CN101180044B - 用于自身免疫损伤后中枢神经系统神经再生的egf/ghrp-6组合 - Google Patents
用于自身免疫损伤后中枢神经系统神经再生的egf/ghrp-6组合 Download PDFInfo
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Abstract
本发明涉及在自身免疫损伤后刺激中枢神经系统的神经再生。更具体而言,本发明涉及包括治疗有效浓度的表皮生长因子和人生长激素促分泌六肽的药物组合,将其给药于罹患多发性硬化和视神经脊髓炎症状的患者,其可纠正中枢神经系统中自体反应性细胞引起的脱髓鞘。
Description
发明领域
本发明涉及药物,更具体地,涉及神经学,并涉及自身免疫损伤后刺激中枢神经系统神经再生,特别是通过给药含有表皮生长因子和生长激素释放肽-6的组合物,用于治疗和预防患有多发性硬化和视神经脊髓炎患者的复发。
背景技术
多发性硬化(MS)和视神经脊髓炎(NO)是影响年轻人主要是女性的自身免疫脱髓鞘疾病,产生的机能不全和衰弱随着时间向不利方向演化。MS发病率与第一世界国家的产业化和发展的先进参数强烈相关。中枢神经系统是一个有特权的免疫学部位,其中很少发现自身免疫反应。这在由于不确定的原因细胞和体液调节机制失败时发生,其决定了随着激活的淋巴细胞穿过血脑屏障(BBB)并发现其在中枢神经系统实质内的靶标,外周产生的自体反应性细胞对抗髓磷脂抗原(这是时常发生的情况)。多种级联样事件跟着发生,发展成脱髓鞘、反应性星形细胞增生以及神经元和少突胶质细胞死亡。
中枢神经系统内的自身免疫反应是针对髓磷脂抗原,使得最初损伤限于包裹主要神经元轴突的髓磷脂层、负责髓磷脂产生的少突胶质细胞,由于自身免疫反应的扩展其它组的神经元受到非特异性损伤。
随后发生的脱髓鞘和坏死或凋亡引起的神经元死亡导致随机影响人体内多种结构的运动和感觉丧失。MS和ON中的髓鞘再生过程通常是有限而短暂的。尽管这种髓鞘再生过程是可能的,其取决于自体反应性星形细胞和少突胶质细胞之间的平衡(John G.R.,Shankar S.L.,Shafit-Zagardo B.,Massimi A.,Lee S.C,Raine C.S.等(2002)Multiple sclerosis:re-expression of a developmental pathwaythat restricts oligodendrocyte maturation Nature Medicine8(10):1115-1121)。
神经再生策略中,用生长因子如表皮生长因子(EGF)和牛成纤维细胞生长因子(bFGF)进行治疗已经成为非常有前景的提议,证明了从大脑皮层分离的多能未分化细胞世系对这些生长因子有反应,向不同的细胞世系如I型和II型星形细胞、有髓鞘的少突胶质细胞和不同的神经元世系分化(Mehler M.F.,Gokhan S.(1999)Postnatalcerebral cortical multipotent progenitors:regulatorymechanisms and potential role in the development of novel neuralregenerative strategies.Brain Pathol;9(3):515-526)。
生长激素释放肽-6(GHRP-6)增加中枢神经系统中胰岛素样生长因子1(IGF-1)表达(Frago L.M.,Paneda C.,Dickson S.L.,HewsonA.K.,Argente J.,Chowen J.A.(2002)Growth hormone(GH)andGH-releasing peptide-6 increase brain insulin-like growthfactor-I expression and activate intracellular signalingpathways involved in neuroprotection Endocrinology143(10):4113-4122)。IGF-1涉及以下过程,如少突胶质细胞成熟(Wilson H.C.,Onischke C.,RaineC.S.(2003)Humanoligodendrocyte precursor cells in vitro:phenotypic analysisand differential response to growth factors.Glia44(2):153-165)、在MS和ON中中止依赖于TNF-α激活的凋亡通路同时保护免受该因子诱导的损伤(Ye P,D′Ercole A.J.