CN108026565A - 用于鉴定ido1和/或tdo调节剂的筛选测定法 - Google Patents
用于鉴定ido1和/或tdo调节剂的筛选测定法 Download PDFInfo
- Publication number
- CN108026565A CN108026565A CN201680053185.XA CN201680053185A CN108026565A CN 108026565 A CN108026565 A CN 108026565A CN 201680053185 A CN201680053185 A CN 201680053185A CN 108026565 A CN108026565 A CN 108026565A
- Authority
- CN
- China
- Prior art keywords
- tdo
- ido1
- cell
- method based
- cell described
- 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
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
- G01N33/5041—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects involving analysis of members of signalling pathways
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
- G01N2333/90241—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明提供了用于鉴定吲哚胺2,3‑双加氧酶1(IDO1)和/或色氨酸2,3‑双加氧酶(TDO)调节剂的基于细胞的方法。
Description
技术领域
本发明涉及用于鉴定吲哚胺2,3-双加氧酶1(IDO1)和/或色氨酸2,3-双加氧酶(TDO)特异性调节剂的基于细胞的筛选测定法。
背景技术
吲哚胺2,3-双加氧酶1(IDO1)和色氨酸2,3-双加氧酶(TDO)是细胞质的含血红素的酶,其催化色氨酸(Trp)氧化裂解成N-甲酰犬尿氨酸(NFK),这是犬尿氨酸(Kyn)途径的第一步。Trp的消耗和Kyn代谢物的形成导致效应T细胞功能的抑制和调节性T细胞的分化。肿瘤细胞中IDO1和/或TDO表达的增加水平与癌症中存活预后不良相关。IDO已被临床验证为癌症的小分子药物靶标,而临床前研究表明TDO可能是癌症免疫疗法的靶标(参见etal,2009;Platten et al,2014的综述及其中的参考文献)。
Pilotte等,2012)描述了用于测试IDO1和TDO抑制剂的低通量质谱和高效液相色谱(HPLC)测定法。
本发明要解决的问题是提供用于鉴定IDO/TDO特异性调节剂的高通量兼容测定法。
概述
本发明提供了用于鉴定吲哚胺2,3-双加氧酶1(IDO1)和/或色氨酸2,3-双加氧酶(TDO)调节剂的基于细胞的方法,包括:
a.提供重组表达IDO1和/或TDO的细胞,其中所述IDO1和/或TDO表达是可诱导的,
b.使步骤a)的细胞与测试化合物和IDO1和/或TDO底物接触,
c.使步骤b)的混合物与犬尿氨酸感受器接触,
d.测量步骤c)的混合物的荧光读数,其中在候选化合物存在下与空白相比改变的荧光读数指示IDO1和/或TDO活性的调节剂。
在本发明的一个具体实施方案中,细胞是HepG2细胞。
在本发明的一个具体实施方案中,诱导型IDO1和/或TDO表达是由于Tet-on系统。
在本发明的一个具体实施方案中,犬尿氨酸感受器是7-(二乙基氨基)-4-乙基硫基-2-氧代-色烯-3-甲醛(感受器1)。
在本发明的一个具体实施方案中,荧光在Ex:520-560nm和Em:580-680nm测量。
在本发明的一个具体实施方案中,未诱导的细胞被用作空白。
在本发明的具体实施方案中,IDO1和/或TDO是人IDO1和人TDO。
在本发明的一个具体实施方案中,重组表达IDO1和/或TDO的细胞是稳定的细胞系。
在本发明的具体实施方案中,在步骤d)中测量步骤c)的混合物的上清液的荧光读数。
在本发明的一个具体实施方案中,该方法在微量滴定板中进行。
在本发明的一个具体实施方案中,与空白相比,候选化合物存在下的荧光读数降低指示IDO1和/或TDO抑制剂。
在本发明的一个具体实施方案中,与空白相比,候选化合物存在下荧光读数增加表明IDO1和/或TDO激活剂。
