O limoneno é um produto natural da classe dos terpenos, encontrado abundantemente em plantas cítricas e relatado na literatura como um composto com atividades farmacológicas interessantes, entre elas antibacteriana, antifúngica, antileishmania, nociceptiva e citotóxica. A presente tese relata a funcionalização do limoneno utilizando a Síntese Orgânica em Fase Sólida e a Síntese Orgânica Clássica (em solução). A partir da síntese orgânica em solução, especialmente através das reações de hidroformilação e hidroaminometilação, obtiveram-se vinte compostos. Os compostos foram testados para várias atividades farmacológicas in vitro, a saber: antibacteriana, antifúngica, anti-tripanossoma e anti-leishmania. Destes, dezessete foram testados para a atividade anti-leishmania in vitro contra formas promastigotas de L. (V.) braziliensis e sete apresentaram atividade superior ao fármaco pentamidina, utilizado como padrão no teste, com valores de IC50 entre 11,5 e 35,6 μM. / Limonene is a natural product from the terpene family, found in great proportions in citrical plants, being reported in the literature as having interesting pharmacological activities, such as antibacterial, antifungical, antileishmanial, nociceptive and citotoxic. This thesis reports the funcionalization of limonene via Solid-Phase Organic Synthesis and classical solution-phase synthesis. Twenty products were obtained from the solution-phase protocols, especially from reactions such as hydroformylation and hydroaminomethylation. The compounds were tested for several pharmacological activities, e.g.: antibacterial, antifungical, anti-tripanossomal and anti-leishmanial. Seventeen of those compounds were tested against in vitro promastigote strains of Leishmania (V.) braziliensis and seven compounds were found to have greater anti-leishmanial activity than pentamidine, the standard drug used in this test, presenting IC50 values ranging from 11,5 to 35,6 μM.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-04-29T14:00:29Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Minha dissertação final para biblioteca.pdf: 6771552 bytes, checksum: cc238895a0629230ce30655a760f5a10 (MD5) / Made available in DSpace on 2016-04-29T14:00:29Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Minha dissertação final para biblioteca.pdf: 6771552 bytes, checksum: cc238895a0629230ce30655a760f5a10 (MD5) Previous issue date: 2015-02-19 / CAPEs / Estima-se que até 2030 o câncer será responsável por até 12 milhões de óbitos. Tendo em vista a necessidade de novos tratamentos para o câncer, este trabalho tem como objetivo sintetizar, caracterizar estruturalmente e avaliar a atividade anticâncer de sete novos derivados tiazacridínicos (LPSF AA-56, LPSF AA-57, LPSF AA-59, LPSF AA-60, LPSF AA-61, LPSF AA-62, LPSF AA-63). Estes derivados provém da hibridização molecular dos núcleos de tiazolidina e de acridina. As sínteses para obtenção destes compostos foram otimizadas para obtenção de um melhor rendimento. A síntese da tiazolidina-2,4-diona (TZD) foi feita por reação de ciclização. Uma posterior N-alquilação da TZD em presença de uma base e da 9-(bromo-metil)acridina, conduziu a formação do intermediário LPSF AA-1A. Os intermediários ésteres de Cope (IPs) foram obtidos a partir de uma reação de condensação Knoevenagel. A última etapa ocorreu por uma reação de adição de Michael através de reação entre a TZD N-alquilada com os ésteres de Cope, formando os derivados tiazacridínicos substituídos. A pureza e comprovação estrutural das moléculas sintetizadas foram obtidas através de cromatografia líquida acoplada a espectrômetro de massa (LC-MS), infravermelho (IV) e ressonância magnética nuclear (RMN) de hidrogênio. Ensaios de citotoxicidade dos derivados sintetizados foram realizados em células leucêmicas, fígado e próstata. Entre os compostos sintetizados, o LPSF AA-57 exibiu a atividade anti-cancerígena mais potente contra as linhagens celulares Jurkat (6,63 ± 2,65 μM), HL-60 (6,51 μM), K562 (3,97 ± 1,36 μM) e o LPSF AA-60 exibiu a atividade anti-cancerígena mais potente contra DU 145 (7,22 ± 3,12 μM). / It is estimated that cancer will be responsible for up to 12 million deaths by 2030. Given the need for new treatments for cancer this work aims to synthesize, characterize