WO2022116583A1 - Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用 - Google Patents

Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用 Download PDF

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WO2022116583A1
WO2022116583A1 PCT/CN2021/111105 CN2021111105W WO2022116583A1 WO 2022116583 A1 WO2022116583 A1 WO 2022116583A1 CN 2021111105 W CN2021111105 W CN 2021111105W WO 2022116583 A1 WO2022116583 A1 WO 2022116583A1
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wnt
dilated cardiomyopathy
scn5a
preparation
mutation
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PCT/CN2021/111105
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French (fr)
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葛均波
孙爱军
胡静静
杨坤
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复旦大学附属中山医院
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Priority to EP21844611.0A priority Critical patent/EP4101451A4/en
Priority to US17/637,455 priority patent/US11779575B2/en
Priority to JP2022508833A priority patent/JP7297146B2/ja
Publication of WO2022116583A1 publication Critical patent/WO2022116583A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • the invention relates to the technical field of biomedicine, in particular to the application of Wnt inhibitor Wnt-C59 in preparing a medicine for treating dilated cardiomyopathy caused by SCN5A mutation.
  • dilated cardiomyopathy refers to irreversible cardiac function decline caused by cardiac enlargement.
  • the population prevalence is about 40/100,000 people, and the 5-year mortality rate is 50%. There is no specific treatment plan.
  • About 20-30% of cardiomyopathy is caused by genetic factors. Initially, it was found that its pathogenic genes encode multiple cellular structural proteins, such as cardiac actin (actin) gene, nuclear lamin (laminA/C) gene, etc. Cellular stress transmission leads to myocardial damage and cardiac enlargement.
  • actin actin
  • laminA/C nuclear lamin
  • the functional unit of the cardiac sodium channel encoded by the SCN5A gene is composed of 4 homologous regions (DI-DIV), each of which is composed of 6 transmembrane segments S1-S6.
  • the S4 segment is rich in positively charged residues and is a channel protein voltage receptors.
  • both R225Q and A226V are located in the S4 segment of the DI region, that is, 80% (4/5) of the SCN5A mutations found are located in the voltage receptor site of the channel, except for 2 synonymous mutations.
  • Previous studies have reported that dilated cardiomyopathy-related SCN5A mutations R814W, T220I, etc. are also located in this site.
  • Struyk has reported that mutations in the voltage receptor site of skeletal muscle sodium channels can produce a gating pore effect, resulting in abnormal hydrogen ion leakage, and persistent leakage can cause abnormal skeletal muscle function by interfering with intracellular pH.
  • SCN5A mutations A1180V, R225Q, A226V, I1448N, I94 and Y1434 through candidate gene sequencing analysis of the family with cardiomyopathy, and confirmed that a series of SCN5A mutants are related to cardiomyopathy.
  • the pathogenesis and clinical application are still poorly understood. Based on this, we put forward the hypothesis that mutations in the voltage receptor site of the cardiac sodium channel gene lead to unique abnormal currents, causing dysregulation of the intracellular environment (such as changes in pH and Ca2+ balance), resulting in myocardial structural damage and dysfunction.
  • Wnt signaling pathway plays a key role in the regulation of cardiovascular development and cardiac hypertrophy.
  • the Wnt signaling pathway is particularly active during cardiac development, whereas in the normal adult mammalian heart, Wnt signaling is quiescent, but it is activated during cardiac pathology.
  • Wnt signaling is involved in the process of cardiac ventricular remodeling, cardiac hypertrophy, and heart failure.
  • differentially expressed mRNAs were significantly enriched in the Wnt signaling pathway. It can be inferred that the occurrence and development of DCM may be related to the activation of the Wnt signaling pathway. Intervention of this pathway may provide new targets for the treatment of DCM.
  • the present invention proposes for the first time that Wnt inhibitor Wnt-C59 can be used to treat dilated cardiomyopathy caused by SCN5A mutation. There is no report about the application of the Wnt inhibitor Wnt-C59 of the present invention in preparing a medicine for treating dilated cardiomyopathy caused by SCN5A mutation.
