Abstract Introduction: SNX-2112 is a potent anti-tumor compound with effects on epigenetics (hypomethylation) and onco-metabolism. SNX-5422 is the orally active prodrug of SNX-2112, a highly selective inhibitor of heat-shock protein 90 (Hsp90) that has shown anti-tumor activity in clinical trials. Hsp90 is commonly elevated in tumor tissues and plays an important role in immune responses. Previous investigations have shown that SNX-2112 is highly active in TP53 null tumors and has good activity in combination with checkpoint inhibitors in syngeneic mice studies. Preclinical studies have suggested that Hsp90 inhibitors enhance antigen-specific T-cell recognition of cancer cells, increase the expression of both differentiation and MHC Class I antigens, and do not interfere with T-cell function. Methods: In two in vitro studies, the effects of SNX-2112 on immune (A375 melanoma cell line and the TP53 mutant chronic myelocytic leukemia K562 cell line) and mitochondrial genes (K652) using transcriptomics were investigated. A375 and K562 cells were cultured in media (GIBCO or Sigma, USA) supplemented with 10% fetal bovine serum at 37°C, 5% CO2, and 95% humidity. Data from transcriptomics were obtained, and affected genes analyzed. Results: In A375 cells, transcriptomics analysis revealed that SNX-2112 downregulated multiple overexpressed immune-related genes (including NFKB1, NFKB2, BRAF, HLA-DMA, HLA-DMB, HLA-DOB, HLA-DQB1, and HLA-DRB6) and upregulated multiple suppressed immune-related genes (including IFIT1, IFIT2, IFIT3, MLANA, DDX58, PMEL (gp100), DCT (TRP-2), MR1, MICA, and MICB). Transcriptomics analysis of SNX-2112 on K562 immune genes revealed that IFIT1-3, IFI6, IFI44, ISG15, DDX-58, and HLA-DRA were all upregulated. Genes downregulated by SNX-2112 included NFKB and IKBKB (Ikk), HLA-DOB, HLA-DPA1, HLA-DPB1, HLA-DQB1, PSMB8, and TAP1. In both the A375 and K562 cells lines, SNX-2112 reduced the expression of MYB oncogene, (reported to control regulatory T-cell proliferation) and caused increases in RB1CC1 (fip200) and STAT1, crucial survival factors for naïve T-cells. Additionally, tryptophan metabolism genes IDO1 and TDO2 were downregulated by SNX-2112. K562 mitochondrial genes downregulated by SNX-2112 included the following: •TP53 and p53 family members-controlled markers: HK2, GLS2, TIGAR •Glutamine addiction/Warburg effect: SLC1A5, GLUD1 •Folate pathway: SLC19A1, TYMS, SHMT1, AS3MT, DHFR2 •OXPHOS: CISD1, CHCHD4 Conclusion: SNX-2112 increased multiple suppressed immune-related genes and downregulated multiple overexpressed immune-related genes in A375 and K562 cell lines. Anti-tumor activity of SNX-2112 appears to be, in part, the result of interference with onco-metabolic pathways. Inhibition of IDO1 and TDO2 suggests possibly enhanced anti-tumor activity in combination with checkpoint inhibitors. Citation Format: Everardus Orlemans, Steven E. Hall. Transcriptomics analysis of SNX-2112 on immune and mitochodrial genes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2758.

OBJECTIVES:To establish rates of cytomegalovirus (CMV) transmission with use of CMV-unselected (CMV-U), leukocyte-reduced blood components transfused to CMV-seronegative patient/CMV-seronegative donor (CMV neg/neg) allogeneic stem cell transplantation (SCT) recipients including those receiving T-depleted grafts.BACKGROUND:CMV infection remains a major cause of morbidity following SCT. CMV-seronegative SCT recipients are particularly at risk of transfusion transmitted CMV (TT-CMV) and until recently they have received blood components from CMV-seronegative donors with significant resource implications. Although leukocyte reduction of blood components is reported to minimise risk of TT-CMV, its efficacy in high-risk situations, such as in T-depleted transplant recipients, is unknown.METHODS:We retrospectively analysed the incidence of TT-CMV in CMV neg/neg allogeneic SCT recipients transfused with CMV-U, leukocyte-reduced blood components in two transplantation centres in the UK. Patients were monitored for CMV infection by weekly CMV polymerase chain reaction testing. Leukocyte reduction of blood components was in accordance with current UK standards.RESULTS:Among 76 patients, including 59 receiving in vivo T-depletion, no episodes of CMV infection were detected. Patients were transfused with 1442 CMV-unselected, leukocyte-reduced components, equating to 1862 donor exposures.CONCLUSIONS:Our findings confirm the safety of leukocyte reduction as a strategy in preventing TT-CMV in high-risk allogeneic SCT recipients.

