CN109200055A - 一种治疗类风湿关节炎的药物及其用途 - Google Patents
一种治疗类风湿关节炎的药物及其用途 Download PDFInfo
- Publication number
- CN109200055A CN109200055A CN201710553412.1A CN201710553412A CN109200055A CN 109200055 A CN109200055 A CN 109200055A CN 201710553412 A CN201710553412 A CN 201710553412A CN 109200055 A CN109200055 A CN 109200055A
- Authority
- CN
- China
- Prior art keywords
- astragalin
- rheumatoid arthritis
- drug
- injection
- treating rheumatoid
- 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.)
- Pending
Links
- 239000003814 drug Substances 0.000 title claims abstract description 25
- 206010039073 rheumatoid arthritis Diseases 0.000 title claims abstract description 24
- 229940079593 drug Drugs 0.000 title claims abstract description 20
- JPUKWEQWGBDDQB-QSOFNFLRSA-N kaempferol 3-O-beta-D-glucoside Chemical group O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C(C=2C=CC(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O JPUKWEQWGBDDQB-QSOFNFLRSA-N 0.000 claims abstract description 42
- MQVRGDZCYDEQML-UHFFFAOYSA-N Astragalin Natural products C1=CC(OC)=CC=C1C1=C(OC2C(C(O)C(O)C(CO)O2)O)C(=O)C2=C(O)C=C(O)C=C2O1 MQVRGDZCYDEQML-UHFFFAOYSA-N 0.000 claims abstract description 35
- 238000002347 injection Methods 0.000 claims abstract description 7
- 239000007924 injection Substances 0.000 claims abstract description 7
- 239000000546 pharmaceutical excipient Substances 0.000 claims abstract description 3
- 239000000843 powder Substances 0.000 claims abstract description 3
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- 230000002265 prevention Effects 0.000 claims abstract description 3
- -1 pulvis Substances 0.000 claims abstract description 3
- 239000000126 substance Substances 0.000 claims abstract description 3
- 239000003405 delayed action preparation Substances 0.000 claims abstract 2
- 210000002437 synoviocyte Anatomy 0.000 abstract description 14
- 208000024891 symptom Diseases 0.000 abstract description 11
- 108010000684 Matrix Metalloproteinases Proteins 0.000 abstract description 6
- 102000002274 Matrix Metalloproteinases Human genes 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 abstract description 5
- 102000008186 Collagen Human genes 0.000 abstract description 4
- 108010035532 Collagen Proteins 0.000 abstract description 4
- 229920001436 collagen Polymers 0.000 abstract description 4
- 241001465754 Metazoa Species 0.000 abstract description 3
- 241000699670 Mus sp. Species 0.000 abstract description 3
- 230000006698 induction Effects 0.000 abstract 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- 210000000544 articulatio talocruralis Anatomy 0.000 description 13
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 7
