AR-12 Inhibits Multiple Chaperones Concomitant With Stimulating Autophagosome Formation Collectively Preventing Virus Replication

We have recently demonstrated that AR-12 (OSU-03012) reduces the function and ATPase activities of multiple HSP90 and HSP70 family chaperones. Combined knock down of chaperones or AR-12 treatment acted to reduce the expression of virus receptors and essential glucosidase proteins. Combined knock down of chaperones or AR-12 treatment inactivated mTOR and elevated ATG13 S318 phosphorylation concomitant with inducing an endoplasmic reticulum stress response that in an eIF2α-dependent fashion increased Beclin1 and LC3 expression and autophagosome formation. Over-expression of chaperones prevented the reduction in receptor/glucosidase expression, mTOR inactivation, the ER stress response, and autophagosome formation. AR-12 reduced the reproduction of viruses including Mumps, Influenza, Measles, Junín, Rubella, HIV (wild type and protease resistant), and Ebola, an effect replicated by knock down of multiple chaperone proteins. AR-12-stimulated the co-localization of Influenza, EBV and HIV virus proteins with LC3 in autophagosomes and reduced viral protein association with the chaperones HSP90, HSP70, and GRP78. Knock down of Beclin1 suppressed drug-induced autophagosome formation and reduced the anti-viral protection afforded by AR-12. In an animal model of hemorrhagic fever virus, a transient exposure of animals to low doses of AR-12 doubled animal survival from ∼30% to ∼60% and suppressed liver damage as measured by ATL, GGT and LDH release. Thus through inhibition of chaperone protein functions; reducing the production, stability and processing of viral proteins; and stimulating autophagosome formation/viral protein degradation, AR-12 acts as a broad-specificity anti-viral drug in vitro and in vivo. We argue future patient studies with AR-12 are warranted

Phase I study of onapristone, a type I antiprogestin, in female patients with previously treated recurrent or metastatic progesterone receptor-expressing cancers.

INTRODUCTION:Onapristone is a type I progesterone receptor (PR) antagonist, which prevents PR- mediated DNA transcription. Onapristone is active in multiple preclinical models and two prior studies demonstrated promising activity in patients with breast cancer. We conducted a study of extended release (ER) Onapristone to determine a recommended dose and explore the role of transcriptionally-activated PR (APR), detected as an aggregated subnuclear distribution pattern, as a predictive biomarker. METHODS:An open-label, multicenter, randomized, parallel-group, phase 1 study (target n = 60; NCT02052128) included female patients ≥18 years with PRpos tumors. APR analysis was performed on archival tumor tissue. Patients were randomized to five cohorts of extended release (ER) onapristone tablets 10, 20, 30, 40 or 50 mg BID, or immediate release 100 mg QD until progressive disease or intolerability. Primary endpoint was to identify the recommended phase 2 dose. Secondary endpoints included safety, clinical benefit and pharmacokinetics. RESULTS:The phase 1 dose escalation component of the study is complete (n = 52). Tumor diagnosis included: endometrial carcinoma 12; breast cancer 20; ovarian cancer 13; other 7. Median age was 64 (36-84). No dose limiting toxicity was observed with reported liver function test elevation related only to liver metastases. The RP2D was 50 mg ER BID. Median therapy duration was 8 weeks (range 2-44), and 9 patients had clinical benefit ≥24 weeks, including 2 patients with APRpos endometrial carcinoma. CONCLUSION:Clinical benefit with excellent tolerance was seen in heavily pretreated patients with endometrial, ovarian and breast cancer. The data support the development of Onapristone in endometrial endometrioid cancer. Onapristone should also be evaluated in ovarian and breast cancers along with APR immunohistochemistry validation

Evaluation of Functional Stability and Batch-to-Batch Reproducibility of a Castanea sativa Leaf Extract with Antioxidant Activity

A growing body of evidence suggests that free radicals are generated by UV irradiation being responsible for skin injury. In this regard, the topical use of formulations composed of plant extracts with antioxidant activity could represent a useful strategy for the prevention of photoaging and oxidative-stress-mediated diseases. The aim of this study was to assess the reproducibility of the extraction method and the functional stability of a Castanea sativa leaf extract in view of its application as topical antioxidant. Measurements of 1,1-diphenyl-2-picryl hydrazyl (DPPH) scavenging activity, total phenols (measured by the Folin Ciocalteu assay) and phenolic composition (high-performance liquid chromatography unit coupled to a UV detector) were carried out on three different batches. The influence of pH and temperature on the extract’s DPPH scavenging activity was assessed in aqueous and glyceric solutions (0.025% w/v) over a 3-month period. Minor differences were found between the three extract batches for all the evaluated parameters, and therefore the reproducibility of the extraction method can be inferred. pH presented a great influence in the extract functional stability. Major antioxidant activity decrease was found at pH 7.1, while lower changes were observed at pH 5. Glyceric solutions were stable throughout the test period. At 40°C and pH 5, a marked decrease of activity was observed. Again, glyceric solutions were the most stable, even at 40°C. Proper selection of pH and solvent is mandatory to ensure the stability of the studied extract after being incorporated in semisolid forms. In view of these results, glycerine is proposed as the best vehicle for topical formulations incorporating C. sativa leaf extract, which should have a pH around 5