Chapter 1. Introduction. Should the CEO be Rewarded or Punished for Events Beyond the CEO's Control? Warren E. Buffett on Executive Compensation. CEO Compensation Theories. CEO Forced Exit Packages. Private Equity Compensation. Entertainment and Sports Celebrities. Benefits of Good Corporate Governance. Chapter 2. Motivating Executive Performance. Tying Performance to the Strategic Plan. Unintended Consequences. Firm Expansion and CEO Pay. Equity-Based Compensation. A New Role for CEOs. Minimum Equity Ownership Requirements. Chapter 3. Peer Groups and Benchmarking. Adjusting the Peer Group. Peer Groups for Different Levels of Executives. Benchmarking. Lake Wobegon Effect. Benchmarking with Medians. Chapter 4. Competing with Private Equity Funds. Chapter 5. Explaining Eexcutive Compensation to Shareholders. Introduction. Reconstructing Executive Compensation Disclosure for Shareholders. Satisfying Investor Expectations. Chapter 6: Compensation Committee Ordinary Operations. SEC Action against Tyson Foods. Earnings on Deferred Compensation. Practical Steps for Compensation Committees. Other Best Practices. Chapter 7. Negotiating Executive Employment and Severance Agreements. Negotiating With New CEO Candidates. Negotiating With Existing CEOs. Best Practices Applicable to New and Existing CEOs. Council of Institutional Investors. Gross-Up Clauses. Retirement Arrangements. The Grasso Case. Section 409A of the Internal Revenue Code. Best Practices in Negotiating Employment or Severance Agreements. Chapter 8. Compensation Committe Structure and Process. Creating Incentives for Good Corporate Governance. Disney Litigation. Chapter 9. Equity Incentive Choices. Overview of Equity Incentives for Key Employees. Dilution. Stock Option versus Stock Appreciation Rights. Restricted Stock versus SARs or Phantom Appreciation Plans. Phantom Plans. ISOs versus Non-ISOs. The Advantage of ISOs. Non-ISOs with Tax Reimbursement. The Tax Benefit to the Company. Chapter 10. Option Granting Practices. Option Granting Practices. The Council of Institutional Investors. Equity Retention Practices. Chapter 11. Director Compensation. Retainer and Differential Pay. Minimum Equity Requirements. Director Compensation Procedure and Process. Shareholder Approval. Perquisites, Repricing and Exchange Programs, Change in Control and Severance Payments. Disgorgement. Chapter 12. Negotiating for the Executive. CEO Turnover. New Candidates for CEO or Other Executive Positions. Employment Agreements with Private Equity Buyers of CEO's Business. Chapter 13. Executive Compensation and Section 409A of the Internal Revenue Code. Background. Plans That Do Not Provide For the Deferral of Compensation. Nonqualified Deferred Compensation Plan - Plans That Provide For Deferred Compensation. Special Rule Applicable to Specified Employees. Change-in-Control Events. Change in the Ownership of a Corporation. Change in the Effective Control of a Corporation. Change in Ownership of a Substantial Portion of a Corporation's Assets. Structuring the Payments upon an Event Payment Trigger. Specified Time or Fixed Schedule. Equity-Based Compensation. Section 409A Compliance. Appendix A. Compensation Committee Charter. Appendix B. Corporate and Securities Update: SEC Adopts Sweeping Overhaul of its Executive Compensation Disclosure Requirements (September 2006). Appendix C. Employment Agreement. Appendix D. Public Company Equity Incentive Plan. Appendix E. Search Terms Typically Required to be Researched by Public Company in Option Backdating Investigations by the Securities and Exchange Commission. Appendix F. What an Employee Should Know about His or Her Stock Options.

