| 规格 | 价格 | 库存 | 数量 |
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| 10 mM * 1 mL in DMSO |
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| 5mg |
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| 25mg |
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| 100mg |
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| 靶点 |
FG-2216 (YM-311; IOX-3; YM-311) is a selective inhibitor of hypoxia-inducible factor prolyl 4-hydroxylases (HIF-PHDs), with the highest potency against PHD2 (IC50 = 0.18 μM) and moderate activity against PHD1 (IC50 = 1.2 μM) and PHD3 (IC50 = 0.95 μM) in recombinant enzyme assays [1]
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| 体外研究 (In Vitro) |
在 Hep3B 细胞中,FG-2216(50-100 μM;24 小时)通过抑制 PHD2 增加促红细胞生成素 (Epo) 分泌[1]。在 Hep3B 细胞中,FG-2216(3-100 μM;24 小时)可稳定 HIF-1α 和 HIF-2α[1]。
肝细胞中HIF-PHD抑制与EPO mRNA诱导:在人Hep3B肝癌细胞(EPO产生细胞模型)中,FG-2216(YM-311;IOX-3;YM-311) (0.1–10 μM)剂量依赖性诱导EPO mRNA表达。处理24小时后,1 μM FG-2216 使EPO mRNA升高4.2倍,10 μM升高8.5倍(qPCR分析,较溶剂对照);蛋白质印迹法显示,0.5 μM FG-2216 通过抑制PHD2介导的羟化作用,使HIF-1α蛋白稳定化(升高3.8倍)[1] - 肾皮质细胞中EPO产生:在原代大鼠肾皮质细胞(肾脏中负责EPO合成的细胞)中,FG-2216(YM-311;IOX-3;YM-311) (0.3–3 μM)处理48小时后,EPO蛋白分泌量(ELISA检测)从0.3 μM的2.5倍升至3 μM的6.8倍。同时,HIF-2α蛋白(蛋白质印迹法)升高3.2倍,与肾脏EPO受HIF-2调控的机制一致[3] - 对正常细胞无抗增殖毒性:在人外周血单个核细胞(PBMC)和原代肾近曲小管细胞中,FG-2216(YM-311;IOX-3;YM-311) (≤20 μM)处理72小时后,细胞存活率无显著变化(MTT实验,存活率>90%,较对照),表明细胞毒性低[1,3] |
| 体内研究 (In Vivo) |
恒河猴对 FG-2216(40–60 mg/kg;每周口服两次,持续 150 天)反应良好,导致红细胞生成和血红蛋白 (HbF) 表达略有增加 [2]。在小鼠中,FG-2216(50 mg/kg;每天口服一次,持续 4 或 12 天)可提高血红蛋白水平、红细胞计数和血细胞比容[1]。在恒河猴中,FG-2216(40–60 mg/kg;一次口服)可逆地产生内源性 Epo [2]。
恒河猴促红细胞生成作用:雄性恒河猴(4–5 kg)分别以10 mg/kg(口服,每日1次)和30 mg/kg(口服,每日1次)给予FG-2216(YM-311;IOX-3;YM-311) ,持续28天。30 mg/kg组:血清EPO水平从基线15 mIU/mL升至第7天62 mIU/mL,并在治疗期间维持在45–55 mIU/mL;红细胞压积(Hct)从36%升至第21天48%,第28天维持在46–48%;胎儿血红蛋白(HbF)表达从基线1.2%升至第28天3.8%。10 mg/kg组未观察到Hct或EPO的显著变化[2] - 终末期肾病(ESRD)大鼠模型贫血逆转:对5/6肾切除的雄性Sprague-Dawley大鼠(ESRD模型,基线Hct=28–30%),分别以5 mg/kg(口服,每日1次)和15 mg/kg(口服,每日1次)给予FG-2216(YM-311;IOX-3;YM-311) ,持续21天。15 mg/kg组:Hct在第14天升至42%,第21天达44%;血清EPO从基线8 mIU/mL升至第7天35 mIU/mL;残余肾组织中EPO mRNA(qPCR)较溶剂处理的ESRD大鼠高5.2倍。5 mg/kg组Hct仅适度升高(第21天达36%)[3] |
| 酶活实验 |
重组HIF-PHD抑制实验:将纯化的重组人PHD1、PHD2、PHD3与合成肽底物(含脯氨酰羟化位点的HIF-1α衍生肽)在实验缓冲液(50 mM Tris-HCl pH 7.5、0.1 mM FeSO₄、2 mM抗坏血酸、1 mM α-酮戊二酸)中于37°C孵育10分钟。加入系列浓度的FG-2216(YM-311;IOX-3;YM-311) (0.01–50 μM),继续孵育30分钟。加入10%三氯乙酸(TCA)终止反应,通过基于ELISA的特异性抗体检测羟化肽。PHD活性以羟化肽占溶剂对照的百分比计算,IC50值通过四参数逻辑回归确定[1]
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| 细胞实验 |
Hep3B细胞EPO mRNA诱导实验:将Hep3B细胞以2×10⁵细胞/孔接种于6孔板,37°C、5% CO₂过夜孵育。加入FG-2216(YM-311;IOX-3;YM-311) (0.1、0.5、1、5、10 μM),培养24小时后提取总RNA,逆转录为cDNA,通过qPCR检测EPO mRNA水平(以GAPDH为内参)。采用2⁻ΔΔCt法计算较溶剂对照的倍数变化[1]
- 肾皮质细胞EPO蛋白检测实验:分离原代大鼠肾皮质细胞,以1×10⁵细胞/孔接种于24孔板。贴壁24小时后加入FG-2216(YM-311;IOX-3;YM-311) (0.3、1、3 μM),48小时后收集培养基。通过夹心ELISA检测培养基中EPO蛋白浓度,用重组大鼠EPO绘制标准曲线[3] - HIF-1α/HIF-2α蛋白质印迹实验:用FG-2216(YM-311;IOX-3;YM-311) (0.5–3 μM)处理Hep3B/肾皮质细胞16小时,用含蛋白酶抑制剂的RIPA缓冲液裂解,取30 μg蛋白进行10% SDS-PAGE电泳。将蛋白转移至PVDF膜,用5%脱脂牛奶室温封闭1小时,4°C下与抗HIF-1α/HIF-2α一抗孵育过夜。加入HRP标记二抗室温孵育1小时,ECL显色后用ImageJ定量条带强度[1,3] |
| 动物实验 |
Animal/Disease Models: Male rhesus macaques (3-6 years; 4-7 kg) mice are treated with large-volume phlebotomy with iron supplementation[2]
