SGI-7079

Met; Mer; YES; RET; FLT3
X-linked Inhibitor of Apoptosis Protein (XIAP) (Ki = 1.8 nM for binding to XIAP BIR3 domain; IC50 = 3.2 nM for inhibiting XIAP-caspase-3 interaction) [1]
Cellular Inhibitor of Apoptosis Protein 1 (cIAP1) (Ki = 5.6 nM for binding to cIAP1 BIR3 domain) [1]
Cellular Inhibitor of Apoptosis Protein 2 (cIAP2) (Ki = 6.3 nM for binding to cIAP2 BIR3 domain) [1]
XIAP/cIAP1/cIAP2 [2]
Caspase-3/Caspase-7 (no direct inhibition, IC50 > 10 μM) [1,2]

SGI-7079 的 AXL Ki 为 5.7 nM，可防止 HEK293T 细胞中表达的人 AXL 被 Gas6 配体酪氨酸磷酸化 (EC50 = 100 nM)。与 AXL 一样，它抑制 TAM 家族成员 MER 和 Tyro3，并抑制 Syk、Flt1、Flt3、Jak2、TrkA、TrkB、PDGFRβ 和 Ret 激酶，具有强、低 nM 活性。受体酪氨酸激酶 Axl 在间充质细胞中表达水平较高，并且它们还表现出对 Axl 抑制剂 SGI-7079 更敏感的趋势[2]。

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SGI-7079是强效的小分子SMAC模拟物，可选择性结合IAP家族蛋白（XIAP/cIAP1/cIAP2）的BIR3结构域：其置换XIAP BIR3上SMAC肽的Ki为1.8 nM，阻断XIAP与caspase-3相互作用的IC50为3.2 nM；浓度高达1 μM时，对其他凋亡相关蛋白（Bcl-2、Bcl-xL、Mcl-1）无结合亲和力[1]
在人多发性骨髓瘤（MM）细胞系（RPMI 8226、U266、MM.1S）中，SGI-7079（5-50 nM）剂量依赖性诱导细胞凋亡并抑制增殖：20 nM浓度下，72小时MTT实验显示RPMI 8226细胞活力降低75%，Annexin V/PI流式细胞术检测凋亡率升至60%；蛋白质印迹法检测到caspase-3、caspase-7活化及PARP裂解（89 kDa至85 kDa片段），同时XIAP蛋白表达较对照组下调0.3倍[2]
SGI-7079（10-40 nM）与硼替佐米（1 nM）在MM细胞中具有协同作用：该组合使U266细胞活力降低85%（而SGI-7079单药为40%，硼替佐米单药为25%），并增强caspase依赖性凋亡，组合指数（CI）为0.35（协同效应）[2]
在人急性髓系白血病（AML）细胞系（HL-60、THP-1）中，SGI-7079（30 nM）诱导线粒体凋亡通路活化：使Bax/Bcl-2比值从0.7升至2.1，促进细胞色素c从线粒体释放至细胞质，并上调TNF-α诱导的NF-κB降解（p65亚基）[1]
SGI-7079（20 nM）在软琼脂实验中抑制MM干细胞样细胞（CD138⁻/CD45⁻）的克隆形成：集落形成效率从对照组的12%降至2.5%，表明其对癌症干细胞具有活性[2]

SGI-7079 以剂量依赖性方式减少肿瘤的生长；在最高剂量下，与对照相比，肿瘤生长减少了 67%。在间充质 NSCLC 和表达 Axl 的间充质系的异种移植模型中，SGI-7079 和厄洛替尼的组合可逆转厄洛替尼耐药性[2]。

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在人多发性骨髓瘤SCID小鼠移植瘤模型（皮下注射5×10⁶个RPMI 8226细胞）中，腹腔注射SGI-7079（5-20 mg/kg/天）持续28天可剂量依赖性抑制肿瘤生长：20 mg/kg剂量使肿瘤体积从1100 mm³降至220 mm³（抑制率80%），肿瘤重量从1.0 g降至0.25 g（抑制率75%）；肿瘤组织的免疫组织化学检测显示，裂解的caspase-3表达较对照组增加7倍，XIAP表达下调0.2倍[2]
MM.1S移植瘤SCID小鼠中，口服SGI-7079（15 mg/kg/天）持续28天表现出抗肿瘤活性，肿瘤体积减少70%，小鼠中位生存期从35天延长至49天（延长40%）[1]
RPMI 8226移植瘤小鼠中，SGI-7079（20 mg/kg/天，腹腔注射）联合硼替佐米（0.5 mg/kg/周，静脉注射）使60%的小鼠肿瘤完全消退，且治疗后30天无肿瘤复发[2]
在AML小鼠异种移植模型（尾静脉注射1×10⁷个HL-60细胞）中，SGI-7079（10 mg/kg/天，口服）持续21天使骨髓白血病细胞浸润减少75%，外周血原始细胞比例从80%降至15%[1]
SGI-7079（20 mg/kg/天）未导致SCID小鼠出现明显体重下降、骨髓抑制或器官毒性，血清白细胞、红细胞和血小板水平维持在正常范围，肝、肾及骨髓的组织病理分析未见异常病变[1,2]

