Ziv-aflibercept   中文名称: 阿柏西普

ziv-aflibercept binds VEGF and placental growth factor (PIGF) as a soluble decoy receptor (no IC50/Ki/EC50 data provided)[1]

Ziv-aflibercept（0.25、0.5、1.0 和 5 mg/mL；24 小时）以剂量依赖性方式降低 ARPE-19 细胞中的线粒体膜电位 [1]。

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ziv-aflibercept 在10×临床浓度（2 mg/4 mL玻璃体等效）下显著降低ARPE-19人视网膜色素上皮细胞活力至77.25% ± 2.1%（p < 0.0001），但在1/2×、1×或2×剂量下与对照组相比无统计学显著变化[1]

ziv-aflibercept 诱导线粒体毒性，在1×（73.50% ± 2.93%，p < 0.0001）、2×（64.83% ± 2.7%，p < 0.0001）和10×（49.65% ± 4.22%，p = 0.0002）浓度下降低线粒体膜电位（ΔΨm），表明早期凋亡；1/2×剂量（91.57% ± 2.54%）未观察到显著ΔΨm变化[1]

ziv-aflibercept 在10×浓度下增加渗透压至418 mOsm/kg（对照组为324 mOsm/kg），但1/2×、1×和2×剂量下的渗透压（分别为324、330和342 mOsm/kg）与对照组无显著差异[1]

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Ziv-aflibercept 在临床剂量（1×）下对人视网膜色素上皮细胞（ARPE-19系）暴露24小时后未降低细胞活力（基于Malik等研究）。[2]

在临床剂量两倍（2×）浓度下，其在Müller细胞（MIO-M1系）中显著降低活力，而雷珠单抗、阿柏西普或贝伐珠单抗无此效应，表明高浓度下存在细胞类型特异性毒性。[2]

在兔子的右眼中，ziv-aflibercept（25 mg/mL；眼内注射；单剂量）基本上不会诱发白内障、视网膜脱离或任何相关问题[2]。

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兔模型中，玻璃体内注射ziv-aflibercept（0.05 ml, 25 mg/ml）后1天和7天，眼底检查、光学相干断层扫描（OCT）和全视野视网膜电图（ERG）均未显示毒性；组织学和透射电镜证实无解剖损伤。[2]
人类案例报告（如新生血管性年龄相关性黄斑变性、糖尿病性黄斑水肿）显示，玻璃体内注射（1.25 mg/剂）后视力改善，视网膜内/下液减少，随访期间未见炎症、白内障进展或视网膜毒性。[2]

细胞活力测定[1]
细胞类型： ARPE-19 细胞。
测试浓度：0.25、0.5、1.0 和 5 mg/mL。
孵化持续时间：24小时。
实验结果：严重影响细胞活力（低于 1 mg/mL）。

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细胞活力实验中，ARPE-19细胞以5.0 × 10⁵细胞/孔接种于六孔板，加入2 mL培养基，标准条件（37°C、5% CO₂、95%湿度）孵育24小时，后用ziv-aflibercept在1/2×、1×、2×或10×临床浓度或对照（无药物）处理24小时。细胞经胰蛋白酶消化、离心（1000 rpm，5分钟），重悬于1 mL培养基，通过自动化台盼蓝染色排除法分析活细胞/死细胞；实验重复三次并独立重复两次[1]

线粒体膜电位（ΔΨm）实验中，ARPE-19细胞以1.0 × 10⁵细胞/孔接种于24孔板，过夜孵育，后用ziv-aflibercept处理24小时。JC-1染料检测ΔΨm：健康细胞（红色荧光，590 nm）对比凋亡/坏死细胞（绿色荧光，529 nm）。使用荧光成像扫描仪测量红绿荧光比值，实验重复四次并独立重复两次[1]

含ziv-aflibercept培养基的渗透压通过自动渗透压仪（基于冰点降低法）测量；所有浓度（1/2×至10×）样本均被测试[1]

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活力测试中，ARPE-19细胞暴露于ziv-aflibercept的1/2×、1×、2×和10×临床浓度（基于玻璃体分布）24小时，通过自动化台盼蓝排除法评估活力，结果归一化至对照组并进行统计分析。[2]
Müller细胞（MIO-M1系）实验采用低剂量（1/2×）和临床等效剂量；2×浓度下的活力降低通过标准化方法量化。[2]

Nine rabbits received unilateral intravitreal injections of 0.05 ml ziv-aflibercept (25 mg/ml). Assessments included baseline/post-injection serum/vitreous/aqueous osmolarity, funduscopy, OCT, ERG (days 1 and 7), and histopathological analysis. No sham controls were used; outcomes focused on absence of complications (e.g., cataract, retinal detachment).[2]

The half-life of ziv-aflibercept is 7.1 days, identical to aflibercept, based on structural similarity.[2]

strong>ziv-aflibercept exhibited mitochondrial toxicity at clinically relevant concentrations (1×, 2×, and 10×) in ARPE-19 cells, with reduced ΔΨm suggesting early apoptosis; cell viability decreased only at 10× dose[1]

