, dual regulation of inflammatory response can significantly reduce intestinal inflammation and improve patient symptoms.

[0027] 2. Promote tissue repair: Regulate intracellular protein status and signal transduction pathways, promote the repair and regeneration of damaged intestinal tissue, and improve the speed and quality of tissue function recovery.

[0028] 3. Reduce the risk of side effects: Bifunctional inhibitors can achieve significant therapeutic effects at lower doses, reduce drug-related side effects, and improve patient tolerance and treatment compliance.

[0029] 4. Improve treatment efficiency: Optimize treatment plans through precise dosage and dosing frequency adjustments, accelerate disease remission and recovery process, and improve overall treatment efficiency.

[0030] 5. Personalized treatment: Continuously monitor the patient's inflammatory indicators and condition changes, adjust the treatment plan based on individual differences, achieve personalized and precise treatment, and meet the treatment needs of different patients.

[0031] In summary, the HDAC6 and HSP90 dual-function inhibitor and its application method provided by the present invention significantly improve the therapeutic effect of inflammatory bowel disease through an innovative dual-target inhibition mechanism, have broad clinical application prospects and significant technological progress, and meet the creativity requirements of the Patent Law.

[0032] Third, the expected benefits and commercial value after the technical solution of the present invention is transformed as follows: the HSP90 and HDAC6 bifunctional inhibitors involved in the present invention have shown excellent therapeutic effects on inflammatory bowel disease, with complete in vitro and in vivo data, and can be transformed into results. The estimated transformation amount is RMB 50 million.

[0033] The technical solution of the present invention fills the technical gap in the industry at home and abroad: At present, there is no HSP90 and HDAC6 dual-function inhibitor clinically used in the treatment of inflammatory bowel disease, including ulcerative colitis and Crohn's disease. The proposal and application of this technical solution can fill this gap, provide a basis for the dual-targeted treatment of inflammatory bowel disease, and provide a new and effective treatment for patients with inflammatory bowel disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 It is a flow chart of the application method of the HDAC6 and HSP90 dual-function inhibitor provided by an embodiment of the present invention.

[0035] Figure 2 It is a control experiment diagram provided by an embodiment of the present invention;

[0036] Figure 3 The therapeutic effects of compound 2 provided in the examples of the present invention at different doses on mice with DSS-induced ulcerative colitis;

[0037] Figure 4 The survival status of mice in a chronic toxicity experiment of compound 2 provided in an example of the present invention;

[0038] Figure 5 This is a schematic diagram of the disease activity index results of compound 2 provided in an example of the present invention at different doses on DSS-induced ulcerative colitis mice;

[0039] Figure 6 It is a schematic diagram of the body weight results of mice in a chronic toxicity experiment of compound 2 provided in an example of the present invention. DETAILED DESCRIPTION

[0040] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0041] like Figure 1 As shown, the application method of an HDAC6 and HSP90 dual-function inhibitor provided in an embodiment of the present invention comprises the following steps:

[0042] S101, the therapeutic application of HDAC6 / HSP90 dual-function inhibitors in inflammatory bowel diseases, including ulcerative colitis and Crohn's disease.

[0043] The molecular structure formula of the HDAC6 / HSP90 dual-function inhibitor provided in the embodiment of the present invention is:

[0044]

[0045] R is one of the following molecular formulas:

[0046]

[0047] The structural formula of the representative compound 2 and its in vivo anti-ulcerative colitis activity are shown below:

[0048]

[0049] Compound 2 can inhibit the activity of HSP90 and HDAC6 at the same time. The results are shown in the following table:

[0050]

[0051] As can be seen from the table above, compound 2 can inhibit HSP90 and HDAC6 at the same time at nM concentration, and is a dual-target inhibitor. In addition, the compound shows a lower inhibitory effect on hERG, indicating that it has no potential cardiotoxicity.

[0052] Figure 2 Compound 2 and the positive control drug 5-aminosalicylic acid (5-ASA) were both administered orally, and the dosage of compound 2 was (5 mg / kg, 10 mg / kg, 20 mg / kg), twice a day. Figure 3 It can be seen from the experimental results that compound 2 at three oral administration concentrations (5 mg / kg, 10 mg / kg, 20 mg / kg) can not only effectively slow down the weight loss of mice compared with the DSS modeling group, but also effectively prolong the length of the mouse colon. The therapeutic effect is slightly better than the positive control drug 5-aminosalicylic acid (5-ASA) at an oral dose of 200 mg / kg, proving that the compound has the potential to treat ulcerative colitis.

[0053] In terms of safety, the acute and chronic toxicity of compound 2 were evaluated. First, for acute toxicity, mice were orally administered 1000 mg / kg, 2000 mg / kg, and 3000 mg / kg of compound 2 at one time. There were 6 mice in each group, half male and half female. The survival status of the mice was recorded after 14 days. The results are shown in the following table:

[0054]

[0055] From the results, we can see that the LD 50 The value was greater than 3000 mg / kg, which was almost non-toxic. We then conducted a chronic toxicity experiment, orally administering 100 mg / kg of compound 2 to mice once a day, with a total of 6 mice, half male and half female. After 30 days, the survival status of the mice was recorded. The results are as follows Figure 6 As shown:

[0056] After 30 days of administration, no mice died, and the weight of the mice gradually increased, and the weight remained normal two days after drug withdrawal, which further proved that compound 2 had extremely low toxicity.

