Bhagavathula Narasimharao, Nerusu Kamalakar C, Hanosh Andrew, Aslam Muhammad N, Sundberg Thomas B, Opipari Anthony W, Johnson Kent, Kang Sewon, Glick Gary D, Varani James
Department of Pathology, The University of Michigan, 1301 Catherine Rd., Box 0602, Ann Arbor, MI 48109, USA.
J Pharmacol Exp Ther. 2008 Mar;324(3):938-47. doi: 10.1124/jpet.107.130955. Epub 2007 Nov 30.
7-Chloro-5-(4-hydroxyphenyl)-1-methyl-3-(naphthalen-2-ylmethyl)-4,5-dihydro-1H-benzo[b][1,4]diazepin-2(3H)-one (Bz-423) is a benzodiazepine that has cytotoxic and cytostatic activity against a variety of cells in vivo and in vitro. In the present study, we demonstrate that Bz-423 (formulated for topical delivery) reduces epidermal hyperplasia in human psoriatic skin after transplantation to severe, combined immunodeficient (scid) mice. Bz-423 also suppresses the hyperplasia that develops in nonpsoriatic human skin as a consequence of transplantation to scid mice. Proliferation of human epidermal keratinocytes in monolayer culture was suppressed by Bz-423 at concentrations of 0.5 to 2.0 muM (noncytotoxic concentrations). Keratinocyte growth inhibition was accompanied by increased oxidant generation in Bz-423-treated cells, and treatment with vitamin E along with Bz-423 reversed the growth inhibition. Growth inhibition was accompanied by a redistribution of beta-catenin from a cytoplasmic pool to the cell membrane and by reduced levels of c-myc and cyclin D1 (two molecules associated with Wnt pathway signaling). Several analogs of Bz-423 were examined for antiproliferative activity against human epidermal keratinocytes and human dermal fibroblasts in monolayer culture. Each of the analogs tested suppressed growth of both cell types, but in all cases, keratinocytes were more sensitive than fibroblasts. Two of the compounds were found to suppress epidermal hyperplasia induced with all-trans retinoic acid in organ cultures of human skin. Taken together, these data show that Bz-423 and certain analogs produce biological responses in skin cells in vitro and in vivo that are consistent with therapeutic goals for treating psoriasis or epidermal hyperplasia resulting from other causes.
7-氯-5-(4-羟基苯基)-1-甲基-3-(萘-2-基甲基)-4,5-二氢-1H-苯并[b][1,4]二氮杂卓-2(3H)-酮(Bz-423)是一种苯二氮卓类药物,在体内和体外对多种细胞具有细胞毒性和细胞生长抑制活性。在本研究中,我们证明Bz-423(配制用于局部给药)在移植到严重联合免疫缺陷(scid)小鼠后,可减少人银屑病皮肤中的表皮增生。Bz-423还可抑制因移植到scid小鼠而在非银屑病人类皮肤中发生的增生。在单层培养中,浓度为0.5至2.0μM(非细胞毒性浓度)的Bz-423可抑制人表皮角质形成细胞的增殖。角质形成细胞生长抑制伴随着Bz-423处理的细胞中氧化剂生成增加,同时用维生素E与Bz-423一起处理可逆转生长抑制。生长抑制伴随着β-连环蛋白从细胞质池重新分布到细胞膜,并伴随着c-myc和细胞周期蛋白D1(与Wnt通路信号传导相关的两种分子)水平降低。研究了Bz-423的几种类似物对单层培养中的人表皮角质形成细胞和人真皮成纤维细胞的抗增殖活性。所测试的每种类似物均抑制两种细胞类型的生长,但在所有情况下,角质形成细胞比成纤维细胞更敏感。发现其中两种化合物可抑制人皮肤器官培养中全反式维甲酸诱导的表皮增生。综上所述,这些数据表明Bz-423和某些类似物在体外和体内的皮肤细胞中产生的生物学反应与治疗银屑病或其他原因引起的表皮增生的治疗目标一致。