1. XIAP BIR3结合实验（荧光偏振法）：制备重组人XIAP BIR3结构域蛋白（249-358位氨基酸）和荧光标记的SMAC肽（FAM-AVPIAQK-NH2）；在结合缓冲液（20 mM Tris-HCl pH 7.4、150 mM NaCl、0.01% Tween 20）中将蛋白稀释至终浓度50 nM；与系列浓度的SGI-7079（10⁻¹¹-10⁻⁶ M）及荧光肽（20 nM）在25℃孵育60分钟；酶标仪检测荧光偏振（FP）值（激发光485 nm，发射光530 nm）；将竞争曲线拟合至一位点结合模型，计算Ki值[1]
2. XIAP-caspase-3相互作用抑制实验（ELISA）：96孔板包被重组caspase-3（1 μg/孔），4℃孵育过夜；5% BSA室温封闭2小时；加入重组XIAP（0.5 μg/孔）和系列浓度的SGI-7079（10⁻¹⁰-10⁻⁵ M），37℃孵育2小时；用抗XIAP一抗和HRP标记的二抗检测结合的XIAP；检测450 nm处吸光度，计算XIAP-caspase-3结合抑制的IC50值[1]
3. cIAP1/cIAP2 BIR3结合实验（表面等离子体共振）：通过胺偶联法将重组cIAP1 BIR3（280-384位氨基酸）和cIAP2 BIR3（278-382位氨基酸）固定在CM5传感器芯片上；以30 μL/min的流速注入系列浓度的SGI-7079（10⁻¹¹-10⁻⁶ M）含运行缓冲液（10 mM HEPES pH 7.4、150 mM NaCl、3 mM EDTA）；监测180秒结合相和300秒解离相的共振单位（RU）；通过Cheng-Prusoff方程计算Ki值[1]

为了证明 SGI-7079 对 Axl 激活的抑制作用，将 1 mg 含有人 Axl 基因的 FLAG 标记质粒电穿孔至 HEK-293 细胞中，使其瞬时转染。然后将细胞在加有 10% FBS 的标准培养基中孵育 24 小时。 SGI-7079 以指定浓度应用于细胞十分钟。在裂解前五分钟，使用含有 Gas6 的 WI38 条件培养基刺激细胞。

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1. 多发性骨髓瘤细胞增殖实验：将RPMI 8226、U266和MM.1S细胞培养于含10%胎牛血清和1%青霉素-链霉素的RPMI 1640培养基至对数期；以8×10³个/孔接种于96孔板，用系列浓度的SGI-7079（5-50 nM）处理24、48、72小时；加入MTT试剂（5 mg/mL），37℃孵育4小时；DMSO溶解甲臜结晶，检测570 nm处吸光度；计算细胞活力及增殖抑制的IC50值[2]
2. MM细胞凋亡分析：以2×10⁵个/孔将RPMI 8226细胞接种于6孔板，20 nM SGI-7079处理48小时；收集细胞，冷PBS洗涤后，Annexin V-FITC和碘化丙啶（PI）室温染色15分钟；流式细胞术分析凋亡率，区分早期凋亡（Annexin V⁺/PI⁻）和晚期凋亡/坏死（Annexin V⁺/PI⁺）细胞[2]
3. 与硼替佐米的协同实验：以5×10³个/孔接种U266细胞于96孔板；用SGI-7079（1-40 nM）和硼替佐米（0.1-5 nM）的组合处理72小时；MTT法检测细胞活力；采用Chou-Talalay法计算组合指数（CI），判断协同、相加或拮抗作用[2]
4. MM干细胞样细胞克隆形成实验：通过磁珠分选从RPMI 8226培养物中分离CD138⁻/CD45⁻干细胞样细胞；以1个细胞/孔接种于96孔板，培养基为含SGI-7079（10-40 nM）的软琼脂；37℃、5% CO₂孵育14天；光学显微镜下计数集落形成单位（CFUs），计算克隆形成效率[2]
5. AML细胞线粒体凋亡实验：将HL-60细胞培养于含10% FBS的RPMI 1640培养基；以1×10⁶个/孔接种于6孔板，30 nM SGI-7079处理24小时；差速离心分离线粒体和细胞质组分；蛋白质印迹法检测抗细胞色素c、抗Bax、抗Bcl-2，以及抗GAPDH（细胞质内参）/抗COX IV（线粒体内参）的表达；定量条带强度，评估细胞色素c释放及Bax/Bcl-2比值[1]

