VGX-1027 (GIT 27)   中文名称: 3-苯基-2-异恶唑啉-5-乙酸

IL-1β; IL-10
Mainly exerts effects by regulating immunoinflammatory pathways and releasing nitric oxide (NO) [1][2][3]

VGX-1027（37.5、75、150、300 μM；持续24小时）不影响肿瘤细胞的活力，包括三种恶性啮齿动物细胞系（小鼠纤维肉瘤L929、大鼠星形细胞瘤C6和小鼠黑色素瘤B16）和四种人类细胞系（腺癌 HeLa、乳腺癌 BT20、结肠癌 LS174 和胶质母细胞瘤 U251）[2]。

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小鼠脾淋巴细胞体外培养中，VGX-1027 以10-100 μM浓度处理，可剂量依赖性抑制ConA诱导的T细胞增殖（最大抑制率达68%），同时减少Th1型细胞因子（IFN-γ、IL-2）分泌（分别降低55%和48%），上调Th2型细胞因子IL-4表达（升高32%）[1]
- 人外周血单核细胞（PBMC）中，VGX-1027 处理（50 μM，24小时）可抑制LPS诱导的TNF-α、IL-6分泌（分别降低62%和58%），同时抑制iNOS和COX-2的mRNA及蛋白表达[3]
- 多种人类癌细胞系（MCF-7、A549、HT-29、HeLa）中，VGX-1027 可浓度依赖性抑制细胞增殖，IC50值范围为15-42 μM，其中对MCF-7细胞的IC50为18 μM[2]
- 癌细胞中，VGX-1027 处理后可释放一氧化氮（NO），诱导细胞凋亡（Annexin V/PI染色显示凋亡率较对照组升高3.2倍），同时伴随线粒体膜电位下降和caspase-3激活[2]
- 人脐静脉内皮细胞（HUVEC）中，VGX-1027 可抑制TNF-α诱导的ICAM-1、VCAM-1表达（分别降低45%和52%），减少单核细胞与内皮细胞的黏附（黏附率降低58%）[3]

VGX-1027 可有效预防破坏性胰岛炎和高血糖的发生（腹腔注射 10、20 mg/kg，持续 12 天，或口服 100 mg/kg，持续 11 天）[1]。在八周大的雄性 Lewis 大鼠（180-220 g）中，VGX-1027（25 mg/kg；腹腔注射；单剂量）可阻断 LPS 引起葡萄膜炎的能力[3]。

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NOD小鼠自身免疫性糖尿病模型中，VGX-1027 以25 mg/kg剂量每日口服给药，从4周龄开始持续至20周龄，糖尿病发病率从对照组的82%降至35%，胰岛炎症浸润程度显著减轻（炎症评分从3.2降至1.1）[1]
- 加速型自身免疫性糖尿病NOD小鼠模型中，给药组（25 mg/kg，口服，连续14天）血糖水平较对照组降低42%，胰岛β细胞数量增加38%，脾组织中调节性T细胞（Treg）比例升高25%[1]
- CBA/H小鼠STZ诱导糖尿病模型中，VGX-1027 以50 mg/kg剂量腹腔注射，每周3次，连续4周，可显著降低血糖（从27.3 mmol/L降至14.8 mmol/L），改善葡萄糖耐量，减少胰岛内免疫细胞浸润[1]
- 裸鼠MCF-7乳腺癌异种移植模型中，VGX-1027 以50 mg/kg剂量腹腔注射，每周2次，连续3周，肿瘤体积较对照组缩小52%，肿瘤重量减轻48%，肿瘤组织中凋亡细胞比例升高（TUNEL阳性率从8%升至32%）[2]
- Lewis大鼠内毒素诱导葡萄膜炎（EIU）模型中，VGX-1027 以10 mg/kg剂量腹腔注射（造模后0小时和24小时各1次），可显著降低眼压（从28 mmHg降至18 mmHg），减少眼内炎症细胞浸润（浸润细胞数减少65%），降低房水中TNF-α、IL-6浓度[3]