  • the purpose of the present invention is to provide a new drug for treating dilated cardiomyopathy caused by SCN5A mutation in view of the deficiencies of the prior art.
  • the present invention provides the application of a Wnt inhibitor in the preparation of a medicament for treating dilated cardiomyopathy caused by SCN5A mutation.
  • the present invention provides the application of the Wnt inhibitor Wnt-C59 in the preparation of a medicine for treating dilated cardiomyopathy caused by SCN5A mutation.
  • the present invention provides the application of the Wnt inhibitor Wnt-C59 in the preparation of a medicine for treating dilated cardiomyopathy caused by the SCN5A-R225Q mutant.
  • the present invention provides the application of the Wnt inhibitor Wnt-C59 in the preparation of a medicament for improving the cardiac function and prognosis of patients with dilated cardiomyopathy caused by the SCN5A-R225Q mutant.
  • the present invention provides the application of Wnt inhibitor Wnt-C59 in the preparation of a medicine for improving ventricular cavity enlargement, myocardial contractile function decline and prognosis in patients with dilated cardiomyopathy caused by SCN5A-R225Q mutant.
  • the above-mentioned pharmaceutical dosage forms are selected from tablets, powders, suspensions, capsules, pills or syrups.
  • dilated cardiomyopathy caused by different gene mutations will have obvious individual differences.
  • the genes causing dilated cardiomyopathy also include MYH7, KCNQ1, ABCC9, CLIC2, etc.
  • Symptoms such as chest tightness, shortness of breath, lower extremity edema, decreased exercise tolerance, dyspnea, etc., can also occur atrioventricular block, sinus bradycardia, sinus arrest, ventricular premature beats, ventricular tachycardia, complete bundle branch
  • Electrocardiographic changes such as conduction block, symptoms such as dizziness, fatigue, syncope, palpitations, etc., and their clinical treatment plans are also different.
  • SCN5A is a gene encoding Na+ channel, so the typical features of dilated heart failure are arrhythmia, and common symptoms are palpitations, dizziness, fatigue, syncope, etc.
  • MYH7 is a gene encoding myosin, so the typical manifestation of dilated cardiomyopathy is myocardial Mainly decreased contractility, clinical manifestations of chest tightness, shortness of breath, dyspnea, lower extremity edema, decreased activity tolerance and so on.
  • the clinical symptoms, pathological features, and treatment methods of cardiomyopathy caused by different site mutations are also significantly different, as follows:
  • Mutations at different sites can cause different types of arrhythmias, such as atrioventricular block, sinus bradycardia, sinus arrest, premature ventricular contractions, ventricular tachycardia, complete bundle branch block, etc.
  • Symptoms include dizziness, fatigue, syncope, and palpitations, and their clinical treatment options are also different.
  • Pathological features will also show different changes due to different mutation sites, such as the pathological manifestations of dilated cardiomyopathy dominated by cardiac interstitial fibrosis, the pathological manifestations of cardiomyocyte hypertrophy, and inflammatory cell infiltration, etc.;
  • CRT cardiac resynchronization therapy
  • bradyarrhythmias such as atrioventricular block, sinus bradycardia, sinus arrest, etc.
  • a temporary pacemaker or permanent dual-phase pacemaker on the basis of standard treatment and according to the severity of the patient's symptoms.
  • Chamber/single-chamber pacemaker implantation if the patient has tachyarrhythmias, such as ventricular premature beats, ventricular tachycardia, etc., radiofrequency ablation or implantation of an ICD (implantable cardiac defibrillator) is required Wait.