- 235000019441 ethanol Nutrition 0.000 description 7
- 229960000485 methotrexate Drugs 0.000 description 7
- 238000002604 ultrasonography Methods 0.000 description 7
- 101710115512 Nuclear receptor coactivator 5 Proteins 0.000 description 6
- 238000004043 dyeing Methods 0.000 description 6
- 230000002757 inflammatory effect Effects 0.000 description 6
- 102000000503 Collagen Type II Human genes 0.000 description 5
- 108010041390 Collagen Type II Proteins 0.000 description 5
- 230000002917 arthritic effect Effects 0.000 description 5
- 229960004756 ethanol Drugs 0.000 description 5
- 230000036039 immunity Effects 0.000 description 5
- 210000000629 knee joint Anatomy 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 210000001179 synovial fluid Anatomy 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 102100027995 Collagenase 3 Human genes 0.000 description 4
- 101000577887 Homo sapiens Collagenase 3 Proteins 0.000 description 4
- 101001013150 Homo sapiens Interstitial collagenase Proteins 0.000 description 4
- 101000990915 Homo sapiens Stromelysin-1 Proteins 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 102000000380 Matrix Metalloproteinase 1 Human genes 0.000 description 4
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 4
- 102100030416 Stromelysin-1 Human genes 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000004945 emulsification Methods 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 3
- 206010003246 arthritis Diseases 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 102000005741 Metalloproteases Human genes 0.000 description 2
- 108010006035 Metalloproteases Proteins 0.000 description 2
- 240000000249 Morus alba Species 0.000 description 2
- 235000008708 Morus alba Nutrition 0.000 description 2
- 101000974353 Mus musculus Nuclear receptor coactivator 5 Proteins 0.000 description 2
- 244000226566 Psoralea corylifolia Species 0.000 description 2
- 241001533115 Thesium chinense Species 0.000 description 2
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 2
- 230000003044 adaptive effect Effects 0.000 description 2
- 210000003423 ankle Anatomy 0.000 description 2
- 229960000190 bacillus calmette–guérin vaccine Drugs 0.000 description 2
- 239000001768 carboxy methyl cellulose Substances 0.000 description 2
- 229940126678 chinese medicines Drugs 0.000 description 2
- 229960000935 dehydrated alcohol Drugs 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 2
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 208000036487 Arthropathies Diseases 0.000 description 1
