CN114057863A - 一种甲状旁腺激素相关肽类似物及其应用 - Google Patents
一种甲状旁腺激素相关肽类似物及其应用 Download PDFInfo
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- CN114057863A CN114057863A CN202010780705.5A CN202010780705A CN114057863A CN 114057863 A CN114057863 A CN 114057863A CN 202010780705 A CN202010780705 A CN 202010780705A CN 114057863 A CN114057863 A CN 114057863A
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Abstract
本发明提供一种甲状旁腺激素相关肽类似物及其应用。该甲状旁腺激素相关肽类似物为通式Ⅰ所示的化合物或其盐或溶剂合物。该甲状旁腺激素相关肽类似物具有良好的稳定性,稳定性优于特立帕肽且不劣于阿巴帕肽;其对于PTHR1具有很高的激动活性,激动活性显著高于特立帕肽和阿巴帕肽,在做为预防和/或治疗骨骼疾病及其相关并发症药物时具有药物活性优和成骨效果好的特点。
Description
技术领域
本发明涉及医药领域,更具体地讲,本发明涉及一种甲状旁腺激素相关肽类似物及其应用。
背景技术
目前全球罹患骨质疏松症的患者数量超过2亿,平均每3秒便会有一起因骨质疏松引发的骨折案例,而髋部骨折患者在骨折后1年内的死亡率高达20%。尽管此数字已如此明显,但随着全球人口的老龄化,此数字依然会不断增加。因此越来越迫切需要寻找安全有效的药物治疗这些因骨质减少引起的疾病。
经过多年的探索研究,人们发现甲状旁腺激素(PTH)和甲状旁腺激素相关肽(PTHrP)共享很多生物学作用,包括都可结合甲状旁腺受体1(PTHR1),结合后可调节钙、磷代谢及骨转换,这与骨质减少有着密切的关联。两者的活性片段有PTH(1-34),PTH(1-84),PTHrP(1-34),PTHrP(1-36)等,人们在此基础上开发出了一系列PTH类似物和PTHrP类似物的药物。
hPTH(1-34),氨基酸序列为:
Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly-Lys-His-Leu-Asn-Ser-Met-Glu-Arg-Val-Glu-Trp-Leu-Arg-Lys-Lys-Leu-Gln-Asp-Val-His-Asn-Phe-OH(SEQ IDNO:1)。
hPTHrP(1-34),氨基酸序列为:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-Lys-Gly-Lys-Ser-I1e-Gln-Asp-Leu-Arg-Arg-Arg-Phe-Phe-Leu-His-His-Leu-Ile-Ala-Glu-Ile-His-Thr-Ala-NH2(SEQID NO:2)。
然而,特立帕肽化学稳定性差,其溶液易降解,药物主要以氧化,脱酰胺和肽键裂解三种形式降解,其中最突出的是Met8和Met18残基的氧化,Asn10、Asn16和Asn 33处的脱酰胺作用导致肽键在Asn16,Asn33,Asp30处裂解(Ruchi Kothari et al.Modes ofDegradation and Impurity Characterization in rhPTH(1-34)during StabilityStudies.2011,65(4):348-62.)。上述原因导致其液体注射剂必须保存在2-8℃冰箱中,开封使用后也要2-8℃保存,不仅对储存和运输条件要求较高,也不利于病人出行使用。
为了克服PTH(1-34)易降解的缺点,首选的甲状旁腺激素类似物是由至少一个氨基酸被替换的,被替换到甲状旁腺激素序列的以下一个或多个位置:8-11、13、16-19、21、22、29至34,特别是8-11、16-19、33和/或34。例如Sandoz公司开发了一种新颖性的PTH(1-34)类似物SDZ PTS 893,其结构式是[Leu8,Asp10,Lys11,Ala16,Gln18,Thr33,Ala34]-hPTH-(1–34),增强了溶液中的化学稳定性和抗氧化性,在肾脏细胞的结合亲和力,刺激腺苷酸环化酶,增加细胞内的钙等方面是自然片段的2到5倍(R.Gamse et al.SDZ PTS 893:Pharmacological Profile of A Highly Potent Novel PTH Analog.ASBMR 19th AnnualMeeting:F366)。