Spandan Chennamadhavuni

SARS-CoV-2 is a positive-sense RNA virus that requires an RNA-dependent RNA polymerase (RdRp) for replication of its viral genome. Nucleoside analogs such as Remdesivir and β-d-N⁴-hydroxycytidine are antiviral candidates and may function as chain terminators or induce viral mutations, thus impairing RdRp function. Recently disclosed Cryo-EM structures of apo, RNA-bound, and inhibitor-bound SARS-CoV-2 RdRp provided insight into the inhibitor-bound structure by capturing the enzyme with its reaction product: Remdesivir covalently bound to the RNA primer strand. To gain a structural understanding of the binding of this and several other nucleoside analogs in the precatalytic state, molecular models were developed that predict the noncovalent interactions to a complex of SARS-CoV-2 RdRp, RNA, and catalytic metal cations. MM-GBSA evaluation of these interactions is consistent with resistance-conferring mutations and existing structure–activity relationship (SAR) data. Therefore, this approach may yield insights into antiviral mechanisms and guide the development of experimental drugs for COVID-19 treatment.

Apurinic/apyrimidinic endonuclease 1 (APE1) is an essential base excision repair enzyme that is upregulated in a number of cancers, contributes to resistance of tumors treated with DNA-alkylating or -oxidizing agents, and has recently been identified as an important therapeutic target. In this work, we identified hot spots for binding of small organic molecules experimentally in high resolution crystal structures of APE1 and computationally through the use of FTMAP analysis (http://ftmap.bu.edu/). Guided by these hot spots, a library of drug-like macrocycles was docked and then screened for inhibition of APE1 endonuclease activity. In an iterative process, hot spot-guided docking, characterization of inhibition of APE1 endonuclease, and cytotoxicity of cancer cells were used to design next generation macrocycles. To assess target selectivity in cells, selected macrocycles were analyzed for modulation of DNA damage. Taken together, our studies suggest that macrocycles represent a promising class of compounds for the inhibition of APE1 in cancer cells.

A modular and diastereodivergent synthesis of tetrahydro-1H-pyrrolo[1,2d]diazepine-(2,5)-diones is presented. The tetrahydropyrrolodiazepinedione scaffold is obtained via a base-mediated three-step isomerization/tandem cyclization of amino acid-coupled homoallylic amino esters. Diastereoselectivity of the process is mediated by the interplay of a kinetic cyclization event and a propensity for thermodynamic epimerization at two labile chiral centers, giving rise to two distinct major diastereomers dependent on starting material stereochemistry and reaction conditions selected. Herein, we present a synthetic and computational study for this tandem process on a variety of amino ester substrates.

// <![CDATA[ $('.header-date').hide();$('#titleAuthors').hide();$('#abstractHeader').hide(); // ]]> Mamoru Shoji1, Wei Ping Qian2, Ganji Purnachandra Nagaraju1, Daniel J. Brat1, 3, Danielle Pessolano4, Rick Luzietti4, Spandan Chennamadhavuni5, 6, Masayoshi Yamaguchi1, Lily Yang2 and Dennis C. Liotta5, 6 1Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University, Atlanta, GA 30322, USA 2Department of Surgery, Emory University, Atlanta, GA 30322, USA 3Department of Pathology, Northwestern University Feinberg School of Medicine and Northwestern Memorial Healthcare, Chicago, IL 60611, USA 4Agilux Laboratories, Inc./Charles River Laboratories, Inc., Worcester, MA 01608, USA 5Emory Institute for Drug Development, Emory University, Atlanta, GA 30322, USA 6Department of Chemistry, Emory University, Atlanta, GA 30322, USA Correspondence to: Mamoru Shoji, email: mshoji@emory.edu Keywords: MACs; UBS109; breast cancer; lung metastases; bone metastasis Received: June 06, 2018 Accepted: September 21, 2018 Published: November 16, 2018 ABSTRACT Synthetic monocarbonyl analogs of curcumin (MACs) are cytotoxic against several cancers including head and neck cancer, pancreatic cancer, colon cancer, and breast cancer. Mechanisms of action include depolarization of the mitochondrial membrane potential and inhibition of NF-κB, leading to apoptosis. We previously demonstrated that UBS109 (MAC), has preventive effects on bone loss induced by breast cancer cell lines. We determined whether UBS109 could inhibit and prevent lung metastasis, since lung metastasis of breast cancer is a major problem in addition to bone metastasis. A breast cancer lung metastasis (colonization) model was created by injection of breast cancer cells MDA-MB-231 into the tail vein of athymic nude mice, nu/nu. Animals were treated with vehicle or UBS109 at 5 or 15 mg/kg body weight by intraperitoneal injection once daily 5 days a week for 5 weeks. UBS109 at 15 mg/kg significantly inhibited lung metastasis/colonization as demonstrated by reduced lung weight consisting of tumor nodules. The body weight of animals treated with UBS109 15 mg/kg remained the same as in the other groups. UBS109 killed completely (100%) MDA-MB-231 breast cancer cells at 1.25 μM in a cytotoxicity assay in vitro. UBS109 15 mg/kg i.p. showed a maximal blood concentration (C_max) of 432 ± 387 ng/mL at 15 min post injection. This is approximately 1.5 ng/ml in the blood of mice and equals 1.5 μM of UBS109. These in vitro and in vivo results are consistent with each other.

