CN1523997B - 稳定的白介素2 - Google Patents
稳定的白介素2 Download PDFInfo
- Publication number
- CN1523997B CN1523997B CN018144454A CN01814445A CN1523997B CN 1523997 B CN1523997 B CN 1523997B CN 018144454 A CN018144454 A CN 018144454A CN 01814445 A CN01814445 A CN 01814445A CN 1523997 B CN1523997 B CN 1523997B
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- Prior art keywords
- histidine
- preparation
- stable
- sucrose
- glycine
- Prior art date
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Abstract
一种稳定的药物制剂，它含有人白细胞介素-2或其变体和稳定有效量的组氨酸。优选的的制剂包含甘氨酸和蔗糖并且IL-2变体具有一个突变，N88R。优选的制剂是冻干形式，它在用水稀释剂再配制是产生具有约5.0-6.5pH范围的溶液。
Description
技术领域
领域：本发明通常涉及药物制剂领域。更具体地说，本发明涉及稳定的、具有治疗活性、能够选择性激活T细胞(PHA-胚细胞)的白介素-2制剂并且非常优选包括表明自然杀伤(“NK”)细胞激活减少的IL-2突变蛋白质。这种稳定的具有优选特性的组合物包括下文所述的IL-2变体。
背景技术
背景：如1999年11月25日公开的相关申请PCT/US99/10643所述，白细胞介素2(IL-2)是强的免疫刺激剂，激活免疫系统的多种细胞，包括T细胞、B细胞和单核细胞。IL-2也是T细胞的强的关键的生长因子。依据这些活性的效能来试验IL-2治疗癌症的能力。人IL-2是FDA批准的用于治疗转移的肾癌和转移的黑素瘤。IL-2在合适患者中的使用因其与IL-2治疗相关的严重的毒性而受到限制，估计实际上最好只有20％的合适患者接受治疗。与IL-2治疗相关的毒性包括严重的发烧、恶心、呕吐、血管渗漏和严重的低血压。尽管有这些毒性，IL-2对于其批准的适应症是有效的。IL-2降低毒性的变体是WO99/60128申请的主题。
可获得有关稳定IL-2和其他治疗性蛋白质制剂的有意义的信息。目前批准的人IL-2制剂(IL-2，Chiron Corporation)是冷冻干燥制剂，其包括甘露糖醇、十二烷基硫酸钠(SDS)和磷酸盐缓冲液。在下列参考文献中描述了所配制的其他治疗性蛋白质包括IL-2。
Fernandes等人，1986，由微生物产生的白介素-2的药物组合物(美国专利4,604,377)中描述了冷冻干燥的制剂，它包含稳定剂(甘露糖醇)和每毫克IL-2约100到250μg的增溶剂如十二烷基硫酸钠或者脱氧胆酸硫酸钠。相信在该参考文献中描述了目前可获得的IL-2产品的制剂。
Patel，1994，蛋白质制剂的稳定(美国专利5358708)中描述了通过与蛋氨酸、组氨酸或其混合物混合来延长储存期的干扰素、粒细胞-巨噬细胞集落刺激因子或白介素的水性制剂。尽管以几种白介素包括白介素-2为参考，但在用IL-4进行研究中，专利权所有人发现，在所使用的稳定剂测试条件下，组氨酸稳定剂的有效性不如甲硫氨酸。
Shaked等人，1991，重组白介素-2的药物组合物及其制备方法(美国专利5,037,644)中描述了冻干或液体形式的制剂。所述制剂的赋形剂包括非离子型高分子去垢剂如吐温X405、吐温X305、PEG(4000)单硬脂酸酯，浓度大约为0.001％-5％的Tween 80和Tween 20，膨胀剂/稳定剂如蔗糖、果糖、右旋糖、麦芽糖、葡萄糖、右旋糖酐、甘露糖醇、山梨糖醇、肌醇、半乳糖醇、木糖醇、乳糖、海藻糖、人血清白蛋白和牛血清白蛋白，和浓度大约为10mM-50mM，pH范围大约为3-7的缓冲剂如甘氨酸、枸橼酸盐或磷酸盐。多元醇糖膨胀剂的浓度(wt/vol)浓度范围大约为0.025％-10％。
Roskam等人，1995，包含糖基化白介素-2的药物(美国专利5,417,970)中描述了pH6.5、包含水解明胶(或人血清白蛋白)和丙氨酸的冻干制剂。
