Use of a small molecule compound KA-C1 in the preparation of a medicament

Abstract

The application belongs to the field of medicine, and specifically discloses application of a small molecule compound KA-C1 in preparation of a medicine for preventing or treating cardiac hypertrophy, cardiac fibrosis and / or heart failure. The application finds that kynurenine-3-monooxygenase (KMO) is an effective medicine target for treating pathological cardiac hypertrophy and cardiac fibrosis through research. Through computer-aided virtual screening, the application finds a KMO-specific agonist KA-C1, and KA-C1, as a medicine active ingredient, can directly stimulate KMO protein activity, thereby effectively resisting pathological cardiac hypertrophy and cardiac fibrosis, and further preventing or treating heart failure. Therefore, the discovery of the small molecule compound KA-C1 provides a new medicine target strategy for treating pathological cardiac hypertrophy, cardiac fibrosis and heart failure, and has important and wide application prospects.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, specifically to the use of a small molecule compound KA-C1 in the preparation of medicaments for the prevention or treatment of myocardial hypertrophy and remodeling, and for inhibiting cardiac fibrosis and / or heart failure. This invention also relates to kynurenine-3-monooxygenase (KMO) agonists comprising this small molecule compound. Background Technology

[0002] Heart failure is a clinical syndrome whose incidence continues to rise with the increasing aging of the global population, making it a major public health problem threatening human health. While advancements in heart failure medications, cardiac device therapy, and treatment strategies have improved clinical symptoms and prognosis to some extent, readmission and mortality rates remain high. Therefore, further exploration of the pathological and molecular biological mechanisms of heart failure development and the identification of new intervention targets will contribute to the development of new drugs and improve the quality of life and prognosis of heart failure patients. Recent studies have revealed that the tryptophan metabolism pathway is a signaling molecule closely related to the regulation of energy expenditure and various metabolic diseases. Among these, the kynurenine (KYN) pathway, as a core pathway of tryptophan metabolism, plays a crucial role in the occurrence and progression of cardiovascular diseases. Recent metabolomics studies have found that kynurenine is associated with an increased risk of heart failure and even, to some extent, the risk of atrial fibrillation. Currently, there are no drugs in clinical practice that target kynurenine metabolism to treat heart failure. Summary of the Invention

[0003] Our team discovered that overexpression of KMO (a key metabolic enzyme of kynurenine) can effectively inhibit pathological myocardial hypertrophy, cardiac fibrosis, and heart failure. Based on this, developing small-molecule KMO agonists may serve as a strategy for the prevention and treatment of heart failure. KA-C1 is a KMO-specific small-molecule agonist that our team identified through high-throughput screening using virtual screening methods in computer-aided drug design. It exhibits excellent activity in inhibiting pathological myocardial hypertrophy to prevent or treat heart failure. It can directly bind to the KMO protein, promoting kynurenine metabolism, thereby regulating the AhR / BMP10 pathway to inhibit pathological myocardial hypertrophy / remodeling and achieve the goal of treating heart failure. Currently, there is a lack of drugs in clinical practice that target kynurenine metabolism to treat heart failure. This invention, on the one hand, reveals the target and mechanism of KA-C1 in inhibiting pathological myocardial hypertrophy and remodeling, and cardiac fibrosis; on the other hand, it provides new drugs and theoretical basis for the treatment of heart failure.

[0004] Based on this, the present invention aims to provide a new treatment strategy that reverses ventricular remodeling by targeting the KMO (key metabolic enzyme of kynurenine) pathway, thereby treating heart failure and improving the prognosis of heart failure patients.

[0005] To achieve the above objectives, the specific technical solution adopted by the present invention is as follows:

[0006] In a first aspect, the present invention provides the use of a small molecule compound KA-C1 in the preparation of a medicament for the prevention or treatment of myocardial hypertrophy, cardiac fibrosis, and / or heart failure, wherein the molecular structure of KA-C1 is as follows:

[0007]

[0008] Furthermore, the KA-C1 regulates kynurenine metabolism and stimulates KMO protein activity by specifically targeting kynurenine-3-monooxygenase (KMO), thereby effectively resisting pathological myocardial hypertrophy / remodeling and cardiac fibrosis, and thus preventing or treating heart failure.

[0009] Furthermore, the drug contains an effective amount of KA-C1 and pharmaceutically acceptable excipients.