在去卵巢大鼠模型中对比了PTH(1-34),PTHrP(1-36)和SDZ PTS 893的治疗效果,以骨合成的能力排序为SDZ PTS 893>PTH(1-34)>PTHrP(1-36),但SDZ PTS 893组大鼠的副作用也是最大的,这些大鼠出现中度高钙血症和明显的肾钙积累,死亡率为13%(A.F.Stewart et al.Six-Month Daily Administration of Parathyroid Hormone andParathyroid Hormone-Related Protein Peptides to Adult Ovariectomized RatsMarkedly Enhances Bone Mass and Biomechanical Properties:A Comparison ofHuman Parathyroid Hormone 1-34,Parathyroid Hormone-Related Protein 1-36,andSDZ-Parathyroid Hormone 893.Journal of Bone and Mineral Research:the officialjournal of the American Society for Bone and Mineral Research.2000,15(8):1517-25.)。遗憾的是,到目前为止此类药物依然没有相关上市的信息。
Radius Health公司开发的阿巴帕肽([Glu22'25,Leu23'28'31,Aib29,Lys26'30]hPTHrP(l-34)NH2),氨基酸序列为:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-Lys-Gly-Lys-Ser-I1e-Gln-Asp-Leu-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Aib-Lys-Leu-His-Thr-Ala-NH2(SEQID NO:3),其注射液于2017年4月28日被FDA批准上市,是以促骨形成来治疗骨质疏松症的PTHrP类似物药物,商品名为
阿巴帕肽在hPTHrP(l-34)的基础上修改了8处氨基酸,增加药物活性的同时兼具优良的溶液稳定性,其液体注射剂在未使用前需2-8℃运输保存,开封使用后可在20-25℃保存30天,大大降低了保存的要求,患者不论居家使用或者外出携带使用都很方便,但其29位氨基酸是非天然氨基酸Aib,Radius Health公司使用SPSS合成,生产成本相比生物合成较高。
罗氏开发的一种新颖性的全天然氨基酸的PTHrP类似物RS66271即[Glu22'25'29,Leu23'28'31,Lys26'30]hPTHrP(l-34)NH2,50μg和100μg剂量在6个月的治疗周期中分别增加了绝经后骨质疏松妇女2.2%和8.8%的腰椎BMD,但因出现了皮质骨的丢失停止了进一步的开发(John Fox,Developments in parathyroid hormone and related peptides asbone-formation agents.Current Opinion in Pharmacology.2002,2:338–344.)。
特立帕肽和阿巴帕肽两者对骨骼的作用依赖于***暴露模式,以脉冲方式给药,需每日皮下注射,病人的顺应性和依从性差。由于在大鼠的致癌性研究中显示,使用两者可使骨肉瘤(一种恶性肿瘤)的发病率增加,故两者的治疗周期最长为24个月,病人终身仅可接受一次为期24个月的治疗。
所以目前急需有溶液常温稳定性优,方便患者使用携带,生产成本低廉,适合大规模生产,在有限的治疗时间内药物活性更好的PTHrP类似物。
发明内容
本发明的首要目的在于克服现有技术的缺点与不足,提供一种甲状旁腺激素相关肽(PTHrP)类似物。所述的甲状旁腺激素相关肽(PTHrP)类似物的多肽序列全部由天然氨基酸组成,可通过生物发酵生产,生产成本低廉,且所述甲状旁腺激素相关肽(PTHrP)类似物具有溶液稳定性,血浆稳定性和物理稳定性优良的特点。
本发明的第二个目的在于提供上述甲状旁腺激素相关肽(PTHrP)类似物在制备用于预防和/或治疗骨骼疾病及其相关并发症药物中的应用,且所述甲状旁腺激素相关肽(PTHrP)类似物具有药物活性优和成骨效果好的特点。
