WO2022266909A1 - 一种增效干细胞治疗心梗的药物及其应用 - Google Patents
一种增效干细胞治疗心梗的药物及其应用 Download PDFInfo
- Publication number
- WO2022266909A1 WO2022266909A1 PCT/CN2021/101922 CN2021101922W WO2022266909A1 WO 2022266909 A1 WO2022266909 A1 WO 2022266909A1 CN 2021101922 W CN2021101922 W CN 2021101922W WO 2022266909 A1 WO2022266909 A1 WO 2022266909A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- myocardial infarction
- stem cells
- glucose
- deoxy
- drug
- Prior art date
Links
- 208000010125 myocardial infarction Diseases 0.000 title claims abstract description 62
- 239000003814 drug Substances 0.000 title claims abstract description 26
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 20
- 229940079593 drug Drugs 0.000 title claims abstract description 17
- 230000002195 synergetic effect Effects 0.000 title claims abstract description 13
- VRYALKFFQXWPIH-PBXRRBTRSA-N (3r,4s,5r)-3,4,5,6-tetrahydroxyhexanal Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)CC=O VRYALKFFQXWPIH-PBXRRBTRSA-N 0.000 claims abstract description 33
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims abstract description 21
- PMMURAAUARKVCB-UHFFFAOYSA-N alpha-D-ara-dHexp Natural products OCC1OC(O)CC(O)C1O PMMURAAUARKVCB-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000011282 treatment Methods 0.000 claims abstract description 13
- 230000004083 survival effect Effects 0.000 claims abstract description 7
- 239000007924 injection Substances 0.000 claims abstract description 5
- 238000002347 injection Methods 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims description 14
- 230000004217 heart function Effects 0.000 claims description 7
- 230000036541 health Effects 0.000 claims description 5
- 208000033774 Ventricular Remodeling Diseases 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 238000009168 stem cell therapy Methods 0.000 claims 1
- 238000009580 stem-cell therapy Methods 0.000 claims 1
- 210000002901 mesenchymal stem cell Anatomy 0.000 abstract description 29
- 230000000694 effects Effects 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 abstract description 4
- 241001465754 Metazoa Species 0.000 abstract description 3
- VRYALKFFQXWPIH-RANCGNPWSA-N (3r,4s,5r)-3,4,5,6-tetrahydroxy-2-tritiohexanal Chemical compound O=CC([3H])[C@@H](O)[C@H](O)[C@H](O)CO VRYALKFFQXWPIH-RANCGNPWSA-N 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 8
- 238000010186 staining Methods 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 6
- 230000006872 improvement Effects 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 5
- 238000002054 transplantation Methods 0.000 description 5
- 239000007928 intraperitoneal injection Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 210000004351 coronary vessel Anatomy 0.000 description 3
- 210000005003 heart tissue Anatomy 0.000 description 3
- 230000000302 ischemic effect Effects 0.000 description 3
- 210000005240 left ventricle Anatomy 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 230000002861 ventricular Effects 0.000 description 3
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 230000000747 cardiac effect Effects 0.000 description 2
- 210000000038 chest Anatomy 0.000 description 2
- 210000001671 embryonic stem cell Anatomy 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 1
- 101100298998 Caenorhabditis elegans pbs-3 gene Proteins 0.000 description 1
- 206010011985 Decubitus ulcer Diseases 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- XQFRJNBWHJMXHO-RRKCRQDMSA-N IDUR Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(I)=C1 XQFRJNBWHJMXHO-RRKCRQDMSA-N 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 210000004504 adult stem cell Anatomy 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000003995 blood forming stem cell Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000004413 cardiac myocyte Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000003205 diastolic effect Effects 0.000 description 1
- 238000002592 echocardiography Methods 0.000 description 1
- 210000002304 esc Anatomy 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 125000002791 glucosyl group Chemical class C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000003601 intercostal effect Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 210000005248 left atrial appendage Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000013227 male C57BL/6J mice Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 238000012141 orotracheal intubation Methods 0.000 description 1
- 210000003540 papillary muscle Anatomy 0.000 description 1
- 210000000062 pectoralis major Anatomy 0.000 description 1
- 210000003516 pericardium Anatomy 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000036387 respiratory rate Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000037390 scarring Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 210000000603 stem cell niche Anatomy 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000012285 ultrasound imaging Methods 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7004—Monosaccharides having only carbon, hydrogen and oxygen atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- 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
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
Definitions
- the invention belongs to the pharmaceutical technology, and relates to a medicine for treating myocardial infarction with synergistic stem cells and its application, specifically the application of 2-deoxy-D-glucose in the preparation of medicine for treating myocardial infarction with synergistic stem cells.
