Gumułka Paweł, Tarsa Monika, Dąbrowska Monika, Starek Małgorzata
Department of Inorganic and Analytical Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College, 9 Medyczna St., 30-688 Kraków, Poland.
Doctorial School of Medical and Health Sciences, Jagiellonian University Medical College, 16 Łazarza St., 31-530 Kraków, Poland.
Biomedicines. 2022 Nov 5;10(11):2821. doi: 10.3390/biomedicines10112821.
Grapiprant is a new analgesic and anti-inflammatory drug belonging to the piprant class, approved in 2016 by the FDA Veterinary Medicine Center for the treatment of pain and inflammation associated with osteoarthritis in dogs. It acts as a highly selective antagonist of the EP4 receptor, one of the four prostaglandin E2 (PGE2) receptor subtypes. It has been shown to have anti-inflammatory effects in rat models of acute and chronic inflammation and clinical studies in people with osteoarthritis. The current state of knowledge suggests the possibility of using it in oncological therapy. The manuscript presents the development of conditions for the identification and quantitative determination of grapiprant by thin-layer chromatography with densitometric detection. The optimal separation of the substance occurs using silica gel 60F chromatographic plates and the mobile phase containing ethyl acetate-toluene-butylamine. Validation (according to ICH requirements) showed that the developed method is characterized by straightness of results in a wide concentration range with the limit of detection of 146.65 µg/mL. The %RSD values of the precision and accuracy confirm the sensitivity and reliability of the developed procedure. Next, the method was used for quantification of grapiprant in a pharmaceutical preparation, and for stability studies under various environmental conditions. Additionally, the mass studies were carried out on the stressed samples using the UPLC-MS/MS method. The degradation products were primarily characterized by comparing their mass fragmentation profiles with those of the drug. The results indicated a potential degradation pathway for grapiprant.
格拉匹普兰是一种新型的镇痛抗炎药物，属于匹普兰类，于2016年获美国食品药品监督管理局兽药中心批准，用于治疗犬骨关节炎相关的疼痛和炎症。它是前列腺素E2（PGE2）四种受体亚型之一的EP4受体的高选择性拮抗剂。在急性和慢性炎症大鼠模型以及骨关节炎患者的临床研究中，它已显示出抗炎作用。目前的知识状况表明有可能将其用于肿瘤治疗。该手稿介绍了采用薄层色谱-密度测定法鉴定和定量测定格拉匹普兰的条件的开发情况。使用硅胶60F色谱板和含有乙酸乙酯-甲苯-丁胺的流动相可实现该物质的最佳分离。验证（根据国际人用药品注册技术协调会要求）表明，所开发的方法在宽浓度范围内结果具有线性，检测限为146.65 µg/mL。精密度和准确度的相对标准偏差值证实了所开发方法的灵敏度和可靠性。接下来，该方法用于药物制剂中格拉匹普兰的定量分析以及各种环境条件下的稳定性研究。此外，使用超高效液相色谱-串联质谱法对加速试验样品进行了质量研究。通过将降解产物的质量碎片图谱与药物的图谱进行比较，对降解产物进行了初步表征。结果表明了格拉匹普兰的潜在降解途径。
