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  • 101. Ferrins, Lori
    et al.
    Gazdik, Michelle
    Rahmani, Raphael
    Varghese, Swapna
    Sykes, Melissa L.
    Jones, Amy J.
    Avery, Vicky M.
    White, Karen L.
    Ryan, Eileen
    Charman, Susan A.
    Kaiser, Marcel
    Bergstroem, Christel A. S.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Baell, Jonathan B.
    Pyridyl Benzamides as a Novel Class of Potent Inhibitors for the Kinetoplastid Trypanosoma brucei2014In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 57, no 15, p. 6393-6402Article in journal (Refereed)
    Abstract [en]

    A whole-organism screen of approximately 87000 compounds against Trypanosoma brucei brucei identified a number of promising compounds for medicinal chemistry optimization. One of these classes of compounds we termed the pyridyl benzamides. While the initial hit had an IC50 of 12 mu M, it was small enough to be attractive for further optimization, and we utilized three parallel approaches to develop the structure-activity relationships. We determined that the physicochemical properties for this class are generally favorable with particular positions identified that appear to block metabolism when substituted and others that modulate solubility. Our most active compound is 79, which has an IC50 of 0.045 mu M against the human pathogenic strain Trypanosoma brucei rhodesiense and is more than 4000 times less active against the mammalian L6 cell line.

  • 102.
    Fransson, Rebecca
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Discovery of Small Peptides and Peptidomimetics Targeting the Substance P 1-7 Binding Site: Focus on Design, Synthesis, Structure-Activity Relationships and Drug-Like Properties2011Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Biologically active peptides are important for many physiological functions in the human body and therefore serve as interesting starting points in drug discovery processes. In this work the neuropeptide substance P 1–7 (SP1–7, H-Arg-Pro-Lys-Pro-Gln-Gln-Phe-OH), which has been demonstrated to reduce neuropathic pain and attenuate opioid withdrawal symptoms in animal models, has been addressed in a medicinal chemistry program with the overall aim of transforming this bioactive peptide into more drug-like compounds. Specific binding sites for this neuropeptide have been detected in the brain and the spinal cord. Interestingly, the smaller neuropeptide endomorphin-2 (EM-2, H-Tyr-Pro-Phe-Phe-NH2) also interacts with these binding sites, although 10-fold less efficient.

    In this work the structure–activity relationship of SP1–7 and EM-2, regarding their affinity to the SP1–7 binding site was elucidated using alanine scans, truncation, and terminal modifications. The C-terminal part of both peptides, and especially the C-terminal phenylalanine, was crucial for binding affinity. Moreover, the C-terminal functional group should preferably be a primary amide. The truncation studies finally resulted in the remarkable discovery of H-Phe-Phe-NH2 as an equally good binder as the heptapeptide SP1–7. This dipeptide amide served as a lead compound for further studies. In order to improve the drug-like properties and to find a plausible bioactive conformation, a set of rigidified and methylated dipeptides of different stereochemistry, and analogs with reduced peptide character, were synthesized and evaluated regarding binding, metabolic stability and absorption. Small SP1–7 analogs with retained affinity and substantially improved permeability and metabolic stability were identified.

    Beside peptide chemistry the synthetic work included the development of a fast and convenient microwave-assisted protocol for direct arylation of imidazoles. Furthermore, microwave-assisted aminocarbonylation using Mo(CO)6 as a solid carbon monoxide source was investigated in the synthesis of MAP amides and for coupling of imidazoles with amino acids.

    In a future perspective the present findings, together with the fact that some of the SP1–7 analogs discovered herein have been shown to reproduce the biological effects of SP1-7 in animal studies related to neuropathic pain and opioid dependence, can ultimately have an impact on drug discovery in these two areas.

    List of papers
    1. Small peptides mimicking substance P (1-7) and encompassing a C-terminal amide functionality
    Open this publication in new window or tab >>Small peptides mimicking substance P (1-7) and encompassing a C-terminal amide functionality
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    2008 (English)In: Neuropeptides, ISSN 0143-4179, E-ISSN 1532-2785, Vol. 42, no 1, p. 31-37Article in journal (Refereed) Published
    Abstract [en]

    Some of the biological effects demonstrated after administration of substance P (SP) in vivo can indirectly be attributed to the fragmentation of the undecapeptide to its N-terminal bioactive fragment SP1–7. This heptapeptide (H-Arg-Pro-Lys-Pro-Gln-Gln-Phe-OH) is a major bioactive metabolite from SP that frequently exerts similar biological effects as the parent peptide but also, in several cases, completely opposite actions. Specific binding sites for the heptapeptide SP1–7 that are separate from the SP preferred NK receptors have been identified. In this study we demonstrate that (a) the C-terminal part of the SP metabolite SP1–7 is most important for binding as deduced from an Ala scan and that a replacement of Phe7 for Ala is deleterious, (b) truncation of the N-terminal amino acid residues of SP1–7 delivers peptides with retained binding activity, although with somewhat lower binding affinities than SP1–7 and (c) a C-terminal amide group as a replacement for the terminal carboxy group of SP1–7 and for all of the truncated ligands synthesized affords approximately 5–10-fold improvements of the binding affinities.

    Keywords
    Substance P (1–7), SP (1–7), SP1–7, Substance P, Structure–activity relationships (SAR), Ala scan, Peptidomimetics
    National Category
    Pharmaceutical Sciences
    Identifiers
    urn:nbn:se:uu:diva-97823 (URN)10.1016/j.npep.2007.11.002 (DOI)000254231100003 ()18093649 (PubMedID)
    Available from: 2008-11-21 Created: 2008-11-21 Last updated: 2018-01-13Bibliographically approved
    2. Discovery of Dipeptides with High Affinity to the Specific Binding Site for Substance P1-7
    Open this publication in new window or tab >>Discovery of Dipeptides with High Affinity to the Specific Binding Site for Substance P1-7
    Show others...
    2010 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 53, no 6, p. 2383-2389Article in journal (Refereed) Published
    Abstract [en]

    Substance P 1-7 (SP1-7, H-Arg-Pro-Lys-Pro-Gln-Gln-Phe-OH) is the major bioactive metabolite of substance P. The interest in this heptapeptide originates from the observation that it modulates, and in certain cases opposes the effects of the parent peptide. e.g., the nociceptive effect. The p-opioid receptor agonist endomorphin-2 (EM-2, H-Tyr-Pro-Phe-Phe-NH2) has been found to also interact with the specific binding site of SP1-7 with only a 10-fold lower affinity compared to the native peptide. Considering the smaller size of EM-2 compared to the target heptapeptide, it was selected as a lead compound in the development of low-molecular-weight ligands to the SP1-7 binding site. An alanine scan and truncation study led to the unexpected discovery of the dipeptide H-Phe-Phe-NH2 (K-i = 1.5 nM), having equal affinity as the endogenous heptapeptide SP1-7. Moreover, the studies show that the C-terminal phenylalanine amide is crucial for the affinity of the dipeptide.

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-136801 (URN)10.1021/jm901352b (DOI)000275711000006 ()20178322 (PubMedID)
    Available from: 2010-12-14 Created: 2010-12-14 Last updated: 2017-12-11Bibliographically approved
    3. Constrained H-Phe-Phe-NH2 Analogues With High Affinity to the Substance P 1-7 Binding Site and With Improved Metabolic Stability and Cell Permeability
    Open this publication in new window or tab >>Constrained H-Phe-Phe-NH2 Analogues With High Affinity to the Substance P 1-7 Binding Site and With Improved Metabolic Stability and Cell Permeability
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    2013 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 56, no 12, p. 4953-4965Article in journal (Refereed) Published
    Abstract [en]

    We recently reported the discovery of H-Phe-Phe-NH2 as a small and high affinity ligand for the substance P 1-7 (SP1-7, H-Arg-Pro-Lys-Pro-Gln-Gln-Phe-OH) specific binding site and its intriguing ability to reduce neuropathic pain. With the overall aim to develop stable and orally bioavailable SP1-7 mimetics, the dipeptide was chosen as a lead compound. Herein the structure-activity relationship (SAR) of a set of modified H-Phe-Phe-NH2 analogues is presented together with their potential active uptake by PEPT1 transporter, intestinal permeability, and metabolic stability. Local constraints via peptide backbone methylation or preparation of cyclized analogues based on pyrrolidine were evaluated and were shown to significantly improve the in vitro pharmacokinetic properties. The SAR was rationalized by deriving a plausible binding pose for the high affinity ligands. Rigidification using a 3-phenylpyrrolidine moiety in the C-terminal of H-Phe-Phe-NH2 resulted in high affinity and improved intrinsic clearance and intestinal epithelial permeability.

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-149482 (URN)10.1021/jm400209h (DOI)000321237100011 ()23735006 (PubMedID)
    Available from: 2011-03-24 Created: 2011-03-20 Last updated: 2017-12-11Bibliographically approved
    4. Discovery and Pharmacokinetic Profiling of Phenylalanine Based Carbamates as Novel Substance P 1-7 Analogues
    Open this publication in new window or tab >>Discovery and Pharmacokinetic Profiling of Phenylalanine Based Carbamates as Novel Substance P 1-7 Analogues
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    (English)Manuscript (preprint) (Other academic)
    Identifiers
    urn:nbn:se:uu:diva-149995 (URN)
    Available from: 2011-03-24 Created: 2011-03-24 Last updated: 2011-05-03
    5. Design and Synthesis of N-Terminal Imidazole Based H-Phe-Phe-NH2 Mimetics
    Open this publication in new window or tab >>Design and Synthesis of N-Terminal Imidazole Based H-Phe-Phe-NH2 Mimetics
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    (English)Manuscript (preprint) (Other academic)
    Identifiers
    urn:nbn:se:uu:diva-149996 (URN)
    Available from: 2011-03-24 Created: 2011-03-24 Last updated: 2011-05-03
    6. Microwave-Assisted Synthesis of Weinreb and MAP Aryl Amides via Pd-Catalyzed Heck Aminocarbonylation Using Mo(CO)(6) or W(CO)(6)
    Open this publication in new window or tab >>Microwave-Assisted Synthesis of Weinreb and MAP Aryl Amides via Pd-Catalyzed Heck Aminocarbonylation Using Mo(CO)(6) or W(CO)(6)
    2011 (English)In: Journal of Organic Chemistry, ISSN 0022-3263, E-ISSN 1520-6904, Vol. 76, no 3, p. 978-981Article in journal (Refereed) Published
    Abstract [en]

    A simple and expedient process for the Heck aminocarbonylative synthesis of Weinreb and MAP amide acylating agents, from aryl halides, is reported. This methodology utilizes solid sources of CO making it readily accessible to chemists working in small-scale laboratory applications.

    National Category
    Pharmaceutical Sciences
    Identifiers
    urn:nbn:se:uu:diva-148925 (URN)10.1021/jo102151u (DOI)000286577400027 ()
    Available from: 2011-03-15 Created: 2011-03-14 Last updated: 2018-01-12Bibliographically approved
  • 103.
    Friggeri, Laura
    et al.
    Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Ballante, Flavio
    Rome Center for Molecular Design Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Ragno, Rino
    Rome Center for Molecular Design Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Musmuca, Ira
    Rome Center for Molecular Design Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    De Vita, Daniela
    Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Manetti, Fabrizio
    Dipartimento di Biotecnologie, Chimica e Farmacia, Universita ̀ degli Studi di Siena, Via Aldo Moro 2, I-53100 Siena, Italy.
    Biava, Mariangela
    Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Scipione, Luigi
    Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Di Santo, Roberto
    Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Costi, Roberta
    Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Feroci, Marta
    Dipartimento di Scienze di Base e Applicate per l ’ Ingegneria, Sapienza Universita ̀ di Roma, Via Castro Laurenziano 7, I-00161 Rome, Italy.
    Tortorella, Silvano
    Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Universita ̀ di Roma, P. le A. Moro 5, 00185 Rome, Italy.
    Pharmacophore assessment through 3-D QSAR: evaluation of the predictive ability on new derivatives by the application on a series of antitubercular agents.2013In: Journal of Chemical Information and Modeling, ISSN 1549-9596, E-ISSN 1549-960X, Vol. 53, no 6, p. 1463-74Article in journal (Refereed)
    Abstract [en]

    Pharmacophoric mapping is a useful procedure to frame, especially when crystallographic receptor structures are unavailable as in ligand-based studies, the hypothetical site of interaction. In this study, 71 pyrrole derivatives active against M. tuberculosis were used to derive through a recent new 3-D QSAR protocol, 3-D QSAutogrid/R, several predictive 3-D QSAR models on compounds aligned by a previously reported pharmacophoric application. A final multiprobe (MP) 3-D QSAR model was then obtained configuring itself as a tool to derive pharmacophoric quantitative models. To stress the applicability of the described models, an external test set of unrelated and newly synthesized series of R-4-amino-3-isoxazolidinone derivatives found to be active at micromolar level against M. tuberculosis was used, and the predicted bioactivities were in good agreement with the experimental values. The 3-D QSAutogrid/R procedure proved to be able to correlate by a single multi-informative scenario the different activity molecular profiles thus confirming its usefulness in the rational drug design approach.

  • 104.
    Gallo-Payet, Nicole
    et al.
    Service of Endocrinology, Department of Medicine, University of Sherbrooke, Canada.
    Guimond, Marie-Odile
    Service of Endocrinology, Department of Medicine, University of Sherbrooke, Canada.
    Bilodeau, Lyne
    Service of Endocrinology, Department of Medicine, University of Sherbrooke, Canada.
    Wallinder, Charlotta
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Alterman, Mathias
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Hallberg, Anders
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Angiotensin II, a neuropeptide at frontier between endocrinology and neuroscience:  is there a link between the angiotensin II type 2 receptor andAlzheimer’s disease?2011In: Frontiers in Endocrinology, ISSN 1664-2392, Vol. 2, p. Article 17-Article in journal (Refereed)
    Abstract [en]

    Amyloid-β peptide deposition, abnormal hyperphosphorylation of tau, as well as inflammation and vascular damage, are associated with the development of Alzheimer’s disease (AD). Angiotensin II (Ang II) is a peripheral hormone, as well as a neuropeptide, which binds two major receptors, namely the Ang II type 1 receptor (AT1R) and the type 2 receptor (AT2R). Activation of the AT2R counteracts most of the AT1R-mediated actions, promoting vasodilation, decreasing the expression of pro-inflammatory cytokines, both in the brain and in the cardiovascular system. There is evidence that treatment with AT1R blockers (ARBs) attenuates learning and memory deficits. Studies suggest that the therapeutic effects of ARBs may reflect this unopposed activation of the AT2R in addition to the inhibition of the AT1R. Within the context of AD, modulation of AT2R signaling could improve cognitive performance not only through its action on blood flow/brain microcirculation but also through more specific effects on neurons. This review summarizes the current state of knowledge and potential therapeutic relevance of central actions of this enigmatic receptor. In particular, we highlight the possibility that selective AT2R activation by non-peptide and highly selective agonists, acting on neuronal plasticity, could represent new pharmacological tools that may help improve impaired cognitive performance in AD and other neurological cognitive disorders.

  • 105.
    Georgsson, Jennie
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Design and Synthesis of Novel AT2 Receptor Ligands: From Peptides to Drug-Like Molecules2006Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Many peptide receptors are of pharmaceutical interest and there is thus a need for new ligands for such receptors. Unfortunately, peptides are not suitable as orally administrated drugs since they are associated with poor absorption, rapid metabolism and low sub-receptor selectivity. One approach that should allow identification of more drug-like ligands is to use the structural information of the endogenous ligand to develop peptidomimetic compounds.

    The main objective of the work described in this thesis was to convert angiotensin II (Ang II, Asp-Arg-Val-Tyr-Ile-His-Pro-Phe) to small drug-like compounds with retained bioactivity at the AT2 receptor. The study was performed step-wise via incorporation of well-defined secondary structure mimetics and repeated truncation of the peptide. Five scaffolds, comprising a benzene ring as a central element, suitable as a γ-turn or dipeptide mimetics were designed and synthesized. In order to decorate the scaffolds, a method of microwave-assisted alkoxycarbonylation was developed. After incorporation of the scaffolds into Ang II-related peptides or peptide fragments, the affinities for both the AT1 and the AT2 receptor were determined.

    In the first series of ligands, two tyrosine-related scaffolds were introduced as γ-turn mimetics in Ang II. All five pseudopeptides exhibited good affinities for the AT2 receptor. One compound was chosen for functional studies and was shown to act as an AT2 receptor agonist. After truncation of Ang II it was shown that C-terminal pentapeptide analogs were AT2 receptor selective agonists. A series of pseudopeptides comprising tyrosine-related scaffolds, derived from the pentapeptides, displayed high AT2 receptor affinities. Two compounds had agonistic effect at the AT2 receptor.