The paracaspase MALT1 plays an essential role in activated B cell–like diffuse large B cell lymphoma (ABC DLBCL) downstream of B cell and TLR pathway genes mutated in these tumors. Although MALT1 is considered a compelling therapeutic target, the development of tractable and specific MALT1 protease inhibitors has thus far been elusive. Here, we developed a target engagement assay that provides a quantitative readout for specific MALT1-inhibitory effects in living cells. This enabled a structure-guided medicinal chemistry effort culminating in the discovery of pharmacologically tractable, irreversible substrate-mimetic compounds that bind the MALT1 active site. We confirmed that MALT1 targeting with compound 3 is effective at suppressing ABC DLBCL cells in vitro and in vivo. We show that a reduction in serum IL-10 levels exquisitely correlates with the drug pharmacokinetics and degree of MALT1 inhibition in vitro and in vivo and could constitute a useful pharmacodynamic biomarker to evaluate these compounds in clinical trials. Compound 3 revealed insights into the biology of MALT1 in ABC DLBCL, such as the role of MALT1 in driving JAK/STAT signaling and suppressing the type I IFN response and MHC class II expression, suggesting that MALT1 inhibition could prime lymphomas for immune recognition by cytotoxic immune cells.

Diffuse Large B-cell lymphoma is the most common subtype of B-cell non Hodgkin Lymphoma (B-NHL) representing 30% of all B-NHL. There are three molecular subtypes: germinal center B cell-like (GCB) DLBCL, activated B cell-like (ABC) DLBCL, and primary mediastinal B cell lymphoma (PMBL). Of those, Activated B-cell like Diffuse Large B-cell Lymphoma (ABC-DLBCL) represents a third of the patients and is characterized by constitutive NF-κB activity due to mutations in various proteins of the B-cell receptor (BCR) as well as Toll-like receptor (TLR) pathways. Mutations in the BCR pathway include: activating mutations of CD79A/B (20% of ABC-DLBCLs) and CARD11 (10%) and deletion or inactivating mutation of A20 (20-30%). In the TLR pathway, MYD88 is mutated in 37% of ABC-DLBCL patients. Given this scenario, numerous therapeutic strategies targeting proteins signaling downstream of the BCR pathway have been proposed for ABC-DLBCL -including inhibitors targeting kinases SYK, BTK, PI3K, PKC, MAPKs and AKT or the protease MALT1. MALT1 inhibition provides advantages to other targets because it is downstream of most of the BCR pathway mutations present in patients, including CARD11. Patients with CARD11 mutations do not respond to BTK inhibitors. However, none of the MALT1 inhibitors reported to date are good candidates for clinical use. Here we report a new class of substrate mimetic MALT1 inhibitors based on Z-VRPR-fmk. Our lead compound, SCM-02-138 displayed nanomolar potency in biochemical and cellular MALT1 protease reporter assays. Crystallography shows covalent binding of the compound to MALT1 active site Cys residue. SCM-02-138 was highly selective for MALT1 and showed over 100-fold differential killing in sensitive vs resistant cell lines. MALT1 inhibition was most effective in CARD11 mutant cells. SCM-02-138 was active in vivo, as assessed by hIL10 inhibition in xenografted models of ABC-DLBCL cell lines. Moreover it was effective against ABC-DLBCL xenografts and PDX DLBCL ex vivo. Combination of SCM-02-138 with other BCR inhibitors revealed PI3K delta inhibition as the most synergistic combination. Combination of SCM-02-138 and CAL-101 potentiated both proliferation