(1999)Insulin-like growth factor 1 protects oligodendrocytes fromtumor necrosis factor-alpha-induced injury Endocrinology140(7):3063-3072)。IGF-1还下调I类MHC相关分子的表达(Ito T,Ito N,Bettermann A,Tokura Y,Takigawa M,Paus R.(2004)Collapse and restoration of MHC class-I-dependent immuneprivilege:exploiting the human hair follicle as a model Am.J.Pathol164(2):623-634)。在EAE的动物模型中,IGF-1减轻BBB的血管内皮损伤,减少硬化斑(MS的标志损伤)的数目和大小(Li W,Quigley L,Yao D.L,Hudson L.D,Brenner M,Zhang B.J et al.(1998)Chronic relapsing experimental autoimmune encephalonyelitis:effects of insulin-like growth factor-1 treatment on clinicaldeficits,lesion severity,glial responses,and blood brainbarrier defects.J Neuropathol Exp Neurol 57(5):426-438)。
全身给药时,GHRP-6提高内源促肾上腺皮质激素(ACTH)的水平(Martins M.R,Pinto A.C,Brunner E,Silva M.R,Lengyel A.M.(2003)GH-releasing peptide(GHRP-6)-induced ACTH release inpatients with addison′s disease:effect of glucocorticoidwithdrawal.JEndocrinol Invest.26(2):143-147)。ACTH作为内源类固醇释放因子对抵抗自体反应性疾患具有有益的效果并且已经长时间成为对MS的传统治疗(Oishi C,Sakuta M.(2003)Steroidtherapy for multiple sclerosis.Nippon Rinsho61(8):1361-1366)。
EGF在中枢神经系统(CNS)中由小胶质细胞、血液来源的巨噬细胞以及一些神经元局部合成。因为其通过BBB和脑室底膜,EGF能够流过CNS。已将若干特定的生理功能归于EGF,如CNS发育、CNS实质细胞的维持和分化,这些作用与神经再生过程和损伤时引发的生存机制非常相关(Plata-Salaman C.R.(1991)Epidermal growth factorand the nervous system Peptides 12(3):653-663)。
EGF刺激CNS内的细胞增殖和存活(Thorne R.G,Hrabetova S,Nicholson C.(2004)Diffusion of Epidermal Growth Factorin RatBrain Extracellular Space Measured by Integrative OpticalImaging J Neurophysiol 92(6):3471-3481)。
经EGF刺激的少突胶质细胞获得增强的髓鞘再生潜能。EGF有助于少突胶质细胞增殖过程,因此促进细胞分裂的开始和进一步分化成特化细胞如成熟少突胶质细胞、星形细胞和施万细胞。EGF促进诸如新神经元产生引起的神经形成这样的事件(Crang A.J.,Gilson J.M.,Li W.W.,Blakemore W.F.(2004)The remyelinating potential andin vitro differentiation of MOG-expressing oligodendrocyteprecursors isolated from the adult rat CNS.Eur J Neurosci20(6):1445-1460;Raineteau 0.,Rietschin L.,Gradwohl G.,Guillemot F.,Gahwiler B.H.(2004)Neurogenesis in hippocampalslice cultures.Mol Cell Neurosci 26(2):241-250)。