Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Since its discovery in the 1980’s, HIV has affected the lives of millions of individuals around the globe. Despite obvious need and an enormous amount of research a cure has remained elusive due to the rapid onset of mutated forms of the virus. However, there has been considerable success in reducing viral levels of infected individuals through the use of highly active antiretroviral therapy (HAART). The first-line regimen HAART mainly targets reverse transcriptase (RT) through the employment of two nucleoside RT inhibitors (NRTIs) and a nonnucleoside RT inhibitor (NNRTI). NNRTIs target an allosteric pocket situated about 10 Å from the catalytic site and cause a conformational change in the enzyme upon binding, leading to the inhibition of viral replication. There are currently 5 FDA approved NNRTIs on the market which successfully inhibit viral replication, but the use of these drugs is becoming limited due to the onset of drug resistant strains of the virus. In light of this need for the development of novel NNRTIs, we set out to explore new territory in NNRTI drug design with a goal of maintaining efficacy in the presence of both wild-type and mutated forms of HIV-1. To this end we designed three different NNRTI scaffolds along three different research thrusts. The first of these focused on the synthesis of 15 novel flexible triazole containing compounds. With these compounds we sought to achieve π-π stacking interactions with conserved amino acid residue Trp229 in the hope that we would be able to maintain efficacy in the presence of mutated forms of the virus. An additional feature included hydrogen bonding interactions to the backbone of Lys103. However, despite having thoroughly explored the triazole ring with multiple substitution arrangements, these compounds had very poor to no activity against whole cell HIV-1. Secondly we focused on the synthesis of a 4-hydroxyindole scaffold as a potential NNRTI. The focus here was to achieve interactions to Trp229 and simultaneously achieve hydrogen bonding interactions to the backbone of Lys101 at the entrance of the pocket. This was a novel concept in this class of compounds. We were able to successfully synthesize the indole core as a proofof-concept using the Knoevenagel-Hemetsberger method however; this compound had no activity against HIV-1. Lastly, in our quest to synthesize a novel NNRTI that could maintain efficacy against HIV-1 we decided to attempt to improve upon the stability of a lead indole-based compound synthesized previously within our research group. The lead compound was found to be potent with an IC50 of 1 nM but was unstable in acidic media due to the presence of a methoxy functionality situated at the 3-position on the indole. We sought to overcome this issue by introducing a substituted aryl amine functionality at this position. We were successful in synthesizing our desired compound but unfortunately it was significantly less active against whole cell HIV-1 than the lead compound. However, we were not completely deterred as there are a number of unexplored bioiososteres as possibilities to improve upon the stability of the lead compound while maintaining its excellent activity profile. / AFRIKAANSE OPSOMMING: Sedert die ontdekking van die menslike immuniteitsvirus (MIV) in die 1980’s, het die virus al die lewens van miljoene mense wêreldwyd geaffekteer. Ten spyte van die ooglopende behoefte aan ‘n geneesmiddel sowel as meer navorsing, bly ‘n keermiddel sover onbekombaar as gevolg van die verskillende mutasies wat binne die virus gebeur. Ten spyte hiervan, was daar al heelwat sukses in terme van ‘n verlaging van die virale vlakke in besmette individue deur die gebruik van hoogsaktiewe antiretrovirale terapie (HAART). As ‘n eerste behandeling, teiken HAART meestal trutranskriptase (RT) deur die inspanning van twee nukleosied trutranskriptase inhibeerders (NRTIs) en ‘n nie-nukleosied trutranskriptase inhibeerder (NNRTI). NNRTIs teiken ‘n allosteriese leemte