CN109364055B - 丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用 - Google Patents
丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用 Download PDFInfo
- Publication number
- CN109364055B CN109364055B CN201811286053.9A CN201811286053A CN109364055B CN 109364055 B CN109364055 B CN 109364055B CN 201811286053 A CN201811286053 A CN 201811286053A CN 109364055 B CN109364055 B CN 109364055B
- Authority
- CN
- China
- Prior art keywords
- danshensu
- lupus nephritis
- treating
- medicine
- application
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- PAFLSMZLRSPALU-MRVPVSSYSA-N (2R)-3-(3,4-dihydroxyphenyl)lactic acid Chemical compound OC(=O)[C@H](O)CC1=CC=C(O)C(O)=C1 PAFLSMZLRSPALU-MRVPVSSYSA-N 0.000 title claims abstract description 53
- PAFLSMZLRSPALU-UHFFFAOYSA-N Salvianic acid A Natural products OC(=O)C(O)CC1=CC=C(O)C(O)=C1 PAFLSMZLRSPALU-UHFFFAOYSA-N 0.000 title claims abstract description 52
- PAFLSMZLRSPALU-QMMMGPOBSA-N Danshensu Natural products OC(=O)[C@@H](O)CC1=CC=C(O)C(O)=C1 PAFLSMZLRSPALU-QMMMGPOBSA-N 0.000 title claims abstract description 51
- 208000005777 Lupus Nephritis Diseases 0.000 title claims abstract description 31
- 239000003814 drug Substances 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title description 4
- 102000015694 estrogen receptors Human genes 0.000 claims abstract description 8
- 108010038795 estrogen receptors Proteins 0.000 claims abstract description 8
- 230000000694 effects Effects 0.000 claims description 15
- 239000007935 oral tablet Substances 0.000 claims description 9
- 229940096978 oral tablet Drugs 0.000 claims description 9
- 229940102550 Estrogen receptor antagonist Drugs 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 230000019491 signal transduction Effects 0.000 claims description 3
- 239000013543 active substance Substances 0.000 claims 1
- 241000699670 Mus sp. Species 0.000 abstract description 34
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 abstract description 26
- 210000003734 kidney Anatomy 0.000 abstract description 17
- 210000004369 blood Anatomy 0.000 abstract description 15
- 239000008280 blood Substances 0.000 abstract description 15
- 229940109239 creatinine Drugs 0.000 abstract description 13
- 230000004083 survival effect Effects 0.000 abstract description 13
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 abstract description 9
- 230000008021 deposition Effects 0.000 abstract description 8
- 238000002474 experimental method Methods 0.000 abstract description 8
- 229940079593 drug Drugs 0.000 abstract description 7
- 201000008383 nephritis Diseases 0.000 abstract description 4
- 230000001225 therapeutic effect Effects 0.000 abstract description 4
- 108010002350 Interleukin-2 Proteins 0.000 abstract description 2
- 210000001744 T-lymphocyte Anatomy 0.000 abstract description 2
