Novel Biosynthethic Pathway for Secosteroids and the Skin

用于 Secosteroids 和皮肤的新型生物合成途径

• 批准号: 8504632
• 负责人: ARNOLD E POSTLETHWAITE
• 金额: $ 30万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8504632
• 关键词: 7-dehydrocholesterol; 7-dehydropregnenolone; Adjustment Disorders; Adjuvant; Adrenal Glands; Anti-Inflammatory Agents; Anti-inflammatory; Basic Science; Biochemical; Biodistribution; Bioinformatics; Biological; Biological Assay; Biological Testing; Biology; Bleomycin; CYP11A1 gene; CYP27B1 gene; Calcitriol; Catalytic Domain; Cell Lineage; Cell Nucleus; Cells; Cholecalciferol; Cholesterol Monooxygenase (Side-Chain-Cleaving); Chronic; Clinical; Clinical Trials; Complement; Dermal; Development; Diffuse; Disease; Dose; Drug Kinetics; Endocrinology; Environment; Enzymes; Ergocalciferols; Fibroblasts; Fibrosis; Gene Expression; Gene Silencing; Goals; Human; Hydroxylation; Inflammatory; Ligands; Metabolic Pathway; Microarray Analysis; Molecular; Mus; Oral; Organ; Orphan; Pathway interactions; Patients; Pharmacotherapy; Phase I Clinical Trials; Placenta; Positioning Attribute; Pre-Clinical Model; Production; Property; RNA Interference; Relative (related person); Rodent; Route; Schedule; Scleroderma; Secosteroids; Serum; Side; Signal Pathway; Skin; Steroids; Technology; Testing; Therapeutic; Therapeutic Agents; Tissues; Toxic effect; Vitamin D; Vitamin D3 Receptor; analog; base; cancer therapy; clinical application; comparative efficacy; design; extracellular; in vitro activity; in vivo; keratinocyte; novel; pre-clinical; public health relevance; receptor; skin disorder; trait; ultraviolet

DESCRIPTION (provided by applicant): The discovery of new secosteroidogenic pathways, regulated by P450scc represents a fundamental and unexpected advance in skin biology, endocrinology, and pharmacotherapy of vitamin D3. The novel secosteroids can serve as therapeutic agents for proliferative, fibrosing and other skin diseases, because of their high biological potency [similar to 1,25(OH)2D3] without identifiable toxic effects. Their production by P450scc in placenta, adrenals and in epidermal keratinocytes indicates that they can be produced in vivo. In fact, we detected 20(OH)D3 in human serum. This defines them as novel endogenous bioregulators. They are at least as potent as 1,25(OH)2D3 (calcitriol) in skin cells displaying anti-proliferative, pro-differentiation and anti-inflammatory properties. Important from clinical point of view, some of the P450scc-derived compounds [20(OH)D3 or 17,20(OH)2pD] are non-toxic in rodents at, respectively, extremely high doses as 30 or 3 ¿g/kg, defining them as potential therapeutic agents for fibrosing or skin hyperproliferative or inflammatory disorders, or as adjuvants in cancer therapy. To study these pathways following aims are designed: 1. Defining the mechanism of action of 20(OH)D3 and 17,20(OH)2pD in skin fibroblasts; 2. Establishing novel secosteroids (20(OH)D3 and 17,20(OH)2pD) as excellent candidates for therapy of human fibrosing diseases in preclinical models of scleroderma; 3. Evaluate dosing schedules and pharmacokinetics for 20(OH)D3, or 17,20(OH)2D3 by gavage that result in optimal suppression of murine scleroderma without causing toxicity. The aim 1 will focus on the basic science and therefore will include several subaims. Subaim 1 will investigate how signaling pathways in dermal fibroblasts are inhibited by 20(OH)D3 and 17,20(OH)2pD. Subaim 2 will investigate the degree of involvement of VDR-dependent pathways activated by 20(OH)D3 and 17,20(OH)2pD on fibroblasts. These will be complemented by testing of ligand-induced VDR translocation to the nucleus and activation of VDRE transcriptional activity. Subaim 3 will test the involvement of ROR in fibroblasts from ROR¿-/- and ROR?- /- mice with confirmation in human fibroblasts with receptors silenced by RNAi. These will be supported by biochemical and cell-based assays. Divergence and overlaps between the actions on VDR, ROR¿ and ROR? will be accomplished by microarray technology. Subaim 4 will test whether anti-fibrotic activity of novel secosteroids is potentiated by hydroxylation at C1¿ using CYP27B1-/- mice. These will be complemented by comparing phenotypic effects of 1,20(OH)2D3 and 1,17,20(OH)3pD with the parental compounds. Aim 2 will test the biological efficacy of 20(OH)D3 and 17,20(OH)2pD in preclinical models of scleroderma and will compare their i. p. to oral routes of delivery. Aim 3 is a logical continuation of aims 1 and 2 and will test their biodistribution in organ/tissue specific accumulation of the analogs and define their possible toxicity for extremely high oral doses. These are necessary for further development in Phase I clinical trials in patients with scleroderma or other fibrosing diseases, which represents our ultimate goal (clinical application).