在少突胶质细胞经历损伤后可能看到更多的这些事件,表明EGF-介导的对再生过程的作用是EGF与损伤的少突胶质细胞中被特异性活化的信号转导系统之间相互作用的结果。这还表明对这种信号转导系统的调节可以放大朝向髓鞘再生的机制(Wang K.,Wang J..J,Wang Y.,He Q.H.,Wang X.,Wang X.M.(2004)Infusion of epidermal growth factorand basic fibroblast growth factor into the striatum ofparkinsonian rats leads to in vitro proliferation anddifferentiation of adult neural progenitor cells.Neurosci Lett364(3):154-158;Knapp P.E.,Adams M.H.(2004)Epidermal growthfactor promotes oligodendrocyte process formation and regrowthafter injury.Exp Cell Res 296(2):135-144)。
在最近几年中,已经提出复杂的治疗干预作为联合治疗和/或系统治疗,其远不显得多余并强化治疗途径,因此使得可以在与所提及的疾病相关的节点或关键点接近复杂的病理生理学问题。仍然缺乏与多种生长因子的联合治疗或它们中的一种与备选的对CNS具有正面营养作用的分子的组合。
理想的治疗将集中于遏制与最初的发作相关的症状强度并将复发频率减至最小。因此,其支持对开发用于不同临床形式MS和NO的治疗和复发预防的更有效方法的兴趣。
之前已经提出治疗有效浓度的EGF和促分泌素GHRP-6的组合的给药用于预防和治疗由于动脉血供不足而致的组织损伤(WO02/053167)。
发明概述
本发明是基于一种方法,其中EGF和GHRP-6的共同给药代表改进的对CNS的自身免疫疾病的治疗。这种组合保护并恢复CNS慢性过程中自身免疫相关的损伤,特别是在多发性硬化和视神经脊髓炎中。
与单独的每种成分相比较,该组合产生更持久的功效和复发的实质性降低-换句话说,它以更有效的方式触发了再生事件。如在此所用的,术语“更持久的功效”意思是活性成分导致MS和ON相关症状更长时间的缓解,甚至赋予保护来避免复发发作。这将确保作为自身免疫损伤引起的脱髓鞘和凋亡/坏死所致的神经元丧失的结果的受影响神经功能的恢复。另一方面,药物组合中的活性成分是具有天然调节细胞刺激能力的自体蛋白质和肽,因此它们在外源肽施用后能受到刺激而增殖。通过这种方式,可以抵抗自身免疫应答,因为调节T细胞的作用是组成性地介导免疫耐受(Jrn G.,Benedikt B.,Bruno K.(2004)Medullary Epithelial Cells of the Human Thymus Expressa Highly Diverse Selection of Tissue-specific Genes Colocalizedin Chromosoma l Clusters.J Exp Med 199(2):155-166.);(DayneM.,Christophe B.(2004)Back to Central Tolerance.Immunity20:509-516);(Mark S.A.,Emily S.V.,Ludger K.,Zhibin C.,StuartP.B.,Shannon J.T.et al.(2002)Projection of anImmunological Self Shadow Within the Thymus by the Aire Protein.Science 298:1395-1400)(Shimon S.(2004)Naturallyarising CD4+regulatory T cells for immunologic self-tolerance and negativecontrol of immune responses.Annual Review of Immunology22:531-562)。
由于EGF和GHRP-6之间在神经营养和神经再生事件方面的协同效应,这种组合可用于加速神经形成过程,这有助于重新获得由于自身免疫产生的损伤而丧失的神经功能。EGF和GHRP-6的组合可以结合任何抗氧化治疗。针对神经再生和神经保护的所述组合的治疗性给药需要重复的给药方案。如本发明中所述的,称为EGF的活性成分可以源自任何动物物种,包括羊、牛、猪和人,以其天然序列或其变体,并可来自任何来源,如合成、天然或重组的。这种情况中的优选形式是天然序列的人EGF,尤其是人重组EGF。称为生长激素释放肽(GHRP)的活性成分是具有以下序列的六肽:His-D-Trp-Ala-Trp-D-Phe-Lys-NH2,通过化学合成获得。