- 230000004069 differentiation Effects 0.000 abstract description 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 230000008685 targeting Effects 0.000 abstract description 2
- 229940126585 therapeutic drug Drugs 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 21
- 229930189533 tanshinol Natural products 0.000 description 21
- 241000699666 Mus <mouse, genus> Species 0.000 description 12
- 239000000243 solution Substances 0.000 description 10
- LRJOMUJRLNCICJ-JZYPGELDSA-N Prednisolone acetate Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(=O)COC(=O)C)(O)[C@@]1(C)C[C@@H]2O LRJOMUJRLNCICJ-JZYPGELDSA-N 0.000 description 9
- 238000000034 method Methods 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 210000002700 urine Anatomy 0.000 description 8
- 108060001084 Luciferase Proteins 0.000 description 7
- 239000005089 Luciferase Substances 0.000 description 7
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 6
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 6
- 229960005309 estradiol Drugs 0.000 description 6
- 229930182833 estradiol Natural products 0.000 description 6
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 6
- 229960002800 prednisolone acetate Drugs 0.000 description 5
- 239000002904 solvent Chemical group 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 4
- 238000003125 immunofluorescent labeling Methods 0.000 description 4
- 230000003907 kidney function Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 238000001890 transfection Methods 0.000 description 4
- 239000012096 transfection reagent Substances 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 229940122880 Estrogen receptor agonist Drugs 0.000 description 3
- 108700008625 Reporter Genes Proteins 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 201000001474 proteinuria Diseases 0.000 description 3
- 208000031648 Body Weight Changes Diseases 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 206010061481 Renal injury Diseases 0.000 description 2
- 230000004579 body weight change Effects 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 229960004397 cyclophosphamide Drugs 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000003862 glucocorticoid Substances 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 229960003444 immunosuppressant agent Drugs 0.000 description 2
- 230000001861 immunosuppressant effect Effects 0.000 description 2
- 239000003018 immunosuppressive agent Substances 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 208000037806 kidney injury Diseases 0.000 description 2
- 238000003670 luciferase enzyme activity assay Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000000902 placebo Substances 0.000 description 2
- 229940068196 placebo Drugs 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 229940122739 Calcineurin inhibitor Drugs 0.000 description 1