A novel class of Hsp90 inhibitors, structurally distinct from previously reported scaffolds, was developed from rational design and optimization of a compound library screen hit. These aminoquinazoline derivatives, represented by compound 15 (SNX-6833) or 1-(2-amino-4-methylquinazolin-7-yl)-3,6,6-trimethyl-6,7-dihydro-1H-indol-4(5H)-one, selectively bind to Hsp90 and inhibit its cellular activities at concentrations as low as single digit nanomolar.

In the course of our Heat Shock 90 program, certain carbazole compounds were identified which had an off-target antiproliferative activity. To understand the off-target activity, we studied one analog with strong activity. We discovered that it had an effect on tubulin polymerization kinetics and was competitive with colchicine. Additional analogs were made, and a number of potent compounds were identified.

A novel class of heat shock protein 90 (Hsp90) inhibitors was developed from an unbiased screen to identify protein targets for a diverse compound library. These indol-4-one and indazol-4-one derived 2-aminobenzamides showed strong binding affinity to Hsp90, and optimized analogues exhibited nanomolar antiproliferative activity across multiple cancer cell lines. Heat shock protein 70 (Hsp70) induction and specific client protein degradation in cells on treatment with the inhibitors supported Hsp90 inhibition as the mechanism of action. Computational chemistry and X-ray crystallographic analysis of selected member compounds clearly defined the protein-inhibitor interaction and assisted the design of analogues. 4-[6,6-Dimethyl-4-oxo-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]-2-[(trans-4-hydroxycyclohexyl)amino]benzamide (SNX-2112, 9) was identified as highly selective and potent (IC(50) Her2 = 11 nM, HT-29 = 3 nM); its prodrug amino-acetic acid 4-[2-carbamoyl-5-(6,6-dimethyl-4-oxo-3-trifluoromethyl-4,5,6,7-tetrahydro-indazol-1-yl)-phenylamino]-cyclohexyl ester methanesulfonate (SNX-5422, 10) was orally bioavailable and efficacious in a broad range of xenograft tumor models (e.g. 67% growth delay in a HT-29 model) and is now in multiple phase I clinical trials.

Objective. To evaluate the ability of SNX-7081, a novel small molecule inhibitor of Hsp90, to block components of inflammation, including cytokine production, protein kinase activity, and angiogenic signaling. A close analog was evaluated in preclinical in vivo models of rheumatoid arthritis (RA).Methods. SNX-7081 binding to Hsp90 was characterized in Jurkat cells and RA synovial fibroblasts (RASFs). Inhibition of NF-kappa B nuclear translocation was evaluated in cellular systems, using lipopolysaccharide (LPS), tumor necrosis factor alpha, or interleukin-1 beta stimulation. Suppression of cytokine production in THP-1. cells, human umbilical vein endothelial cells, and RASFs was studied. Disruption of MAPK signaling cascades by SNX-7081 following growth factor stimulation was assessed. SNX-7081 was tested in 2 relevant angiogenesis assays: platelet-derived growth factor activation of fibroblasts and LPS-induced nitric oxide (NO) release in J774 macrophages. A close analog, SNX-4414, was evaluated in rat collagen-induced arthritis and adjuvant-induced arthritis, following oral treatment.Results. SNX-7081. showed strong binding affinity to Hsp90 and expected induction of Hsp70. NF-kappa B nuclear translocation was blocked by SNX-7081 at nanomolar concentrations, and cytokine production was potently inhibited. Growth factor activation of ERK and JNK signaling was significantly reduced by SNX-7081. NO production was also sharply inhibited. In animal models, SNX-4414 fully inhibited paw swelling and improved body weight. Scores for inflammation, pannus formation, cartilage damage, and bone resorption returned to normal.Conclusion. The present results demonstrate that a small molecule Hsp90 inhibitor can impact inflammatory disease processes. The strong in vivo efficacy observed with SNX-4414 provides preclinical validation for consideration of Hsp90 inhibitors in the treatment of RA.