[0010] Furthermore, the acceptable excipients include, but are not limited to, one or more of the following: diluents, excipients, fillers, binders, humectants, disintegrants, absorption promoters, surfactants, adsorbent carriers, lubricants, and 0.9% sodium chloride. Examples include: diluents and excipients such as water; fillers such as starch and sucrose; binders such as cellulose derivatives, alginate, gelatin, and polyvinylpyrrolidone; humectants such as glycerin; disintegrants such as agar, calcium carbonate, and sodium bicarbonate; absorption promoters such as quaternary ammonium compounds; surfactants such as hexadecyl alcohol; adsorbent carriers such as kaolin and soap clay; and lubricants such as talc, calcium / magnesium stearate, and polyethylene glycol.

[0011] Furthermore, the drug can be in dosage forms such as injections, capsules (including sustained-release or delayed-release forms), tablets, pills, suspensions, granules, tinctures, syrups, emulsions, and suspensions, as well as various sustained-release dosage forms, thus being suitable for various routes of administration, such as oral, parenteral, mucosal, intramuscular, intravenous, subcutaneous, intraocular, intradermal, or transdermal administration. Preferred are injectable injections or oral capsules; the KA-C1 injection dose (injection) is 10-20 mg / kg / day, and the KA-C1 oral dose (capsule) is 25-50 mg / kg / day.

[0012] Secondly, this invention provides the application of KMO as a target in the preparation of drugs for the prevention or treatment of myocardial hypertrophy and remodeling, and the inhibition of cardiac fibrosis and / or heart failure. Overexpression of the KMO gene can inhibit pathological myocardial hypertrophy and remodeling, improve cardiac fibrosis, and prevent or treat heart failure.

[0013] Thirdly, this invention provides the application of the aforementioned KA-C1 in the preparation of KMO small molecule agonists. The agonist contains KA-C1, which can directly bind to KMO proteins; by targeting KMO, it can effectively inhibit myocardial hypertrophy / remodeling and cardiac fibrosis, promote kynurenine metabolism, and prevent or treat heart failure.

[0014] Furthermore, the KMO small molecule agonist comprises KA-C1 and the aforementioned pharmaceutically acceptable excipients.

[0015] Fourthly, the present invention provides a pharmaceutical composition comprising an effective amount of KA-C1 and the aforementioned pharmaceutically acceptable excipients.

[0016] The present invention has the following beneficial effects:

[0017] For the treatment of heart failure, this invention has discovered that kynurenine-3-monooxygenase (KMO) can serve as an effective drug target for treating pathological myocardial hypertrophy and myocardial fibrosis. Furthermore, KA-C1, a KMO-specific small molecule agonist, has been developed for the treatment of heart failure. KA-C1 can directly bind to and activate the KMO protein, thereby inhibiting pathological myocardial hypertrophy and cardiac fibrosis, achieving the effect of preventing or treating heart failure. It also shows significant efficacy as a drug for reversing ventricular remodeling and treating heart failure. Therefore, this invention provides a novel drug target strategy for the treatment of pathological myocardial hypertrophy, cardiac fibrosis, and heart failure, with significant and broad application prospects. Attached Figure Description

[0018] Figure 1 This study investigates the effects of systemic KMO knockout on Ang II-induced pathological myocardial hypertrophy and cardiac function in mice. In the figures: A shows echocardiographic images of mice; B shows the statistical results of ejection fraction (EF%) detected by echocardiography; C shows the statistical results of fractional shortening (FS%) detected by echocardiography; D shows a representative gross image of the heart; E shows the statistical results of cardiac cross-sectional area; F shows H&E staining images; G shows WGA staining images; H shows the statistical results of cardiomyocyte cross-sectional area; I shows Western blot images of the myocardial hypertrophy markers ANP and β-MyhC; J shows the quantitative statistical results of protein expression levels detected by Western blot.

[0019] Figure 2The effect of systemic KMO knockout on Ang II-induced myocardial fibrosis in mice. In the figure: A is a Masson staining image; B is a representative Sirius Red staining image; C shows the statistical results of the percentage of cardiac collagen content stained by Masson and Sirius Red staining; D shows Western Blot images of fibrosis markers Col-I and TGF-β; E shows the quantitative statistical results of protein expression levels detected by Western Blot. n = 6, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

[0020] Figure 3 This is the molecular structural formula of the small molecule compound KA-C1 described in this invention.

[0021] Figure 4 This is a graph showing the analytical results of the binding of the small molecule compound KA-C1 described in this invention to the KMO protein.