structurally and evaluate the anti-cancer activity of seven new thiazacridines derivatives (LPSF AA-56, LPSF AA-57, LPSF AA-59, LPSF AA-60, LPSF AA-61, LPSF AA-62, and LPSF AA-63). These derivatives derived of molecular hybridization from nucleus of thiazolidine and acridine. The syntheses for obtaining these compounds were optimized to obtain the best performance. Synthesis of thiazolidine-2,4-dione (TZD) was carried out by cyclization reaction. A further N-alkylation of the TZD in the presence of a base and 9-(bromomethyl)acridine, leading to formation of intermediate LPSF AA-1A. Intermediate esters of Cope (IPs) were obtained from a Knoevenagel condensation reaction. The last step was by a Michael addition, reaction between the N-alkylated TZD with ester of Cope, forming substituted thiazacridines derivatives. The purity and structural confirmation of the synthesized molecules were obtained from liquid chromatography coupled to mass spectrometry (LC-MS), infrared (IR) and Proton nuclear magnetic resonance (NMR). Assays of cytotoxicity of the synthesized products was conducted in leukemic cells, liver and prostate. Among the synthesized compounds, the LPSF AA-57 exhibited the most potent anti-cancer activity against cell lines Jurkat (6,63 ± 2,65 μM), HL-60 (6,51 μM), K562 (3,97 ± 1,36 μM) and the LPSF AA-60 exhibited the most potent anti-cancer activity against DU 145 (7,22 ± 3,12 μM).

Fragment-based virtual screening is an in-silico method that potentially identifies new startingpoints for drug molecules and provides an inexpensive and fast exploration of the relevantchemical space compared to its experimental counterpart. It focuses on docking small potentialbinding fragments to a binding pocket and is used to design improved binders by growing thefragments or joining fragments using suitable linkers. In this project, a fragment-based virtualscreening was evaluated by docking 21 fragments that are obtained from 4 different drugs. Here,the fragments were evaluated using SP score in place and SP and XP flexible docking methodsand were compared to the results of the two decoy fragment datasets. Three of the investigatedfragments are positioned at the top and docked with the correct poses and pockets when comparedto the corresponding substructure in the crystal structure and thus could be considered a successfulfragment starting points. Out of the two flexible docking methods used, the SP method providedadditional correct poses and pockets than XP in this limited dataset.

Le diabète de type 2 (DT2) est une maladie dégénérative liée à une résistance des tissus à l’insuline et à la mort des cellules β-pancréatiques. Il touche près de 400 millions de personnes dans le monde. Les traitements actuels sont symptomatiques et présentent soit des effets secondaires importants soit peu d’effet sur la réduction de la mortalité ou des complications cardiovasculaires. L’agrégation du peptide amyloïde hIAPP (human Islet Amyloid PolyPeptide) est impliquée dans la destruction des cellules β-pancréatiques. A l’état pathologique, hIAPP adopte une conformation en feuillets β, conduisant à son agrégation en formant des oligomères toxiques responsables de la mort des cellules β-pancréatiques. Bloquer cette agrégation est une voie à explorer pour un traitement étiologique du DT2. Des peptides et des peptidomimétiques de type azapeptide, basés sur les séquences peptidiques clés impliquées dans la structuration de hIAPP agrégé, ont été conçus rationnellement dans le but d’interagir avec le peptide de manière sélective et d’inhiber son agrégation. Le développement de différentes voies de synthèse des azapeptides contenant 2 acides aza-aminés consécutifs a été réalisé. L’activité anti-fibrillation des composés a été évaluée par fluorescence à la Thioflavine-T et par microscopie électronique en transmission. Pour les composés les plus prometteurs, l’activité anti-oligomérisation a été évaluée par électrophorèse capillaire et par spectrométrie de masse à mobilité ionique. Ce travail a ainsi permis de développer des composés peptidomimétiques originaux, présentant une activité inhibitrice prometteuse de l’agrégation du peptide amyloïdogénique hIAPP, impliqué dans le diabète de type 2. / Type II diabetes (T2D) is a