  • ICD implantable cardiac defibrillator
  • Wnt-C59 is an inhibitor of Wnt protein activation and can block the Wnt signaling pathway from the beginning. In tumor treatment experiments, Wnt-C59 can significantly inhibit the proliferation of 46 tumor cells. In this study, we used aging combined with doxorubicin-induced cardiomyopathy model, and detected the therapeutic effect of Wnt-C59 on cardiomyopathy through changes in cardiac function and activation of related signaling molecules. Type cardiomyopathy provides a theoretical basis.
  • the present invention discovers for the first time that the dilated cardiomyopathy caused by the SCN5A-R225Q mutant is related to the Wnt signaling pathway. Inhibit the abnormal activation of Wnt/ ⁇ -Catenin pathway caused by SCN5A gene mutation, so as to improve the prognosis of cardiac function in patients with dilated cardiomyopathy with SCN5A gene mutation, and provide a theoretical basis for the application of Wnt-C59 in clinical treatment of dilated cardiomyopathy. It provides a new target for the treatment of dilated cardiomyopathy and a new treatment method for this type of patients, which can be well applied in clinic and has a good application prospect.
  • Figure 1 shows the results of LVEF index of mice in each group. The results show that the cardiac function of mice in the KI+Wnt-C59 group was significantly improved after gavage with Wnt-C59 (*: P ⁇ 0.05; ***: P ⁇ 0.001).
  • Figure 2 shows the results of M-mode images of mice in each group. The results show that the cardiac function of mice in the KI+Wnt-C59 group was significantly improved after intragastric administration of Wnt-C59 (*: P ⁇ 0.05; ***: P ⁇ 0.001).
  • Figure 3 shows the results of Western blot detection of mice in each group. The results showed that the Wnt pathway was significantly activated in the KI group compared with the WT group.
  • Figure 4 shows the size of the hearts of the mice in each group. The results showed that the hearts of the mice in the KI+Wnt-C59 group were significantly smaller than those in the KI group.
  • Doxorubicin model was established while the KI+Wnt-C59 group was intragastrically administered with Wnt-C59 (1 mg/kg.d, MCE HY-15659) every day, and the WT and KI groups were intragastrically administered with an equal volume of sterile normal saline as a control.
  • Echocardiography was detected 2 weeks after administration with a probe frequency of 30MHz. After specifically isoflurane gassing the animals, M-mode images were recorded while the mouse's heart rate was maintained at 450-500 beats/min. B-Mode images of the parasternal long-axis view and the apical four-chamber view were collected. The parasternal short-axis left ventricle was taken, 2D ultrasonography showed the short-axis view of the left ventricle, and M-mode ultrasonography was used to record the motion of the left ventricle at the level of the papillary muscle. Functional indicators include: left ventricular ejection fraction (LVEF). The morphological and functional changes of the hearts of the mice in each group were compared. All measurements were the average of 5 consecutive cardiac cycles and were performed by 3 experienced technicians.
  • LVEF left ventricular ejection fraction
  • the cardiomyocyte proteins of KI group and WT group were extracted and detected by Western blot.
  • mice were sacrificed after 2 weeks of drug intervention, and the hearts were taken to compare the heart size among the three groups.
  • Wnt signaling is involved in cardiac ventricular remodeling, cardiac hypertrophy, and heart failure. Activation of Wnt signaling pathway is related to the occurrence and development of DCM. Based on our findings, Wnt signaling pathway inhibitor, Wnt-C59, can significantly improve the cardiac function in mice with cardiomyopathy induced by aging combined with doxorubicin, providing a theoretical basis for the clinical application of Wnt-C59 in the treatment of dilated cardiomyopathy, and also for the treatment of dilated cardiomyopathy. The treatment of dilated cardiomyopathy provides new targets.
  • Wnt-C59 can not only significantly inhibit the proliferation of 46 tumor cells, but also improve the cardiac function of mice with galactose senescence combined with doxorubicin-induced cardiomyopathy. It provides a theoretical basis for clinical treatment of dilated cardiomyopathy. Wnt-C59 is an important supplement to the previous traditional cardiotonic, diuretic, and vasodilator treatments for myocardial injury in cardiomyopathy. It can delay the progression of myocardial injury and improve the quality of life of patients.