- 101710192106 Calcineurin-binding protein cabin-1 Proteins 0.000 description 1
- 102100024123 Calcineurin-binding protein cabin-1 Human genes 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 206010018364 Glomerulonephritis Diseases 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 206010035148 Plague Diseases 0.000 description 1
- 208000002500 Primary Ovarian Insufficiency Diseases 0.000 description 1
- 208000001647 Renal Insufficiency Diseases 0.000 description 1
- 206010062237 Renal impairment Diseases 0.000 description 1
- 206010038536 Renal tubular atrophy Diseases 0.000 description 1
- 108091027981 Response element Proteins 0.000 description 1
- 208000034189 Sclerosis Diseases 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 210000001736 capillary Anatomy 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000020832 chronic kidney disease Diseases 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000005094 computer simulation Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000028208 end stage renal disease Diseases 0.000 description 1
- 201000000523 end stage renal failure Diseases 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 210000005086 glomerual capillary Anatomy 0.000 description 1
- 210000000585 glomerular basement membrane Anatomy 0.000 description 1
- 230000024924 glomerular filtration Effects 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 201000006370 kidney failure Diseases 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000001325 log-rank test Methods 0.000 description 1
- 206010025135 lupus erythematosus Diseases 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 210000003584 mesangial cell Anatomy 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical group C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000003032 molecular docking Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- RTGDFNSFWBGLEC-SYZQJQIISA-N mycophenolate mofetil Chemical compound COC1=C(C)C=2COC(=O)C=2C(O)=C1C\C=C(/C)CCC(=O)OCCN1CCOCC1 RTGDFNSFWBGLEC-SYZQJQIISA-N 0.000 description 1
- 229960004866 mycophenolate mofetil Drugs 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 231100000857 poor renal function Toxicity 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 206010036601 premature menopause Diseases 0.000 description 1
- 208000017942 premature ovarian failure 1 Diseases 0.000 description 1
- 230000009103 reabsorption Effects 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 229960002385 streptomycin sulfate Drugs 0.000 description 1
- 210000005239 tubule Anatomy 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
Images
Classifications
-
- 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
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
Landscapes
- Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Transplantation (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Epidemiology (AREA)
- Urology & Nephrology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
本发明提供了一种丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用，属于肾炎药物技术领域。丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用。丹参素可通过抑制雌激素受体信号通路，竞争性地与ERα结合，降低IL‑2的产生，调节T细胞分化发挥治疗狼疮性肾炎的作用，说明治疗药物具有靶向新型、安全性高的特点。实验证明：丹参素能够提高MRL/lpr小鼠的生存率，降低血肌酐和血尿素氮水平，减少肾脏免疫复合物沉积，证实丹参素对于狼疮性肾炎具有良好的治疗作用。
Description
技术领域
本发明属于肾炎药物技术领域，具体涉及一种丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用。
背景技术
系统性红斑狼疮(systemic lupus erythematosus，SLE)是一种严重的可累及全身多脏器的自身免疫性疾病，病情呈反复发作与缓解交替进行。其中，肾脏是SLE发病过程中最常见的被累及器官。据临床统计，狼疮患者的肾炎发病率高达50％～70％，狼疮性肾炎(lupus nephritis，LN)是系统性红斑狼疮累及肾脏所引起的一种免疫复合物性肾炎，多为肾小球性肾炎，但也经常伴随着肾小管、肾间质和毛细血管的损伤。其病理变化主要有系膜细胞增生，系膜基质增多，在肾小球、肾间质均常见伴有明显的淋巴细胞和巨噬细胞的浸润；也有新月体纤维化，肾小球毛细血管内增生，肾间质硬化，肾小管萎缩等现象出现，是造成SLE患者死亡的主要原因。
狼疮性肾炎是SLE最严重的并发症之一，可加重肾脏负担，并且进展快速，病死率较高，狼疮性肾炎病人中70％右左都会出现肾脏损伤，10％～15％最终发展为终末期肾衰竭，目前尚无完全治愈方法，因此，如何控制和延缓疾病的进展，提高患者的存活率，是临床面临的巨大挑战。
目前，临床上狼疮性肾炎的治疗常需要大剂量糖皮质激素联合免疫抑制剂、细胞毒药物，如环磷酰胺、吗替麦考酚酯、钙调神经磷酸酶抑制剂等，容易降低患者抵抗力，并发感染。环磷酰胺还会导致卵巢早衰，增加肿瘤风险。另外，上述治疗仅对80％的LN患者有效，其中55％-80％的患者在长达12-24个月才能起效，10％-20％仍然无效，10％-20％患者在10年内发展为终末期肾衰，50％患者会经历复发。为减少复发，在长期服用小剂量糖皮质激素和免疫抑制剂的过程中，也会产生一系列不良反应，如抑制自身免疫，容易诱发或者加重感染，抑制生殖系统功能，增加肿瘤发生，破坏造血功能，导致正常的机体免疫功能受损等。因此，寻找具有疗效显著且安全性高的新靶点药物成为目前狼疮肾炎治疗亟待解决的问题。
发明内容
有鉴于此，本发明的目的在于提供丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用，丹参素具有疗效显著且安全性高的特点。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了一种具有式(1)所示结构的丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用；
优选的，所述丹参素通过作用雌激素受体信号通路实现治疗狼疮性肾炎的作用。
优选的，所述丹参素作为雌激素受体拮抗剂竞争性地与ERα结合，抑制ERα的表达。
优选的，所述药物包括口服片剂；所述口服片剂还包括医药领域可接受的辅料。
优选的，所述丹参素的有效剂量为90.9～540.9mg/kg。
本发明提供的具有式(1)所示结构的丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用。丹参素可通过抑制雌激素受体信号通路，竞争性地与ERα结合，降低IL-2的产生，调节T细胞分化发挥治疗狼疮性肾炎的作用，说明治疗药物具有靶向新型、安全性高的特点。实验证明：丹参素能够提高MRL/lpr小鼠的生存率，降低血肌酐和血尿素氮水平，减少肾脏免疫复合物沉积，证实丹参素对于狼疮性肾炎具有良好的治疗作用。
附图说明
图1为丹参素显著提高MRL/lpr小鼠生存率；
图2为丹参素显著降低MRL/lpr小鼠蛋白尿水平；
图3为丹参素改善MRL/lpr小鼠肾功能；图3-A为血肌酐(CRE)水平；图3-B为MRL/lpr小鼠血尿素氮(BUN)水平；**p<0.01，***p<0.001丹参素给药组vs模型组；
图4为丹参素减少MRL/lpr小鼠肾脏免疫复合物沉积；图4-A为肾脏冰冻切片Ig G免疫荧光染色代表图片；图4-B为40倍视野下IgG免疫荧光累计光密度(IntegratedOptical Density，IOD)统计，*p<0.05，丹参素给药组vs模型组；
图5为丹参素、雌二醇与ERα的结合模式图；图5-A为丹参素与ERα的结合；图5-B为雌二醇与ERα的结合；
图6为丹参素与ERα的结合能力作用，与空白对照组组比较，**表示P<0.01，***P<0.001；
图7为转染ERαHEK-293细胞蛋白表达的结果，与空白对照组组比较，***P<0.001；与E2组比较，###表示P<0.001。
具体实施方式
本发明提供了一种具有式(1)所示结构的丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用；
在本申请中，所述丹参素是通过作用雌激素受体信号通路实现治疗狼疮性肾炎的作用。所述丹参素优选作为雌激素受体拮抗剂竞争性地与ERα结合，抑制ERα的表达。
在本申请中，所述药物优选包括口服片剂；所述口服片剂还包括医药领域可接受的辅料。所述丹参素的有效浓度优选为90.9～540.9mg/kg。本发明对所述辅料的种类没有特殊限制，采用本领域技术人员所熟知的口服片剂的辅料即可。
本发明对所述口服片剂的制备方法没有特殊限制，采用本领域技术人员所熟知的口服片剂制备方法即可。
下面结合实施例对本发明提供的一种丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用进行详细的说明，但是不能把它们理解为对本发明保护范围的限定。