[0022] Figure 5 This invention illustrates the effects of the small molecule compound KA-C1 on Ang II-induced pathological myocardial hypertrophy and cardiac function in mice. In the figure: A is an echocardiogram image; B is the statistical result of EF% from echocardiography; C is the statistical result of FS% from echocardiography; D is a representative gross image of the heart; E is the statistical result of cardiac cross-sectional area; F is an H&E staining image; G is a WGA staining image; H is the statistical result of cardiomyocyte cross-sectional area; I is a Western blot image of ANP and β-MyhC; J is the quantitative statistical result of protein expression levels detected by Western blot.

[0023] Figure 6 This invention illustrates the effect of the small molecule compound KA-C1 on Ang II-induced myocardial fibrosis in mice. In the figure: A is a Masson staining image; B is a representative Sirius Red staining image; C shows the statistical results of the percentage of cardiac collagen content stained by Masson and Sirius Red staining; D shows the Western Blot images of fibrosis markers Col-I and TGF-β; E shows the quantitative statistical results of protein expression levels detected by Western Blot. n = 6, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

[0024] Figure 7This invention relates to the effects of the small molecule compound KA-C1 on cardiac function and ventricular remodeling in mice with myocardial infarction-induced myocardial hypertrophy. In the figure: A is an echocardiogram image; B is the statistical result of EF% from echocardiogram; C is the statistical result of FS% from echocardiogram; D is the result of ELISA detection of serum ANP levels; E is a representative gross image of the heart; F is an H&E staining image; G is a Masson staining image; H is the statistical result of the percentage of collagen content in the heart stained by Masson staining; I is a SiriusRed staining image; J is the statistical result of the percentage of collagen content in the heart stained by SiriusRed staining. n = 6, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Detailed Implementation

[0025] The specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings and examples.

[0026] Experimental animals: Wild-type C57BL / 6 mice (WT) and KMO gene knockout C57BL / 6 mice (KMO) were used in this invention. - / - All of these were from Nanjing Jicui Yaokang Biotechnology Co., Ltd., and all animal care and experimental procedures were approved by the ethics committees of Fudan University Affiliated Zhongshan Hospital and Wenzhou Medical University.

[0027] In Examples 1-3, an Ang II-induced myocardial hypertrophy model was established by implanting an Ang II sustained-release pump (Ang II final concentration of 1 μg / kg / min, pump release rate of 0.11 μL / h) in the back of mice for a total of 28 days. In Example 4, a mouse model of myocardial infarction (MI)-induced ventricular remodeling was established by permanently ligating the origin of the left anterior descending artery (LAD) 2 mm with 7-0 surgical sutures under isoflurane-induced anesthesia to simulate clinical myocardial infarction.

[0028] Example 1: Investigation on the effects of KMO on cardiac function and ventricular remodeling in Ang II-induced myocardial hypertrophy mice

[0029] KMOs of similar weight and age (8 weeks) were selected. - / - Mice and their corresponding wild-type mice were randomly divided into 4 groups of 6 mice each, with 1 KMO mouse selected from each group. - / - An Ang II-induced myocardial hypertrophy model was established using mice and corresponding wild-type mice (denoted as KMO). - / - +Ang II, WT+Ang II group), synchronously set KMO - / - Mice and a sham-operated group of corresponding wild-type mice (mice with a slow-release saline pump implanted in their backs) served as controls (denoted as KMOs). - / -+Sham, WT+Sham group).

[0030] Results of cardiac function detection in mice using ultra-high resolution small animal ultrasound, such as Figure 1 As shown in AC, knocking out KMO - / - KMO (Kardiomyopathy) model mice - / - The ejection fraction (EF%) and fractional shortening (FS%) of Ang II-induced mice were significantly decreased, indicating that KMO knockout significantly aggravated Ang II-induced cardiac dysfunction in mice. H&E staining and WGA staining were used to observe myocardial morphology and cardiomyocyte hypertrophy in mice, and the results were as follows: Figure 1 As shown in the DH diagram, KMO knockout significantly aggravated Ang II-induced myocardial morphological disturbances and cardiomyocyte hypertrophy in mice. Furthermore, the protein levels of myocardial hypertrophy markers (β-MyhC, ANP) were detected by Western blot, and the results are as follows: Figure 1 As shown in IJ, KMO - / - The levels of myocardial hypertrophy marker proteins were significantly increased in the +Ang II group, and KMO knockout significantly aggravated Ang II-induced myocardial hypertrophy in mice.

