cIAP1 (Ki = 1.9 nM); cIAP2 (Ki = 5.1 nM); XIAP (Ki = 66.4 nM)

xevinanapant (AT-406) 盐酸盐有效抑制乳腺癌 MDA-MB-231 和卵巢癌 SK-OV-3 细胞系的细胞生长，IC50 值分别为 144 nM 和 142 nM。 xevinanapant 盐酸盐（0-3 μM；0-48 小时）能够以时间和剂量依赖性方式有效诱导细胞死亡 [1]。

Xevinanapant (AT-406) 盐酸盐可高效抑制 MDA-MB-231 异种移植模型中的肿瘤生长，且对动物的毒性最小[1]。在小鼠、大鼠、非人灵长类动物和犬科动物中研究了 Xevinanapant 盐酸盐的药代动力学 (PK) 参数 [1]。

FL-AT-406（荧光标记的 AT-406）用于开发一套新的 FP 测定法，用于测定 Smac 模拟物与 XIAP、cIAP-1 和 cIAP-2 BIR3 蛋白的结合亲和力。使用固定浓度的 FL-AT-406 和不同浓度的蛋白质直至完全饱和的滴定实验用于计算 FL-AT-406 对每种 IAP 蛋白质的 Kd 值。 Microflu 2 96 孔黑色圆底板用于使用 Infinite M-1000 读板器测量荧光偏振值。对于 XIAP BIR3、cIAP-1 BIR3 和 cIAP-2 BIR3 的实验，将 FL-AT-406（每孔分别为 2、1 和 1 nM）和各种蛋白质浓度添加到终体积为 125 μL 的溶液中。测定缓冲液（100 mM 磷酸钾，pH 7.5，100 g/mL 牛球蛋白，0.02% 叠氮化钠，含 4% DMSO）。彻底混合后，将板在室温下轻轻摇动两到三个小时。在 485 nm 的激发波长和 530 nm 的发射波长下，测量以毫偏振单位 (mP) 为单位的偏振值。然后，使用 Graphpad Prism 5.0 软件，通过拟合 S 形剂量依赖性 FP 增加作为蛋白质浓度的函数来计算平衡解离常数 (Kd)。在 XIAP3 BIR3 的竞争性结合测试中，AT-406 与 20 nM XIAP BIR3 蛋白和 2 nM FL-AT-406 在测定缓冲液（100 mM 磷酸钾，pH 7.5；100 μg/mL 牛 γ-球蛋白；0.02 ％叠氮化钠）。实验中使用 3 nM 蛋白质和 1 nM FL-AT-406 来确定 cIAP1 BIR3 蛋白质的竞争性结合。 5 nM 蛋白质和 1 nM FL-AT-406 用于 cIAP2 BIR3 的竞争性结合测试。使用 Infinite M-1000 读板器，在孵育两到三个小时后确定每个竞争性结合实验的偏振值。使用非线性最小二乘分析，从图中提取 IC50 值或 50% 结合示踪剂被取代时的抑制剂浓度。 PRISM 程序用于拟合曲线。[1]

在这项研究中，研究人员评估了AT-406，一种新的多种IAP蛋白的口服活性拮抗剂，作为单一药物在卵巢癌症细胞中以及与卡铂联合使用的治疗效果和作用机制。他们证明AT-406在60%的体外检测的人类卵巢癌症细胞系中具有显著的单剂活性，并在体内抑制卵巢癌症的进展，并且五分之三的卡铂耐药细胞系对AT-406敏感，突出了AT-406对固有或获得性铂耐药性患者的治疗潜力。此外，我们的体内研究表明，AT-406增强了卡铂诱导的卵巢癌症细胞死亡，表明AT-406使这些细胞对卡铂的反应敏感。从机制上讲，我们证明AT-406诱导的细胞凋亡与其下调XIAP的能力有关，而AT-406在AT-406敏感和抗性细胞系中诱导cIAP1降解。总之，这些结果首次证明了AT-406作为单一药物和与卡铂联合使用的抗卵巢癌症功效，表明AT-406有潜力作为癌症患者的新疗法，尤其是对基于铂的疗法表现出耐药性的患者。[2]

Animal/Disease Models: SCID (severe combined immunodeficient) mouse bearing MDA-MB-231 xenograft tumors [1]
Doses: 30 and 100 mg/kg
Route of Administration: Oral; 5 days per week for 2 weeks
Experimental Results: 30 and 100 mg/kg Strong Inhibits tumor growth, completely inhibiting tumor growth during treatment at 100 mg/kg.