Hora等人，1992，包含生理学上相容性稳定剂的白介素-2的药物制剂(美国专利5,078,997)中描述了液体或冻干形式的制剂。制剂可包含一种稳定剂或稳定剂的组合物，所述稳定剂如精氨酸、肉毒碱、甜菜碱、吡哆醇、聚乙烯吡咯烷酮、癸酸盐、糖、糖醇、血清白蛋白和pH为5.0-8.5的枸橼酸盐缓冲液。稳定剂的浓度如下：精氨酸为0.2-3.0％(w/v)、肉毒碱为0.2-3.0％(w/v)、蔗糖为2-6％(w/v)并且枸橼酸盐为0.01-0.3M。
Yasushi等人，1987，白介素-2和白蛋白的稳定的组合物(美国专利4,645,830)中描述了pH3-5.5、包含人血清白蛋白(0.1-50mg/ml)且包含或不包含还原性赋形剂如谷胱甘肽、硫辛酸、N-乙酰基半胱氨酸或抗坏血酸(浓度0.05-20mg/ml)的稳定的水性制剂。白蛋白制剂可包含单氨基脂肪族氨基酸、环状氨基酸、单糖、糖醇或单氨基脂肪族氨基酸(浓度为5-50mg/ml)。
Lee等人，1989，在低离子强度介质氯化钠和/或氯化钾、赖氨酸盐酸盐和组氨酸中的药用血浆蛋白质制剂(美国专利4,877,608)中描述了在包含氯化钠、氯化钾或其混合物；赖氨酸盐酸盐；和组氨酸作为缓冲剂的低离子强度介质中因子VIII和其他血浆蛋白质的稳定的制剂。
Nayar，1998，稳定的低含糖量无白蛋白重组因子VIII制剂(美国专利5,763,401和5,874,408)中描述了稳定的无白蛋白含甘氨酸、组氨酸、蔗糖和NaCL的FVIII制剂。
为了尝试发现稳定的IL-2制剂(特别是WO 99/60128中优选的IL-2突变蛋白质(N88R))，我们现已找到具有生物活性的和有用的人IL-2的非常稳定的和可药用的制剂。如下所述，我们的发现基于我们相信影响稳定性的基本机理。
附图简述
图1是说明IL-2水溶液最佳pH范围的图。
图2比较了组氨酸与醋酸盐和枸橼酸盐对IL-2凝聚的稳定作用，所述凝聚是以1℃/min的速度将蛋白质溶液从25℃加热到95℃引起的。
发明概述
本发明是用组氨酸稳定的IL-2或其变体(突变蛋白质)的药物组合物或制剂。优选地，所述组合物包含在低离子强度(例如＜0.1)溶液中产生的混合物并且包含其他稳定剂如糖和氨基酸，优选蔗糖和甘氨酸。所述制剂可包含0-0.9wt.％NaCl。组合物不合白蛋白并且冻干形式的制剂可以用水快速再配制。组合物在生理上适宜的pH条件下，优选在pH大约为5.0-6.5的范围内，并且不使用表面活性剂如十二烷基硫酸钠的情况下是稳定的。重新配制的溶液近等渗的并且可通过皮下和静脉内给予。在非常优选的实施方案中，组合物中的IL-2是具有单一的氨基酸取代的突变蛋白质，优选WO 99/60128中所描述的N88R变体。
非常优选的组合物具有1-5mg/ml的蛋白质浓度并且包含下列水溶液形式(以wt/wt方式)：
IL-2 0.1-0.5wt％
组氨酸 0.08-1.6wt％
NaCl 0-0.9wt％
蔗糖 1-10wt％
甘氨酸 0-3wt％
pH为 5-6.5。
下文详细描述我们的制剂及其是如何发现的。
具体实施方案
本文所使用的术语IL-2包括有活性的野生型IL-2及其具有生物活性的变体或突变蛋白质如WO 99/60128中所描述的那些。在下列实施例中，我们使用称作IL-2(N88R)的IL-2，它是在氨基酸88位上的天门冬氨酸(N)突变为精氨酸(R)的人IL-2重组突变蛋白质。该突变蛋白质是由中国仓鼠卵巢(CHO)细胞表达的并且包括糖基化形式和非糖基化形式的混合物。它描述于以上所引用的作为相关申请的WO 99/60128中。
本发明的目的是识别优选的IL-2突变蛋白质冻干剂型的需要，所述剂型不含白蛋白并且具有适宜的稳定性。既然IL-2的生物测定是测定稳定性灵敏的方法，那么我们通过使用反向HPLC测定可溶性IL-2的量作为指示稳定性的测定。本文所使用的术语“稳定的”或“稳定”是指在40℃下贮存4个月后，通过反向HPLC测定的可溶性IL-2量的减少不低于原可溶性IL-2量的90％(见下表3和4)。所使用的其他指示稳定性的测定包括聚集指数，通过UV/VIS分光光度法测定聚集和通过尺寸排阻HPLC法测定可溶性聚集体。除了稳定的产品外，快速再配制(少于1分钟)的冻干物是非常优选的。最后，需要具有适宜的稳定性并且可以很容易地在生产冷冻干燥器中干燥的冻干剂型制剂。