[0031] Meanwhile, the fibrosis of mouse myocardial tissue was observed using Masson and Sirius staining, and the results were as follows: Figure 2 As shown in AC, KMO knockout significantly increased Ang II-induced myocardial collagen fiber deposition; protein levels of fibrosis markers (Col-I, TGF-β) were detected by Western blot, and the results are as follows. Figure 2 As shown in DE, KMO - / - The levels of fibrosis marker proteins were significantly increased in the Ang II group. Combined with these results, we found that KMO knockout exacerbated Ang II-induced myocardial fibrosis in mice.

[0032] Based on the above results, we found that systemic KMO knockout significantly aggravated Ang II-induced cardiac dysfunction, myocardial hypertrophy, and cardiac fibrosis.

[0033] Example 2: Evaluation of the binding ability of KA-C1 to KMO protein

[0034] This embodiment utilizes SPR (Surface Plasmon Resonance) for biomolecular interaction analysis, employing Biacore. TM The molecular interaction analysis system, Cytiva's fully automated testing platform, is used to assess drug-protein binding relationships. Specifically, it uses SPR analysis of KA-C1 (structural formula as shown in the image). Figure 3(As shown) the binding interaction with KMO protein. We diluted the purified KMO protein to 100 μg / mL with PBS at pH 5.5 and coupled the KMO protein to the blank CM5 chip in 10 mM sodium acetate buffer; we serially diluted KA-C1 with 1‰ DMSO+PBS to 5 concentrations, with the highest concentration being 50 μM and the lowest concentration being 3.125 μM, for subsequent detection; after adding the above samples to the sample plate in sequence, we ran the program using Biacore. TM The software performs data fitting for dynamic and steady-state analysis, and finally we perform fitting analysis on the obtained data using a 1:1 fitting model.

[0035] The results are as follows Figure 4 As shown, there is a strong interaction between KA-C1 and KMO protein (dissociation constant KD = 7.27 μM). This result indicates that KA-C1 can directly bind to KMO protein and is kinetically stable.

[0036] Example 3: Investigation on the effects of KA-C1 on cardiac function and ventricular remodeling in Ang II-induced myocardial hypertrophy mice

[0037] Healthy, age-matched (8 weeks old) C57BL / 6 mice with similar weights were randomly divided into two groups of 6 mice each by blind selection. While establishing the Ang II-induced myocardial hypertrophy model, the mice were given either KA-C1 (KA-C1 from Enamine Ltd, with an effective concentration of 10 mg / kg / d, dissolved in 25 mg / ml stock solution using DMSO and then evenly distributed in sesame oil, and the DMSO solution containing KA-C1 was safely delivered into the mice to promote KA-C1 absorption, administered by gavage every other day) or an equal amount of sesame oil, respectively. These groups were designated as KMO activator-1+Ang II (or KA-C1+Ang II) and WT+Ang II groups.

[0038] The cardiac function of mice was detected using ultra-high resolution small animal ultrasound, and the results are as follows: Figure 5 As shown in AC, KA-C1 increased the ejection fraction (EF%) and fractional shortening (FS%) in Ang II-induced myocardial hypertrophy mice, significantly improving Ang II-induced cardiac dysfunction; and the morphology of mouse myocardium and cardiomyocyte hypertrophy were observed by H&E staining and WGA staining, with results as follows. Figure 5 As shown in the DH, KA-C1 significantly improved Ang II-induced myocardial morphological disturbances and cardiomyocyte hypertrophy in mice. Furthermore, the protein levels of myocardial hypertrophy markers (β-MyhC, ANP) were detected by Western blot, and the results are as follows: Figure 5As shown in IJ, the levels of myocardial hypertrophy marker proteins were significantly decreased in the KMOactivator-1+Ang II group, and KA-C1 significantly improved Ang II-induced myocardial hypertrophy in mice. Combining these results, we found that KA-C1 can significantly improve Ang II-induced cardiac dysfunction and myocardial hypertrophy.

[0039] Meanwhile, the fibrosis of mouse myocardial tissue was observed using Masson and Sirius staining, and the results were as follows: Figure 6 As shown in AC, KA-C1 significantly reduced Ang II-induced myocardial collagen fiber deposition; the protein levels of fibrosis markers (Col-I, TGF-β) were detected by Western blot, and the results are as follows. Figure 6 As shown in the DE, the levels of fibrosis marker proteins in the KA-C1+Ang II group were significantly decreased, and KA-C1 significantly reduced the level of myocardial fibrin in Ang II mice. Combining these results, we found that KA-C1 can significantly improve the level of myocardial fibrosis induced by Ang II in mice.

