This dissertation comprises an introduction, background and current research progress in the area of classical antifolates as the targeted anticancer therapies.<br>In this study, twelve series of classical 5-, 6- and 7-substituted pyrrolo[2,3-d]pyrimidines were designed and synthesized. Extensive structure modifications of the pyrrolo[2,3-d] pyrimidine scaffold were investigated to determine selective transport via FR or/and PCFT and tumor targeted antifolates with GARFTase or multiple folate metabolizing enzyme inhibition.<br>The design strategies employed include: variation of the side chain substitution position (5-,6- and 7-substituted); variation of the side chain length (n=1-6); isosteric replacement of the 1,4-disubstituted phenyl ring with 1,2- and 1,3- disubstituted phenyl ring and 2,5- disubstituted thiophenyl ring; replacement the L-glutamate with variation at the á and ã carboxylic acids.<br>As a part of this study, a total of one hundred and fifty six new compounds (including new intermediates) were synthesized and separated. Of these, twelve series consisting of forty two classical antifolate final compounds were submitted for biological evaluation. In addition, bulk synthesis of some potent final compounds (2, 2.0 g; 161, 500 mg; 175, 1.0 g; 166, 500 mg; 194, 500 mg) was carried out to facilitate in vivo evaluation.<br>More importantly, a new Heck coupling of the thiophene iodide 301 and allyl alcohols to synthesize aldehydes in one step was discovered. Due to its potential use in analog synthesis of clinically used antifolates such as RTX and PMX, this mild conditioned and easy to handle Heck coupling reaction is highly attractive.<br>During this study, the SAR of folate transporters (RFC, FR and PCFT) and GARFTase inhibitors were extensively explored. The 6-substituted straight chain compound 166 (n=7) was extremely potent against KB tumor cells (IC50=1.3 nM, about 80-fold more potent than clinically used PMX) without any RFC activity. The 5- substituted phenyl compound 175 (n=4) showed AICARFTase as the primary target with potent KB tumor cell inhibition (IC50=7.9 nM, about 8-fold more potent than PMX) and also indirectly inhibited the mTOR pathway leading to tumor cell apoptosis. Due to their potent antitumor activities, these two compounds serve as leads for future structural optimization. / Mylan School of Pharmacy and the Graduate School of Pharmaceutical Sciences; / Medicinal Chemistry / PhD; / Dissertation;

Parkinson’s disease (PD) is a common neurodegenerative disorder that is characterized by the degeneration of dopaminergic (DA) neurons in the substantia nigra and motor deficits. Although the majority of PD cases are sporadic, several genetic defects in rare familial cases have been identified. Animal models of these genetic defects have been created and have provided unique insights into the molecular mechanisms of the pathogenesis of PD. However, the etiology of PD is still not well understood. Here, taking advantage of the unique features offered by zebrafish, I characterized the functions of PINK1 (PTEN-induced kinase 1) gene, which is associated with recessive familial PD, in the development and survival of DA neurons. In zebrafish, antisense morpholino knockdown of pink1 did not cause a large loss of DA neurons in the ventral diencephalon (vDC), but the patterning of these neurons and their projections were perturbed. The pink1 morphants also showed impaired response to touch stimuli and reduced swimming behaviour. Moreover, the pink1 knockdown caused a significant reduction in the number of mitochondria, as well as mitochondrial morphological defects such as smaller size or loss of cristae, thus affecting mitochondrial function. These results suggest that zebrafish pink1 plays conserved important roles in the development of DA neurons and in the mitochondrial morphology and function. To better follow DA neurons after injury or administration of toxins, I generated a transgenic zebrafish line, Tg(dat:EGFP), in which the green fluorescent protein (GFP) is expressed under the control of cis-regulatory elements of dopamine transporter (dat). In Tg(dat:EGFP) fish, all major groups of DA neurons are correctly labeled with GFP, especially the ones in the vDC, which are analogous to the ascending midbrain DA neurons in mammals. In addition, we observed that the DA neurons in the vDC could partially be replaced after severe laser cell ablation. This suggests that zebrafish may have the unique capacity of regenerating DA neurons after injury. Taken together, my studies suggested that zebrafish could be a useful alternative animal model for the study of the molecular mechanisms underlying PD and for the screening of potential therapeutic compounds for PD.

