Detta examensarbete är en del i högskoleingenjörsutbildningen vid linneuniversitetet i Växjö. Arbetet är utfört på Ackurat Industriplast AB i Lammhult. Syftet med arbetet är att ta fram samt utveckla fyra av kravelementen i Production Part Approval Process (PPAP) dokumentationen. Kravelementen vilka ingår inom ramen för detta arbete är flödesschema, process FMEA, styrplan samt kontrollinstruktioner. Ackurat har kunder inom fordonsindustrin, dessa kunder ställer höga krav på kvalitén hos produkterna från sina leverantörer. Denna kvalitetssäkring sker dels genom att leverantören skall följa krav ur ISO/ TS 16949 standarden där PPAP är en del av kvalitetssäkringen. Detta innebär att för varje nytt uppdrag så skall PPAP dokumentation bifogas tillsammans med ett utfallsprov. Ackurat upplever att den dokumentation som finns i dagsläget är bristfällig och i behov av uppdatering. Resultatet är utarbetat genom kartläggningsmetoden ” walk through” där en fallstudie av processer har använts . Informationen om processerna har insamlats genom observationer och intervjuer med personal i processerna.  Denna empiristiska information om Ackurats processer har sedan analyserats samt sammanställts med litteraturstudien i teoridelen. Examensarbetet resulterade i fem olika flödesscheman för att illustrera det verkliga flödet i Ackurats processer. Process FMEA utarbetades och åskådliggör felmöjligheter i processflödet. Genom kartläggningsarbetet sammanställdes en styrplan där alla kontroller i processerna ingår. Slutligen utarbetades kontrollinstruktioner för de kontroller som sammanställts i styrplanen.    Arbetet med kartläggningsarbetet av flödesschemat var en viktig del av genomförandet. Detta berodde på att följande dokument är uppbyggda från det aktuella flödesschemat. Stort fokus har således lagts i utvecklandet samt valideringen av de olika flödesschemana. Vid analys av dessa dokument kom författarna fram till att utformade dokument uppfyller syftet samt målen för arbetet. Denna slutsats framgick genom validering med berörd personal på Ackurat samt handledare. Likaväl som att framtagna dokument kan användas i PPAP dokumentation till Ackurats kunder så kan dokumenten användas till det interna förbättringsarbetet i företagets processer.

ABSTRACTION This thesis is focus on discussion of how automobile fastener manufacturer in Taiwan practice QS9000-PPAP and what is different between domestic and foreign automobile fastener suppliers. Chapter 1 makes a brief introduction in this thesis, including research purpose¡Bresearch motive¡Bresearch method and research structure. Chapter 2 describes QS9000 history and discusses some relational writings. Automobile fastener is getting popular in Taiwan fastener industry, because the unit profit is much better than regular parts. Almost automobile parts suppliers in Taiwan only make OEM parts, and because of this reason, it is no need to process fully QS9000 system, however, it is absolute to practice PPAP before mass production. Customers require PPAP based on protection reason, and PPAP can protect customers and OEM factory from any error. At present, there are only few factories in Taiwan can meet standard PPAP requirements, so Chapter 3 will make questionnaires to 50 automobile factories and distributors in the world. The main purpose of questionnaires is to compare main differences of PPAP processes between domestic factories and foreign suppliers and this will greatly help on domestic suppliers production in the future. Chapter 4 and 5 describes standard PPAP procedure and discusses correct FMEA and Measurement system actions, and there are also some examples in this chapter. Chapter 6 accords to the questionnaires results in chapter 3 and analysis the main problem existed in domestic suppliers. Due to many differences between domestic suppliers and foreign suppliers, this chapter will also discuss how to improve the existent differences and try to build a correct concept in PPAP form them. Chapter 7 will summarize this thesis and make some suggestions to domestic automobile suppliers. I hope this thesis will contribute domestic fastener industry a little, and there will be no any claims from customers in the future. Future researchers can continue focus on how to practice QS9000 system in domestic factories without heavy investment.

This thesis is a dissertation to support the development of new domino pericyclic oxy-Cope/ene/Claisen/Diels-Alder reaction, diversity oriented synthesis of PPAPs scaffold via sequential one pot cascade reaction and ethyl aluminum sesquichloride catalyzed highly hindered Diels-Alder reaction. The first part concentrates on the domino pericyclic oxy-Cope/ene/Claisen/Diels-Alder reaction. As a result of this study, we have developed a general methodology for rapidly constructing complex diterpenes and discovered a thermal oxy-Cope/ene/Claisen/Claisen rearrangement, applied to the synthesis of trans decalin benzofurans. The second part involved the development of an efficient synthetic approach towards bicyclo[3.3.1]nonenone core found in many natural products, via a sequential Diels-Alder/gold(I)-catalyzed 6-endo-dig cyclization and its application to the synthesis of a diversified library of PPAPs. Finally, we have developed an efficient synthetic methodology for the formation of cyclohexene rings bearing quaternary carbon centers via an ethyl aluminum sesquichloride mediated highly hindered Diels-Alder reaction. This method solved an important problem encountered in the synthesis of many natural products including PPAPs. This methodology opened new opportunities in the total synthesis of PPAPs.

