DNA-PK (IC50 <3 nM)

M3814在体外使多种肿瘤细胞系对放射治疗敏感[2]

---

M3814通过抑制关键的DSB DNA损伤修复机制来靶向肿瘤细胞的生长和存活。M3814的抗肿瘤作用取决于癌症细胞中DNA修复和检查点信号的功能，这些细胞应对DSB的能力降低，导致细胞死亡。因此，DNA-PK抑制的基本原理是增加IR产生的DSB DNA损伤量。一系列非临床实验的目的是证明IR与M3814联合使用比单独使用IR更有效[1]。

与IR联合使用，M3814在所有6种人类癌症小鼠模型中均显示出疗效，IR的作用增强证明了这一点。在所有模型中，与单独使用IR相比，每天2 Gy的剂量与M3814联合使用1周可诱导静态显著的肿瘤生长抑制。在两种模型中，FaDu和NCIH460、M3814诱导了肿瘤消退。在FaDu模型中，在25和50mg/Kg剂量下，IR联合组（每部分2 Gy，6周，每周5天：总剂量60 Gy）没有观察到肿瘤再生（实验持续时间>100天）。在仅IR组中，没有观察到任何肿瘤反应。这些效应可能是抑制DNA-PK活性的结果，如测量FaDu肿瘤组织中DNA-PK的自磷酸化所示。在任何研究中，M3814单独或与IR联合使用均未导致小鼠体重明显减轻或出现视觉毒性迹象。[1]

---

M3814显著增强了电离辐射在体内的抗肿瘤活性，并采用临床相关的分割辐射方案实现了肿瘤的完全消退。这些效应是由于抑制了DNA-PK蛋白激酶活性，如人类肿瘤细胞系中DNA-PK自磷酸化水平所示，异种移植物肿瘤M3814目前正在PhI临床试验中进行研究[2]。

The efficacy of M3814 in combination with IR was evaluated in 6 human xenograft models (HCT116, FaDu, NCI-H460, A549, Capan-1, BxPC3) in mice representing 4 different cancer types (colon, head and neck, lung, and pancreas). Tumor cells were injected s.c. into nude mice, and treatment started when palpable tumors were established (w100-200 mm3 ). M3814 was given orally at different doses (25-300 mg/kg) 10 min prior to IR. IR was applied using a radiation therapy device for small rodents calibrated to deliver 2 Gy. Autophosphorylation of DNA-PK (serine2056 ) in FaDu tumor lysates was measured by immunoassay to assess pharmacological inhibition by M3814.[1]

[1]. M3814, a DNA-dependent Protein Kinase Inhibitor"n(DNA-PKi), Potentiates the Effect of Ionizing Radiation (IR) in"nXenotransplanted Tumors in Nude Mice. IJROBP. 2016; 94, 940-941.

[2]. Abstract 1658: M3814, a novel investigational DNA-PK"ninhibitor: enhancing the effect of fractionated radiotherapy leading to"ncomplete regression of tumors in mice. AACR; Cancer Res 2016;76(14"nSuppl):Abstract nr 1658.

[3]. Arylchinazoline. WO2014183850A1.

Nedisertib is under investigation in clinical trial NCT03770689 (Study of M3814 in Combination With Capecitabine and Radiotherapy in Rectal Cancer).
Peposertib is an orally bioavailable inhibitor of DNA-dependent protein kinase (DNA-PK) with potential antineoplastic activity, and potential sensitizing and enhancing activities for both chemo- and radiotherapies. Upon administration, peposertib binds to and inhibits the activity of DNA-PK, thereby interfering with the non-homologous end joining (NHEJ) process and preventing repair of DNA double strand breaks (DSBs) caused by ionizing radiation or chemotherapeutic treatment. This increases chemo- and radiotherapy cytotoxicity and leads to enhanced tumor cell death. The enhanced ability of tumor cells to repair DSBs plays a major role in the resistance of tumor cells to chemo- and radiotherapy; DNA-PK plays a key role in the NHEJ pathway and DSB repair.

---

M3814 is active in nonclinical experiments in combination with IR. Strong antitumor activity was observed in several xenograft models with complete regressions of tumors upon application of the established clinical IR schedule of 2-Gy fractions for 6 weeks in the FaDu model (squamous cell carcinomas of the head and neck). Clinical evaluation of M3814 is ongoing.[1]

---

Physical or chemical agents that damage DNA such as ionizing radiation are among the most widely used classes of cancer therapeutics today. Double strand breaks (DSB) generated in DNA by radiation induce multitude of cellular responses, including DNA repair, cell cycle arrest or cell death if the damage is left unrepaired. A complex set of molecular events are responsible for DNA repair via two major mechanisms - homologous recombination (HR) or non-homologous end joining (NHEJ). DNA-PKcs with its regulatory protein subunits, Ku70 and Ku80, is an integral component of NHEJ and considered an attractive intervention point to inhibit DNA repair. We have developed an orally bioavailable, highly potent, and selective inhibitor of DNA-PK, M3814, for cancer therapy in combination with DNA damaging modalities such as radiation, and radio-chemotherapy. Here, we present the preclinical characterization of M3814 using biochemical, cellular and human tumor xenograft models. [2]

C24H21CLFN5O3

481.9066

481.131

C, 59.82; H, 4.39; Cl, 7.36; F, 3.94; N, 14.53; O, 9.96

1637542-33-6

1637542-33-6 (S-isomer); 1637542-34-7 (racemate); 1637542-32-5 (R-isomer)

86292849

Light yellow to yellow solid powder

2.8

93.5

1

9

5

34

662

1

ClC1C=C(C(=CC=1[C@@H](C1C=CC(=NN=1)OC)O)C1C2C=CC(=CC=2N=CN=1)N1CCOCC1)F

MOWXJLUYGFNTAL-DEOSSOPVSA-N

InChI=1S/C24H21ClFN5O3/c1-33-22-5-4-20(29-30-22)24(32)16-11-17(19(26)12-18(16)25)23-15-3-2-14(10-21(15)27-13-28-23)31-6-8-34-9-7-31/h2-5,10-13,24,32H,6-9H2,1H3/t24-/m0/s1

(S)-(2-chloro-4-fluoro-5-(7-morpholinoquinazolin-4-yl)phenyl)(6-methoxypyridazin-3-yl)methanol

MSC2490484A; MSC-2490484A; nedisertib; peposertib; 1637542-33-6; M3814; M-3814; MSC-2490484-A; Nedisertib [INN]; M-3814(nedisertib); MSC 2490484A; M3814; M-3814; M 3814

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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。

---

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。

View More

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)

---

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。

View More

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

---

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。

---

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。