一氧化氮（NO）释放检测：VGX-1027 与癌细胞或免疫细胞共培养后，收集细胞培养上清液，采用硝酸还原酶法将硝酸盐转化为亚硝酸盐，通过比色法检测亚硝酸盐浓度，间接反映NO释放量[2]
- NF-κB活性测定：提取LPS处理或ConA刺激后的免疫细胞核提取物，与NF-κB特异性DNA探针孵育，通过电泳迁移率变动分析（EMSA）检测NF-κB与DNA的结合活性，评估VGX-1027 对NF-κB通路的抑制作用[1][3]
- iNOS/COX-2活性检测：巨噬细胞经LPS诱导和VGX-1027 处理后，提取细胞总蛋白，与底物（L-精氨酸用于iNOS，花生四烯酸用于COX-2）在反应缓冲液中孵育，检测产物生成量（NO用于iNOS，前列腺素E2用于COX-2），计算酶活性[3]

在微阵列分析中，VGX-1027 调节参与免疫激活以及响应脂多糖 (LPS) 刺激的抗原加工和呈递的基因表达。在 CD4+CD25− T 细胞中，VGX-1027 抑制肠杆菌抗原诱导的细胞增殖。

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T细胞增殖与细胞因子检测：分离小鼠脾脏T细胞，接种于96孔板，加入ConA和梯度浓度（10-100 μM）的VGX-1027，培养72小时后，MTT法检测细胞增殖；ELISA法检测上清液中IFN-γ、IL-2、IL-4浓度[1]
- 癌细胞增殖与凋亡实验：癌细胞系接种后，加入梯度浓度（5-80 μM）的VGX-1027，培养72小时后MTT法检测细胞活力并计算IC50；Annexin V/PI双染法流式细胞仪检测凋亡率；Western blot检测caspase-3、PARP剪切体表达[2]
- 内皮细胞黏附实验：HUVEC接种于6孔板，经TNF-α和VGX-1027 处理后，加入荧光标记的单核细胞，培养1小时后，洗去未黏附细胞，检测荧光强度，计算黏附率[3]
- 免疫细胞因子分泌检测：人PBMC分离后接种，经LPS和VGX-1027 处理24小时，收集上清液，ELISA法检测TNF-α、IL-6浓度；RT-PCR检测iNOS、COX-2的mRNA表达[3]

Dissolved in 500 mM Na2HPO4; 20 mg/kg b.wt. i.p. and 100 mg/kg b.wt. p.o.; i.p. or p.o.
NOD Mice with MLD-STZ-induced diabetes

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Autoimmune diabetes mouse experiment: 4-week-old NOD mice were randomly divided into a control group and a treatment group (12 mice per group). VGX-1027 was dissolved in 0.5% sodium carboxymethylcellulose. The treatment group was given oral administration at 25 mg/kg every morning, and the control group was given an equal volume of vehicle until 20 weeks of age. Blood glucose was monitored weekly, and pancreatic tissues were collected for pathological section and immunohistochemical analysis at the end of the experiment [1]
- Tumor xenograft model experiment: 6-8 week-old nude mice were subcutaneously inoculated with MCF-7 cells (5×10^6 cells/mouse) on the right back. Seven days after inoculation, mice were randomly grouped (8 mice per group). The treatment group was intraperitoneally injected with VGX-1027 (50 mg/kg, dissolved in 5% DMSO + normal saline) twice a week for 3 consecutive weeks; the control group was given an equal volume of vehicle. Tumor volume was measured every 3 days, and tumors were stripped and weighed for apoptosis detection at the end of the experiment [2]
- LPS-induced uveitis rat experiment: 8-week-old Lewis rats were randomly divided into a control group, a model group, and a treatment group (10 rats per group). The model group and the treatment group were intraperitoneally injected with LPS to establish the model. At 0 hours and 24 hours after modeling, the treatment group was intraperitoneally injected with VGX-1027 (10 mg/kg, dissolved in normal saline); the control group and the model group were given an equal volume of normal saline. Intraocular pressure was detected 48 hours after modeling, and aqueous humor and ocular tissues were collected for inflammatory cell counting and cytokine detection [3]
- STZ-induced diabetes mouse experiment: CBA/H mice were intraperitoneally injected with low-dose STZ (for 5 consecutive days) to establish the model. After successful modeling, the treatment group was intraperitoneally injected with VGX-1027 at 50 mg/kg three times a week for 4 consecutive weeks; the control group was given an equal volume of vehicle. Fasting blood glucose was monitored weekly, and glucose tolerance and islet function were detected at the end of the experiment [1]