The interplay of transporters and enzymes and their transporter-enzyme was examined in Caco-2 cell monolayer and recirculating perfused rat liver preprations via both theoretical and experimental approaches. First, a Caco-2 catenary model that consisted of the apical, cellular, basolateral compartments and encompasses influx, efflux transporters and enzymes was shown to be superior to the single barrier approach for data interpretation on transporter- and enzyme- mediated processes. The kinetics of baicalein, a flavonoid that undergoes glucuronidation and sulfation, were found to be described better by the catenary model for the complex kinetics of substrate inhibition in metabolism. Second, estradiol-17beta-D-glucuronide (E217G), a protypic substrate of Oatp1a1, 1a4, and 1b2 and Mrp2 that underwent futile cycling with its 3-sulfate metabolite (E23S17G) via estrogen sulfotransferase (Sult1e1) and arylsulfatase C, was examined in the perfused rat liver preparation. Solutions of the AUC and clearances were solved to relate the intrinsic clearances of transporters and enzymes to understand how these affected the apparent clearances in the presence of futile cycling. Transporters and enzymes were perturbed experimentally by the intraportal injection of CC531 colon carcinoma cells for tumor induction in Wag/Rij rat livers. The protein expression of Oatp1a1 and Oatp1b2 were reduced to half whereas Sult1e1 was increased by 40% with tumor development versus the sham-operated control. These data were well predicted by the physiologically-based liver model, showing the impact of increased sulfation intrinsic clearance but not the decreased influx clearance. The TR- (Mrp2 mutant) rat model was used to examine how the absence of Mrp2 for biliary secretion of both E217G and E23S17G affected futile cycling. Absence of Mrp2 was found to result in a pseudo steady-state and reduction of the total, excretion, and metabolic clearances in the liver. The work shed new insight on the interplay between enzymes and transporters and how kinetic processes mediated by enzymes or efflux transporters affected futile cycling.

Detta examensarbete är utfört på uppdrag av Vägverket Region Syd. Sveriges vägnät håller på att anpassas till den europeiska standarden, men det finns fortfarande en hel del begränsningar kvar på mindre vägar. Vi har inte tillräcklig kunskap om vilka problem dessa begränsningar medför för tunga transporter och Vägverket är angeläget om att klarlägga, vilken nytta det skulle vara att göra förstärkningsåtgärder. Jag har undersökt ett landsbygdsområde i södra Östergötland och de begränsningar som finns här på flera vägar är dålig bärighet i första hand för broarna och hög tjälfarlighet. Syftet med undersökningen är att göra en kartläggning av de tunga godstransporterna och de problem begränsningarna på vägarna medför för näringslivet. Jag har använt både teoretiska och empiriska studier. De teoretiska studierna kan delas in i: - Allmänna tekniska beskrivningar för vägkonstruktioner - Lagar och bestämmelser för tunga fordon - Information och rapporter från Vägverket - Tidigare utförda undersökningar De empiriska studierna har utgjorts av intervjuer med olika transportörer i området. Intervjuerna har utförts för att få en bild av hur transportörerna uppfattar vägnätets begränsningar och hur detta påverkar deras verksamhet både praktiskt och ekonomiskt. Jag har intervjuat både stora företag som Södra Skog, Arla och Odal och mindre åkeriägare med ett fåtal lastbilar. Mina slutsatser är följande: 1. Det är fyra vägar i det undersökta området som bör prioriteras när det gäller förstärkningsåtgärder och tjälsäkring. 2. Vägverket bör undersöka möjligheterna att säkerställa en lätt och smidig kontakt mellan transportörer och de som ansvarar för underhållet av vägarna. 3. Det verkar vara bättre hålla kvar de mindre grusvägar som finns kvar och underhålla dem bättre med hyvling, grus och eventuellt dikning i stället för att lägga en tunn asfaltbeläggning, som är mycket känslig för tung trafik och tjällossning.

