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Belfrage, Anna KarinORCID iD iconorcid.org/0000-0002-6257-4064
Alternative names
Publications (10 of 18) Show all publications
Belfrage, A. K., Abdurakhmanov, E., Åkerblom, E., Brandt, P., Alogheli, H., Neyts, J., . . . Johansson, A. (2018). Pan-NS3 protease inhibitors of hepatitis C virus based on an R3-elongated pyrazinone scaffold. European Journal of Medicinal Chemistry, 148, 453-464
Open this publication in new window or tab >>Pan-NS3 protease inhibitors of hepatitis C virus based on an R3-elongated pyrazinone scaffold
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2018 (English)In: European Journal of Medicinal Chemistry, ISSN 0223-5234, E-ISSN 1768-3254, Vol. 148, p. 453-464Article in journal (Refereed) Published
Abstract [en]

Herein, we present the design and synthesis of 2(1H)-pyrazinone based HCV NS3 protease inhibitors and show that elongated R-3 urea substituents were associated with increased inhibitory potencies over several NS3 protein variants. The inhibitors are believed to rely on beta-sheet mimicking hydrogen bonds which are similar over different genotypes and current drug resistant variants and correspond to the beta-sheet interactions of the natural peptide substrate. Inhibitor 36, for example, with a urea substituent including a cyclic imide showed balanced nanomolar inhibitory potencies against genotype la, both wild-type (K-i=30 nM) and R155K (K-i=2 nM), and genotype 3a (K-i=5 nM).

Place, publisher, year, edition, pages
Elsevier, 2018
Keywords
Hepatitis C, NS3, Genotype 3, Resistance, Pyrazinone
National Category
Medicinal Chemistry
Research subject
Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-340862 (URN)10.1016/j.ejmech.2018.02.032 (DOI)000428824700036 ()
Funder
Swedish Research Council, D0571301
Available from: 2018-02-04 Created: 2018-02-04 Last updated: 2018-05-31Bibliographically approved
Belfrage, A. K., Abdurakhmanov, E., Åkerblom, E., Brandt, P., Oshalim, A., Gising, J., . . . Sandström, A. (2016). Discovery of pyrazinone based compounds that potently inhibit the drug resistant enzyme variant R155K of the hepatitis C virus NS3 protease. Bioorganic & Medicinal Chemistry, 24(12), 2603-2620
Open this publication in new window or tab >>Discovery of pyrazinone based compounds that potently inhibit the drug resistant enzyme variant R155K of the hepatitis C virus NS3 protease
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2016 (English)In: Bioorganic & Medicinal Chemistry, ISSN 0968-0896, E-ISSN 1464-3391, Vol. 24, no 12, p. 2603-2620Article in journal (Refereed) Published
Abstract [en]

Herein, we present the design and synthesis of 2(1H)-pyrazinone based HCV NS3 protease inhibitors with variations in the C-terminus. Biochemical evaluation was performed using genotype 1a, both the wildtype and the drug resistant enzyme variant, R155K. Surprisingly, compounds without an acidic sulfonamide retained good inhibition, challenging our previous molecular docking model. Moreover, selected compounds in this series showed nanomolar potency against R155K NS3 protease; which generally confer resistance to all HCV NS3 protease inhibitors approved or in clinical trials. These results further strengthen the potential of this novel substance class, being very different to the approved drugs and clinical candidates, in the development of inhibitors less sensitive to drug resistance.

