Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan.
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan; H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan.
Eur J Pharmacol. 2021 Jan 15;891:173700. doi: 10.1016/j.ejphar.2020.173700. Epub 2020 Nov 1.
Pharmacologically induced production of fetal hemoglobin (HbF) is a pragmatic therapeutic strategy for the reduction of globin chain imbalance and improving the clinical severities of patients with β-hemoglobinopathies. To identify highly desirable new therapeutic HbF-inducing agents, we screened functionally diverse ten monoterpenes, as molecular entities for their potent induction and erythroid differentiation ability in human erythroleukemia cell line (K562) and transgenic mice. Benzidine hemoglobin staining demonstrated six compounds to have significantly induced erythroid differentiation of K562 cells in a dose and time-dependent manner. This induction paralleled well with the optimal accumulated quantity of total hemoglobin in treated cultures. The cytotoxic studies revealed that three (carvacrol, 3-carene, and 1,4-cineole) of the six compounds with their maximal erythroid expansion ability did not affect cell proliferation and were found non-toxic. Four compounds were found to have high potency, with 4-8-fold induction of HbF at both transcriptional and protein levels in vitro. Subsequently, an in vivo study with the three active non-cytotoxic compounds showed significant overexpression of the γ-globin gene and HbF production. Carvacrol emerged as a lead HbF regulator suggested by the increase in expression of γ-globin mRNA content (5.762 ± 0.54-fold in K562 cells and 5.59 ± 0.20-fold increase in transgenic mice), accompanied by an increase in fetal hemoglobin (F-cells) levels (83.47% in K562 cells and 79.6% in mice model). This study implicates monoterpenes as new HbF inducing candidates but warrants mechanistic elucidation to develop them into potential therapeutic drugs in β-thalassemia and sickle cell anemia.
药理诱导胎儿血红蛋白（HbF）的产生是一种实用的治疗策略，可以减少球蛋白链失衡，改善β-血红蛋白病患者的临床严重程度。为了鉴定具有吸引力的新的 HbF 诱导剂，我们筛选了十种具有不同功能的单萜，作为潜在的分子实体，研究其在人红白血病细胞系（K562）和转基因小鼠中的强烈诱导 HbF 产生和红细胞分化能力。联苯胺血红蛋白染色显示，有六种化合物以剂量和时间依赖的方式显著诱导 K562 细胞的红细胞分化。这种诱导与处理培养物中总血红蛋白的最佳积累量很好地平行。细胞毒性研究表明，在具有最大红细胞扩增能力的六种化合物中，有三种（香芹酚、3-蒈烯和 1,4-桉树脑）化合物不影响细胞增殖，且无细胞毒性。有四种化合物具有高活性，在体外可使 HbF 在转录和蛋白水平上分别诱导 4 至 8 倍。随后，对三种活性非细胞毒性化合物的体内研究表明，γ-珠蛋白基因和 HbF 产生显著过表达。香芹酚通过增加 γ-珠蛋白 mRNA 含量（在 K562 细胞中为 5.762 ± 0.54 倍，在转基因小鼠中为 5.59 ± 0.20 倍）和增加胎儿血红蛋白（F-细胞）水平（在 K562 细胞中为 83.47%，在小鼠模型中为 79.6%），成为 HbF 调节因子的候选者。本研究表明单萜类化合物是新的 HbF 诱导剂，但需要进一步阐明其机制，以将其开发成β-地中海贫血和镰状细胞贫血的潜在治疗药物。