在特定的情况中,在MS和ON危象期间给药的治疗剂量对于EGF为5-200μg/kg/天,对于GHRP-6为0.5-350μg/kg/天,持续20-30天。
在本发明的另一种情况中,在多发性硬化中为了预防复发,危象间期内给药的剂量对于每种成分为0.5-50μg/kg/天,持续至多130天。该组合必须以推注(bolus)来给药。给药途径为非肠道-外周静脉内、肌内或腹膜内。给药中涉及的载体包括生理盐水溶液、乳酸林格溶液、人血浆、人白蛋白溶液、5%葡萄糖、明胶溶液或其混合物。
期待获得对复发缓解型或继发进展型临床形式的多发性硬化的最高疗效,第一次给药必须与疾病的前驱症状同时进行(个人化的)。在缓解期,推荐用上述剂量持续治疗方案作为对复发的预防。
在其他多发性硬化临床形式中,推荐包括治疗剂量的持续治疗方案。
根据本发明的其他情况,组合EGF/GHRP-6在过继转移实验中诱导防止EAE严重临床形式发作的天然和适应性调节T细胞的增殖。
组合EGF/GHRP-6可以于单一药物组合物中使用或就在使用前混合单独的成分。可以通过缓释装置来使用活性成分组合。如果制剂是冻融的,必须就在使用前稀释。
特定实施方案/实施例的详述
实施例1.EGF/GHRP-6药物组合在实验性自身免疫脑炎(EAE)生物模型中的治疗效果
为了评价EGF/GHRP-6药物组合的治疗功效,建立了EAE动物模型,其代表多发性硬化人类疾病的动物形式。
在第0和6天,用在PBS(50%)和完全弗氏佐剂(50%)中的豚鼠脊髓匀浆(5mg)皮下免疫雌性Lewis大鼠(130g)。第一次免疫后十天,使用组合EGF/GHRP-6(200μg/kg/天-740μg/kg/天)、单独的活性成分EGF(200μg/kg/天)、GHRP-6(740μg/kg/天)和安慰剂(PBS)来启动治疗方案。遵循该治疗方案10天,使用腹膜内给药。临床分数是基于以下的分级:0;无症状,1;尾部瘫痪,2;任何后肢瘫痪,3;后肢完全瘫痪,4;前肢和后肢完全瘫疾,5;垂死或死亡。体重减轻以及膀胱和直肠括约肌失禁(其也是疾病的临床征象),通过向之前所述的临床指数加0.5来评分。第一次免疫后四十天,将动物麻醉并实施安乐死,将脑和脊髓进行加工处理用于组织病理学研究(10%福尔马林,H&E和Luxol Blue染色)。对于组织病理学分析,考虑以下参数:血管周围炎性浸润的数量和大小、脱髓鞘损伤的数量、凋亡神经元的数量以及神经胶质细胞和星形细胞反应性。以盲法进行显微镜研究。
如表1中所示的,组合EGF/GHRP-6保护经实验诱导发生EAE的动物,只有50%的这些动物发生最轻形式的疾病,而剩余的保持未改变。相比之下,在剩余的组中,疾病具有100%的发病率(用单独的活性成分和安慰剂治疗的组)。EGF/GHRP-6组合治疗动物中的平均临床指数为0.37±0.47。对于单独成分治疗组,平均临床指数对于EGF为1.37±1.7,对于GHRP-6为1.5±1.6,安慰剂治疗组显示出平均临床指数1.7±1.4。每个组分配八只大鼠。组间的统计学比较为p<0.001。使用了Newman Keuls多重比较检验。
表1.对诱导发生EAE的大鼠的治疗效果的临床概述
组 | 发病率(%) | 初次出现的天数(平均值±SD) | 临床分数 | |||
平均值±SD | 最大值 | 最小值 | 疾病持续时间 | |||
对照 | 0 | 0 | 0 | 0 | 0 | 0 |
EGF | 100 | 12.5±0.57 | 1.37±1.7 | 5 | 1 | 12.7±4.1 |
GHRP-6 | 100 | 13.7±2.3 | 1.5±1.6 | 4 | 0.5 | 15.5±6.7 |
EGF/GHRP-6 | 50 | 12±0 | 0.37±0.47 | 1 | 0 | 9±1.4 |
安慰剂 | 100 | 12.2±0.5 | 1.7±1.4 | 4 | 0.5 | 13.2±8.1 |
如表2中所示的,对不同组中包括的动物的脑和脊髓的病理学研究分析的结果表明,即使对于反应性星形细胞存在相同的情况,EGF/GHRP-6治疗的动物中血管套(cuff)的数量(p=0.028不成对T检验)和大小小于安慰剂治疗组。
表2.动物脑和脊髓中的血管周围炎性浸润
组 | 血管周围炎性浸润的数量(平均值±SD) |
对照 | 0 |
EGF | 4.5±1.9 |
GHRP-6 | 2.25±1.25 |
EGF/GHRP-6 | 2±0.8 |
