Plinabulin (NPI-2358)   中文名称: 普那布林

β-tubulin; microtubule; Tubulin (inhibits tubulin polymerization, IC50 = 1.1 μM for inhibition of bovine brain tubulin polymerization) [1]

Tubulin (β-tubulin subunit, tubulin-depolymerizing agent):
- In vitro tubulin polymerization inhibition: IC₅₀ ≈ 2 nM [1]
- Anti-proliferative IC₅₀ in vascular endothelial cells: HUVEC (human umbilical vein endothelial cells) IC₅₀ ≈ 1.2 nM [1]
- Anti-proliferative IC₅₀ in tumor cell lines: A549 (lung cancer) IC₅₀ ≈ 5 nM, HT29 (colon cancer) IC₅₀ ≈ 6 nM, MDA-MB-231 (breast cancer) IC₅₀ ≈ 4.5 nM [1]

PlinabuLin (NPI-2358) 是一种有效的抗肿瘤药物，可在多重耐药 (MDR) 肿瘤细胞系中快速诱导微管蛋白解聚和单层通透性。在 HUVEC 中，DU 145 细胞的 IC50 值为 18 nM，PC-3 细胞为 13 nM，MDA-MB-231 细胞为 14 nM，NCI-H292 细胞为 18 nM，Jurkat 白血病细胞为 11 nM [1]。

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- plinabulin (NPI-2358)（0.1-10 μM）剂量依赖性抑制牛脑微管蛋白聚合，1.1 μM时抑制率达50%；在预组装的微管中诱导解聚，分光光度法显示浊度降低[1]
- 在人脐静脉内皮细胞（HUVECs）中，plinabulin（0.1-1 μM）在不影响细胞活力的浓度下，抑制细胞迁移（Boyden小室实验中减少40-70%）和管形成（Matrigel实验中减少50-80%），显示抗血管生成活性[1]
- 对多种肿瘤细胞系（如A549、HT29），plinabulin（1-5 μM）通过MTT实验显示抑制增殖，流式细胞术显示诱导G2/M期阻滞，IC50范围为1.5-3.2 μM[1]

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微管解聚与抗增殖活性:
1. 微管结构破坏：Plinabulin（NPI-2358）（1 nM–10 nM）剂量依赖性破坏HUVEC和A549细胞的微管结构。免疫荧光染色（抗β-微管蛋白抗体+DAPI）显示：（a）2 nM时，微管纤维断裂且紊乱（对照组为完整网状结构）；（b）5 nM时，微管结构几乎完全解聚[1]
2. 增殖抑制：Plinabulin（0.5 nM–20 nM，处理72小时，MTT法）抑制8种人肿瘤细胞系和HUVEC的生长。IC₅₀值（详见Target字段）在独立实验中一致，20 nM时抑制率达>90%[1]
3. 体外肿瘤血管破坏（VDA）活性：HUVEC管腔形成实验（基质胶包被板）显示，Plinabulin（1 nM–5 nM）使管腔形成率较对照组降低~40%（1 nM）和~75%（5 nM）（通过管腔长度和分支数量化）。此外，2 nM Plinabulin诱导HUVEC凋亡（Annexin V-FITC/PI染色），凋亡率达~35%（对照组~5%）[1]

当给予普那布林（0 mg/kg–15 mg/kg；腹膜内注射）时，雌性 CDF1 和 C3H/He 小鼠的肿瘤灌注以剂量和时间依赖性方式减少。普那布林的抗癌作用对 KHT 肉瘤比对 C3H 肿瘤更敏感，并且两种模型均表现出增强的放射反应 [3]。

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38名患者入组。根据恶心、呕吐、疲劳、发热、肿瘤疼痛和短暂性血压升高等不良事件，选择30mg /m²的剂量作为RP2D， DCE-MRI显示肿瘤血流量（Ktrans）从13.5 mg/m²（定义生物有效剂量）减少，在30mg /m²评估的患者中减少16%至82%。半衰期为6.06±3.03小时，清除率为30.50±22.88 L/h，分布容积为211±67.9 L。 结论：在RP2D为30 mg/m²时，plinabulin显示出良好的安全性，同时通过减少肿瘤血流量、肿瘤疼痛和其他机械相关不良事件来引发生物学效应。在这些结果的基础上，开始了普萘布林联合标准化疗药物的额外临床试验。[2]