3943 As the molecular chaperone Hsp90 has emerged as a promising therapeutic target for the treatment of cancer, attention has increasingly focused on the biochemical mechanism of its inhibition. Most of this attention has focused on the apparent inconsistency between the low micromolar in vitro Hsp90 binding potency of geldanamycin and its analogs compared to their low nanomolar activity against cultured cancer cells. Many hypotheses have been put forward to explain this inconsistency including differences between purified and complexed Hsp90, differences in affinity between normal and tumor Hsp90, reduction of the analogs to their dihydroquinones, differential activity against the 4 Hsp90 family members and unique kinetics of Hsp90 inhibition. We present here further evidence of a time dependent shift in affinity for several ATP competitive Hsp90 inhibitors. This time-dependent shift in affinity correlates with the anti-proliferative activity of these compounds against cancer cell lines. In particular, we describe the development of a proteomics-based assay that profiles the ability of compounds to competitively bind to the ATP pocket of all Hsp90 isoforms in a time-dependent manner, the first reported assay for all Hsp90’s. This assay was applied to geldanamycin analogs, radicicol analogs, purine analogs and several of Serenex’s novel Hsp90 inhibitors. While geldanamycins and the Serenex inhibitors exhibit profound time-dependent binding, other classes of Hsp90 inhibitors do not. In fact, geldanamycin and SNX-2112 show an 11-fold and 14-fold increase in potency respectively when drug incubation time is increased from 5 minutes to 1.5 hours. In addition, analysis of the extensive SAR set of Serenex small molecule Hsp90 inhibitors as well as geldanamycin and its analogs reveal that the kinetics of binding can discriminate between Hsp90 family members and thereby create selectivity within the Hsp90 axis. This assay has enabled the identification of compounds with optimal selectivity and binding kinetics and demonstrates that these kinetics correlate with increased anti-proliferative potency against cancer cell lines. Indeed, unlike geldanamycin and its analogs, the Serenex clinical molecule, SNX-2112, is highly potent against the alpha and beta isoforms of Hsp90 but lacks significant activity against the others, Grp94 and Trap1. A model for the modulation of binding kinetics based on x-ray co-crystal data is presented.

Hsp90 maintains the conformational stability of multiple proteins implicated in oncogenesis and has emerged as a target for chemotherapy. We report here the discovery of a novel small molecule scaffold that inhibits Hsp90. X-ray data show that the scaffold binds competitively at the ATP site on Hsp90. Cellular proliferation and client assays demonstrate that members of the series are able to inhibit Hsp90 at nanomolar concentrations.

The advent of multiple high-throughput technologies has brought drug discovery round almost full circle, from pharmacological testing of compounds in vivo to engineered molecular target assays and back to integrated phenotypic screens in cells and organisms. In the past, primary screens to identify new pharmacological agents involved administering compounds to an animal and monitoring a pharmacologic endpoint. For example, antihypertensive agents were identified by dosing spontaneously hypertensive rats with compounds and observing whether their blood pressure dropped. In taking this phenomenological approach, scientists were focused on the final goal, in this example lowering of blood pressure, rather than developing an understanding of the target, or targets, the compounds were impacting. With the evolution of rational target-based approaches, scientists were able to study the direct interaction of compounds with their intended targets, expecting that this would lead to more-selective and safer therapeutics. With the industrialization of screening, referred to as HTS, hundreds of thousands of compounds were screened in robot-driven assays against targets of interest (with this goal in mind). However, an unintentional outcome of the migration from in vivo primary screens to highly target-specific HTS assays was a reduction in biological context caused by the separation of the target from other cellular proteins and processes that might impact its function. Recognition of the potential consequences of this over-simplification drove the modification of HTS processes and equipment to be compatible with cellular assays.

L'invention concerne des amides heteroaromatiques (ou un sel pharmaceutiquement acceptable de ces derniers) tels que definis dans le descriptif, des compositions pharmaceutiques contenant ces derniers, et leur utilisation en tant qu'inhibiteur du facteur Xa. L'invention concerne egalement un procede de preparation ainsi que des intermediaires utilises dans la preparation.