- MI Myocardial infarction
- Ischemic myocardial infarction leads to necrosis and scarring of cardiomyocytes, which in turn affects cardiac function.
- Most of the current drug or device treatments can only relieve the symptoms, but they cannot reverse the damage to the heart tissue.
- heart transplantation can completely improve the state of the heart, it is difficult to be widely used clinically due to factors such as scarcity of donor sources, immune rejection, and expensive treatment costs.
- Stem cells are a series of undifferentiated cells that have the ability to differentiate into specific cell types.
- Stem cells can be divided into embryonic stem cells (Embryonic stem cells, ESCs) and adult stem cells. The latter includes hematopoietic stem cells (Hematopoietic stem cells, HSCs) and mesenchymal stem cells (Mesenchymal stem cells, MSCs).
- Mesenchymal stem cells are present in almost all tissues of the human body with specific stem cell niches.
- Bone marrow-derived mesenchymal stem cells are the first mesenchymal stem cells to be discovered, and they are also the most commonly used stem cells for the treatment of myocardial infarction.
- BM-MSCs Bone marrow-derived mesenchymal stem cells
- 2-Deoxy-D-glucose (2-DG) is a glucose analog that has the effect of interfering with the synthesis of virus-specific glycoproteins, inhibiting the proliferation of herpes monoviruses, RAN and DNA enveloped viruses, and cancer cells; but so far There is no research report on 2-deoxy-D-glucose in myocardial infarction, especially no research on the relationship between 2-DG and stem cells.
- the invention discloses a new application of using 2-deoxy-D-glucose to improve the therapeutic effect of stem cells after myocardial infarction.
- the biggest problem of MSCs treatment of myocardial infarction is the low retention rate and poor survival of transplanted cells in the myocardial infarction site.
- the invention discloses that 2-deoxy-D-glucose can significantly improve the therapeutic effect of mesenchymal stem cells after myocardial infarction.
- the present invention adopts the following technical scheme: the application of 2-deoxy-D-glucose in the preparation of drugs for treating myocardial infarction with synergistic stem cells.
- a drug for synergistic treatment of myocardial infarction the active ingredients of which are 2-deoxy-D-glucose and stem cells. Also disclosed is the application of the medicine for treating myocardial infarction with synergistic effect in the preparation of medicine for treating myocardial infarction.
- 2-DG involved in the present invention in the preparation of drugs or health products for the prevention or treatment of myocardial infarction
- 2-DG can be used to prepare drugs for myocardial infarction protection, health products or food, to achieve 2-deoxy-D-glucose especially
- synergistic stem cells especially mesenchymal stem cells, to improve cardiac function after myocardial infarction, slow down the expansion of ventricles after myocardial infarction, and reduce the size of myocardial infarction.
- the drug prepared by the present invention can be given by intraperitoneal injection or the like, and the dose of 2-deoxy-D-glucose is 300-800 mg/kg/day, such as 500 mg/kg/day.
- MI local intramyocardial transplantation may be superior to intravenous injection or intracoronary transplantation, but regardless of the transplantation route, the ultimate goal is still to optimize the strategy to enhance the survival rate of injected MSCs.
- the effect of the existing mesenchymal stem cell injection in the treatment of myocardial infarction is not good.
- the operation of this method is complicated.
- Myocardial infarction is a cardiovascular disease that seriously endangers human health. With the continuous improvement of people's living standards in our country, the incidence of ischemic myocardial infarction is also increasing.
- the present invention uses 2-deoxy-D-glucose for the first time to improve cardiac function after myocardial infarction, especially in combination with mesenchymal stem cells to increase the effect of stem cells in treating myocardial infarction and reduce the area of myocardial infarction, thereby preventing or treating myocardial infarction.
- FIG 1 shows that 2-DG improves the cardiac function of mice after myocardial infarction treated with MSCs.
- Figure 2 shows that 2-DG improves the ventricular remodeling of mice after myocardial infarction treated with MSCs.
- FIG. 3 shows that 2-DG significantly increased the survival of transplanted MSCs.