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Abstract

本发明公开了Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用。本发明首次提出以SCN5A基因型检测为切入点，利用Wnt通路特异性抑制剂Wnt-C59抑制SCN5A基因突变所致的Wnt/β-Catenin通路异常活化，从而实现改善SCN5A基因突变的扩张型心肌病患者心脏功能预后。实验通过构建衰老联合阿霉素致扩心病模型，对心功能改变、相关信号分子的激活等指标检测Wnt-C59对扩心病的治疗作用，为Wnt-C59应用于临床治疗扩张型心肌病提供理论依据。本发明为SCN5A突变致扩张型心肌病提供了新的治疗方法。

Description

Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用 技术领域
本发明涉及生物医药技术领域，具体地说，是Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用。
背景技术
原发性扩张型心肌病(扩心病)指心脏扩大导致的不可逆心功能衰退，人群患病率约40/10万人，5年病死率达50％，尚无特异性治疗方案。由遗传因素导致的扩心病约占20-30％，最初发现其致病基因多编码细胞结构蛋白，如心肌肌动蛋白(actin)基因、核纤层蛋白(laminA/C)基因等，由于损害了细胞的应力传递而导致心肌损伤和心脏扩大。2004年McNair首次提出心脏钠通道基因(SCN5A)突变与扩心病有关。
SCN5A基因编码的心脏钠通道功能单位由4个同源区(DI～DIV)组成，每个同源区又由6个跨膜片段S1～S6构成，S4片段富含正电荷残基，是通道蛋白的电压感受器。有趣的是，R225Q和A226V均位于DI区的S4片段，也就是说，除外2个同义突变，所发现的SCN5A突变中80％(4/5)位于通道的电压感受器部位。既往有研究报道扩心病相关SCN5A突变R814W、T220I等也位于该部位。Struyk曾报道骨骼肌钠通道电压感受器部位的突变可产生门孔(gating pore)效应，导致异常氢离子泄漏，而持续泄露通过干扰细胞内pH值引起骨骼肌功能异常。我们前期研究通过对扩心病家系进行候选基因测序分析，发现了一系列新的SCN5A突变A1180V、R225Q、A226V、I1448N、I94和Y1434，并证实SCN5A一系列突变体与扩心病有关，然而其中的致病机制和临床应用仍不甚了解。基于此，我们提出假说：心脏钠通道基因电压感受器部位的突变导致独特的异常电流，引起细胞内环境的失调(如pH值、Ca2+平衡的变化)，从而导致心肌结构损伤和功能异常。
既往研究发现，Wnt信号通路在调节心脏血管发育及心肌肥大过程中起着关 键作用。Wnt信号通路在心脏发育期间特别活跃，而在正常成年哺乳动物心脏中，Wnt信号是静止的，但在心脏病理过程中Wnt信号通路被激活。研究发现Wnt信号参与心脏心室重构、心肌肥大、心力衰竭过程。相关研究进一步发现，差异表达的mRNAs在Wnt信号通路中显着富集，可以推断DCM的发生发展可能与Wnt信号通路的激活有关，干预这条途径可能为DCM的治疗提供新的靶点。在此基础上，我们证实，SCN5A-R225Q突变体致扩张型心肌病过程中，Wnt信号通路较野生型激活更加显著，提示Wnt信号通路在SCN5A-R225Q突变体致扩张型心肌病过程中扮演重要角色。本发明首次提出了Wnt抑制剂Wnt-C59可用于治疗SCN5A突变致扩张型心肌病。关于本发明Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用目前还未见报道。
发明内容
本发明的目的是针对现有技术的不足，提供了一种治疗SCN5A突变所致扩张型心肌病的新药物。
为实现上述目的，本发明采取的技术方案是：
第一方面，本发明提供了Wnt抑制剂在制备治疗SCN5A突变致扩张型心肌病的药物中的应用。
进一步地，本发明提供了Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用。
第二方面，本发明提供了Wnt抑制剂Wnt-C59在制备治疗SCN5A-R225Q突变体致扩张型心肌病的药物中的应用。
进一步地，本发明提供了Wnt抑制剂Wnt-C59在制备改善SCN5A-R225Q突变体致扩张型心肌病患者心脏功能及预后的药物中的应用。