[0040] Example 4: Investigation on the effects of KA-C1 on cardiac function and ventricular remodeling in mice induced by myocardial infarction

[0041] Healthy, age-matched (8 weeks old) C57BL / 6 mice with similar weights were randomly divided into two groups of 6 mice each by blind selection. After establishing a mouse model of myocardial infarction (MI)-induced ventricular remodeling, the mice were given KA-C1 (10 mg / kg / d, dissolved in DMSO to a stock solution of 25 mg / ml and evenly distributed in sesame oil, administered by gavage every other day for 4 weeks) or an equal amount of sesame oil, which were designated as KMOactivator-1+MI group and WT+MI group, respectively.

[0042] The cardiac function of mice was detected using ultra-high resolution small animal ultrasound, and the results are as follows: Figure 7 As shown in AC, KA-C1 increased ejection fraction (EF%) and fractional shortening (FS%) in mice with MI-induced ventricular remodeling, significantly improving MI-induced cardiac dysfunction; serum atrial natriuretic peptide (ANP) levels in mice were detected using ELISA, and the results are as follows. Figure 7 As shown in Figure D, KA-C1 significantly reduced serum ANP levels in MI mice, suggesting improved cardiac function. Simultaneously, H&E staining was used to observe myocardial morphology and cardiomyocyte remodeling in mice, with results as follows: Figure 7 As shown in EF, KA-C1 significantly improved MI-induced myocardial morphological disturbances and ventricular remodeling in mice. These results indicate that KA-C1 can significantly improve MI-induced cardiac dysfunction and ventricular remodeling. Simultaneously, Masson and Sirius staining were used to observe fibrosis in mouse myocardial tissue, and the results are as follows... Figure 7As shown in GJ, KA-C1 significantly reduced MI-induced myocardial collagen fiber deposition. We found that KA-C1 can significantly improve the level of MI-induced myocardial fibrosis in mice.

[0043] In summary, this invention obtained a potential KMO agonist, KA-C1, through virtual screening of existing commercial compound libraries. We confirmed through biomolecular interaction analysis using the surface plasmon resonance (SPR) principle that KA-C1 can directly bind to KMO proteins. In mouse models of pathological myocardial hypertrophy and myocardial infarction, KA-C1 effectively alleviated myocardial hypertrophy and fibrosis, reversed ventricular remodeling, and improved cardiac function. These results indicate that KA-C1 specifically targets KMOs to improve pathological myocardial hypertrophy and remodeling, inhibit myocardial fibrosis, and thus alleviate heart failure.

[0044] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the design concept of the present invention should be included within the protection scope of the present invention.

Claims

1. The use of a small molecule compound KA-C1 in the preparation of a medicament for the prevention or treatment of myocardial hypertrophy, cardiac fibrosis, and / or heart failure, wherein the molecular structure of KA-C1 is as follows: 。 2. The application according to claim 1, characterized in that, The KA-C1 specifically targets kynurenine-3-monooxygenase (KMO), regulates kynurenine metabolism, and stimulates KMO protein activity, thereby effectively resisting pathological myocardial hypertrophy and remodeling, inhibiting cardiac fibrosis, and thus preventing or treating heart failure.

3. The application according to claim 1, characterized in that, The drug contains an effective amount of KA-C1 and pharmaceutically acceptable excipients.

4. In the application according to claim 3, the pharmaceutically acceptable excipient is selected from one or more of diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorbents, lubricants, and 0.9% sodium chloride.

5. The application according to claim 3, characterized in that, The dosage form of the drug is injection, capsule, tablet, pill, suspension, granule, tincture, syrup, emulsion or suspension.

6. The application according to claim 5, characterized in that, The drug is available in the form of an injection or an oral capsule; the injection dose of KA-C1 is 10-20 mg / kg / day, and the oral dose is 25-50 mg / kg / day.

7. The use of KMO as a target in the preparation of drugs for the prevention or treatment of myocardial hypertrophy, cardiac fibrosis and / or heart failure.

8. Application of KA-C1 in the preparation of KMO small molecule agonists, wherein the molecular structure of KA-C1 is as follows: 。 9. The application according to claim 8, characterized in that, The KMO small molecule agonist comprises KA-C1 and pharmaceutically acceptable excipients.

10. A pharmaceutical composition, characterized in that, It contains an effective amount of KA-C1 and pharmaceutically acceptable excipients.

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