    This study revealed that the N-terminal part was of less importance while a C-terminal Ile residue was a key element for enhanced AT2 receptor affinity. In the final set of compounds, the peptide was truncated to tripeptide C-terminal fragments. After replacing His-Pro by a histidine-related scaffold small drug-like peptidomimetic compounds with nanomolar affinity for the AT2 receptor were identified.

    List of papers
    1. Rapid palladium-catalyzed synthesis of esters from aryl halides utilizing Mo(CO)6 as a solid carbon monoxide source
    Open this publication in new window or tab >>Rapid palladium-catalyzed synthesis of esters from aryl halides utilizing Mo(CO)6 as a solid carbon monoxide source
    2003 (English)In: Journal of Combinatorial Chemistry, Vol. 5, no 4, p. 350-352Article in journal (Refereed) Published
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-93926 (URN)
    Available from: 2006-01-13 Created: 2006-01-13 Last updated: 2013-07-04Bibliographically approved
    2. Angiotensin II Pseudopeptides Containing 1,3,5-Trisubstituted Benzene Scaffolds with High AT2 Receptor Affinity
    Open this publication in new window or tab >>Angiotensin II Pseudopeptides Containing 1,3,5-Trisubstituted Benzene Scaffolds with High AT2 Receptor Affinity
    Show others...
    2005 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 48, no 21, p. 6620-6631Article in journal (Refereed) Published
    Abstract [en]

    Two 1,3,5-trisubstituted aromatic scaffolds intended to serve as γ-turn mimetics have been synthesized and incorporated in five pseudopeptide analogues of angiotensin II (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe), replacing Val-Tyr-Ile, Val-Tyr, or Tyr-Ile. All the tested compounds exhibited nanomolar affinity for the AT2 receptor with the best compound (3) having a Ki of 1.85 nM. Four pseudopeptides were AT2 selective, while one (5) also exhibited good affinity for the AT1 receptor (Ki = 30.3 nM). This pseudopeptide exerted full agonistic activity in an AT2 receptor induced neurite outgrowth assay but displayed no agonistic effect in an AT1 receptor functional assay. Molecular modeling, using the program DISCOtech, showed that the high-affinity ligands could interact similarly with the AT2 receptor as other ligands with high affinity for this receptor. A tentative agonist model is proposed for AT2 receptor activation by angiotensin II analogues. We conclude that the 1,3,5-trisubstituted benzene rings can be conveniently prepared and are suitable as γ-turn mimics.

    National Category
    Natural Sciences
    Identifiers
    urn:nbn:se:uu:diva-95768 (URN)10.1021/jm050280z (DOI)
    Available from: 2007-04-17 Created: 2007-04-17 Last updated: 2017-12-14Bibliographically approved
    3. Short pseudopeptides containing turn scaffolds with high AT(2) receptor affinity
    Open this publication in new window or tab >>Short pseudopeptides containing turn scaffolds with high AT(2) receptor affinity
    Show others...
    2006 (English)In: Bioorganic & Medicinal Chemistry, ISSN 0968-0896, E-ISSN 1464-3391, Vol. 14, no 17, p. 5963-5972Article in journal (Refereed) Published
    Abstract [en]

    Two pentapeptides, Ac-Tyr-Ile-His-Pro-Phe/Ile, were synthesized and shown to have angiotensin II AT(2) receptor affinity and agonistic activity. Based on these peptides, a new series of 13 pseudopeptides was synthesized via introduction of five different turn scaffolds replacing the Tyr-Ile amino acid residues. Pharmacological evaluation disclosed subnanomolar affinities for some of these compounds at the AT(2) receptor. Substitution of Phe by Ile in this series of ligands enhanced the AT(2) receptor affinity of all compounds. These results suggest that the C-terminal amino acid residues can be elaborated on to enhance the AT(2) receptor affinity in truncated Ang II analogues.

    Keywords
    angiotensin II, AT1 receptor, AT2 receptor, AT2 receptor agonist, pseudopeptides, turn scaffolds
    National Category
    Pharmaceutical Sciences
    Identifiers
    urn:nbn:se:uu:diva-93928 (URN)10.1016/j.bmc.2006.05.019 (DOI)000239947500016 ()16753301 (PubMedID)
    Available from: 2006-01-13 Created: 2006-01-13 Last updated: 2018-01-13Bibliographically approved
    4. Synthesis of a new class of druglike angiotensin II C-terminal mimics with affinity for the AT2 receptor
    Open this publication in new window or tab >>Synthesis of a new class of druglike angiotensin II C-terminal mimics with affinity for the AT2 receptor
    Show others...
    2007 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 50, no 7, p. 1711-1715Article in journal (Refereed) Published
    Abstract [en]

    Four tripeptides corresponding to the C-terminal region of angiotensin II were synthesized. One of these peptides (Ac-His-Pro-Ile) showed moderate binding affinity for the AT2 receptor. Two aromatic histidine-related scaffolds were synthesized and introduced in the tripeptides to give eight new peptidomimetic structures. Three of the new peptide-derived druglike molecules exhibited selective, nanomolar affinity for the AT2 receptor. These ligands may become lead compounds in the future development of novel classes of selective AT2 receptor agonists.

    National Category
    Pharmaceutical Sciences
    Identifiers
    urn:nbn:se:uu:diva-95771 (URN)10.1021/jm0613469 (DOI)000245259000033 ()17358051 (PubMedID)
    Available from: 2007-04-17 Created: 2007-04-17 Last updated: 2018-01-13Bibliographically approved
  • 106.
    Ghasemzadeh, Nasim
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
    Application of Artificial Gel Antibodies for the Detection and Quantification of Proteins in Biological Fluids2010Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The molecular-imprinting method has previously been used for the synthesis of artificial gel antibodies, highly selective for various proteins. In present study, we have synthesized artificial gel antibodies against haemoglobin, albumin and different forms of growth hormone with the aim to develop a simple and rapid procedure to measure the concentration of these protein biomarkers in samples of clinical interest.  A spectrophotometric method was developed to design a standard curve in the form of a straight line, whereby the true absorption (not the recorded “apparent” absorption) was plotted against a known protein concentration. The procedure, applied to quantitative analysis of albumin in human plasma and cerebrospinal fluid (CSF) from patients with ALS, indicated that  the concentration of this protein was significantly enhanced in CSF from patients with amyotrophic lateral sclerosis (ALS), compared to control samples. A low level of albumin was observed in plasma from ALS patients compared to controls. Additionally, free zone electrophoresis was employed to detect human growth hormone (GH) activity in hormone preparations purified from human pituitaries. We have successfully synthesized antibodies capable of discriminating between dimeric and monomeric GH in samples of clinical origin. To quantify these proteins a calibration curve has been designed, i.e. a plot of the electrophoretic mobility of the complex GH/gel antibody against the protein concentration in the sample, for instance serum or CSF.

    This method was also employed for qualitative and quantitative determinations of Somatropin, a non-glycosylated GH and glycosylated-GH in a body liquid.

    Our results indicate that by this technique one can “fish out” with high accuracy various proteins from both body fluids containing a great number of other proteins. It might well apply also to biomarker proteins for other diseases.

    List of papers
    1. Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples: I. Spectrophotometric approach to design the calibration curve for the quantification
    Open this publication in new window or tab >>Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples: I. Spectrophotometric approach to design the calibration curve for the quantification
    2008 (English)In: Journal of Separation Science, ISSN 1615-9306, E-ISSN 1615-9314, Vol. 31, no 22, p. 3945-3953Article in journal (Refereed) Published
    Abstract [en]

    High selectivity of a biomarker is a basic requirement when it is used for diagnosis, prognosis and treatment of a disease. The artificial gel antibodies, which we synthesise by a molecular imprinting method, have this property not only for proteins, but also for bioparticles, such as viruses and bacteria. However, diagnosis of a disease requires not only that the biomarker can be "fished out" from a body fluid with high selectivity, but also that its concentration in the sample can rapidly be determined and preferably by a simple technique. This paper deals primarily with the development of a spectrophotometric method, which is so simple and fast that it can be used with advantage in a Doctor's Office. The development of this method was not straight-forward. However, by modifications of the performance of these measurements we can now design standard curves in the form of a straight line, when we plot the true (not the recorded "apparent" absorption) against known protein concentrations. In an additional publication (see the following paper in this issue of JSS) we show an application of such a plot: determination of the concentration of albumin in serum and cerebrospinal fluid from patients with neurological disorders to investigate whether albumin is a biomarker for these diseases.

    Keywords
    Apparent absorption, artificial antibodies, CBB, molecular imprinting, true absorption
    National Category
    Pharmaceutical Sciences
    Identifiers
    urn:nbn:se:uu:diva-104856 (URN)10.1002/jssc.200800385 (DOI)000262167500017 ()19065619 (PubMedID)
    Available from: 2009-05-29 Created: 2009-05-29 Last updated: 2018-01-13Bibliographically approved
    2. Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples: II. Albumin in body fluids of patients with neurological disorders
    Open this publication in new window or tab >>Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples: II. Albumin in body fluids of patients with neurological disorders
    2008 (English)In: Journal of Separation Science, ISSN 1615-9306, E-ISSN 1615-9314, Vol. 31, no 22, p. 3954-3958Article in journal (Refereed) Published
    Abstract [en]

    We have previously used the molecular-imprinting method for the synthesis of artificial gel antibodies, highly selective for various proteins. In the present work, we have synthesized artificial gel antibodies against human albumin with the aim to develop a simple and rapid procedure to measure the concentration of this protein in samples of clinical interest. The procedure, based on the design of a standard curve (see the preceding paper), was applied on a quantitative analysis of albumin in human plasma and cerebrospinal fluid (CSF). We found that our technique permitted detection of albumin in these body fluids with high precision and that the concentration of this protein was significantly enhanced in CSF from patients with amyotrophic lateral sclerosis (ALS), compared to control samples. This finding is in agreement with results from earlier studies, which confirms the validity of our analysis technique and suggests that the barrier permeability may be affected in ALS, perhaps also for other proteins. No enhancement in plasma levels of albumin was seen in patients with ALS, but rather a decrease. The results further indicate that our approach might also apply well to other biomarkers for the actual neurological disease and other disorders.

    Keywords
    Amyotrophic lateral sclerosis, artificial gel antibodies, cerebrospinal fluid, human albumin, plasma
    National Category
    Pharmaceutical Sciences
    Identifiers
    urn:nbn:se:uu:diva-104857 (URN)10.1002/jssc.200800386 (DOI)000262167500018 ()19065610 (PubMedID)
    Available from: 2009-05-29 Created: 2009-05-29 Last updated: 2018-01-13Bibliographically approved
    3. Application of artificial gel antibodies for investigating molecular polymorphisms of human pituitary growth hormone
    Open this publication in new window or tab >>Application of artificial gel antibodies for investigating molecular polymorphisms of human pituitary growth hormone
    2011 (English)In: Amino Acids, ISSN 0939-4451, E-ISSN 1438-2199, Vol. 40, no 4, p. 1249-1255Article in journal (Refereed) Published
    Abstract [en]

    Artificial gel-antibodies were used to probe human growth hormone (GH) activity in hormone preparations purified from human pituitaries. A partially purified fraction containing differently sized forms of the hormone was further processed to yield monomeric and dimeric forms of GH activity suitable for synthesizing artificial polyacrylamide gel antibodies. These two types of GH antibodies were used for probing GH activity in experiments analyzing the two forms of the hormone by HPLC gel-permeation and ion-exchange chromatography. In the size-exclusion experiments both hormone fractions eluted as homogenous peaks, whereas the ion exchanger resolved the hormones in several active components. The antibodies towards the GH monomer were more potent to recognize monomeric GH compared to antibodies against dimeric GH. The opposite was found for the dimeric GH antibodies. It was concluded that these two sets of antibodies it might be useful for discriminating between dimeric and monomeric GH in samples of clinical origin.

    Keywords
    Assay, Electrophoresis, Growth hormone (GH), HPLC, Human, Pituitary, Polyacrylamide gel antibodies, Purification, Quantification
    National Category
    Pharmaceutical Sciences
    Research subject
    Pharmaceutical Biochemistry
    Identifiers
    urn:nbn:se:uu:diva-122448 (URN)10.1007/s00726-011-0840-3 (DOI)000288546700021 ()21312046 (PubMedID)
    Available from: 2010-04-16 Created: 2010-04-13 Last updated: 2018-01-12Bibliographically approved
    4. Precautions to improve the accuracy of quantitative determinations of biomarkers in clinical diagnostics
    Open this publication in new window or tab >>Precautions to improve the accuracy of quantitative determinations of biomarkers in clinical diagnostics
    2010 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 31, no 16, p. 2722-2729Article in journal (Refereed) Published
    Abstract [en]

    Although protein biomarkers have a great potential as biomarkers for diagnosis of diseases, they are seldom used in hospitals. There are many reasons for this, for instance, the difficulties to (i) find a biomarker for which the concentration in body fluids clearly differs between patients and healthy subjects, (ii) attain purification of the biomarker close to 100%, which is required for production of conventional protein antibodies as well as artificial gel antibodies for selective capture of a biomarker, (iii) design a standard curve for rapid and accurate determination of the concentration of the biomarker in the body fluid because of adsorption of the biomarker onto vials, pipettes, etc., (iv) determine accurately the sample volume delivered by a pipette, (v) avoid polymerization of the biomarker upon storage and to decide whether it is in the form not only of monomers, but also of dimers, trimers, etc., in the native state, (vi) determine the degree of possible glycosylation and amidation of the biomarker and (vii) decide whether glycosylation and amidation positively or negatively affects the possibility to use the protein as a biomarker. In this article, we discuss in quantitative terms the difficulties (iii-vii) and how to overcome them, which also may help to overcome the difficulty (ti), which in turn minimizes difficulty (i).

    Keywords
    Artificial gel antibodies, Biomarker, Free zone electrophoresis, Growth hormone, Polyacrylamide gels
    National Category
    Chemical Sciences
    Research subject
    Biochemistry
    Identifiers
    urn:nbn:se:uu:diva-122455 (URN)10.1002/elps.201000243 (DOI)000281732900004 ()
    Note
    Uppdaterad från Manuskript till Artikel; 20101202Available from: 2010-04-16 Created: 2010-04-13 Last updated: 2017-12-12Bibliographically approved
  • 107.
    Gising, Johan
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Design and Synthesis of Enzyme Inhibitors Against Infectious Diseases: Targeting Hepatitis C Virus NS3 Protease and Mycobacterium tuberculosis Ribonucleotide Reductase2012Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Infectious diseases, including hepatitis C and tuberculosis, claim the lives of over 15 million people each year. Hepatitis C is caused by the hepatitis C virus (HCV) which infects the liver and can ultimately result in liver transplantation. HCV is very adaptive as a result of its high mutation rate. Thus, there is a potential high risk for the development of drug resistance and also a possible cross-resistance due to a structural similarity between many of the HCV NS3 protease inhibitors currently in clinical trial and on the market, that all are based on a P2-proline or a proline mimic. Thus, part of the research behind this thesis was to explore a new structural P3-P2 unit for the NS3 protease inhibitors, a 2(1H)-pyrazinone moiety. A microwave-assisted protocol was developed, and the 2(1H)-pyrazinone core was synthesized in only 2 × 10 min. A series of optimization steps resulted in several submicromolar 2(1H)-pyrazinone-containing NS3 protease inhibitors that performed well against drug-resistant NS3 protease variants. The key modifications were: exchanging the unstable carbamate P3 capping group for a stable urea functionality, transferring the P2 group from the amino acid residue to the pyrazinone ring and elongating the substituent, and using an aromatic acyl sulfonamide in the P1-P1' position.

    The causative agent of tuberculosis is Mycobacterium tuberculosis (Mtb), which currently infects one third of the world's population. No new TB drugs have been approved in nearly 50 years and drug resistance has been observed for all of the current first-line drugs. Because of the importance of identifying novel drug targets, the ribonucleotide reductase (RNR) enzyme was investigated. The RNR enzyme consists of two R1 and two R2 subunits and is essential for Mtb replication. Starting from hits identified in a virtual screening program, a small library of low molecular weight inhibitors of the association between the R1 and R2 subunits was designed and synthesized. The compounds with the strongest affinity for the R1 subunit of RNR were further evaluated in an orthogonal activity assay. Two RNR inhibitors with promising antimycobacterial effects were identified, which can serve as leads in the further optimization of this class of compounds.