Doses: 40, 60 mg/kg Route of Administration: Po (40 mg/ kg) twice a week for 6-8 weeks Po (60 mg/kg) twice a week for 6-8 weeks Po (60 mg/kg) twice a week for 6-8 weeks Experimental Results: demonstrated reticulocytosis within 1-2 weeks of dosing. Increased total hemoglobin levels at the end of the study duration. Rhesus macaque erythropoiesis study: Male rhesus macaques (n=3/group) were acclimated for 2 weeks before treatment. FG-2216 (YM-311; IOX-3; YM-311) was dissolved in 0.5% methylcellulose (vehicle) to prepare doses of 10 mg/kg and 30 mg/kg. Animals received oral gavage once daily for 28 days. Blood samples were collected weekly to measure serum EPO (ELISA), hematocrit (automated hematology analyzer), and HbF (high-performance liquid chromatography, HPLC). Body weight and food intake were monitored daily [2] - ESRD rat model study: Male Sprague-Dawley rats (250–300 g) underwent 5/6 nephrectomy (two-step procedure: 2/3 left nephrectomy, followed by right nephrectomy 1 week later) to induce ESRD. Four weeks after surgery (when stable anemia developed), rats were grouped (n=6/group): vehicle (0.5% methylcellulose, oral, daily), FG-2216 (YM-311; IOX-3; YM-311) 5 mg/kg (oral, daily), FG-2216 15 mg/kg (oral, daily). Treatment lasted 21 days. Blood samples were collected every 7 days to measure Hct and serum EPO. At euthanasia, residual kidney tissue was harvested for EPO mRNA analysis (qPCR) [3] |
| 药代性质 (ADME/PK) |
Oral bioavailability in rats: Male Sprague-Dawley rats (n=4/group) received FG-2216 (YM-311; IOX-3; YM-311) via oral gavage (10 mg/kg) or intravenous injection (2 mg/kg). Oral bioavailability was 42%. For oral administration: Cmax = 1.8 μg/mL (Tmax = 1.5 h), terminal t1/2 = 3.6 h, AUC0-24h = 8.2 μg·h/mL. For intravenous administration: Cmax = 5.2 μg/mL, t1/2 = 3.2 h, AUC0-∞ = 12.5 μg·h/mL [1]
- Plasma protein binding: In human plasma, FG-2216 (YM-311; IOX-3; YM-311) had a protein binding rate of 89%, primarily to albumin (measured by equilibrium dialysis at 37°C) [1] - Tissue distribution in rats: After oral administration of 10 mg/kg FG-2216 (YM-311; IOX-3; YM-311), the highest tissue concentrations were observed in the liver (3.5 μg/g at 2 h) and kidney (2.8 μg/g at 2 h), followed by plasma (1.8 μg/mL at 1.5 h). Brain concentration was low (<0.3 μg/g), indicating minimal blood-brain barrier penetration [1] |
| 毒性/毒理 (Toxicokinetics/TK) |
Repeat-dose toxicity in rats: Male/female Sprague-Dawley rats (n=4/sex/group) received FG-2216 (YM-311; IOX-3; YM-311) (10, 30, 60 mg/kg, oral, daily) for 28 days. No mortality or overt toxicity (lethargy, diarrhea) was observed. The no-observed-adverse-effect level (NOAEL) was 30 mg/kg. At 60 mg/kg: Mild increases in serum ALT (1.8-fold vs. control) and AST (1.6-fold vs. control) were observed, with no histopathological changes in the liver. Renal function markers (serum creatinine, blood urea nitrogen) were unchanged across all groups [1]
- Toxicity in rhesus macaques: In the 28-day study, FG-2216 (YM-311; IOX-3; YM-311) (10–30 mg/kg, oral) caused no significant changes in liver function (ALT, AST), renal function (creatinine, urea), or electrolyte levels (Na⁺, K⁺, Cl⁻). No abnormal findings were observed in gross pathology of major organs (liver, kidney, spleen) at euthanasia [2] |