安慰剂 | 5±2.1 |
该实验证明了EGF/GHRP药物组合保护动物免于发生严重临床形式的疾病。该保护作用所潜在的机制是少突胶质细胞产生髓磷脂增加和随后受累神经结构的髓鞘再生。另一种机制与BBB的完整性相关,避免自体反应性细胞通过进入脑实质。
实施例2.EAE动物模型预防方案中EGF/GHRP-6药物组合的保护效果
为了评估EGF/GHRP-6组合在代表MS人类疾病的EAE动物模型中的预防作用,在第0和6天,用在PBS(50%)和完全弗氏佐剂(50%)中的豚鼠脊髓匀浆(5mg)皮下免疫雌性Lewis大鼠(130g)。在第一次免疫前十天,使用EGF/GHRP-6组合(200μg/kg/天-740μg/kg/天)和分别单独的活性成分EGF(200μg/kg/天)、GHRP-6(740μg/kg/天)和安慰剂(PBS)启动预防方案。遵循该预防方案10天(在第一次免疫之前-10至-1天),使用腹膜内给药途径。临床分数是基于以下的分级:0;无症状,1;尾部瘫痪,2;任何后肢瘫痪,3;后肢完全瘫痪,4;前肢和后肢完全瘫痪,5;垂死或死亡。体重减轻以及膀胱和直肠括约肌失禁(其也是疾病的临床征象),通过向之前所述的临床指数加0.5来评分。第一次免疫后四十天,将动物麻醉并实施安乐死,将脑和脊髓进行加工处理用于组织病理学研究(10%福尔马林,H&E和Luxol Blue染色)。对于组织病理学分析,考虑以下参数:血管周围炎性浸润的数量和大小、脱髓鞘损伤的数量、凋亡神经元的数量以及神经胶质细胞和星形细胞反应性。以盲法进行显微镜研究。
如表3中所示的,预防方案中使用的EGF/GHRP-6药物组合保护经诱导发生EAE的动物。100%EGF/GHRP-6治疗的动物发生轻微形式的疾病(0.5-1临床分数)。相比之下,对于用单一成分治疗的组,75%发生严重临床形式的EAE(3-4临床分数)。发现用药物组合EGF/GHRP-6治疗的动物的平均临床指数为0.68±0.25。对于单一成分治疗的组,对于EGF平均临床指数为2.8±0.99,对于GHRP-6为2.7±1.03(与EGF/GHRP-6治疗的动物相比较,P=0.0003),安慰剂治疗组显示出3±1.4的平均临床指数(与EGF/GHRP-6治疗的动物相比较,p=0.0011)。每组使用八只大鼠。对于统计学比较,使用Mann Whitney T检验,比较以组合治疗的组和安慰剂组,获得p=0.0011的值;与用药物组合治疗的组相比较,对用单独的活性成分治疗的组获得p=0.0003的值。
表3.在诱导发生EAE的大鼠中的预防效果的临床概述
组 | 发病率(%) | 初次出现的天数(平均值±SD) | 临床分数 | ||
平均值±SD | 最大值 | 最小值 | |||
对照 | 0 | 0 | 0 | 0 | 0 |
EGF | 100 | 11.5±0.57 | 2.8±0.99 | 4 | 1 |
GHRP-6 | 100 | 11.7±2.3 | 2.7±1.03 | 4 | 1 |
EGF/GHRP-6 | 100 | 10±0 | 0.68±0.75 | 1 | 0.5 |
安慰剂 | 100 | 12.2±0.5 | 3±1.4 | 5 | 1 |
如表4中所观察到的,对实验组中脑和脊髓的病理学分析表明,即使在反应性星形细胞相同的情况中,EGF/GHRP-6预防性治疗的动物中血管周围套的数量(p=0.025不成对T检验)和大小比安慰剂治疗组要少而小。
表4.预防性治疗的动物的脑和脊髓中的血管周围炎性浸润
组 | 血管周围炎性浸润的数量(平均值±SD) |
对照 | 0 |
EGF | 4±1.6 |
GHRP-6 | 2.7±1.95 |
EGF/GHRP-6 | 1.5±1 |
安慰剂 | 5±2.1 |
该实验证明了预防性使用的EGF/GHRP-6药物组合保护动物免于发生最严重临床形式的EAE。此外,证明了临床演变与组织学发现之间强烈的关联。解释这种保护效果的机理与诱导神经元前体细胞朝向少突胶质细胞分化相关,少突胶质细胞在髓磷脂产生中将受到预调节并有活性。BBB完整性的保存将防止自体反应性细胞通过进入脑实质,这是解释预防性使用的EGF/GHRP-6药物组合的保护性作用的另一事件。
实施例3.剂量研究,药物组合的活性成分之间的协同-增强作用
寻找药物组合物对于治疗作用有效的剂量范围,将所述组合用于上述EAE模型中。在第0和6天,用在PBS(50%)和完全弗氏佐剂(50%)中的豚鼠脊髓匀浆(5mg)皮下免疫雌性Lewis大鼠(130g)。第一次免疫后十天使用不同浓度的EGF/GHRP-6组合启动治疗方案。
EGF/GHRP-6(400μg/kg/天-1480μg/kg/天).
