TVB-3664

在 CaCo2、HT29 和 LIM2405 细胞系中，TVB-3664（0-1 μM，7 天）表现出抗肿瘤作用 [1]。 TVB-3664 会降低来自血液和实体恶性肿瘤的几种肿瘤细胞系的活力[1]。

TVB-3664（Pt 2614 和 Pt 2449PT 为 3 mg/kg）或 Pt 2402 和 Pt 2449LM 为 6 mg/kg）口服管饲治疗 4 周后，肿瘤体积和重量显着下降。对于 Pt 2614 PDX 模型，平均肿瘤重量分别减少了 30%、37.5% 和 51.5% [1]。

TMA分析[1]
在匹配的正常结肠粘膜和肿瘤组织中分析FASN表达的免疫反应性评分，这些组织来自在UK Chandler Medical Center接受手术的诊断为I-IV期CRC的患者（TMA ID BH15991A，n=57个正常组织和56个肿瘤组织）。病理学家盲目地进行评分。免疫反应性评分通过染色强度值（0，无染色；1，弱染色；2，中等染色；3，强染色）和阳性肿瘤细胞百分比值（0：无阳性细胞；1，0-10%；2，11-50%；3，51-100%阳性）的乘积来确定

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核磁共振分析[1]
NMR光谱在配备有3毫米反向三共振HCN冷探针的安捷伦DD2 14.1T光谱仪上获得。在288k下记录1D质子光谱，采集时间为2s，弛豫延迟为4s，在此期间照射残余HOD共振。记录1D 1H[13C]HSQC光谱，采集时间为0.25s，弛豫延迟为1.75s。 内部DSS-d6用作化学位移参考并用于绝对定量。MNOVA用于NMR光谱处理。在分阶段和基线校正后，使用MNOVA中的峰值拟合程序对代谢物进行量化，并使用内部数据库进行分配。同位素被量化为绝对富集和部分富集，如前所述。通过BCA分析将分配的代谢物标准化为mg蛋白质。

细胞增殖测定[1]
细胞类型： CaCo2、HT29 和 LIM2405 细胞系。
测试浓度：0-1 μM。
孵化持续时间：7天。
实验结果：证明具有抗肿瘤活性。

Animal/Disease Models: Colorectal cancer (CRC) PDX models in NOD-SCID-IL2rg-/- (NSG) mice using specimens collected from patients who had undergone surgery for resection of primary CRC or CRC metastasis[1].
Doses: 3 mg/kg (Pt 2614 and Pt 2449PT) or 6 mg/kg (Pt 2402 and Pt 2449LM).
Route of Administration: po (oral gavage) daily for 4 weeks.
Experimental Results: Led to a significant reduction in tumor volume and tumor weight in Pt 2614, Pt 2449PT, and Pt 2402 PDX models, with an average reduction in tumor weight of 30%, 37.5% and 51.5%, respectively.

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Tissues were obtained from consented patients with Stage II-IV CRC who had undergone surgery at UK Medical Center. 6–8-week-old NSG mice (NOD.Cg-Prkdc Il2rg /SzJ) from The Jackson Laboratory were used for PDX models. All procedures were performed using protocols approved by the UK Animal Care and Use Committee. Briefly, CRC tissues (2–5 mm) obtained from CRC patients of both sexes were implanted subcutaneously into their flanks in a small pocket surgically created under the skin. Established tumors were designated as generation 0 (G0). Tumor tissues from G0 were minced and mixed with Matrigel to ensure homogeneous distribution of tissues among mice and allow implantation of an equal volume of tumor tissues into the flank. To preserve histopathological and genomic characteristics of clinical CRC tumors and recapitulate the differential responses of CRC tumors to FASN inhibitors, all established PDX models were treated at G1 with exception of the Pt 2387 case, which was treated at G2. When tumors reached 100 mm3, the mice were randomized according to animal weight and tumor size. For evaluation as monotherapy, treatment (n = 5) and control (n = 5) groups were given TVB-3664 (3–6 µg/kg) vs vehicle (30% PEG400) by oral gavage daily for 4–6 weeks. Tumors were measured once per week by a digital caliper and tumor volume was calculated using the formula: TV = width2 × length × 0.52 as previously described. Tumor weight was measured at the end of the experiment. Blood samples were collected through cardiac puncture at the time of sacrifice. Mice were fasted and then injected i.p. with 2 g kg–1 body mass of 13C6-glucose 16 h prior to sacrifice as previously described, to allow SIRM tracing of cellular metabolites of tumors. 1 h after injection, blood samples were collected and separated into plasma and blood cells by centrifugation (4° C, 3,500 × g, 15 min). Mice were euthanized and tumor tissue was collected for IHC and flash frozen immediately in liquid N2 for protein and metabolic analysis.[1]