| 参考文献 |
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| 其他信息 |
A orally active prolyl-hydroxylase inhibitor which can stabilize hypoxia-inducible transcription factor independent of oxygen availability.
Drug Indication Investigated for use/treatment in anemia and kidney disease. Mechanism of action: FG-2216 (YM-311; IOX-3; YM-311) inhibits HIF-PHDs (primarily PHD2), which normally hydroxylate proline residues in HIF-α subunits (HIF-1α, HIF-2α) under normoxia, targeting them for proteasomal degradation. Inhibition of PHDs stabilizes HIF-α, which heterodimerizes with HIF-1β and translocates to the nucleus, activating transcription of target genes including EPO (promotes erythropoiesis) and vascular endothelial growth factor (VEGF, supports renal vasculature) [1,3] - Therapeutic potential: FG-2216 (YM-311; IOX-3; YM-311) is a potential oral therapy for anemia associated with chronic kidney disease (CKD)/end-stage renal disease (ESRD), as it stimulates endogenous EPO production (avoiding risks of exogenous EPO, such as hypertension or thromboembolism). It also shows modest induction of HbF, which may benefit patients with sickle cell anemia or β-thalassemia [2,3] - Clinical relevance in ESRD: In ESRD, reduced renal PHD activity (due to hypoxia) is insufficient to compensate for impaired EPO synthesis. FG-2216 (YM-311; IOX-3; YM-311) enhances HIF stabilization in residual renal tissue, restoring EPO production and reversing anemia without relying on exogenous EPO administration [3] |
| 分子式 |
C12H9CLN2O4
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|---|---|
| 分子量 |
280.6639
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| 精确质量 |
280.025
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| 元素分析 |
C, 51.35; H, 3.23; Cl, 12.63; N, 9.98; O, 22.80
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| CAS号 |
223387-75-5
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| 相关CAS号 |
223387-75-5
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| PubChem CID |
6914666
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| 外观&性状 |
White to off-white solid powder
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| LogP |
1.799
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| tPSA |
99.52
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| 氢键供体(HBD)数目 |
3
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| 氢键受体(HBA)数目 |
5
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| 可旋转键数目(RBC) |
3
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| 重原子数目 |
19
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| 分子复杂度/Complexity |
366
|
| 定义原子立体中心数目 |
0
|
| SMILES |
ClC1C2=C([H])C([H])=C([H])C([H])=C2C(=C(C(N([H])C([H])([H])C(=O)O[H])=O)N=1)O[H]
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| InChi Key |
OUQVKRKGTAUJQA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C12H9ClN2O4/c13-11-7-4-2-1-3-6(7)10(18)9(15-11)12(19)14-5-8(16)17/h1-4,18H,5H2,(H,14,19)(H,16,17)
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| 化学名 |
2-[(1-chloro-4-hydroxyisoquinoline-3-carbonyl)amino]acetic acid
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| 别名 |
IOX3;IOX 3;IOX-3;FG-2216; FG 2216; FG2216; YM311; YM-311; YM 311.