This thesis is a dissertation to support the development of new domino pericyclic oxy-Cope/ene/Claisen/Diels-Alder reaction, diversity oriented synthesis of PPAPs scaffold via sequential one pot cascade reaction and ethyl aluminum sesquichloride catalyzed highly hindered Diels-Alder reaction. The first part concentrates on the domino pericyclic oxy-Cope/ene/Claisen/Diels-Alder reaction. As a result of this study, we have developed a general methodology for rapidly constructing complex diterpenes and discovered a thermal oxy-Cope/ene/Claisen/Claisen rearrangement, applied to the synthesis of trans decalin benzofurans. The second part involved the development of an efficient synthetic approach towards bicyclo[3.3.1]nonenone core found in many natural products, via a sequential Diels-Alder/gold(I)-catalyzed 6-endo-dig cyclization and its application to the synthesis of a diversified library of PPAPs. Finally, we have developed an efficient synthetic methodology for the formation of cyclohexene rings bearing quaternary carbon centers via an ethyl aluminum sesquichloride mediated highly hindered Diels-Alder reaction. This method solved an important problem encountered in the synthesis of many natural products including PPAPs. This methodology opened new opportunities in the total synthesis of PPAPs.

In terms of Quality, more specifically, the Quality Automotive, we emphasize that the Approval Process Production Parts (PPAP) is characterized by evaluating a series of analyzes of various aspects of manufacturing process of production. Also, refers to the way of reporting the results the process, proving that the vendor has the ability to meet production with the level of quality required by the customer. Quality in need of resources that show where the ERP TECNICON BUSINESS SUITE integrates all departments and business processes with versatility provided by a modular and customizable structure, providing opportunities for solutions aimed at improving communication between all areas of an organization, so specific and complete. Thus, the research aims to define and implement the necessary PPAP procedures for TECNICON company in order to meet the needs of its customers. The implementation is aimed at monitoring and guidance of TECNICON Business Suite users directed to customers. The purpose of use of the system by the company is to provide the use of PPAP for their customers. Thus, the PPAP is characterized by the fact that the need for bureaucratic procedure when clients (where customers TECNICON, are to be PPAP suppliers to their customers) request the documentation on the flow of information between them to review and approval of the PPAP. The methodology was defined as action research, since the researcher provides the search directives during the study period in order to find solutions to the problem in TECNICON in Horizontina (RS). As main results, the implementation stands out PPAP procedures for the use of customers TECNICON Company, was completed from the following tools: Study Capability, DPAR, FMEA, Flow Chart, R & R, Dimensional Report, RAA, RIAI and Linearity. / No âmbito da Qualidade, mais especificamente da Qualidade Automotiva, destaca-se que o Processo de Aprovação de Peças de Produção (PPAP) caracteriza-se por avaliar uma série de análises de vários aspectos de um processo de manufatura. Além disso, refere-se à maneira de relatar os resultados do processo, provando que o fornecedor tem a capacidade de cumprir a produção com o nível de qualidade exigido pelo cliente. Qualidade que necessita de recursos que a comprovam, onde o ERP TECNICON BUSINESS SUITE integra todos os departamentos. Os processos empresariais, com versatilidade proporcionada por uma estrutura modular e personalizável, oportunizam soluções direcionadas à melhoria na comunicação entre todas as áreas de uma organização de maneira específica e completa. Assim, a pesquisa tem como objetivo definir e implantar os procedimentos necessários do PPAP para a empresa TECNICON com o propósito de atender as necessidades de seus clientes. A implantação está voltada ao monitoramento e à orientação dos usuários do TECNICON BUSINESS SUITE direcionada aos clientes. A finalidade da utilização do sistema pela empresa é de proporcionar o uso do PPAP para seus clientes. Dessa forma, o PPAP se caracteriza pelo fato da necessidade do procedimento burocrático, quando os clientes (onde os clientes da TECNICON, passam a serem fornecedores do PPAP para seus clientes) solicitam a documentação referente ao fluxo de informações entre as mesmas para avaliação e aprovação do PPAP. A metodologia foi definida como pesquisa-ação, uma vez que o pesquisador oferece as diretivas de pesquisa durante todo o período de estudo, no sentido de encontrar as soluções do problema na TECNICON, em Horizontina (RS). Como principais resultados, destaca-se a implantação dos procedimentos PPAP para utilização dos clientes da empresa TECNICON, que foi finalizada a partir das seguintes ferramentas: CEP/Estudo de Capabilidade, DPAR, FMEA, Fluxograma, R&R, Relatório Dimensional, RAA, RIAI e Linearidade.