本发明的目的通过下述技术方案实现:
一种甲状旁腺激素相关肽(PTHrP)类似物,其是具有如通式Ⅰ所示的化合物或其盐或溶剂合物:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-X11-Gly-Lys-Ser-I1e-Ala-Asp-Ala-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-X29-Lys-Leu-His-Thr-Ala-NH2(通式Ⅰ)
其中,
X11选自Arg、Lys或His;
X29选自Leu、Ile、Val或Phe。
在一些实施例中,所提供的PTHrP类似物选自氨基酸序列如SEQ ID NO.4所示的化合物1、氨基酸序列如SEQ ID NO.5所示的化合物2、氨基酸序列如SEQ ID NO.6所示的化合物3或氨基酸序列如SEQ ID NO.7所示的化合物4:
化合物1:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-Arg-Gly-Lys-Ser-I1e-Ala-Asp-Ala-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Leu-Lys-Leu-His-Thr-Ala-NH2(SEQID NO.4);
化合物2:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-Arg-Gly-Lys-Ser-I1e-Ala-Asp-Ala-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Ile-Lys-Leu-His-Thr-Ala-NH2(SEQID NO.5);
化合物3:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-Arg-Gly-Lys-Ser-I1e-Ala-Asp-Ala-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Val-Lys-Leu-His-Thr-Ala-NH2(SEQID NO.6);
化合物4:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-Arg-Gly-Lys-Ser-I1e-Ala-Asp-Ala-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Phe-Lys-Leu-His-Thr-Ala-NH2(SEQID NO.7)。
所述的甲状旁腺激素相关肽(PTHrP)类似物,可通过包括合成法、重组DNA生物技术在内的方法制备得到。
所述的甲状旁腺激素相关肽(PTHrP)类似物在制备用于预防和/或治疗骨骼疾病及其相关并发症药物中的应用。
所述的预防和/或治疗骨骼疾病及其相关并发症药物包括预防和/或治疗骨质疏松症的药物组合物。
所述的甲状旁腺激素相关肽(PTHrP)类似物对PTH受体1(PTHR1)具有激动活性,因此所述的甲状旁腺激素相关肽(PTHrP)类似物还可以作为PTHR1激动剂。所述的“激动活性”是指化合物可激动特定受体细胞产生cAMP,所使用细胞可为由本领域技术人员构建的过表达PTH受体1的人胚肾细胞(HEK 293)、COS-7细胞或CHO-K1细胞。所述受体激动活性可采用化合物激动受体细胞产生cAMP的EC50值作为衡量数值。EC50值是在特定的测定体系中达到化合物最大活性一半(50%活性)时所需要的药物浓度值。
一种用于预防和/或治疗骨质疏松症的药物组合物,由有效量化合物和药学上可接受的辅料组成;有效量化合物为上述甲状旁腺激素相关肽(PTHrP)类似物,或是上述甲状旁腺激素相关肽(PTHrP)类似物与其他治疗骨质疏松疾病的药物的组合。
所述的有效量化合物是指既能够产生疾病缓解或治疗作用,又不至于产生损害性作用的化合物量。有效量可由主治医师根据患者的疾病严重程度、年龄、性别、体重、一般健康状况等适宜地确定。所述的患者为哺乳动物;更优选为牛、马、山羊、绵羊、狗、黑猩猩、兔、小鼠、大鼠、猴、猪和人的至少一种;最优选为人。
所述的辅料包括但不限于稀释剂、赋形剂、润滑剂、粘合剂、填充剂、防腐剂、表面活性剂、着色剂、矫味剂、乳化剂、助悬剂、胶凝剂、崩解剂、pH调节剂和增溶剂中的至少一种。
所述的辅料用于辅助将上述药物组合物制成不同的剂型,包括但不限于注射剂或冻干粉、片剂、贴剂、丸剂、锭剂、软胶囊剂、硬胶囊剂、颗粒剂、散剂、溶液剂、混悬剂和糖浆剂;更优选为注射剂或冻干粉。