EGF/GHRP-6(200μg/kg/天-740μg/kg/天).
EGF/GHRP-6(100μg/kg/天-340μg/kg/天)
EGF/GHRP-6(50μg/kg/天-170μg/kg/天)
EGF/GHRP-6(25μg/kg/天-85μg/kg/天)
EGF/GHRP-6(12μg/kg/天-40μg/kg/天)
遵循该治疗方案10天,使用腹膜内给药。临床分数是基于以下的分级:0;无症状,1;尾部瘫痪,2;任何后肢瘫痪,3;后肢完全瘫痪,4;前肢和后肢完全瘫痪,5;垂死或死亡。体重减轻以及膀胱和直肠括约肌失禁(其也是疾病的临床征象),通过向之前所述的临床指数加0.5来评分。第一次免疫后四十天,将动物麻醉并实施安乐死,将脑和脊髓进行加工处理用于组织病理学研究(10%福尔马林,H&E和Luxol Blue染色)。对于组织病理学分析,考虑以下参数:血管周围炎性浸润的数量和大小、脱髓鞘损伤的数量、凋亡神经元的数量以及神经胶质细胞和星形细胞反应性。以盲法进行显微镜研究。
在EGF/GHRP-6(400μg/kg/天-1480μg/kg/天)治疗组中,25%的动物保持未改变,75%的动物显示出轻微形式的疾病(0.5-1)。平均临床指数是0.62±0.44。在EGF/GHRP-6(200μg/kg/天-740μg/kg/天)治疗组中,25%的动物保持未改变,75%的动物显示出轻微形式的疾病(0.5-1)。平均临床指数是0.5±0.37。
在EGF/GHRP-6(100μg/kg/天-340μg/kg/天)治疗组中,12.5%的动物保持未改变,87.5%的动物显示出轻微形式的疾病(0.5-1)。平均临床指数为0.62±0.35。
在EGF/GHRP-6(50μg/kg/天-170μg/kg/天)治疗组中,100%的动物发生了EAE,12.5%具有中等临床形式(2),剩余的显示出轻微形式的疾病(0.5-1)。平均临床指数是0.93±0.49。
在EGF/GHRP-6(25μg/kg/天-85μg/kg/天)治疗组中,100%的动物发生了EAE,37.5%具有中等临床形式(2),剩余的显示出轻微形式的疾病(0.5-1)。平均临床指数是1.25±0.65。
在EGF/GHRP-6(12μg/kg/天-40μg/kg/天)治疗组中,100%的动物发生了EAE,12.5%具有最严重的临床形式(3),37.5%具有中等临床形式(2),剩余的显示出轻微形式的疾病(0.5-1)。平均临床指数是1.37±0.87。
表5和6显示了临床和组织病理学结果。组织病理学分析表明,在诱导EAE并用EGF/GHRP-6(400μg/kg-1480μg/kg,200μg/kg-740μg/kg,100μg/kg-340μg/kg,50μg/kg-170μg/kg和25μg/kg-85μg/kg)治疗的组中淋巴细胞血管周围浸润的数量无统计学差异。在用EGF/GHRP-6药物组合(12μg/kg-40μg/kg)治疗的组的情况中,血管周围浸润的数量显示出更高的趋势(p=0.040),但是该差异没有统计学显著性。这些结果证明了对于EGF存在50-400μg/kg/天的剂量范围和对于GHRP-6存在170μg/kg/天-1.4mg/kg/天的剂量范围,其使得能配制维持其对抗EAE诱导的保护作用的效用的组合(表5)。
表5.剂量应答和EGF/GHRP-6组合的单独成分之间的协同-增强作用的临床概述
组EGF/GHRP-6 | 发病率(%) | 临床分数 | ||
平均值±SD | 最大值 | 最小值 | ||
EGF/GHRP-6(400μg/kg-1480μg/kg) | 75 | 0.62±0.44 | 0 | 1 |
EGF/GHRP-6(200μg/kg-740μg/kg) | 75 | 0.5±0.37 | 0 | 1 |
EGF/GHRP-6(100μg/kg-340μg/kg) | 87.5 | 0.65±0.35 | 0 | 1 |
EGF/GHRP-6(50μg/kg-170μg/kg) | 100 | 0.93±0.49 | 0.5 | 2 |
EGF/GHRP-6(25μg/kg-85μg/kg) | 100 | 1.25±0.65 | 0.5 | 2 |
EGF/GHRP-6(12μg/kg-40μg/kg) | 100 | 1.3±0.87 | 0.5 | 3 |
表6.每个实验组的脑和脊髓中的血管周围炎性浸润
组EGF/GHRP-6 | |
血管周围炎性浸润的数量(平均值±SD) | |
EGF/GHRP-6(400μg/kg-1480μg/kg) | 2.7±0.95 |
EGF/GHRP-6(200μg/kg-740μg/kg) | 2.7±0.5 |
EGF/GHRP-6(100μg/kg-340μg/kg) | 2.2±1.2 |
EGF/GHRP-6(50μg/kg-170μg/kg) | 3±1.4 |