- mice C57BL/6J male mice of about 25 g were selected as the experimental subjects, and the myocardial infarction model was established by ligation of the left anterior descending coronary artery. After intraperitoneal injection of anesthesia, orotracheal intubation, connected to an air ventilator, respiratory rate 110 times/min, tidal volume 2.5ml, breath-to-breath ratio 1:1.3.
- the outer skin was incised through the left thoracic longitudinal incision, the pectoralis major muscle was peeled off, the chest was opened through the third and fourth intercostal transverse incisions, the heart was exposed, and the pericardium was torn off with tweezers.
- the left coronary artery can be roughly coursed with the aid of an operating microscope.
- the anterior descending coronary artery (LAD) and a small amount of myocardial tissue were ligated, the depth of the needle was about 1 mm, and the width was controlled within 3 mm; the chest was closed layer by layer.
- the sham operation group only passed under the LAD without knotting, and the rest were the same as the model group.
- the ligation area to the apex of the heart became white with the naked eye.
- the left ventricle tissue was taken for cardiac tissue staining, and obvious fibrosis could be seen, which proved that the myocardial infarction model was successfully established.
- the extracted MSCs were injected at three points at the edge of the myocardial infarction immediately after ligation of the LAD, and the total number of injected cells was 5x10 5 /20 ⁇ l PBS; in the 2-DG group, only 6 hours before the establishment of myocardial infarction and 1 day after myocardial infarction , 2-day after operation, intraperitoneal injection of 2-DG (500 mg/kg/day); MSCs+2-DG group received intraperitoneal injection of 2-DG 6 hours before myocardial infarction, 1 day after myocardial infarction and 2 days after operation DG (500 mg/kg/day), and the rest were the same as those in the MSCs group; mice in the control group (normal saline group) were injected with the same amount of sterile normal saline as a control after ligation of the LAD; Function, detection of myocardial infarction size.
- Example 1 Effective improvement of cardiac function after myocardial infarction Cardiac function after myocardial infarction detected by echocardiography.
- the mice were anesthetized (the same method as before), placed in the left lateral position after hair removal, placed the probe of the cardiac ultrasound diagnostic instrument on the anterior wall of the heart, and took a two-dimensional short-axis view of the left ventricle at the level of the papillary muscle, and recorded M-mode scans at the same time, for 3 consecutive Cardiac cycle measurement of left ventricular ejection fraction (EF), fractional shortening (FS), left ventricular end diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD).
- EF left ventricular ejection fraction
- FS fractional shortening
- LVEDD left ventricular end diastolic diameter
- LVESD left ventricular end systolic diameter
- Example 2 Effective improvement of ventricular remodeling Heart weight Masson staining: mice were sacrificed 28 days after surgery, body weight (BW) and heart weight (heart weight, HW) were measured; HW/BW was calculated. Masson staining: the mice were sacrificed 28 days after the operation, and the left ventricle tissue was taken for heart tissue staining to observe the therapeutic effect. Following the routine Masson staining procedure, observe and take pictures under an ordinary light microscope. The image analysis software Image J was used to analyze the area of each part, and the myocardial infarction area/heart area was calculated. The experimental results are shown in Figure 2, *P ⁇ 0.05, **P ⁇ 0.01.
- Example 3 Increase the survival of mesenchymal stem cells: prepare mesenchymal stem cells stained with CM-DiI, and use the red fluorescence property of CM-DiI to observe the survival rate of transplanted mesenchymal stem cells. Specifically, prepare CM-DiI staining working solution: CM-DiI dye storage solution (Thermo Fisher) with a concentration of 1 mg/mL, take 4 uL of the storage solution and add it to 2 mL of PBS to make the final concentration 2 ug/mL.