wat ongeveer 10 Å weg van die katalitiese posisie is en veroorsaak dan ‘n konformasie verandering in die ensiem tydens die bindingsproses, wat dan lei tot die inhibisie van die virus se replikasie. Daar is tans 5 FDA goedgekeurde NNRTIs op die mark wat virale replikasie inhibeer, maar die gebruik van hierdie middels word alhoemeer belemmer as gevolg van die onwikkeling van weerstandige stamme van die virus. Met die oog op hierdie nood aan die ontwikkeling van nuwe NNRTIs, het ons gepoog om new gebiede te ondersoek in terme van die ontwerp van NNRTIs, met die doel om die effektiwiteit teen beide die wilde-tipe sowel as die gemuteerde vorme van HIV-1 te behou. Vir hierdie doeleindes het ons drie verskillende NNRTI steiers ontwerp, wat drie navorsingsdoeleindes na streef. Die eerste van hierdie doeleindes was die sintese van 15 nuwe buigsame triasool-bevattende middels. Met hierdie middels het on gepoog om π-π pakkingsinteraksies te behaal met aminosuur residu, Trp229, en sodoende die effektiwiteit van die NNRTIs in die gemuteerde vorm van die virus te behou. ‘n Additionele eienskap wat bygevoeg is, is ‘n waterstofbindingsinteraksie met die ruggraat van Lys103. Ten spyte van pogings om verskeie substitusie patrone om die triasool-ring te ondersoek, het hierdie middels baie swak tot geen aktiwiteit teen heel sel HIV-1 getoon nie. Tweedens, was die fokus op die sintese van ‘n 4-hidroksieindool steier as ‘n potensiele NNRTI. Die fokus hier was om ‘n interaksie met Trp229 te kry terselfdetyd as ‘n waterstofbindingsinteraksie met die ruggraat van Lys101, wat by die opening van die bindingssak is. Hierdie was ‘n nuwe konsep vir hierdie klas van middele. Ons het die indool-kern van hierdie molekules suksesvol gesintetiseer deur middel van ‘n Knoevenagel-Hemetsberger metode, maar ongelukkig het hulle geen aktiwiteit teen HIV-1 getoon nie. Laastens het ons gepoog om ‘n nuwe NNRTI te sintetiseer wat effiktiwiteit teen HIV-1 behou, deur te probeer om vorderings te maak op die stabiliteit van ‘n indool-gebaseerde hoof-middel wat al voorheen deur ons navorsingsgroep geraporteer is. Hierdie hoof-middel het ‘n IC50 waarde van 1 nM gelewer, maar was onstabiel in suur medium as gevolg van die teenwoordigheid van ‘n metoksie-groep in die 3-posisie van die indool. Ons het gepoog om hierdie probleem te oorkom deur ‘n gesubtitueerde arielamien in hierdie posisie te plaas. Ons was suksesvol hierin, maar ongelukkig was die middel heelwat minder aktief teen die heel sel HIV-1 as die metoksie-weergawe. Ten spyte hiervan, is ons optimisties dat ons hierdie probleem kan oorkom, aangesien daar verskeie bioisostere is wat die stabilitiet van middel kan verbeter terwyl dit moontlik die effektiwiteit kan behou.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: With a cure for HIV and AIDS still absent, non-nucleoside reverse transcriptase inhibitors (NNRTIs) play a major role in the current antiretroviral treatments used, which have shown to improve and prolong the lives of HIV patients significantly. However, with rapid mutations of the HI virus, the use of these drugs is becoming limited, thereby highlighting the need for the development of new NNRTIs. Previous work by our research team has led to the development of a cyclopropyl-containing indole-based compound with an inhibition activity (IC50 value) of 0.1 μM, as determined in an in vitro single-cycle, non-replicative phenotypic assay. Therefore, in this project, we focussed on enhancing the intermolecular interactions of our compound to three major areas in the NNRTI binding pocket, namely the Tyr181, the Val179, and the Lys101 binding pockets. Hereby we were able to obtain both improved and lower potencies, with our most active compound having an inhibition activity (IC50 value) of 1 nM. For the interaction to the Tyr181 binding pocket, we were thus unable to synthesise a heterocyclic ring system onto our molecule as opposed to the previously used phenyl ring. Secondly, for the interaction to the Lys101 binding pocket we were able to synthesise a