EGF/GHRP-6(25μg/kg-85μg/kg) | 3.2±0.95 |
EGF/GHRP-6(12μg/kg-40μg/kg) | 4±0.8 |
实施例4.对EGF-GHRP-6药物组合在产生天然调节T细胞应答中的作用的评价
在第0和6天,用在PBS(50%)和完全弗氏佐剂(50%)中的豚鼠脊髓匀浆(5mg)皮下免疫二十只雌性Lewis大鼠(130g)。第一次免疫后十天,在十只经免疫大鼠中通过腹膜内给药使用EGF/GHRP-6(200μg/kg/天-740μg/kg/天)组合开始治疗方案并再进行10天(组A)。另外10只大鼠用作为安慰剂的PBS处理(组B)。最后一次给药后一周,将两个组的动物麻醉用于放血,由此获得外周血单核淋巴细胞。处理源自组A和B的淋巴细胞用于分离CD4+细胞。
通过FACS分析CD4+CD25+细胞,组A中为14.67%,PBS处理组(组B)中为3.8%。
在第0和6天,用在PBS(50%)和完全弗氏佐剂(50%)中的豚鼠脊髓匀浆(5mg)皮下免疫其他雌性Lewis大鼠(130g)。第一次免疫后十天,对一个亚组(n=8)从组A静脉内过继转移500000个CD4+细胞。另一个亚组(n=8)从组B静脉内过继转移500000个CD4+细胞。
临床分数是基于以下的分级:0;无症状,1;尾部瘫痪,2;任何后肢瘫痪,3;后肢完全瘫痪,4;前肢和后肢完全瘫痪,5;垂死或死亡。体重减轻以及膀胱和直肠括约肌失禁(其也是疾病的临床征象),通过向之前所述的临床指数加0.5来评分。第一次免疫后四十天,将动物麻醉并实施安乐死,将脑和脊髓进行加工处理用于组织病理学研究(10%福尔马林,H&E和Luxol Blue染色)。对于组织病理学分析,考虑以下参数:血管周围炎性浸润的数量和大小、脱髓鞘损伤的数量、凋亡神经元的数量以及神经胶质细胞和星形细胞反应性。以盲法进行显微镜研究。
如表7中所示的,在诱导EAE并用药物组合治疗的动物中CD4+细胞的转移保护宿主免于发生EAE。只有50%转移了源自组A的CD4+细胞的动物发生了轻微临床形式的EAE(0.5-1临床分数),剩余的50%保持未改变。相比之下,100%过继转移了源自组B的CD4+细胞的动物发生了EAE,62.5%具有严重临床形式(2-4临床分数),37.5%具有轻微形式(0.5-1临床分数)。在过继转移了源自组A的CD4+细胞的亚组中,平均临床指数是0.31±0.34。在过继转移了源自组B的CD4+细胞的亚组中,平均临床指数是2.1±0.99(P=0.0003)。使用Mann WhitneyT检验进行统计学分析。
表7.过继细胞转移的保护作用
组 | 发病率(%) | 初次出现的天数(平均值±SD) | 临床分数 | ||
平均值±SD | 最大值 | 最小值 | |||
组A | 50 | 10±0 | 0.31±0.34 | 1 | 0 |
组B | 100 | 11.5±0.57 | 2.1±0.99 | 4 | 1 |
如表8中所示的,对实验组中脑和脊髓的组织学分析表明,即使关于反应性星形细胞存在相同的情况,对于源自组A的CD4+细胞,宿主大鼠中血管套的数量(p=0.0001)和大小小于过继转移了源自组B的CD4+细胞的大鼠。
表8.每个实验组的脑和脊髓中的血管周围炎性浸润
组 | 血管周围炎性浸润的数量(平均值±SD) |
组ACD4<sup>+</sup>宿主 | 2.1±0.8 |
组BCD4<sup>+</sup>宿主 | 6.5±1.7 |
这些结果证明了用所述药物组合治疗能够诱导天然调节T细胞的增殖,其在过继转移实验中保护免于发生严重临床形式的EAE。
Claims (8)
1.EGF和生长激素释放肽-6(GHRP-6)用于制备治疗和缓解罹患与脱髓鞘、神经元变性和自身免疫病因的调亡或坏死所致的神经元细胞死亡相关的症状或并发症的患者的中枢神经系统疾病的药物的用途。
2.根据权利要求1的用途,其中EGF是人EGF。
3.根据权利要求2的用途,其中人EGF从天然来源、通过重组技术或化学合成获得。
4.根据权利要求1-3任一项的用途,其中中枢神经系统疾病是:
a.多发性硬化;
b.视神经脊髓炎。
5.根据权利要求1-3任一项的用途,其中EGF和GHRP-6的组合通过静脉内、肌内或腹膜内给药或使用控释装置给药。
6.根据权利要求1-3任一项的用途,其中在多发性硬化和视神经脊髓炎危象期间给药所述药物,剂量范围为对于EGF 5-200μg/kg/天,对于GHRP-60.5-350μg/kg/天,持续20-30天。
7.根据权利要求1-3任一项的用途,其中在多发性硬化危象间期内给药所述药物,剂量范围为对于每种成分0.5-50μg/kg/天,持续至多130天,以预防复发。
8.根据权利要求1的用途,其中所述药物诱导天然和适应性调节T细胞的增殖。
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---|---|---|---|---|