- CM-DiI dye storage solution Thermo Fisher
- the heart On the 3rd day after the operation, the heart was taken to prepare a frozen section, and the tissue section of troponin was used for immunofluorescence, the green was marked with troponin, the red was marked with CM-DiI, and the blue was marked with DAPI to mark the nucleus; the heart was digested and prepared on the 3rd day after the operation Cell suspension, the proportion and quantity of CM-DiI positive cells were detected by flow cytometry. The experimental results are shown in Figure 3, *P ⁇ 0.05.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Virology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Vascular Medicine (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一种增效干细胞治疗心梗的药物及其应用，具体为2-脱氧-D-葡萄糖在制备增效干细胞治疗心梗的药物中的应用，动物实验证实，注射2-脱氧-D-葡萄糖后，增加了注射间充质干细胞的存活率，增强间充质干细胞的治疗效果。
Description
本发明属于药物技术，涉及一种增效干细胞治疗心梗的药物及其应用，具体为2-脱氧-D-葡萄糖在制备增效干细胞治疗心梗的药物中的应用。
心肌梗死（MI）是一种严重危害人类健康的心血管疾病，随着我国人民生活水平的不断提高，缺血性心肌梗死的发病率也在不断上升。缺血性心肌梗死会导致心肌细胞坏死和瘢痕形成，进而影响心脏功能。目前药物或器械治疗大多只能缓解症状，但却不能逆转心脏组织损伤。心脏移植虽能彻底改善心脏状态，但因供体来源稀缺、免疫排斥以及昂贵的治疗费用等因素，在临床上很难广泛应用。
干细胞是一系列未分化的细胞，具有分化成特定细胞类型的能力。干细胞按来源可分为胚胎干细胞(Embryonic stem cells,
ESC)和成体干细胞。后者包括造血干细胞(Hematopoietic stem cells,
HSCs)和间充质干细胞(Mesenchymal stem cells,
MSCs)。间充质干细胞几乎存在于人体所有具有特定干细胞龛位的组织中。骨髓来源的间充质干细胞(BM-MSCs)是第一个被发现的间充质干细胞，也是目前最多的用于心梗治疗的干细胞。但是现有技术表明，仅仅依靠干细胞治疗心梗，效果还需改善。
2-脱氧-D-葡萄糖（2-DG）是一种葡萄糖类似物，具有干扰病毒特异性糖蛋白的合成、抑制疱疹单病毒、RAN和DNA包膜病毒、癌症细胞增殖等功效；但迄今为止尚未见2-脱氧-D-葡萄糖在心肌梗死方面的研究报道，尤其未见2-DG与干细胞作用关系的研究。
本发明公开了一种利用2-脱氧-D-葡萄糖改善心梗后干细胞治疗效果的新用途。MSCs治疗心梗的最大问题是移植细胞在心梗部位的滞留率低、存活性差。本发明公开了2-脱氧-D-葡萄糖对心肌梗死后间充质干细胞的治疗效果有明显提升。
本发明采用如下技术方案：2-脱氧-D-葡萄糖在制备增效干细胞治疗心梗的药物中的应用。
2-脱氧-D-葡萄糖在制备治疗心梗的药物中的应用。
2-脱氧-D-葡萄糖和干细胞在制备增效治疗心梗的药物中的应用。
一种增效治疗心梗的药物，其活性成分为2-脱氧-D-葡萄糖和干细胞。还公开了该增效治疗心梗的药物在制备治疗心梗的药物中的应用。
本发明涉及的2-DG在制备预防或治疗心梗药物或保健品中的应用，具体的是2-DG可用于制备心梗保护药物、保健品或者食品，实现2-脱氧-D-葡萄糖尤其是增效干细胞，特别是间充质干细胞改善心梗后心功能、减缓心梗后心室扩张、减小心梗面积的技术效果。
本发明公开的制备的药物，具体的给药方式可采取腹腔注射等，2-脱氧-D-葡萄糖的给药量为300～800mg/kg/天，比如500 mg/kg/天。
考虑到MSCs的移植成功率，MI局部心肌内移植可能比静脉注射或冠状动脉内移植更为优越，但无论采用何种移植途径，最终目的仍然是优化策略，以增强注射的MSCs的存活率。现有技术下，由于MI导致心脏部位环境的复杂，使得现有间充质干细胞注射治疗心梗的效果欠佳，研究者采用各种方法以提高其对心梗的治疗效果，比如体外采用细胞因子培养间充质干细胞后回输等，此方法操作复杂。心肌梗死是一种严重危害人类健康的心血管疾病，随着我国人民生活水平的不断提高，缺血性心肌梗死的发病率也在不断上升。本发明首次采用2-脱氧-D-葡萄糖改善心梗后心功能，尤其是与间充质干细胞结合，增加干细胞治疗心梗的效果，减小心梗面积，从而可以预防或者治疗心梗。
图1为2-DG改善MSCs治疗心梗后小鼠的心功能。
图2为2-DG改善MSCs治疗心梗后小鼠的心室重构。
图3为2-DG明显增加移植MSCs存活。
以下所列实施例，仅为帮助本领域技术人员更全面理解本发明，但不以任何方式限制本发明。本发明涉及的原料试剂以及建模、测试都是本领域常规技术。