tetrazole ring system and an amide functionality onto the 2-position of the indole. Lastly, in our quest to synthesise the cyclopropyl moiety onto our compound for the interaction in the Val179 binding pocket, we were able to investigate the full inhibition effect of this interaction by synthesising a similar compound with no interaction in this binding pocket. Moreover, we were able to synthesise a new compound with a methoxy moiety for this interaction with an inhibition activity (IC50 value) of 1 nM. With this compound only being submitted for efficacy evaluation as a racemic compound mixture, this opened a new door for research possibilities for our team. / AFRIKAANSE OPSOMMING: In die awesigheid van 'n geneesmiddel vir MIV en VIGS, speel nie-nukleosied omkeerbare transkripsie inhibitore ("NNRTIs")'n groot rol in die huidige antiretrovirale behandeling. Ongelukkig ondergaan die MI virus mutasies, wat dus die gebruik van hierdie antiretrovirale middels beperk. Hierdie beklemtoon dus die noodsaaklikheid vir die ontwikkeling van nuwe "NNRTIs". Vorige werk wat deur ons navorsings groep verrig is, het gelei tot die ontwikkeling van "n siklopropiel bevattende indol verbinding, met "n inhibisie aktiwiteit ("IC50" waarde) van 0.1 μM. Gevolglik, het ons in hierdie projek gefokus om die intermolekulêre interaksies van hierdie verbinding in drie hoof areas in die "NNRTI" bindings ruimte te verbeter, genaamd die Tyr181, die Val179, en die Lys101 bindings ruimtes. Hierdie projek het dus beide verbeterde en ook laer inhibisie aktiwiteits resultate gelewer, waar die mees aktiewe verbinding 'n inhibisie aktiwiteit ("IC50" waarde) van 1 nM behaal het. Vir die interaksie na die Tyr181 bindings ruimte, was ons dus onsuksesvol om 'n heteroaromatiese ring te sintetiseer as plaasvervanger vir die oorspronklike feniel ring. Tweedens, vir die interaksie na die Lys101 bindings ruimte, was ons in staat om 'n tetrazol ring en 'n amied funksionaliteit aan die 2-posisie van die indol te sintetiseer. In ons stryd om die siklopropiel ring aan ons verbinding te sintetiseer vir die interaksie in die Val179 bindings ruimte, was ons in staat om die volledige effek van hierdie interaksie te bepaal deur 'n soortgelykke verbinding te sintetiseer met geen interaksie in die Val179 bindings ruimte nie. Daarenbowe, het ons 'n verbining gesintetiseer met 'n inhibisie aktiwiet ("IC50" waarde) van 1 nM, waarvan die aktiwitiet van slegs die rasemiese mengsel van die verbinding bepaal is. Hierdie vinding het dus 'n nuwe navorsings deur vir ons groep geopen.

published_or_final_version / Pharmacology and Pharmacy / Master / Master of Philosophy

Non-nucleoside reverse transcriptase inhibitor (NNRTI) is one of the key components of antiretroviral drug regimen against human immunodeficiency virus type-1 (HIV-1) replication. However, the low genetic barriers to drug-resistance or cross-resistance, side effects, as well as the unaffordable cost of NNRTIs compromise their clinical usage. Therefore, to develop novel NNRTIs with potent antiviral activity against HIV-1 becomes a major concern in the treatment and prevention of HIV/AIDS. (+)-Calanolide A, which is a natural product initially extracted from the tropical rainforest tree Calophyllum lanigerum, was identified as an attractive NNRTI against HIV-1 despite virus strains containing drug-resistant K103N/Y181C mutations. In this study, a chemical library was constructed based on the three chiral carbon centers of (+)-Calanolide A. After screening the activity against HIVNL4-3 wild-type and several NNRTI-resistant pseudoviruses, a small molecule 10-chloromethyl-11- demethyl-12-oxo-calanolide A (F18) was identified as novel NNRTI with promising anti-HIV efficacy. Further studies were performed to investigate the antiviral breadth, drug resistance profile and underlying mechanism of the action of F18. F18 consistently displayed a potent activity against primary HIV-1 isolates including various subtypes of M group, CRF01_AE, and laboratory-adapted drug-resistant viruses in PBMC