更进一步地，本发明提供了Wnt抑制剂Wnt-C59在制备改善SCN5A-R225Q突变体致扩张型心肌病患者心室腔扩大、心肌收缩功能下降及预后的药物中的应用。
优选地，以上所述药物剂型均选自片剂、粉剂、混悬剂、胶囊剂、丸剂或糖浆。
本领域技术人员熟知，不同的基因突变引起扩心病的临床表现和心电图检查 会有明显个体差异，引起扩心病的基因除SCN5A，还有MYH7、KCNQ1、ABCC9、CLIC2等，患者除了心衰常见的症状如胸闷气急，下肢水肿，运动耐量下降，呼吸困难等，还可以出现房室传导阻滞、窦性心动过缓、窦性停搏、室性早搏、室性心动过速、完全性束支传导阻滞等心电图改变，症状上会有头晕乏力、晕厥、心悸等表现，其临床治疗方案也有不同。如：SCN5A是编码Na+通道的基因，因此扩心心衰典型特征以心律失常为主，常见症状有心悸、头晕乏力、晕厥等；MYH7是编码肌球蛋白的基因，因此扩心病典型表现以心肌收缩力下降为主，临床表现为胸闷气急、呼吸困难、下肢水肿、活动耐力下降等。
此外，不同的位点突变所致的扩心病在临床症状、病理特征、治疗方法上也有显著区别，具体如下：除了心衰常见的症状如胸闷气急，下肢水肿，运动耐量下降，呼吸困难等，不同位点的突变可以出现不同类型的心律失常，如房室传导阻滞、窦性心动过缓、窦性停搏、室性早搏、室性心动过速、完全性束支传导阻滞等，症状上会有头晕乏力、晕厥、心悸等表现，其临床治疗方案也有不同。病理特征也会因突变位点的不同呈现不同的改变，如心脏间质纤维化为主的扩心病病理表现、心肌细胞肥大、炎症细胞浸润为主的病理表现等等；
除常规强心、利尿、扩血管、改善心室重构等药物治疗，CRT(心脏再同步化治疗)目前也广泛应用于临床。如果患者出现缓慢性心律失常，如房室传导阻滞、窦性心动过缓、窦性停搏等，则需要在标准治疗的基础上，根据患者症状的轻重，选择临时起搏器或永久双腔/单腔起搏器植入；如果患者出现快速性心律失常，如室性早搏、室性心动过速等，则需要行射频消融术或植入ICD(可植入式心脏除颤仪)等。
Wnt-C59是Wnt蛋白活化的抑制剂，可从起始阻断Wnt信号通路。在肿瘤治疗实验中，Wnt-C59可以对46种肿瘤细胞的增殖产生显著的抑制作用。在本研究中，我们应用衰老联合阿霉素致扩心病模型，通过对心功能改变、相关信号分子的激活等指标检测Wnt-C59对扩心病的治疗作用，为Wnt-C59应用于临床治疗扩张型心肌病提供理论依据。
本发明优点在于：
本发明在传统标准化治疗的基础上，首次发现SCN5A-R225Q突变体所致的扩张型心肌病与Wnt信号通路有关，通过以SCN5A基因型检测为切入点，利用 Wnt通路特异性抑制剂Wnt-C59抑制SCN5A基因突变所致的Wnt/β-Catenin通路异常活化，从而实现改善SCN5A基因突变的扩张型心肌病患者心脏功能预后，为Wnt-C59应用于临床治疗扩张型心肌病提供理论依据，同时也为扩心病的治疗提供新的靶点，为该类患者提供了新的治疗方法，可很好的应用于临床上，具有很好的应用前景。
附图说明
附图1是各组小鼠LVEF指标结果，结果表明结果表明KI+Wnt-C59组灌胃Wnt-C59后小鼠心功能明显改善(*：P＜0.05；***：P＜0.001)。
附图2是各组小鼠M-mode图像结果，结果表明KI+Wnt-C59组灌胃Wnt-C59后小鼠心功能明显改善(*：P＜0.05；***：P＜0.001)。
附图3是各组小鼠Western blot检测结果，结果表明，KI组较WT组，Wnt通路明显激活。
附图4是各组小鼠心脏大小，结果表明：KI+Wnt-C59组小鼠心脏明显小于KI组。
具体实施方式
下面结合具体实施方式，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。此外应理解，在阅读了本发明记载的内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
本研究基于前期临床扩心病患者资料，即对90例散发扩心病进行SCN5A突变筛查，结果发现R225Q突变(c.674G＞A)率最高(共计3例)，且扩心病发病年龄偏大(分别为77，55，80岁)。
实施例1 动物实验
1方法
1.1实验动物及动物模型制备
选用SPF级雄性C57BL/6小鼠(购自上海杰思捷实验动物有限公司)和SCN5A杂合突变小鼠，周龄为8-10周，体重20-25g，用d-半乳糖(500mg/kg.d， sigma G5388)持续腹腔注射8周，构建衰老模型。然后用阿霉素(总量10mg/kg，分4次腹腔注射，生工A603456)，构建扩心模型。模型建立后分离KI组和WT组小鼠心肌细胞。实验方案得到复旦大学动物管理与伦理委员会批准。