在本发明的一个具体实施方案中,IDO1和/或TDO底物是色氨酸。
所描述的测定法旨在筛选/描绘和发现新的,高度有效的IDO/TDO选择性和/或双重调节剂,其是可以帮助破坏肿瘤微环境内的免疫耐受性并且防止肿瘤逃脱免疫监督和破坏的免疫治疗剂。
附图简述
图1显示了本发明的基于IDO1/TDO细胞的筛选测定法的图示。
图2A显示HepG2-Tet-on IDO1*29细胞中IDO1的剂量依赖性诱导。用小鼠抗IDO1mAb(*UM500091,Origene)免疫印迹来自HepG2-Tet-on-IDO1*29的总细胞裂解物。用指定浓度的多西环素处理细胞24小时。
图2B显示了HepG2-Tet-on IDO1*25细胞中TDO的剂量依赖性诱导。用小鼠抗TDO2mAb(*TA504730,Origene)免疫印迹来自HepG2-Tet-on-TDO*25的总细胞裂解物。用指定浓度的多西环素处理细胞24小时。
图3A:将HepG2-Tet-on-IDO1*29细胞用指定浓度的多西环素处理24小时。如测定方法中所述进行INCB024360的测定。INCB024360=(Z)-N-(3-溴-4-氟苯基)-N'-羟基-4-(2-(氨磺酰基氨基)乙基氨基)-1,2,5-二唑-3-甲脒。WO2010005958中的实施例1。
图3B.用指定浓度的多西环素处理HepG2-Tet-on-TDO*25细胞24小时。如测定方法中所述进行INCB024360的测定。
本发明实施方案详述
建立了一种新型的基于细胞的荧光测定法来测量IDO1/TDO活性,该活性适合用于IDO和/或TDO调节剂的化合物文库的高通量筛选。该测定法开辟了化学空间的新领域,用于发现两种重要药物靶点的抑制剂。它依靠通过荧光化学感受器(感受器1)(Klockow andGlass,2013)定量测定介质中产生的犬尿氨酸的量,并补充标准低通量质谱和高效液相色谱(HPLC)测定方法(Pilotte等,2012)。
术语“IDO1”在本文中用于指来自任何动物(例如,包括人在内的哺乳动物物种)的吲哚胺2,3-双加氧酶1的天然序列和IDO1变体(其在下文进一步定义)。
“天然序列IDO1”是指与天然存在的IDO1多肽具有相同氨基酸序列的多肽,而不管其制备方式如何。天然序列IDO1可以从自然界分离,或通过重组和/或合成方法制备。术语“天然序列IDO1”具体包括天然存在的截短或分泌形式,天然存在的变体形式(例如选择性剪接形式)和IDO1的天然存在的等位基因变体。人IDO1多肽的氨基酸序列在Seq.Id.No.1给出。
术语“IDO1变体”是指天然序列IDO1的氨基酸序列变体,其在天然序列中含有一个或多个氨基酸取代和/或缺失和/或插入。氨基酸序列变体通常与天然序列IDO1的氨基酸序列具有至少约75%,优选至少约80%,更优选至少约85%,甚至更优选至少约90%,最优选至少约95%的序列同一性。
术语“TDO”在本文中用于指来自任何动物(例如,包括人在内的哺乳动物物种)的吲哚胺2,3-双加氧酶的天然序列,和TDO变体(其在下文进一步定义)。
“天然序列TDO”是指与天然存在的TDO多肽具有相同氨基酸序列的多肽,而不管其制备方式如何。天然序列TDO可以从自然界分离,或通过重组和/或合成方法制备。术语“天然序列TDO”具体包括天然存在的截短或分泌形式,天然存在的变体形式(例如选择性剪接形式)和TDO的天然存在的等位基因变体。人类TDO多肽的氨基酸序列在Seq.Id.No.2给出。
术语“TDO变体”是指天然序列TDO的氨基酸序列变体,其在天然序列含有一个或多个氨基酸取代和/或缺失和/或插入。氨基酸序列变体通常与天然序列TDO的氨基酸序列具有至少约75%,优选至少约80%,更优选至少约85%,甚至更优选至少约90%,最优选至少约95%的序列同一性。
术语“化合物”在本文中用于结合本发明的测定描述的“测试化合物”或“药物候选化合物”的上下文中。如此,这些化合物包含合成衍生的或天然来源的有机或无机化合物。所述化合物包括无机或有机化合物,例如特征在于相对低分子量的多核苷酸,脂质或激素类似物。其他生物聚合物有机测试化合物包括包含约2至约40个氨基酸的肽和包含约40至约500个氨基酸的更大多肽,如抗体或抗体缀合物。
实施例
犬尿氨酸感受器7-(二乙基氨基)-4-乙基硫基-2-氧代-色烯-3-甲醛的合成(感受器1)
已经描述了用于检测犬尿氨酸的感受器1的合成和开发(Klockow and Glass,2013)。
犬尿氨酸的感受器1的合成和开发
产生HepG2稳定细胞系,允许以多西环素依赖性方式诱导性表达人IDO1或人TDO2