based assay. Moreover, F18 displayed distinct profiles against 17 NNRTI-resistant pseudoviruses, with an excellent potency especially against one of the most prevalent strains with the Y181C mutation (EC50=1.0nM) in cell line based assay, which was in stark contrast from the extensively used NNRTIs nevirapine and efavirenz. F18-resistant viruses were induced by in vitro serial passages, and mutation L100I was appeared to be the dominant contributor to F18-resistance, further suggesting a binding motif different from nevirapine and efavirenz. The efficacy of F18 was non-antagonistic when used in combination with other antiretrovirals against both wild-type and drug-resistant viruses in infected PBMCs. Interestingly, F18 displayed a highly synergistic antiviral effect with nevirapine against nevirapine-resistant virus (Y181C). Furthermore, in silico docking analysis suggested that F18 may bind to the HIV-1 reverse transcriptase in a way different to other NNRTIs. For the potential as an anti-HIV-1 microbicide, F18 also showed the stable and rapid release, as well as the sustained antiviral activity against HIV-1 wild-type virus in a formulation temperature-sensitive acidic gel. In summary, this study presents F18 as a new potential drug for clinical use and also underlies new mechanism-based design for future NNRTI. / published_or_final_version / Microbiology / Doctoral / Doctor of Philosophy

Age-related macular degeneration (AMD) is a principal cause of blindness in the United States and other industrialized nations. An estimated 10 million Americans are afflicted with AMD, which is comparable in scope to the 12 million living with cancer, or the 5 million with Alzheimer’s disease. The prevalence of AMD steadily increases with age, affecting 2% of the population at age 40, and one in four people by age 80. For reasons that are not fully understood, AMD is more common in lightly-pigmented and female populations. Treatment of AMD is largely an unmet need: There are no FDA approved therapies except for a small percentage of individuals with end-stage disease. This dissertation investigates the mechanisms of AMD pathogenesis and offers insight into novel therapeutic strategies for this disease.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Beáta Repeľová Supervisor: PharmDr. Lukáš Červený, Ph.D. Title of diploma thesis: Study of effects of antiretroviral drugs on transmembrane transport of tenofovir disoproxil fumarate across MDCKII - ABCB1 cell monolayer Tenofovir disoproxil fumarate (TDF) - ester prodrug of tenofovir is considered as one of the most frequently used component of combination antiretroviral therapy. Several ways of application and good patients' tolerability is typical for this compound. TDF is a substrate of dug transporter such as P-glycoprotein (P-gp) therefore its efflux activity may limit the bioavailability after oral administration and distribution of TDF. As many of antiretroviral drugs are also substrates or inhibitors of P-gp, drug - drug interactions with TDF at the level of transmembrane transport could be expected. The aim of the diploma thesis was to describe effects of co-administered antiretroviral drugs on transfer of TDF across MDCKII cell monolayer by using bidirectional transport and concentration equilibrium setups. The results of experiments confirmed that TDF is a substrate of P-gp. High values of efflux ratio describing transmembrane transport of TDF across parental cells have been observed. This...

Thesis (Ph. D.)--University of Alabama at Birmingham, 2008. / Title from first page of PDF file (viewed Sept. 16, 2008). Includes bibliographical references.

No description available.