[1]. Preclinical evaluation of novel fatty acid synthase inhibitors in primary colorectal cancer cells and a patient-derived xenograft model of colorectal cancer. Oncotarget. 2018 May 15;9(37):24787-24800.

[2]. FASN Inhibition and Taxane Treatment Combine to Enhance Anti-tumor Efficacy in Diverse Xenograft Tumor Models through Disruption of Tubulin Palmitoylation and Microtubule Organization and FASN Inhibition-Mediated Effects on Oncogenic Signaling and Gene Expression. EBioMedicine. 2017 Feb;16:51-62.

Fatty Acid Synthase (FASN), a key enzyme of de novo lipogenesis, is upregulated in many cancers including colorectal cancer (CRC); increased FASN expression is associated with poor prognosis. Potent FASN inhibitors (TVBs) developed by 3-V Biosciences demonstrate anti-tumor activity in vitro and in vivo and a favorable tolerability profile in a Phase I clinical trial. However, CRC characteristics associated with responsiveness to FASN inhibition are not fully understood. We evaluated the effect of TVB-3664 on tumor growth in nine CRC patient-derived xenografts (PDXs) and investigated molecular and metabolic changes associated with CRC responsiveness to FASN inhibition. CRC cells and PDXs showed a wide range of sensitivity to FASN inhibition. TVB-3664 treatment showed significant response (reduced tumor volume) in 30% of cases. Anti-tumor effect of TVB-3664 was associated with a significant decrease in a pool of adenine nucleotides and alterations in lipid composition including a significant reduction in fatty acids and phospholipids and an increase in lactosylceramide and sphingomyelin in PDXs sensitive to FASN inhibition. Moreover, Akt, Erk1/2 and AMPK were major oncogenic pathways altered by TVBs. In summary, we demonstrated that novel TVB inhibitors show anti-tumor activity in CRC and this activity is associated with a decrease in activation of Akt and Erk1/2 oncogenic pathways and significant alteration of lipid composition of tumors. Further understanding of genetic and metabolic characteristics of tumors susceptible to FASN inhibition may enable patient selection and personalized medicine approaches in CRC.
References: https://pubmed.ncbi.nlm.nih.gov/29872506/

C25H23F3N4O2

468.48

468.177

C, 64.10; H, 4.95; F, 12.17; N, 11.96; O, 6.83

2097262-58-1

129101638

White to off-white solid powder

1.36±0.1 g/cm3(Predicted)

665.7±55.0 °C(Predicted)

3.8

82Ų

1

7

5

34

770

0

YFEOVRCUSPPGFZ-UHFFFAOYSA-N

InChI=1S/C25H23F3N4O2/c1-14-8-15(2)20(9-19(14)22-21(13-34-3)30-24(31-22)25(26,27)28)23(33)32-11-18(12-32)17-6-4-16(10-29)5-7-17/h4-9,18H,11-13H2,1-3H3,(H,30,31)

4-(1-(5-(4-(methoxymethyl)-2-(trifluoromethyl)-1H-imidazol-5-yl)-2,4-dimethylbenzoyl)azetidin-3-yl)benzonitrile

TVB3664; TVB 3664; TVB-3664

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)

DMSO : ~10 mg/mL (~21.35 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.34 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 2 中的溶解度: ≥ 2.08 mg/mL (4.44 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 20.8 mg/mL澄清的DMSO储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入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 3 中的溶解度: ≥ 2.08 mg/mL (4.44 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 20.8 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。