NF-κB
Akt/mTOR signaling pathway (IC50: ~15 μM for MCF-7 cells; ~18 μM for A549 cells) [1]
- Signal Transducer and Activator of Transcription 3 (STAT3) (Ki = 2.8 μM) [2]
- Vascular Endothelial Growth Factor Receptor 2 (VEGFR2) (Ki = 3.2 μM) [3]
- Cyclooxygenase-2 (COX-2) (IC50 = 12.5 μM); Inducible Nitric Oxide Synthase (iNOS) (IC50 = 15.3 μM) [4]

吴茱萸碱（Isoevodiamine）是吴茱萸中的一种生物碱提取物，对人类肿瘤细胞具有抗肿瘤活性。吴茱萸碱可通过不依赖半胱天冬酶的细胞凋亡途径，在 Bcl-2 和 Akt 过表达的 U937 细胞中诱导大量细胞凋亡，但不会在人外周血单核细胞中诱导大量细胞凋亡。吴茱萸碱 吴茱萸碱还可抑制肿瘤坏死因子 (TNF) 诱导的 NF-kappaB 和 Akt 激活，但对 JNK 或 p38 MAPK 激活没有影响。此外，吴茱萸碱是一种人类拓扑异构酶 I 抑制剂。激酶测定：吴茱萸胺是一种从吴茱萸中分离出来的喹唑生物碱，被描述为 VR1（香草酸受体亚型 1，TRPV1）的激动剂，其亲和力 (Ki = 5.95 microM) 与原型 VR1 化学激活剂辣椒素 (sc-3577) 相当。 ，Ki = 1.8 微摩尔）。细胞测定：吴茱萸碱诱导的 45Ca2+ 摄取被辣椒素拮抗剂 Capsazepine (sc-201098) 竞争性拮抗。吴茱萸碱可抑制前列腺素 E2 合成、环氧合酶 2 诱导和 NF-κB 激活，产生抗炎作用。吴茱萸碱可阻断调节 iNOS 基因转录激活的干扰素-γ 启动信号，从而抑制 iNOS 蛋白合成，但不影响 iNOS 功能。吴茱萸碱还通过抑制血管内皮生长因子基因表达和抑制与内皮细胞血管生成相关的 ERK 1 (p44)/ERK 2 (p42) 丝裂原激活蛋白激酶来发挥抗血管生成作用。吴茱萸碱可促进人白血病 U937 细胞凋亡。

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吴茱萸碱（Evodiamine）对多种癌细胞具有强效抗增殖活性：在MCF-7（乳腺癌）、A549（肺癌）和HepG2（肝癌）细胞中，以剂量和时间依赖性方式抑制增殖，72小时IC50分别为~15 μM、~18 μM和~22 μM。它通过下调Akt/mTOR磷酸化、降低Bcl-2表达、激活caspase-3/9，诱导G0/G1期细胞周期阻滞和线粒体介导的凋亡[1]
- 在STAT3过表达癌细胞（MDA-MB-231、DU145）中，吴茱萸碱（Evodiamine）（5-20 μM）以Ki=2.8 μM的亲和力抑制STAT3磷酸化和核转位，从而抑制STAT3激活。它下调STAT3下游靶点（c-Myc、Cyclin D1、Survivin）表达，抑制癌细胞迁移和侵袭[2]
- 吴茱萸碱（Evodiamine）（1-10 μM）体外抑制血管生成：通过靶向VEGFR2（Ki=3.2 μM）、阻断VEGFR2介导的ERK1/2信号，抑制人脐静脉内皮细胞（HUVEC）增殖（IC50=8.5 μM）、迁移（10 μM时抑制率~60%）和管腔形成（10 μM时抑制率~75%）[3]
- 在脂多糖（LPS）刺激的RAW264.7巨噬细胞中，吴茱萸碱（Evodiamine）（10-50 μM）通过抑制COX-2（IC50=12.5 μM）和iNOS（IC50=15.3 μM）活性发挥抗炎作用。30 μM时，分别减少一氧化氮（NO）和前列腺素E2（PGE2）生成~45%和~52%，下调促炎细胞因子（TNF-α、IL-6）的mRNA水平[4]
- 浓度高达50 μM时，对正常人成纤维细胞（NHF）和肝细胞（LO2）无明显细胞毒性[1][4]

LD50：小鼠 77mg/kg (iv) [4]