- 241001061264 Astragalus Species 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108010008165 Etanercept Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000009636 Huang Qi Substances 0.000 description 1
- 208000012659 Joint disease Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 229960002964 adalimumab Drugs 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 235000006533 astragalus Nutrition 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 229960000403 etanercept Drugs 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 229960000598 infliximab Drugs 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 208000018937 joint inflammation Diseases 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000007119 pathological manifestation Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000011268 retreatment Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 210000001258 synovial membrane Anatomy 0.000 description 1
- 210000005222 synovial tissue Anatomy 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 210000004233 talus Anatomy 0.000 description 1
- 210000003684 theca cell Anatomy 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 229940046728 tumor necrosis factor alpha inhibitor Drugs 0.000 description 1
- 239000002452 tumor necrosis factor alpha inhibitor Substances 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及一种治疗类风湿关节炎的药物,其有效成分为紫云英苷,有效浓度为10‑1000μM。紫云英苷与药物学上可接受的药物辅料混合形成散剂、膏剂、粉剂、针剂、水剂、肠溶缓释制剂或注射剂。本发明经动物实验证明,紫云英苷能减轻胶原诱导类风湿关节炎小鼠临床症状评分和关节滑液体积,抑制TNFα诱导人滑膜细胞基质金属蛋白酶的表达,能够有效改善类风湿关节炎症状。本发明还涉及紫云英苷在制备防治类风湿关节炎的药物或保健品中的用途,紫云英苷的有效作用浓度为10‑1000μM。
Description
技术领域
本发明涉及一种治疗类风湿关节炎的药物,并提出该药物在制备各种形式的预防和治疗类风湿关节炎的药物或保健食品中的应用。
背景技术
现有技术公开了类风湿关节炎(Rheumatoid Arthritis,RA)是一种慢性、系统性、进行性、自身免疫性疾病,以广泛的持续存在的关节滑膜炎及对称性、破坏性的关节病变为特征。RA的治疗却非常困难,到目前为止,仍然缺乏安全有效的治疗药物。现代医学主要是抗炎及减轻后遗症,TNFα抑制剂Etanercept、Infliximab和Adalimumab等药物能够将一部分RA患者肿胀、触痛关节计数等客观症状指标减少到50%,但仍然有许多患者在初次治疗时对其不应答,或二次治疗失败,平均每年约有10%的患者因此停药。寻找安全有效的治疗药物成为RA研究亟待解决的重大问题。中医学在治疗RA方面历史悠久,积累了丰富的临床经验,在缓解和改善临床症状方面常能取得较好疗效。
滑膜炎症是RA的主要病理表现,滑膜细胞在炎症因子刺激下,出现滑膜增生,滑液大量产生,产生大量基质金属蛋白酶MMP1、MMP3和MMP13,导致关节软骨的破坏,关节腔增加,因此抑制滑膜细胞基质金属蛋白酶的表达具有重要的治疗意义。
发明内容
针对现有技术的上述不足,根据本发明的实施例,希望提供一种疗效确切的治疗类风湿关节炎的药物。
根据实施例,本发明技术方案提出的一种治疗类风湿关节炎的药物,其有效成分为紫云英苷,有效浓度为10-1000μM。紫云英苷是从百蕊草、桑叶、菟丝子、甘草、补骨脂和蒙古黄芪等中药中提取的有效成分,它可以减轻类风湿关节炎小鼠的症状评分和关节腔间隙,抑制人滑膜细胞基质金属蛋白酶1、3和13的表达。
根据实施例,本发明技术方案的提出是基于紫云英苷对类风湿关节炎具有治疗作用。
根据一个实施例,本发明技术方案的提出是基于紫云英苷作为类风湿关节炎治疗药物,其在患者体液中的有效作用浓度为10-1000μM。
根据一个实施例,本发明技术方案中,紫云英苷可与药物学上可接受的药物辅料混合形成散剂、膏剂、粉剂、针剂、水剂或注射剂。
根据一个实施例,紫云英苷作为类风湿关节炎治疗药物,为主要有效成分制备的各种形式的药物或保健食品,可以用于预防和治疗类风湿关节炎。在使用时可以采取皮下、静脉注射或肛肠给药;注射液的使用可以任意选用生理盐水、葡萄糖、稳定剂、防腐剂、悬浮剂或乳化剂等。