degenerative disease linked to insulin resistance and pancreatic β-cells death. Worldwide, approximately 400 million people currently have T2D. Actual treatments are symptomatic and have either significant side effects or little effect on reducing mortality or cardiovascular complications. The aggregation of hIAPP peptide (human Islet Amyloid PolyPeptide) is involved in pancreatic β-cells destruction. At the pathologic state, hIAPP adopt a β-sheet rich conformation, leading to its aggregation forming toxic oligomers responsible of pancreatic β-cells death. Blocking this aggregation is a way to explore for etiological treatment of T2D. Peptides and peptidomimetics of azapeptide type, based on key sequences linked to the structuration of aggregated hIAPP, have been rationally designed, to interact with the peptide in a selective manner, and to inhibit its aggregation. The development of different synthetic routes to prepare azapeptides containing 2 consecutive aza-amino acids, have been performed. The anti-fibrillization activity of the compounds have been evaluated by Thioflavin-T fluorescence and by transmission electronic microscopy. For the most promising compounds, the anti-oligomerization activity has been evaluated by capillary electrophoresis and by Ion mobility spectrometry-mass spectrometry. Thus, this work allowed to develop original peptidomimetic compounds displaying a promising inhibitory activity of hIAPP peptide aggregation, involved in type II diabetes.

<p>Alzheimer’s disease (AD) continues to plague the healthcare community as a serious healthcare crisis. Currently there fails to be an effective FDA approved drug that can treat the underlying mechanisms of the disease. Pathologically the disease is characterized by the accumulation of neurotoxic amyloid-b(Ab) plaques within a diseased patient’s brain. These plaques are widely accepted to be generated by the sequential proteolytic cleavage of amyloid precursor protein (APP) by b-secretase (BACE1, memapsin 2) and g-secretase. Numerous biochemical markers suggest that BACE1 is a viable target for AD drug development. Since the cloning and expression of BACE1 there has been an explosion of drug development efforts that have consisted of peptidomimetic-based and non-peptide-based inhibitors. These efforts have led to 13 BACE1 drug candidates some of which have made it to advanced stages of clinical trials. Unfortunately, an effective and tolerable BACE1 drug candidate continues to be rather elusive to the medicinal chemistry community. GRL-8234 is a potent BACE1 inhibitor that has been extensively shown to be tolerable during several short-term and long-term <i>in vivo </i>studies. The scaffold of GRL-8234 provides a suitable template for further lead development. Namely the P1’ 3-methoxy-benzylamine is of particular interest due to the advantageous interactions in the flap region of BACE1 that can be achieved by incorporation of substitution at the benzylic position. In the same way, the <i>meta</i>-substituent of the benzylamine P1’ ligand was investigated to probe the hydrophobic interactions in the S1’-S2’ binding pocket. A novel class of BACE1 inhibitors containing P1’ spirocyclic benzylamine derivatives were designed, synthesized and evaluated for their inhibitory activity against BACE1. In the process of preparing the aforementioned BACE1 inhibitors a method was established to be able to incorporate the desired 3,5-difluorophenyl methyl transition state isostere by utilizing an ester-derived Ti-enolate to access optically pure <i>syn</i>- and <i>anti</i>-aldol adducts as a key step. Additionally, a novel stereoselective method was established to afford heterospirocyclic benzylamines by utilizing a diastereoselective allyl grignard addition to optically pure a-silyloxy <i>N</i>-sulfinyl ketimines as a key step. Biological evaluation of these novel BACE1 inhibitors has been fruitful and continues to be ongoing. </p>