The interplay of transporters and enzymes and their transporter-enzyme was examined in Caco-2 cell monolayer and recirculating perfused rat liver preprations via both theoretical and experimental approaches. First, a Caco-2 catenary model that consisted of the apical, cellular, basolateral compartments and encompasses influx, efflux transporters and enzymes was shown to be superior to the single barrier approach for data interpretation on transporter- and enzyme- mediated processes. The kinetics of baicalein, a flavonoid that undergoes glucuronidation and sulfation, were found to be described better by the catenary model for the complex kinetics of substrate inhibition in metabolism. Second, estradiol-17beta-D-glucuronide (E217G), a protypic substrate of Oatp1a1, 1a4, and 1b2 and Mrp2 that underwent futile cycling with its 3-sulfate metabolite (E23S17G) via estrogen sulfotransferase (Sult1e1) and arylsulfatase C, was examined in the perfused rat liver preparation. Solutions of the AUC and clearances were solved to relate the intrinsic clearances of transporters and enzymes to understand how these affected the apparent clearances in the presence of futile cycling. Transporters and enzymes were perturbed experimentally by the intraportal injection of CC531 colon carcinoma cells for tumor induction in Wag/Rij rat livers. The protein expression of Oatp1a1 and Oatp1b2 were reduced to half whereas Sult1e1 was increased by 40% with tumor development versus the sham-operated control. These data were well predicted by the physiologically-based liver model, showing the impact of increased sulfation intrinsic clearance but not the decreased influx clearance. The TR- (Mrp2 mutant) rat model was used to examine how the absence of Mrp2 for biliary secretion of both E217G and E23S17G affected futile cycling. Absence of Mrp2 was found to result in a pseudo steady-state and reduction of the total, excretion, and metabolic clearances in the liver. The work shed new insight on the interplay between enzymes and transporters and how kinetic processes mediated by enzymes or efflux transporters affected futile cycling.

Parkinson’s disease (PD) is a common neurodegenerative disorder that is characterized by the degeneration of dopaminergic (DA) neurons in the substantia nigra and motor deficits. Although the majority of PD cases are sporadic, several genetic defects in rare familial cases have been identified. Animal models of these genetic defects have been created and have provided unique insights into the molecular mechanisms of the pathogenesis of PD. However, the etiology of PD is still not well understood. Here, taking advantage of the unique features offered by zebrafish, I characterized the functions of PINK1 (PTEN-induced kinase 1) gene, which is associated with recessive familial PD, in the development and survival of DA neurons. In zebrafish, antisense morpholino knockdown of pink1 did not cause a large loss of DA neurons in the ventral diencephalon (vDC), but the patterning of these neurons and their projections were perturbed. The pink1 morphants also showed impaired response to touch stimuli and reduced swimming behaviour. Moreover, the pink1 knockdown caused a significant reduction in the number of mitochondria, as well as mitochondrial morphological defects such as smaller size or loss of cristae, thus affecting mitochondrial function. These results suggest that zebrafish pink1 plays conserved important roles in the development of DA neurons and in the mitochondrial morphology and function. To better follow DA neurons after injury or administration of toxins, I generated a transgenic zebrafish line, Tg(dat:EGFP), in which the green fluorescent protein (GFP) is expressed under the control of cis-regulatory elements of dopamine transporter (dat). In Tg(dat:EGFP) fish, all major groups of DA neurons are correctly labeled with GFP, especially the ones in the vDC, which are analogous to the ascending midbrain DA neurons in mammals. In addition, we observed that the DA neurons in the vDC could partially be replaced after severe laser cell ablation. This suggests that zebrafish may have the unique capacity of regenerating DA neurons after injury. Taken together, my studies suggested that zebrafish could be a useful alternative animal model for the study of the molecular mechanisms underlying PD and for the screening of potential therapeutic compounds for PD.