by Lam Mei Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 127-137). / Abstract also in Chinese. / Acknowledgment --- p.I / Table of content --- p.II / List of figures --- p.VII / List of tables --- p.IX / Abbreviation --- p.X / Abstract --- p.XII / 論文摘要 --- p.XIII / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Acquired immunodeficiency syndrome --- p.1 / Chapter 1.1.1 --- Discovery of AIDS --- p.1 / Chapter 1.1.2 --- Definition and symptoms of AIDS --- p.1 / Chapter 1.1.3 --- AIDS transmission --- p.2 / Chapter 1.1.4 --- AIDS epidemic --- p.3 / Chapter 1.2 --- Human immunodeficiency virus --- p.3 / Chapter 1.2.1 --- Discovery of HIV --- p.3 / Chapter 1.2.2 --- The structure of HIV --- p.4 / Chapter 1.2.3 --- Genomic structure of HIV --- p.5 / Chapter 1.2.4 --- Life cycle of HIV --- p.5 / Chapter 1.2.5 --- How HIV is involved in different stages of AIDS --- p.7 / Chapter 1.3 --- Therapeutic targets for treatment of AIDS --- p.8 / Chapter 1.3.1 --- HIV reverse transcriptase (HIV RT) --- p.8 / Chapter 1.3.2 --- HIV integrase (HIV IN) --- p.11 / Chapter 1.3.3 --- HIV protease (HIV PR) --- p.12 / Chapter 1.3.4 --- Chemokine receptors --- p.14 / Chapter 1.3.5 --- Vaccine development --- p.16 / Chapter 1.4 --- AIDS therapy --- p.17 / Chapter 1.4.1 --- Current status of AIDS therapy --- p.17 / Chapter 1.4.1.1 --- Drugs approved by US Food & Drug Administration (FDA) --- p.17 / Chapter 1.4.1.2 --- Combination therapy --- p.19 / Chapter 1.4.1.3 --- Vaccine development --- p.19 / Chapter 1.4.2 --- Alternative treatment --- p.20 / Chapter 1.5 --- Objective of my project --- p.21 / Chapter Chapter 2 --- Screening of traditional Chinese medicinal (TCM) plants for HIV reverse transcriptase inhibition --- p.22 / Chapter 2.1 --- Introduction --- p.22 / Chapter 2.1.1 --- HIV RT structure and function --- p.22 / Chapter 2.1.2 --- Natural product against HIV RT --- p.25 / Chapter 2.1.3 --- Inhibitory activities from plant extracts --- p.27 / Chapter 2.2 --- Materials and Methods --- p.28 / Chapter 2.2.1 --- Materials --- p.28 / Chapter 2.2.2 --- Extraction methods --- p.30 / Chapter 2.2.2.1 --- Methanol extraction --- p.30 / Chapter 2.2.2.2 --- Hot water extraction --- p.30 / Chapter 2.2.2.3 --- Preparation of Prunella vulgaris extract --- p.30 / Chapter 2.2.3 --- Reverse transcriptase assay --- p.31 / Chapter 2.2.4 --- Characterization of active component in extract of Prunella vulgaris --- p.32 / Chapter 2.2.4.1 --- Protease digestion --- p.32 / Chapter 2.2.4.2 --- Glucosidase digestion --- p.32 / Chapter 2.2.4.3 --- Ethanol precipitation --- p.33 / Chapter 2.2.4.4 --- Sodium periodiate oxidization --- p.33 / Chapter 2.2.4.5 --- Polyvinylpyrrolidone (PVP) Precipitation --- p.34 / Chapter 2.2.4.6 --- Polyamide resin binding --- p.34 / Chapter 2.2.5 --- Purification of Prunella vulgaris extract --- p.34 / Chapter 2.2.5.1 --- Polyamide resin column chromatography --- p.34 / Chapter 2.2.5.2 --- Sephadex LH-20 chromatography --- p.35 / Chapter 2.2.5.3 --- Reverse phase HPLC chromatography --- p.36 / Chapter 2.2.6 --- Characterization of purified Prunella vulgaris extract --- p.37 / Chapter 2.2.6.1 --- Paper chromatography --- p.37 / Chapter 2.2.6.2 --- Acid hydrolysis of extract --- p.37 / Chapter 2.2.6.3 --- Thin layer chromatography --- p.38 / Chapter 2.2.6.4 --- Other assays --- p.39 / Chapter 2.2.7 --- Calculation --- p.40 / Chapter 2.3 --- Results --- p.41 / Chapter 2.3.1 --- Screening of Herbs --- p.41 / Chapter 2.3.1.1 --- Screening of methanol extracts --- p.41 / Chapter 2.3.1.2 --- Screening of hot water extracts --- p.41 / Chapter 2.3.2 --- Characterization of active components in Prunella vulgaris crude extracts --- p.44 / Chapter 2.3.2.1 --- Protease digestion --- p.44 / Chapter 2.3.2.2 --- Glucosidase digestion --- p.44 / Chapter 2.3.2.3 --- Ethanol precipitation --- p.44 / Chapter 2.3.2.4 --- Sodium periodate oxidation --- p.48 / Chapter 2.3.2.5 --- Effect of naturally occurring chemicals on inhibition of HIV RT --- p.48 / Chapter 2.3.2.6 --- Effect of removal of polyphenolic components of aqueous extract on inhibition of HTV RT --- p.51 / Chapter 2.3.3 --- Further purification of active components in aqueous extract of Prunella vulgaris --- p.53 / Chapter 2.3.3.1 --- Absorption chromatography by polyamide resin --- p.53 / Chapter 2.3.3.2 --- The Sephadex LH-20 chromatography --- p.53 / Chapter 2.3.3.3 --- Reverse phase high