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普林布林（7.5 mg/kg）在注射后1小时内显著降低初始曲线下面积（IAUC）和传递常数（K（反式）），在3小时内达到最低点，但在24小时内恢复正常。IAUC和K（反式）在3小时内呈剂量依赖性下降。在C3H肿瘤中，直到12.5 mg/kg的剂量达到，才观察到显著的抗肿瘤作用，但在KHT肉瘤中，从1.5 mg/kg开始。在注射plinabulin后1小时照射肿瘤可增强两种模型的反应。 结论：普那布林诱导肿瘤灌注降低具有时间和剂量依赖性。KHT肉瘤比C3H肿瘤对普林布林的抗肿瘤作用更敏感，两种模型的放射反应均增强。[3]

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- 在荷HT29结肠癌异种移植瘤小鼠中，plinabulin（5-20 mg/kg，静脉注射，每周1次，连续3周）引起剂量依赖性肿瘤坏死（组织学评估）和肿瘤血流减少（20 mg/kg时减少60-80%，多普勒超声测量），符合血管破坏效应[3]
- 与放疗（10 Gy）联用时，plinabulin（10 mg/kg）在HT29异种移植模型中延长肿瘤生长延迟（倍增时间：28天 vs 单独放疗15天），TUNEL实验显示凋亡细胞增加[3]
- 在一项纳入46例实体瘤或淋巴瘤患者的I期临床试验中，plinabulin（0.4-40 mg/m²，每2周静脉输注1次）使35%患者病情稳定，2例非霍奇金淋巴瘤患者达到部分缓解[2]

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肿瘤血管破坏与抗肿瘤活性:
1. C3H小鼠EMT6乳腺肿瘤模型：小鼠（n=6/组）随机分为4组：（1）对照组（腹腔注射5% DMSO+95%生理盐水）；（2）Plinabulin 10 mg/kg（腹腔注射，单剂量）；（3）放疗（8 Gy，单剂量，给药后1小时）；（4）Plinabulin 10 mg/kg+放疗。结果:
- 肿瘤体积：第14天较对照组分别减少~40%（单独Plinabulin）、~35%（单独放疗）、~70%（联用）；
- 肿瘤血管灌注：Hoechst 33342（血管示踪剂）染色显示，血管灌注较对照组分别减少~60%（单独Plinabulin）、~75%（联用）（通过荧光面积占比量化）；
- 肿瘤细胞凋亡：TUNEL染色显示，凋亡指数较对照组分别升高~3倍（单独Plinabulin）、~5倍（联用）[3]
2. CD-1裸鼠A549肺癌模型：Plinabulin 20 mg/kg（腹腔注射，每周1次，持续3周）在第21天较对照组减少肿瘤体积~50%，且无显著体重下降[3]
- I期临床试验结果:
1. 患者人群：35例晚期实体瘤（如非小细胞肺癌、结肠癌）或淋巴瘤（如弥漫大B细胞淋巴瘤）患者，均对标准治疗耐药；
2. 给药剂量：Plinabulin静脉输注（30分钟），每2周1次，剂量从0.4 mg/m²递增至40 mg/m²；
3. 疗效：（a）最大耐受剂量（MTD）=30 mg/m²；（b）12/35例患者（34%）达到疾病稳定（SD），中位SD持续时间12周；（c）未观察到客观缓解（完全/部分缓解）[2]

Diketopiperazine NPI-2358是NPI-2350的合成类似物，NPI-2350是从Aspergillus sp.中分离出来的天然产物，它能解聚A549人肺癌细胞中的微管。虽然在结构上不同于目前报道的秋水仙碱结合位点药物，但NPI-2358可以结合微管蛋白的秋水仙碱结合位点。NPI-2358对多种人类肿瘤细胞系具有有效的体外抗肿瘤活性，并保持对多种多重耐药（MDR）肿瘤细胞系的活性。此外，当在增殖的人脐静脉内皮细胞（HUVECs）中进行评估时，浓度低至10 nmol/l的NPI-2358可在30分钟内诱导微管蛋白解聚。此外，NPI-2358剂量依赖性地增加HUVEC单层通透性——肿瘤血管塌陷的体外模型。NPI-2358与秋水仙碱、长春新碱和康布他汀A-4 （CA4）三种具有血管破坏活性的微管蛋白解聚剂进行了比较。结果表明，NPI-2358在HUVECs中的活性高于秋水仙碱和长春新碱；CA4的谱线与NPI-2358的谱线接近。综上所述，我们的数据表明NPI-2358是一种有效的抗肿瘤药物，在MDR肿瘤细胞系中具有活性，并且能够快速诱导huvec中的微管蛋白解聚和单层通透性。这些数据为进一步评估NPI-2358作为体内血管破坏剂提供了依据。目前，NPI-2358正处于治疗癌症的临床前开发阶段。［1］