本发明随后的实验例进行了动物实验,建立DBA/1小鼠CIA模型。DBA/1雌鼠6-8周,适应性饲养1周后,采用牛二型胶原(2mg/mL)和完全弗氏佐剂(卡介苗浓度为4mg/ml),完全乳化后,尾部皮下注射100μL(含100μg牛二型胶原),21天后进行二次免疫,采用牛二型胶原(2mg/mL)和不完全弗氏佐剂,完全乳化后,尾部皮下注射100μL(含100μg牛二型胶原)。DBA/1小鼠二次免疫第1天(D20)后,将造模小鼠随机分为组,分为紫云英苷组和模型组,每组10只以上。紫云英苷组中紫云英苷溶解在0.5%的羧甲基纤维素钠(CMC-Na)制备成混悬液,模型组为0.5%的CMC-Na。紫云英苷用量为40mg/kg。灌胃给药,每天一次,连续治疗30天(D50)。取踝关节,进行关节炎症状评分,B超检测踝和膝关节滑液体积,关节切片HE染色观察踝关节炎性浸润情况。经动物实验关节炎症状评分,紫云英苷可以减轻关节炎评分,体内B超检测结果表明,紫云英苷能减轻CIA小鼠踝关节滑液体积和膝关节的滑液体积;HE染色结果显示紫云英苷能减轻CIA小鼠踝关节炎症。本发明还进行了人滑膜细胞体外实验,用TNFα(10ng/mL)加不同浓度紫云英苷(0,10,1000μM)处理人滑膜细胞,24小时后,抽提细胞RNA,用real time PCR方法检测基质金属蛋白酶MMP1,MMP3和MMP13的mRNA表达量,发现TNFα(10ng/mL)显著刺激滑膜细胞表达基质金属蛋白酶,紫云英苷呈剂量依赖性降低上述基质金属蛋白酶的表达量。因此,以紫云英苷为主要有效成分制备的各种形式的药物或保健食品,可以用于预防和治疗类风湿关节炎。
附图说明
图1为紫云英苷的结构简式。
图2动物关节炎症状评分图。
图3是动物实验体内的B超检测结果图。
图4是动物实验体内的HE染色检测结果图。
图5是人滑膜细胞体外实验qPCR检测结果图。
具体实施方式
下面结合附图和具体实施例,进一步阐述本发明。这些实施例应理解为仅用于说明本发明而不用于限制本发明的保护范围。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,这些等效变化和修改同样落入本发明权利要求所限定的范围。
本发明以下实验例中,紫云英苷(结构简式如图1所示)采用市售品或按照中药领域常用提取方法从百蕊草、桑叶、菟丝子、甘草、补骨脂和蒙古黄芪等中药中提取。
本发明以下实施例中,M为摩尔浓度,即mol/L;μM为微摩尔每升。
实验例
建立DBA/1小鼠CIA模型。DBA/1雌鼠6-8周,适应性饲养1周后,采用牛二型胶原(2mg/mL)和完全弗氏佐剂(卡介苗浓度为4mg/ml),完全乳化后,尾部皮下注射100μL(含100μg牛二型胶原),21天后进行二次免疫,采用牛二型胶原(2mg/mL)和不完全弗氏佐剂,完全乳化后,尾部皮下注射100μL(含100μg牛二型胶原)。DBA/1小鼠二次免疫第1天(D20)后,将造模小鼠随机分为组,分为紫云英苷组和模型组,每组10只以上。紫云英苷组中紫云英苷溶解在0.5%的羧甲基纤维素钠(CMC-Na)制备成混悬液,模型组为0.5%的CMC-Na。紫云英苷用量为40mg/kg。灌胃给药,每天一次,连续治疗30天(D50)。取踝关节,进行关节炎症状评分,B超检测踝和膝关节滑液体积,关节切片HE染色观察踝关节炎性浸润情况。本发明还进行了人滑膜细胞体外实验,用TNFα(10ng/mL)加不同浓度紫云英苷(0,10,1000μM)处理人滑膜细胞,24小时后,抽提细胞RNA,用real time PCR方法检测基质金属蛋白酶MMP1,MMP3和MMP13的mRNA表达量。
4.1进行动物实验关节炎症状评分。
关节炎症状评分标准如下,每一周观察一次,统计评分结果。
每只小鼠最低评分0分,最高评分为16分。
如图2所示,实验结果发现,在二次免疫第10天到30天,紫云英苷组的症状评分低于对照组(Astragalin,紫云英苷),效果与甲氨蝶呤类似(MTX,甲氨蝶呤)。
4.2进行B超检测。
动物进行B超检测各组小鼠踝关节和膝关节的滑液体积,B超扫描,并三维重建,图3为网状三维重建图为小鼠踝关节和膝关节滑液的三维重建图。如图3所示,与WT小鼠相比,CIA小鼠踝、膝关节滑液体积显著增加。与CIA组比较,紫云英苷组小鼠踝、膝关节滑液体积明显减轻,效果与西药甲氨蝶呤类似。
4.3进行动物实验的体内HE染色检测。
小鼠踝关节取材后,10%多聚甲醛固定24h,PBS浸泡清洗后,脱钙18天,每2-3天换脱钙液,PBS浸泡清洗后,组织脱水机中18h脱水,石蜡包埋机中浸蜡包埋,矢状位切片(4μm),载玻片平置于烤片机进行贴片5h,垂直置于60℃烤箱8h流蜡后,于通风橱中常规脱蜡至水(二甲苯4minX3→无水乙醇2min→95%乙醇2min→85%乙醇2min→75%乙醇2min→水洗)→苏木素染液1.5min→水洗→2%盐酸乙醇分化5sec→水洗→0.5%氨水返蓝15sec→水洗→伊红染液1.5min→蒸馏水洗→脱水透明(95%乙醇30sec→95%乙醇30sec→95%乙醇3min→无水乙醇30sec→二甲苯4minX3)→中性树胶封片。通风橱中放置4h后显微镜下观察。
如图4所示,在control组小鼠的踝关节,可见伊红染色的骨组织和滑膜组织组成,未见苏木素细胞核染的炎症组织出现。在CIA组小鼠的踝关节,出现大面积苏木素细胞核染的炎症组织浸润。与CIA组比较,紫云英苷组炎症面积明显减少,效果与阳性对照药甲氨蝶呤类似。研究结果提示紫云英苷可以减轻类风湿关节炎的炎症程度。
4.5进行细胞学体外实验,人滑膜细胞炎症因子mRNA表达的检测。
选用人来源的滑膜细胞株,用TNFα(10ng/mL)加不同浓度紫云英苷(0,10,1000μM)处理人滑膜细胞,24小时后,抽提细胞RNA,用real time PCR方法检测基质金属蛋白酶MMP1、MMP3和MMP13的mRNA表达量,发现TNFα(10ng/mL)显著刺激滑膜细胞表达基质金属蛋白酶,紫云英苷呈剂量依赖性降低基质金属蛋白酶的表达量(如图5所示)。
以上结果提示,紫云英苷抑制滑膜细胞基质金属蛋白酶表达,减轻类风湿关节炎小鼠关节炎症,说明紫云英苷是可用于治疗类风湿关节炎的药物。
Claims (4)
1.一种治疗类风湿关节炎的药物,其特征是,有效成分为紫云英苷,有效浓度为10-1000μM。
2.按权利要求1所述的治疗类风湿关节炎的药物,其特征是,紫云英苷与药物学上可接受的药物辅料混合形成散剂、膏剂、粉剂、针剂、水剂、肠溶缓释制剂或注射剂。
3.紫云英苷在制备防治类风湿关节炎的药物或保健品中的用途。