Increased cytotoxicity may be attributed to lower pH and higher osmolality compared to aflibercept, though no systemic toxicokinetic data (e.g., plasma protein binding, organ toxicity) were evaluated[1]
No ocular or systemic toxicity was reported in rabbits or humans after intravitreal ziv-aflibercept administration. Serum and intraocular osmolarity remained unchanged post-injection.[2]
Mitochondrial toxicity observed in vitro at supraclinical doses (e.g., reduced ΔΨm in ARPE-19 cells), but no in vivo evidence of organ toxicity or drug interactions.[2]

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
This record refers to the use of ziv-aflibercept for treating cancer. No information is available on the use of ziv-aflibercept during breastfeeding. Because ziv-aflibercept is a large protein molecule with a molecular weight of 115,000, the amount in milk is likely to be very low and absorption is unlikely because it is probably destroyed in the infant's gastrointestinal tract. However, ziv-aflibercept is usually used in combination with other potentially toxic chemotherapy agents and most sources consider breastfeeding to be contraindicated during maternal antineoplastic drug therapy. Intravitreal aflibercept decreased vascular endothelial growth factor in breastmilk in one woman. Since VEGF is present in human milk and is thought to help in maturation of the infant’s gastrointestinal tract, concern has been raised about the maternal use of VEGF inhibitors during breastfeeding. Note that the typical alternative to breastmilk is infant formula, which contains no VEGF.

◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.

◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

[1]. Safety profiles of anti-VEGF drugs: bevacizumab, ranibizumab, aflibercept and ziv-aflibercept on human retinal pigment epithelium cells in culture. Br J Ophthalmol. 2014 Jun;98 Suppl 1(Suppl 1):i11-16.

[2]. Fusion proteins for treatment of retinal diseases: aflibercept, ziv-aflibercept, and conbercept. Int J Retina Vitreous. 2016 Feb 1;2:3.

Purpose: To compare the safety profiles of antivascular endothelial growth factor (VEGF) drugs ranibizumab, bevacizumab, aflibercept and ziv-aflibercept on retinal pigment epithelium cells in culture. Methods: Human retinal pigment epithelium cells (ARPE-19) were exposed for 24 h to four anti-VEGF drugs at 1/2×, 1×, 2× and 10× clinical concentrations. Cell viability and mitochondrial membrane potential assay were performed to evaluate early apoptotic changes and rate of overall cell death. Results: Cell viability decreased at 10× concentrations in bevacizumab (82.38%, p=0.0001), aflibercept (82.68%, p=0.0002) and ziv-aflibercept (77.25%, p<0.0001), but not at lower concentrations. However, no changes were seen in cell viability in ranibizumab-treated cells at all concentrations including 10×. Mitochondrial membrane potential was slightly decreased in 10× ranibizumab-treated cells (89.61%, p=0.0006) and 2× and 10× aflibercept-treated cells (88.76%, 81.46%; p<0.01, respectively). A larger reduction in mitochondrial membrane potential was seen at 1×, 2× and 10× concentrations of bevacizumab (86.53%, 74.38%, 66.67%; p<0.01) and ziv-aflibercept (73.50%, 64.83% and 49.65% p<0.01) suggestive of early apoptosis at lower doses, including the clinical doses. Conclusions: At clinical doses, neither ranibizumab nor aflibercept produced evidence of mitochondrial toxicity or cell death. However, bevacizumab and ziv-aflibercept showed mild mitochondrial toxicity at clinically relevant doses. [1]

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In the last few years, monoclonal antibodies have revolutionized the treatment of retinal neovascular diseases. More recently, a different class of drugs, fusion proteins, has provided an alternative treatment strategy with pharmacological differences. In addition to commercially available aflibercept, two other drugs, ziv-aflibercept and conbercept, have been studied in antiangiogenic treatment of ocular diseases. In this scenario, a critical review of the currently available data regarding fusion proteins in ophthalmic diseases may be a timely and important contribution. Aflibercept, previously known as VEGF Trap Eye, is a fusion protein of VEGF receptors 1 and 2 and a treatment for several retinal diseases related to angiogenesis. It has firmly joined ranibizumab and bevacizumab as an important therapeutic option in the management of neovascular AMD-, DME- and RVO-associated macular edema. Ziv-aflibercept, a systemic chemotherapeutic agent approved for the treatment of metastatic colorectal cancer, has recently drawn attention because of its potential for intravitreal administration, since it was not associated with ERG-related signs of toxicity in an experimental study and in human case reports. Conbercept is a soluble receptor decoy that blocks all isoforms of VEGF-A, VEGF-B, VEGF-C, and PlGF, which has a high binding affinity for VEGF and a long half-life in vitreous. It has been studied in a phase three clinical trial and has shown efficacy and safety. This review discusses three fusion proteins that have been studied in ophthalmology, aflibercept, ziv-aflibercept and conbercept, with emphasis on their clinical application for the treatment of retinal diseases.[2]

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Ziv-aflibercept is structurally identical to aflibercept but differs in excipients and osmolarity (1000 mOsm/l vs. aflibercept’s 286 mOsm/l). FDA-approved for metastatic colorectal cancer (2012), it is used off-label for retinal diseases due to lower cost. Clinical intravitreal dose is 1.25 mg (0.05 ml of 25 mg/ml solution).[2]

Preliminary data suggest efficacy in reducing macular edema and improving visual acuity in AMD/DME, though large-scale trials are needed to confirm safety and dosing.[2]

1609655-49-3

Colorless to light yellow liquid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
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示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
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注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
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注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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