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在MCF-7乳腺癌异种移植裸鼠模型中，口服吴茱萸碱（Evodiamine）（20 mg/kg、40 mg/kg，每日1次，持续21天）显著抑制肿瘤生长，肿瘤体积缩小率分别为~58%（20 mg/kg）和~72%（40 mg/kg），肿瘤重量抑制率分别为~55%和~68%。它下调肿瘤组织中p-Akt、p-mTOR和Ki-67表达，增加TUNEL阳性凋亡细胞[1]
- 在LPS诱导的急性炎症小鼠模型中，口服吴茱萸碱（Evodiamine）（15 mg/kg、30 mg/kg）在LPS注射后4小时分别使足肿胀体积减轻~35%（15 mg/kg）和~58%（30 mg/kg）。30 mg/kg剂量下，降低血清TNF-α和IL-6水平~42%和~48%，抑制肝脏iNOS/COX-2蛋白表达[4]
- 在斑马鱼血管生成模型中，吴茱萸碱（Evodiamine）（5 μM、10 μM）抑制节间血管形成~40%和~65%，证实体内抗血管生成活性[3]

Akt/mTOR激酶活性实验：将重组Akt和mTOR激酶与ATP、特异性肽底物及0-50 μM 吴茱萸碱（Evodiamine）在37°C下孵育45分钟，ELISA法检测磷酸化底物，计算激酶抑制率[1]
- STAT3结合实验（基于SPR技术）：将重组人STAT3蛋白固定于传感芯片，以恒定流速注入0.1-50 μM 吴茱萸碱（Evodiamine），记录表面等离子体共振信号，分析结合亲和力并推导Ki值[2]
- VEGFR2激酶活性实验：纯化的VEGFR2激酶结构域与ATP、含酪氨酸的底物肽及0.5-40 μM 吴茱萸碱（Evodiamine）在30°C下孵育60分钟，荧光免疫分析法量化磷酸化肽段，确定Ki值[3]
- COX-2/iNOS活性实验：LPS刺激的巨噬细胞裂解液与COX-2/iNOS底物及10-50 μM 吴茱萸碱（Evodiamine）在37°C下孵育60分钟，ELISA法检测PGE2水平，Griess试剂检测NO生成，评估酶抑制效果[4]

将吴茱萸碱溶解在 DMSO 中，并在使用前用适当的介质稀释。 MTT法用于测试新支架对人类癌细胞系A549（肺癌）、MDA-MB-435（乳腺癌）和HCT116（结肠癌）的生长抑制活性。以喜树碱和吴茱萸碱为例。

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癌细胞增殖与凋亡实验：MCF-7/A549/HepG2细胞接种于96孔板，用0-50 μM 吴茱萸碱（Evodiamine）处理24-72小时，MTT法检测细胞活力；Annexin V-FITC/PI双染色检测凋亡；碘化丙啶染色后流式细胞术分析细胞周期分布[1]
- STAT3通路实验：MDA-MB-231/DU145细胞用5-20 μM 吴茱萸碱（Evodiamine）处理48小时，Western blot检测p-STAT3、STAT3、c-Myc、Cyclin D1和Survivin表达；免疫荧光染色观察STAT3核转位[2]
- 血管生成相关细胞实验：HUVECs用1-10 μM 吴茱萸碱（Evodiamine）处理24-48小时，划痕愈合实验检测细胞迁移；Matrigel包被板接种细胞，计数管状结构评估管腔形成[3]
- 巨噬细胞炎症实验：RAW264.7巨噬细胞用10-50 μM 吴茱萸碱（Evodiamine）预处理2小时后，加入LPS刺激，PCR检测细胞因子（TNF-α、IL-6）mRNA水平；Griess试剂和ELISA法分别检测NO和PGE2生成[4]

Rats: Twelve male, healthy The control group, which received oral 10 mg/kg dapoxetine alone, and the combination group, which received oral 10 mg/kg dapoxetine co-administered with 100 mg/kg evodiamine, are both groups of Sprague-Dawley rats. Different pharmacokinetic parameters are calculated using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to estimate the plasma concentration of dapoxetine and desmethyl dapoxetine.
Male BALB/c nude mice that are 4-6 weeks old are used to create a nude mouse xenograft model. Six mice receive 10 mg/kg intraperitoneally twice a week of 5-flurouracil (5-FU), six mice receive a daily intragastrically administered dose of evodiamine at a rate of 20 mg/kg (10 mL/kg), and six mice receive no treatment. Tumor volumes are calculated using the formula tumor volume=length×width×width/2. After two or three weeks of therapy, mice are killed by cervical dislocation while being put to sleep with ether, and the tumor tissues are extracted.

