Tazemetostat de(methyl morpholine)-COOH

Intermediate for PROTAC EZH2 degrader

传统的EZH2抑制剂是为了抑制酶甲基化活性而开发的，由于EZH2在癌症发展中的非酶功能，它们可能具有治疗局限性。在这里，我们报道了基于蛋白水解靶向嵌合体（PROTAC）的EZH2降解物靶向淋巴瘤中的整个EZH2。设计并合成了两个系列的EZH2降解剂，以劫持含有von Hippel-Lindau （VHL）或cereblon （CRBN）的E3连接酶系统，并且一些基于VHL的化合物能够介导EZH2的降解。两种最佳降降剂YM181和YM281在弥漫性大b细胞淋巴瘤（DLBCL）和其他亚型淋巴瘤中诱导了强大的细胞活力抑制，优于临床使用的EZH2抑制剂EPZ6438（他泽美他），后者仅对DLBCL有效。EZH2降解物在淋巴瘤异种移植物和患者源性原发性淋巴瘤细胞中显示出良好的抗肿瘤活性。我们的研究表明EZH2降解物比EZH2抑制剂具有更好的治疗活性，这可能为治疗淋巴瘤提供潜在的抗癌策略。

[1]. Design, Synthesis, and Evaluation of VHL-Based EZH2 Degraders to Enhance Therapeutic Activity against Lymphoma. J Med Chem. 2021 Jul 22;64(14):10167-10184.

In this study, the therapeutic effect of EZH2 inhibitors was limited to the diffuse large B-cell lymphoma (DLBCL) cell line tested (Figure 1A). We wanted to explore whether a method for directly degrading EZH2 using PROTAC technology could be developed to enhance its targeting ability against other types of lymphoma cells. In this study, we developed PROTAC-based EZH2 degraders and investigated their EZH2 degradation efficiency and therapeutic effects against different types of lymphoma in vitro and in vivo. Our study showed that only VHL-targeting compounds could degrade EZH2 through suitable linker groups of 7 or 9 atoms in length. Compared to the parent EZH2 inhibitor EPZ6438, our two optimal EZH2 degraders, YM181 and YM281, showed higher selectivity for EZH2 than EZH1 and exhibited potent antiproliferative activity in both DLBCL and other types of lymphoma cell lines. Furthermore, at effective doses, the EZH2 degraders significantly inhibited the in vivo growth of lymphoma xenografts without significant toxicity. However, YM281 and YM181 exhibit incomplete EZH2 degradation at low concentrations and limited efficacy in inhibiting cell viability, indicating room for further optimization. Future research may necessitate structure-activity relationship studies on different linker backbones with similar linker lengths to YM181 and YM281 to obtain more effective EZH2 degraders. Furthermore, some EZH2-independent tumorigenesis cancer cells, such as pancreatic cancer cells AsPC1 and lung cancer cells NCI-H460, are insensitive to YM181 and YM281, despite these compounds reducing their EZH2 levels (Figure S6). Cell permeability measurements using Caco-2 cells showed that the apparent permeability of both YM181 and YM281 was significantly lower than that of their parent EZH2 inhibitor EPZ6438 (Table S2), suggesting the need for further structural optimization to improve their oral bioavailability. Overall, our results suggest that EZH2 degraders may have better therapeutic potential than EZH2 inhibitors in treating lymphoma. We are investigating the exact mechanism of action of EZH2 degraders and their application in other cancers. [1]

C30H35N3O5

517.616008043289

517.257

2685873-44-1

165437231

Off-white to yellow solid powder

4.2

108

3

6

8

38

946

0

MIEXIPQREXYLRA-UHFFFAOYSA-N

InChI=1S/C30H35N3O5/c1-5-33(24-10-12-38-13-11-24)27-16-23(21-6-8-22(9-7-21)30(36)37)15-25(20(27)4)28(34)31-17-26-18(2)14-19(3)32-29(26)35/h6-9,14-16,24H,5,10-13,17H2,1-4H3,(H,31,34)(H,32,35)(H,36,37)

4-[3-[(4,6-dimethyl-2-oxo-1H-pyridin-3-yl)methylcarbamoyl]-5-[ethyl(oxan-4-yl)amino]-4-methylphenyl]benzoic acid

Tazemetostat de(methyl morpholine)-COOH; 2685873-44-1; CHEMBL5398431; BDBM50633529;

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 :~220 mg/mL (~425.02 mM)

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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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。