performance liquid chromatography --- p.56 / Chapter 2.3.3.4 --- Recovery of extract --- p.59 / Chapter 2.3.3.5 --- Inhibition from extract of various steps of purification --- p.59 / Chapter 2.3.4 --- Characterization of purified aqueous extract of Prunella vulgaris --- p.62 / Chapter 2.3.4.1 --- Paper chromatography --- p.62 / Chapter 2.3.4.2 --- Dose response curve --- p.62 / Chapter 2.3.4.3 --- Acid hydrolysis of purified extract --- p.68 / Chapter 2.3.4.4 --- Identification of monosaccharide in purified extract by Thin layer chromatography (TLC) --- p.71 / Chapter 2.3.5 --- Specificity of the purified extract on polymerase inhibition --- p.75 / Chapter 2.3.5.1 --- Inhibition of purified Prunella vulgaris extract on Taq polymerase --- p.75 / Chapter 2.3.5.2 --- Inhibition of purified Prunella vulgaris extract on Superscript II --- p.75 / Chapter 2.4 --- Discussion --- p.79 / Chapter Chapter 3 --- Screening of inhibitory activities from traditional Chinese medicinal (TCM) plants extracts to HIV protease --- p.86 / Chapter 3.1 --- Introduction --- p.86 / Chapter 3.1.1 --- HIV Protease structure and function --- p.86 / Chapter 3.1.2 --- Natural products against HIV Protease --- p.87 / Chapter 3.1.3 --- Plant extracts against HIV Protease --- p.89 / Chapter 3.2 --- Materials and Methods --- p.91 / Chapter 3.2.1 --- Materials --- p.91 / Chapter 3.2.2 --- Expression of HIV protease --- p.92 / Chapter 3.2.2.1 --- Expression and purification of HIV protease --- p.92 / Chapter 3.2.2.2. --- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.94 / Chapter 3.2.3 --- Characterization of HIV protease --- p.95 / Chapter 3.2.3.1 --- HIV protease assay by fluorometric measurement --- p.95 / Chapter 3.2.3.2 --- HIV protease assay by using reverse phase high performance liquid chromatography --- p.96 / Chapter 3.3 --- Results --- p.98 / Chapter 3.3.1 --- Expression of HIV protease --- p.98 / Chapter 3.3.2 --- HIV protease assay --- p.98 / Chapter 3.3.2.1 --- Protease assay by using reverse phase HPLC --- p.98 / Chapter 3.3.2.2 --- Protease assay by fluorometric measurement --- p.98 / Chapter 3.3.3 --- Screening of crude Chinese medicinal extracts on inhibition of HIV protease --- p.104 / Chapter 3.3.3.1 --- Methanol extracts --- p.104 / Chapter 3.3.3.2 --- Water extracts --- p.105 / Chapter 3.3.4 --- Characterization of herbal extracts on inhibition of HIV protease --- p.110 / Chapter 3.3.4.1 --- Dose response curve of methanol extract of Woodwardia unigemmata --- p.110 / Chapter 3.3.4.2 --- Dose response curve of hot water extract of Prunella vulgaris --- p.110 / Chapter 3.3.4.3 --- Inhibition mode of methanol extract of Woodwardia unigemmata --- p.113 / Chapter 3.3.4.4 --- Inhibition mode of hot water extract of Prunella vulgaris --- p.113 / Chapter 3.3.4.5 --- Effect of partially purified extracts on HIV protease inhibition --- p.116 / Chapter 3.4 --- Discussion --- p.119 / Chapter Chapter 4 --- General discussion --- p.124 / References --- p.127 / Appendix / Appendix 1 Pictures of herbs used in this study --- p.i / Appendix 2 Mass spectrometry of purified Prunella vulgaris extract --- p.vi / Appendix 3 Calibration curve for determination of HIV PR concentration --- p.viii

El virus de la immunodeficiència humana de tipus 1 (VIH-1) és l'agent que provoca la síndrome de la immunodeficiència adquirida (SIDA). Una malaltia, que va ser descoberta al principi del 80, que pateixen avui en dia entre 31.1 i 35.8 milions d'individus arreu del món i que causa 2.7 milions de nous infectats cada any. Per tot això, i pels gairebé tres milions de morts que provoca cada any de mitjana aquesta malaltia, es considera important intentar bloquejar el virus (VIH-1), un paràsit que utilitza la maquinària metabòlica de la cèl·lula per replicar-se. Actualment, el tractament més comú és el TARGA (tractament antiretroviral de gran activitat), que consisteix en la combinació d'inhibidors de la transcriptasa inversa i de la proteasa, dos enzims vírics necessaris perquè es produeixi la replicació del virus. La principal