    List of papers
    1. A straightforward microwave method for rapid synthesis of N-1, C-6 functionalized 3,5-dichloro-2(1H)-pyrazinones
    Open this publication in new window or tab >>A straightforward microwave method for rapid synthesis of N-1, C-6 functionalized 3,5-dichloro-2(1H)-pyrazinones
    2009 (English)In: Organic and biomolecular chemistry, ISSN 1477-0520, E-ISSN 1477-0539, Vol. 7, no 13, p. 2809-2815Article in journal (Refereed) Published
    Abstract [en]

    A rapid and versatile one-pot, 2 * 10 min microwave protocol for the prepn. of N-1 and C-6 decorated 3,5-dichloro-2(1H)-pyrazinones, e.g., I, from the corresponding primary amines and aldehyde was developed. Comparable reaction sequences using classical conditions require about 1-2 days of heating. The alpha -aminonitrile was first generated in a Strecker reaction and thereafter cyclized under microwave heating. The microwave approach developed offers the possibility of efficiently generating and utilizing functionalized 3-amino-5-chloro-2(1H)-pyrazinone-N-1-carboxylic acids as beta -strand inducing core structures in a medicinal chem. context. To illustrate the usefulness of the method, the synthesis of two novel 2(1H)-pyrazinone-contg. Hepatitis C virus NS3 protease inhibitors, e.g., II, is reported.

    National Category
    Pharmaceutical Sciences
    Research subject
    Medicinal Chemistry; Organic Chemistry
    Identifiers
    urn:nbn:se:uu:diva-110737 (URN)10.1039/b90550lk (DOI)000267124300020 ()
    Note

    CAN 151:245625 28-17 Heterocyclic Compounds (More Than One Hetero Atom) Journal 1477-0520 written in English. 1178896-10-0P; 1178896-11-1P Role: PAC (Pharmacological activity), SPN (Synthetic preparation), BIOL (Biological study), PREP (Preparation) (prepn. and hepatitis C virus NS3 protease inhibitory activity of (pyrazinylacetamido)cyclopropanoic acid derivs. via coupling reaction, hydrolysis, and peptide coupling of pyrazinylacetate deriv.); 4248-19-5 (tert-Butyl carbamate); 259214-56-7 Role: RCT (Reactant), RACT (Reactant or reagent) (prepn. and hepatitis C virus NS3 protease inhibitory activity of (pyrazinylacetamido)cyclopropanoic acid derivs. via coupling reaction, hydrolysis, and peptide coupling of pyrazinylacetate deriv.); 1178896-08-6P; 1178896-09-7P Role: RCT (Reactant), SPN (Synthetic preparation), PREP (Preparation), RACT (Reactant or reagent) (prepn. and hepatitis C virus NS3 protease inhibitory activity of (pyrazinylacetamido)cyclopropanoic acid derivs. via coupling reaction, hydrolysis, and peptide coupling of pyrazinylacetate deriv.); 62-53-3 (Aniline); 100-46-9 (Benzylamine); 100-52-7 (Benzaldehyde); 108-91-8 (Cyclohexylamine); 109-73-9 (1-Butanamine); 122-78-1 (Benzeneacetaldehyde); 1122-91-4 (4-Bromobenzaldehyde); 2393-23-9 (4-Methoxybenzylamine); 2462-31-9; 3009-97-0; 3010-04-6; 3081-24-1; 3132-99-8 (3-Bromobenzaldehyde); 3182-93-2; 21760-98-5; 37760-98-8 Role: RCT (Reactant), RACT (Reactant or reagent) (prepn. of dichloropyrazinones via Strecker reaction of primary amines, aldehydes, and trimethylsilyl cyanide, followed by cyclocondensation with oxalyl chloride under microwave irradn.); 2182-39-0P; 252271-82-2P; 308845-80-9P; 1178896-02-0P; 1178896-07-5P Role: RCT (Reactant), SPN (Synthetic preparation), PREP (Preparation), RACT (Reactant or reagent) (prepn. of dichloropyrazinones via Strecker reaction of primary amines, aldehydes, and trimethylsilyl cyanide, followed by cyclocondensation with oxalyl chloride under microwave irradn.); 87486-37-1P; 173200-35-6P; 173200-36-7P; 199296-26-9P; 308845-81-0P; 595581-85-4P; 602280-43-3P; 1178895-95-8P; 1178895-96-9P; 1178895-97-0P; 1178895-98-1P; 1178895-99-2P; 1178896-00-8P; 1178896-01-9P; 1178896-03-1P; 1178896-04-2P; 1178896-05-3P; 1178896-06-4P Role: SPN (Synthetic preparation), PREP (Preparation) (prepn. of dichloropyrazinones via Strecker reaction of primary amines, aldehydes, and trimethylsilyl cyanide, followed by cyclocondensation with oxalyl chloride under microwave irradn.)

    Available from: 2009-11-24 Created: 2009-11-24 Last updated: 2018-01-12Bibliographically approved
    2. Discovery of Achiral Inhibitors of the Hepatitis C Virus NS3 Protease based on 2(1H)-pyrazinones
    Open this publication in new window or tab >>Discovery of Achiral Inhibitors of the Hepatitis C Virus NS3 Protease based on 2(1H)-pyrazinones
    Show others...
    2010 (English)In: Bioorganic & Medicinal Chemistry, ISSN 0968-0896, E-ISSN 1464-3391, Vol. 18, no 17, p. 6512-6525Article in journal (Refereed) Published
    Abstract [en]

    Herein, the design, synthesis and inhibitory potency of a series of novel hepatitis C virus (HCV) NS3 protease inhibitors are presented. These inhibitors are based on a 2(1H)-pyrazinone P3 scaffold in combination with either a P2 phenylglycine or a glycine, and they were evaluated on the wild type as well as on two resistant variants of the enzyme, A156T and D168V. Molecular modelling suggested that the aromatic side-chain of the P2 phenylglycine occupies the same space as the substituent in position 6 on the pyrazinone core. The versatile synthetic route applied for the pyrazinone synthesis made a switch between the two positions easily feasible, resulting in phenyl- or benzyl substituted pyrazinones and leaving glycine as the P2 residue. Of several P1-P1′ residues evaluated, an aromatic P1-P1′ scaffold was found superior in combination with the new P3-P2 building block. As a result, an entirely new type of achiral and rigidified inhibitors was discovered, with the best of the novel inhibitors having fourfold improved potency compared to the corresponding tripeptide lead. We consider these achiral inhibitors highly suitable as starting points for further optimization.

    Keywords
    Hepatitis C virus NS3 protease, Protease inhibitors, 2(1H)-pyrazinone, Phenylglycine
    National Category
    Medicinal Chemistry
    Research subject
    Medicinal Chemistry
    Identifiers
    urn:nbn:se:uu:diva-111366 (URN)10.1016/j.bmc.2010.06.101 (DOI)000281203300032 ()20673728 (PubMedID)
    Note

    Uppdaterad från manuskript till Artikel 20101206

    Available from: 2009-12-16 Created: 2009-12-11 Last updated: 2018-01-12Bibliographically approved
    3. Achiral Pyrazinone-Based Inhibitors of the Hepatitis C Virus NS3 Protease and Drug-Resistant Variants with Elongated Substituents Directed Toward the S2 Pocket
    Open this publication in new window or tab >>Achiral Pyrazinone-Based Inhibitors of the Hepatitis C Virus NS3 Protease and Drug-Resistant Variants with Elongated Substituents Directed Toward the S2 Pocket
    Show others...
    2014 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 57, no 5, p. 1790-1801Article in journal (Refereed) Published
    Abstract [en]

    Herein we describe the design, synthesis, inhibitory potency, and pharmacokinetic properties of a novel class of achiral peptidomimetic HCV NS3 protease inhibitors. The compounds are based on a dipeptidomimetic pyrazinone glycine P3P2 building block in combination with an aromatic acyl sulfonamide in the P1P1′ position. Structure–activity relationship data and molecular modeling support occupancy of the S2 pocket from elongated R6 substituents on the 2(1H)-pyrazinone core and several inhibitors with improved inhibitory potency down to Ki = 0.11 μM were identified. A major goal with the design was to produce inhibitors structurally dissimilar to the di- and tripeptide-based HCV protease inhibitors in advanced stages of development for which cross-resistance might be an issue. Therefore, the retained and improved inhibitory potency against the drug-resistant variants A156T, D168V, and R155K further strengthen the potential of this class of inhibitors. A number of the inhibitors were tested in in vitro preclinical profiling assays to evaluate their apparent pharmacokinetic properties. The various R6 substituents were found to have a major influence on solubility, metabolic stability, and cell permeability.

    National Category
    Medicinal Chemistry
    Research subject
    Chemistry with specialization in Organic Chemistry
    Identifiers
    urn:nbn:se:uu:diva-172003 (URN)10.1021/jm301887f (DOI)000333005800011 ()
    Available from: 2012-03-31 Created: 2012-03-31 Last updated: 2018-01-12Bibliographically approved
    4. Design and synthesis of ribonucleotide reductase inhibitors with activity against Mycobacterium tuberculosis
    Open this publication in new window or tab >>Design and synthesis of ribonucleotide reductase inhibitors with activity against Mycobacterium tuberculosis
    Show others...
    (English)Manuscript (preprint) (Other academic)
    National Category
    Medicinal Chemistry
    Research subject
    Medicinal Chemistry
    Identifiers
    urn:nbn:se:uu:diva-172340 (URN)
    Available from: 2012-04-04 Created: 2012-04-04 Last updated: 2018-01-12
  • 108.
    Gising, Johan
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Belfrage, Anna Karin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Alogheli, Hiba
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Ehrenberg, Angelica
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC.
    Åkerblom, Eva
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Svensson, Richard
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Artursson, Per
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Anders, Karlén
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Danielsson, U. Helena
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC.
    Larhed, Mats
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Sandström, Anja
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Achiral Pyrazinone-Based Inhibitors of the Hepatitis C Virus NS3 Protease and Drug-Resistant Variants with Elongated Substituents Directed Toward the S2 Pocket2014In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 57, no 5, p. 1790-1801Article in journal (Refereed)
    Abstract [en]

    Herein we describe the design, synthesis, inhibitory potency, and pharmacokinetic properties of a novel class of achiral peptidomimetic HCV NS3 protease inhibitors. The compounds are based on a dipeptidomimetic pyrazinone glycine P3P2 building block in combination with an aromatic acyl sulfonamide in the P1P1′ position. Structure–activity relationship data and molecular modeling support occupancy of the S2 pocket from elongated R6 substituents on the 2(1H)-pyrazinone core and several inhibitors with improved inhibitory potency down to Ki = 0.11 μM were identified. A major goal with the design was to produce inhibitors structurally dissimilar to the di- and tripeptide-based HCV protease inhibitors in advanced stages of development for which cross-resistance might be an issue. Therefore, the retained and improved inhibitory potency against the drug-resistant variants A156T, D168V, and R155K further strengthen the potential of this class of inhibitors. A number of the inhibitors were tested in in vitro preclinical profiling assays to evaluate their apparent pharmacokinetic properties. The various R6 substituents were found to have a major influence on solubility, metabolic stability, and cell permeability.

  • 109.
    Gising, Johan
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Belfrage, Anna Karin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Örtqvist, Pernilla
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Larhed, Mats
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Danielson, U. Helena
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Sandström, Anja
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry.
    Hepatitis C protease inhibitors based on 2(1H)-pyrazinones2010In: Abstract of Papers of the American Chemical Society, ISSN 0065-7727, Vol. 240, p. 116-MEDI-Article in journal (Refereed)
  • 110.
    Gising, Johan
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Jansson, Anna
    Daniel, Ericsson
    Hiba, Alogheli
    Larhed, Mats
    Unge, Torsten
    Karlén, Anders
    Design and synthesis of ribonucleotide reductase inhibitors with activity against Mycobacterium tuberculosisManuscript (preprint) (Other academic)
  • 111.
    Graffner Nordberg, Malin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Approaches to soft drug analogues of dihydrofolate reductase inhibitors: Design and synthesis2001Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The main objective of the research described in this thesis has been the design and synthesis of inhibitors of the enzyme dihydrofolate reductase (DHFR) intended for local administration and devoid of systemic side-effects. The blocking of the enzymatic activity of DHFR is a key element in the treatment of many diseases, including cancer, bacterial and protozoal infections, and also opportunistic infections associated with AIDS (Pneumocystis carinii pneumonia, PCP). Recent research indicates that the enzyme also is involved in various autoimmune diseases, e.g., rheumatoid arthritis, inflammatory bowel diseases and psoriasis. Many useful antifolates have been developed to date although problems remain with toxicity and selectivity, e.g., the well-established, classical antifolate methotrexate exerts a high activity but also high toxicity. The new antifolates described herein were designed to retain the pharmacophore of methotrexate, but encompassing an ester group, so that they also would serve as substrates for the endogenous hydrolytic enzymes, e.g., esterases. Such antifolates would optimally comprise good examples of soft drugs because they in a controlled fashion would be rapidly and predictably metabolized to non-toxic metabolites after having exerted their biological effect at the site of administration.

    A preliminary screening of a large series of simpler aromatic esters as model compounds in a biological assay consisting of esterases from different sources was performed. The structural features of the least reactive ester were substituted for the methyleneamino bridge in methotrexate to produce analogues that were chemically stable but potential substrates for DHFR as well as for the esterases.

    The new inhibitor showed desirable activity towards rat liver DHFR, being only eight times less potent then methotrexate. Furthermore, the derived metabolites were found to be poor substrates for the same enzyme. The new compound showed good activity in a mice colitis model in vivo, but a pharmacokinetic study revealed that the half-life of the new compound was similar to methotrexate. A series of compounds characterized by a high lipophilicity and thus expected to provide better esterase substrates were designed and synthesized. One of these analogues in which three methoxy groups were substituted for the glutamic residue of methotrexate exhibited favorable pharmacokinetics. This compound is structurally similar to another potent DHFR inhibitor, trimetrexate, used in the therapy of PCP (vide supra). The new inhibitor that undergoes a fast metabolism in vivo is suitable as a model to further investigate the soft drug concept.

  • 112. Guimond, Marie-Odile
    et al.
    Hallberg, Mathias
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
    Gallo-Payet, Nicole
    Wallinder, Charlotta
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Saralasin and Sarile Are AT2 Receptor Agonists2014In: ACS Medicinal Chemistry Letters, ISSN 1948-5875, E-ISSN 1948-5875, Vol. 5, no 10, p. 1129-1132Article in journal (Refereed)
    Abstract [en]

    Saralasin and sarile, extensively studied over the past 40 years as angiotensin II (Ang II) receptor blockers, induce neurite outgrowth in a NG108-15 cell assay to a similar extent as the endogenous Ang II. In their undifferentiated state, these cells express mainly the AT2 receptor. The neurite outgrowth was inhibited by preincubation with the AT2 receptor selective antagonist PD 123,319, which suggests that the observed outgrowth was mediated by the AT2 receptor. Neither saralasin nor sarile reduced the neurite outgrowth induced by Ang II proving that the two octapeptides do not act as antagonists at the AT2 receptor and may be considered as AT2 receptor agonists.

  • 113.
    Gustafsson, Simon
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Nanotechnology and Functional Materials.
    Mihranyan, Albert
    Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Nanotechnology and Functional Materials.
    Investigating Protein Throughput, Vmax Values and Virus Removal Efficiency of the Paper Based Mille-Feuille Filter2017In: Abstract of Papers of the American Chemical Society, ISSN 0065-7727, Vol. 253Article in journal (Other academic)
    Abstract [en]

    In this work we present Vmax values and throughput for the nanocellulose-based Mille-feuille virus removal filter paper as well as evaluate the affinity of different proteins for the cellulose surface using quartz crystal microbalance (QCMB). The Minute Virus of Mice (MVM) virus removal result for the Mille-feuille filter is also presented. The nanocellulose-based Mille-feuille filter is a non-woven, µm-thick filter paper with a narrow and tailorable pore size distribution, highly suitable for removing viruses of all sizes, including the worst case model Minute Virus of Mice (MVM). It is produced by hot-pressing method rather than by phase-inversion which is otherwise used to manufacture industrial analogues. Robust and cost-efficient virus removal processes are in high demand for the biotechnology industry, especially for the production of monoclonal antibodies derived from mammalian cell lines, therapeutic proteins derived from human plasma, and cell culture media. Non-woven filters have the advantage in contrast to regular phase-inversion made filters of a much higher porosity. For instance, the Mille-feuille filter has a porosity of 42% compared to 10-15% of the functional skin layer in asymmetric virus removal filters used today. In theory a higher porosity will result in higher flow rates. However, it also means that there is a larger filter area exposed to filtrate. Therefore investigating the surface affinity for proteins with different hydrophobicity and charge using QCMB is of great importance to be able to predict protein filtration behaviour. The presented work is a part of continuous efforts to develop a new class of highly efficient and affordable virus removal filters.

