17α-Ethynyl-5α-androstane-3α, 17β-diol Treatment of MNU-Induced Mammary Cancer in Rats

N-methyl-N-nitrosourea (MNU) induces estrogen-dependent mammary tumors in female Lewis rats. We explored the antineoplastic activity of a synthetic androstane derivative, 17α-ethynyl-5α-androstane-3α, 17β-diol (HE3235), as a single agent or in combination with docetaxel compared to tamoxifen, anastrazole, and docetaxel monotherapies against MNU-induced mammary tumors in female Lewis rats. Treatment with HE3235 alone rapidly reduced tumor burden, similar in effect to tamoxifen and anastrozole. The combination of HE3235 with docetaxel was more effective than any single agent, although without apparent toxicity. Only HE3235 or HE3235 plus docetaxel continued to suppress tumor growth after cessation of treatment. HE3235 treatment increased immunohistochemical markers of apoptosis and expression of proapoptotic genes and estrogen receptor beta and decreased expression of antiapoptotic genes, androgen receptor, and estrogen receptor alpha. These data warrant clinical investigation of HE3235 for breast cancer treatment

5-Androstene-3β,7β,17β-triol (β-AET) Slows Thermal Injury Induced Osteopenia in Mice: Relation to Aging and Osteoporosis

5-androstene-3β,7β,17β-triol (β-AET), an active metabolite of dehydroepiandrosterone (DHEA), reversed glucocorticoid (GC)-induced suppression of IL-6, IL-8 and osteoprotegerin production by human osteoblast-like MG-63 cells and promoted osteoblast differentiation of human mesenchymal stem cells (MSCs). In a murine thermal injury model that includes glucocorticoid-induced osteopenia, β-AET significantly (p<0.05) preserved bone mineral content, restored whole body bone mineral content and endochondral growth, suggesting reversal of GC-mediated decreases in chondrocyte proliferation, maturation and osteogenesis in the growth plate. In men and women, levels of β-AET decline with age, consistent with a role for β-AET relevant to diseases associated with aging. β-AET, related compounds or synthetic derivatives may be part of effective therapeutic strategies to accelerate tissue regeneration and prevent or treat diseases associated with aging such as osteoporosis

17α-Ethynyl-androst-5-ene-3β,7β,17β-triol (HE3286) Is Neuroprotective and Reduces Motor Impairment and Neuroinflammation in a Murine MPTP Model of Parkinson’s Disease

17α-Ethynyl-androst-5-ene-3β,7β,17β-triol (HE3286) is a synthetic androstenetriol in Phase II clinical development for the treatment of inflammatory diseases. HE3286 was evaluated for blood-brain barrier (BBB) permeability in mice, and efficacy in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) murine model of Parkinson’s disease (PD). We found that HE3286 freely penetrated the BBB. HE3286 treatment significantly improved motor function compared to vehicle in the rotarod test (mean 58.2 sec versus 90.9 sec, P<0.0001), and reduced inflammatory mediator gene expression in the brain (inducible nitric oxide synthase, 20%, P=0.002; tumor necrosis factor α, 40%, P=0.038, and interleukin-1β, 33%, P=0.02) measured by reverse-transcriptase polymerase chain reaction. Brain tissue histopathology and immunohistochemistry showed that HE3286 treatment increased the numbers of tyrosine hydroxylase-positive cells by 17% compared to vehicle (P=0.003), and decreased the numbers of damaged neurons by 38% relative to vehicle (P=0.029). L-3,4-dihydroxyphenylalanine (L-DOPA) efficacy was not enhanced by concurrent administration of HE3286. HE3286 administration prior to MPTP did not enhance efficacy. Our data suggest a potential role for HE3286 in PD treatment, and provides incentive for further investigation

Effect of β-AET on human MSC differentiation.

<p>Mesenchymal stem cells (MSC) derived from human bone were cultured with β-AET (0, 0.1, 1 or 10 µM) for 15 days. The percentage of cells expressing the preosteoblast integrins osteopontin (OP) were measured by flow cytometry. Data are expressed as average % cells +/− standard deviation.</p

Effect of β-AET on cancellous bone morphometry and histomorphometry and bone resorption and growth.

<p>Male BALB/c mice (n = 10 per group) were subjected to 20% total body surface area burn and treated (sc injection) with vehicle alone, or with β-AET (25 or 50 mg/kg) immediately after thermal trauma. An identical treatment was given 48 hours later and then 3 times per week for 4 weeks. Cancellous bone morphometry and histomorphometry were measured at the proximal tibial metaphysis of mice (A, B). The endochondral growth rate was measured at the tibial epiphyseal growth plate (C) Endocortical eroded surface was measured at the surface of the mid-diaphyseal shaft of the femur (D), as an indicator of osteoclastic bone. ‘#’, ‘b’ and ‘*’ indicate significant differences from the sham control, baseline and vehicle group, respectively, <i>p</i><0.05. Data are expressed as means ± SEM.</p

Effect of β-AET on femur weights of mice subjected to thermal injury.

<p>Male BALB/c mice (n = 10 per group) were subjected to 20% total body surface area and treated (sc injection) with vehicle alone, or with β-AET (25 or 50 mg/kg) immediately after thermal trauma. An identical treatment was given 48 hours later and then 3 times per week for 4 weeks. Femur was weighed when wet (A), after drying (B) and upon ashing (C). Panel D indicates whole body BMC. ‘#’, ‘b’ and ‘*’ indicate significant difference from the sham control, baseline and vehicle group, respectively, <i>p</i><0.05. The bars represent means ± SEM.</p

Correlation of β-AET levels in human plasma with age.

<p>β-AET levels were measured in plasma samples taken from 102 males (aged 20–80) and 150 females (aged 20–73) by reverse phase LC-MS/MS. Linear regression analysis revealed significantly non-zero slope for males (<i>p</i> = 0.02; r² = 0.06) and females (<i>p</i><0.0001; r² = 0.12).</p

Effect of β-AET on GC-induced suppression of genes in MG-63 cell line.

<p>MG-63 cells pre-treated (1 h) with β-AET were exposed to Dex for 8 hours. The expression of IL-6 (A), IL-8 (B) and OPG (C) was determined by ELISA in the culture medium. Significant differences from control (0.04% DMSO) are indicated as follows: *** = (<i>p</i><0.001), ** = (<i>p</i><0.05) and * = (<i>p</i><0.01). # indicates a significant difference from Dex (<i>p</i><0.05). + indicates strong trend (<i>p</i><0.1). Data are expressed as means ± sem; n = 3. Absorbance values are plotted for IL-8 (panel B) because IL-8 concentrations extrapolated from standard curve in Dex only samples were below the limits of the standard curve and could not be reliably calculated.</p