problemàtica que presenta un virus d'aquestes característiques és la gran quantitat de mutacions que pot patir i, en conseqüència, la resistència que adquireix front als fàrmacs administrats. Per això és de vital importància trobar noves molècules que puguin no només suplir-ne d'altres front a aquest problema sinó que puguin interaccionar sobre noves dianes terapèutiques. No va ser fins al segle XXI que van ser aprovats fàrmacs per altres dianes terapèutiques que no fossin la transcriptasa inversa o la proteasa. Destacant-ne Fuzeon® com a inhibidor de fusió entre les membranes del VIH i la cèl·lula hoste i Selzentry® com a inhibidor de CCR5, un dels dos coreceptors d'entrada que pot emprar el virus per infectar la cèl·lula. En canvi, encara a dia d'avui no hi ha cap estructura aprovada per inhibir CXCR4, un altre coreceptor d'entrada. D'entre els compostos estudiats contra aquesta diana terapèutica en destaca l'AMD3100, un compost format per dos macrocicles (ciclam) units per un espaiador p-fenilenbismetilènic. Durant les estudis en fases clíniques, es va observar un efecte cardiotòxic i falta de biodisponibilitat oral relacionats amb l'elevada càrrega positiva a pH fisiològic. En el Laboratori de Síntesi de l'IQS, s'ha desenvolupat una família de tetraamines anàlogues del AMD3100 amb les quals, conservant l'espaiador i substituint les unitats de ciclam per diamines, s'han aconseguit bones activitats inhibidores de CXCR4. En el present treball es desenvolupen variants d'aquestes tetraamines per modificació de l'espaiador i per substitució dels ciclams per altres sistemes heterocíclics, respectivament. D'altra banda, un dels primers ITINANs (Inhibidor de la Transcriptasa Inversa No Anàleg de Nucleòsid) descoberts va ser l'1-[(2-hidroxietoxi)metil]-6-(feniltio)timina (HEPT). En el Laboratori de Disseny Molecular de l'IQS es va seleccionar una quimioteca d'anàlegs d'aquesta estructura i es va proposar una ruta sintètica. Un dels objectius d'aquest treball és doncs, a més d'obtenir anàlegs d'HEPT, confirmar sintèticament la viabilitat de l'itinerari proposat, comprovant que sigui robust, de rendiments elevats i de condicions de reacció prou suaus per poder automatitzar-lo. / El virus de la inmunodeficiencia humana de tipo 1 (VIH-1) es el agente que provoca el síndrome de la inmunodeficiencia adquirida (SIDA). Una enfermedad, que fue descubierta a principios de los 80, que sufren hoy en día entre 31.1 y 35.8 millones de individuos por todo el mundo y que causa 2.7 millones de nuevos infectados cada año. Por todo esto, y por los casi tres millones de muertos que provoca cada año de media esta enfermedad, se considera importante intentar bloquear el virus (VIH-1), un parásito que utiliza la maquinaria metabólica de la célula para replicarse. Actualmente, el tratamiento más común es el TARGA (tratamiento antirretroviral de gran actividad), que consiste en la combinación de inhibidores de la transcriptasa inversa y de la proteasa, dos enzimas víricas necesarias para que se produzca la replicación del virus. La principal problemática que presenta un virus de estas características es la gran cantidad de mutaciones que puede sufrir y, en consecuencia, la resistencia que adquiere frente a los fármacos administrados. Por eso es de vital importancia encontrar nuevas moléculas que puedan no sólo suplir a otras frente a este problema sino que puedan interaccionar sobre nuevas dianas terapéuticas. No fue hasta al siglo XXI que fueron aprobados fármacos para otras dianas terapéuticas que no fuesen la transcriptasa inversa o la proteasa. Destacan Fuzeon® como inhibidor de fusión entre membranas del VIH y la célula huésped y Selzentry® como inhibidor de CCR5, uno de los dos correceptores de entrada que puede usar el virus para infectar la célula. En cambio, aún a día de hoy no hay ninguna estructura aprobada para inhibir CXCR4, otro correceptor de entrada. De entre los compuestos estudiados contra esta diana terapéutica destaca el AMD3100, un compuesto formado por dos macrociclos (ciclam) unidos por