  • 114. Gybaeck, Helena
    et al.
    Haeberlein, Markus
    Jonasson, Catrin
    Kihlstroem, Jacob
    Molin, Haakan
    Plobeck, Niklas
    Svensson, Andreas
    Viklund, Jenny
    Yngve, Ulrika.
    Benzoic acid derivatives as glycine receptor inhibitors, their preparation, pharmaceutical compositions, and use in therapy.2006Patent (Other (popular science, discussion, etc.))
    Abstract [en]

    The invention relates to benzoic acid derivs. of formula I, which are inhibitors of glycine receptors (GlyRs). In compds. I, Y is H, OH, halo, (un)substituted C1-6 alkoxy, or (un)substituted C1-6 alkyl; R1 is (un)substituted C3-6 cycloalkyl, (un)substituted heterocyclyl, (un)substituted aryl, (un)substituted alkylaryl, (un)substituted heteroaryl, or (un)substituted C3-6 alkyl; L is selected from a bond, heteroarylene, -C(O)-, -CH2-, -CH(OR4)-, -N(OH)-, -N(R4)-, -S(O)n-, where R4 is H or C1-6 alkyl and n is 0, 1, or 2; R2 is H, halo, cyano, dimethylamino, (un)substituted C1-6 alkyl, (un)substituted C3-6 cycloalkyl, (un)substituted heterocyclyl, (un)substituted aryl, (un)substituted alkylaryl, (un)substituted heterocyclyl or (un)substituted heteroaryl; and R3 is OH or C1-6 alkoxy; with the exclusion of several compds. The invention also relates to the prepn. of I, pharmaceutical compns. comprising a therapeutically effective amt. of a compd. I with one or more pharmaceutically acceptable diluents, excipients and/or inert carriers, as well as to the use of the compns. for the treatment of conditions that respond to glycine receptor inhibition, such as neuropathic or inflammatory pain syndromes. Chlorination of 4-methoxybenzenethiol followed by C-substitution with 3-tert-butyl-2-hydroxy-6-methylbenzoic acid and oxidn. gave (arylsulfonyl)benzoic acid II. Several compds. of the invention expressed IC50 values against human GlyR α1 of about 10 nM to about 30 μM (no specific data). [on SciFinder(R)]

  • 115. Gybaeck, Helena
    et al.
    Haeberlein, Markus
    Jonasson, Catrin
    Kihlstroem, Jacob
    Molin, Haakan
    Plobeck, Niklas
    Svensson, Andreas
    Viklund, Jenny
    Yngve, Ulrika.
    Benzoic acid derivatives as glycine receptor inhibitors, their preparation, pharmaceutical compositions, and use in therapy.2006Patent (Other (popular science, discussion, etc.))
    Abstract [en]

    The invention relates to benzoic acid derivs. of formula I, which are inhibitors of glycine receptors (GlyRs). In compds. I, Y is H, OH, halo, (un)substituted C1-6 alkoxy, or (un)substituted C1-6 alkyl; R1 is (un)substituted C3-6 cycloalkyl, (un)substituted heterocyclyl, (un)substituted aryl, (un)substituted alkylaryl, (un)substituted heteroaryl, or (un)substituted C3-6 alkyl; L is selected from a bond, heteroarylene, -C(O)-, -CH2-, -CH(OR4)-, -N(OH)-, -N(R4)-, -S(O)n-, where R4 is H or C1-6 alkyl and n is 0, 1, or 2; R2 is H, halo, cyano, dimethylamino, (un)substituted C1-6 alkyl, (un)substituted C3-6 cycloalkyl, (un)substituted heterocyclyl, (un)substituted aryl, (un)substituted alkylaryl, (un)substituted heterocyclyl or (un)substituted heteroaryl; and R3 is OH or C1-6 alkoxy; with the exclusion of several compds. The invention also relates to the prepn. of I, pharmaceutical compns. comprising a therapeutically effective amt. of a compd. I with one or more pharmaceutically acceptable diluents, excipients and/or inert carriers, as well as to the use of the compns. for the treatment of conditions that respond to glycine receptor inhibition, such as neuropathic or inflammatory pain syndromes. Chlorination of 4-methoxybenzenethiol followed by C-substitution with 3-tert-butyl-2-hydroxy-6-methylbenzoic acid and oxidn. gave (arylsulfonyl)benzoic acid II. Several compds. of the invention expressed IC50 values against human GlyR α1 of about 10 nM to about 30 μM (no specific data). [on SciFinder(R)]

  • 116.
    Gyllenhaal, Olle
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Karlsson, Anders
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Evaluation Conditions for SFC of Metoprolol and Related Amino Alcohols on Hypercarb (Porous Graphitic Carbon) with Respect to Structure–Selectivity Relations2010In: Chromatographia, ISSN 0009-5893, E-ISSN 1612-1112, Vol. 71, no 1-2, p. 7-13Article in journal (Refereed)
    Abstract [en]

    Mobile phase chromatographic conditions for the selective separation of metoprolol from related amino alcohols have been evaluated using Hypercarb as support and carbon dioxide with addition of methanol as mobile phase. The objective for the presented study was to show the unique ability of the porous graphitized carbon surface to separate closely structurally related substances. Experimental results, using Hypercarb and “chromatographic normal phase conditions” (SFC), are presented with focus on how to control retention and to improve peak performance. A high concentration of basic aliphatic amine additive was required in order to elute the amine analytes as symmetrical peaks. N,N-Dimethyloctylamine was preferred over triethylamine since the retention was markedly shorter though the selectivity and resolution were virtually the same. The selectivity can be optimized by altering the temperature of the column. A high selectivity was demonstrated between metoprolol and two homologues with one and two extra methylene groups inserted between the secondary nitrogen atom and the carbon atom with a hydroxyl group attached. The mobile phase flow rate effect the column efficiency only to a minor extent as the steepness of the van Deemter curve for metoprolol was virtually flat in the range studied: 0.5–3.0 mL min−1.

  • 117.
    Göransson, Ulf
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Macrocyclic polypeptides from plants2002Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The aim of this work was to explore the structural and functional diversity of polypeptides that are found in plants. Expanding knowledge of simililarities between plant use of these compound and animal use promises exceptional opportunities for finding, from plant research, new structures with biomedical and biotechnological potential.

    A fractionation protocol was developed and applied to many plant species, providing fractions enriched in polypeptides, amenable to chemical and biological evaluation. From one species, the common field pansy (Viola arvensis), a 29-amino-acid residue polypeptide was isolated, named varv A, which revealed a remarkable macrocyclic structure (i.e., N- and C-termini are joined) stabilised by three knotted disulfides.

    Varv A, together with an increasing number of homologous peptides, form the currently known peptide family of cyclotides. Their stable structure makes them an attractive scaffold for protein engineering. In addition, they display a wide range of biological activities (e.g., antimicrobial, cytotoxic, and insecticidal). As a part of this work, the cytotoxic effects of varv A and two other isolated cyclotides were evaluated in a human cell-line panel: all were active in the low µM range. Most likely, these effects involve pore formation through cell membranes.

    Cyclotides were found to be common in the plant family Violaceae; with eleven cyclotides isolated and sequenced from V. arvensis, V. cotyledon, and Hybanthus parviflorus. For six members of the genus Viola, cyclotide expression profiles were examined by liquid chromatography-mass spectrometry (LC-MS): all expressed notably complex mixtures, with single species containing more than 50 cyclotides. These profiles reflect the evolution of the genus.

    To assess these mixtures, a rational strategy for MS based amino acid sequencing of cyclotides was developed, circumventing inherent structural problems, such as low content of positively charged amino acids and the macrocyclic structure. This was achieved by aminoethylation of cysteines, which, following tryptic digestion, produced fragments of size and charge amenable to MS analysis. This method was also modified and used for mapping of disulfide bonds.

    Methods for isolation and characterisation developed in this work may prove useful not only for further studies on macrocyclic polypeptides from plants, but also for other plant peptides and disulfide-rich peptides from animals.

    List of papers
    1. Fractionation protocol for the isolation of polypeptides from plant biomass
    Open this publication in new window or tab >>Fractionation protocol for the isolation of polypeptides from plant biomass
    Show others...
    1998 (English)In: Journal of natural products (Print), ISSN 0163-3864, E-ISSN 1520-6025, Vol. 61, no 1, p. 77-81Article in journal (Refereed) Published
    Abstract [en]

    A fractionation protocol for the isolation of a highly purified polypeptide fraction from plant biomass is described. The procedure dereplicates ubiquitous substance classes known to interfere with bioassays often used in natural product-based drug discovery programs. The protocol involves pre-extraction with dichloromethane, extraction with ethanol (50%), removal of tannins with polyamide, removal of low-molecular-weight components with size-exclusion chromatography over Sephadex G-10, and final removal of salts and polysaccharides with solid-phase extraction using reversed-phase cartridges. The method has been applied to the aerial parts of Viola arvensis, resulting in the isolation of a peptide fraction that on further separation yielded a novel 29-residue macrocyclic polypeptide named varv peptide A, cyclo(-TCVGGTCNTPGCSCSWPVCTRNGLPVCGE-).

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-89791 (URN)10.1021/np970342r (DOI)9548831 (PubMedID)
    Available from: 2002-04-05 Created: 2002-04-05 Last updated: 2017-12-14Bibliographically approved
    2. Seven novel macrocyclic polypeptides from Viola arvensis
    Open this publication in new window or tab >>Seven novel macrocyclic polypeptides from Viola arvensis
    Show others...
    1999 In: J Nat Prod, Vol. 62, no 2, p. 283-286Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-89792 (URN)
    Available from: 2002-04-05 Created: 2002-04-05Bibliographically approved
    3. First cyclotide from Hybanthus (Violaceae)
    Open this publication in new window or tab >>First cyclotide from Hybanthus (Violaceae)
    Show others...
    2001 (English)In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 58, no 1, p. 47-51Article in journal (Refereed) Published
    Abstract [en]

    Hypa A, a novel macrocyclic polypeptide containing 30 amino acid residues, has been isolated from the n-butanol extract of the Argentine plant Hybanthus parviflorus. The sequence, cyclo-(SCVYIPCTITALLGCSCKNKVCYNGIPCAE), was determined by automated Edman degradation, quantitative amino acid analysis and nanospray MS/MS(2). Three intramolecular disulfide bridges stabilize the cyclic peptide backbone of hypa A. Using these structural features to classify the peptide as a cyclotide, we extended the distribution of that substance class to a new genus, and now propose a uniform nomenclature for cyclotides.

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-89793 (URN)10.1016/S0031-9422(01)00173-X (DOI)11524112 (PubMedID)
    Available from: 2002-04-05 Created: 2002-04-05 Last updated: 2017-12-14Bibliographically approved
    4. Cyclotides: a novel type of cytotoxic agents
    Open this publication in new window or tab >>Cyclotides: a novel type of cytotoxic agents
    Show others...
    2002 (English)In: Molecular Cancer Therapeutics, ISSN 1535-7163, E-ISSN 1538-8514, Vol. 1, no 6, p. 365-369Article in journal (Refereed) Published
    Abstract [en]

    Cytotoxic activities of three naturally occurring macrocyclic peptides (cyclotides) isolated from the two violets, Viola arvensis Murr. and Viola odorata L., were investigated. A nonclonogenic fluorometric microculture assay was used to examine cytotoxicity in a panel of 10 human tumor cell lines representing defined types of cytotoxic drug resistance. Additionally, primary cultures of tumor cells from patients, and for comparison normal lymphocytes, were used to quantify cytotoxic activity. All three cyclotides, varv A, varv F, and cycloviolacin O2, exhibited strong cytotoxic activities, which varied in a dose-dependent manner. Cycloviolacin O2 was the most potent in all cell lines (IC50 0.1– 0.3 _M), followed by varv A (IC50 2.7–6.35 _M) and varv F (IC50 2.6 –7.4 _M), respectively. Activity profiles of the cyclotides differed significantly from those of antitumor drugs in clinical use, which may indicate a new mode of action. This, together with the exceptional chemical and biological stability of cyclotides, makes them interesting in particular for their potential as pharmacological tools and possibly as leads to antitumor agents.

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-92805 (URN)000178770300001 ()12477048 (PubMedID)
    Note

    De två första författarna delar på förstaförfattarskapet.

    Available from: 2005-04-01 Created: 2005-04-01 Last updated: 2017-12-14Bibliographically approved
    5. Peptide profiling by LC-MS and MS sequencing of intercysteine loops: Expression profiles of Viola cyclotides
    Open this publication in new window or tab >>Peptide profiling by LC-MS and MS sequencing of intercysteine loops: Expression profiles of Viola cyclotides
    Article in journal (Refereed) Submitted
    Identifiers
    urn:nbn:se:uu:diva-89795 (URN)
    Available from: 2002-04-05 Created: 2002-04-05Bibliographically approved
  • 118.
    Göransson, Ulf
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
    Burman, Robert
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
    Gunasekera, Sunithi
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
    Strömstedt, Adam A.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
    Rosengren, K. Johan
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
    Circular Proteins from Plants and Fungi2012In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 287, no 32, p. 27001-27006Article, review/survey (Refereed)
    Abstract [en]

    Circular proteins, defined as head-to-tail cyclized polypeptides originating from ribosomal synthesis, represent a novel class of natural products attracting increasing interest. From a scientific point of view, these compounds raise questions of where and why they occur in nature and how they are formed. From a rational point of view, these proteins and their structural concept may be exploited for crop protection and novel pharmaceuticals. Here, we review the current knowledge of three protein families: cyclotides and circular sunflower trypsin inhibitors from the kingdom of plants and the Amanita toxins from fungi. A particular emphasis is placed on their biological origin, structure, and activity. In addition, the opportunity for discovery of novel circular proteins and recent insights into their mechanism of action are discussed.

  • 119.
    Götmar, Gustaf
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Heterogeneous Adsorption in Chiral LC2002Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The adsorption of chiral drugs on proteins immobilized on silica shows an heterogeneous adsorption behavior on multiple sites. The theory of nonlinear chromatography was used for a systematic study on this behavior - linear chromatography can only measure the "global" retention.

    Frontal analysis in the staircase mode was used for studies on the adsorption of model compounds on two different protein columns; β-blockers to cellobiohydrolase I and analytes of different protolytic classes to α1-acid glycoprotein. It was found that the compounds were retained at different kinds of sites, nonselective and enantioselective, and the adsorption was best fitted by the bi-Langmuir model.

    With increasing the eluent pH the interaction of both enantiomers of the amines to both types of sites was increased on α1-acid glycoprotein. In the case of cellobiohydrolase I (Cel7A) only the interaction of the second eluted enantiomer increased, resulting in a dramatic increase of the enantioselectivity with increasing pH. When increasing the eluent pH the linear retention of the second eluted enantiomer switches from exothermic to endothermic behavior on Cel7A, this point depends on the relative retention on endothermic enantioselective sites. The hydrophobicity of the compound was found to be an important property regarding retention behavior for both enantiomers and on both types of sites on Cel7A. A very interesting feature was observed: 2-phenylbutyric acid has a maximum of retention at an intermediate eluent pH on α1-acid glycoprotein and from nonlinear studies it was shown that this originates from maximum interaction strength at the enantioselective sites. For both columns, there is a 1:1 stochiometry at the enantioselective site.

    Protein columns sometimes shows slow and heterogeneous kinetics, and simulation studies were performed to illustrate the consequences of such a behavior. The relations between (i) the coefficient of kinetics (ii) the column length and (iii) the linear flow rate were investigated. A conclusion for the homogeneous case, is that the solution of all skewed peaks is perfectly Gaussian, if the column hold-up time is high enough.

    List of papers
    1. Apparent and True Enantioselectivity in Enantioseparations
    Open this publication in new window or tab >>Apparent and True Enantioselectivity in Enantioseparations
    2000 In: Chirality, Vol. 12, no 7, p. 558-564Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-89665 (URN)
    Available from: 2002-02-25 Created: 2002-02-25Bibliographically approved
    2. Dependence on the Mobile-Phase pH of the Adsorption Behavior of Propranolol Enantiomers on a Cellulase Protein Used as the Chiral Selector
    Open this publication in new window or tab >>Dependence on the Mobile-Phase pH of the Adsorption Behavior of Propranolol Enantiomers on a Cellulase Protein Used as the Chiral Selector
    1999 In: J. Am. Chem. Soc., Vol. 121, no 6, p. 1164-1174Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-89666 (URN)
    Available from: 2002-02-25 Created: 2002-02-25Bibliographically approved
    3. Retention Mechanism of β-blockers on an Immobilized Cellulase. Relative Importance of the Hydrophobic and Ionic Contributions to Their Enantioselective and Nonselective Interactions
    Open this publication in new window or tab >>Retention Mechanism of β-blockers on an Immobilized Cellulase. Relative Importance of the Hydrophobic and Ionic Contributions to Their Enantioselective and Nonselective Interactions
    2000 In: Anal. Chem, Vol. 72, no 16, p. 3908-3915Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-89667 (URN)
    Available from: 2002-02-25 Created: 2002-02-25Bibliographically approved
    4. Influence of the Solute Hydrophobicity on the Enantioselective Adsorption of β-blockers on a Cellulase Protein Used as the Chiral Selector
    Open this publication in new window or tab >>Influence of the Solute Hydrophobicity on the Enantioselective Adsorption of β-blockers on a Cellulase Protein Used as the Chiral Selector
    2001 In: J. Chromatogr. A, Vol. 905, no 1-2, p. 3-17Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-89668 (URN)
    Available from: 2002-02-25 Created: 2002-02-25Bibliographically approved
    5. Investigation of the Heterogeneous Adsorption Behavior of Selected Enantiomers on Immobilized α1 - Acid Glycoprotein (CHIRAL-AGP®)
    Open this publication in new window or tab >>Investigation of the Heterogeneous Adsorption Behavior of Selected Enantiomers on Immobilized α1 - Acid Glycoprotein (CHIRAL-AGP®)
    In: Anal. Chem.Article in journal (Refereed) Submitted
    Identifiers
    urn:nbn:se:uu:diva-89669 (URN)
    Available from: 2002-02-25 Created: 2002-02-25Bibliographically approved
    6. Peak Tailing and Mass Transfer Kinetics in Linear Chromatography - Dependence on the Column Length and the Linear Velocity of the Mobile Phase
    Open this publication in new window or tab >>Peak Tailing and Mass Transfer Kinetics in Linear Chromatography - Dependence on the Column Length and the Linear Velocity of the Mobile Phase
    1999 In: J. Chromatogr. A, Vol. 831, no 1, p. 17-35Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-89670 (URN)
    Available from: 2002-02-25 Created: 2002-02-25Bibliographically approved
  • 120.
    Haglöf, Jakob
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Enantiomeric Separations using Chiral Counter-Ions2010Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    This thesis describes the use of chiral counter-ions for the enantiomeric separation of amines in non-aqueous capillary electrophoresis. The investigations have been concentrated on studies of the influence, of the chiral counter-ion, the solvent, the electrolyte and the analyte, on the enantioselective separation.