In the course of this work, compounds of the natural product class of polycyclic polyprenylated acylphloroglucinols (PPAPs) were analysed. They consist of a polycyclic core structure decorated with carbonyl groups and unsaturated sidechains. The modular synthesis platform developed and optimised in the work group, facilitated the creation of a 23 compound PPAP library. This initiated the study of antibacterial properties. Further research into the derivatisation of PPAPs paved the way to a synthetic toolbox for the synthesis of new derivatives. Now a follow-up structure-activity relationship (SAR) study, aided by a novel C acylation strategy developed in the group, expanded the library with 25 new unnatural type B PPAPs. Antibiotic efficacies with minimum inhibitory concentrations (MICs) in the nanomolar range against methicillin-resistant Staphylococcus aureus (MRSA) were observed, identifying new lead compounds with significant therapeutic windows. Transitioning to cancer research, the anticancer activities of PPAPs across leukaemia, glioblastoma (GB), and prostate cancer (PCa) were investigated. A 3 phenylpropanoyl head group turned out to be crucial for proapoptotic efficacies. Optimisation efforts yielded different potent compounds for each cancer type with proapoptotic half maximal inhibitory concentration (IC50) values in the low micromolar and antiproliferative IC50 values in the nanomolar range. Flavonoid-like PPAP scaffolds were explored for enhanced biological activities through the combination of anticancer activities of PPAPs with flavonoids. A one-step synthesis method produced eleven flavonoid-like PPAPs with promising anticancer potential, particularly against GB. They exhibited strong proapoptotic effects with low toxicity on healthy cells. To expand the covalent combination of different active substances, further conjugation strategies were employed for the synthesis of PPAP-containing drug-drug conjugates (DDCs). This resulted in several PPAP–temozolomide- (TMZ) and PPAP–doxorubicin (DOX) conjugates. PPAP–TMZ showed significant anticancer potential against GB, while PPAP–DOX demonstrated substantial antileukemic activity. Challengingly, both DDCs also exhibited significant toxic effects on healthy peripheral blood mononuclear cells (PBMCs). Therefore, targeted drug delivery of PPAPs utilising PPAP hybrid drugs was investigated to enhance the selectivity of PPAP treatments. This involved the successful combination of PPAPs with linkers for antibody-drug conjugates (ADCs). To evaluate the coupling potential of the synthesised linker with monoclonal antibodies, it was reacted with cysteine. The successful reaction showed the potential of future couplings with real antibodies. For PCa specifically, prostate specific membrane antigen (PSMA)-targeting PPAPs were explored. This protein is solely expressed by PCa cells, therefore being an ideal target protein. Employing chemical structures from existing pharmaceuticals led to the design of different PSMA-targeting PPAPS. While in silico screenings indicated extraordinarily high affinity to the PSMA binding site, biological tests revealed very low anticancer activities. As the binding affinity was too strong, the PPAPs were presumably not released in the cell, which could explain the low anticancer efficacies. For this reason, cleavable linker systems were investigated as part of further research. In conclusion, this research significantly advanced the understanding of diverse PPAP derivatives, demonstrating their potential in antibacterial and anticancer applications. The outlined synthesis strategies, SAR studies, and conjugation approaches provide a foundation for future developments. Further optimisations and collaborations are needed to bring PPAPs a step closer to applications as medical treatment option.:Table of Contents Acknowledgements III Table of Contents V List of Abbreviations IX Abstract (English) XIV Abstract (German) XVII I Theoretical Part 1 Introduction 1 1.1 Medical need in modern society 1 1.2 Natural products in drug development 1 1.3 Polycyclic polyprenylated acylphloroglucinols (PPAPs) 2 1.3.1 Classifications 2 1.3.2 Biosynthesis 4 1.3.3 Biological activities 6 1.3.4 PPAP-synthesis strategies 7 1.4 Flavonoids in drug discovery 10 1.5 Hybrid drugs in cancer treatment 11 1.5.1 Antibody-drug conjugates 11 1.5.2 Drug-drug conjugates 12 1.5.3 Prostate cancer and prostate-specific membrane antigen (PSMA) 13 2 Objectives 16 2.1 Extending the PPAP library and improving biological activities 16 2.2 Examination and improvement of anticancer efficacy with unexplored PPAP scaffolds 17 2.3 Improving target selectivity for PCa using PSMA-targeting PPAPs 17 3 Expanding the PPAP library 18 3.1 Evaluation of the existing library 18 3.2 Synthesis of the PPAP22 core (PPAP26, 31) 21 3.3 Acylation of the C3 position (head group) 24 3.3.1 Established acylation method using acyl cyanides 24 3.3.2 Cyanide