Role of nano-lipid formulation of CARP-1 mimetic, CFM-4.17 to improve systemic exposure and response in osimertinib resistant non-small cell lung cancer.
Role of nano-lipid formulation of CARP-1 mimetic, CFM-4.17 to improve systemic exposure and response in osimertinib resistant non-small cell lung cancer.
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Carp-1 Mimetic，CFM-4.17的纳米脂质制剂在埃米默替尼耐药的非小细胞肺癌中的全身暴露和反应的作用。
DOI：
10.1016/j.ejpb.2020.11.007
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Singh M
中科院分区：
文献类型：
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作者：
Kommineni N;Nottingham E;Bagde A;Patel N;Rishi AK;Dev SRS;Singh M
EGFR突变的NSCLC已显示使用CARP-1来超越酪氨酸激酶抑制剂（如奥希替尼）的信号传导抑制。CFM 4.17是一种CARP-1抑制剂，当用作预处理时，其通过促进细胞凋亡在克服酪氨酸激酶抑制剂（TKI）抗性方面具有有前途的作用。缺乏溶解性、疏水性导致全身暴露不良是CFM 4.17的局限性。这可以通过CFM 4.17的纳米脂质制剂（NLPF）来克服，该制剂可以增强临床前动物模型中的全身暴露，并提高耐药癌细胞系的治疗效果。对CFM 4.17进行了分子对接模拟研究。CFM 4.17-NLPF通过熔融分散技术配制，并使用Design Expert和MATLAB使用Box-Behnken设计的表面响应方法学方法进行优化。在体外，CFM 4.17释放研究在模拟胃液（SGF-pH-1.2）和模拟肠液（SIF-pH-6.8）中进行。在非小细胞肺癌的2D和3D培养模型中，用HCC 827和H1975奥希替尼耐药和非耐药细胞进行细胞活力测定，以确定CFM 4.17预处理对奥希替尼应答的影响。进行大鼠体内药代动力学，测量NLPF对CFM 4.17的影响，以改善全身暴露。CFM 4.17被很好地容纳在人EGFR酪氨酸激酶活性位点的活性口袋中。采用稳健性实验设计优化CFM 4.17 NLPF，粒径小于300 nm，包封率%为92.3±1.23。在SGF和SIF中观察到CFM 4.17从NLPF的基于持续扩散的释放，分别具有基于Peppas和Higuchi的释放动力学。在非小细胞肺癌的HCC 827和H1975奥希替尼耐药和非耐药细胞的2D单层和3D球状体试验中，CFM 4.17预处理通过使IC 50值降低2倍而改善缓解。CFM4.17 NLPF与悬浮液在2D单层培养预处理中无差异，但3D培养试验表明CFM4.17 NLPF提高了组合敏感性。药代动力学分析显示，与游离CFM 4.17相比，CFM 4.17 NLPF显示出更高的AUCtot（2.9倍）和Cmax（1.18倍）。相比之下，与等效剂量混悬液相比，给予CFM 4.17 NLPF的动物组显示出4.73倍（半衰期）和3.07倍（MRT）增加。我们已经通过稳健的RSM设计方法成功地配制了CFM 4.17 NLPF，通过使细胞对奥希替尼治疗敏感以及改善CFM 4.17的口服生物利用度来显示改善的响应。
EGFR mutated NSCLCs have been shown to employ the use of CARP-1 in overriding the signaling inhibition of tyrosine kinase inhibitors (such as Osimertinib). CFM 4.17 is a CARP-1 inhibitor which has a promising role in overcoming Tyrosine Kinase Inhibitor (TKI) resistance when used as a pre-treatment through promoting apoptosis. Lack of solubility, hydrophobicity leading to poor systemic exposure are the limitations of CFM 4.17. This can be overcome by nano lipid-based formulation (NLPF) of CFM 4.17 which can enhance systemic exposure in preclinical animal models as well as improve therapeutic efficacy in drug-resistant cancer cell lines. Molecular docking simulation studies were performed for CFM 4.17. CFM 4.17-NLPF was formulated by melt dispersion technique and optimized using a Box-Behnken designed surface response methodology approach using Design Expert and MATLAB. In vitro, CFM 4.17 release studies were performed in simulated gastric fluids (SGF-pH-1.2) and simulated intestinal fluids (SIF- pH-6.8). Cell viability assays were performed with HCC827 and H1975 Osimertinib resistant and non-resistant cells in 2D and 3D culture models of Non-small cell lung cancer to determine the effects of CFM 4.17 pre-treatment in Osimertinib response. In vivo pharmacokinetics in rats were performed measuring the effects of NLPF on CFM 4.17 to improve the systemic exposure. CFM 4.17 was well accommodated in the active pocket of the active site of human EGFR tyrosine kinase. CFM 4.17 NLPF was optimized with robust experimental design with particle size less than 300 nm and % entrapment efficiency of 92.3±1.23. Sustained diffusion-based release of CFM 4.17 was observed from NLPF in SGF and SIFs with Peppas and Higuchi based release kinetics, respectively. CFM 4.17 pretreatment improved response by decreasing IC50 value by 2-fold when compared to single treatment Osimertinib in both 2D monolayer and 3D spheroid assays in HCC827 and H1975 Osimertinib resistant and non-resistant cells of Non-small cell lung cancer. There were no differences between CFM 4.17 NLPF and suspension in 2D monolayer culture pretreatments; however, The 3D culture assays showed that CFM 4.17 NLPF improved combination sensitivity. Pharmacokinetic analysis showed that CFM 4.17 NLPF displayed higher AUCtot (2.9-fold) and Cmax (1.18-fold) as compared to free CFM 4.17. In contrast, the animal groups administered CFM 4.17 NLPF showed a 4.73-fold (in half-life) and a 3.07-fold increase (in MRT) when compared to equivalent dosed suspension. We have successfully formulated CFM 4.17 NLPFs by robust RSM design approach displaying improved response through sensitizing cells to Osimertinib treatment as well as improving the oral bioavailability of CFM 4.17.