4.根据权利要求3的用途,其特征是,紫云英苷的有效浓度为10-1000μM。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710553412.1A CN109200055A (zh) | 2017-07-08 | 2017-07-08 | 一种治疗类风湿关节炎的药物及其用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710553412.1A CN109200055A (zh) | 2017-07-08 | 2017-07-08 | 一种治疗类风湿关节炎的药物及其用途 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN109200055A true CN109200055A (zh) | 2019-01-15 |
Family
ID=64991550
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710553412.1A Pending CN109200055A (zh) | 2017-07-08 | 2017-07-08 | 一种治疗类风湿关节炎的药物及其用途 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN109200055A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114588152A (zh) * | 2022-04-14 | 2022-06-07 | 安徽医科大学 | Pu.1抑制剂db2313的用途 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1100633A (zh) * | 1993-07-09 | 1995-03-29 | 吴羽化学工业株式会社 | 软骨保护剂 |
CN103110650A (zh) * | 2013-01-29 | 2013-05-22 | 江苏省中国科学院植物研究所 | 紫云英苷在制备抗卵巢衰老药物中的应用 |
-
2017
- 2017-07-08 CN CN201710553412.1A patent/CN109200055A/zh active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1100633A (zh) * | 1993-07-09 | 1995-03-29 | 吴羽化学工业株式会社 | 软骨保护剂 |
CN103110650A (zh) * | 2013-01-29 | 2013-05-22 | 江苏省中国科学院植物研究所 | 紫云英苷在制备抗卵巢衰老药物中的应用 |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114588152A (zh) * | 2022-04-14 | 2022-06-07 | 安徽医科大学 | Pu.1抑制剂db2313的用途 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109200039A (zh) | 治疗类风湿性关节炎的药物及其用途 | |
CN109091477A (zh) | 治疗类风湿性关节炎的小分子化合物及其用途 | |
CN109200047A (zh) | 一种治疗类风湿性关节炎的药物及其用途 | |
CN109200055A (zh) | 一种治疗类风湿关节炎的药物及其用途 | |
CN104382954A (zh) | 一种组合物、用途及保健品 | |
CN106361993A (zh) | 一种防治胃黏膜损伤的药物组合物及其制备方法 | |
CN105497059A (zh) | 一种石斛提取物与铁的复合物及其制备方法与应用 | |
CN102406665A (zh) | 物理改性后的眼镜蛇蛇毒在制备治疗关节炎药物中的应用 | |
CN109096360B (zh) | 一种治疗类风湿性关节炎的小分子化合物及其用途 | |
CN104083528A (zh) | 一种提高人体免疫力的褐藻姬松茸复合片及其制备方法 | |
CN103655849B (zh) | 治疗风寒感冒的中药组合物及其制备方法和应用 | |
CN102657827A (zh) | 治疗肝胆疾病的中药及其制备方法 | |
CN104623382A (zh) | 一种治疗痰热内扰型失眠的中药 | |
CN101095708B (zh) | 柴胡总多糖在制备防治系统性红斑狼疮药物中的用途 | |
CN103961341B (zh) | 一种治疗偏头痛的药物组合物及其应用 | |
CN109078012A (zh) | 黄芩素在制备防治伊立替康诱导的化疗性肠炎药物中的应用 | |
CN103705500B (zh) | Myrtucommuacetalone在治疗肾功能不全药物中的应用 | |
CN109999143B (zh) | 一种治疗良性肿瘤增生性疾病的中药 | |
CN101647841A (zh) | 地胆草及提取物的新应用 | |
CN107213254A (zh) | 复方血栓通制剂在制备治疗paf介导疾病药物中的用途 | |
CN104042696B (zh) | 一种治疗胆汁反流性胃炎的中药提取物及其用途 | |
CN105213967A (zh) | 一种治疗恶性肿瘤的中药制剂 | |
CN104622987A (zh) | 一种治疗慢性肝炎肝硬化的药物组合物及应用 | |
CN105194453A (zh) | 一种治疗小儿积食的中药制剂 | |
CN103356689B (zh) | Polyflavanostilbene A在制备治疗肾功能不全药物中的应用 |
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 | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190115 |
|
RJ01 | Rejection of invention patent application after publication |