实施例1
采用8周龄雌性MRL/lpr小鼠，每天一次口服给予60mg/kg的丹参素至小鼠22周龄，以给药阳性药醋酸泼尼松龙(PNS)组和溶剂组(Vehicle)为对照。给药期间每周监测体重变化，并记录小鼠生存状况。至小鼠22周龄停止给药，观察丹参素在MRL/lpr小鼠上的治疗效果，给药期间监测小鼠治疗期间生存状况，试验终点时统计小鼠累计生存率，并采用log-rank test进行统计学分析。
结果见图1。由图1可知，丹参素组小鼠生产25周，存活率达到100％，而三七总皂苷组小鼠在15周时，存活率为80％，而溶剂对照组从第10周开始小鼠出现死亡，到25周时，小鼠存活率达到50％。这表明丹参素显著提高MRL/lpr小鼠生存率。同时通过该实例可以证实丹参素对狼疮性肾炎的治疗效果比目前临床上治疗常用的激素药物醋酸泼尼松龙具有优势，效果好，安全性高。
实施例2
采用8周龄雌性MRL/lpr小鼠，每天一次口服给予60mg/kg的丹参素至小鼠22周龄，以给药阳性药醋酸泼尼松龙(PNS)组和溶剂组(Vehicle)为对照。给药期间每周监测蛋白尿，体重变化。至小鼠22周龄停止给药。采用考马斯亮蓝方法检测小鼠尿液中蛋白浓度，将小鼠尿液用PBS稀释，用10μl尿液样品与100μl考马斯亮蓝反应液室温反应15min，595nm处读取OD值。使用牛血清白蛋白(BSA)做标准曲线，测得的值乘以样品稀释倍数即为尿蛋白浓度。狼疮性肾炎发病过程中，肾脏发生损伤，导致肾小球滤过功能和重吸收能力下降，尿中蛋白浓度升高。随着MRL/lpr小鼠狼疮性肾炎的发展，小鼠尿蛋白水平逐渐升高。考马斯亮蓝法检测小鼠尿液中蛋白含量的结果见图2。
图2结果显示模型组小鼠尿蛋白浓度随着周龄的增加而逐渐升高，而丹参素治疗能够显著降低小鼠蛋白尿浓度。
实施例3
采用8周龄雌性MRL/lpr小鼠，每天一次口服给予60mg/kg的丹参素至小鼠22周龄，以给药阳性药醋酸泼尼松龙(PNS)组和溶剂组(Vehicle)为对照。观察丹参素在MRL/lpr小鼠上的治疗效果。尿素氮和肌酐分别是人体蛋白质和肌肉的代谢产物，肾脏是其终排泄器官。肾功能受损后，血中尿素氮和肌酐无法有效排除，血尿素氮(BUN)和血肌酐(CRE)的浓度因潴留而迅速升高。因此血清中BUN和CRE水平是临床上反映肾功能的主要指标。
结果如图3所示，图3为丹参素改善MRL/lpr小鼠肾功能；其中图3-A为血肌酐(CRE)水平；图3-B为MRL/lpr小鼠血尿素氮(BUN)水平；**p<0.01，***p<0.001丹参素给药组vs模型组。由图3结构可知，丹参素能够有效地降低MRL/lpr小鼠血尿素氮以及血肌酐水平，改善小鼠肾功能。
实施例4
丹参素减少MRL/lpr小鼠肾脏免疫复合物沉积
采用8周龄雌性MRL/lpr小鼠，每天一次口服给予60mg/kg的丹参素至小鼠22周龄，以给药阳性药醋酸泼尼松龙(PNS)组和溶剂组(Vehicle)为对照。制备小鼠肾脏冰冻切片进行IgG免疫荧光染色。使用Image-ProPlus6.0软件分析，计算荧光强度，面积和累计光密度，检测小鼠肾脏免疫复合物沉积。狼疮性肾炎发病时，B细胞分泌大量自身抗体，与自身抗原结合后形成免疫复合物，沉积在肾小球基底膜，造成肾损伤。对小鼠肾脏冰冻切片进行Ig G免疫荧光染色的结果见图4。
图4为丹参素减少MRL/lpr小鼠肾脏免疫复合物沉积。图4-A为肾脏冰冻切片Ig G免疫荧光染色代表图片；图4-B为40倍视野下IgG免疫荧光累计光密度(IntegratedOptical Density，IOD)统计，其中*p<0.05，丹参素给药组vs模型组。由图4可知，丹参素能够显著减少小鼠肾脏免疫复合物沉积。
实施例5
计算机模拟分子对接丹参素和雌激素受体结合实验表明，丹参素与雌二醇具有相同的结合位点，且丹参素与ERα的结合能为-8.2kcal/mol(图5-A)，雌二醇的结合能为-5.4kcal/mol(图5-B)。
采用不同浓度的丹参素采用ERαbinding试剂盒(购自life公司)检测丹参素与ERα结合能力，结果图6所示。
图6为丹参素与ERα的结合能力作用，其中空白对照组比较，**表示P<0.01，***P<0.001。这表明丹参素更易与雌激素受体结合。由图6可知，丹参素存在竞争性地与ERα结合的作用，且在剂量范围内，随着浓度增大，结合作用增强。
实施例6
1.转染ERα-ERE荧光素酶报告基因的HEK-293细胞的luciferase实验
分别构建ERα表达质粒(该质粒可找专业的基因公司构建)与雌激素反应元件ERE的荧光素酶报告基因质粒(2ERE-39PRL-Luc)(该质粒有商品化的可购买使用)转染至HEK-293细胞。具体细胞培养和转染的方法如下：HEK-293细胞培养在温度为37℃，二氧化碳浓度为5％的细胞培养箱(Sanyo)中。使用培养基为含有10％灭活的胎牛血清的DMEM培养基。配置1L培养基：DMEM粉末培养基(Dulbecco’s Modified Eagle Medium，Invitrogen)，3.7gNaHCO3，硫酸链霉素和氨苄青霉素各0.1g，用1mol/L HCl溶液调pH值至7.2～7.6之间。使用滤膜过滤法除去细菌。
HEK-293细胞为贴壁生长型，当细胞密度达到80％～90％进行细胞传代。传代时，使用胰酶消化细胞，37℃消化1min吸取胰酶，加入DMEM培养液轻柔吹打细胞直至细胞细胞份数均匀，团块消失，随后根据实验需要移取一定体积的细胞悬浮液至新鲜的完全培养基中，细胞培养36h后，使用高效真核转染试剂VigoFect转染HEK-293细胞，转染前1h，细胞密度达到40～60％为宜，并更换新鲜的完全培养基。转染试剂的用量为0.45μL转染试剂/1μLDNA，使用相同体积的生理盐水与转染试剂充分混合，室温防治5min以上，随后将需要转染的DNA样品和Mixer充分混合，室温防治至少15min以后，将混合液体滴在细胞培养液中即可。如果转染时细胞密度低于30％，转染后4～6h更换培养液。
2、丹参素与雌激素受体激动剂雌二醇E2同时给药的实验
稳定转染的ERα-ERE荧光素酶报告基因的HEK-29细胞正常培养于添加10％FBS和1％PS的DMEM高糖培养基中，置于37℃、5％CO2饱和湿度培养箱内培养。选择对数生长期细胞，胰酶消化后，添加无酚红培养基吹散细胞，以80000个/孔的浓度接种于96孔板内，180μl/孔。培养24h待细胞贴壁后，丹参素各剂量组(10-2,10-3,10-4,10-5,10-6M)加入对应浓度的丹参素溶液20μl，ICI组加10-6M的雌激素受体拮抗剂ICI溶液20μl，E2组加10-7M的雌激素受体激动剂E2溶液20μl，E2+丹参素各剂量组先吸出20μl培养基再同时加对应的E2和丹参素溶液20μl，空白对照组加入20μl的DMEM无酚红培养基，每组设3复孔。培养24h后，移去培养基，每孔加入50μl 1×lysis buffer，低速振摇15min取20μl转移至白色微孔板，每孔加入50μl Luciferase assay试剂，立即测定Luciferase活性。