    Modified divalent dipeptides have been introduced in capillary electrophoresis for the separation of amino alcohols and chiral resolution of amines. Association constants for the ion-pair between dipeptide and amino alcohol could be utilized for development of separation systems with higher amino alcohol selectivity. Chiral discrimination (ion-pair formation) between the dipeptides and amines are preferably generated in non-aqueous background electrolytes (BGEs). The amount of triethylamine in the BGE determined the dipeptide charge and a divalent dipeptide promoted higher enantioselectivity than a monovalent dipeptide. An N-terminal-end blocking group and glutamic acid at the C-terminal-end of the dipeptide was important for chiral separation of the amines.

    Chemometric and univariate methods have been employed for evaluation of suitable solvent compositions in the BGE. An experimental design including a single solvent as well as binary, ternary and quaternary mixtures of polar organic solvents, showed that optimal enantioresolution was obtained with an ethanol:methanol 80:20 mixture in the BGE.  Furthermore, water was found to have an adverse influence on enantioselectivity and no enantioresolution was obtained with BGEs containing more than 30 % water.

    An alkali metal hydroxide added to the BGE affected the chiral separation by competing ion-pair formation with the selector. The electroosmosis was reduced in order of decreasing alkali metal ion solvated radius and became anodic using K, Rb or Cs in ethanolic BGEs.

    The correlation between the amino alcohol structure and the enantioselectivity was investigated using chemometrics. The obtained models showed that enantioselectivity for the amino alcohols was promoted by e.g. degree of substitution and substituent size on the nitrogen.

    List of papers
    1.
    The record could not be found. The reason may be that the record is no longer available or you may have typed in a wrong id in the address field.
    2. Separation of amino alcohols using divalent dipeptides as counter ions in aqueous CE
    Open this publication in new window or tab >>Separation of amino alcohols using divalent dipeptides as counter ions in aqueous CE
    2010 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 31, no 10, p. 1706-1712Article in journal (Refereed) Published
    Abstract [en]

    Divalent dipeptides have been introduced as counter ions in aqueous CZE. The dipeptides form ion pairs with amino alcohols in the BGE and facilitate the separation of amino alcohols. High concentrations of dipeptide caused reversed effective mobility for the analytes. The net charge of the dipeptide can be controlled using a buffer or a strong base, and regulates the interaction between the dipeptide and the amino alcohol. A stronger interaction and higher selectivity of amino alcohols was observed when the dipeptides were used as divalent counter ions, than in monovalent or uncharged form. Association constants for ion pairs between divalent dipeptides and amino alcohols can be used to enhance selectivity for amino alcohols in CZE. No chiral separation of amino alcohols was observed when using the dipeptides as ion-pairing chiral selectors in aqueous BGE, but addition of methanol to the BGE promoted enantioselectivity.

    Keywords
    Association constants, CE, divalent counter ion, Ion-pair formation, reversed mobility
    National Category
    Medicinal Chemistry
    Research subject
    Analytical Pharmaceutical Chemistry
    Identifiers
    urn:nbn:se:uu:diva-125470 (URN)10.1002/elps.200900702 (DOI)000278650500015 ()20414882 (PubMedID)
    Available from: 2010-05-19 Created: 2010-05-19 Last updated: 2018-01-12Bibliographically approved
    3. Enantiomeric separation of amines using divalent dipeptides as chiral counter-ions in non-aqueous CE
    Open this publication in new window or tab >>Enantiomeric separation of amines using divalent dipeptides as chiral counter-ions in non-aqueous CE
    (English)Manuscript (preprint) (Other academic)
    Keywords
    Capillary electrophoresis, nonaqueous solvents, divalent chiral counter ion, ion-pairing
    National Category
    Medicinal Chemistry
    Research subject
    Analytical Pharmaceutical Chemistry
    Identifiers
    urn:nbn:se:uu:diva-130047 (URN)
    Available from: 2010-08-30 Created: 2010-08-30 Last updated: 2018-01-12
    4. Multivariate data analysis of a chiral separation system using N-derivatized dipeptides as chiral counter-ions on porous graphitic carbon stationary phase
    Open this publication in new window or tab >>Multivariate data analysis of a chiral separation system using N-derivatized dipeptides as chiral counter-ions on porous graphitic carbon stationary phase
    (English)Manuscript (preprint) (Other academic)
    Keywords
    Chiral counter ion, selectivity, molecular descriptor, topological index, PCA, PLS, OPLS
    National Category
    Medicinal Chemistry
    Research subject
    Analytical Pharmaceutical Chemistry
    Identifiers
    urn:nbn:se:uu:diva-130048 (URN)
    Available from: 2010-08-30 Created: 2010-08-30 Last updated: 2018-01-12
  • 121.
    Haglöf, Jakob
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Karlsson, Anders
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Multivariate data analysis of a chiral separation system using N-derivatized dipeptides as chiral counter-ions on porous graphitic carbon stationary phaseManuscript (preprint) (Other academic)
  • 122.
    Haglöf, Jakob
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Pettersson, Curt
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Enantiomeric separation of amines using divalent dipeptides as chiral counter-ions in non-aqueous CEManuscript (preprint) (Other academic)
  • 123.
    Haglöf, Jakob
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Pettersson, Curt
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Separation of amino alcohols using divalent dipeptides as counter ions in aqueous CE2010In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 31, no 10, p. 1706-1712Article in journal (Refereed)
    Abstract [en]

    Divalent dipeptides have been introduced as counter ions in aqueous CZE. The dipeptides form ion pairs with amino alcohols in the BGE and facilitate the separation of amino alcohols. High concentrations of dipeptide caused reversed effective mobility for the analytes. The net charge of the dipeptide can be controlled using a buffer or a strong base, and regulates the interaction between the dipeptide and the amino alcohol. A stronger interaction and higher selectivity of amino alcohols was observed when the dipeptides were used as divalent counter ions, than in monovalent or uncharged form. Association constants for ion pairs between divalent dipeptides and amino alcohols can be used to enhance selectivity for amino alcohols in CZE. No chiral separation of amino alcohols was observed when using the dipeptides as ion-pairing chiral selectors in aqueous BGE, but addition of methanol to the BGE promoted enantioselectivity.

  • 124. Hajrezaie, Maryam
    et al.
    Salehen, NurAin
    Karimian, Hamed
    Zahedifard, Maryam
    Shams, Keivan
    Al Batran, Rami
    Majid, Nazia Abdul
    Khalifa, Shaden A. M.
    Ali, Hapipah Mohd
    El-Seedi, Hesham
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
    Abdulla, Mahmood Ameen
    Biochanin A Gastroprotective Effects in Ethanol-Induced Gastric Mucosal Ulceration in Rats2015In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 10, no 3, article id e0121529Article in journal (Refereed)
    Abstract [en]

    Background Biochanin A notable bioactive compound which is found in so many traditional medicinal plant. In vivo study was conducted to assess the protective effect of biochanin A on the gastric wall of Spraguedawley rats' stomachs. Methodology The experimental set included different animal groups. Specifically, four groups with gastric mucosal lesions were receiving either a) Ulcer control group treated with absolute ethanol (5 ml/kg), b) 20 mg/kg of omeprazole as reference group, c) 25 of biochanin A, d) 50 mg/kg of biochanin A. Histopathological sectioning followed by immunohistochemistry staining were undertaken to evaluate the influence of the different treatments on gastric wall mucosal layer. The gastric secretions were collected in the form of homogenate and exposed to superoxide dismutase (SOD) and nitric oxide enzyme (NO) and the level of malondialdehyde (MDA) and protein content were measured. Ulceration and patchy haemorrhage were clearly observed by light microscopy. The morphology of the gastric wall as confirmed by immunohistochemistry and fluorescent microscopic observations, exhibited sever deformity with notable thickness, oedematous and complete loss of the mucosal coverage however the biochanin-pretreated animals, similar to the omeprazole-pretreated animals, showed less damage compared to the ulcer control group. Moreover, up-regulation of Hsp70 protein and down-regulation of Bax protein were detected in the biochanin A pre-treated groups and the gastric glandular mucosa was positively stained with Periodic Acid Schiff (PAS) staining and the Leucocytes infiltration was commonly seen. Biochanin A displayed a great increase in SOD and NO levels and decreased the release of MDA. Conclusions This gastroprotective effect of biochanin A could be attributed to the enhancement of cellular metabolic cycles perceived as an increase in the SOD, NO activity, and decrease in the level of MDA, and also decrease in level of Bax expression and increase the Hsp70 expression level.

  • 125.
    Hall, Håkan
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Preclinical PET Platform.
    Velikyan, Irina
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Radiology, Oncology and Radiation Science, Biomedical Radiation Sciences.
    Blom, Elisabeth
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry.
    Ulin, Johan
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry.
    Monazzam, Azita
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Radiology, Oncology and Radiation Science, Biomedical Radiation Sciences.
    Påhlman, Lars
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences, Colorectal Surgery.
    Micke, Patrick
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular and Morphological Pathology.
    Wanders, Alkwin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular and Morphological Pathology.
    McBride, William
    Goldenberg, David M
    Långström, Bengt
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry.
    In vitro autoradiography of carcinoembryonic antigen in tissue from patients with colorectal cancer using multifunctional antibody TF2 and 67/68Ga-labeled haptens by pretargeting2012In: American journal of nuclear medicine and molecular imaging, ISSN 2160-8407, Vol. 2, no 2, p. 141-150Article in journal (Refereed)
    Abstract [en]

    The carcinoembryonic antigen (CEA) was visualized in vitro in tissue from patients with colorectal cancer with trivalent bispecific antibody TF2 and two hapten molecules, [67/68Ga]Ga-IMP461 and [67/68Ga]Ga-IMP485 by means of pretargeting. Colorectal cancer tissue samples obtained from surgery at Uppsala University Hospital, were frozen fresh and cryosectioned. The two hapten molecules comprising 1,4,7-triazacyclononanetriacetic acid chelate moiety (NOTA) were labeled with 67Ga or 68Ga. The autoradiography was conducted by incubating the tissue samples with the bispecific antibody TF2, followed by washing and incubation with one of the radiolabeled hapten molecules. After washing, drying and exposure to phosphor imager plates, the autoradiograms were analyzed and compared to standard histochemistry (hematoxylin-eosin). Pronounced binding was found in the tissue from colorectal cancer using the bispecific antibody TF2 and either of the haptens [67/68Ga]Ga-IMP461 and [67/68Ga]Ga-IMP485. Distinct binding was also detected in the epithelium of most samples of neighboring tissue, taken at a minimum of 10 cm from the site of the tumor. It is concluded that pretargeting CEA with the bispecific antibody TF2 followed by the addition of 67/68Ga-labeled hapten is extremely sensitive for visualizing this marker for colorectal cancer. This methodology is therefore a very specific complement to other histochemical techniques in the diagnosis of biopsies or in samples taken from surgery. Use of the pretargeting technique in vivo may also be an advance in diagnosing patients with colorectal cancer, either using 67Ga and SPECT or 68Ga and PET.

  • 126.
    Hallberg, Mathias
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
    Sandström, Anja
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry.
    From the Anti-Nociceptive Substance P Metabolite Substance P (1-7) to Small Peptidomimetics2018In: Current protein and peptide science, ISSN 1389-2037, E-ISSN 1875-5550, Vol. 19, no 11, p. 1038-1048Article, review/survey (Refereed)
    Abstract [en]

    Substance P (SP) is associated with pain and inflammatory processes and is released from terminals of specific sensory nerves. This undecapeptide that mediates its effects through the neurokinin type 1 (NK1) receptor, is rapidly degraded in vivo to smaller fragments. The heptapeptide SP(1-7) with a hitherto unknown receptor, is a major bioactive fragment and displays often opposite actions to those induced by SP. Hence, SP(1-7) elicits anti-nociceptive and anti-hyperalgesic effects. These observations have attracted a substantial interest and in this mini-review the efforts to transform the heptapeptide SP(1-7) into more drug-like small-molecule SP(1-7) peptidomimetics as a potential new class of analgesics are summarized. Structure-activity relationship studies and subsequent amidation of the C-terminal and truncations from the N-terminal of the heptapeptide delivered the bioactive dipeptide amide Gln-Phe-NH2 showing a high affinity at the SP(1-7) binding site. Similarly, endomorphin-2, an endogenous opioid ligand containing a C-terminal carboxamide group, demonstrated a high affinity at the SP(1-7) binding site. Endomorphin-2 subjected to truncations yielded the potent dipeptide amide Phe-Phe-NH2. Structural optimization of the latter furnished more drug-like high affinity ligands and among those a constrained cis-3-phenylpyrrolidine derivative that after peripheral administration produced a significant anti-allodynic effect in a mouse SNI model of neuropathic pain. This SP(1-7) peptidomimetic was as effective as SP(1-7) in alleviating mechanical allodynia in mice. Although, additional structural modifications are needed to achieve compounds exhibiting high/fair bioavailability after oral administration, the examples presented herein demonstrate that the bioactive peptides SP(1-7) and endomorphin-2 can be converted into low molecular weight compounds that are able to mimic the in vivo actions of the heptapeptide SP(1-7).

  • 127.
    Hallberg, Mathias
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
    Sumners, Colin
    Univ Florida, Coll Med, Dept Physiol & Funct Genom, Gainesville, FL 32611 USA.;McKnight Brain Inst, Gainesville, FL 32611 USA..
    Steckelings, U. Muscha
    Univ Southern Denmark, Inst Mol Med, Dept Cardiovasc & Renal Res, POB 5230, Odense, Denmark..
    Hallberg, Anders
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Small-molecule AT2 receptor agonists2018In: Medicinal research reviews (Print), ISSN 0198-6325, E-ISSN 1098-1128, Vol. 38, no 2, p. 602-624Article, review/survey (Refereed)
    Abstract [en]

    The discovery of the first selective, small-molecule ATR receptor (AT2R) agonist compound 21 (C21) (8) that is now extensively studied in a large variety of in vitro and in vivo models is described. The sulfonylcarbamate derivative 8, encompassing a phenylthiofen scaffold is the drug-like agonist with the highest affinity for the AT2R reported to date (K-i = 0.4 nM). Structure-activity relationships (SAR), regarding different biaryl scaffolds and functional groups attached to these scaffolds and with a particular focus on the impact of various para substituents displacing the methylene imidazole group of 8, are discussed. Furthermore, the consequences of migration of the methylene imidazole group and presumed structural requirements for ligands that are aimed as AT2R agonists (e.g. 8) or AT2R antagonists (e.g. 9), respectively, are briefly addressed. A summary of the pharmacological actions of C21 (8) is also presented.