所使用的主要材料和来源分别如下：C57BL/6J小鼠（昭衍（苏州）新药研究中心提供，此实验由苏州大学伦理委员会批准）；小动物呼吸机（上海奥尔科特生物科技有限公司，上海）；手术器械（六六视觉公司，苏州）；缝合针线（上海浦东金环医疗用品有限公司，上海）；小动物超声影像系统（Visual Sonics Vevo 2100）；间充质干细胞（MSCs），采用现有技术从8周雄性C57BL/6J小鼠股骨骨髓中提取，常规培养第六代以后用于实验；2-脱氧-D-葡萄糖（2-DG，苏州天可贸易有限公司，苏州），溶于生理盐水（浓度为31.25mg/mL），备用。
小鼠心肌梗死模型的建立：选用25g左右的C57BL/6J雄性小鼠为实验对象，采用左冠状动脉前降支结扎法制作心梗模型。腹腔注射麻醉后，经口气管插管，接空气呼吸机，呼吸频率110次/min，潮气量2.5ml，吸呼比1:1.3。右侧卧位，左胸纵切口切开外层皮肤，剥离胸大肌，第三、四肋间横切口开胸，暴露心脏，用镊子撕开心包膜。借助手术显微镜可见左冠状动脉大致走行。在左心耳下缘约1.5mm处，将冠状动脉前降支（LAD） 连同少量心肌组织一起结扎，进针深度约1 mm，宽度控制在3 mm以内；逐层关胸。
假手术组仅穿过LAD下方不打结，其余同模型组。
结扎后肉眼可见结扎处至心尖变白，1 周后取左心室组织进行心脏组织染色，可看到明显的纤维化，证明心梗模型建立成功。
MSCs组在结扎LAD后当即在心梗边缘区分三点注射提取的MSCs，注射细胞总数量为5x10
5/20 μl PBS；2-DG组仅在心梗建立前6h、心梗术后第1天、术后第2天腹腔注射2-DG（500 mg/kg/天）；MSCs+2-DG组在心梗建立前6h、心梗术后第1天、术后第2天腹腔注射2-DG（500 mg/kg/天），其余同MSCs组；对照组（生理盐水组）小鼠在结扎LAD后注射等量无菌生理盐水作为对照；三组喂养方式一致，然后进行心梗后心功能、心梗面积检测。
实施例1 有效改善心梗后心功能: 心脏超声检测心梗后心功能。小鼠麻醉（方法同前），脱毛后左侧卧位，将心脏超声诊断仪探头置于心前壁，于乳头肌水平取左室二维短轴观，同时记录M型扫描，连续3个心动周期测量左室射血分数（EF）、缩短分数（FS）和左室舒张末径（LVEDD）和左室收缩末径（LVESD）。实验结果见图1，*
P < 0.05；#
P < 0.05。
实施例2 有效改善心室重构: 心脏重量Masson染色：术后28天处死小鼠，测量小鼠体重（body weight，BW）、心脏重量（heart weight，HW）；计算HW/BW。Masson染色：术后28天处死小鼠，取左心室组织进行心脏组织染色，观察治疗效果。按常规Masson染色步骤进行，在普通光学显微镜下观察并拍照。采用图像分析软件Image J分析各部分面积，计算心梗面积/心脏面积。实验结果见图2，*P < 0.05，**P < 0.01。
实施例3 增加间充质干细胞存活: 制备CM-DiI染色的间充质干细胞，利用CM-DiI的红色荧光特性观察移植间充质干细胞的存活率。具体的，配置CM-DiI染色工作液：CM-DiI染料储存液（赛默飞世尔）浓度为1mg/mL，取4uL储存液加到2mL PBS中，使终浓度为2ug/mL。取拟注射的MSCs（500000个），常规离心后用CM-DiL染色工作液重悬，37℃下孵育5min，再放入 4℃下放置 15 分钟，然后用PBS彻底清洗3遍，加入20uL PBS重悬用于心梗后心肌内注射。
术后第3天取心脏制备冰冻切片，做肌钙蛋白的组织切片免疫荧光，绿色标记肌钙蛋白，红色标记CM-DiI，蓝色表示DAPI标记细胞核；术后第3天取心脏消化制备单细胞悬液，流式检测CM-DiI阳性细胞比例和数量。实验结果见图3，*P<0.05。
【结论】上述实验的结果综合证明：2-DG能有效提高MSCs改善心梗的功能。
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来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (10)
- 一种增效治疗心梗的药物，其特征在于，所述药物的活性成分为2-脱氧-D-葡萄糖和干细胞。
- 根据权利要求1所述增效治疗心梗的药物，其特征在于，所述药物为注射制剂。
- 2-脱氧-D-葡萄糖在制备增效干细胞治疗心梗的药物中的应用。
- 根据权利要求3所述的应用，其特征在于，所述药物为注射制剂。
- 根据权利要求3所述的应用，其特征在于，2-脱氧-D-葡萄糖改善心梗后干细胞治疗的心功能与心室重构。
- 2-脱氧-D-葡萄糖在制备治疗心梗的药物中的应用。
- 2-脱氧-D-葡萄糖和干细胞在制备增效治疗心梗的药物中的应用。
- 2-脱氧-D-葡萄糖在制备预防心梗的保健品中的应用。
- 2-脱氧-D-葡萄糖在制备增加干细胞在心梗部位存活率的药物中的应用。
- 权利要求1所述增效治疗心梗的药物在制备治疗心梗的药物中的应用。
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2021/101922 WO2022266909A1 (zh) | 2021-06-23 | 2021-06-23 | 一种增效干细胞治疗心梗的药物及其应用 |
| US18/035,685 US20240108658A1 (en) | 2021-06-23 | 2021-06-23 | Synergistic stem cell-based medication for treating myocardial infarction and related application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2021/101922 WO2022266909A1 (zh) | 2021-06-23 | 2021-06-23 | 一种增效干细胞治疗心梗的药物及其应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2022266909A1 true WO2022266909A1 (zh) | 2022-12-29 |