动物实验证明，自然衰老模型、单纯半乳糖衰老模型和单纯阿霉素扩心模型，SCN5A杂合突变小鼠和野生型小鼠的心功能均无统计学差异。
前期动物研究发现，半乳糖+阿霉素处理后，SCN5A杂合突变小鼠出现扩心的表型，即心腔扩大，左室变薄，射血分数下降，而野生型小鼠心功能无变化。进一步分离KI组和WT组心肌细胞，两组心肌细胞Western blot显示，KI组Wnt通路明显激活。
1.2分组
小鼠测过超声后，研究共3组：WT组；KI组；KI+Wnt-C59组。阿霉素造模同时KI+Wnt-C59组每天灌胃Wnt-C59(1mg/kg.d，MCE HY-15659)，WT和KI组用等体积无菌生理盐水灌胃作为对照。各组小鼠均正常饮食饮水，连续给药2周。每天称量并记录各组小鼠体重。
1.3超声心动图评价小鼠心功能
于给药2周后检测超声心动图，探头频率为30MHz。具体为异氟烷气麻动物后，当小鼠的心率保持在450-500次/分钟时记录M-mode图像。采集胸骨旁长轴切面、心尖四腔切面B-Mode图像。取胸骨旁左室短轴，2D超声示左室短轴切面，在乳头肌水平应用M型超声记录左心室运动情况。功能学指标包括：左室射血分数(LVEF)。比较各组小鼠心脏形态及功能变化。所有测量值均为连续5个心动周期的平均值，由3名经验丰富的技术人员进行。
1.4心肌细胞Western blot检测Wnt通路
分别提取KI组和WT组心肌细胞蛋白，进行Western blot检测。
1.5取材
药物干预2周后处死小鼠，取心脏，比较3组之间心脏大小。
2结果
2.1结果显示：KI组小鼠心功能指标明显低于WT组，KI+Wnt-C59组小鼠心功能改善。(见图1-2)
2.2结果显示：KI组Wnt通路明显激活。(见图3)
2.3结果显示：KI+Wnt-C59组小鼠心脏明显小于KI组。(见图4)
3结论
Wnt信号参与心脏心室重构、心肌肥大、心力衰竭过程。Wnt信号通路的激活与DCM的发生发展相关。基于我们的研究结果，Wnt信号通路抑制剂，Wnt-C59会明显改善衰老联合阿霉素致扩心病小鼠心功能，为Wnt-C59应用于临床治疗扩张型心肌病提供理论依据，同时也为扩心病的治疗提供新的靶点。
Wnt-C59作为新型肿瘤增殖抑制剂，除了对46种肿瘤细胞的增殖产生显著的抑制作用外，Wnt-C59还可以改善半乳糖衰老联合阿霉素致扩心病小鼠心功能，为Wnt-C59应用于临床治疗扩张型心肌病提供理论依据。Wnt-C59是对既往传统的扩心病心肌损伤治疗如强心、利尿、扩血管治疗的重要补充，可以延缓心肌损伤进展，提高患者生活质量。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员，在不脱离本发明原理的前提下，还可以做出若干改进和补充，这些改进和补充也应视为本发明的保护范围。

Claims (6)

  1. Wnt抑制剂在制备治疗SCN5A突变致扩张型心肌病的药物中的应用。
  2. Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用。
  3. Wnt抑制剂Wnt-C59在制备治疗SCN5A-R225Q突变体致扩张型心肌病的药物中的应用。
  4. Wnt抑制剂Wnt-C59在制备改善SCN5A-R225Q突变体致扩张型心肌病患者心脏功能及预后的药物中的应用。
  5. Wnt抑制剂Wnt-C59在制备改善SCN5A-R225Q突变体致扩张型心肌病患者心室腔扩大、心肌收缩功能下降及预后的药物中的应用。
  6. 根据权利要求1-5任一所述的应用，其特征在于，药物剂型选自片剂、粉剂、混悬剂、胶囊剂、丸剂或糖浆。
PCT/CN2021/111105 2020-12-04 2021-08-06 Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用 WO2022116583A1 (zh)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102369187A (zh) * 2009-03-02 2012-03-07 Irm责任有限公司 用作wnt信号调节剂的n-(杂)芳基、2-(杂)芳基取代的乙酰胺类