所述的药物组合物可装载于药物载体材料和/或药物递送装置中使用。
所述的药物载体材料优选为微囊、微球、纳米粒和脂质体中的至少一种。
所述的其他治疗骨质疏松疾病的药物包括但不限于双磷酸盐类、降钙素、***类、选择性***受体调节剂类、锶盐类、活性维生素D及其类似物和护骨强骨的中药类药物中的至少一种。
所述的药物组合物的给药方式包括但不限于口服给药、吸入给药和肠胃外给药中的至少一种或几种;所述的肠胃外给药优选通过腹膜内、肌肉内、动脉内、静脉内、皮下、皮内注射或透皮微针方式给药。
所述的药物组合物的给药频率可以根据实际情况进行选择;优选至少一天给药一次、一周给药一次或一个月给药一次。
本发明相对于现有技术具有如下的优点及效果:
(1)本发明所述化合物具有良好的稳定性(溶液稳定性,血浆稳定性和物理稳定性),其稳定性优于特立帕肽且不劣于阿巴帕肽。
(2)本发明所述化合物对于PTHR1具有很高的激动活性,激动活性显著高于特立帕肽和阿巴帕肽,其在作为预防和/或治疗骨骼疾病及其相关并发症药物时具有药物活性优和成骨效果好的特点。
(3)本发明所述化合物多肽序列全部由天然氨基酸组成,可以通过生物发酵生产,生产成本低廉。
具体实施方式
下面结合实施例对本发明作进一步详细的描述,但本发明的实施方式不限于此。
本发明所使用的缩写具体含义如下:
PTHR 1:甲状旁腺受体1
cAMP:环磷酸腺苷
HPLC-MS:高效液相色谱-质谱
HPLC:高效液相色谱
HEK-293:人胚肾细胞
CHO-K1:中国仓鼠卵巢细胞
PBS:磷酸盐缓冲溶液
FBS:胎牛血清
EC50:半数效应浓度
DMSO:二甲基亚砜
ThT:硫代黄素T
特立帕肽和阿巴帕肽均委托合肥国肽生物科技有限公司进行合成制备;德谷胰岛素购自诺和诺德有限公司。
实施例1:化合物合成
本发明所有化合物均委托合肥国肽生物科技有限公司通过化学合成方法进行合成,该公司所提供的合成方法过程如下。所描述的每一具体合成步骤可以采用不同的材料和方式组合,以合成多种对应的本发明所述化合物或其盐。所使用的试剂和原料为本领域普通技术人员容易获得。特别地,下面实施例仅用于说明本发明,不应以任何方式限制本发明的范围。
材料:
本发明所用材料及试剂均购买自商业化商品,整个合成过程所使用的保护氨基酸如下:Fmoc-Ser(tBu)-OH、Fmoc-Pro-OH、Fmoc-Ala-OH、Fmoc-Gly-OH、Fmoc-Asp(OtBu)-OH、Fmoc-Leu-OH、Fmoc-Trp(Boc)-OH、Fmoc-Gln(Trt)-OH、Fmoc-Val-OH、Fmoc-Phe-OH、Fmoc-Arg(Pbf)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Lys(Boc)-OH、Fmoc-Thr(tBu)-OH、Fmoc-His(Trt)-OH、Fmoc-Tyr(tBu)-OH、Fmoc-Ile-OH。
下面以化合物3(SEQ ID NO:6)为例,说明本发明化合物的合成制备方法。
方法:
(1)Rink氨基树脂前处理:称取1g干燥Rink氨基树脂,用DMF浸泡溶胀10min,抽去溶剂。
(2)脱除保护基Fmoc:往上述处理好的Rink氨基树脂中加入20%(v/v)哌啶/DMF溶液,搅拌反应20min。反应过程中使用茚三酮显色法监测反应程度,如果树脂产生颜色变化(变成紫色),表示Fmoc脱除成功。反应结束后过滤除去溶剂,向反应体系中加入DMF搅拌洗涤树脂1min,重复洗涤3次,得到脱除Fmoc保护基的氨基树脂。
(3)偶联反应(肽键形成):将配置好的相应Fmoc保护的氨基酸溶液加入到步骤(2)脱除Fmoc保护基的氨基树脂反应器中,然后加入DIC/DMF溶液,搅拌反应1.5h。反应过程中使用茚三酮显色法监测反应进行程度,如果树脂颜色无变化(仍为无色),说明偶联成功。反应完成后过滤除去溶剂,向反应体系中加入DMF搅拌洗涤树脂1min,重复洗涤3次。重复以上操作,依次加入相应的氨基酸溶液直至肽链合成完毕。最后一个氨基酸用Fmoc-Ala-OH进行偶联。