结果如图7所示。其中图7为转染ERα-ERE荧光素酶报告基因的HEK-293细胞蛋白表达的结果。与空白对照组组比较，***P<0.001；与E2组比较，###表示P<0.001。
通过转染ERα-ERE荧光素酶报告基因的HEK-293细胞的luciferase实验表明，与正常组相比，丹参素在1～100nmol/L浓度范围内均能显著抑制ERα的表达，作用效果与雌激素受体拮抗剂ICI一致(图7-A)。
通过丹参素与雌激素受体激动剂雌二醇E2同时给药的实验表明了相同的结果，丹参素在10～100nmol/L浓度范围内均能显著抑制E2的促进表达作用，作用效果与ICI一致，显示丹参素具有雌激素受体拮抗剂样作用(图7-B)。
由上述实施例可知，与模型组相比：丹参素给药组小鼠在25周内存活率为100％，丹参素给药组小鼠蛋白尿显著下降，生存率显著升高，血肌酐和血尿素氮水平也明显降低，肾脏疫复合物沉积减少，初步证实丹参素对于狼疮性肾炎具有良好的治疗作用。
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 (5)
1.一种具有式(1)所示结构的丹参素作为唯一活性物质在制备治疗和/或预防狼疮性肾炎的药物中的应用；
2.根据权利要求1所述的应用，其特征在于，所述丹参素通过作用雌激素受体信号通路实现治疗狼疮性肾炎的作用。
3.根据权利要求2所述的应用，其特征在于，所述丹参素作为雌激素受体拮抗剂竞争性地与ERα结合，抑制ERα的表达。
4.根据权利要求1所述的应用，其特征在于，所述药物包括口服片剂；所述口服片剂还包括医药领域可接受的辅料。
5.根据权利要求4所述的应用，其特征在于，所述丹参素的有效剂量为90.9～540.9mg/kg。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811286053.9A CN109364055B (zh) | 2018-10-31 | 2018-10-31 | 丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811286053.9A CN109364055B (zh) | 2018-10-31 | 2018-10-31 | 丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109364055A CN109364055A (zh) | 2019-02-22 |
| CN109364055B true CN109364055B (zh) | 2020-10-02 |
Family
ID=65390844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811286053.9A Active CN109364055B (zh) | 2018-10-31 | 2018-10-31 | 丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109364055B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110772634A (zh) * | 2019-12-09 | 2020-02-11 | 上海交通大学医学院附属仁济医院 | 肾胺酶相关活性多肽rp220在制备治疗肾脏疾病的药物中的用途 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1701787A (zh) * | 2004-05-24 | 2005-11-30 | 山东绿叶天然药物研究开发有限公司 | 丹参素在制备治疗脑血管疾病的药物中的应用 |
| CN101112369A (zh) * | 2006-07-28 | 2008-01-30 | 山东绿叶天然药物研究开发有限公司 | 丹参素在制备治疗或预防慢性肾功能衰竭及肾纤维化的药物中的应用 |
| CN105079549A (zh) * | 2015-08-25 | 2015-11-25 | 褚静 | 一种治疗狼疮性肾炎的中药 |
-
2018
- 2018-10-31 CN CN201811286053.9A patent/CN109364055B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1701787A (zh) * | 2004-05-24 | 2005-11-30 | 山东绿叶天然药物研究开发有限公司 | 丹参素在制备治疗脑血管疾病的药物中的应用 |
| CN101112369A (zh) * | 2006-07-28 | 2008-01-30 | 山东绿叶天然药物研究开发有限公司 | 丹参素在制备治疗或预防慢性肾功能衰竭及肾纤维化的药物中的应用 |
| CN105079549A (zh) * | 2015-08-25 | 2015-11-25 | 褚静 | 一种治疗狼疮性肾炎的中药 |
Non-Patent Citations (2)
| Title |
|---|
| 丹参对狼疮性肾炎成纤维细胞增殖、凋亡及c-myc蛋白表达的影响;张国强等;《中国中西医结合杂志》;19970820;第17卷(第8期);473-475 * |
| 丹参注射液有效成分的确定及不同厂家中量效关系的比较;刘倩等;《药物分析杂志》;20140331(第3期);500-504 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109364055A (zh) | 2019-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kuo et al. | STAT3/NF-κB-regulated lentiviral TK/GCV suicide gene therapy for cisplatin-resistant triple-negative breast cancer | |