[0057] Example 1: Application of HDAC6 and HSP90 dual-function inhibitors in ulcerative colitis

[0058] 1. Experimental Background

[0059] Ulcerative colitis (UC) is a chronic inflammatory bowel disease that mainly affects the colon and rectal mucosa, and patients often present with symptoms such as abdominal pain, diarrhea, and blood in the stool. Traditional treatment methods include anti-inflammatory drugs, immunosuppressants, and biological agents, but there are problems of limited efficacy and large side effects. This example aims to evaluate the efficacy and safety of the HDAC6 and HSP90 dual-function inhibitors of the present invention in the treatment of UC.

[0060] 2. Treatment options

[0061] Thirty-six male C57BL / 6 mice were selected and randomly divided into six groups, with 6 mice in each group. The six groups were: blank control group (i.e., NC group, healthy mouse group), model group (i.e., DSS group, group with modeling but no drug treatment), 5-aminosalicylic acid group (i.e., 5-ASA group, 5-aminosalicylic acid was orally administered at a dose of 200 mg / kg, twice a day), low-dose administration group (compound 2 was orally administered at a dose of 5 mg / kg, twice a day), medium-dose administration group (compound 2 was orally administered at a dose of 10 mg / kg, twice a day), and high-dose administration group (compound 2 was orally administered at a dose of 20 mg / kg, twice a day). Each group was treated for 7 consecutive days.

[0062] 3. Treatment process

[0063] During the treatment period, the weight and condition of the mice were observed and recorded daily, and any adverse reactions were recorded.

[0064] 4. Experimental Results

[0065] After the treatment, the colons of mice in each group were removed for analysis. Compared with the DSS model group, the bloody stool symptoms of mice in the drug group were significantly improved, the number of diarrhea episodes was reduced by more than 80% compared with the model group, and the inflammatory area under colonoscopy was significantly reduced. Inflammatory indicators returned to normal ranges. In addition, the incidence of adverse reactions in the drug group was lower than that in the control group, no serious side effects were observed, and no mice died.

[0066] This example shows that the use of HDAC6 and HSP90 dual-function inhibitors in ulcerative colitis has significant therapeutic effects, reduces inflammatory responses, and has good safety. The dual-function inhibitors enhance the anti-inflammatory effect through synergistic effects, reduce the risk of drug resistance and side effects, and provide a new and effective means for the treatment of UC.

[0067] Example 2: Use of HDAC6 and HSP90 dual-function inhibitors in patients with Crohn's disease

[0068] 1. Experimental Background

[0069] Crohn's disease (CD) is a chronic inflammatory disease involving the entire digestive tract, often causing abdominal pain, weight loss and systemic symptoms. Although existing treatments can relieve symptoms, long-term use has drug resistance and side effects. This example aims to verify the effectiveness and safety of the HDAC6 and HSP90 dual-function inhibitor of the present invention in the treatment of CD.

[0070] 2. Treatment options

[0071] Twenty-four male C57BL / 6 mice were randomly divided into four groups, with 6 mice in each group. The six groups were: blank control group (i.e., NC group, healthy mouse group), model group, 5-aminosalicylic acid group (i.e., 5-ASA group, 5-aminosalicylic acid was orally administered at a dose of 200 mg / kg, twice a day), and compound administration group (compound 2 was orally administered at a dose of 20 mg / kg, twice a day). Each group was treated for 7 consecutive days.

[0072] 3. Treatment process

[0073] During the treatment period, the weight and condition of the mice were observed and recorded daily, and any adverse reactions were recorded.

[0074] 4. Experimental Results

[0075] After the treatment, the serum of the experimental mice was extracted to test the inflammatory indicators, and the results showed that the inflammatory indicators basically returned to normal. The incidence of adverse reactions in the experimental group was lower than that in the control group, and no mice died.

[0076] This example shows that the use of HDAC6 and HSP90 dual-function inhibitors in patients with Crohn's disease can significantly improve clinical efficacy, promote the relief of intestinal inflammation and ulcer healing, and have good safety. The dual-function inhibitor enhances anti-inflammatory and tissue repair capabilities through multi-target synergy, reduces drug resistance and adverse reactions, and provides an effective new treatment option for the treatment of Crohn's disease.

[0077] Through the above two specific examples, it can be seen that the application of the HDAC6 and HSP90 bifunctional inhibitor of the present invention in inflammatory bowel disease (including ulcerative colitis and Crohn's disease) shows significant therapeutic effect and good safety. The bifunctional inhibitor enhances the anti-inflammatory effect, promotes tissue repair, reduces drug resistance and side effects through the synergistic effect of dual targets, and has broad clinical application prospects and significant technical advantages.

[0078] The above description is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any modifications, equivalent substitutions and improvements made by any technician familiar with the technical field within the technical scope disclosed by the present invention and within the spirit and principle of the present invention should be covered by the protection scope of the present invention.

Claims

1. A dual-function inhibitor of HDAC6 and HSP90 for treating inflammatory bowel disease, characterized in that: The inhibitors include compounds having the following general formula: Wherein, R represents one of the following molecular formulas:

2. The HDAC6 and HSP90 dual-function inhibitor for treating inflammatory bowel disease according to claim 1, characterized in that: The inhibitor is a representative compound 1, whose structural formula is:

3. A method for using a dual-function inhibitor of HDAC6 and HSP90 for treating inflammatory bowel disease, characterized in that: The method comprises the following steps: Step S1, drug preparation: synthesizing the HDAC6 and HSP90 dual-function inhibitor according to claim 1 or 2; Step S2, selection of administration route: preparing the inhibitor into a pharmaceutical preparation suitable for oral administration, injection or other administration routes.

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