化合物3主肽序列合成加入氨基酸偶联的顺序依次为Fmoc-Ala-OH、Fmoc-Thr(tBu)-OH、Fmoc-His(Trt)-OH、Fmoc-Leu-OH、Fmoc-Lys(Boc)-OH、Fmoc-Val-OH、Fmoc-Leu-OH(2×)、Fmoc-Lys(Boc)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Leu-OH(2×)、Fmoc-Glu(OtBu)-OH、Fmoc-Arg(Pbf)-OH(3×)、Fmoc-Ala-OH、Fmoc-Asp(OtBu)-OH、Fmoc-Ala-OH、Fmoc-Ile-OH、Fmoc-Ser(tBu)-OH、Fmoc-Lys(Boc)-OH、Fmoc-Gly-OH、Fmoc-Arg(Pbf)-OH、Fmoc-Asp(OtBu)-OH、Fmoc-His(Trt)-OH、Fmoc-Leu-OH(2×)、Fmoc-Gln(Trt)-OH、Fmoc-His(Trt)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Ser(tBu)-OH、Fmoc-Val-OH、Fmoc-Ala-OH。
(4)肽树脂后处理:加入切割试剂K进行切割树脂。过滤滤液,将滤液沉淀离心,得白色固体为目标化合物3粗品。
肽化合物粗品纯化:
将所得的肽粗品通过反相C18制备型色谱柱(岛津,Inertsil ODS 20×250mm 5μm)进行纯化,将样品纯化至纯度大于95%。以95%(v/v)缓冲液A(0.065%(v/v)TFA/水)和5%(v/v)缓冲液B(0.05%(v/v)TFA/乙腈)为起始洗脱液,以2%/min的速度逐渐增加缓冲液B的比例至65%,连续运行30min进行洗脱,收集目标肽组分。通过分析型HPLC/MS方法分析确证纯化的肽化合物。
基于以上合成方法,本发明合成了氨基酸序列如SEQ ID NO.4~7所示的多肽化合物1~4并进行表征(参见表1)。
表1.合成的肽化合物列表及分子量与纯度
实施例2:化合物稳定性测试
1.将特立帕肽,阿巴帕肽,化合物1-4使用乙酸钠缓冲液溶解(5mg/mL,由三水合乙酸钠配制)并用适量的乙酸调节pH,使溶液最终浓度都为2mg/mL,pH为5.1,用0.22μm无菌滤头过滤。分别吸取上述化合物溶液,通过HPLC(Shimadzu LC-20,Waters XBridge C18色谱柱,溶液A:0.1%(v/v)TFA-水溶液,溶液B:0.1%(v/v)TFA-ACN溶液;溶液A和溶液B中的溶剂分别为水和ACN)。分析15μL注射物的峰面积,此分析结果为该化合物稳定性测试的初始点(T0)。
将进行稳定性测试的化合物样品溶液置于25℃恒温箱中,密封避光保存6天。每天定时取样,该过程结束后,将样品溶液4500rpm离心10min,轻轻吸取上清溶液并通过HPLC分析10μL注射物的峰面积,此分析结果为该化合物稳定性测试的终点(T6)。
通过比较T0和各时间点测定化合物的目标峰面积及相关杂质峰面积,计算测定肽的剩余肽量。计算公式如下:
剩余肽(%)=(各时间点主峰面积/T0主峰面积)×100%;
通过比较测定肽化合物的肽剩余量来评估肽化合物的化学稳定性,测定结果参见表2。
表2肽化合物乙酸钠缓冲液稳定性分析结果
结果分析:从表中可以明显看出化合物1、化合物2、化合物3、化合物4在乙酸钠缓冲液中稳定性优于特立帕肽,与阿巴帕肽无差别。
2.实验前将冷冻血浆在37℃水浴中解冻,血浆以4000rpm离心5min,如果有血块,则去除血块。分别使用超纯水配制化合物溶液(12.5μM)和阳性对照药溴丙胺太林溶液(100μM),各取2μL化合物溶液和溴丙胺太林溶液加入98μL的空白血浆中37℃水浴锅孵育,在0、1、2、3、4、5、6、8、10h取样的样品中加入400μL终止液。将各样品4000rpm离心5min,取50μL上清液使用纯化水稀释至100μL,稀释后的样品800rpm震荡10min后使用HPLC-MS检测(ACQUITY UPLC,XBridge Protein BEH C4(蛋白分析)液相色谱柱,溶液A:0.1%(v/v)TFA-水溶液,溶液B:0.1%(v/v)TFA-ACN溶液)。
通过比较T0和各时间点测定化合物的目标峰面积及相关杂质峰面积,计算测定肽的剩余肽量。计算公式如下:
剩余肽(%)=(各时间点主峰面积/T0主峰面积)×100%;
通过比较测定肽化合物的肽剩余量来评估肽化合物的血浆稳定性,测定结果参见表3。
表3肽化合物人血浆稳定性分析结果
结果分析:从表中可以明显看出化合物1~4在人血浆稳定性试验中稳定性皆优于特立帕肽和阿巴帕肽。
3.将特立帕肽,阿巴帕肽,化合物1-4使用乙酸钠缓冲液溶解(5mg/mL,由三水合乙酸钠配制)并用适量的乙酸调节pH,使溶液最终浓度都为2mg/mL,pH为5.1,用0.22μm无菌滤头过滤。将1μM硫代黄素T(ThT)添加至样品,两者体积比例为ThT:样品=1:9。另取德谷胰岛素原研样品(按阿巴帕肽相同浓度和pH配制)按上述条件添加ThT。在96孔板中每孔加入100μL的样品,使用封板摸密封。在Cytation 5多功能酶标仪(Bio-Tek)中将温度调节至47℃,振幅为1mm,调节至807cpm的定轨道振摇下温育该板。利用通过440nm滤光片的激发以及测量485nm滤光片的发射,间隔20min进行荧光测量。