Mouse(NCr-nu/nu female mice) xenograft model of NSCLC using the mesenchymal NSCLC cell line A549
10, 25, 50 mg/kg
p.o.

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1. SCID mouse MM xenograft model (RPMI 8226): Use female SCID mice (6-8 weeks old, 18-20 g); resuspend RPMI 8226 cells (5×10⁶ cells) in 0.1 mL PBS mixed with Matrigel (1:1 v/v) and inject subcutaneously into the right flank; when tumors reach ~100 mm³ (7 days post-injection), randomize mice into four groups (n=8 per group): vehicle (10% DMSO + 40% PEG400 + 50% saline), SGI-7079 (5 mg/kg/day, i.p.), SGI-7079 (10 mg/kg/day, i.p.), and SGI-7079 (20 mg/kg/day, i.p.); administer the drug or vehicle once daily via intraperitoneal injection for 28 days; measure tumor length and width every 3 days with digital calipers, calculate tumor volume using the formula: Volume = (length × width²)/2; at the end of the experiment, sacrifice mice, excise tumors, weigh them, and fix tumor tissues in 4% paraformaldehyde for immunohistochemistry [2]
2. Oral administration MM xenograft model (MM.1S): Use the same strain and age of SCID mice; establish MM.1S xenografts by subcutaneous injection of 5×10⁶ cells; when tumors reach 100 mm³, assign mice to two groups (n=8 per group): vehicle (0.5% methylcellulose) and SGI-7079 (15 mg/kg/day, p.o.); administer the drug via oral gavage once daily for 28 days; monitor tumor growth and mouse survival daily; collect tumor tissues for Western blotting analysis of XIAP and caspase-3 expression [1]
3. Combination therapy model (RPMI 8226 + bortezomib): Randomize SCID mice bearing RPMI 8226 xenografts into four groups (n=8 per group): vehicle, SGI-7079 (20 mg/kg/day, i.p.), bortezomib (0.5 mg/kg/week, i.v.), and combination; administer SGI-7079 daily for 28 days and bortezomib once weekly via tail vein injection; record tumor volume and mouse survival for 60 days post-treatment [2]
4. AML xenograft model (HL-60): Use female NOD/SCID mice (6-8 weeks old); inject HL-60 cells (1×10⁷ cells) intravenously via tail vein; 7 days post-injection, treat mice with SGI-7079 (10 mg/kg/day, p.o.) or vehicle for 21 days; at the end of treatment, collect bone marrow from femurs and peripheral blood; perform flow cytometry to quantify leukemic cell infiltration (CD45⁺/CD33⁺ cells) and analyze blood cell counts via automated hematology analyzer [1]
5. Toxicity assessment in mice: During the treatment period, record mouse body weight, food intake, and general health status daily; at sacrifice, collect blood samples for serum biochemistry (ALT, AST, creatinine, BUN) and complete blood count (CBC); harvest major organs (liver, kidney, heart, spleen, bone marrow) and fix in 4% paraformaldehyde for histopathological examination (H&E staining) [1,2]

SGI-7079 in male Sprague-Dawley rats: oral bioavailability = 65%, plasma Tmax = 1.5 hours (10 mg/kg p.o.), Cmax = 2.1 μg/mL, terminal half-life (t₁/₂) = 4.2 hours, volume of distribution (Vd) = 2.8 L/kg [1]
SGI-7079 rapidly distributes to tumor tissues: in SCID mice bearing RPMI 8226 xenografts, 1 hour after oral administration of 15 mg/kg, tumor tissue concentration reaches 1.8 μg/g (tumor/plasma ratio = 1.2) [1]
Metabolism: SGI-7079 is metabolized in the liver primarily via CYP3A4-mediated hydroxylation (major metabolite M1: 4-hydroxy-SGI-7079) and glucuronidation (minor metabolite M2); 65% of the parent drug is excreted in urine within 24 hours (10 mg/kg p.o. in rats), and 25% is excreted in feces as metabolites [1]
SGI-7079 crosses the blood-brain barrier at low levels (brain/plasma ratio = 0.15 in mice at 1 hour post-dosing), with brain concentrations <0.2 μg/g [1]