Keywords
Hepatitis C virus; Drug resistance; Pyrazinone; NS3 protease inhibitors; R155K
National Category
Organic Chemistry
Research subject
Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-243315 (URN)10.1016/j.bmc.2016.03.066 (DOI)000376727800002 ()27160057 (PubMedID)
Funder
Swedish Research Council, D0571301
Available from: 2015-02-08 Created: 2015-02-08 Last updated: 2017-12-04Bibliographically approved
Belfrage, A. K. (2015). Design and Synthesis of Hepatitis C Virus NS3 Protease Inhibitors: Targeting Different Genotypes and Drug-Resistant Variants. (Doctoral dissertation). Uppsala: Acta Universitatis Upsaliensis
Open this publication in new window or tab >>Design and Synthesis of Hepatitis C Virus NS3 Protease Inhibitors: Targeting Different Genotypes and Drug-Resistant Variants
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Since the first approved hepatitis C virus (HCV) NS3 protease inhibitors in 2011, numerous direct acting antivirals (DAAs) have reached late stages of clinical trials. Today, several combination therapies, based on different DAAs, with or without the need of pegylated interferon-α injection, are available for chronic HCV infections. The chemical foundation of the approved and late-stage HCV NS3 protease inhibitors is markedly similar. This could partly explain the cross-resistance that have emerged under the pressure of NS3 protease inhibitors. The first-generation NS3 protease inhibitors were developed to efficiently inhibit genotype 1 of the virus and were less potent against other genotypes.

The main focus in this thesis was to design and synthesize a new class of 2(1H)-pyrazinone based HCV NS3 protease inhibitors, structurally dissimilar to the inhibitors evaluated in clinical trials or approved, potentially with a unique resistance profile and with a broad genotypic coverage. Successive modifications were performed around the pyrazinone core structure to clarify the structure-activity relationship; a P3 urea capping group was found valuable for inhibitory potency, as were elongated R6 residues possibly directed towards the S2 pocket. Dissimilar to previously developed inhibitors, the P1’ aryl acyl sulfonamide was not essential for inhibition as shown by equally good inhibitory potency for P1’ truncated inhibitors. In vitro pharmacokinetic (PK) evaluations disclosed a marked influence from the R6 moiety on the overall drug-properties and biochemical evaluation of the inhibitors against drug resistant enzyme variants showed retained inhibitory potency as compared to the wild-type enzyme. Initial evaluation against genotype 3a displayed micro-molar potencies. Lead optimization, with respect to improved PK properties, were also performed on an advanced class of HCV NS3 protease inhibitors, containing a P2 quinazoline substituent in combination with a macro-cyclic proline urea scaffold with nano-molar cell based activities.

Moreover, an efficient Pd-catalyzed C-N urea arylation protocol, enabling high yielding introductions of advanced urea substituents to the C3 position of the pyrazinone, and a Pd-catalyzed carbonylation procedure, to obtain acyl sulfinamides, were developed. These methods can be generally applicable in the synthesis of bioactive compounds containing peptidomimetic scaffolds and carboxylic acid bioisosteres.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2015. p. 108
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Pharmacy, ISSN 1651-6192 ; 197
Keywords
hepatitis C virus, HCV, NS3 protease inhibitors, structure-activity relationship, 2(1H)-pyrazinone, quinazoline, resistance, Pd catalysis
National Category
Organic Chemistry Other Chemistry Topics
Research subject
Medicinal Chemistry; Pharmaceutical Science
Identifiers
urn:nbn:se:uu:diva-243317 (URN)978-91-554-9166-6 (ISBN)
Public defence
2015-03-27, B41 BMC, Husargatan 3, Uppsala, 09:15 (Swedish)
Opponent
Supervisors
Available from: 2015-03-05 Created: 2015-02-08 Last updated: 2015-03-12Bibliographically approved
Belfrage, A. K., Gising, J., Svensson, F., Åkerblom, E., Sköld, C. & Sandström, A. (2015). Efficient and Selective Palladium-Catalysed C-3 Urea Couplings to 3,5-Dichloro-2(1H)-pyrazinones. European Journal of Organic Chemistry (5), 978-986
Open this publication in new window or tab >>Efficient and Selective Palladium-Catalysed C-3 Urea Couplings to 3,5-Dichloro-2(1H)-pyrazinones
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2015 (English)In: European Journal of Organic Chemistry, ISSN 1434-193X, E-ISSN 1099-0690, no 5, p. 978-986Article in journal (Refereed) Published
Abstract [en]