在预制剂研究期间进行的水稳定性研究表明IL-2(N88R)在液态迅速聚集并且所述聚集是pH依赖性的。进行了两种预制剂稳定性研究：IL-2(N88R)的pH图和不同缓冲剂存在下的稳定性。这些研究的目的是识别适用于IL-2(N88R)的适宜的pH范围和适用于水稳定(减少聚集的可能性)的适宜的缓冲剂。在绘制pH图期间，制备具有不同的pH值的IL-2溶液并且在加速聚集的温度(40℃)下贮藏。在不同的时间间隔下分析样品并计算聚集速度。如图1所示，低聚集速度的最佳pH范围为pH 5.0-6.5。组氨酸、醋酸盐和枸橼酸盐是在该pH范围内IL-2(N88R)的药用缓冲剂并且在浓度为20mM、包含1mg/ml IL-2(N88R)和150mM(0.9wt％)NaCl的IL-2(N88R)溶液中进行评价。将这些样品以每分钟1℃的速度从25℃加热至95℃并且通过UV分光光度计在350nm监测沉淀。如图2所示，令我们意外的是，在pH 5.5时，组氨酸稳定IL-2(N88R)的作用明显地超过其他缓冲剂，这一点可通过沉淀的起始温度增加表明。在枸橼酸盐、醋酸盐和组氨酸存在下，起始温度分别为62℃、64℃和70℃。这些研究表明，对于含水条件下的IL-2来说，组氨酸不仅可用作缓冲剂，而且可用作稳定剂。
为了开发冻干剂型，研究了熟悉本领域的那些人所使用的其他赋形剂。这些赋形剂包括膨胀剂和低温保护剂如甘氨酸、蔗糖和甘露糖醇。既然聚集是在水稳定性研究时一种分子不稳定机理，那么也评价两种作为IL-2(N88R)稳定剂的表面活性剂。这些研究结果概述于下列实施例中。意外地，结果显示组氨酸对IL-2的选择性稳定作用优于其他赋形剂，如枸橼酸盐。
实施例1
在40℃下测定变体pH值的液体IL-2(N88R)的稳定性。结果显示在ph5.0-6.5时IL-2(N88R)的沉淀速度最小(图1)。因此，pH5.5选作液态制剂的最佳pH并且用于制备冻干制剂。
实施例2
我们测定不同的缓冲剂对IL-2(N88R)稳定性可能的作用。所测定的缓冲剂包括枸橼酸盐、醋酸盐和组氨酸。所使用的缓冲剂为20mM、包含1mg/ml IL-2(N88R)和150mM(0.9wt％)NaCl的IL-2(N88R)溶液。将这些稳定样品以每分钟1℃的速度从25℃加热至95℃同时通过UV/VIS分光光度计监测。与醋酸盐和枸橼酸盐相比，组氨酸通过增加沉淀温度和降低沉淀速度明显地稳定IL-2(N88R)(图2)。表1显示在这三种缓冲剂存在下IL-2(N88R)的起始沉淀温度。起始沉淀温度任意地定义为350nm下的光密度(OD350)达到某种水平(在OD350的情况下为0.2和1.0)时的温度。在组氨酸存在下，IL-2(N88R)的沉淀温度比其他缓冲剂存在下高几度。该实例证明，组氨酸除了用作液态IL-2(N88R)的缓冲剂外，可能是特异性稳定剂。
表1：IL-2(N88R)在不同缓冲剂中的沉淀温度(OD350＝0.2和OD350＝1.0)
| 缓冲剂 | 沉淀温度℃ (当OD<sub>350</sub>＝0.2时) | 沉淀温度℃ (当OD<sub>350</sub>＝1.0时) |
| 枸橼酸盐 | 62℃ | 64℃ |
| 醋酸盐 | 64℃ | 66℃ |
| 组氨酸 | 70℃ | 76℃ |
实施例3
为了进一步评价组氨酸的稳定作用，由不同的水溶液制剂(见表2)制备冻干的IL-2(N88R)。这些制剂的大多数都含有2wt％作为膨胀剂的甘氨酸和1wt％作为稳定剂的蔗糖。在作为对照的
(一种IL-2的商业化产品)制剂中使用5wt％的甘露糖醇。评价0.1wt％两种表面活性剂(吐温80和Pluronic F68)对表面吸附和凝聚的防止作用。所有制剂都含有组氨酸或者作为缓冲剂的枸橼酸以使pH调节到5.5。包含枸橼酸来区别组氨酸的稳定作用与pH5.5时枸橼酸的作用。在40℃温度下贮存这些冻干制剂并通过各种分析方法来进行分析，包括UV/VIS分光光度测定法、SEC-HPLC和RP-HPLC。
表3显示通过下列方法评价的各种制剂的稳定性：通过UV/VIS分光光度测定法测定凝聚、通过大小排阻HPLC(SEC-HPLC)测定可溶解凝聚物的量并通过反向HPLC(RP-HPLC)测定蛋白质的回收百分数。样品在冻干后和在40℃加速贮存温度下贮存4个月后进行分析。冻干过程不改变冻干前IL-2(N88R)的凝聚指数，暗示IL-2(N88R)在蛋白质凝聚/沉淀方面耐受冻干过程。在40℃温度下贮存4个月后，观察到含有枸橼酸(B)、Pluronic F68(D)或甘露糖醇(E)制剂的净凝聚指数显著增加。这些结果表明在贮存期间枸橼酸或甘露糖醇、吐温80或Pluronic F68不对固体状态的IL-2(N88R)凝聚提供任何保护作用。制剂A和F的凝聚指数不显示明显的改变，暗示具有1或5mg/ml IL-2(N88R)的组氨酸、甘氨酸和蔗糖产生稳定的产品。