  • 128.
    Han, Hongya
    et al.
    Karolinska Univ, Huddinge Hosp, Ctr Hematol & Regenerat Med, Karolinska Inst,Dept Med, Stockholm, Sweden.;Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Liang, Xiuming
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Ekberg, Monica
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Kritikou, Joanna S.
    Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Stockholm, Sweden..
    Brunnstrom, Asa
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Pelcman, Benjamin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Matl, Maria
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Clin Immunol & Allergy Unit, Stockholm, Sweden..
    Miao, Xinyan
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Clin Pharmacol Grp, Stockholm, Sweden..
    Andersson, Margareta
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Yuan, Xiaotian
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Schain, Frida
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Parvin, Selina
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Melin, Eva
    Danderyd Hosp, Karolinska Inst, Dept Clin Sci, Stockholm, Sweden..
    Sjoberg, Jan
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Xu, Dawei
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Westerberg, Lisa S.
    Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Stockholm, Sweden..
    Bjorkholm, Magnus
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Claesson, Hans-Erik
    Karolinska Univ, Hosp Solna, Karolinska Inst, Dept Med,Div Hematol, Stockholm, Sweden..
    Human 15-lipoxygenase-1 is a regulator of dendritic-cell spreading and podosome formation2017In: The FASEB Journal, ISSN 0892-6638, E-ISSN 1530-6860, Vol. 31, no 2, p. 491-504Article in journal (Refereed)
    Abstract [en]

    Dendritic cells (DCs) involved in proinflammatory immune responses derive mainly from peripheral monocytes, and the cells subsequently mature and migrate into the inflammatory micromilieu. Here we report that suppressing of 15-lipoxygenase-1 led to a substantial reduction in DC spreading and podosome formation in vitro. The surface expression of CD83 was significantly lower in both sh-15-lipoxygenase-1 (15-LOX-1)-transduced cells and DCs cultivated in the presence of a novel specific 15-LOX-1 inhibitor. The T-cell response against tetanus-pulsed DCs was only affected to a minor extent on inhibition of 15-LOX-1. In contrast, endocytosis and migration ability of DCs were significantly suppressed on 15-LOX-1 inhibition. The expression of 15-LOX-1 in DCs was also demonstrated in affected human skin in atopic and contact dermatitis, showing that the enzyme is indeed expressed in inflammatory diseases in vivo. This study demonstrated that inhibiting 15-LOX-1 led to an impaired podosome formation in DCs, and consequently suppressed antigen uptake and migration capacity. These results indicated that 15-LOX-1 is a potential target for inhibiting the trafficking of DCs to lymphoid organs and inflamed tissues and decreasing the inflammatory response attenuating symptoms of certain immunologic and inflammatory disorders such as dermatitis.-Han, H., Liang, X., Ekberg, M., Kritikou, J. S., Brunnstro " m, angstrom., Pelcman, B., Matl, M., Miao, X., Andersson, M., Yuan, X., Schain, F., Parvin, S., Melin, E., Sjoberg, J., Xu, D., Westerberg, L. S., Bjorkholm, M., Claesson, H.- E. Human 15-lipoxygenase- 1 is a regulator of dendritic-cell spreading and podosome formation.

  • 129.
    Hansson, Annelie
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Science.
    Structural Determination of Drug Metabolites from Doping Classed Compounds Using Mass Spectrometry2018Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Doping control in equine sports is important for a fair competition, but also to ensure the integrity of the betting system, as well as for animal welfare reasons. To detect the use of illicit compounds, screening for the parent compound is common. However, by using a metabolite as the analytical target instead, the detection time can be prolonged. For some compounds, the use of a metabolite is a necessity since the parent drug may not be detected at all.

    The metabolites of the selective androgen receptor modulators (SARM) S1, S4 and S22 were investigated in horse urine and plasma. The unchanged parent compounds had the longest detection time in plasma, but were not detected at all in urine. Instead, the longest detection time was measured for the metabolites 2-amino-5-nitro-4-(trifluoromethyl)phenyl hydrogen sulfate (SARMs S1 and S4) and 2-amino-5-cyano-4-(trifluoromethyl)phenyl hydrogen sulfate (SARM S22). These metabolites were thus suggested as analytical targets for doping control in urine while the parent compounds were suggested for plasma samples. 2-amino-5-nitro-4-(trifluoromethyl)phenyl hydrogen sulfate could also be produced in large quantities by the fungus Cunninghamella elegans to potentially be used as reference compound.

    The horse metabolites of the SARM LGD-4033 were also studied in urine and plasma. The formate adduct of LGD-4033 had the longest detection time in plasma and in urine after hydrolysis with β-glucuronidase. In non-hydrolyzed urine, the glucuronidated LGD-4033 was detected instead.

    Different in vitro models were used to predict in vivo metabolites of roxadustat, a hypoxia-inducible factor stabilizer. Cunninghamella elegans was successful in producing more metabolites compared to human and equine liver microsomes and human hepatocytes.

    The metabolite detection and identification in all experiments were accomplished using a UHPLC-Q-TOF MS instrument, where the high-resolution MS data was vital in determining which metabolites were formed.

    The thesis shows the benefits of investigating the metabolites of doping substances to allow for a successful doping control method in horse urine and plasma by prolonging the detection time. It also highlights the usefulness of Cunninghamella elegans as an alternative to the more commonly used in vitro models for both predicting and producing metabolites.

    List of papers
    1. Characterization of equine urinary metabolites of selective androgen receptor modulators (SARMs) S1, S4 and S22 for doping control purposes
    Open this publication in new window or tab >>Characterization of equine urinary metabolites of selective androgen receptor modulators (SARMs) S1, S4 and S22 for doping control purposes
    Show others...
    2015 (English)In: Drug Testing and Analysis, ISSN 1942-7603, E-ISSN 1942-7611, Vol. 7, no 8, p. 673-683Article in journal (Refereed) Published
    Abstract [en]

    Selective androgen receptor modulators, SARMs, constitute a class of compounds with anabolic properties but with few androgenic side-effects. This makes them possible substances of abuse and the World Anti-Doping Agency (WADA) has banned the entire class of substances. There have been several cases of illicit use of aryl propionamide SARMs in human sports and in 2013, 13 cases were reported. These substances have been found to be extensively metabolized in humans, making detection of metabolites necessary for doping control. SARMs are also of great interest to equine doping control, but the in vivo metabolite pattern and thus possible analytical targets have not been previously studied in this species. In this study, the urinary metabolites of the SARMs S1, S4, and S22 in horses were studied after intravenous injection, using ultra high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QToF-MS). Eight different metabolites were found for SARM S1, nine for SARM S4, and seven for SARM S22. The equine urinary metabolite profiles differed significantly from those of humans. The parent compounds were only detected for SARMs S4 and S22 and only at the first sampling time point at 3h post administration, making them unsuitable as target compounds. For all three SARMs tested, the metabolite yielding the highest response had undergone amide hydrolysis, hydroxylation and sulfonation. The resulting phase II metabolites (4-nitro-3-trifluoro-methyl-phenylamine sulfate for SARMs S1 and S4 and 4-cyano-3-trifluoro-methyl-phenylamine sulfate for SARM S22) are proposed as analytical targets for use in equine doping control.

    Keywords
    selective androgen receptor modulators, SARM, metabolite, equine, horse
    National Category
    Medicinal Chemistry
    Identifiers
    urn:nbn:se:uu:diva-261969 (URN)10.1002/dta.1768 (DOI)000359603700003 ()25560998 (PubMedID)
    Available from: 2015-09-08 Created: 2015-09-07 Last updated: 2018-03-14Bibliographically approved
    2. Investigation of the selective androgen receptor modulators S1, S4 and S22 and their metabolites in equine plasma using high-resolution mass spectrometry
    Open this publication in new window or tab >>Investigation of the selective androgen receptor modulators S1, S4 and S22 and their metabolites in equine plasma using high-resolution mass spectrometry
    Show others...
    2016 (English)In: Rapid Communications in Mass Spectrometry, ISSN 0951-4198, E-ISSN 1097-0231, Vol. 30, no 7, p. 833-842Article in journal (Refereed) Published
    Abstract [en]

    RationaleSelective androgen receptor modulators (SARMs) are prohibited in sports due to their performance enhancing ability. It is important to investigate the metabolism to determine appropriate targets for doping control. This is the first study where the equine metabolites of SARMs S1, S4 (Andarine) and S22 (Ostarine) have been studied in plasma. MethodsEach SARM was administered to three horses as an intravenous bolus dose and plasma samples were collected. The samples were pretreated with protein precipitation using cold acetonitrile before separation by liquid chromatography. The mass spectrometric analysis was performed using negative electrospray, quadrupole time-of-flight mass spectrometry operated in MSE mode and triple-quadrupole mass spectrometry operated in selected reaction monitoring mode. For the quantification of SARM S1, a deuterated analogue was used as internal standard. ResultsThe numbers of observed metabolites were eight, nine and four for the SARMs S1, S4 and S22, respectively. The major metabolite was formed by the same metabolic reactions for all three SARMs, namely amide hydrolysis, hydroxylation and sulfonation. The values of the determined maximum plasma concentrations were in the range of 97-170 ng/mL for SARM S1, 95-115 ng/mL for SARM S4 and 92-147 ng/mL for SARM S22 and the compounds could be detected for 96 h, 12 h and 18 h, respectively. ConclusionsThe maximum plasma concentration of SARMs S1, S4 and S22 was measured in the first sample (5 min) after administration and they were eliminated fast from plasma. The proposed targets to be used in equine doping control are the parent compounds for all three SARMs, but with the metabolite yielding the highest response as a complementary target. 

    National Category
    Biochemistry and Molecular Biology
    Identifiers
    urn:nbn:se:uu:diva-286622 (URN)10.1002/rcm.7512 (DOI)000372508100006 ()26969924 (PubMedID)
    Available from: 2016-04-28 Created: 2016-04-21 Last updated: 2018-03-14Bibliographically approved
    3. Equine in vivo-derived metabolites of the SARM LGD-4033 and comparison with human and fungal metabolites.
    Open this publication in new window or tab >>Equine in vivo-derived metabolites of the SARM LGD-4033 and comparison with human and fungal metabolites.
    Show others...
    2018 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 1074-1075, p. 91-98Article in journal (Refereed) Published
    Abstract [en]

    LGD-4033 has been found in human doping control samples and has the potential for illicit use in racehorses as well. It belongs to the pharmacological class of selective androgen receptor modulators (SARMs) and can stimulate muscle growth, much like anabolic steroids. However, SARMs have shown superior side effect profiles compared to anabolic steroids, which arguably makes them attractive for use by individuals seeking an unfair advantage over their competitors. The purpose of this study was to investigate the metabolites formed from LGD-4033 in the horse in order to find suitable analytical targets for doping controls. LGD-4033 was administered to three horses after which plasma and urine samples were collected and analyzed for metabolites using ultra high performance liquid chromatography coupled to a high resolution mass spectrometer. In horse urine, eight metabolites, both phase I and phase II, were observed most of which had not been described in other metabolic systems. Six of these were also detected in plasma. The parent compound was detected in plasma, but not in non-hydrolyzed urine. The longest detection times were observed for unchanged LGD-4033 in plasma and in urine hydrolyzed with β-glucuronidase and is thus suggested as the analytical target for doping control in the horse. The metabolite profile determined in the horse samples was also compared to those of human urine and fungal incubate from Cunninghamella elegans. The main human metabolite, dihydroxylated LGD-4033, was detected in the horse samples and was also produced by the fungus. However, it was a not a major metabolite for horse and fungus, which highlights the importance of performing metabolism studies in the species of interest.

    Keywords
    Doping, LGD-4033, Horse, Mass Spectrometry, Metabolite, SARM, Selective Androgen Receptor Modulator
    National Category
    Medicinal Chemistry
    Identifiers
    urn:nbn:se:uu:diva-344303 (URN)10.1016/j.jchromb.2017.12.010 (DOI)000425204900013 ()29334634 (PubMedID)
    Available from: 2018-03-06 Created: 2018-03-06 Last updated: 2018-05-07Bibliographically approved
    4. Investigation of the metabolites of the HIF stabilizer FG-4592 (roxadustat) in five different in vitro models and in a human doping control sample using high resolution mass spectrometry
    Open this publication in new window or tab >>Investigation of the metabolites of the HIF stabilizer FG-4592 (roxadustat) in five different in vitro models and in a human doping control sample using high resolution mass spectrometry
    Show others...
    2017 (English)In: Journal of Pharmaceutical and Biomedical Analysis, ISSN 0731-7085, E-ISSN 1873-264X, Vol. 134, p. 228-236Article in journal (Refereed) Published
    Abstract [en]

    FG-4592 is a hypoxia-inducible factor (HIF) stabilizer, which can increase the number of red blood cells in the body. It has not been approved by regulatory authorities, but is available for purchase on the Internet. Due to its ability to improve the oxygen transportation mechanism in the body, FG-4592 is of interest for doping control laboratories, but prior to this study, little information about its metabolism was available. In this study, the metabolism of FG-4592 was investigated in a human doping control sample and in five in vitro models: human hepatocytes and liver microsomes, equine liver microsomes and S9 fraction and the fungus Cunninghamella elegans. By using liquid chromatography coupled to a Q-TOF mass spectrometer operated in MSE and MSMS modes, twelve different metabolites were observed for FG-4592. One monohydroxylated metabolite was detected in both the human and equine liver microsome incubations. For the fungus Cunninghamella elegans eleven different metabolites were observed of which the identical monohydroxylated metabolite had the highest response. This rich metabolic profile and the higher levels of metabolites produced by Cunninghamella elegans demonstrates its usefulness as a metabolite producing medium. In the doping control urine sample, one metabolite, which was the result of a direct glucuronidation, was observed. No metabolites were detected in neither the human hepatocyte nor in the equine liver S9 fraction incubates.

    Keywords
    FG-4592, Drug metabolism, High resolution mass spectrometry
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-317588 (URN)10.1016/j.jpba.2016.11.041 (DOI)000392909900029 ()27918992 (PubMedID)
    Available from: 2017-03-24 Created: 2017-03-24 Last updated: 2018-03-14Bibliographically approved
  • 130.
    Hansson, Annelie
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Science.
    Knych, Heather
    Stanley, Scott
    Berndtson, Emma
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Science. Natl Vet Inst SVA, Dept Chem Environm & Feed Hyg, SE-75651 Uppsala, Sweden.
    Jackson, Liora
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Science. Natl Vet Inst SVA, Dept Chem Environm & Feed Hyg, SE-75651 Uppsala, Sweden.
    Bondesson, Ulf
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Science. Natl Vet Inst SVA, Dept Chem Environm & Feed Hyg, SE-75651 Uppsala, Sweden.
    Thevis, Mario
    Hedeland, Mikael
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Science. Natl Vet Inst SVA, Dept Chem Environm & Feed Hyg, SE-75651 Uppsala, Sweden.
    Equine in vivo-derived metabolites of the SARM LGD-4033 and comparison with human and fungal metabolites.2018In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 1074-1075, p. 91-98Article in journal (Refereed)
    Abstract [en]

    LGD-4033 has been found in human doping control samples and has the potential for illicit use in racehorses as well. It belongs to the pharmacological class of selective androgen receptor modulators (SARMs) and can stimulate muscle growth, much like anabolic steroids. However, SARMs have shown superior side effect profiles compared to anabolic steroids, which arguably makes them attractive for use by individuals seeking an unfair advantage over their competitors. The purpose of this study was to investigate the metabolites formed from LGD-4033 in the horse in order to find suitable analytical targets for doping controls. LGD-4033 was administered to three horses after which plasma and urine samples were collected and analyzed for metabolites using ultra high performance liquid chromatography coupled to a high resolution mass spectrometer. In horse urine, eight metabolites, both phase I and phase II, were observed most of which had not been described in other metabolic systems. Six of these were also detected in plasma. The parent compound was detected in plasma, but not in non-hydrolyzed urine. The longest detection times were observed for unchanged LGD-4033 in plasma and in urine hydrolyzed with β-glucuronidase and is thus suggested as the analytical target for doping control in the horse. The metabolite profile determined in the horse samples was also compared to those of human urine and fungal incubate from Cunninghamella elegans. The main human metabolite, dihydroxylated LGD-4033, was detected in the horse samples and was also produced by the fungus. However, it was a not a major metabolite for horse and fungus, which highlights the importance of performing metabolism studies in the species of interest.

  • 131.
    Hansson, Annelie
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Knych, Heather
    Stanley, Scott
    Thevis, Mario
    Bondesson, Ulf
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Hedeland, Mikael
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Characterization of equine urinary metabolites of selective androgen receptor modulators (SARMs) S1, S4 and S22 for doping control purposes2015In: Drug Testing and Analysis, ISSN 1942-7603, E-ISSN 1942-7611, Vol. 7, no 8, p. 673-683Article in journal (Refereed)
    Abstract [en]

    Selective androgen receptor modulators, SARMs, constitute a class of compounds with anabolic properties but with few androgenic side-effects. This makes them possible substances of abuse and the World Anti-Doping Agency (WADA) has banned the entire class of substances. There have been several cases of illicit use of aryl propionamide SARMs in human sports and in 2013, 13 cases were reported. These substances have been found to be extensively metabolized in humans, making detection of metabolites necessary for doping control. SARMs are also of great interest to equine doping control, but the in vivo metabolite pattern and thus possible analytical targets have not been previously studied in this species. In this study, the urinary metabolites of the SARMs S1, S4, and S22 in horses were studied after intravenous injection, using ultra high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QToF-MS). Eight different metabolites were found for SARM S1, nine for SARM S4, and seven for SARM S22. The equine urinary metabolite profiles differed significantly from those of humans. The parent compounds were only detected for SARMs S4 and S22 and only at the first sampling time point at 3h post administration, making them unsuitable as target compounds. For all three SARMs tested, the metabolite yielding the highest response had undergone amide hydrolysis, hydroxylation and sulfonation. The resulting phase II metabolites (4-nitro-3-trifluoro-methyl-phenylamine sulfate for SARMs S1 and S4 and 4-cyano-3-trifluoro-methyl-phenylamine sulfate for SARM S22) are proposed as analytical targets for use in equine doping control.