inhibition and apoptosis. In summary, we have developed a new class of MALT1 peptidic inhibitor characterized by nanomolar potency, irreversibility and high specificity. This inhibitor will be particularly useful against CARD11 mutant ABC-DLBCL, which are resistant to BTK inhibitors. Citation Format: Lorena Fontan, David Scott, John Hatcher, Qi Qiao, Ilkay Us, Gabriella Casalena, Mariette Bekkers, Ulrike Philippar, Matthew Durant, Spandan Chennamadhavuni, Hao Wu, Nathaniel Gray, Ari Melnick. Substrate-mimetic covalent inhibitor of MALT1 is most effective against CARD11 mutant ABC-DLBCL [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-303. doi:10.1158/1538-7445.AM2017-LB-303

Increased activity of transcription factor NF-κB has been implicated in many B-cell lymphomas. We investigated effects of synthetic compound calafianin monomer (CM101) on biochemical and biological properties of NF-κB. In human 293 cells, CM101 selectively inhibited DNA binding by overexpressed NF-κB subunits REL (human c-Rel) and p65 as compared to NF-κB p50, and inhibition of REL and p65 DNA binding by CM101 required a conserved cysteine residue. CM101 also inhibited DNA binding by REL in human B-lymphoma cell lines, and the sensitivity of several B-lymphoma cell lines to CM101-induced proliferation arrest and apoptosis correlated with levels of cellular and nuclear REL. CM101 treatment induced both phosphorylation and decreased expression of anti-apoptotic protein Bcl-X_L, a REL target gene product, in sensitive B-lymphoma cell lines. Ectopic expression of Bcl-X_L protected SUDHL-2 B-lymphoma cells against CM101-induced apoptosis, and overexpression of a transforming mutant of REL decreased the sensitivity of BJAB B-lymphoma cells to CM101-induced apoptosis. Lipopolysaccharide-induced activation of NF-κB signaling upstream components occurred in RAW264.7 macrophages at CM101 concentrations that blocked NF-κB DNA binding. Direct inhibitors of REL may be useful for treating B-cell lymphomas in which REL is active, and may inhibit B-lymphoma cell growth at doses that do not affect some immune-related responses in normal cells.

Macrocycles have the potential to act on currently undruggable targets, but their discovery thus far has been unsystematic. A physicochemical analysis of all nonredundant co-crystal structures now sets out guidelines for macrocycle development. The potential utility of synthetic macrocycles (MCs) as drugs, particularly against low-druggability targets such as protein-protein interactions, has been widely discussed. There is little information, however, to guide the design of MCs for good target protein–binding activity or bioavailability. To address this knowledge gap, we analyze the binding modes of a representative set of MC–protein complexes. The results, combined with consideration of the physicochemical properties of approved macrocyclic drugs, allow us to propose specific guidelines for the design of synthetic MC libraries with structural and physicochemical features likely to favor strong binding to protein targets as well as good bioavailability. We additionally provide evidence that large, natural product–derived MCs can bind targets that are not druggable by conventional, drug-like compounds, supporting the notion that natural product–inspired synthetic MCs can expand the number of proteins that are druggable by synthetic small molecules.

Different chiral dirhodium(II) catalysts are utilized for the intermolecular cyclopropanation of methyl diazoacetates with styrenes.