Introduction Heteromeric amino acid transporters (HATs) are composed of a heavy subunit (rBAT or 4F2hc) and a light subunit (b0 + AT, ASC1, LAT1, LAT2, y + LAT1, y + LAT2 and xCT), joined by a disulfide bridge (Chillaron et al. 2001). rBAT and 4F2hc are type II membrane glycoproteins (N-terminal cytoplasmic). Both have a single transmembrane segment, an N-terminal intracellular tail and an extracellular domain (ectodomain). As far as we know, the role of the heavy subunit is facilitating the transit of the light subunit to the plasma membrane. The light subunits are polytopic proteins unglycosylated, with 12 transmembrane segments, and the N-and C-terminal intracellular. The light subunit is the catalytically active subunit which confers specificity to the heterodimer on the transport system (Reig et al. 2002): LAT1 and LAT2 for system L , y+LAT1 and y+LAT2 for system y + L, asc for system ASC1; xCT for system Xc -, and b0+at for system b0, + (Chillaron et al. 2001). Results Overexpression of these human proteins was carried out with the methylotrophic yeast Pichia pastoris (strain KM71H) as expression system (based on Long et al. 2005). The main objective was to generate enough protein in a high level of purity to study the structure and check their function by transport assays. The different subunits, light and heavy, were cloned into the expression vector pPICZ (Invitrogen). To facilitate the purification of the different proteins, a cluster of 10 histidines was introduced by PCR at the N-terminus of the heavy subunits and a StrepTagII (IBA) at the N-terminus of the light subunits. 4F2hc is a glycoprotein with 4 possible targets for glycosylation. The glycosylations confer heterogeneity to protein, thus glycosylation targets were eliminated by directed mutagenesis. From all these human heavy and light subunits and heterodimers, only 4F2hc for the heavy subunits, LAT2 for the light subunits, and the heterodimer 4F2hc/LAT2 were overexpressed and extracted from the yeast membrane in enough amounts to continue with the purification step. The light subunit LAT2 was successfully purified but when the stability was analysed by size exclusion chromatography showed a clear profile of aggregation, concluding further studies. In contrast, the heavy subunit 4F2hc was stable after the exclusion chromatography for two days. The heterodimer 4F2hc/LAT2 proved to be stable after gel filtration analysis during one day. Thus, the heterodimer was significantly more stable than the light subunit alone, which allowed us to make an important statement. The catalytic subunit LAT2 needed their heavy subunit (i.e. 4F2hc) to increase the stability. This statement contrasted with the results for the heterodimer rBAT/b0+AT, in which was the heavy subunit rBAT the one who needed its light subunit b0+AT to a correct folding during its biogenesis (Bartoccioni et al. 2008; Rius et al. 2011). Functional studies with human heterodimer 4F2hc/LAT2 were set up to check the role of 4F2hc in the transport. Firstly the functionality of the heterodimer 4F2hc/LAT2 and the light subunit LAT2 in the living cell was checked successfully, meaning a correct folding at expression level. The apparent KM in both cases was the same, remaining unanswered the role of the heavy subunit 4F2hc in the transport function. Next, reconstitution in liposomes was carried out successfully for 4F2hc/LAT2 but not for LAT2, due to the high aggregation tendency. 4F2hc/LAT2 showed the typical overshoot for an amino acid transporter. To carry out the structural studies and due to the difficulty to maintain a stable soluble heterodimer, it was decided to carry out the technique of Single particle -negative staining (SP-NS) in the laboratory of Prof. Fotiadis in the University of Bern (Switzerland). The 3D model technique based on transmission electron microscopy (TEM) is relatively new and has been imposed for mammalian membrane proteins, allowing structural analysis with relatively small concentration of protein. The pure heterodimer was stained in a grid with uranyl formate at 0.75% (two drops optimized for 1 second, washing with water twice). This sample was analysed by transmission electron microscopy (TEM). Different images of projections in different orientations for 4F2hc/LAT2 were kept in a library of 11,000 picks. The refinement of the whole library allowed the 3D reconstruction of this protein by Mr. Meury. The model showed two asymmetric particles, one smaller, in which the crystal of the human ectodomain 4F2hc (Fort et al. 2007) fitted pretty well, and other bigger, which showed a black hole. Thus, the smaller particle was recognized as the heavy subunit, located on top of the light subunit. The resolution was 19 amstrongs, which was in the normal range for this method (from 16 amstrongs to 25 