Department of Dermatology and Wound Healing Centre, Copenhagen University Hospital - Bispebjerg and Frederiksberg, Copenhagen, Denmark.
Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg, Denmark.
Lasers Surg Med. 2022 Jan;54(1):170-181. doi: 10.1002/lsm.23504. Epub 2021 Dec 2.
Current cancer immunotherapeutic treatment with PD-1 inhibitors is administered systemically. However, a local treatment strategy may be advantageous as it could provide targeted drug delivery as well as attenuate side effects seen with systemic treatments. For keratinocyte cancers, where surgical excision is not always applicable, an alternate local treatment approach would be beneficial. This study aims to examine cutaneous pharmacokinetics and biodistribution of the PD-1 inhibitor nivolumab, locally delivered either by ablative fractional laser (AFL)-assisted passive diffusion or active intradermal injection, in vivo.
In vivo pig skin was either exposed to CO AFL (80 mJ/mb by two stacked pulses of 40 mJ/mb) at 5% or 15% density followed by topical application of nivolumab (1 mg/ml, 100 µl/10 × 10 mm) or intradermally injected with nivolumab (1 mg/ml, 100 µl). Cutaneous nivolumab delivery was evaluated at different timepoints (0, 1, 2, 4 hours and 2 days) at two tissue depths (100-800 and 900-1600 µm) by ELISA. Visualization of cutaneous biodistribution was shown in vertical tissue sections using HiLyte Fluor 488 SE labeled nivolumab for fluorescence microscopy whereas nivolumab was DOTA-tagged with Dysprosium before the laser ablation-inductively coupled plasma-mass spectrometry analysis (LA-ICP-MS).
Our in vivo study revealed different pharmacokinetic and biodistribution patterns for the AFL- and injection techniques. A superficial horizontal band-like uptake of nivolumab was provided with AFL-assisted passive diffusion whereas a deep focal deposition was seen with active intradermal injection, compared with controls showing remnant deposition on the skin surface. AFL-assisted nivolumab uptake in upper dermis peaked after 4 hours (p < 0.01). The cutaneous concentration of nivolumab achieved by intradermal injection was markedly higher than with AFL, the highest deposition with intradermal injection was detected at time 0 hours in both upper and deep dermis (p < 0.01) and decreased throughout the study period, although the concentration remained higher compared with saline control injections at all time points (0 hours -2 d) (p < 0.01).
Local cutaneous delivery of nivolumab with either AFL or intradermal injection revealed two different pharmacokinetic and biodistribution patterns. Passive AFL-assisted diffusion of nivolumab resulted in enhanced uptake after 4 hours, while intradermal actively injected nivolumab showed immediate enhanced cutaneous deposition with retention up to 2 days after injection. The two local delivery techniques show potential for development of individualized treatment strategies depending on the clinical tumor appearance.
目前，癌症免疫治疗中的 PD-1 抑制剂是全身性给药。然而，局部治疗策略可能具有优势，因为它不仅可以提供靶向药物递送，还可以减轻全身性治疗的副作用。对于角化细胞癌，手术切除并不总是适用，因此需要一种替代的局部治疗方法。本研究旨在探讨 PD-1 抑制剂纳武单抗的皮肤药代动力学和生物分布，通过消融性微射流（AFL）辅助被动扩散或主动皮内注射两种方法进行局部给药，进行体内实验。
体内猪皮分别以 5%或 15%密度接受 CO AFL（两次 40mJ/mb 的脉冲，每个脉冲 40mJ/mb）照射，然后进行纳武单抗（1mg/ml，100µl/10×10mm）的局部应用或皮内注射纳武单抗（1mg/ml，100µl）。通过 ELISA 在不同时间点（0、1、2、4 小时和 2 天）和两个组织深度（100-800 和 900-1600µm）评估皮肤中纳武单抗的递送情况。使用 HiLyte Fluor 488 SE 标记的纳武单抗对垂直组织切片进行皮肤生物分布可视化，而纳武单抗在激光烧蚀-电感耦合等离子体质谱分析（LA-ICP-MS）之前用 Dysprosium 进行 DOTA 标记。
我们的体内研究揭示了 AFL 和注射技术的不同药代动力学和生物分布模式。AFL 辅助被动扩散提供了浅表水平的纳武单抗摄取，而主动皮内注射则显示出深部焦点沉积，与对照组相比，对照组在皮肤表面仍有残留沉积。AFL 辅助纳武单抗在上皮真皮中的摄取在 4 小时后达到峰值（p<0.01）。皮内注射的纳武单抗皮肤浓度明显高于 AFL，在皮内注射的最高沉积在 0 小时时在上皮和深部真皮中均被检测到（p<0.01），并在整个研究期间逐渐下降，尽管与所有时间点的生理盐水对照注射相比，浓度仍较高（0 小时-2 天）（p<0.01）。
通过 AFL 或皮内注射进行局部皮肤递送纳武单抗，揭示了两种不同的药代动力学和生物分布模式。纳武单抗的 AFL 辅助被动扩散在 4 小时后导致摄取增强，而皮内主动注射的纳武单抗在注射后立即显示出增强的皮肤沉积，持续 2 天。这两种局部给药技术具有根据临床肿瘤外观制定个体化治疗策略的潜力。