将测量点以Microsoft Excel格式保存以供进一步处理,并使用GraphPad Prism进行曲线绘制和拟合。可忽略在不存在原纤维时来自ThT的背景发射。
通过比较测定肽化合物的ThT原纤维形成的滞后时间来评估肽化合物的物理稳定性,测定结果参见表4。
表4肽化合物ThT原纤维形成实验分析结果
结果分析:从表中可以明显看出化合物1、化合物2、化合物3、化合物4在物理稳定性试验中稳定性与特立帕肽和阿巴帕肽相当,皆优于已上市的德谷胰岛素注射液,有着良好的物理稳定性。
实施例3:化合物对PTH受体1的活性测定
通过测定稳定过表达人PTH受体1的CHO-K1细胞(购买自南京金斯瑞生物科技股份有限公司;下同)的cAMP信号响应来确定肽化合物对相应受体的激动活性。细胞内cAMP含量使用Cisbio Corp.的试剂盒(cAMP Gs dynamic kit)基于HTRF(均相时间分辨荧光)技术测定。
将稳定过表达人PTH受体1的CHO-K1细胞(购买自南京金斯瑞生物科技股份有限公司)培养于含10%(v/v)的FBS和90%Ham's F-12K完全培养基中,待细胞长至80%~90%密度时,用0.25%(w/v)胰酶-EDTA消化,完全消化后将细胞团轻轻吹散成单个细胞,调节细胞密度为5.0×105个/mL。
取96孔板,分别向96孔板中每孔加入25μL细胞悬液(细胞密度为5.0×105个/mL)。待测化合物溶于1×PBS缓冲液中,从400nM按4倍逐级稀释,共配制8个浓度点的化合物溶液。利用移液枪将25μL配置好的化合物溶液分别加入96孔板对应细胞悬液中,吹打混合均匀,随后37℃孵育30min。药物孵育完成后各孔加入25μL试剂盒中的检测试剂,再室温孵育60min。将板置于Bio-Tek多功能酶标仪(Cytation5)中测定665nm/620nm处荧光读数。利用GraphPad Prism7.00作图软件制作化合物浓度-效应曲线并计算EC50值。
表5肽化合物的平均EC50值
结果分析:从表中可以明显看出化合物1、化合物2、化合物3、化合物4在受体激动活性试验中激动受体能力皆高于特立帕肽和阿巴帕肽。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
序列表
<110> 珠海联邦制药股份有限公司
<120> 一种甲状旁腺激素相关肽类似物及其应用
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<170> SIPOSequenceListing 1.0
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<211> 34
<212> PRT
<213> 人工序列(Artificial Sequence)
<223> hPTH(1-34)氨基酸序列
<400> 1
Ser Val Ser Glu Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn
1 5 10 15
Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His
20 25 30
Asn Phe
<210> 2
<211> 34
<212> PRT
<213> 人工序列(Artificial Sequence)
<223> h PTHrP(1-34)氨基酸序列
<400> 2
Ala Val Ser Glu His Gln Leu Leu His Asp Lys Gly Lys Ser Ile Gln
1 5 10 15
Asp Leu Arg Arg Arg Phe Phe Leu His His Leu Ile Ala Glu Ile His
20 25 30
Thr Ala
<210> 3
<211> 34
<212> PRT
<213> 人工序列(Artificial Sequence)
<223> 阿巴帕肽氨基酸序列
<221> ACT_SITE
<222> (29)..(29)
<223> Xaa=Aib
<400> 3
Ala Val Ser Glu His Gln Leu Leu His Asp Lys Gly Lys Ser Ile Gln
1 5 10 15