amstrongs). Discussion It was observed that the heavy subunit was located on top of the light subunit LAT2, and not in contact with the cell membrane as was firstly though. The size for the heavy subunit coincided with the existing 3D crystals of the human ectodomain which can fit quite accurately, always assuming the presence of the transmembrane segment in the 3D model. By contrast, the light subunit did not fit with the crystal structure of the prokaryotic homolog AdiC in the APA family (APC superfamily) (Gao et al. 2009) (Kowalczyk et al. 2011) due to the large amount of detergent surrounding this highly hydrophobic subunit in SP-NS method. In spite of that, when the size was compared with AdiC and Stet (a prokaryotic homolog in the LAT family with 30% of homology) studied in the same SP-NS method (Casagrande et al. 2009) the light subunit LAT2 coincided in size with its homologs, demonstrating that the increased volume was due to the detergent effect. Supporting the 4F2hc/LAT2 model, interaction studies with integrins (Feral et al 2005; Feral et al. 2007) and other membrane proteins involved in cell growth (ICAMI; Liu et al. 2003) and / or overexpressed in tumours (CD147/MCT1; Xu et al. 2005) suggest an effect in the transport function through the heavy subunit 4F2hc, which may be explained with an orientation on top of the light subunit and interaction by the external loops. New Evidences: Recently, the 4F2hc/LAT2 heterodimer model in which the heavy subunit is located on top of the light subunit has been corroborated by cross-linking experiments by Miss Helena Alvarez in our laboratory. This fact, allow us to imagine how interactions between both subunits will carry out also when the disulphide bridge is missing. Analyzing the external loops in AdiC atomic structure (the closest paradigm with LATs at present) is found that the external loop 3 and the external loop 4 are the longest (around 25 residues). These loops are even longer in LAT2, which make possible the interaction between both subunits being the separation of 16 amstrongs in the 3D model. Both loops have important roles in the transport cycle based in LeuT fold. The external loop 3 has an important movement in the transition from outward-open conformation to occluded-outward conformation due to the tilt of 40o of the transmembrane 6. The external loop 4, moves down to lid the substrate pathway during the transition from occluded-outward conformation to the occluded-inward conformation. Our new 3D model of a human heteromeric aminoacid transporter offers the opportunity to study new aspects about the role of the heavy subunit in the holotransporter. If the external loops join 4F2hc and LAT2 modulating the transport function in presence of other transmembrane proteins, or if 4F2hc only acts as a bollard of a multiproteic complex, rest to be studied in the future. / Els transportadors heteromèrics d'aminoàcids (HATS) de metazous estan formats per una subunitat pesada (4F2hc o rBAT) (N-glicoproteïna amb 1 segment transmembrana i un gran ectodomini en el seu extrem C-terminal), i una subunitat lleugera (d'entre 10) unides covalentment per un pont disulfur, fent aquests transportadors únics entre els metazous. En humans, 6 subunitats lleugeres es troben formant heterodímers amb 4F2hc (LAT1, LAT2, y+ LAT1, y + LAT2, XCT i asc1) i una (b0, + AT) amb rBAT. Els HATs tenen incidència en la salut, ja que mutacions en qualsevol de les subunitats ocasionen aminoacidúries (cistinúria, lisinúria amb intolerància a proteïnes), són receptors virals o estan sobre expressats en cèl • lules tumorals. El nostre grup va determinar l'estructura de l'ectodomini de 4F2hc a 2.1 Å (Fort J et al. 2007), i recentment ha resolt l'estructura d'un homòleg procariota (AdiC d' E. coli, amb ~17% d´homologia) de les subunitats lleugeres a 3.0 Å de resolució (Kowalczyk et al. 2011). Per contra no hi ha informació estructural sobre els holo-transportadors HAT. El present treball ens mostra el primer model estructural a 19 Å d'un transportador HAT humà, el transportador 4F2hc/LAT2. La importància de 4F2hc, a part de tenir un paper important en immunologia, es troba en la seva sobreexpressió en cèl•lules tumorals, el que la converteix en una important diana per a tractaments i desenvolupament de vacunes contra el càncer. El model ens mostra com en aquest transportador, l´ectodomini de 4F2hc està situat sobre LAT2, suggerint interacció amb els bucles extracel•lulars del transportador i nos sols interacció a través del pont disulfur del segment transmembrana com es pensava anteriorment. Aquesta nova topologia explica la necessitat i la importància de que l'ectodomini de 4F2hc formi part de l´heterodímer 4F2hc/LAT2 i presenta un escenari estructural per al paper "chaperone-like" de 4F2hc sobre les subunitats lleugeres.