然而，发现在含有吐温80的制剂甚至在冻干前就有明显量的可溶性凝聚物(表3)，表明吐温80尽管可抑制不溶性凝聚物的形成，但促进可溶性IL-2(N88R)凝聚物形成。因为Pluronic F68(制剂D)也引起可溶性凝聚物明显形成，表面活性剂与IL-2(N88R)不相容。只有含2％甘氨酸、1％蔗糖和20M(0.31wt％)组氨酸(A，F)的制剂未显示所述冻干制剂在40℃下贮藏4个月后形成可检测到的可溶性聚集体，再次暗示IL-2(N88R)可由组氨酸稳定。
通过RP-HPLC测定冻干和贮藏后IL-2(N88R)的总回收率(表3)。除了含甘露糖醇的制剂外，所有制剂冻干后IL-2(N88R)的回收率大于大约96％。这些制剂在40℃下贮藏4个月后，在含2％甘氨酸、1％蔗糖和20mM(0.31wt％)组氨酸并且分别含1和5mg/ml IL-2(N88R)的制剂A和F中IL-2(N88R)的回收率大约为92％。这些数据(制剂A和F中IL-2(N88R)的回收率大于90％)再次暗示组氨酸稳定IL-2(N88R)，而表面活性剂不能稳定蛋白质。
表2：冻干IL-2(N88R)制剂的组成
| 制剂ID | A | B | C | D | E | F |
| IL-2(N88R)，mg/ml | 1 | 1 | 1 | 1 | 1 | 5 |
| 甘氨酸，wt％(w/w) | 2 | 2 | 2 | 2 | 0 | 2 |
| 蔗糖，wt％(w/w) | 1 | 1 | 1 | 1 | 0 | 1 |
| 甘露糖醇，wt％(w/w) | 0 | 0 | 0 | 0 | 5 | 0 |
| 枸橼酸钠，wt％(w/w) | 0 | 0.6 | 0 | 0 | 0 | 0 |
| 组氨酸wt％(w/w) | 0.31 | 0 | 0.31 | 0.31 | 0.31 | 0.31 |
| 吐温80，wt％(w/w) | 0 | 0 | 0.1 | 0 | 0 | 0 |
| Pluronic F68，wt％(w/w) | 0 | 0 | 0 | 0.1 | 0 | 0 |
| pH | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 |
[0043]表3：IL-2(N88R)在冻干过程中和冻干制剂在40℃下贮藏4个月期间的稳定性
| 制剂ID | A | B | C | D | E | F |
| 聚集指标(％) | ||||||
| 冻干前 | 4.4 | 5.2 | 1.5 | 4.7 | 5.6 | 1.6 |
| 冻干后 | 5.2 | 5.1 | 0.7 | 1.5 | 3.8 | 1.6 |
| 在40℃下4个月 | 2.9 | 13.9 | 2.4 | 14.3 | 20.8 | 3.3 |
| 通过SEC-HPLC测定可溶性聚 集体(％) | ||||||
| 冻干前 | ND<sup>a</sup> | ND | 4.4％ | ND | ND | NA<sup>b</sup> |
| 冻干后 | ND | ND | 7.2％ | ND | ND | NA |
| 在40℃下4个月 | ND | 4.2％ | 32.9％ | 15.9％ | 12.2％ | NA |
| 通过RP-HPLC测定回收率(％) | ||||||
| 冻干前 | 100 | 100 | 100 | 100 | 100 | 100 |
| 冻干后 | 96.5 | 95.7 | 96.4 | 99.4 | 91.7 | 97.8 |
| 在40℃下4个月 | 92.5 | 82.9 | 71.9 | 84.3 | 79.1 | 91.7 |
aND＝不可检测的
bNA＝不可获得的
实施例4
也将与制剂A具有相同组成的野生型IL-2的水溶液制剂冻干。比较野生型IL-2与IL-2(N88R)在40℃下的稳定性(表4)。
表4；野生型IL-2(N88R)制剂在冻干过程中和冻干制剂在储存期间的稳定性
| 条件 | 聚集指标(％) | 可溶性聚集体(％) | 回收率(％) |
| 冻干前 | 4.2 | ND<sup>a</sup> | 100 |
| 冻干后 | 5.1 | ND | 93.6 |
| 在40℃下4个月 | 4.3 | ND | 93.0 |
aND＝不可检测的
讨论