Asp Leu Arg Arg Arg Glu Leu Leu Glu Lys Leu Leu Xaa Lys Leu His
20 25 30
Thr Ala
<210> 4
<211> 34
<212> PRT
<213> 人工序列(Artificial Sequence)
<223> 化合物1氨基酸序列
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Ala Val Ser Glu His Gln Leu Leu His Asp Arg Gly Lys Ser Ile Ala
1 5 10 15
Asp Ala Arg Arg Arg Glu Leu Leu Glu Lys Leu Leu Leu Lys Leu His
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Thr Ala
<210> 5
<211> 34
<212> PRT
<213> 人工序列(Artificial Sequence)
<223> 化合物2氨基酸序列
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Ala Val Ser Glu His Gln Leu Leu His Asp Arg Gly Lys Ser Ile Ala
1 5 10 15
Asp Ala Arg Arg Arg Glu Leu Leu Glu Lys Leu Leu Ile Lys Leu His
20 25 30
Thr Ala
<210> 6
<211> 34
<212> PRT
<213> 人工序列(Artificial Sequence)
<223> 化合物3氨基酸序列
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Ala Val Ser Glu His Gln Leu Leu His Asp Arg Gly Lys Ser Ile Ala
1 5 10 15
Asp Ala Arg Arg Arg Glu Leu Leu Glu Lys Leu Leu Val Lys Leu His
20 25 30
Thr Ala
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<211> 34
<212> PRT
<213> 人工序列(Artificial Sequence)
<223> 化合物4氨基酸序列
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Ala Val Ser Glu His Gln Leu Leu His Asp Arg Gly Lys Ser Ile Ala
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Asp Ala Arg Arg Arg Glu Leu Leu Glu Lys Leu Leu Phe Lys Leu His
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Thr Ala
Claims (10)
1.一种甲状旁腺激素相关肽类似物,其特征在于,其是具有如通式Ⅰ所示的化合物或其盐或溶剂合物:
Ala-Va1-Ser-Glu-His-Gln-Leu-Leu-His-Asp-X11-Gly-Lys-Ser-I1e-Ala-Asp-Ala-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-X29-Lys-Leu-His-Thr-Ala-NH2 通式Ⅰ;
其中,
X11选自Arg、Lys或His;
X29选自Leu、Ile、Val或Phe。
2.根据权利要求1所述的甲状旁腺激素相关肽类似物,其特征在于,所提供的PTHrP类似物选自氨基酸序列如SEQ ID NO.4所示的化合物1、氨基酸序列如SEQ ID NO.5所示的化合物2、氨基酸序列如SEQ ID NO.6所示的化合物3或氨基酸序列如SEQ ID NO.7所示的化合物4。
3.权利要求1或2所述的甲状旁腺激素相关肽类似物在制备用于预防和/或治疗骨骼疾病及其相关并发症药物中的应用。
4.根据权利要求3所述的应用,其特征在于,所述的预防和/或治疗骨骼疾病及其相关并发症药物包括预防和/或治疗骨质疏松症的药物组合物。
5.一种用于预防和/或治疗骨质疏松症的药物组合物,其特征在于,由有效量化合物和药学上可接受的辅料组成;