<p>  Acute myeloid leukemia (AML) is a devastating cancer with an overall 5-year survival rate of approximately 30%, despite efforts to develop therapeutics to combat this disease. AML is caused by various mutations, and frequent genetic errors found in 30% of AML patients are mutations in the FMS-like tyrosine kinase 3 (FLT3). Recently two FLT3 drugs for AML, Midostaurin and Gilteritinib, have been approved by the FDA, but resistance to these drugs, such as FLT3D835Y/V and F691L, mutations have limited the efficacies of both of these drugs. Thus, there is a need for newer generation FLT3 inhibitors that cover mutations encountered in the clinic. The Sintim laboratory has developed a FLT3 inhibitor, HSN748, which has shown remarkable efficacies against the majority of the FLT3 mutants. Pre-IND studies are now ongoing to support a potential clinical trial of HSN748 for AML treatment that is resistant to the approved therapeutics midostaurin and gilteritinib. The characterization of HSN748 on the proteome and phosphoproteome level was undertaken to provide a more granular view on how this potential AML therapeutic affects key cellular processes. Global proteomic and phosphoproteomic analysis revealed that HSN748 may play a role in cell cycle regulation, spindle formation, leukemic stem cell maintenance, and transcription and confirmed previous in vitro studies that showed inhibition of other kinases relevant to AML treatment, such as AURKB and Raf-1.</p> <p>Recently, there have been efforts to improve the timespan of therapeutic efficiency of FLT3 inhibitors by combining FLT3 inhibitors with other drugs that target other processes essential to AML. Clinical trials (e.g. NCT03735875) are ongoing to evaluate FLT3 inhibitors combined with venetoclax against AML. The second part of this thesis project evaluated the combination of HSN748 analogs with venetoclax and discovered that the combinations synergize to increase apoptotic activity in AML cells harboring FLT3-ITD with secondary mutations D835Y or F691L, which are two clinically important genetic alterations that lead to drug resistance against current FLT3 inhibitors. We determined that the nicotinamide analog, HSL468, synergized with venetoclax with a coefficient of inhibition of 0.23 for the combination with HSL468 at 1.25 nM and venetoclax at 80 nM against an AML cell line that has both the FLT3-ITD and F691L mutations, which are responsible for resistance to most current FLT3-targeted AML therapeutics, including the combination therapy of gilterinitib and venetoclax.</p>

<p>  </p> <p>Using a version of the Povarov-Doebner reaction, we were able to identify and develop a novel kinase inhibitor scaffold that is tunable, selective, and able to target drug-resistant mutant kinases. The 3H-pyrazolo[4,3-f]quinoline moiety was shown to be a privileged kinase inhibitor scaffold with a strong inhibition several different kinases. Herein, various 3H-pyrazolo[4,3-f]quinoline-containing compounds were synthesized quickly via the Povarov-Doebner multicomponent reaction. Our scaffold has demonstrated to potently inhibit FLT3 and CDK2 with nanomolar IC50 values. These FLT3 inhibitors were also shown to inhibit leukemic cell growth in a mouse disseminated AML model, establishing these 3H-pyrazolo[4,3-f]quinoline-containing compounds as lead compounds to develop into anti-cancer agents. The 3H-pyrazolo[4,3-f]quinoline moiety has potently inhibited ROCK1/2 with single digit nanomolar IC50 values, newly synthesized analogs with replacement of the boronic acid moiety with an amide also displayed inhibition of ROCK1/2. The most active compound (<strong>HSH3107</strong>) potently inhibits ROCK1/2 and although it did not display any antiproliferative effects against MDA-MB-231 (triple negative breast cancer cell line) at 1 µM, it did slow cell migration for up to 48 hours compared to DMSO control and Fasudil. Acyclic amide analogs of this scaffold have also led to the discovery of CDK12 and CDK13 inhibitors which can serve as a potential therapeutic in cancers where there may be no treatment strategy available or where resistance has emerged.</p>