  • 132.
    Harang, Valérie
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Aspects of Optimisation of Separation of Drugs by Chemometrics2003Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Statistical experimental designs have been used for method development and optimisation of separation. Two reversed phase HPLC methods were optimised. Parameters such as the pH, the amount of tetrabutylammonium (TBA; co-ion) and the gradient slope (acetonitrile) were investigated and optimised for separation of erythromycin A and eight related compounds. In the second method, a statistical experimental design was used, where the amounts of acetonitrile and octane sulphonate (OSA; counter ion) and the buffer concentration were studied, and generation of an α-plot with chromatogram simulations optimised the separation of six analytes.

    The partial filling technique was used in capillary electrophoresis to introduce the chiral selector Cel7A. The effect of the pH, the ionic strength and the amount of acetonitrile on the separation and the peak shape of R- and S-propranolol were investigated.

    Microemulsion electrokinetic chromatography (MEEKC) is a technique similar to micellar electrokinetic chromatography (MEKC), except that the microemulsion has a core of tiny droplets of oil inside the micelles. A large number of factors can be varied when using this technique. A screening design using the amounts of sodium dodecyl sulphate (SDS), Brij 35, 1-butanol and 2-propanol, the buffer concentration and the temperature as factors revealed that the amounts of SDS and 2-propanol were the most important factors for migration time and selectivity manipulation of eight different compounds varying in charge and hydrophobicity. SDS and 2-propanol in the MEEKC method were further investigated in a three-level full factorial design analysing 29 different compounds sorted into five different groups. Different optimisation strategies were evaluated such as generating response surface plots of the selectivity/resolution of the most critical pair of peaks, employing chromatographic functions, simplex optimisation in MODDE and 3D resolution maps in DryLab™.

    Molecular descriptors were fitted in a PLS model to retention data from the three-level full factorial design of the MEEKC system. Two different test sets were used to study the predictive ability of the training set. It was concluded that 86 – 89% of the retention data could be predicted correctly for new molecules (80 – 120% of the experimental values) with different settings of SDS and 2-propanol.

    Statistical experimental designs and chemometrics are valuable tools for the development and optimisation of analytical methods. The same chemometric strategies can be employed for all types of separation techniques.

    List of papers
    1. Optimization of an HPLC Method for the Separation of Erythromycin and Related Compounds using Factorial Design
    Open this publication in new window or tab >>Optimization of an HPLC Method for the Separation of Erythromycin and Related Compounds using Factorial Design
    1999 In: Chromatographia, ISSN 0009-5893-99-11-525-07, Vol. 50, no 9/10, p. 525-531Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-90998 (URN)
    Available from: 2003-11-06 Created: 2003-11-06Bibliographically approved
    2. Liquid Chromatography Method Development and Optimization by Statistical Experimental Design and Chromatogram Simulations
    Open this publication in new window or tab >>Liquid Chromatography Method Development and Optimization by Statistical Experimental Design and Chromatogram Simulations
    2001 In: Chromatographia, ISSN 0009-5893-00-02-703-07, Vol. 54, no 11/12, p. 703-709Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-90999 (URN)
    Available from: 2003-11-06 Created: 2003-11-06Bibliographically approved
    3. A statistical experimental design to study factors affecting enantioseparation of propranolol by capillary electrophoresis with cellobiohydrolase (Cel7A) as chiral selector.
    Open this publication in new window or tab >>A statistical experimental design to study factors affecting enantioseparation of propranolol by capillary electrophoresis with cellobiohydrolase (Cel7A) as chiral selector.
    Show others...
    2002 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 23, no 14, p. 2306-2319Article in journal (Refereed) Published
    Abstract [en]

    The capillary electrophoretic enantioseparation of rac-propranolol using cellobiohydrolase Tr Cel7A as selector was optimized by an unbiased statistical experimental design. A set of pre-experiments was performed in order to identify critical experimental factors. In the definitive chemometric design pH, ranging from 5 to 7, ionic strength ranging between 0.01 and 0.02 and organic solvent additive in concentration from 1 to 19% v/v were studied. The response surface plot revealed a separation optimum in the pH interval studied. When all parameters were taken into account, a background electrolyte consisting of 0.016 M bistris-acetate buffer with pH 6.5 and 17% v/v acetonitrile gave the optimum separation. The significance of the statistical design was confirmed by the generally good agreement obtained between predicted response and actual experimental data.

    Keywords
    Capillary electrophoresis, chiral separation, CBHI, propranolol, statistical experimental design
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-91000 (URN)000177337000021 ()12210237 (PubMedID)
    Available from: 2003-11-06 Created: 2003-11-06 Last updated: 2017-12-14Bibliographically approved
    4. Microemulsion electrokinetic chromatography of drugs varying in charge and hydrophobicity: I. Impact of parameters on separation performance evaluated by multiple linear regression models
    Open this publication in new window or tab >>Microemulsion electrokinetic chromatography of drugs varying in charge and hydrophobicity: I. Impact of parameters on separation performance evaluated by multiple linear regression models
    Show others...
    2004 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 25, no 1, p. 80-93Article in journal (Refereed) Published
    Abstract [en]

    The separation of anionic, cationic and neutral drugs in microemulsion electrokinetic chromatography (MEEKC) was studied with a statistical experimental design. The concentration of sodium dodecyl sulfate (SDS, surfactant), 1-butanol (co-surfactant) and borate buffer and the factors Brij 35 (surfactant), 2-propanol (organic solvent) and cassette temperature were varied simultaneously, while the parameters pH (9.2), the concentration of octane (oil, 0.8% w/w), the voltage (10 kV) and the dimension of the fused-silica capillary, were kept constant. Eight different model substances were chosen with different hydrophobicities. Two of the analytes were positively charged, two were negatively charged, and the remaining four were neutral or close to neutral at the pH explored. The importance of each parameter on the separation window, the plate height and the retention factor for each of the analytes was studied by means of multiple linear regression (MLR) models. A new response was evaluated for anions, the quotient between the effective mobility in the microemulsion and the effective mobility in the corresponding buffer. Factors affecting selectivity changes were also explored, and it was found that SDS and 2-propanol had the largest effect on selectivity.

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-91001 (URN)10.1002/elps.200305671 (DOI)14730572 (PubMedID)
    Available from: 2003-11-06 Created: 2003-11-06 Last updated: 2017-12-14Bibliographically approved
    5. Microemulsion electrokinetic chromatography of drugs varying in charge and hydrophobicity: II. Strategies for optimisation of separation
    Open this publication in new window or tab >>Microemulsion electrokinetic chromatography of drugs varying in charge and hydrophobicity: II. Strategies for optimisation of separation
    2004 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 25, no 12, p. 1792-1809Article in journal (Refereed) Published
    Abstract [en]

    The separation of anionic, cationic, and neutral drugs in microemulsion electrokinetic chromatography (MEEKC) was studied. The concentration of sodium dodecyl sulfate (SDS; surfactant) and 2-propanol (organic solvent) was varied in a three-level full factorial design. 29 different model substances were chosen with different hydrophobicities and charges (neutral, positive, and negative). The models were calculated by means of multiple linear regression (MLR). The compounds were divided into five different subgroups, and different strategies for optimization of the separation within each group were investigated. The optimization was done by maximizing the selectivity using response surface plots in MODDE, by calculation of different chromatographic functions, and by using the software DryLab. For all the different groups, MODDE, almost all chromatographic functions and DryLab gave approximately the same settings of the factors for optimum separation. Attempts were made to fit descriptors of the compounds to the retention data from the three-level full factorial design by means of partial least squares projection to latent structures (PLS). Between 86 and 89% of all predictions of migration times were acceptable (80-120% of the observed value).

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-91002 (URN)10.1002/elps.200305812 (DOI)15213977 (PubMedID)
    Available from: 2003-11-06 Created: 2003-11-06 Last updated: 2017-12-14Bibliographically approved
  • 133.
    Hedeland, Mikael
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Pharmacy, Department of Pharmaceutical Chemistry.
    Cellobiohydrolase I as a chiral sector1999Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The protein cellobiohydrolase I (CBH I) immobilized on silica particles has been used as a chiral selector in liquid chromatography (LC) and the free enzyme has served as a chiral complexing agent in the background electrolyte in capillary electrophoresis (CE) using the partial tilling technique.

    The CE systems were much more efficient and more solutes could be enantioseparated than in the LC systems. Due to the high enantioselectivity, a few substances could be completely enantioresolved in an injected CBH I zone that was shorter than the sample zone. The affinity for the amino alcohol propranolol was very high at neutral pH and in order to displace this solute for detection, a plug of the disaccharide cellobiose was injected after the sample zone. The high affinity of propranolol combined with the displacing ability of cellobiose made it possible to preconcentrate the solute prior to enantioseparation by compression of long dilute sample zones.

    The mechanisms of retention and enantioselectivity in LC have been studied using charged and uncharged additives in the mobile phase. Site-specific mutation and covalent modification of the protein were also used to elucidate retention mechanisms. The enantioselective sites of amino alcohols and the acid warfarin overlap in the vicinity of the catalytically active site of CBH I. The main binding mechanism for both amines and acids was electrostatic attraction. Interestingly, both the retention and the enantioselectivity increased upon changing from potassium to sodium as mobile phase cation.

    The thermodynamics of the system was studied by microcalorimetry and LC. Both enantiomers of the amino alcohol alprenolol bind to CBH I with an endothermic enthalpy change. Hence, the complexation is entropy driven.

    Molecular mechanics calculations were carried out on the complex between propranolol and CBH I. The bound conformation of (S)-piopranolol differs both in geometry and energy compared to the preferred conformation in water solution. The ability of different parameter setups to reproduce the experimental structure of the (S)-propranolol - CBH I complex was investigated. One setup was chosen to suggest possible conformations of the (R)-enantiomer. The water structure in the bindning site seems to be an important factor in the complexation mechanism.

  • 134.
    Hedeland, Mikael
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Moura, Hercules
    Båverud, Viveca
    Woolfitt, Adrian R
    Bondesson, Ulf
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Barr, John R.
    Confirmation of botulism in birds and cattle by the mouse bioassay and Endopep-MS2011In: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 60, no 9, p. 1299-1305Article in journal (Refereed)
    Abstract [en]

    There have been several outbreaks of botulism among poultry and wild birds in Sweden in recent years. The National Veterinary Institute of Sweden (SVA) has identified botulinum neurotoxin (BoNT)/C1 or the mosaic BoNT/C1D using the mouse bioassay. This is believed to be the first report on the application of the Endopep mass spectrometry (Endopep-MS) method to selected clinical animal (serum and liver) samples and a feed sample that had previously given positive test results with the mouse bioassay. In the mouse bioassay eight of the eleven samples were found to be neutralized by both BoNT/C1 and /D antitoxins; the other three were neutralized only by BoNT/C1 antitoxin, but the mice showed a prolonged survival time when the samples had been treated with /D antitoxin. The Endopep-MS analysis, on the other hand, demonstrated only BoNT/C1 activity for all eleven samples. This suggests that at least eight of the samples were of the chimeric toxin type BoNT/C1D, where the enzymically active site is identical to that of BoNT/C1, while other parts of the protein contain sequences of BoNT/D. This is the first step of a cross-validation between the established mouse bioassay and the Endopep-MS of serotypes BoNT/C1 and /C1D. Endopep-MS is concluded to have potential as an attractive alternative to the mouse bioassay.

  • 135.
    Hedeland, Ylva
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Pettersson, Curt
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Chiral Separations in Nonaqueous Media2010In: Chiral Separations by Capillary Electrophoresis / [ed] Ann van Eeckhaut, Yvette Michotte, Boca Raton: CRC Press, 2010, 1.uppl., p. 271-312Chapter in book (Other academic)
  • 136.
    Hedeland, Ylva
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    Pettersson, Curt
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Pharmaceutical Chemistry.
    HPLC In Chiral Pharmaceutical Analysis2011In: Handbook of HPLC / [ed] Danilo Corradini, Boca Raton: CRC Press, 2011, 2nd ed., p. 507-534Chapter in book (Other academic)
  • 137.
    Holmberg, Pär
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Synthesis of Molecular Probes for Exploring the Human Consciousness, 5-HT7 Ligands and Salvinorins2005Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    In this study, we have addressed the serotonergic and the opioid system within the CNS. Both systems are of outmost importance in the etiology of disease states, especially mental disorders.

    In our investigation of the serotonergic system, we have synthesized novel enantiomerically pure 6-aryl-3-amino- and 8-aryl-3-aminochromans as ligands for the 5-HT7 receptor. One reason for the lack of understanding of the physiological functionality of the serotonin 5-HT7 receptor, the most recently discovered member of the serotonin receptor family, is the absence of partial agonists and agonists. In this series, we have identified partial agonists with more than189 fold selectivity over the 5-HT1A receptor and one agonist with 29 fold greater selectivity over the serotonin 5-HT1A receptor. Thus the present series constitutes a starting point for developing highly selective ligands for the 5-HT7 receptor.

    In our investigation of the opioid system, our focus has been on the natural product salvinorin A, which is a highly selective kappa opioid receptor agonist. In the total synthesis of salvinorin A, we have accomplished the synthesis of a key intermediate, 6-(3-furyl)-4-methyl-5,6-dihydro-pyran-2-one via ring closing metathesis. Furthermore, synthetic methodologies have been developed as a part of the total synthesis. Several lipases have been screeened for their ability to generate enantiomerically pure 1-(3-Furyl)-3-buten-1-ol via bio-catalyzed hydrolysis of the corresponding acetate. The lipase from Pseudomonas fluorescens was identified as having stereoselectivity high enough to generate a % ee value above 98%. We have also developed a route for the introduction of a hydroxyl functionality in the γ position of α,β-unsaturated cyclic ketones by the regioselective oxidation of 1-silyloxy-1,3-dienes using dimethyldioxirane. We have initiated the investigation of the pharmacophore responsible for the kappa opioid activity by synthesizing simplified analogues of salvinorin A. A synthetic route providing easy access to simplified analogues of salvinorin A have been established.

    List of papers
    1. Novel 2-Aminotetralin and 3-AminoChroman Derivatives as Selective Serotonin 5-HT7 Receptor Agonists and Antagonists
    Open this publication in new window or tab >>Novel 2-Aminotetralin and 3-AminoChroman Derivatives as Selective Serotonin 5-HT7 Receptor Agonists and Antagonists
    Show others...
    2004 In: Journal of Medicinal Chemistry, Vol. 47, p. 3927-3330Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-92717 (URN)
    Available from: 2005-03-10 Created: 2005-03-10Bibliographically approved
    2. Novel 3-aminochromans as potential pharmacological tools for the serotonin 5-HT7 receptor
    Open this publication in new window or tab >>Novel 3-aminochromans as potential pharmacological tools for the serotonin 5-HT7 receptor
    2005 In: Bioorganic & Medicinal Chemistry Letters, Vol. 15, no 747-750Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-92718 (URN)
    Available from: 2005-03-10 Created: 2005-03-10Bibliographically approved
    3. Enzymatic Kinetic Resolution of 1-(3-furyl)-3-buten-1ol
    Open this publication in new window or tab >>Enzymatic Kinetic Resolution of 1-(3-furyl)-3-buten-1ol
    2005 (English)In: Tetrahedron: asymmetry, ISSN 0957-4166, E-ISSN 1362-511X, Vol. 16, p. 2397-2399Article in journal (Other academic) Published
    Abstract [en]

    The enzymatic kinetic resolution of 1-(3-furyl)-3-buten-1-ol was investigated via the enantioselective hydrolysis of the corresponding acetate. Pseudomonas fluorescens (Fluka) was found to give the highest enantiomeric ratios of the 11 lipases screened. At 51% conversion, the ee value (eep) for the product was found to be 89%, giving an enantiomeric ratio (Ep) of 58, while the ee value (ees) for the substrate was 89%, giving an enantiomeric ratio (Ep) of 38.