人IL-2具有133个氨基酸，它们形成六种螺旋结构(A-F)。这些螺旋中的四种形成称作四-螺旋束的基元。Cys58和cys105之间的分子内二硫键位于螺旋间延长的环状结构上。游离cys125位于结合氨基酸117-133的螺旋F上。
对组氨酸是IL-2特异性稳定剂的惊奇发现暗示组氨酸可能与IL-2以特异的方式相互作用，由此稳定水溶液状态和冻干状态的分子。IL-2不稳定的主要机制之一是凝聚，这是由于硫醇-二硫化物的交换反应形成低聚物所致。因此，一种假设是事实上组氨酸可抑制或减少IL-2中硫醇-二硫化物的交换反应。由于野生型IL-2、IL-2(N88R)和可能的其他IL-2变体分子具有一个二硫键和游离的半胱氨酸(cys125)，cys125上的游离-SH基团易于通过硫醇/二硫化物交换途径与二硫键进行反应，由此导致凝聚/沉淀发生。
Bulaj等人最近描述了硫醇/二硫化物交换机制(多肽中半胱氨酸硫醇的离子化-反应活性的关系，Biochemistry 1998，6，23；37(25)：8965-72)。在模型肽和蛋白质的研究中，显示反应速率对静电力以及蛋白质的二级结构是敏感的。硫醇中硫原子周围的电子分布可受改变硫醇pKa的附近电荷和整个键诱导作用存在的影响。例如，硫醇/二硫化物中硫醇反应活性增加归因于其pKa因附近正电荷或者来自附近α螺旋结构的肽偶极分布的降低。相反地，巯基附近的负电荷可使硫醇的pKa升高并导致硫醇/二硫化物反应速率的降低。所提出的机制也得到其他研究的支持，已经表明通过邻接正电荷残基稳定蛋白酪氨酸磷酸酯酶(Zhang和Dixon，1993Activesite labeling ofthe Yersinia protein tyrosine phosphatase：the determination ofthe pKa of the active site cysteine and the function of theconserved histidine 402.Biochemistry 1993Sep 14；32(36)：9340-5)和蛋白二硫化物异构酶(Kortemme等人，Electrostatic interactions in the active site of the N-terminalthioredoxin-like domain of protein disulfide isomerase.Biochemistry 1996Nov 19；35(46)：14503-11)中高反应活性的硫醇盐离子。
基于硫醇/二硫化物交换反应的这些特性，可以预想一种稳定机制，那就是组氨酸在IL-2(N88R)的稳定中起动力学或者热力学作用。在二硫化物桥Cys58-Cys105附近有5个谷氨酸残基(57，60，61，62和106)。组氨酸和这些负电荷残基的特异性结合可导致硫醇/二硫化物交换反应的动力学屏障或者二硫化物桥附近组氨酸的结合可在热力学上将IL-2分子稳定为不易于进行凝聚反应的构型。另外，His-Glu离子的相互作用也可产生立体上的障碍，这将进一步使硫醇/二硫化物交换中决定速率的步骤难以进行，其中在三个参与的硫原子之间形成中间体过渡状态。组氨酸易于接近谷氨酸残基是非常可能的，因为二硫化物位于蛋白质表面的延长的环结构上。
给出了以上实施例，预计本领域技术人员可对本文所公开的发明进行改变。因此，希望上述实施例仅仅是说明性的并且本文公开的发明将仅通过下列权利要求来限定。
Claims (6)
1.一种稳定的冻干药物组合物，包含每毫升1至5毫克的人白介素-2突变蛋白质，以及包含组氨酸、甘氨酸和蔗糖的稳定混合物，其中所述人白介素-2突变蛋白质由带有一个替换的野生型人IL-2的氨基酸序列组成，所述替换是在野生型人IL-2的第88位的天冬酰胺被精氨酸替代。
2.权利要求1的稳定的冻干药物组合物，其中以重量计，组氨酸的量为0.08-1.6％。
3.权利要求1的稳定的冻干药物组合物，其中以重量计，甘氨酸的量为0-3％。
4.权利要求1的稳定的冻干药物组合物，其中以重量计，蔗糖的量为1-10％。
5.权利要求1的稳定的冻干药物组合物，其为含水形式，具有5.0-6.5的pH范围。
6.一种稳定的冻干药物组合物，在用水再配制后，它含有下列组分：
1.0-5.0mg/ml的人IL-2突变蛋白质，其由带有一个替换的野生型人IL-2的氨基酸序列组成，所述替换是在野生型人IL-2的第88位的天冬酰胺被精氨酸替代
组氨酸 0.08-1.6wt％
NaCl 0-0.9wt％
蔗糖 1-10wt％和
甘氨酸 0-3wt％
pH 5-6.5。