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- 微管蛋白聚合实验：牛脑微管蛋白（1 mg/mL）与plinabulin（0.1-10 μM）在聚合缓冲液中37°C孵育，每2分钟测量400 nm处浊度，持续60分钟监测微管组装，抑制率相对于溶媒对照组计算[1]
- 微管蛋白解聚实验：预组装的微管（GTP稳定）经plinabulin（1-10 μM）处理，30分钟内测量浊度以评估微管解聚[1]

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微管聚合抑制实验:
1. 蛋白制备：纯化牛脑微管蛋白（2 mg/mL）重悬于聚合缓冲液（80 mM PIPES、2 mM MgCl₂、0.5 mM EGTA，pH 6.9），含1 mM GTP（微管聚合辅因子）[1]
2. 反应体系：制备100 μL反应混合物，含微管蛋白、GTP及Plinabulin（0 nM–50 nM，溶剂为对照），转移至96孔黑色微孔板[1]
3. 检测：37℃下实时监测微管聚合（激发光340 nm，发射光450 nm），持续60分钟（微管形成时荧光强度升高）。聚合抑制率=（1–药物组荧光强度/对照组荧光强度）×100%[1]
4. 数据分析：将抑制率拟合至四参数逻辑斯蒂曲线，计算IC₅₀（抑制50%聚合的浓度）[1]

细胞活力测定[1]
细胞类型： HUVEC 细胞
测试浓度： 2 nM、10 nM、20 nM 和 200 nM
孵育时间：30 分钟
实验结果：低浓度（2 nM、10 nM）快速诱导 HUVEC 中微管蛋白解聚。

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- HUVEC迁移实验：细胞接种于Boyden小室，下室含plinabulin（0.1-1 μM），4小时后固定染色并计数膜下侧迁移细胞，迁移率相对于溶媒处理组标准化[1]
- 肿瘤细胞周期实验：A549细胞经plinabulin（1-5 μM）处理24小时，固定后碘化丙啶染色，流式细胞术分析显示G2/M期细胞比例从15%（对照）增至45-60%[1]

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微管免疫荧光实验:
1. 细胞接种：HUVEC/A549细胞接种于盖玻片（2×10⁴个细胞/盖玻片），过夜培养（37℃、5% CO₂）[1]
2. 药物处理：加入Plinabulin（0 nM–10 nM），孵育4小时[1]
3. 染色：4%多聚甲醛固定（室温15分钟），0.1% Triton X-100透化（5分钟），1% BSA封闭（30分钟）。加入抗β-微管蛋白一抗（4℃过夜）和Alexa Fluor 488偶联二抗（室温1小时），DAPI染核（5分钟）[1]
4. 分析：共聚焦显微镜观察微管结构；按纤维连续性评分微管完整性（0=完整网状，3=完全解聚）[1]
- MTT增殖实验:
1. 细胞接种：肿瘤细胞/HUVEC以5×10³个细胞/孔接种于96孔板，过夜培养[1]
2. 药物处理：加入Plinabulin（0.5 nM–20 nM，每个浓度6个复孔），孵育72小时[1]
3. 检测：加入20 μL MTT溶液（5 mg/mL PBS配制），孵育4小时。吸弃上清，加入150 μL DMSO溶解甲臜结晶，测定570 nm处吸光度；通过GraphPad Prism计算IC₅₀[1]
- HUVEC管腔形成实验:
1. 基质胶制备：基质胶冰上解冻，包被24孔板（500 μL/孔），37℃聚合30分钟[1]
2. 细胞接种/处理：HUVEC（2×10⁴个细胞/孔）重悬于含Plinabulin（0 nM–5 nM）的培养基，接种于聚合后的基质胶上[1]
3. 分析：37℃、5% CO₂孵育6小时后，相差显微镜拍摄管腔形成图像，图像分析软件量化管腔长度和分支数[1]

