CBR-5884

Phosphoglycerate dehydrogenase/PHGDH

CBR-5884（15 或 30 μM；3-5 天）专门防止具有强烈丝氨酸合成倾向的乳腺癌和黑色素瘤细胞系增殖。

体内研究显示，CBR-5884显著延缓肿瘤生长，组织学分析表明CBR-584的安全性。最后，利用患者来源的类器官（PDO）模型来探索CBR-5884对EOC细胞的临床前疗效，结果表明，CBR-584在EOC PDO模型中阻碍增殖并下调ITGB4的表达。[2]

PHGDH小分子抑制剂的筛选。[1]
一种适用于高通量筛选（HTS）的PHGDH活性的体外酶法测定是通过将3-PG氧化后NADH的产生与使用黄递酶作为偶联酶将雷沙祖林还原为间苯二酚偶联而开发的。因此，间苯二酚荧光作为PHGDH活性的替代物。该测定被小型化为1536孔格式，Z因子>0.75，表明该测定具有高质量。在13μM下以单点形式筛选了80万个小分子的文库。将阈值Z分数设置为−3，对应于至少50%的PHGDH抑制，可获得0.5%的命中率，产生3906次命中。假定的命中率以一式三份的形式重新确认，并对黄递酶进行反筛选，以排除针对黄递酶的假阳性。计数器筛选消除了3498种化合物，得到408种PHGDH抑制剂。
设计了一种基于命中效力和选择性的试验策略。我们推断，与打击多种脱氢酶的化合物相比，PHGDH特异性抑制剂将最大限度地降低一般细胞毒性。因此，测定了一组NAD（P）+依赖性脱氢酶的半数最大抑制浓度（IC50），这些脱氢酶包括PHGDH、异柠檬酸脱氢酶（IDH1）、苹果酸脱氢酶（MDH1）和3α-羟基类固醇脱氢酶（3α-HSD）。化合物对PHGDH的选择性提高了至少四倍以供进一步分析。基于这一试验，选择了七种最有效的PHGDH抑制剂作为先导化合物，用于在基于细胞的测定中进行评估；选定的结构如图1E所示。许多这些化合物可能靶向巯基，因此可能与PHGDH半胱氨酸残基反应。例如，CBR-587和CBR-6936都含有巯基反应性二硫中心。有趣的是，CBR-5807（二硫仑）是一种已批准用于治疗酒精中毒的药物，已知通过与巯基反应抑制乙醛脱氢酶。

细胞增殖测定[1]
细胞类型：乳腺癌和黑色素瘤细胞系
测试浓度：15 或 30 μM
孵育时间：3-5天
实验结果：抑制黑色素瘤和乳腺癌细胞系的增殖。

The study utilized 12 female BALB/c nude mice aged 4–6 weeks, procured from the Experimental Animal Center of Southern Medical University. These mice were housed in autoclaved, ventilated cages and provided with autoclaved water. All mice used were bred in the specific pathogen free laboratory. The mice received a subcutaneous injection of 1 × 106 ID8 cells in the right armpit. When the tumors reached an average volume of 65 mm3, the mice were randomly divided into two groups using the random number method: (1) an experimental group, wherein the mice received intragastric administration of CBR-5884 (70 mg/kg, qd; n = 6); and (2) a control group, wherein the mice received intragastric administration of an equal volume of vehicle (corn oil) (n = 6). CBR-5884 treatment lasted for 12 consecutive days, with continuous drug administration. Tumors were resected when the average tumor volume in the control group reached 400 mm3. Tumor growth was tracked using caliper measurements, and tumor volume was calculated using the formula: length × width2/2. Subsequently, images of mice and tumors were captured, and tumors were weighed. Liver, spleen, kidney, and tumors were subjected to histological analysis after hematoxylin–eosin (H&E) staining. The maximal tumor size/burden permitted by our institutional review board is 10% of body weight and mean tumor diameter = or > 15 mm in adult mice (~ 25 g). The maximal tumor size/burden permitted by our institutional review board was not exceeded.[2]

[1]. Identification of a small molecule inhibitor of 3-phosphoglycerate dehydrogenase to target serine biosynthesis in cancers Proc Natl Acad Sci U S A, 2016 Feb 16, 113(7):1778-83.

Reprogramming of the serine synthesis pathway (SSP) is intricately linked to the progression of epithelial ovarian cancer (EOC). CBR-5884, a selective small-molecule inhibitor targeting phosphoglycerate dehydrogenase (PHGDH), effectively impedes the de novo synthesis of serine within cancer cells. This study aimed to evaluate the inhibitory effect of CBR-5884 on EOC cells and delineate its specific mechanism, thereby proposing a novel therapeutic approach for treating EOC. The suppression of serine biosynthesis after CBR-5884 treatment was evaluated using RNA sequencing and a serine assay kit, and the results showed that CBR-5884 effectively downregulated serine biosynthesis in EOC cells, particularly those expressing high levels of PHGDH. In vitro studies revealed that CBR-5884 demonstrated significant antitumor effects and suppressed migration and invasion of EOC cells through down-regulation of the integrin subunit beta 4 (ITGB4)/extracellular signal-regulated kinase (ERK)/epithelial–mesenchymal transition signal axis. Additionally, CBR-5884 mitigated the stemness of EOC cells and heightened their sensitivity to chemotherapy. Moreover, in vivo studies revealed that CBR-5884 significantly delayed tumor growth, with histological analysis indicating the safety profile of CBR-5884. Finally, the patient-derived organoid (PDO) models were utilized to explore the preclinical efficacy of CBR-5884 against EOC cells, and the results unveiled that CBR-5884 impeded proliferation and downregulated the expression of ITGB4 in EOC PDO models. Our findings supports the anticancer properties of CBR-5884 in EOC cells exhibiting high PHGDH expression, manifesting through the suppression of proliferation, migration, and invasion, while enhancing chemotherapy sensitivity, suggesting that CBR-5884 holds promise as an efficacious strategy for the treatment of EOC[2].

C14H12N2O4S2

336.386

336.023

C, 49.99; H, 3.60; N, 8.33; O, 19.02; S, 19.06

681159-27-3

4674993

Typically exists as light yellow to yellow solids at room temperature

1.4±0.1 g/cm3

378.1±42.0 °C at 760 mmHg

182.5±27.9 °C

0.0±0.9 mmHg at 25°C

1.628

4.15

146Ų

1

7

6

22

479

0

O=C(C1=C(C)C(SC#N)=C(NC(C2=CC=CO2)=O)S1)OCC

QBVIRPJBDIZKBC-UHFFFAOYSA-N

InChI=1S/C14H12N2O4S2/c1-3-19-14(18)11-8(2)10(21-7-15)13(22-11)16-12(17)9-5-4-6-20-9/h4-6H,3H2,1-2H3,(H,16,17)

ethyl 5-(furan-2-carboxamido)-3-methyl-4-thiocyanatothiophene-2-carboxylate

CBR-5884; CBR 5884; CBR5884.

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 : ~50 mg/mL (~148.64 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (6.18 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 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;
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7、 以上所有助溶剂都可在 Invivochem.cn网站购买。