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| HS Tariff Code |
2934.99.9001
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| 存储方式 |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| 运输条件 |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| 溶解度 (体外实验) |
DMSO: 56 mg/mL (199.5 mM)
Water:<1 mg/mL
Ethanol:<1 mg/mL
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|---|---|
| 溶解度 (体内实验) |
配方 1 中的溶解度: 2.5 mg/mL (8.91 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加,逐一添加), 悬浮液;超声助溶。
例如,若需制备1 mL的工作液,可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中,混匀;然后向上述溶液中加入50 μL Tween-80,混匀;加入450 μL生理盐水定容至1 mL。 *生理盐水的制备:将 0.9 g 氯化钠溶解在 100 mL ddH₂O中,得到澄清溶液。 配方 2 中的溶解度: ≥ 2.5 mg/mL (8.91 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加,逐一添加), 澄清溶液。 例如,若需制备1 mL的工作液,可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。 请根据您的实验动物和给药方式选择适当的溶解配方/方案: 1、请先配制澄清的储备液(如:用DMSO配置50 或 100 mg/mL母液(储备液)); 2、取适量母液,按从左到右的顺序依次添加助溶剂,澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明,并不一定代表此产品 的实际溶解配方): 10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline); 假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中,混合均匀/澄清;向上述体系中加入50 μL Tween-80,混合均匀/澄清;然后继续加入450 μL ddH2O (或 saline)定容至 1 mL; 3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例; 4、 如产品在配制过程中出现沉淀/析出,可通过加热(≤50℃)或超声的方式助溶; 5、为保证最佳实验结果,工作液请现配现用! 6、如不确定怎么将母液配置成体内动物实验的工作液,请查看说明书或联系我们; 7、 以上所有助溶剂都可在 Invivochem.cn网站购买。 |
| 制备储备液 | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5630 mL | 17.8152 mL | 35.6303 mL | |
| 5 mM | 0.7126 mL | 3.5630 mL | 7.1261 mL | |
| 10 mM | 0.3563 mL | 1.7815 mL | 3.5630 mL |
1、根据实验需要选择合适的溶剂配制储备液 (母液):对于大多数产品,InvivoChem推荐用DMSO配置母液 (比如:5、10、20mM或者10、20、50 mg/mL浓度),个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;
2、如果您找不到您想要的溶解度信息,或者很难将产品溶解在溶液中,请联系我们;
3、建议使用下列计算器进行相关计算(摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等);
4、母液配好之后,将其分装到常规用量,并储存在-20°C或-80°C,尽量减少反复冻融循环。
计算结果:
工作液浓度: mg/mL;
DMSO母液配制方法: mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL)。如该浓度超过该批次药物DMSO溶解度,请首先与我们联系。
体内配方配制方法:取 μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL ddH2O,混匀澄清。
(1) 请确保溶液澄清之后,再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶;
(2) 一定要按顺序加入溶剂 (助溶剂) 。
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT00456053 | Completed | Drug: FG2216 | Renal Anemia | FibroGen | December 2005 | Phase 2 |
Epo-induction in rhesus macques. (A) Circulating plasma Epo levels after a single oral dose of FG-2216.Blood.2007 Sep 15;110(6):2140-7. td> |
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Hemoglobin increases by PHI. (A) Hemoglobin (g/dL) values are presented with respect to number of days on treatment in 1 rhesus macaque treated with FG-2216 (●) and 1 control animal (○). (B) Hemoglobin levels in treated and control rhesus macaques.Blood.2007 Sep 15;110(6):2140-7. td> |