free acylation by PESLALZ 25 3.4 Structure-activity relationship (SAR) study of PPAP78 (50) derivatives (PPAP generation 2) 26 3.4.1 Designing the SAR study 26 3.4.2 Synthesis of PPAPs for the SAR study 28 3.4.3 Evaluation of antibacterial activity 29 3.4.4 Conclusion 32 3.5 Evaluation of anticancer activity 33 3.5.1 Synthesis of the used PPAPs 33 3.5.2 Blood cancer (leukaemia) 36 3.5.3 Brain cancer (glioblastoma) 39 3.5.4 Prostate cancer (PCa) 42 3.6 Exploring synthesis methods towards new PPAP amides 46 3.6.1 PPAP amide synthesis via isocyanates 46 3.6.2 PPAP amides from carbamates 49 3.7 Summary of the library expansion 51 4 Novel flavonoid-like PPAP scaffold 54 4.1 Exploring approaches to flavonoid-like PPAPs 55 4.1.1 Flavanones in one step from PPAP26 (31) 55 4.1.2 Conversion of flavanones to flavones 58 4.1.3 Flavone and aurone synthesis with propiolic acids 59 4.1.4 First biological activity screening 62 4.1.5 Synthesis of substituted propiolic acids 63 4.1.6 Elaborating the substitution scope with dimedone test substrates 65 4.2 Investigating flavone-like PPAPs 66 4.2.1 Synthesis of flavone-like PPAPs 66 4.2.2 Biological activity investigations 68 4.3 Exploring the synthesis of isoflavanone-like PPAPs 71 4.4 Summary of flavonoid-like PPAPs 74 5 Studies towards PPAP hybrid drugs 76 5.1 PPAP drug-drug conjugates 76 5.1.1 PPAP–TMZ conjugates 77 5.1.2 PPAP–DOX conjugates 92 5.1.3 Summary of PPAP drug-drug conjugates 96 5.2 PPAP antibody-drug conjugates 97 5.2.1 Synthesis of a linker with a Val–Cit recognition sequence 99 5.2.2 Synthesis and coupling of simplified linkers 102 5.2.3 Testing of cysteine coupling potential 106 5.2.4 Summary of PPAP antibody-drug conjugates 107 5.3 Using PSMA to increase PCa specificity of PPAPs 107 5.3.1 In silico studies for PSMA-targeting PPAPs 108 5.3.2 Synthesis of PSMA-targeting PPAPs 115 5.3.3 Biological activity of PSMA-targeting PPAPs 217 – 220 118 5.3.4 Studies towards PSMA-targeting PPAPs with cleavable hydrazide linkers 121 5.3.5 Summary of PSMA-targeting PPAPs 127 6 Summary of results 129 II Experimental Part 7 General Remarks 137 7.1 Depiction of structures 137 7.2 Substance names 137 7.3 Solvents and common chemicals 137 7.4 Analytical methods 138 7.5 Biological tests 140 7.6 Docking studies with Schrödinger© 146 8 General synthesis procedures 147 9 Syntheses expanding the PPAP library 150 9.1 Synthesis of PPAP22-core (PPAP26, 31) 150 9.2 PPAP derivatisation at C3 161 9.2.1 Standard acyl derivatisation 161 9.2.2 Standard PPAP amide synthesis 197 9.2.3 Studies towards a new PPAP amide synthesis 201 9.3 PPAP salts 205 10 Synthesis of flavonoid-like PPAPs 209 10.1 Synthesis of substituted propiolic acids 209 10.1.1 Acetylene precursors 209 10.1.2 Substituted propiolic acids 211 10.2 Synthesis of flavonoid-like dimedone test-substrates 216 10.3 Synthesis of flavonoid-like PPAPs 223 10.3.1 Aurone-like PPAPs 223 10.3.2 Flavanoid- and chalcone-like PPAPs 224 10.3.3 Flavone-like PPAPs 227 10.3.4 Dihydrocoumarin-like PPAPs 240 11 Synthesis of PPAP hybrids 242 11.1 PPAP drug-drug conjugates 242 11.1.1 PPAP-Temozolomide conjugates 242 11.1.2 PPAP-Doxorubicin conjugates 267 11.2 PPAP-antibody conjugates 276 11.2.1 Linker synthesis 276 11.2.2 Conjugate synthesis and modification 284 11.3 PSMA-targeting PPAPs 293 11.3.1 Synthesis of linkable PSMA-targeting units (PSMA-TUs) 293 11.3.2 Synthesis of a linkable PPAP amide 300 11.3.3 Linking PSMA-TUs to PPAPs 301 11.3.4 Deprotection of tBu-esters 312 III Appendix 12 X-Ray crystal structures 317 13 Exact structures of docked ligands 341 14 References 345 Curriculum vitae 355 List of publications 356 / Im Zuge dieser Arbeit wurden Verbindungen der Naturstoffklasse der polyzyklischen polyprenylierten Acylphloroglucinole (PPAPs) untersucht. PPAPs besitzen eine polyzyklische Kernstruktur, die mehrere Carbonylgruppen aufweist und mit ungesättigten Seitenketten dekoriert ist. Die in der Arbeitsgruppe entwickelte und optimierte, modulare Syntheseplattform ermöglichte die Erstellung einer ersten PPAP-Bibliothek mit 23 Verbindungen. Damit wurde die großflächige Untersuchung der antibakteriellen Aktivitäten von PPAPs eingeleitet. Weitere Erforschungen zur Derivatisierung von PPAPs ebneten den Weg zu einem synthetischen Werkzeugkasten für die Erzeugung neuer Derivate. Eine nachfolgende Struktur-Aktivitäts-Beziehungsstudie (engl. structure-activity relationship, SAR), die durch eine neuartige, in der Gruppe entwickelte C-Acylierungsstrategie ermöglicht wurde, erweiterte die Bibliothek um 25 neue nicht natürlich vorkommende Typ B PPAPs. Hierbei konnten antibiotische Wirksamkeiten mit minimalen Hemmkonzentrationen (MICs) im nanomolaren Bereich gegen Methicillin-resistenten Staphylococcus aureus (MRSA) erreicht werden und neue Leitverbindungen mit großen therapeutischen Fenstern wurden identifiziert. Die krebshemmende Wirkung von PPAPs wurde an den verschiedenen Krebsarten Leukämie, Glioblastom (GB) und