un espaciador p-fenilenbismetilénico. Durante los estudios en fases clínicas, se observó un efecto cardiotóxico y falta de biodisponibilidad oral relacionados con la elevada carga positiva a pH fisiológico. En el Laboratorio de Síntesis del IQS, se ha desarrollado una familia de tetraaminas análogas al AMD3100 con las cuales, conservando el espaciador y sustituyendo las unidades de ciclamo por diaminas, se han conseguido buenas actividades inhibidoras de CXCR4. En el presente trabajo se desarrollan variantes de estas tetraaminas por modificación del espaciador y por sustitución de los ciclamos por otros sistemas heterocíclicos, respectivamente. Por otro lado, uno de los primeros ITINANs (Inhibidor de la Transcriptasa Inversa No Análogo de Nucleósido) descubiertos fue el 1-[(2-hidroxietoxi)metil]-6-(feniltio)timina (HEPT). En el Laboratorio de Diseño Molecular del IQS se seleccionó una quimioteca de análogos de esta estructura y se propuso una ruta sintética. Uno de los objetivos de este trabajo es pues, además de obtener análogos de HEPT, confirmar sintéticamente la viabilidad del itinerario propuesto, comprobando que sea robusto, de rendimientos elevados y de condiciones de reacción lo suficientemente suaves para poder automatizarlo. / The human immunodeficiency virus type 1 (HIV-1) is the agent which causes acquired immunodeficiency syndrome (AIDS). A disease that was discovered in the early 80s, that nowadays affects between 31.1 and 35.8 million individuals around the world and causes 2.7 million new infections each year. For all this, and for the nearly three million deaths caused each year on average by this disease, it is considered important to try to block the virus (HIV-1), a parasite which uses the cell's metabolic machinery to replicate. Currently, the most common treatment is HAART (highly active antiretroviral therapy), which is a combination of reverse transcriptase and protease inhibitors, two viral enzymes necessary for virus replication. The main issues presented by this type of virus are the large number of mutations that can occur and, therefore, the resistance it acquires to administered drugs. It is therefore of vital importance to find new molecules which can not only replace others dealing with this problem, but can interact on new therapeutic targets. It was not until the 21st century that drugs interacting with other therapeutic targets, neither reverse transcriptase nor protease, were approved. Emphasizing Fuzeon® as an inhibitor of membrane fusion between HIV and the host cell and Selzentry® as an inhibitor of CCR5, one of the two coreceptor that the virus can use to infect the cell. However, even today there is not an approved structure to inhibit CXCR4, another entry coreceptor. Among the studied compounds against this therapeutic target highlights the AMD3100, a compound formed by two macrocycles (cyclam) linked by a p-phenylenbismethylenic spacer. During clinical trials, there was a cardiotoxic effect and a lack of oral bioavailability associated with its high positive charge at physiological pH. In the Synthesis Laboratory at IQS, a family of tetraamines analogs of AMD3100 which retain the spacer and replace the cyclam units by diamines, have been developed. These compounds showed good CXCR4 inhibitory activity. In this thesis we develop variants of these tetraamines by modification of the spacer and replacement of the cyclams by other heterocyclic systems, respectively. On the other hand, one of the first NNRTIs (Non-Nucleoside Reverse Transcriptase Inhibitors) that was discovered was 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT). In the Laboratory of Molecular Design at IQS a chemical library of analogs of this structure was selected and a synthetic route was proposed. Therefore, one objective of this work is to obtain HEPT analogs, confirm the feasibility of the proposed synthetically pathway, checking the robustness of the reactions, that the yields are high and that the reaction conditions are mild enough to automate the process.