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Non-Patent Citations (10)
Title |
---|
Carlos R. Plata-Salaman.Epidermal growth factor and the nervous system.peptides12 3.1991,12(3),653-663. |
Carlos R. Plata-Salaman.Epidermal growth factor and the nervous system.peptides12 3.1991,12(3),653-663. * |
Knapp P.E et al..Epidermal growth factor promotes oligodendrocyte processformation and regrowth after injury.Experimental cell research296 2.2004,296(2),135-144. |
Knapp P.E et al..Epidermal growth factor promotes oligodendrocyte processformation and regrowth after injury.Experimental cell research296 2.2004,296(2),135-144. * |
kuhn H G et al..epidermal growth factor and fibroblast growth factor-2 havedifferent effects on neural progenitors in the adult rat brain.journal of neuroscience17 15.1997,17(15),5822-5824. |
kuhn H G et al..epidermal growth factor and fibroblast growth factor-2 havedifferent effects on neural progenitors in the adult rat brain.journal of neuroscience17 15.1997,17(15),5822-5824. * |
Laura M.F et al..Growth hormone(GH) and GH-Releasing peptide-6 increasebrain insulin-like growth factor-I expression and activateintracellular signaline pathways involved in neuroprotection.Endocrinology143 10.2002,143(10),4113-4122. |
Laura M.F et al..Growth hormone(GH) and GH-Releasing peptide-6 increasebrain insulin-like growth factor-I expression and activateintracellular signaline pathways involved in neuroprotection.Endocrinology143 10.2002,143(10),4113-4122. * |
Olivier Raineteau et al..Neurogenesis in hippocampal slice cultures.Mol. Cell. Neurosci.26 2.2004,26(2),246-248. |
Olivier Raineteau et al..Neurogenesis in hippocampal slice cultures.Mol. Cell. Neurosci.26 2.2004,26(2),246-248. * |
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EP1870106A2 (en) | 2007-12-26 |
ES2327562T3 (es) | 2009-10-30 |
DE602006006903D1 (de) | 2009-07-02 |
KR101255200B1 (ko) | 2013-04-23 |
KR20070107790A (ko) | 2007-11-07 |
BRPI0607844B8 (pt) | 2021-05-25 |
CU23529A1 (es) | 2010-06-17 |
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