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| JP2023510115A (ja) | 2019-12-20 | 2023-03-13 | リジェネロン・ファーマシューティカルズ・インコーポレイテッド | 新規ｉｌ２アゴニストおよびそれらの使用方法 |
| EP4087865A2 (en) | 2020-01-10 | 2022-11-16 | Bright Peak Therapeutics AG | Modified il-2 polypeptides and uses thereof |
| JP7303391B2 (ja) | 2020-01-14 | 2023-07-04 | シンセカイン インコーポレイテッド | バイアス型ｉｌ２ムテイン、方法、および組成物 |
| WO2021168079A1 (en) | 2020-02-21 | 2021-08-26 | Pandion Operations, Inc. | Tissue targeted immunotolerance with a cd39 effector |
| BR112022022826A2 (pt) | 2020-06-03 | 2022-12-13 | Ascendis Pharma Oncology Div A/S | Sequências de il-2 e usos das mesmas |
| TWI815194B (zh) | 2020-10-22 | 2023-09-11 | 美商基利科學股份有限公司 | 介白素2-Fc融合蛋白及使用方法 |
| US20230398183A1 (en) * | 2020-11-13 | 2023-12-14 | Jiangsu Hengrui Pharmaceuticals Co., Ltd. | Pharmaceutical composition comprising human interleukin 2 variant or derivative thereof and use thereof |
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| JPH0645551B2 (ja) * | 1986-01-07 | 1994-06-15 | 塩野義製薬株式会社 | インタ−ロイキン−２組成物 |
| JPH0229016A (ja) * | 1989-06-09 | 1990-01-31 | Alpine Electron Inc | イコライザ装置 |
| US6267958B1 (en) * | 1995-07-27 | 2001-07-31 | Genentech, Inc. | Protein formulation |
| US5763401A (en) | 1996-07-12 | 1998-06-09 | Bayer Corporation | Stabilized albumin-free recombinant factor VIII preparation having a low sugar content |
| AU8985898A (en) * | 1997-07-31 | 1999-02-22 | Rhodia Chimie | Cosmetic composition comprising a functionalised polyorganosiloxane |
| DZ2788A1 (fr) * | 1998-05-15 | 2003-12-01 | Bayer Ag | Agonistes et antagonistes selectifs à IL-2. |
| US6689353B1 (en) * | 2000-06-28 | 2004-02-10 | Bayer Pharmaceuticals Corporation | Stabilized interleukin 2 |
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| US5917021A (en) * | 1995-04-07 | 1999-06-29 | Enzon, Inc. | Stabilized monomeric protein compositions |
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