US10624949B1 (en) * 2015-07-27 2020-04-21 National Technology & Engineering Solutions Of Sandia, Llc Methods for treating diseases related to the wnt pathway
CN112472701A (zh) * 2020-12-04 2021-03-12 复旦大学附属中山医院 Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014078414A1 (en) * 2012-11-13 2014-05-22 Joseph Wu Chemically defined production of cardiomyocytes from pluripotent stem cells
CN109415342A (zh) * 2016-06-22 2019-03-01 诺华股份有限公司 用于治疗纤维化的wnt抑制剂
WO2018045182A1 (en) * 2016-09-01 2018-03-08 The Board Of Regents Of Hte University Of Texas System Disubstituted and trisubtituted 1,2,3-triazoles as wnt inhibitors
JP2021518113A (ja) * 2018-03-16 2021-08-02 ザ ボード オブ トラスティーズ オブ ザ レランド スタンフォード ジュニア ユニバーシティー 心筋細胞を生成及び増殖させるためのｗｎｔアゴニスト、並びに生理活性脂質を用いた試薬及び方法
US20230277540A1 (en) * 2020-06-09 2023-09-07 Genethon Treatment of Dilated Cardiomyopathies

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102369187A (zh) * 2009-03-02 2012-03-07 Irm责任有限公司 用作wnt信号调节剂的n-(杂)芳基、2-(杂)芳基取代的乙酰胺类
US10624949B1 (en) * 2015-07-27 2020-04-21 National Technology & Engineering Solutions Of Sandia, Llc Methods for treating diseases related to the wnt pathway
CN112472701A (zh) * 2020-12-04 2021-03-12 复旦大学附属中山医院 Wnt抑制剂Wnt-C59在制备治疗SCN5A突变致扩张型心肌病的药物中的应用

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
ABRAITYTE AURELIJA, LUNDE IDA G., ASKEVOLD ERIK T., MICHELSEN ANNIKA E., CHRISTENSEN GEIR, AUKRUST PÅL, YNDESTAD ARNE, FIANE ARNT,: "Wnt5a is associated with right ventricular dysfunction and adverse outcome in dilated cardiomyopathy", SCIENTIFIC REPORTS, vol. 7, no. 1, 1 December 2017 (2017-12-01), pages 1 - 10, XP055935755, DOI: 10.1038/s41598-017-03625-9 *
AIJUN SUN,, LEI XU, SHAOCHUN WANG, KAI HU, CHUNYU ZHANG , ZHIYIN YANG , CHENG SHEN , , KEQIANG WANG , SANLI QIAN , ZHENG FAN ,: "PO-309 Altered Cardiac Sodium Channel Function Due to Voltage-Sensor Mutation in Dilated Cardiomyopathy Patients with SCN5A Mutations", PROCEEDINGS OF THE 15TH SCIENTIFIC ANNUAL CONGRESS OF CHINESE SOCIETY OF CARDIOLOGY, 31 August 2013 (2013-08-31), pages 1 - 1, XP055935737 *