Prostatakrebs (PCa) untersucht. Hierbei erwies sich eine 3 Phenylpropanoyl-Kopfgruppe als entscheidend für die proapoptotische Wirksamkeit. Strukturoptimierungen zeigten, dass für jede Krebsart jeweils unterschiedliche Verbindungen potente Wirkungen mit proapoptotischen halbmaximal inhibierenden Konzentrationen (IC50) im niedrigen mikromolaren und antiproliferativen IC50-Werten im nanomolaren Bereich aufwiesen. Flavonoid-ähnliche PPAP-Gerüste wurden im Hinblick auf verstärkte biologische Aktivitäten durch die Kombination der krebshemmenden Aktivitäten von PPAPs mit Flavonoiden untersucht. Es konnte eine einstufige Synthesemethode etabliert werden, mit welcher elf Flavonoid-ähnliche PPAPs mit vielversprechendem Krebs-bekämpfungspotenzial, insbesondere gegen GB, hergestellt wurden. Sie zeigten eine starke proapoptotische Wirkung bei geringer Toxizität gegenüber gesunden Zellen. Um die kovalente Verknüpfung verschiedener Wirkstoffe miteinander zu erweitern, wurden zusätzliche Konjugationsstrategien für die Synthese von PPAP-haltigen Wirkstoff-Wirkstoff-Konjugaten (engl.: drug-drug conjugates, DDCs) eingesetzt. Dies führte zu mehreren PPAP–Temozolomid- (TMZ) und PPAP–Doxorubicin (DOX) Konjugaten. PPAP–TMZ zeigte eine signifikante krebshemmende Wirksamkeit hinsichtlich GB, während PPAP–DOX eine beträchtliche antileukämische Aktivität zeigte. Problematisch ist, dass beide DDCs auch erhebliche toxische Wirkungen auf gesunde periphere mononukleare Blutzellen (PBMCs) zeigten. Aus diesem Grund wurden Untersuchungen zur Verbesserung der Selektivität von PPAP-Behandlungen unter Zuhilfenahme von Wirkstoffhybriden für den gezielten Wirkstofftransport (engl.: targeted drug delivery) durchgeführt. Dazu wurden PPAPs mit Linkern für Antikörper-Wirkstoff-Konjugate (engl.: antibody-drug conjugate, ADCs) verknüpft. Um das Kopplungspotenzial des hergestellten Linkers mit Antikörpern zu testen, wurde dieser mit Cystein umgesetzt. Die erfolgreiche Reaktion zeigte das Potenzial von zukünftigen Kopplungen mit echten Antikörpern. Speziell für PCa wurden gegen das prostataspezifische Membranantigen (PSMA) gerichtete PPAPs erforscht. Dieses Protein wird ausschließlich von PCa-Zellen exprimiert und stellt daher eine ideale Zielstruktur dar. Die Verwendung chemischer Strukturen aus bereits eingesetzten Arzneimitteln führte zur Entwicklung verschiedener PSMA-gerichteter PPAPs. Während in-silico-Screenings auf eine außergewöhnlich hohe Affinität zur PSMA-Bindungsstelle hinwiesen, ergaben biologische Tests nur eine sehr geringe krebshemmende Wirkung. Aufgrund der zu starken Bindungsaffinität wurden die PPAPs in der Zelle vermutlich nicht freigesetzt, was die niedrige krebshemmende Wirkung erklären könnte. Deshalb wurden im Rahmen weiterer Forschung spaltbare Linkersysteme untersucht. Insgesamt hat diese Forschungsarbeit das Verständnis für verschiedene PPAP-Derivate erheblich verbessert und ihr Potenzial für antibakterielle und krebsbekämpfende Anwendungen aufgezeigt. Die skizzierten Synthesestrategien, SAR-Studien und Konjugationsansätze bilden eine Grundlage für zukünftige Weiterentwicklungen dieses Forschungsprojektes. Weitere Optimierungen und Kooperationen sind notwendig, um PPAPs einen Schritt näher an die Anwendung als medizinische Behandlungsoption zu bringen.:Table of Contents Acknowledgements III Table of Contents V List of Abbreviations IX Abstract (English) XIV Abstract (German) XVII I Theoretical Part 1 Introduction 1 1.1 Medical need in modern society 1 1.2 Natural products in drug development 1 1.3 Polycyclic polyprenylated acylphloroglucinols (PPAPs) 2 1.3.1 Classifications 2 1.3.2 Biosynthesis 4 1.3.3 Biological activities 6 1.3.4 PPAP-synthesis strategies 7 1.4 Flavonoids in drug discovery 10 1.5 Hybrid drugs in cancer treatment 11 1.5.1 Antibody-drug conjugates 11 1.5.2 Drug-drug conjugates 12 1.5.3 Prostate cancer and prostate-specific membrane antigen (PSMA) 13 2 Objectives 16 2.1 Extending the PPAP library and improving biological activities 16 2.2 Examination and improvement of anticancer efficacy with unexplored PPAP scaffolds 17 2.3 Improving target selectivity for PCa using PSMA-targeting PPAPs 17 3 Expanding the PPAP library 18 3.1 Evaluation of the existing library 18 3.2 Synthesis of the PPAP22 core (PPAP26, 31) 21 3.3 Acylation of the C3 position (head group) 24 3.3.1 Established acylation method using acyl cyanides 24 3.3.2 Cyanide free acylation by PESLALZ 25 3.4 Structure-activity relationship (SAR) study of PPAP78 (50) derivatives (PPAP generation 2) 26 3.4.1 Designing the SAR study 26 3.4.2 Synthesis of PPAPs for the SAR study 28 3.4.3 Evaluation of antibacterial activity 29 3.4.4 Conclusion 32 3.5 Evaluation of anticancer activity 33 3.5.1 Synthesis of the used PPAPs 33 3.5.2 Blood cancer (leukaemia) 36 3.5.3 Brain cancer (glioblastoma) 39 3.5.4 Prostate cancer (PCa) 42 3.6 Exploring synthesis methods towards new PPAP amides 46 3.6.1 PPAP amide synthesis via isocyanates 