Green Cargo driver idag ett antal terminaler i Sverige i syfte att främja transporter på järnväg och ser möjligheter i att utveckla en av sina terminaler i Göteborg, Göteborg Norra, till en så kallad dry port, alltså en intermodal terminal i inlandet. Detta gav upphov till projektet Terminal Göteborg Norra – en möjlig dry port? med målet att undersöka potentialen och omställningarna med att utveckla befintligt kunderbjudande på Göteborg Norra genom att utreda konceptet dry port samt undersöka efterfrågan på marknaden. Syftet med en dry port är att fungera som en förlängning av hamnen och bör enligt forskare erbjuda samma servicetjänster som en hamn, det vill säga förtullning, säker lagring av både lastade och tomma containrar samt underhåll och städning av containrar. En dry port kan ses som ett bra komplement till hamnar som ligger inne i städer och har problem med begränsade utrymmen i hamnområdet och svårt att expandera då ytan inte längre räcker till. Dessutom kan man minska utsläppen av CO2 genom att få mer gods transporterat på järnvägen. Att utveckla Göteborg Norra, som Green Cargo idag driver i egen regi, skulle innebära att befintliga tjänster behöver kompletteras med de tjänster som fullbordar en dry port. Projektet resulterade i en utredning med potentiella intäkter och kostnader som förväntas uppstå vid ett utvecklande av Göteborg Norra till en dry port. Det visade sig att kostnaderna med att utveckla Göteborg Norra till en dry port är fullt möjliga att tjäna in inom den bestämda tidsramen på två år. Trots det så anser projektet att det finns många delar i utvecklandet som talar emot ett fulländat dry port koncept på Göteborg Norra. Då projektet anser att terminalens läge inte är optimalt för en dry port och att det finns begränsade lagringmöjligheter, oklarheter gällande tulltjänster samt att hamnen inte visar något intresse för att utnyttja tjänsten så är det svårt att motivera omställningen till en fulländad dry port. Däremot finns det en del i dry port konceptet som projektet ser positivt till och det skulle innebära att Göteborg Norra tar emot lastade containrar för transport till hamnen. Då projektets marknadsundersökning visade att många speditörer anser att det är problem med långa köer för att hämta och lämna gods i Göteborgs hamn så skulle Green Cargo kunna erbjuda en tjänst där dessa kunder kan åka till Göteborg Norra och lämna eller hämta sitt gods. Detta under förutsättning att Green Cargo kan erbjuda en billigare lösning och att det inte uppstår köer i trafiken runt terminalen. Klarar Green Cargo detta har de ett bra argument som borde locka både speditörer och Göteborgs hamn att nappa på lösningen.