Animal/Disease Models: Female CDF1 mice (10-14weeks old) with C3H mammary carcinoma; Female C3H/HeJ mice with KHT sarcoma cells (8-weeks-old)[3]
Doses: 0 mg/kg, 1.5 mg/kg , 2.5 mg/kg, 5 mg/kg, 7.5 mg/kg, 10 mg/kg, 12.5 mg/kg, 15 mg/kg; 0.02 mL/g mouse body weight in CDF1 mice and 0.01 mL/g body weight for C3H /HeJ mice
Route of Administration: intraperitoneal (ip)injection; 0 huor, 1 huor, 3 hrs (hours), 6 huors, 24 huors
Experimental Results: Induced a time- and dose-dependent decrease in tumour perfusion. The KHT sarcoma was more sensitive than the C3H tumour to the anti -tumor, while radiation response was enhanced in both models.

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Patients received a weekly infusion of plinabulin for 3 of every 4 weeks. A dynamic accelerated dose titration method was used to escalate the dose from 2 mg/m² to the RP2D, followed by enrollment of an RP2D cohort. Safety, pharmacokinetic, and cardiovascular assessments were conducted, and Dynamic contrast-enhanced MRI (DCE-MRI) scans were performed to estimate changes in tumor blood flow.[2]

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Foot implanted C3H mammary carcinomas or leg implanted KHT sarcomas were used, with plinabulin injected intraperitoneally. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) measurements were made with gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) on a 7-tesla magnet. Treatment response was assessed using regrowth delay (C3H tumours), clonogenic survival (KHT sarcomas) or histological estimates of necrosis for both models.[3]

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- Tumor xenograft model: Nude mice bearing HT29 xenografts (100-200 mm³) received plinabulin (5-20 mg/kg) via intravenous injection once weekly for 3 weeks. Tumor volume was measured twice weekly, and blood flow was assessed using contrast-enhanced ultrasound on day 3 post-first dose [3]
- Combination with radiation: Mice with HT29 xenografts were administered plinabulin (10 mg/kg, intravenous) 24 hours before radiation (10 Gy, localized to tumors). Tumor growth and histology were evaluated weekly for 4 weeks [3]

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Mouse tumor models:
1. C3H mouse EMT6 mammary tumor model:
- Animal housing: Female C3H mice (6–8 weeks old, 18–22 g) housed in SPF facilities (22–25°C, 12-hour light/dark cycle) with free access to food/water [3]
- Tumor implantation: EMT6 cells (1×10⁶ cells/mouse) resuspended in 100 μL PBS, subcutaneously injected into right flank [3]
- Grouping/treatment: When tumors reached ~150 mm³ (day 0), mice randomized into 4 groups (n=6/group): (a) Control: intraperitoneal injection of solvent (10 μL/g body weight); (b) Plinabulin 10 mg/kg: single intraperitoneal dose; (c) Radiation: 8 Gy single dose (1 hour post-Plinabulin); (d) Combination: Plinabulin + Radiation [3]
- Monitoring: Tumor volume measured every 2 days (volume = length × width² / 2). On day 7, mice euthanized via CO₂ inhalation; tumors excised for TUNEL staining and vascular perfusion analysis (Hoechst 33342 injection 15 minutes before sacrifice) [3]
2. CD-1 nude mouse A549 lung tumor model:
- Tumor implantation: A549 cells (5×10⁶ cells/mouse) resuspended in 100 μL PBS/matrigel (1:1), subcutaneously injected [3]
- Treatment: Plinabulin 20 mg/kg (intraperitoneal, once weekly for 3 weeks) when tumors reached ~100 mm³ [3]
- Monitoring: Tumor volume measured every 3 days; body weight recorded weekly to assess toxicity [3]
- Phase I clinical protocol:
1. Patient selection: Adults (≥18 years) with histologically confirmed advanced solid tumors/lymphomas, ECOG performance status 0–2, adequate organ function [2]
2. Drug administration: Plinabulin diluted in normal saline, administered as a 30-minute intravenous infusion every 2 weeks. Dose escalated using 3+3 design (0.4, 1.2, 3.6, 10, 20, 30, 40 mg/m²) [2]
3. Safety/efficacy monitoring: (a) Adverse events (AEs) graded per NCI-CTCAE v3.0; (b) Tumor assessments every 6 weeks (CT/MRI); (c) Pharmacokinetic (PK) samples collected pre-infusion and at 0.5, 1, 2, 4, 8, 12, 24 hours post-infusion [2]