将HepG2细胞在MEM(1X)+GlutaMax(*41090,Building 92-储备液),10%FBS(*16000-044,)中培养。使用质粒pTet-On(*631018,Clontech)和Lipofectamine 2000(*11668019,Life Technology)进行稳定转染的第一步,以建立表达反向四环素依赖性反式激活蛋白的细胞。用10μg pTet-On转染10cm培养皿中的HepG2细胞,然后用400μg/ml G418选择21天,用克隆环分离抗性菌落。通过瞬时转染具有由Tet-On启动子驱动的萤光素酶基因的报告质粒pTRE2-Luc(*S1496,Clontech)来测试各个克隆的反向四环素依赖性反式激活蛋白的表达。选择表现出非常高的四环素诱导型萤光素酶活性和不可检测的基础萤光素酶活性的一个克隆系并用于用IDO1或TDO2表达质粒进行第二轮转染。通过将人IDO1或TDO2cDNA(均购自Origene)插入pTRE2hyg表达载体(*631014,Clontech)中来构建这些质粒。在用400μg/ml潮霉素进行二次稳定转染和选择后,将个体抗性菌落克隆并保持用400μg/ml G418和400μg/ml潮霉素的长期培养。通过蛋白质印迹筛选克隆并且基于在与1μg/ml多西环素在不可检测但非诱导条件下细胞培养24小时后对IDO或TDO蛋白的高诱导选择两个命名为HepG2-Tet-on-IDO1*29和HepG2-Tet-on-TDO*25的稳定克隆。
INCB024360化合物=(Z)-N-(3-溴-4-氟苯基)-N'-羟基-4-(2-(氨磺酰基氨基)乙基氨基)-1,2,5-二唑-3-甲脒。INCB024630是WO2010005958中的实施例1。
测定方法:
将细胞以10,000细胞/孔接种在细胞板中,并在95%湿润的细胞培养箱中在37℃和5%CO2下温育。然后用1μg/ml多西环素诱导细胞24小时以实现IDO或TDO的完全表达。未诱导的细胞用作100%抑制对照。用60μl 1X HBSS(37℃)洗涤3次后,用细胞培养箱中的60μl 1X HBSS平衡细胞30分钟。再用60μl1X HBSS(37℃)洗涤3次后,将细胞用化合物处理10分钟,然后加入底物L-色氨酸至最终浓度为80μM。该测定在细胞培养箱中在37℃和5%CO2下进行4小时。通过将30μl/孔的上清液转移到测定板中来终止反应。加入10μl/孔的30%(w/v)TCA,接着施加感受器1至最终浓度为10μM。短暂离心后,用Paradigm(Moleculardevices)读板仪在Ex:546,Em:586nm处测量荧光。
测定材料:
平板
测定板Costar 384孔,全部澄清,NT,*3702
细胞板Costar 384孔,全部澄清,PDL包被的,*356662细胞和缓冲液
本发明涉及使用允许IDO或TDO的诱导型表达的内部建立的HepG2稳定细胞系的新型HTS-兼容细胞测定法。HepG2细胞不表达内源性IDO1和TDO(Pilotte等,2012),因此这些酶可以以严格控制的多西环素-剂量依赖性方式诱导,如蛋白质印迹所证明的(图2A和2B)。诱导的IDO1或TDO将色氨酸转化为N-甲酰基犬尿氨酸,然后通过丰富的甲酰胺酶将其代谢成犬尿氨酸。大部分产生的犬尿氨酸被释放到培养基中,通过荧光“感受器I”探针分析所述培养基(图1)。现有技术的生物化学IDO1,但不是现有技术的生物化学TDO测定,是从酶测定法翻译成基于细胞的测定法,证实了文献报道(Liu等人,2010)。在使用参考化合物INCB024360的TDO酶活性中观察到从生物化学测定到基于细胞的测定的IC50右移100多倍。因此,基于细胞的测定法对于化合物IC50的测定更具生物学相关性和可靠性。
本发明的测定法不仅是用于IDO或TDO的新型HTS-兼容的基于细胞的测定法,而且与文献(Liu等人,2010)相比还产生了更可靠的测定窗口。如图3A和3B所示,测定窗口虽然不是IC50,但依赖于细胞中IDO和TDO的表达水平。众所周知,与大多数哺乳动物细胞中内源或病毒启动子介导的表达相比,Tet-on可诱导系统允许更高的转基因表达(Gossen等,1995)。
参考文献
Gossen M.,Freundlieb S.,Bender G.,Muller G.,Hillen W.,Bujard H.(1995)Transcriptional activation by tetracyclines in mammalian cells,Science 268:1766–1769.
