We are currently developing zilucoplan for subcutanous, or SC, self-administration. Unsatisfactory drug availability due to problems relating to this route of administration or the ability of the drug to bind to its target is another potential cause of lack of efficacy of zilucoplan if and when it is commercialized. C5, the target of zilucoplan is predominantly found in blood. For gMG and PNH,

“The possible formation of nitrosamines from nitrites and secondary amines in acidic conditions was already well-known to the food industry,” says Andre. “The use of sodium nitrite should have been a red flag prompting a check of possible presence of secondary amines, but it was not,” he says.

To determine if the effect of cholesterol modulation on HCN1 channels could be generalized to other human HCN channel isoforms, we further examined the effects on the other cardiac isoforms, HCN2 and HCN4 (Figs 2 and 3). Intriguingly, we observed differential effects of cholesterol modulation on these isoforms. Similar to HCN1, both HCN2 and HCN4 channels showed a decrease in current density upon cholesterol depletion by either MβCD (Fig. 2A,B; Fig. 3A,B) or mevastatin (Supp. Fig. 1B,C). Moreover, current densities in cells expressing these isoforms enriched with cholesterol remained similar to control (Fig. 2A,B; Fig. 3A,B). HCN2 channels showed no differences in steady-state activation properties with changes in cholesterol content, however, steady-state properties of HCN4 channels were shifted approximately +10 mV by either cholesterol depletion or enrichment (Fig. 3C; Table 1). Tail currents were too small in HCN4 expressing cells treated with mevastatin to reliably enable us to determine steady-state activation properties. Intriguingly, the effects of cholesterol modulation on human HCN2 and HCN4 kinetics differed from our observations in human HCN1 channels. The activation kinetics of HCN2 and HCN4 channels were unaffected by cholesterol modulation (Figs 2D and 3D,E). However, unlike HCN1 channels, the deactivation kinetics of HCN2 and HCN4 channels were slowed by cholesterol enrichment (Figs 2E and 3F). These data suggest the effect of cholesterol on HCN channels is isoform specific.

Galena Biopharma Inc. (NasdaqCM:GALE) is a Portland, Oregon-based biopharmaceutical company that develops innovative, targeted oncology treatments that address major unmet medical needs to advance cancer care.

Natural killer (NK) cells are essential for immunosurveillance against transformed cells. Transient receptor potential melastatin 2 (TRPM2) is a Ca2+-permeable cation channel gated by ADP-ribose (ADPR). However, the role of TRPM2-mediated Ca2+ signaling in the antitumor response of NK cells has not been explored. Here, we show that ADPR-mediated Ca2+ signaling is important for cytolytic granule polarization and degranulation but not involved in target cell recognition by NK cells. The key steps of this pathway are: 1) the activation of intracellular CD38 by protein kinase A following the interaction of the NK cell with a tumor cell results in the production of ADPR, 2) ADPR targets TRPM2 channels on cytolytic granules, and 3) TRPM2-mediated Ca2+ signaling induces cytolytic granule polarization and degranulation, resulting in antitumor activity. NK cells treated with 8-Br-ADPR, an ADPR antagonist, as well as NK cells from Cd38−/− mice showed reduced tumor-induced granule polarization, degranulation, granzyme B secretion, and cytotoxicity of NK cells. Furthermore, TRPM2-deficient NK cells showed an intrinsic defect in tumoricidal activity. These results highlight CD38, ADPR, and TRPM2 as key players in the specialized Ca2+ signaling system involved in the antitumor activity of NK cells.

Market exclusivity provisions under the FDCA also can delay the submission or the approval of certain applications. The FDCA provides a five-year period of non-patent marketing exclusivity within the United States to the first applicant to gain approval of an NDA for a new chemical entity. A drug is a new chemical entity if the FDA has not previously approved any other new drug containing the same active moiety, which is the molecule or ion responsible for the action of the drug substance. During the exclusivity period, the FDA may not accept for review an abbreviated new drug application, or ANDA, or a 505(b)(2) NDA submitted by another company for another version of such drug where the applicant does not own or have a legal right of reference to all the data required for approval. However, an application may be submitted after four years if it contains a certification of patent invalidity or non-infringement. The FDCA also provides three years of marketing exclusivity for an NDA, 505(b)(2) NDA or supplement to an approved NDA if new clinical investigations, other than bioavailability studies, that were conducted or sponsored by the applicant are deemed by the FDA to be essential to the approval of the application, for example, for new indications, dosages or strengths of an existing drug. This three-year exclusivity covers only the conditions associated with the new clinical investigations and does not prohibit the FDA from approving ANDAs for drugs containing the original active agent. Five-year and three-year exclusivity will not delay the submission or approval of a full NDA; however, an applicant submitting a full NDA would be required to conduct or obtain a right of reference to all of the pre-clinical studies and adequate and well-controlled clinical trials necessary to demonstrate safety and effectiveness.

We would like to thank Dr. Juliane Stieber for her generous gift of the human HCN clones, and Dr. Rikard Blunck for access to necessary equipment. Sources of Funding: This work was supported by a Grant-in-Aid from the Heart & Stroke Foundation of Canada (Award No. G-13-0001882). OF is supported by a bourses de prestige from GÉPROM, an FRQS funded research group.

Because 8-Br-ADPR, an antagonistic analog of ADPR, inhibited the degranulation and cytolytic activity of NK cells, we investigated whether ADPR was endogenously produced in tumor cell-stimulated NK cells and if CD38 was the enzyme responsible for ADPR production. The level of ADPR in Cd38+/+ NK cells increased 2.4 fold when stimulated by PME, which was not observed in Cd38−/− NK cells (Fig. 4a). By contrast, the level of cADPR, another Ca2+ signaling second messenger produced by CD38, was not affected by stimulation with the PME of tumor cells (Fig. 4b). These results show that ADPR is specifically induced by CD38 in response to tumor cells.

If commenced, the ARMOR Study may also be terminated as a result of, but not limited to, safety signals. In addition, the ARMOR Study or other clinical trials may be suspended or terminated by us, the FDA or other regulatory authorities, the principal investigator at a site, the IRBs at the sites where such boards are overseeing a trial or the data safety monitoring board, or the DSMB, that is overseeing the clinical trial at issue, or other regulatory authorities due to a number of factors, including:

If an emerging growth company that prepares its financial statements in accordance with U.S. GAAP, indicate by check mark if the registrant has elected not to use the extended transition period for complying with any new or revised financial accounting standards provided pursuant to Section 7(a)(2)(B) of the Securities Act. x

Jin, R. C. & Loscalzo, J. Vascular Nitric Oxide: Formation and Function. J Blood Med. 2010, 147–162 (2010).

Hi Mark, Your comments about BENEFITS of subcutaneous adipose tissue of are very interesting. I consider subcutaneous adipose tissue on arms and legs neutral as opposed to visceral adipose tissue.