46 3.6.2 PPAP amides from carbamates 49 3.7 Summary of the library expansion 51 4 Novel flavonoid-like PPAP scaffold 54 4.1 Exploring approaches to flavonoid-like PPAPs 55 4.1.1 Flavanones in one step from PPAP26 (31) 55 4.1.2 Conversion of flavanones to flavones 58 4.1.3 Flavone and aurone synthesis with propiolic acids 59 4.1.4 First biological activity screening 62 4.1.5 Synthesis of substituted propiolic acids 63 4.1.6 Elaborating the substitution scope with dimedone test substrates 65 4.2 Investigating flavone-like PPAPs 66 4.2.1 Synthesis of flavone-like PPAPs 66 4.2.2 Biological activity investigations 68 4.3 Exploring the synthesis of isoflavanone-like PPAPs 71 4.4 Summary of flavonoid-like PPAPs 74 5 Studies towards PPAP hybrid drugs 76 5.1 PPAP drug-drug conjugates 76 5.1.1 PPAP–TMZ conjugates 77 5.1.2 PPAP–DOX conjugates 92 5.1.3 Summary of PPAP drug-drug conjugates 96 5.2 PPAP antibody-drug conjugates 97 5.2.1 Synthesis of a linker with a Val–Cit recognition sequence 99 5.2.2 Synthesis and coupling of simplified linkers 102 5.2.3 Testing of cysteine coupling potential 106 5.2.4 Summary of PPAP antibody-drug conjugates 107 5.3 Using PSMA to increase PCa specificity of PPAPs 107 5.3.1 In silico studies for PSMA-targeting PPAPs 108 5.3.2 Synthesis of PSMA-targeting PPAPs 115 5.3.3 Biological activity of PSMA-targeting PPAPs 217 – 220 118 5.3.4 Studies towards PSMA-targeting PPAPs with cleavable hydrazide linkers 121 5.3.5 Summary of PSMA-targeting PPAPs 127 6 Summary of results 129 II Experimental Part 7 General Remarks 137 7.1 Depiction of structures 137 7.2 Substance names 137 7.3 Solvents and common chemicals 137 7.4 Analytical methods 138 7.5 Biological tests 140 7.6 Docking studies with Schrödinger© 146 8 General synthesis procedures 147 9 Syntheses expanding the PPAP library 150 9.1 Synthesis of PPAP22-core (PPAP26, 31) 150 9.2 PPAP derivatisation at C3 161 9.2.1 Standard acyl derivatisation 161 9.2.2 Standard PPAP amide synthesis 197 9.2.3 Studies towards a new PPAP amide synthesis 201 9.3 PPAP salts 205 10 Synthesis of flavonoid-like PPAPs 209 10.1 Synthesis of substituted propiolic acids 209 10.1.1 Acetylene precursors 209 10.1.2 Substituted propiolic acids 211 10.2 Synthesis of flavonoid-like dimedone test-substrates 216 10.3 Synthesis of flavonoid-like PPAPs 223 10.3.1 Aurone-like PPAPs 223 10.3.2 Flavanoid- and chalcone-like PPAPs 224 10.3.3 Flavone-like PPAPs 227 10.3.4 Dihydrocoumarin-like PPAPs 240 11 Synthesis of PPAP hybrids 242 11.1 PPAP drug-drug conjugates 242 11.1.1 PPAP-Temozolomide conjugates 242 11.1.2 PPAP-Doxorubicin conjugates 267 11.2 PPAP-antibody conjugates 276 11.2.1 Linker synthesis 276 11.2.2 Conjugate synthesis and modification 284 11.3 PSMA-targeting PPAPs 293 11.3.1 Synthesis of linkable PSMA-targeting units (PSMA-TUs) 293 11.3.2 Synthesis of a linkable PPAP amide 300 11.3.3 Linking PSMA-TUs to PPAPs 301 11.3.4 Deprotection of tBu-esters 312 III Appendix 12 X-Ray crystal structures 317 13 Exact structures of docked ligands 341 14 References 345 Curriculum vitae 355 List of publications 356

Polycyclic polyprenylated acylphloroglucinols (PPAPs) are structurally complex natural products with promising biological activities. These compounds have interesting anticancer and anti-HIV properties as well as other biological activities making them highly attractive synthetic targets. We report a stereodivergent, asymmetric total synthesis of (−)-clusianone in six steps from commercial materials. We have implemented a challenging cationic cyclization forging a bond between two sterically encumbered quaternary carbon atoms. Mechanistic studies point to the unique ability of formic acid to mediate the cyclization forming the clusianone framework. We also present a biosynthesis-inspired, diversity-oriented synthesis approach for rapid construction of PPAP analogs via palladium-catalyzed dearomative conjunctive allylic alkylation (DCAA). These efficient palladium-catalyzed protocols construct the [3.3.1]-bicyclic PPAP core in a single step from their stable aromatic precursors. The first syntheses of 13,14-didehydroxyisogarcinol and garcimultiflorone A stereoisomers are reported in six steps from a commercially available phloroglucinol. Lewis acid-controlled, diastereoselective cationic oxycyclizations enabled asymmetric syntheses of (−)-6-epi-13,14-didehydroxyisogarcinol and (+)-30-epi-13,14-didehydroxyisogarcinol. A similar strategy enabled production of the meso-derived isomers (±)-6,30-epi-13,14-didehydroxyisogarcinol and (±)-6,30-epi-garcmultiflorone A. A convenient strategy for gram scale synthesis of these stereoisomers was developed utilizing diastereomer separation at a later stage in the synthesis that minimized the number of necessary synthetic operations to access all possible stereoisomers. Finally, we report cationic rearrangements of dearomatized acylphloroglucinols leading to the formation of unprecedented PPAP scaffolds. A novel type A [3.3.1]-bicyclic PPAP was produced as a major product and the structure confirmed by X-ray crystallographic analysis. A novel [3.3.1]-bicyclolactone was also produced utilizing an alternative substrate. Efforts will be described to determine the scope of these rearrangements and type A-selective cyclizations. / 2018-08-09T00:00:00Z