DAI XIYAN , HE WENFENG , CAO LINGLING , ZHOU QIONGQIONG , HONG KUI: "Research progress on the relationship between Wnt/β-catenin signaling pathway and arrhythmogenic right ventricular cardiomyopathy", CHINESE JOURNAL OF CARDIAC PACING AND ELECTROPHYSIOLOGY, vol. 26, no. 6, 12 December 2012 (2012-12-12), pages 476 - 477, XP055935754, ISSN: 1007-2659, DOI: 10.13333/j.cnki.cjcpe.2012.06.009 *
DAN LU, DAN BAO, WEI DONG, NING LIU, XU ZHANG, SHAN GAO, WENPING GE, XIANG GAO, LIANFENG ZHANG: "Dkk3 prevents familial dilated cardiomyopathy development through Wnt pathway", LABORATORY INVESTIGATION, vol. 96, no. 2, 1 February 2016 (2016-02-01), pages 239 - 248, XP055525827, ISSN: 0023-6837, DOI: 10.1038/labinvest.2015.145 *
LIU, JINGWEN: "Aberrant Expression Profile of mRNA in Dilated Cardiomyopathy by RNA-Sequence", CHINESE JOURNAL OF HEART FAILURE AND CARDIOMYOPATHY, vol. 2, no. 3, 30 September 2018 (2018-09-30), pages 146 - 150, XP009537017, ISSN: 2096-3076, DOI: 10.3760/cma.j.issn.2096-3076.2018.09.004 *
LU AIZHU, KAMKAR MARYAM, CHU CENCEN, WANG JERRY, GAUDET KAYA, CHEN YAWEN, LIN LAUREN, LIU WEIXIN, MARBÁN EDUARDO, LIANG WENBIN: "Direct and Indirect Suppression of Scn5a Gene Expression Mediates Cardiac Na+ Channel Inhibition by Wnt Signalling", CANADIAN JOURNAL OF CARDIOLOGY, vol. 36, no. 4, 1 April 2020 (2020-04-01), CANADA , pages 564 - 576, XP055935723, ISSN: 0828-282X, DOI: 10.1016/j.cjca.2019.09.019 *
WAN WEI, ZHOU XIAN-SHUN, CAI HONG-ZHUAN: "Current Status of Research in Dilated Cardiomyopathy Pathogenesis in Ion Channels", ADVANCES IN CARDIOVASCULAR DISEASES, vol. 33, no. 4, 5 July 2012 (2012-07-05), pages 551 - 553, XP055935733, ISSN: 1004-3934, DOI: 10.3969/j.issn.1004-3934.2012.04.037 *

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