| DE60129550T2 (de) | Verwendung von niedrigen dosen estrogen in kombination mit immunotherapeutischen verbindungen zur behandlung von immunkrankheiten | |
| KR101926320B1 (ko) | 전립선 암종의 치료를 위한 병용 요법 | |
| US20110085984A1 (en) | Drug Product and Process for Making | |
| WO2021032213A1 (zh) | 靶向组织微环境中衰老细胞的抗衰老药物d/s及其应用 | |
| Rosenblum et al. | Androgenic hormones and human granulopoiesis in vitro | |
| Jia et al. | Digoxin ameliorates joint inflammatory microenvironment by downregulating synovial macrophage M1-like-polarization and its-derived exosomal miR-146b-5p/Usp3&Sox5 axis | |
| CA2534197A1 (en) | Use of a vegf antagonist in combination with radiation therapy | |
| CN109364055B (zh) | 丹参素在制备治疗和/或预防狼疮性肾炎的药物中的应用 | |
| Yang et al. | Preclinical pharmacokinetics, biodistribution, and acute toxicity evaluation of caerin 1.9 peptide in Sprague Dawley rats | |
| TW201517918A (zh) | 癌症放射治療的增敏劑 | |
| Li et al. | Nitazoxanide reduces inflammation and bone erosion in mice with collagen-induced arthritis via inhibiting the JAK2/STAT3 and NF-κB pathways in fibroblast-like synoviocytes | |
| WO2018099363A1 (zh) | 含胰岛素样生长因子-2的药物组合物及其应用 | |
| CN116672338A (zh) | 蟛蜞菊内酯在制备治疗系统性红斑狼疮药物中的应用 | |
| CN107115529B (zh) | Kindlin-2蛋白作为靶点在制备治疗糖尿病的药物中的应用 | |
| Wu et al. | HBSP improves kidney ischemia-reperfusion injury and promotes repair in properdin deficient mice via enhancing phagocytosis of tubular epithelial cells | |
| Cheon et al. | Vigeo attenuates cartilage and bone destruction in a collagen‑induced arthritis mouse model by reducing production of pro‑inflammatory cytokines | |
| CN107569493B (zh) | 氟维司群在制备治疗无功能垂体腺瘤的药物中的用途 | |
| CN115501342B (zh) | 类固醇激素作为孤儿粘附类受体adgrg2拮抗剂配体的应用 | |
| CN114306280B (zh) | 一种雷公藤红素纳米药物及irf1/gstm3轴在制备银屑病药物中的应用 | |
| Zhao et al. | Different cell compositions and a novel somatic KCNJ5 variant found in a patient with bilateral adrenocortical adenomas secreting aldosterone and cortisol | |
| CN118252834B (zh) | 维拉佐酮与瑞戈非尼联合用药在治疗结直肠癌中的应用 | |
| US9301945B2 (en) | Methods for treating inflammatory conditions and states, and cancers by antagonizing NF-κB activation | |
| US20220175795A1 (en) | Use of fulvestrant for the therapeutic care of central core disease | |
| CN118105495A (zh) | 一种增强胆汁酸药物广谱抗病毒效果的方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Application of Danshensu in the preparation of drugs for the treatment and / or prevention of lupus nephritis Effective date of registration: 20211214 Granted publication date: 20201002 Pledgee: Yantai financing guarantee Group Co.,Ltd. Pledgor: BINZHOU MEDICAL University Registration number: Y2021980014859 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Granted publication date: 20201002 Pledgee: Yantai financing guarantee Group Co.,Ltd. Pledgor: BINZHOU MEDICAL University Registration number: Y2021980014859 |