    Place, publisher, year, edition, pages
    Elsevier, 2005
    National Category
    Organic Chemistry
    Research subject
    Chemistry with specialization in Organic Chemistry
    Identifiers
    urn:nbn:se:uu:diva-92719 (URN)10.1016/j.tetasy.2005.06.013 (DOI)
    Available from: 2005-03-10 Created: 2005-03-10 Last updated: 2017-12-14
    4. Regioselective formation and oxidation of 1-silyloxy-1,3-dienes from Hagemann’s ester and other conjugated enones, with dimethyldioxirane (DMDO) as oxidant.
    Open this publication in new window or tab >>Regioselective formation and oxidation of 1-silyloxy-1,3-dienes from Hagemann’s ester and other conjugated enones, with dimethyldioxirane (DMDO) as oxidant.
    Manuscript (Other academic)
    Identifiers
    urn:nbn:se:uu:diva-92720 (URN)
    Available from: 2005-03-10 Created: 2005-03-10 Last updated: 2010-01-13Bibliographically approved
  • 138.
    Holmberg, Pär
    et al.
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Pharmacy, Department of Medicinal Chemistry.
    Tedenborg, Lars
    Rosqvist, Susanne
    Johansson, Anette M
    Novel 3-aminochromans as potential pharmacological tools for the serotonin 5-HT7 receptor2005In: Bioorganic & Medicinal Chemistry Letters, ISSN 0960-894X, Vol. 15, no 3, p. 747-750Article in journal (Refereed)
  • 139.
    Holmboe, Michael
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Larsson, Per
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Anwar, Jamshed
    Bergström, Christel
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Colloidal Properties of Intestinal Fluid studied by Molecular Dynamics SimulationsManuscript (preprint) (Other academic)
  • 140.
    Holmboe, Michael
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy. Umea Univ, Dept Chem, Umea, Sweden..
    Larsson, Per
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Anwar, Jamshed
    Univ Lancaster, Chem Theory & Computat, Dept Chem, Lancaster LA1 4YB, England..
    Bergström, Christel A. S.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmacy.
    Partitioning into Colloidal Structures of Fasted State Intestinal Fluid Studied by Molecular Dynamics Simulations2016In: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 32, no 48, p. 12732-12740Article in journal (Refereed)
    Abstract [en]

    We performed molecular dynamics (MD) simulations to obtain insights into the structure and molecular interactions of colloidal structures present in fasted state intestinal fluid. Drug partitioning and interaction were studied with a mixed system of the bile salt taurocholate (TCH) and 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLiPC). Spontaneous aggregation of TCH and DLiPC from unconstrained MD simulations at the united-atom level using the Berger/Gromos54A7 force fields demonstrated that intermolecular hydrogen bonding between TCH molecules was an important factor in determining the overall TCH and DLiPC configuration. In bilayered systems, these intermolecular hydrogen bonds resulted in embedded transmembrane TCH clusters. Free energy simulations using the umbrella sampling technique revealed that the stability of these transmembrane TCH clusters was superior when they consisted of 3 or 4 TCH per bilayer leaflet. All-atom simulations using the Slipids/GAFF force fields showed that the TCH embedded in the bilayer decreased the energy barrier to penetrate the bilayer (Delta G(pen)) for water, ethanol, and carbamazepine, but not for the more lipophilic felodipine and danazol. This suggests that diffusion of hydrophilic to moderately lipophilic molecules through the bilayer is facilitated by the embedded TCH molecules. However, the effect of embedded TCH on the overall lipid/water partitioning was significant for danazol, indicating that the incorporation of TCH plays a crucial role for the partitioning of lipophilic solutes into e.g. lipidic vesicles existing in fasted state intestinal fluids. To conclude, the MD simulations revealed important intermolecular interactions in lipidic bilayers, both between the bile components themselves and with the drug molecules.

  • 141.
    Hovstadius, Peter
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Sciences, Clinical Pharmacology.
    Preclinical and Clinical Development of the Novel Cyanoguanidine CHS 828 for Cancer Treatment2005Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    CHS 828 is a cyanoguanidine with anti-tumour properties which has shown promising effects in several preclinical models. This thesis describes both preclinical and clinical studies aiming to investigate disease specific activity, clinical tolerability and efficacy of CHS 828.

    In paper I we investigated CHS 828 activity in a cell line panel with human myeloma cells, three of these cell-lines were also tested in vivo using a hollow fibre rat-model. In paper II we investigated CHS 828 activity in primary human tumour samples from patients. CHS 828 showed an effect on all tumour cell types tested both the primary human tumour samples and the myeloma cell lines. Notably, CHS 828 showed a high relative in vitro activity against tumour cells from chronic lymphocytic leukaemia and high-grade lymphoma.

    In a phase I trial we determined the maximum tolerated dose (MTD) of CHS 828. Haematological toxicity was generally mild and dominated by transient thrombocytopenia and lymphocytopenia. Non-haematological toxicity was mostly of gastrointestinal origin. The recommended phase two dose (RPTD) of CHS 828 was estimated to be 20 mg once daily for five days in cycles of 28 days duration.

    In a phase II trial we investigated the effect of CHS 828 on patients diagnosed with B-CLL. In total 12 patients were enrolled. CHS 828 was found to be well tolerated and the most common haematological toxicity was thrombocytopenia. Non-haematological toxicities were generally mild. Transient decreases in lymphocyte counts could be discerned coinciding with drug dosing, but no sustained clinical responses could be achieved.

    In conclusion, CHS 828 demonstrated marked effects in the preclinical investigations suggesting haematological malignancies as the main target. The clinical phase I study established a safe dose and the subsequent phase II trial in B-CLL patients showed biological effect but with no clinical disease response.

    List of papers
    1. Cytotoxic effect in vivo and in vitro of CHS 828 on human myeloma cell lines
    Open this publication in new window or tab >>Cytotoxic effect in vivo and in vitro of CHS 828 on human myeloma cell lines
    Show others...
    2004 In: Anti-Cancer Drugs, Vol. 15, no 1, p. 63-70Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-93817 (URN)
    Available from: 2005-11-24 Created: 2005-11-24Bibliographically approved
    2. Activity of CHS 828 in primary cultures of human hematological and solid tumors in vitro
    Open this publication in new window or tab >>Activity of CHS 828 in primary cultures of human hematological and solid tumors in vitro
    Show others...
    2001 In: Anti-Cancer Drugs, Vol. 12, no 10, p. 821-827Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-93818 (URN)
    Available from: 2005-11-24 Created: 2005-11-24Bibliographically approved
    3. A Phase I Study of CHS 828 in Patients with Solid Tumor Malignancy
    Open this publication in new window or tab >>A Phase I Study of CHS 828 in Patients with Solid Tumor Malignancy
    Show others...
    2002 (English)In: Clinical Cancer Research, ISSN 1078-0432, E-ISSN 1557-3265, Vol. 8, no 9, p. 2843-2850Article in journal (Refereed) Published
    Abstract [en]

    CHS 828 is a cyanoguanidine, which has demonstrated potent antitumor activity in preclinical tumor models. The activity of CHS 828 in vitro showed only low to moderate correlation to other antineoplastic agents suggesting a unique mechanism of action. Ten females and 6 males (median age 58 years) with solid tumors refractory to standard therapy were included in this Phase I study. The study drug was administered to fasting patients as a single oral dose on days 1–5 of each treatment cycle. Patients received one to six cycles of treatment. The doses ranged from 30 mg to 200 mg (total dose within a cycle). Hematological toxicity was generally mild and dominated by transient thrombocytopenia and lymphocytopenia. Nonhematological toxicity most frequently consisted of nausea, vomiting, diarrhea, fatigue, and localized genital mucositis. The dose-limiting toxicities were thrombocytopenia, thrombosis, esophagitis, diarrhea, and constipation. The recommended Phase II dose of CHS 828 was 20 mg once daily for 5 days in cycles of 28 days duration. The extent of systemic exposure of CHS 828 across patients was approximately dose proportional. The time at which the highest drug concentration occurs was 2.2 ± 1.3 h and half-life was 2.1 ± 0.52 h (mean ± SD). Large intra- and interindividual variation in dose level-adjusted maximum plasma concentration and the area under the curve from time 0 h to infinity were observed. There was an apparent inverse relationship between systemic exposure of CHS 828, and thrombocyte and lymphocyte nadir levels. No objective tumor responses were observed, and 7 patients showed stable disease after two courses of therapy.

    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-93819 (URN)12231525 (PubMedID)
    Available from: 2005-11-24 Created: 2005-11-24 Last updated: 2017-12-14Bibliographically approved
    4. A phase II study of the IkB kinase inhibitor CHS 828 in patients with chronic lymphocytic leukemia
    Open this publication in new window or tab >>A phase II study of the IkB kinase inhibitor CHS 828 in patients with chronic lymphocytic leukemia
    Show others...
    Article in journal (Refereed) Submitted
    Identifiers
    urn:nbn:se:uu:diva-93820 (URN)
    Available from: 2005-11-24 Created: 2005-11-24Bibliographically approved
  • 142.
    Hsu, Yu-Ming
    et al.
    Kaohsiung Med Univ, Coll Pharm, Grad Inst Nat Prod, Kaohsiung 80708, Taiwan..
    Chang, Fang-Rong
    Kaohsiung Med Univ, Coll Pharm, Grad Inst Nat Prod, Kaohsiung 80708, Taiwan.;Kaohsiung Med Univ, Ctr Infect Dis & Canc Res, Kaohsiung 80708, Taiwan.;Kaohsiung Med Univ, Res Ctr Environm Med, Kaohsiung 80708, Taiwan.;Natl Sun Yat Sen Univ, Dept Marine Biotechnol & Resources, Kaohsiung 80424, Taiwan.;Kaohsiung Med Univ Hosp, Ctr Canc, Kaohsiung 80708, Taiwan..
    Lo, I-Wen
    Kaohsiung Med Univ, Coll Pharm, Grad Inst Nat Prod, Kaohsiung 80708, Taiwan..
    Lai, Kuei-Hung
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy. Kaohsiung Med Univ, Coll Pharm, Grad Inst Nat Prod, Kaohsiung 80708, Taiwan..
    El-Shazly, Mohamed
    Ain Shams Univ, Fac Pharm, Dept Pharmacognosy & Nat Prod Chem, Cairo 11566, Egypt..
    Wu, Tung-Ying
    China Med Univ Hosp, Chinese Med Res & Dev Ctr, Taichung 40402, Taiwan. China Med Univ Hosp, Ctr Mol Med, Taichung 40402, Taiwan..
    Du, Ying-Chi
    Kaohsiung Med Univ, Coll Pharm, Grad Inst Nat Prod, Kaohsiung 80708, Taiwan..
    Hwang, Tsong-Long
    Chang Gung Univ Sci & Technol, Res Ctr Chinese Herbal Med, Res Ctr Ind Human Ecol, Taoyuan 33302, Taiwan.;Chang Gung Mem Hosp, Dept Anesthesiol, Taoyuan 33302, Taiwan. China Med Univ, Sch Pharm, Coll Pharm, Taichung 40402, Taiwan. China Med Univ, Res Ctr Chinese Herbal Med, Taichung 40402, Taiwan..
    Cheng, Yuan-Bin
    Kaohsiung Med Univ, Coll Pharm, Grad Inst Nat Prod, Kaohsiung 80708, Taiwan.;Kaohsiung Med Univ, Ctr Infect Dis & Canc Res, Kaohsiung 80708, Taiwan.;Kaohsiung Med Univ, Res Ctr Nat Prod & Drug Dev, Kaohsiung 80708, Taiwan..
    Wu, Yang-Chang
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy. Kaohsiung Med Univ, Coll Pharm, Grad Inst Nat Prod, Kaohsiung 80708, Taiwan.;Ain Shams Univ, Fac Pharm, Dept Pharmacognosy & Nat Prod Chem, Cairo 11566, Egypt.;China Med Univ Hosp, Chinese Med Res & Dev Ctr, Taichung 40402, Taiwan. China Med Univ Hosp, Ctr Mol Med, Taichung 40402, Taiwan.;Chang Gung Univ, Grad Inst Nat Prod, Coll Med, Taoyuan 33302, Taiwan..
    Zoanthamine-Type Alkaloids from the Zoanthid Zoanthus kuroshio Collected in Taiwan and Their Effects on Inflammation2016In: Journal of natural products (Print), ISSN 0163-3864, E-ISSN 1520-6025, Vol. 79, no 10, p. 2674-2680Article in journal (Refereed)
    Abstract [en]

    Zoanthus kuroshio is a colorful zoanthid with a fluorescent pink oral disc and brown tentacles, which dominates certain parts of the Taiwanese and Japanese coasts. This sea anemone is a rich source of biologically active alkaloids. In the current investigation, two novel halogenated zoanthamines [5 alpha-iodozoanthenamine (1) and 11 beta-chloro-11-deoxykuroshine A (2)], along with four new zoanthamines [18-epi-kuroshine A (3), 7 alpha-hydroxykuroshine E (4), 5 alpha-methoxykuroshine E (5), and 18-epi-kuroshine E (6)], and six known compounds were isolated from Z. kuroshio. Compounds 1 and 2 are the first examples of halogenated zoanthamine-type alkaloids isolated from nature. Compounds 3 and 6 are the first zoanthamine stereoisomers with a cis-junction of the A/B rings. All isolated compounds were evaluated for their anti-inflammatory activities by measuring their effects on superoxide anion generation and elastase release by human neutrophils in response to fMLP.

  • 143.
    Hugerth, Andreas M.
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Pharmacy, Department of Pharmaceutical Chemistry.
    Effects of polyelectrolyte conformation, charge density and ion specificity in polyelectrolyte and polyelectrolyte-drug interaction2000Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The central theme is how the structural characteristics of sulphated polyelectrolytes and theirinteraction with counter- and coions determine the degree of polyelectrolyte self-association (gelformation), interaction with oppositely charged polyelectrolyte and polyelectrolyte-amphiphilic druginteraction in protolytic solvent medium. Such knowledge is essential for understanding and regulatingthe interaction between polyelectrolytes as well as between polyelectrolytes and a drug substance in apharmaceutical formulation and for its fate in the human organism.

    The gel formation of the sulphated polyelectrolyte ê-carrageenan was found to be dependent on theextent of inter-chain helical association and thus the nature of the polyelectrolyte counterions. This wasshown from the relationship between the gel-sol melting temperature of the hydrophobic microdomains(T0), the fraction of polymer in helical conformation at T0, and the storage modulus of the samples inthe gel-state. Interaction between the oppositely charged polyelectrolytes carrageenan and chitosanresulted in the formation of polyelectrolyte complexes with a charge ratio of unity. However, in casesof inter helical association of carrageenan chitosan acted as a bridging element producing complexeswith a charge ratio below unity. This mechanism may be used to control the charge ratio ofpolyelectrolyte complexes.

    The solvation characteristics of the polyelectrolyte counterions also affected the polyelectrolyte-amphiphilic drug interaction (amitriptyline). The binding isotherm was shifted to a higher concentrationof free amphiphile according to the counterion sequence Li+ < Na+ < K+ < Rb+ ≈Cs+. The change inGibbs free energy per monomer amphiphile originating from exchanging the counterions (Li+ for Cs+),was of the order of kT. This is of the same order of magnitude as that obtained by significantlychanging the hydrophobicity of the amphiphilic drug molecule. Increasing the polyelectrolyte chargedensity decreased the critical aggregation concentration, increased the degree of cooperativity andincreased the magnitude of ion specific effects. Furthermore, the dependence of the. critical aggregationconcentration on ionic strength was linear and the polymer flexibility affected the efficiency of thepolycounterion properties of the polyelectrolyte. The binding isotherms, micropolarity, microviscosityand surface tension indicated the amitriptyline-polyelectrolyte interaction to be qualitatively andquantitatively similar to that observed for "typical" cationic surfactants.

  • 144.
    Hughes, Diarmaid
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Microbiology.
    Karlén, Anders
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry.
    Discovery and preclinical development of new antibiotics2014In: Upsala Journal of Medical Sciences, ISSN 0300-9734, E-ISSN 2000-1967, Vol. 119, no 2, p. 162-169Article, review/survey (Refereed)
    Abstract [en]

    Antibiotics are the medical wonder of our age, but an increasing frequency of resistance among key pathogens is rendering them less effective. If this trend continues the consequences for cancer patients, organ transplant patients, and indeed the general community could be disastrous. The problem is complex, involving abuse and overuse of antibiotics (selecting for an increasing frequency of resistant bacteria), together with a lack of investment in discovery and development (resulting in an almost dry drug development pipeline). Remedial approaches to the problem should include taking measures to reduce the selective pressures for resistance development, and taking measures to incentivize renewed investment in antibiotic discovery and development. Bringing new antibiotics to the clinic is critical because this is currently the only realistic therapy that can ensure the level of infection control required for many medical procedures. Here we outline the complex process involved in taking a potential novel antibiotic from the initial discovery of a hit molecule, through lead and candidate drug development, up to its entry into phase I clinical trials. The stringent criteria that a successful drug must meet, balancing high efficacy in vivo against a broad spectrum of pathogens, with minimal liabilities against human targets, explain why even with sufficient investment this process is prone to a high failure rate. This emphasizes the need to create a well-funded antibiotic discovery and development pipeline that can sustain the continuous delivery of novel candidate drugs into clinical trials, to ensure the maintenance of the advanced medical procedures we currently take for granted.

  • 145.
    Hultén, Johan