Hwu P.,Du M.X.,Lapointe R.,Do M.,Taylor M.W.,Young H.A.(2000),Indoleamine 2,3-dioxygenase production by human dendritic cells results inthe inhibition of T cell proliferation.The Journal of Immunology,164:3596-3599
Klockow J.L.and Glass T.E.(2013),Development of a FluorescentChemosensor for the Detection of Kynurenine,Organic Letters,15(2):235–237
S., A.,Rammensee H-G.,Opelz G.and Terness P.(2009),Inhibitors of indoleamine-2,3-dioxygenase for cancer therapy:can wesee the wood for the trees?,Nature Reviews/Cancer,9:445-452.
Liu X.,Shin N.,Koblish H.K.,Yang G.,Wang Q.,Wang K.,Leffet L.,Hansbury M.J.,Thomas B.,Rupar M.,Waeltz P.,Bowman K.J.,Polam P.,Sparks R.B.,Yue E.W.,Li Y.,Wynn R.,Fridman J.S.,Burn T.C.,Combs A.P.,Newton R.C.,ScherleP.A.(2010),Selective inhibition of IDO1 effectively regulates mediators ofantitumor immunity,Blood,115(17):3520-30.
Pilotte L.,Larrieua P.,Stroobanta V.,Colaua D., E.,Frédérickb R.,De Plaena E.,Uyttenhovea C.,Woutersb J.,Masereelb B.,and Van denEyndea B.J.(2012)Reversal of tumoral immune resistance by inhibition oftryptophan 2,3-dioxygenase,PNAS,109(7):2497–2502
Platten M.,von Knebel Doeberitz N.,Oezen I.,Wick W.and Ochs K.(2014),Cancer immunotherapy by targeting IDO1/TDO and their downstream effectors,Frontiers in Immunology,5(673):1-7
Claims (14)
1.用于鉴定吲哚胺2,3-双加氧酶1(IDO1)和/或色氨酸2,3-双加氧酶(TDO)调节剂的基于细胞的方法,包括:
a.提供重组表达IDO1和/或TDO的细胞,其中所述IDO1和/或TDO表达是可诱导的,
b.使步骤a)的细胞与测试化合物和IDO1和/或TDO底物接触,
c.使步骤b)的混合物与犬尿氨酸感受器接触,和
d.测量步骤c)的混合物的荧光读数,其中在候选化合物存在下与空白相比改变的荧光读数指示IDO1和/或TDO活性的调节剂。
2.权利要求1所述的基于细胞的方法,其中所述细胞是HepG2细胞。
3.权利要求1或2所述的基于细胞的方法,其中可诱导的IDO1和/或TDO表达是由于Tet-on系统。
4.权利要求1至3所述的基于细胞的方法,其中所述犬尿氨酸感受器是7-(二乙基氨基)-4-乙基硫基-2-氧代-色烯-3-甲醛(感受器1)。
5.权利要求4所述的基于细胞的方法,其中所述荧光在激发:520-560nm和发射:580-680nm下测量。
6.权利要求1至5所述的基于细胞的方法,其中未诱导的细胞被用作空白。
7.权利要求1至6所述的基于细胞的方法,其中所述IDO1和/或TDO是人IDO1和人TDO。
8.权利要求1至7的基于细胞的方法,其中重组表达IDO1和/或TDO的细胞是稳定的细胞系。
9.权利要求1至8所述的基于细胞的方法,其中在步骤d)中,测量步骤c)的混合物的上清液的荧光读数。
10.权利要求1至9所述的基于细胞的方法,其中所述方法在costar 384孔板中进行。
11.权利要求1至10所述的基于细胞的方法,其中与空白相比在候选化合物存在下降低的荧光读数指示IDO1和/或TDO抑制剂。
12.权利要求1至10所述的基于细胞的方法,其中与空白相比在候选化合物存在下增加的荧光读数指示IDO1和/或TDO激活剂。
13.权利要求1至12所述的基于细胞的方法,其中所述IDO1和/或TDO底物是色氨酸。
14.权利要求1至13所述的基于细胞的方法,其中所述方法是自动化的。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP15193667.1 | 2015-11-09 | ||
EP15193667 | 2015-11-09 | ||
PCT/EP2016/076765 WO2017080934A1 (en) | 2015-11-09 | 2016-11-07 | Screening assay to identify id01 and/or tdo modulators |
Publications (1)
Publication Number | Publication Date |
---|---|
CN108026565A true CN108026565A (zh) | 2018-05-11 |
Family
ID=54539884
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201680053185.XA Pending CN108026565A (zh) | 2015-11-09 | 2016-11-07 | 用于鉴定ido1和/或tdo调节剂的筛选测定法 |
Country Status (6)
Country | Link |
---|---|
US (1) | US20180252704A1 (zh) |
EP (1) | EP3374517A1 (zh) |
JP (1) | JP2018532406A (zh) |
CN (1) | CN108026565A (zh) |
HK (1) | HK1254241A1 (zh) |