---

In vivo efficacy studies of AT-406[2]
AT-406 was dissolved in DMSO as a stock solution at 200mg/ml. The stock solution was diluted in the vehicle solution, which consists of 10mg/ml hypromellose and 1μl of Tween 80 in PBS, to achieve the final concentration of 10 mg/ml. 5 × 106 OVCAR-3ip cells were injected intraperitoneally (i.p.) into each immunocompromised B6.129S7-Rag1tmMom mouse. Seven days after tumor implantation, the tumor bearing mice were randomly divided into the following treatment groups: control (n = 9), carboplatin (n = 7), AT-406 (n = 5), and combination of AT-406 and carboplatin (n = 7). These groups of mice were treated as follows: each control mouse received 0.2ml of vehicle solution alone by oral gavage every day for 10 d, followed by a 3 d break, and 6 subsequent oral gavage treatments for a total of 16 treatments. AT-406 treatment mice received AT-406 (100 mg/kg) by oral gavage as described for the control treatment mice and the carboplatin treatment mice received carboplatin (40 mg/kg) through intraperitoneal injection twice weekly for two cycles. The combination group received AT-406 and carboplatin treatments simultaneously adhering to the protocols described above for the individual treatment groups. Following an approved IACUC protocol, mouse survival analysis was performed and mice were sacrificed when they appeared moribund or displayed signs of distress, at which time, the mice were considered as dead. At the conclusion of the experiment, mouse tumors and vital organs were removed, fixed, and sectioned for further analyses.[2]

[1]. A potent and orally active antagonist (SM-406/AT-406) of multiple inhibitor of apoptosis proteins (IAPs) in clinical development for cancer treatment. J Med Chem. 2011;54(8):2714-2726.

[2]. AT-406, an orally active antagonist of multiple inhibitor of apoptosis proteins, inhibits progression of human ovarian cancer. Cancer Biol Ther. 2012;13(9):804-811.

Xevinapant is an orally available mimetic of the natural second mitochondrial-derived activator of caspases (Smac) and inhibitor of Inhibitor of Apoptosis Proteins (IAPs), with potential immunomodulating, apoptotic-inducing, chemo-radio-sensitizing and antineoplastic activities. Upon oral administration,xevinapant targets and binds to the Smac binding groove on IAPs, including the direct caspase inhibitor X chromosome-linked IAP (XIAP), and the cellular IAPs 1 (c-IAP1) and 2 (c-IAP2). This inhibits the activities of these IAPs and promotes the induction of apoptosis. Additionally, as xevinapant inhibits the activity of IAPs, it may work synergistically with cytotoxic drugs and/or radiation to overcome tumor cell resistance to apoptosis. As IAPs regulate nuclear factor-kappa B (NFkB) signaling pathways, which drives the expression of genes involved in immune and inflammatory responses, xevinapant may enhance anti-tumor immune responses when administered with certain immunomodulating agents, such as immune checkpoint inhibitors. IAPs are overexpressed by many cancer cell types and suppress both intrinsic and extrinsic apoptosis by binding to and inhibiting active caspases via their baculoviral lAP repeat (BIR) domains. They contribute to chemo-radio-resistance of cancer cells to certain cytotoxic agents and radiation, promote tumor cell survival and are associated with poor prognosis in certain types of cancer. SMAC, a pro-apoptotic mitochondrial protein, is an endogenous inhibitor of the IAPs family of cellular proteins.
See also: Xevinapant Hydrochloride (is active moiety of).

---

Drug Indication
Treatment of head and neck epithelial malignant neoplasms

C32H44CLN5O4

597.31Elemental Analysis

597.308

C, 64.25; H, 7.41; Cl, 5.93; N, 11.71; O, 10.70

1071992-57-8

Xevinapant;1071992-99-8

25022340

White to off-white solid powder

4.473

110.85

3

5

9

41

896

4

C[C@@H](C(=O)N[C@H]1CN(CC[C@H]2CC[C@H](N2C1=O)C(=O)NC(C3=CC=CC=C3)C4=CC=CC=C4)C(=O)CC(C)C)NC

DBXTZCYPHKJCHF-ZZPLZQMBSA-N

InChI=1S/C32H43N5O4.ClH/c1-21(2)19-28(38)36-18-17-25-15-16-27(37(25)32(41)26(20-36)34-30(39)22(3)33-4)31(40)35-29(23-11-7-5-8-12-23)24-13-9-6-10-14-24/h5-14,21-22,25-27,29,33H,15-20H2,1-4H3,(H,34,39)(H,35,40)1H/t22-,25+,26-,27-/m0./s1

(5S,8S,10aR)-N-benzhydryl-5-((S)-2-(methylamino)propanamido)-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocine-8-carboxamide hydrochloride

AT-406; AT 406; AT406; D 1143; Debio 1143; 73T1W2MF9C; UNII-73T1W2MF9C; Xevinapant Hydrochloride; AT-406 HCl; Xevinapant (hydrochloride); (5S,8S,10aR)-N-benzhydryl-5-((S)-2-(methylamino)propanamido)-3-(3-methylbutanoyl)-6-oxodecahydropyrrolo[1,2-a][1,5]diazocine-8-carboxamide hydrochloride; 1071992-57-8 (HCl);N65WC8PXDD; SM 406.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中（例如氮气保护），避免吸湿/受潮和光照。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~175 mg/mL (~292.55 mM)

配方 1 中的溶解度: ≥ 8.75 mg/mL (14.63 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 87.5 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。

---

配方 2 中的溶解度: ≥ 8.75 mg/mL (14.63 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 87.5 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。

---

View More

配方 3 中的溶解度: ≥ 8.75 mg/mL (14.63 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 87.5 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，混合均匀。

---

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。