In Streptococcus pyogenes, the protein SiaA (HtsA) is part of a heme uptake pathway system and involved in heme transfer from Shp to the ABC transporter. SiaA mutants, in which alanine replaces the axial histidine (H229) and methionine (M79) ligands, as well as a lysine (K61) and cysteine (C58) located near the heme propionates, are reported. Studies on a mutant of a cysteine expected to be at a distance from the propionates (C47A) are also reported. The coordination state and spin state of the selected mutants were determined via Resonance Raman studies. The pKa values of mutants ranged from 9.0 to 9.4, which were close to the pKa of the WT SiaA (9.7). The midpoint reduction potential of lysine (K61A) mutant was determined by spectroelectrochemical titration to be 61 ± 3 mV vs. SHE, similar to the WT protein (68 ± 3 mV). The addition of guanidinium hydrochloride resulted in protein denaturation that could show more than one process and occurred over days. The ease of protein unfolding was directly related to the extent of interaction of the residues with the heme: changes in the axial ligands resulted in far greater changes in heme protein stability than changes in the residues near the heme propionates. The causative agent of diphtheriae, Corynebacterium diphtheriae, imports heme via an ABC uptake transporter. In this research, two of the five proteins in the heme uptake pathway of C. diphtheriae were studied. These proteins were HmuT, lipoprotein component of the ABC transporter, and HtaA, the heme receptor. UV-visible spectroscopy and fluorescence spectroscopy showed that HmuT protein as isolated bound a porphyrin, rather than heme. Electrospray ionization mass spectrometry (ESI-MS) studies suggested that two tetrapyrroles were bound. To assess stability of this protein towards heme release, thermal denaturation studies were performed. For HtaA, UV-visible and fluorescence spectroscopy also showed the protein as isolated was also bound a porphyrin, rather than heme. Homology studies showed that HtaA protein is quiet distant from homologous heme uptake proteins and could be a member of novel heme binding domain family.

Detta examensarbete har utförts som en del av högskoleingenjörsutbildningen Industriell organisation och ekonomi vid Tekniska Högskolan i Jönköping. Rapporten är skriven på uppdrag av Kinnarps Production AB i Jönköping som är en del av Kinnarpskoncernen. Företaget erbjuder inredningslösningar för kontor och offentliga miljöer och är idag Europas tredje största tillverkare av kontorsmöbler. Produktionen i Jönköping består främst av plåt- och stålbearbetning. Enheten i Jönköping har investerat i en ny monteringslina inom produktionen. Där den nya monteringslinan ska placeras sker i nuläget montering av stolarna Yin och Jig. Därmed måste monteringsplatserna av stolarna flyttas. För närvarande finns ingen given yta att flytta monteringsplatserna till, vilket grundat problemet till detta arbete. Syftet med arbetet är att finna en ny plats för montering av Yin och Jig, samt att effektivisera stolarnas materialflöden. Målet är att presentera ett antal lösningsförslag, innehållande nya monteringsplatser och effektivare materialflöden.  Genom en övergripande kartläggning av produktionsanläggningen i Jönköping och en djupare kartläggning av tillverkningsprocesserna för Yin och Jig, har förståelsen för tillverkningen ökat. Tillgången till egen arbetsplats på företaget har också varit betydande för arbetets resultat. Närheten till produktionen och kontaktpersoner har lett till ett effektivare arbete och en ökad förståelse för Kinnarpskoncernen. En djupare analys av faktorer som är betydande för Yins och Jigs materialflöden har utförts. För att erhålla effektivare materialflöden har faktorerna utvecklats och förbättrats. De faktorer som analyserats är: materialhantering, rörbock, excenterpress, svets, montering och interna transporter. Materialhanteringen är problematisk vid lagring och har förbättrats genom utveckling av lastbärare och lagringsmetod. Rörbocken har identifierats som trång sektor och dess begränsning kan förbättras genom utbildning av personal och införande av säsongsbuffert. Monteringen kommer att utformas med mer flexibla och ergonomiska arbetsplatser. För att minska de interna transporterna kommer materialflödenas ingående operationer att utvecklas mot en flödesgrupp. Detta medför att excenterpressen och robotsvetsen kommer att flyttas till ett gemensamt område med rörlaser och rörbock. Resultatet presenterar tre lösningsförslag som baseras på materialflödesanalysen. Lösningsförslagen innehåller layouter över Yins och Jigs materialflöden och dess nya monteringsplatser. Målsättningarna för de tre layouterna skiljer sig åt. Den första layouten skapar främst ett arbetsteam där operatörer kan samarbeta vid närliggande operationer. För att uppnå detta mål har flera operationer inom Yins och Jigs flöden samordnats. Den andra och tredje layouten skapar främst minskade interna transporter. Detta har uppnåtts genom att skapa rakare materialflöden av Yin och Jig genom produktionsanläggningen. I diskussionskapitlet av lösningsförslagen har layouternas för- och nackdelar tagits upp. I slutsatsen presenteras det starkaste lösningsförslaget.