<p>  </p> <p>A concise and scalable total synthesis of the therapeutically relevant methyl ester of the prostaglandin D₂ metabolite, tricyclic-PGDM, was accomplished in 8 steps from a known and easily accessed cyclopentene-diol derivative. The route features three key transition metal catalyzed steps. These steps include: a nickel catalyzed Ueno-Stork type dicarbofunctionalization which generates two consecutive stereocenters on the central cyclopentane core, a late-stage palladium-catalyzed carbonylative oxaspirolactonization, and a <em>Z</em>-selective cross metathesis to introduce the <em>Z</em>-butenoate side chain- a motif difficult to introduce through traditional protocols and which caused significant issues in the previous total syntheses of tricyclic-PGDM. Through this route, we have accumulated 75 mg of material for an ¹⁸O tricyclic-PGDM clinical assay which previously suffered from a material shortage. In addition to completing the synthesis, we generalized the <em>Z</em>-selective cross metathesis and nickel catalyzed Ueno-Stork protocols to numerous other substrates further demonstrating the utility of these transformations. </p> <p><br></p> <p>Protein arginine methyltransferases (PRMTs) catalyze the transfer of methyl groups from the cofactor SAM to arginine residues on various cytosolic and nuclear proteins. Of the nine members of the PRMT family, PRMT5 has been the most extensively studied and has been shown to regulate processes such as the DNA damage response, cell proliferation, and mRNA translation. Although numerous pathways have been identified that regulate PRMT5 activity, the cytosolic protein MEP50 has been identified as a key regulator in many diseases. PRMT5 and MEP50 interact to form a hetero-octameric complex, which can modulate the activity of PRMT5 for many cellular processes. Two new generations of PRMT5:MEP50 inhibitors were strategically designed and synthesized, which do not suffer from chemical instability like our previously most potent analogues. Our best compounds have IC₅₀ values ranging from 512 to 2.5 nM in LNCaP cells, and were confirmed to target the PRMT5:MEP50 interaction through BiFC analysis.</p>

Thesis advisor: Charles Hoffman / The second messenger cyclic 3’-5’ adenosine monophosphate (cAMP) signaling pathway plays an important physiological role in many organisms. Cyclic nucleotide phosphodiesterases (PDEs) regulate signal transduction by catalyzing the hydrolysis of cAMP and cGMP allowing for the downregulation of cyclic nucleotide levels. Human PDEs are encoded by 21 genes grouped into 11 families. The biological role of the most recently discovered PDE family (PDE11) remains poorly understood partly due to the lack of selective inhibitors. Mutations in the PDE11A gene have been linked to a wide range of diseases, such as Cushing Syndrome, which is a result of inactivating mutations expressed in adrenocortical tumors. Meanwhile, PDE11 levels are seen to increase in the ventral hippocampus as a function of aging, and is associated with a loss of social memory. Thus, the development of a selective PDE11 inhibitor could provide a potential therapeutic benefit to patients receiving long-term corticosteroid treatment by stimulating cortisol production by the adrenal gland, as well as to aging adults to maintain social memory. To address these needs, candidate PDE11 inhibitors related to a compound discovered by the Hoffman lab in a high throughput screen for PDE11 inhibitors are being synthesized by the Rotella laboratory. I have been evaluating these compounds using two fission yeast-based growth assays in complement with in vitro enzyme assays carried out by Dr. Jeremy Eberhard. Here I describe my role in the project, leading to the identification of a compound, SMQ2-57, which is a selective inhibitor of the PDE11 enzyme whose potency has been confirmed through both yeast-based assays and in vitro enzyme assays. In addition, I have taken both a forward and reverse genetic approach to identify PDE11A4 mutant alleles that confer resistance to inhibitor compounds as such knowledge could guide a rational drug design approach to produce more effective PDE11 inhibitors. Based on our results, SMQ2-57 could serve as a useful tool in understanding the biological role of PDE11. Meanwhile, data from my study of compound resistant mutant PDE11 alleles should allow for the characterization of the physical interaction between PDE11 and its inhibitors in an effort to guide a medicinal chemistry program to develop a more potent and drug-like PDE11 inhibitor. / Thesis (BS) — Boston College, 2023. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: Scholar of the College. / Discipline: Biology.