Family
ID=84545062
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2021/101922 WO2022266909A1 (zh) | 2021-06-23 | 2021-06-23 | 一种增效干细胞治疗心梗的药物及其应用 |
Country Status (2)
| Country | Link |
|---|---|
| US (1) | US20240108658A1 (zh) |
| WO (1) | WO2022266909A1 (zh) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103800346A (zh) * | 2014-01-28 | 2014-05-21 | 四川省医学科学院（四川省人民医院） | 葡萄糖类似物在制备治疗或预防细胞死亡疾病药物中的应用 |
| WO2018088693A1 (ko) * | 2016-11-11 | 2018-05-17 | 사회복지법인 삼성생명공익재단 | 2ｄｇ를 이용한 줄기세포 역량 향상 방법 |
-
2021
- 2021-06-23 US US18/035,685 patent/US20240108658A1/en active Pending
- 2021-06-23 WO PCT/CN2021/101922 patent/WO2022266909A1/zh active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103800346A (zh) * | 2014-01-28 | 2014-05-21 | 四川省医学科学院（四川省人民医院） | 葡萄糖类似物在制备治疗或预防细胞死亡疾病药物中的应用 |
| WO2018088693A1 (ko) * | 2016-11-11 | 2018-05-17 | 사회복지법인 삼성생명공익재단 | 2ｄｇ를 이용한 줄기세포 역량 향상 방법 |
Non-Patent Citations (4)
| Title |
|---|
| HU XIN-YU, GAO ZHENG-LIANG; XU JUN: "2-DG inhibits proliferation and promotes astrocytic differentiation of neural stem cells", JOURNAL OF TONGJI UNIVERSITY (MEDICAL SCIENCE), vol. 37, no. 6, 31 December 2016 (2016-12-31), pages 35 - 40, XP093017264, ISSN: 1008-0392, DOI: 10.16118/j.1008-0392.2016.06.007 * |
| LI LANG, DONG LIANG, HUI JIAOJIE GOOFY, WANG QIUHUI, YANG LAN,ZHANG JIANGQIAN, YAN JIE: "Effects of Hippo signaling on anti-oxidative stress of mesenchymal stem cells in vitro", CHINESE CRITICAL CARE MEDICINE - ZHONGHUA WEIZHONGBING JIJIU YIXUE, ZHONGHUA YIXUEHUI, CN, vol. 30, no. 4, 10 April 2018 (2018-04-10), CN , pages 296 - 301, XP009542104, ISSN: 2095-4352, DOI: 10.3760/cma.j.issn.2095-4352.2018.04.002 * |
| TANI M., SHINMURA K., EBIHARA Y., ASAKURA Y.: "Inhibition of glycolysis or increased perfusate H+ buffering capacity, but not their combination, attenuates myocardial stunning", CARDIOVASCULAR RESEARCH, OXFORD UNIVERSITY PRESS, GB, vol. 27, no. 9, 1 September 1993 (1993-09-01), GB , pages 1645 - 1650, XP009542091, ISSN: 0008-6363, DOI: 10.1093/cvr/27.9.1645 * |
| TENG XIAOMEI, CHEN LEI, CHEN WEIQIAN, YANG JUNJIE, YANG ZIYING, SHEN ZHENYA: "Mesenchymal Stem Cell-Derived Exosomes Improve the Microenvironment of Infarcted Myocardium Contributing to Angiogenesis and Anti-Inflammation", CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, KARGER BASEL, CH, vol. 37, no. 6, 1 January 2015 (2015-01-01), CH , pages 2415 - 2424, XP093017261, ISSN: 1015-8987, DOI: 10.1159/000438594 * |