<p>This report is a part of the education at the School of Engineering, Jönköping. The content of the report is based on work conducted at Huskvarna Prototyper AB (HPAB). The task of the report is to find out what reasonable quality demands would be for a short run manufacturer and what short run manufactures should do to satisfy the demands.</p><p>The car manufacturing industry is currently setting high quality demands through QS 9000, Production Part Approval Process (PPAP), on their suppliers. They have to show that they have reliable processes and no quality failures on supplied parts. Statistical process control (SPC) methods are used to comply with the demands set. SPC helps the suppliers to secure that their products are shifting within a small interval. Suppliers can with the help of control charts with a upper control limit and a lower control limit see if their process needs adjustment.</p><p>The car manufacturing industry demands that their suppliers have stable processes. In order to show that the processes are stable a Cpk value is used. It has to be greater than 1.33 to assure a stable process.</p><p>HPAB has got problems with a component called Scuff Plate (a part of Volvo XC 90) which has got its standards set by the car manufacturing industry. The authors of this paper have measured the Cpk value at four different occasions with the help of the measurement program, Rektron. The value on observation 1, was 0.15 but has increased to 1.34, on observation 4, thanks to adjustments made.</p><p>The autors have produced an instruction manual for dealing with future short and long run production.</p> / <p>Denna rapport är en del i en högskoleingenjörsutbildning vid Tekniska Högskolan i Jönköping. Arbetet är utfört på Huskvarna Prototyper AB (HPAB). Uppgiften var att ta reda på vilka kvalitetskrav som är rimliga att ställa på småserietillverkare och vad småserietillverkarna kan göra för att uppfylla kraven.</p><p>Bilindustrin ställer i dag höga krav på sina underleverantörer. De ska kunna visa att de har dugliga processer och på de detaljer de levererar ska det inte finnas några kvalitetsbrister. För att säkerställa hög kvalité jobbar många företag med statistiska metoder där man med hög säkerhet kan visa att ens måttvariation skiftar inom ett litet intervall.</p><p>Statistisk processstyrning (SPS) är ett hjälpmedel för att säkerhetsställa god kvalité. Genom regelbundna mätningar får man fram värden som man för in i ett styrdiagram. Med hjälp av detta styrdiagram, som består av en övre och en undre styrgräns, kan man avläsa när processen måste justeras.</p><p>Bilindustrin kräver att deras underleverantörer har stabila processer. Detta kan man visa genom att ta fram ett korrigeradat maskinduglighetsvärde, Cpk. QS 9000 med dess Production Part Approval Process (PPAP), är den standard som har tagits fram av bilindustrin kräver att processerna ska ha ett Cpk som är större än 1,33 för att processen ska vara godkänd.</p><p>HPAB har problem med detaljen Scuff Plate som är en detalj till Volvo XC90. Vid fyra olika tillfällen har maskinduglighetstest utförst på HPAB:s quintusspress. I rapporten kan man följa hur Cpk-värdet vid första observationen på 0,15 har ökat till 1,34 vid det fjärde testet. Att Cpk-värdet har ökat beror på att man ändrat inställningar mellan de olika observationstillfällena.</p><p>Författarna har tagit fram en mall för hur HPAB, som mestadels tillverkar små volymer, ska hantera de krav som bilindustrin ställer vid både långa och korta serier.</p>

This report is a part of the education at the School of Engineering, Jönköping. The content of the report is based on work conducted at Huskvarna Prototyper AB (HPAB). The task of the report is to find out what reasonable quality demands would be for a short run manufacturer and what short run manufactures should do to satisfy the demands. The car manufacturing industry is currently setting high quality demands through QS 9000, Production Part Approval Process (PPAP), on their suppliers. They have to show that they have reliable processes and no quality failures on supplied parts. Statistical process control (SPC) methods are used to comply with the demands set. SPC helps the suppliers to secure that their products are shifting within a small interval. Suppliers can with the help of control charts with a upper control limit and a lower control limit see if their process needs adjustment. The car manufacturing industry demands that their suppliers have stable processes. In order to show that the processes are stable a Cpk value is used. It has to be greater