WO (1) | WO2017080934A1 (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109706180A (zh) * | 2019-01-11 | 2019-05-03 | 杭州荣泽生物科技有限公司 | 一种脐带间充质干细胞过表达ido增强免疫抑制的方法及应用 |
CN110317855A (zh) * | 2019-07-26 | 2019-10-11 | 中国药科大学 | 基于细胞的鉴定ido1酶活性及筛选ido1酶抑制剂的荧光检测方法 |
CN113278674A (zh) * | 2021-07-08 | 2021-08-20 | 华夏源(上海)生命科技有限公司 | 一种快速检测人间充质干细胞ido1活性的实验方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004093871A1 (en) * | 2003-03-27 | 2004-11-04 | Lankenau Institute For Medical Research | Novel methods for the treatment of cancer |
WO2008068621A2 (en) * | 2006-12-05 | 2008-06-12 | Molmed Spa | Combination product with ido inhibitor and tumor targeted ifn-gamma |
WO2008143668A2 (en) * | 2006-05-18 | 2008-11-27 | Lankenau Institute For Medical Research | Indoleamine-2, 3-dioxygenase-2 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101927291B1 (ko) | 2008-07-08 | 2018-12-10 | 인사이트 홀딩스 코포레이션 | 인돌아민 2,3-디옥시게나아제의 억제제로서의 1,2,5-옥사디아졸 |
NO2694640T3 (zh) * | 2011-04-15 | 2018-03-17 |
-
2016
- 2016-11-07 CN CN201680053185.XA patent/CN108026565A/zh active Pending
- 2016-11-07 EP EP16794267.1A patent/EP3374517A1/en not_active Withdrawn
- 2016-11-07 JP JP2018519869A patent/JP2018532406A/ja active Pending
- 2016-11-07 WO PCT/EP2016/076765 patent/WO2017080934A1/en active Application Filing
-
2018
- 2018-05-08 US US15/973,837 patent/US20180252704A1/en not_active Abandoned
- 2018-10-18 HK HK18113384.7A patent/HK1254241A1/zh unknown
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004093871A1 (en) * | 2003-03-27 | 2004-11-04 | Lankenau Institute For Medical Research | Novel methods for the treatment of cancer |
WO2008143668A2 (en) * | 2006-05-18 | 2008-11-27 | Lankenau Institute For Medical Research | Indoleamine-2, 3-dioxygenase-2 |
WO2008068621A2 (en) * | 2006-12-05 | 2008-06-12 | Molmed Spa | Combination product with ido inhibitor and tumor targeted ifn-gamma |
Non-Patent Citations (3)
Title |
---|
JESSICA L.KLOCKOW等: "Development of a Fluorescent Chemosensor for the Detection of Kynurenine", 《ORGANIC LETTERS》 * |
NONE: "Amabio:hIDO1-HEK293 Recombinant Cell", 《HTTP://WWW.AMSBIO.COM/DATASHEETS/60532.PDF.》 * |
UTE F. RÖHRIG等: "Challenges in the Discovery of Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitors", 《J. MED. CHEM.》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109706180A (zh) * | 2019-01-11 | 2019-05-03 | 杭州荣泽生物科技有限公司 | 一种脐带间充质干细胞过表达ido增强免疫抑制的方法及应用 |
CN110317855A (zh) * | 2019-07-26 | 2019-10-11 | 中国药科大学 | 基于细胞的鉴定ido1酶活性及筛选ido1酶抑制剂的荧光检测方法 |
CN113278674A (zh) * | 2021-07-08 | 2021-08-20 | 华夏源(上海)生命科技有限公司 | 一种快速检测人间充质干细胞ido1活性的实验方法 |
Also Published As
Publication number | Publication date |
---|---|
HK1254241A1 (zh) | 2019-07-12 |
US20180252704A1 (en) | 2018-09-06 |
WO2017080934A1 (en) | 2017-05-18 |
JP2018532406A (ja) | 2018-11-08 |
EP3374517A1 (en) | 2018-09-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Bugaj et al. | Optogenetic protein clustering and signaling activation in mammalian cells | |