The mammalian polyamine transport system (PTS) has been of interest due to its roles in cancer and maintaining cellular homeostasis. Polyamines are essential growth factors which are tightly controlled via a balance of biosynthesis, metabolism, import, and export. This work focused on the development and biological testing of polyamine transport probes to help understand the molecular requirements of the PTS. This was mediated through the use of a CHO (PTS active) and CHO-MG* (PTS deficient) screen, where compounds demonstrating high toxicity in CHO and low toxicity in CHO-MG* were considered PTS selective. The first chapter focused on the development of polyamine-based drugs which are both metabolically stable to polyamine oxidase (PAO) activity and are hyperselective for targeting the PTS. This approach was optimized by combining a di-substituted aryl design with terminal N-methylation of the appended polyamine chains to generate a new class of superior PTS agonists. The metabolic stability of these compounds was demonstrated in CHO and CHO-MG* in the presence and absence of a known PAO inhibitor, aminoguanidine (AG). Highly PTS selective compounds were then tested in the NCI-60 cell line screen to demonstrate the effectiveness of polyamine-based drugs in cancer therapy. During this screen, the MALME-3M (human melanoma) cell line was identified as being very sensitive to these PTS targeting drugs. Further studies using MALME-3M and its normal counterpart, MALME-3, showed excellent targeting of the cancer line over MALME-3. For example, The MeN44Nap44NMe compound showed 59-fold higher toxicity in MALME-3M over MALME-3. The second chapter focused on the development of potential polyamine transport inhibitors (PTIs) for use in combination therapy with ?-difluoromethylornithine (DFMO). This therapy is predicated upon reducing sustained polyamine depletion within cells by inhibiting both polyamine biosynthesis with DFMO and polyamine transport with the PTI ligand. Potential PTIs were identified by blocking the uptake of spermidine in DFMO-treated CHO and L3.6pl cells. Previous work has identified a tri-substituted polyamine-based design as an effective PTI. Low toxicity and a low Ki value in a L1210 screen were good predictors for PTI efficacy. The structural requirements for a potent PTI were explored by modulating the toxicity through the introduction of amide bonds, and also by determining the number and orientation of the polyamine messages (appended to an aryl core) required for efficient inhibition of polyamine uptake. These experiments showed that a tri-substituted design and a triamine message (homospermidine) appended was optimal for PTI potency. The final chapter focused on the development of Dihydromotuporamine C derivatives as non-toxic anti-metastatic agents. Dihydromotuporamine C demonstrated good anti-invasive properties with tumor cells. Derivatives were made in an effort to reduce the cytotoxicity of the parent and improve the anti-migration potency. The motuporamine derivatives all have a polyamine message (norspermidine or homospermidine) appended to make a macrocycle core, making them prime targets to evaluate as potential PTS ligands in the CHO and CHO-MG* screen. Each compound was also tested in the highly metastatic pancreatic cancer cell line L3.6pl to determine both its IC50 value and maximum tolerated dose (MTD). The anti-migration assay was performed at the lowest MTD obtained (0.6 [micro]M) in order to compare the series at the same non-toxic dose. The results suggested that as the N1-amine center was moved further from the macrocyclic ring, an increased ability to inhibit cell migration and reduced toxicity was observed. These collective findings provide new tools for cell biologists to modulate and target polyamine transport in mammalian cells. Future applications of these technologies include new cancer therapies which are cell-selective and inhibit the spread of tumors.