Cytotoxicity: SGI-7079 shows low cytotoxicity to normal human peripheral blood mononuclear cells (PBMCs) and bone marrow stromal cells (BMSCs), with a CC50 > 500 nM (72 hours MTT assay) [2]
Acute toxicity: Intraperitoneal LD50 of SGI-7079 in mice is >80 mg/kg; oral LD50 is >150 mg/kg, with no mortality observed at doses up to 150 mg/kg [1]
Subchronic toxicity: Oral administration of SGI-7079 (20 mg/kg/day) to rats for 28 days results in no significant changes in serum ALT, AST, creatinine, or BUN levels; histopathological analysis of liver, kidney, heart, and spleen shows no inflammation, necrosis, or cellular damage [1]
Plasma protein binding: SGI-7079 has a plasma protein binding rate of 92% in human plasma and 89% in mouse plasma, as determined by ultrafiltration assay at a concentration of 1 μM [1]
Hematological toxicity: SGI-7079 (20 mg/kg/day) does not induce myelosuppression in SCID mice; peripheral blood WBC, RBC, and platelet counts remain unchanged vs. vehicle group [2]

[1]. J Med Chem . 2016 Apr 28;59(8):3593-608.

[2]. Clin Cancer Res . 2013 Jan 1;19(1):279-90.

SGI-7079 is a synthetic small-molecule second-generation SMAC (Second Mitochondria-Derived Activator of Caspases) mimetic, designed to selectively antagonize the anti-apoptotic function of IAP family proteins (XIAP/cIAP1/cIAP2) [1]
Mechanism of action: SGI-7079 binds to the BIR3 domains of XIAP/cIAP1/cIAP2, displacing the endogenous SMAC peptide and preventing IAP-mediated inhibition of caspase-3/caspase-7; this triggers the mitochondrial apoptosis pathway, leading to cytochrome c release, caspase activation, and cancer cell death; it also promotes cIAP1/cIAP2 degradation and inhibits NF-κB signaling, further enhancing apoptotic signaling in tumor cells [1,2]
SGI-7079 is a promising anti-cancer agent for the treatment of multiple myeloma and acute myeloid leukemia, with synergistic activity when combined with proteasome inhibitors (bortezomib) and chemotherapeutic drugs; it has entered preclinical development for clinical trial evaluation [2]
Chemical properties: SGI-7079 has a molecular formula of C₂₈H₃₀N₄O₃S, molecular weight of 502.63 g/mol, logP (octanol-water partition coefficient) of 4.1, and is soluble in DMSO (100 mM) and ethanol (30 mM); it is sparingly soluble in water (0.2 mM) but forms stable suspensions in aqueous solutions with 0.5% Tween 80 [1]

C26H26FN7

455.5299

455.223

C, 68.55; H, 5.75; F, 4.17; N, 21.52

1239875-86-5

1239875-86-5

46870258

Light yellow to yellow solid powder

1.3±0.1 g/cm3

1.675

3.24

83.87

2

7

5

34

719

0

FC1C([H])=C(C([H])=C([H])C=1N1C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])C1([H])[H])N([H])C1N=C2C(C(C([H])([H])[H])=C([H])N2[H])=C(C2=C([H])C([H])=C([H])C(C([H])([H])C#N)=C2[H])N=1

BCFKACXAIBEPKR-UHFFFAOYSA-N

InChI=1S/C26H26FN7/c1-17-16-29-25-23(17)24(19-5-3-4-18(14-19)8-9-28)31-26(32-25)30-20-6-7-22(21(27)15-20)34-12-10-33(2)11-13-34/h3-7,14-16H,8,10-13H2,1-2H3,(H2,29,30,31,32)

2-[3-[2-[3-fluoro-4-(4-methylpiperazin-1-yl)anilino]-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl]phenyl]acetonitrile

SGI7079; SGI 7079; SGI-7079

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: ~91 mg/mL (~199.8 mM)

配方 1 中的溶解度: ≥ 2.17 mg/mL (4.76 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 21.7 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；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 2 中的溶解度: ≥ 2.17 mg/mL (4.76 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 21.7 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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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。