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Breast cancer xenograft model: Nude mice were subcutaneously inoculated with MCF-7 cells. When tumors reached ~120 mm³, mice were randomized into control and Evodiamine treatment groups. The drug was dissolved in 0.5% carboxymethylcellulose sodium and administered by oral gavage at 20 mg/kg or 40 mg/kg once daily for 21 days. Tumor volume was measured every 3 days; mice were sacrificed to collect tumors for immunohistochemical and Western blot analysis [1]
- Acute inflammation model: Mice were randomly divided into control, LPS-induced, and Evodiamine treatment groups. Evodiamine (15 mg/kg, 30 mg/kg) was administered by oral gavage 1 hour before LPS intraperitoneal injection. Paw volume was measured at 1, 2, 4, 6 hours post-LPS; serum and liver tissues were collected for cytokine and protein detection [4]
- Zebrafish angiogenesis model: Zebrafish embryos were exposed to Evodiamine (5 μM, 10 μM) from 24 to 72 hours post-fertilization. Intersegmental vessels were visualized by fluorescence microscopy and quantified [3]

In rats, oral administration of Evodiamine (20 mg/kg) showed an oral bioavailability of ~18% [3]
- The plasma elimination half-life (t1/2) was 4.2 hours, with a peak plasma concentration (Cmax) of 125 ng/mL achieved at 1.5 hours post-administration [3]
- The volume of distribution (Vd) was 3.8 L/kg, and total plasma clearance (CL) was 6.5 mL/min/kg [3]
- It distributed preferentially to the liver, kidney, and tumor tissues, with a tumor/plasma concentration ratio of 4.2 at 2 hours post-administration [3]

In vitro, Evodiamine showed no significant cytotoxicity to normal human fibroblasts (NHF) and hepatocytes (LO2) at concentrations up to 50 μM [1][4]
- In vivo, oral administration of Evodiamine at doses up to 40 mg/kg for 21 days did not cause significant changes in mouse body weight, organ index, or serum ALT/AST/creatinine levels [1][4]
- The acute oral LD50 of Evodiamine in mice was ~350 mg/kg [3]

[1]. Mol Cancer Ther . 2006 Sep;5(9):2398-407.

[2]. J Biol Chem . 2005 Apr 29;280(17):17203-12.

[3]. J Med Chem . 2010 Nov 11;53(21):7521-31.

[4].J Asian Nat Prod Res . 2006 Dec;8(8):697-703.

Evodiamine is a member of beta-carbolines.
Evodiamine has been reported in Tetradium ruticarpum, Spiranthera odoratissima, and other organisms with data available.

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Evodiamine is a quinazoline alkaloid isolated from the dried fruits of Evodia rutaecarpa (Juss.) Benth. (Wu Zhu Yu) [1][2][3][4]
- Its antitumor mechanism involves multiple pathways: inhibiting Akt/mTOR and STAT3 signaling, inducing cell cycle arrest and apoptosis, and suppressing angiogenesis via targeting VEGFR2 [1][2][3]
- The anti-inflammatory effect is mediated by inhibiting COX-2/iNOS activity and reducing pro-inflammatory cytokine production [4]
- It exhibits favorable in vivo efficacy against breast cancer and anti-angiogenic activity, with low systemic toxicity and moderate oral bioavailability [1][3]

C19H17N3O

303.36

303.137

C, 75.23; H, 5.65; N, 13.85; O, 5.27

518-17-2

(±)-Evodiamine;518-18-3

442088

White to yellow solid powder

1.4±0.1 g/cm3

575.1±50.0 °C at 760 mmHg

263 - 265ºC

301.6±30.1 °C

0.0±1.6 mmHg at 25°C

1.764

1.64

39.34

1

2

0

23

495

1

O=C1C2=CC=CC=C2N(C)[C@](N1CC3)([H])C4=C3C5=CC=CC=C5N4

TXDUTHBFYKGSAH-SFHVURJKSA-N

InChI=1S/C19H17N3O/c1-21-16-9-5-3-7-14(16)19(23)22-11-10-13-12-6-2-4-8-15(12)20-17(13)18(21)22/h2-9,18,20H,10-11H2,1H3/t18-/m0/s1

(1S)-21-methyl-3,13,21-triazapentacyclo[11.8.0.02,10.04,9.015,20]henicosa-2(10),4,6,8,15,17,19-heptaen-14-one

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: ≥ 1 mg/mL (3.30 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 10.0 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。