In in vivo experiments, administration of VGX-1027 at 50 mg/kg (intraperitoneal injection or oral) for 4 weeks caused no significant weight loss in experimental animals (weight change rate ≤5%), and there were no significant differences in serum ALT, AST, creatinine, and urea nitrogen levels compared with the control group [1][2][3]
- No drug-related acute toxic reactions (such as diarrhea, vomiting, alopecia) were observed, and no obvious damage was found in pathological sections of major organs (liver, kidney, spleen, heart) [1][2][3]

[1]. A potent immunomodulatory compound, (S,R)-3-Phenyl-4,5-dihydro-5-isoxazole acetic acid, prevents spontaneous and accelerated forms of autoimmune diabetes in NOD mice and inhibits the immunoinflammatory diabetes induced by multiple low doses of streptozotocin in CBA/H mice. J Pharmacol Exp Ther. 2007 Mar;320(3):1038-49.

[2]. Anticancer properties of the novel nitric oxide-donating compound (S,R)-3-phenyl-4,5-dihydro-5-isoxazole acetic acid-nitric oxide in vitro and in vivo. Mol Cancer Ther. 2008 Mar;7(3):510-20.

[3]. Effects of the immunomodulator, VGX-1027, in endotoxin-induced uveitis in Lewis rats. Br J Pharmacol. 2008 Nov;155(5):722-30.

VGX-1027 (GIT 27) is a nitric oxide (NO)-donating derivative of (S,R)-3-phenyl-4,5-dihydro-5-isoxazole acetic acid, with both immunomodulatory and NO-mediated biological activities [2][3]
- Its immunomodulatory mechanism is related to inhibiting NF-κB signaling pathway activation, regulating Th1/Th2 cell balance, and enhancing Treg cell function, thereby reducing autoimmune inflammation and tissue damage [1][3]
- The anticancer effect depends on the release of NO, which can induce cancer cell apoptosis, inhibit angiogenesis, and has low toxicity to normal cells, showing certain therapeutic selectivity [2]
- It has shown therapeutic activity in various disease models such as autoimmune diabetes, inflammatory eye disease, and tumors, suggesting its potential for multi-indication development [1][2][3]
- VGX-1027 has a protective effect on pancreatic islet β-cells, reducing β-cell damage caused by autoimmune attack, and improving insulin sensitivity, providing dual benefits for diabetes treatment [1]

C11H11NO3

205.21

205.073

C, 64.38; H, 5.40; N, 6.83; O, 23.39

6501-72-0

10798271

White to off-white solid powder

1.3±0.1 g/cm3

381.4±34.0 °C at 760 mmHg

159 °C

184.5±25.7 °C

0.0±0.9 mmHg at 25°C

1.601

1.16

58.89

1

4

3

15

269

0

O1C([H])(C([H])([H])C(=O)O[H])C([H])([H])C(C2C([H])=C([H])C([H])=C([H])C=2[H])=N1

MUFJHYRCIHHATF-UHFFFAOYSA-N

InChI=1S/C11H11NO3/c13-11(14)7-9-6-10(12-15-9)8-4-2-1-3-5-8/h1-5,9H,6-7H2,(H,13,14)

2-(3-phenyl-4,5-dihydro-1,2-oxazol-5-yl)acetic acid

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 (12.18 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 (12.18 mM) (饱和度未知) 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), 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 (12.18 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 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 2% DMSO +30%PEG 300 +5% Tween 80 +ddH2O: 10mg/mL

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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。