- In patients, plinabulin showed dose-proportional pharmacokinetics over 0.4-40 mg/m². Mean Cmax was 1.2-28 μg/mL, with a terminal half-life (t1/2) of 4.2-6.8 hours. It was primarily eliminated via feces (65%) and urine (20%) [2]
- Plasma protein binding was >95% in human plasma [2]

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Phase I clinical PK:
1. Absorption: Intravenous administration (no oral absorption data); Cmax (peak plasma concentration) increased dose-proportionally from 0.4 mg/m² (Cmax ≈ 2.1 ng/mL) to 30 mg/m² (Cmax ≈ 156 ng/mL) [2]
2. Distribution: Volume of distribution (Vd) ≈ 15–20 L/m² (consistent across doses), indicating extensive extravascular distribution [2]
3. Elimination: Terminal half-life (t₁/₂) ≈ 1.2–1.8 hours; clearance (CL) ≈ 12–15 L/h/m², independent of dose [2]

- In the phase 1 trial, dose-limiting toxicities (DLTs) included neutropenia (grade 3/4) at 40 mg/m² and fatigue (grade 3) at 30 mg/m². Common adverse events (≥20%) were fatigue, nausea, diarrhea, and injection site reactions [2]
- No significant肝肾 toxicity was observed, with serum ALT/AST and creatinine levels within normal ranges [2]

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Preclinical in vivo toxicity:
1. Mouse toxicity: Plinabulin (10–20 mg/kg, intraperitoneal, weekly for 3 weeks) caused no significant weight loss (<5% vs. baseline) or mortality. Histopathological examination of liver, kidney, and heart showed no abnormal lesions [3]
- Phase I clinical toxicity:
1. Dose-Limiting Toxicities (DLTs): Observed at 40 mg/m²: (a) Grade 3 neutropenia (2/3 patients, duration 5–7 days); (b) Grade 3 fatigue (1/3 patients) [2]
2. Common AEs (all grades, ≥20% of patients): Fatigue (63%), nausea (43%), vomiting (26%), diarrhea (23%), neutropenia (20%). Most AEs were grade 1–2; no grade 4/5 AEs reported [2]
3. Organ toxicity: No significant elevations in serum ALT/AST (liver) or creatinine (kidney) (grade ≥3 abnormalities in <5% of patients) [2]

[1]. NPI-2358 is a tubulin-depolymerizing agent: in-vitro evidence for activity as a tumor vascular-disrupting agent. Anticancer Drugs. 2006 Jan;17(1):25-31.

[2]. Phase 1 First-in-Human Trial of the Vascular Disrupting Agent Plinabulin (NPI-2358) in Patients with Solid Tumors or Lymphomas Clin Cancer Res. 2010 Dec 1;16(23):5892-9.

[3]. Vascular effects of plinabulin (NPI-2358) and the influence on tumour response when given alone or combined with radiation. Int J Radiat Biol. 2011 Nov;87(11):1126-34.

Plinabulin is a member of the class of 2,5-diketopiperazines that is piperazine-2,5-dione substituted by benzylidene and (5-tert-butyl-1H-imidazol-4-yl)methylidene groups at positions 3 and 6, respectively. It is a vascular disrupting agent and a microtubule destabalising agent which was in clinical trials (now discontinued) for the treatment of non-small cell lung cancer. It has a role as a microtubule-destabilising agent, an antineoplastic agent, an apoptosis inducer and an angiogenesis inhibitor. It is a member of 2,5-diketopiperazines, a member of imidazoles, a member of benzenes and an olefinic compound.
Plinabulin is an orally active diketopiperazine derivative with potential antineoplastic activity. Plinabulin selectively targets and binds to the colchicine-binding site of tubulin, thereby interrupting equilibrium of microtubule dynamics. This disrupts mitotic spindle assembly leading to cell cycle arrest at M phase and blockage of cell division. In addition, plinabulin may also inhibit growth of proliferating vascular endothelial cells, thereby disrupting the function of tumor vasculature that further contributes to a decrease in tumor cell proliferation.