than 1.33 to assure a stable process. HPAB has got problems with a component called Scuff Plate (a part of Volvo XC 90) which has got its standards set by the car manufacturing industry. The authors of this paper have measured the Cpk value at four different occasions with the help of the measurement program, Rektron. The value on observation 1, was 0.15 but has increased to 1.34, on observation 4, thanks to adjustments made. The autors have produced an instruction manual for dealing with future short and long run production. / Denna rapport är en del i en högskoleingenjörsutbildning vid Tekniska Högskolan i Jönköping. Arbetet är utfört på Huskvarna Prototyper AB (HPAB). Uppgiften var att ta reda på vilka kvalitetskrav som är rimliga att ställa på småserietillverkare och vad småserietillverkarna kan göra för att uppfylla kraven. Bilindustrin ställer i dag höga krav på sina underleverantörer. De ska kunna visa att de har dugliga processer och på de detaljer de levererar ska det inte finnas några kvalitetsbrister. För att säkerställa hög kvalité jobbar många företag med statistiska metoder där man med hög säkerhet kan visa att ens måttvariation skiftar inom ett litet intervall. Statistisk processstyrning (SPS) är ett hjälpmedel för att säkerhetsställa god kvalité. Genom regelbundna mätningar får man fram värden som man för in i ett styrdiagram. Med hjälp av detta styrdiagram, som består av en övre och en undre styrgräns, kan man avläsa när processen måste justeras. Bilindustrin kräver att deras underleverantörer har stabila processer. Detta kan man visa genom att ta fram ett korrigeradat maskinduglighetsvärde, Cpk. QS 9000 med dess Production Part Approval Process (PPAP), är den standard som har tagits fram av bilindustrin kräver att processerna ska ha ett Cpk som är större än 1,33 för att processen ska vara godkänd. HPAB har problem med detaljen Scuff Plate som är en detalj till Volvo XC90. Vid fyra olika tillfällen har maskinduglighetstest utförst på HPAB:s quintusspress. I rapporten kan man följa hur Cpk-värdet vid första observationen på 0,15 har ökat till 1,34 vid det fjärde testet. Att Cpk-värdet har ökat beror på att man ändrat inställningar mellan de olika observationstillfällena. Författarna har tagit fram en mall för hur HPAB, som mestadels tillverkar små volymer, ska hantera de krav som bilindustrin ställer vid både långa och korta serier.

Gold catalysis has become an important tool to achieve highly chemoselective p-acid activation. Exceptional reactivity and selectivity are often encountered under mild reaction conditions. These properties have made gold(I) complexes suitable catalysts for tremendous applications in the total synthesis of natural products. The first chapter will highlight a number of total syntheses using gold catalysis as a key step. The second chapter will cover our application of the gold(I)-catalyzed 6-endo-dig carbocyclization for the synthesis of bridgehead-substituted scaffolds and its use toward the synthesis of PPAP natural products. This research has opened our eyes to the utility of biphenylphosphine ligands, particularly JohnPhos, in gold(I)-catalysis. The reactivity and selectivity exhibited by gold(I) complexes is modulated by the nature of the ancillary ligand. Recent research rationalizes the impact of these ligands on the divergent reactivity observed between cationic and carbenoid intermediates. Our desire to favor the 6-endo-dig pathway has led us toward the discovery of another example of the diagonal reactivity that NHC carbene and biphenylphosphine ligands can bring to gold(I)-catalysis. Chapter three will explain the development of a selective gold-catalyzed synthesis of fused carbocycles . Our selective divergent synthesis of fused carbocycles, combined with the Diels–Alder reaction, has brought new synthetic opportunities. Chapter four will describe our approach toward the synthesis of various polycyclic diterpene-related frameworks. Starting with a unique linear precursor, we have developed a new “one-pot” process for the synthesis of three different polycyclic compounds related to the terpenoid family. The facile modulation of the linear precursor and the use of different dienophiles during the Diels–Alder reaction could enable the synthesis of diverse polycyclic analogues based on three principal frameworks. The gold(I)-catalyzed synthesis of fused carbocycles reached some limitations during our study. Regioselective control was found to be substantially more challenging, with terminal alkynes or alkynes bearing a sterically and electronically neutral methyl substituent. In chapter five, we will discuss how the complementarity of silver(I) catalysis to gold(I) catalysis enabled the selective divergent synthesis of three different fused carbocycles from a unique precursor. Moreover, copper(I) catalysis has given access to the 6-endo-dig pathway on terminal alkynes without the formation of a vinylidene intermediate.