描述（由申请人提供）：由 P450scc 调节的新的分泌类固醇生成途径的发现代表了皮肤生物学、内分泌学和维生素 D3 药物治疗方面的根本性和意想不到的进展。这种新型的类固醇可以作为增殖性、纤维化和其他皮肤病的治疗剂，因为它们具有高生物效力[类似于 1,25(OH)2D3]，且没有可识别的毒性作用。他们的生产由 胎盘、肾上腺和表皮角质形成细胞中的 P450scc 表明它们可以在体内产生。事实上，我们在人血清中检测到了 20(OH)D3。这将它们定义为新型内源性生物调节剂。它们在皮肤细胞中至少与 1,25(OH)2D3（骨化三醇）一样有效，具有抗增殖、促分化和抗炎特性。重要来自 从临床角度来看，一些 P450scc 衍生化合物 [20(OH)D3 或 17,20(OH)2pD] 在啮齿动物中分别在 30 或 3 µg/kg 的极高剂量下是无毒的，将它们定义为纤维化或皮肤过度增殖或炎症性疾病的潜在治疗剂， 或作为癌症治疗的佐剂。为了研究这些途径，设计了以下目标： 1. 确定 20(OH)D3 和 17,20(OH)2pD 在皮肤成纤维细胞中的作用机制； 2. 建立新型分泌类固醇（20(OH)D3 和 17,20(OH)2pD）作为硬皮病临床前模型中人类纤维化疾病治疗的优秀候选药物； 3. 通过灌胃法评估 20(OH)D3 或 17,20(OH)2D3 的给药方案和药代动力学，从而实现对小鼠硬皮病的最佳抑制而不引起毒性。目标 1 将侧重于基础科学，因此将包括几个子目标。 Subaim 1 将研究 20(OH)D3 和 17,20(OH)2pD 如何抑制真皮成纤维细胞中的信号传导途径。 Subaim 2 将研究 20(OH)D3 和 17,20(OH)2pD 对成纤维细胞激活的 VDR 依赖性途径的参与程度。这些将通过测试配体诱导的 VDR 易位到细胞核和 VDRE 转录活性的激活来补充。 Subaim 3将测试ROR在ROR¿-/-和ROR?-/-小鼠成纤维细胞中的参与情况，并在受体被RNAi沉默的人类成纤维细胞中进行确认。这些将得到生化和细胞检测的支持。 VDR、ROR¿ 和 ROR 行动之间的分歧和重叠？将通过微阵列技术来实现。 Subaim 4 将使用 CYP27B1-/- 小鼠测试 C1¿ 羟基化是否会增强新型 secicosteroids 的抗纤维化活性。这些将通过比较 1,20(OH)2D3 和 1,17,20(OH)3pD 与母体化合物的表型效应来补充。目标 2 将测试 20(OH)D3 和 17,20(OH)2pD 在硬皮病临床前模型中的生物学功效，并比较它们的 i。 p。口服给药途径。目标 3 是目标 1 和 2 的逻辑延续，将测试它们在类似物的器官/组织特异性积累中的生物分布，并确定它们在极高口服剂量下可能的毒性。这些对于硬皮病或其他纤维化疾病患者的 I 期临床试验的进一步发展是必要的，这代表了我们的最终目标（临床应用）。

项目成果

Pathogenesis of Systemic Sclerosis.

• DOI: 10.3389/fimmu.2015.00272
• 发表时间: 2015
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Pattanaik D;Brown M;Postlethwaite BC;Postlethwaite AE
• 通讯作者: Postlethwaite AE

Ultraviolet radiation regulates cortisol activity in a waveband-dependent manner in human skin ex vivo.

• DOI: 10.1111/bjd.12096
• 发表时间: 2013-03
• 期刊: The British journal of dermatology
• 作者: Skobowiat C;Sayre RM;Dowdy JC;Slominski AT
• 通讯作者: Slominski AT

Metabolism of 1alpha-hydroxyvitamin D3 by cytochrome P450scc to biologically active 1alpha,20-dihydroxyvitamin D3.

• DOI: 10.1016/j.jsbmb.2008.10.005
• 发表时间: 2008-12
• 期刊: JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY
• 影响因子: 4.1
• 作者: Tuckey, Robert C.;Janjetovic, Zorica;Li, Wei;Nguyen, Minh N.;Zmijewski, Michal A.;Zjawiony, Jordan;Slominski, Andrzej
• 通讯作者: Slominski, Andrzej

Expression of Vitamin D Receptor (VDR) Positively Correlates with Survival of Urothelial Bladder Cancer Patients.

维生素D受体（VDR）的表达与尿路上皮膀胱癌患者的存活正相关。

• DOI: 10.3390/ijms161024369
• 发表时间: 2015-10-15
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Jóźwicki W;Brożyna AA;Siekiera J;Slominski AT
• 通讯作者: Slominski AT

Metabolism of cholesterol, vitamin D3 and 20-hydroxyvitamin D3 incorporated into phospholipid vesicles by human CYP27A1.

• DOI: 10.1016/j.jsbmb.2011.11.012
• 发表时间: 2012-04
• 期刊: JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY
• 影响因子: 4.1
• 作者: Tieu, Elaine W;Li, Wei;Chen, Jianjun;Baldisseri, Donna M;Slominski, Andrzej T;Tuckey, Robert C
• 通讯作者: Tuckey, Robert C