Moroz-Omori et al. | Photoswitchable gRNAs for spatiotemporally controlled CRISPR-Cas-based genomic regulation | |
Mackintosh et al. | The molecular pathogenesis of Ewing sarcoma | |
WO2020036973A1 (en) | Methods and compositions for treating mitochondrial disease or disorders and heteroplasmy | |
CN108601849A (zh) | 一种在淋巴细胞中高水平的和稳定的基因转移的方法 | |
Jin et al. | RNA‐binding motif protein 24 regulates myogenin expression and promotes myogenic differentiation | |
Von Werder et al. | Production of avian retroviruses and tissue-specific somatic retroviral gene transfer in vivo using the RCAS/TVA system | |
US20170292961A1 (en) | Systems and methods for assessing inter-cell communication | |
AU2003239947A1 (en) | Heterologous stimulus-gated ion channels and methods of using same | |
CN108026565A (zh) | 用于鉴定ido1和/或tdo调节剂的筛选测定法 | |
Den Hartogh et al. | Concise review: fluorescent reporters in human pluripotent stem cells: contributions to cardiac differentiation and their applications in cardiac disease and toxicity | |
Körber et al. | Effects of distinct collybistin isoforms on the formation of GABAergic synapses in hippocampal neurons | |
Lamparter et al. | Phytochromes from Agrobacterium fabrum | |
Leschik et al. | Embryonic stem cells stably expressing BDNF–GFP exhibit a BDNF-release-dependent enhancement of neuronal differentiation | |
Sun et al. | Deep single-cell-type proteome profiling of mouse brain by nonsurgical AAV-mediated proximity labeling | |
Mondal et al. | Repurposing protein degradation for optogenetic modulation of protein activities | |
Belletti et al. | Regulation of Id2 gene expression by the insulin-like growth factor I receptor requires signaling by phosphatidylinositol 3-kinase | |
Li et al. | Efficient photoactivatable Dre recombinase for cell type-specific spatiotemporal control of genome engineering in the mouse | |
Zeng et al. | Targeted addition of mini-dystrophin into rDNA locus of Duchenne muscular dystrophy patient-derived iPSCs | |
Zheng et al. | Modification of Tet1 and histone methylation dynamics in dairy goat male germline stem cells | |
Nazlamova et al. | Generation of a cone photoreceptor-specific GNGT2 reporter line in human pluripotent stem cells | |
Poggi | Generation of tumor spheroids to evaluate T cell and NK cell cytotoxicity | |
KR101620812B1 (ko) | CLC-Kb 염화 이온 채널 조절물질 도출을 위한 세포기반 형광 이미징 고효율 검색 및 광범위 특성 연구용 세포주 | |
Chan et al. | Cell type-and stage-specific expression of Otx2 is coordinated by a cohort of transcription factors and multiple cis-regulatory modules in the retina | |
Franck et al. | Mechanosensitive clathrin platforms anchor desmin intermediate filaments in skeletal muscle |
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 | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1254241 Country of ref document: HK |
|
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180511 |
|
WD01 | Invention patent application deemed withdrawn after publication |