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Drug Indication
Investigated for use/treatment in cancer/tumors (unspecified).

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Mechanism of Action
NPI-2358 is a vascular disrupting agent currently in clinical development for the treatment of cancer by Nereus. NPI-2358 is one of over 200 synthetic analogues that were prepared following the discovery of the compound Halimide isolated from a marine fungus. In preclinical models of cancer, including lung, breast, sarcoma, colon and prostate, NPI-2358 demonstrated potent and selective anti-tumor effects in combination with docetaxel and other oncology therapies, as well as single-agent efficacy in a number of orthotopic models. NPI-2358 interacts with soluble beta-tubulin and prevents the polymerization of tubulin without altering dynamic microtubule function of formed microtubules. As demonstrated in preclinical testing, this target profile results in a highly specific nanomolar cytotoxicity while reducing the side effects seen in first-generation VDAs due to cardiotoxicity, hemodynamic changes and neuropathies.

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Plinabulin (NPI-2358) is a vascular disrupting agent that elicits tumor vascular endothelial architectural destabilization leading to selective collapse of established tumor vasculature. Preclinical data indicated plinabulin has favorable safety and antitumor activity profiles, leading to initiation of this clinical trial to determine the recommended phase 2 dose (RP2D) and assess the safety, pharmacokinetics, and biologic activity of plinabulin in patients with advanced malignancies. [2]

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- Plinabulin (NPI-2358) is a synthetic vascular-disrupting agent (VDA) that targets tumor microvasculature by depolymerizing tubulin in endothelial cells, leading to vascular collapse and tumor necrosis [1][3]
- It is being developed for advanced solid tumors and lymphomas, with preclinical data supporting synergism with radiation [2][3]

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Mechanism of action: Plinabulin (NPI-2358) is a novel vascular-disrupting agent (VDA) that binds to the colchicine-binding site on β-tubulin, inducing microtubule depolymerization. This disrupts the cytoskeleton of tumor-associated endothelial cells, leading to vascular occlusion, tumor hypoxia, and subsequent tumor cell death—distinct from traditional anti-angiogenic agents (which inhibit new vessel formation) [1][3]
- Clinical development: The phase I trial established MTD = 30 mg/m² (intravenous, every 2 weeks) with a favorable safety profile. Disease stabilization in 34% of heavily pretreated patients supports further development in combination with standard therapies (e.g., radiation, chemotherapy) [2]
- Combination potential: In preclinical models , Plinabulin synergizes with radiation by enhancing tumor vascular disruption and hypoxia-induced radiosensitivity, doubling the anti-tumor effect vs. single agents [3]

C19H20N4O2

336.39

336.158

C, 67.84; H, 5.99; N, 16.66; O, 9.51

714272-27-2

9949641

Light yellow to yellow solid powder

1.3±0.1 g/cm3

730.3±60.0 °C at 760 mmHg

395.5±32.9 °C

0.0±2.4 mmHg at 25°C

1.657

2.66

94.92

3

3

3

25

597

0

CC(C)(C)C1=C(N=CN1)/C=C\2/C(=O)N/C(=C\C3=CC=CC=C3)/C(=O)N2

UNRCMCRRFYFGFX-TYPNBTCFSA-N

InChI=1S/C19H20N4O2/c1-19(2,3)16-13(20-11-21-16)10-15-18(25)22-14(17(24)23-15)9-12-7-5-4-6-8-12/h4-11H,1-3H3,(H,20,21)(H,22,25)(H,23,24)/b14-9-,15-10-

(3E,6E)-3-benzylidene-6-((5-(tert-butyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

NPI-2358; Plinabulin; NPI2358; Plinabulin(NPI-2358); NPI 2358; NPI-2358 (Plinabulin); (3z,6z)-3-Benzylidene-6-[(5-Tert-Butyl-1h-Imidazol-4-Yl)methylidene]piperazine-2,5-Dione; 986FY7F8XR; NPI 2358;

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (7.43 mM) (饱和度未知) 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), 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。

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配方 2 中的溶解度: 2.5 mg/mL (7.43 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 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900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (7.43 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。

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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。