所述的有效量化合物为权利要求1或2所述的甲状旁腺激素相关肽类似物,或是权利要求1或2所述的甲状旁腺激素相关肽类似物与其他治疗骨质疏松疾病的药物的组合。
6.根据权利要求5所述的用于预防和/或治疗骨质疏松症的药物组合物,其特征在于,
所述的辅料包括稀释剂、赋形剂、润滑剂、粘合剂、填充剂、防腐剂、表面活性剂、着色剂、矫味剂、乳化剂、助悬剂、胶凝剂、崩解剂、pH调节剂和增溶剂中的至少一种。
7.根据权利要求6所述的用于预防和/或治疗骨质疏松症的药物组合物,其特征在于,
所述的辅料用于辅助将所述的药物组合物制成不同的剂型,包括但不限于:注射剂或冻干粉、片剂、贴剂、丸剂、锭剂、软胶囊剂、硬胶囊剂、颗粒剂、散剂、溶液剂、混悬剂和糖浆剂。
8.根据权利要求5所述的用于预防和/或治疗骨质疏松症的药物组合物,其特征在于,所述的其他治疗骨质疏松疾病的药物包括双磷酸盐类、降钙素、***类、选择性***受体调节剂类、锶盐类、活性维生素D及其类似物和护骨强骨的中药类药物中的至少一种。
9.根据权利要求5~8任一项所述的用于预防和/或治疗骨质疏松症的药物组合物,其特征在于,所述的药物组合物的给药方式包括口服给药、吸入给药和肠胃外给药中的至少一种或几种;
所述的药物组合物的给药频率为至少一天给药一次、一周给药一次或一个月给药一次。
10.根据权利要求9所述的用于预防和/或治疗骨质疏松症的药物组合物,其特征在于,所述的肠胃外给药通过腹膜内、肌肉内、动脉内、静脉内、皮下、皮内注射或透皮微针方式给药。
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5589452A (en) * | 1992-07-14 | 1996-12-31 | Syntex (U.S.A.) Inc. | Analogs of parathyroid hormone and parathyroid hormone related peptide: synthesis and use for the treatment of osteoporosis |
CN1193915A (zh) * | 1995-08-29 | 1998-09-23 | 森德克斯(美国)股份有限公司 | 用于治疗骨质疏松症化合物的鼻传递的药物组合物 |
-
2020
- 2020-08-06 CN CN202010780705.5A patent/CN114057863B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5589452A (en) * | 1992-07-14 | 1996-12-31 | Syntex (U.S.A.) Inc. | Analogs of parathyroid hormone and parathyroid hormone related peptide: synthesis and use for the treatment of osteoporosis |
CN1193915A (zh) * | 1995-08-29 | 1998-09-23 | 森德克斯(美国)股份有限公司 | 用于治疗骨质疏松症化合物的鼻传递的药物组合物 |
Non-Patent Citations (2)
Title |
---|
LIU S, JEAN-ALPHONSE FG, WHITE AD, WOOTTEN D, SEXTON PM, GARDELLA TJ, VILARDAGA JP, GELLMAN SH.: "Use of Backbone Modification To Enlarge the Spatiotemporal Diversity of Parathyroid Hormone Receptor-1 Signaling via Biased Agonism", 《J AM CHEM SOC.》, pages 14486 - 14490 * |
施小明;王春晓;庄志华;吴洁;李泰明;曹荣月;刘景晶;: "一种新型hPTH(1-34)基因相关肽的制备及其应用的初步研究", 东南大学学报(医学版), no. 01, pages 31 - 36 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2024032457A1 (zh) * | 2022-08-10 | 2024-02-15 | 成都奥达生物科技有限公司 | 一种长效特立帕肽化合物 |
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