Also Published As
| Publication number | Publication date |
|---|---|
| US20240108658A1 (en) | 2024-04-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20070042016A1 (en) | Methods and Systems for Treating Injured Cardiac Tissue | |
| US20140106447A1 (en) | Compositions and methods for recruiting stem cells | |
| US20070093748A1 (en) | Methods and systems for treating injured cardiac tissue | |
| CN104922059A (zh) | 一种脐带间充质干细胞注射液及其制备方法和应用 | |
| CN105395571A (zh) | 间充质干细胞分泌的外泌小体治疗心肌梗死的新用途 | |
| Mehta | Rhabdomyoma and ventricular preexcitation syndrome: A report of two cases and review of literature | |
| EP2393916A2 (en) | Therapeutic use of specialized endothelial progenitor cells | |
| CN108567775B (zh) | 硫辛酸在制备治疗压力负荷性心肌损伤的药物组合物中的应用 | |
| WO2023185085A1 (zh) | Pd1抑制剂在制备心脏成纤维细胞转分化抑制剂中的用途 | |
| WO2022266909A1 (zh) | 一种增效干细胞治疗心梗的药物及其应用 | |
| CN113425731B (zh) | 一种增效干细胞治疗心梗的药物及其应用 | |
| Law et al. | Myoblast therapies constitute a safe and efficacious platform technology of regenerative medicine for the human health industry | |
| WO2019153364A1 (zh) | 一种抗缺氧损伤的干细胞制剂及其制备方法与在制备治疗急性心梗药物中的应用 | |
| CN113855650B (zh) | 一种免疫代谢心梗贴片及其制备方法与应用 | |
| CN107625781B (zh) | miRNA抑制子在制备防治心肌梗死药物中的应用 | |
| WO2023044689A1 (zh) | 一种免疫代谢心梗贴片及其制备方法与应用 | |
| US10828338B2 (en) | Methods for improving cognition and slowing cognitive impairment using nonviable lyophilized pluripotent stem cells | |
| Olivares et al. | Cellular cardiomyoplasty in large myocardial infarction: Can the beneficial effect be enhanced by ACE‐inhibitor therapy? | |
| CN111345917A (zh) | 小鼠心肌梗死后的心肌注射造模方法及装置 | |
| WO2023023995A1 (zh) | 一种防治心肌缺血后心功能衰竭的药物及其应用 | |
| CN115998737B (zh) | 阿莫地喹在制备治疗压力负荷性心肌损伤的药物中的应用 | |
| CN111166757A (zh) | 马钱苷促进血管新生治疗心肌缺血的用途 | |
| CN110664794A (zh) | 氨基氧乙酸在制备预防或治疗心梗的药物中的应用 | |
| Polak et al. | Suspicion of myocarditis in a patient with mitral valve prolapse | |
| Carraro et al. | Cardiac-bio-assists: biological approaches to support or repair cardiac muscle |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 21946407 Country of ref document: EP Kind code of ref document: A1 |
|
| WWE | Wipo information: entry into national phase |
Ref document number: 18035685 Country of ref document: US |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 21946407 Country of ref document: EP Kind code of ref document: A1 |