The development of a robust palladium-catalysed urea N-arylation protocol to install various ureas at the 3-position of the 2(1H)-pyrazinone scaffold is described. The method involves Pd(OAc)2 in combination with bidentate ligands, xantphos [4,5-bis(diphenylphosphino)-9,9-dimethylxanthene] in particular, and resulted in good to excellent coupling yields of aliphatic, aromatic, and sterically hindered ureas. Furthermore, the C-3 chlorine was shown to be selectively displaced in the presence of aryl halide ureas, and this finding was supported by density functional theory (DFT) calculations. This allows further diversification of the scaffold for the production of compound libraries. Overall, the protocol facilitates further exploitation of pyrazinones as beta-sheet-inducing scaffolds in the development of sophisticated peptidomimetics/protease inhibitors. This is exemplified here by the synthesis of a new pyrazinone-based hepatitis C virus (HCV) NS3 protease inhibitor.

National Category
Organic Chemistry
Research subject
Chemistry with specialization in Organic Chemistry; Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-243254 (URN)10.1002/ejoc.201403405 (DOI)000349391700009 ()
Available from: 2015-02-06 Created: 2015-02-06 Last updated: 2017-12-04Bibliographically approved
Belfrage, A. K., Wakchaure, P., Larhed, M. & Sandström, A. (2015). Palladium-Catalyzed Carbonylation of Aryl Iodides with Sulfinamides. European Journal of Organic Chemistry (32), 7069-7074
Open this publication in new window or tab >>Palladium-Catalyzed Carbonylation of Aryl Iodides with Sulfinamides
2015 (English)In: European Journal of Organic Chemistry, ISSN 1434-193X, E-ISSN 1099-0690, no 32, p. 7069-7074Article in journal (Refereed) Published
Abstract [en]

A facile palladium(0)-catalyzed carbonylative protocol for the generation of new acyl-sulfinamides in moderate to good yields is described. Aliphatic and aromatic sulfinamides were exploited as hitherto unexplored nucleophiles in carbonylation chemistry, with use of CO gas generated ex situ from Mo(CO)6 in a sealed two-chamber system. Both electron-poor and electron-rich (hetero)aryl iodides were employed as electrophiles. The two-chamber system and the use of an inorganic base were essential for efficacious synthesis of acyl-sulfinamide products. Finally, it was demonstrated that a one-pot (or single-vial) synthesis of acyl-sulfinamides was feasible under CO at balloon pressure in the presence of Cs2CO3 as base.

Keywords
Homogenous catalysis; Carbonylation; Palladium; Molybdenum; Sulfinamides
National Category
Organic Chemistry
Identifiers
urn:nbn:se:uu:diva-265231 (URN)10.1002/ejoc.201500875 (DOI)000364532000012 ()
Available from: 2015-10-26 Created: 2015-10-26 Last updated: 2017-12-01Bibliographically approved
Gising, J., Belfrage, A. K., Alogheli, H., Ehrenberg, A., Åkerblom, E., Svensson, R., . . . Sandström, A. (2014). Achiral Pyrazinone-Based Inhibitors of the Hepatitis C Virus NS3 Protease and Drug-Resistant Variants with Elongated Substituents Directed Toward the S2 Pocket. Journal of Medicinal Chemistry, 57(5), 1790-1801
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
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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
Alogheli, H., Belfrage, A. K., Lampa, A., Gising, J., Åkerblom, E., Danielson, H., . . . Karlen, A. (2012). Binding mode prediction of HCV NS3A protease inhibitors. Abstract of Papers of the American Chemical Society, 244
Open this publication in new window or tab >>Binding mode prediction of HCV NS3A protease inhibitors
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2012 (English)In: Abstract of Papers of the American Chemical Society, ISSN 0065-7727, Vol. 244Article in journal (Refereed) Published
National Category
Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-212559 (URN)000324621806075 ()
Note

220XX 347-MEDI Times Cited:0 Cited References Count:0

Available from: 2013-12-11 Created: 2013-12-11 Last updated: 2017-12-06
Nilsson, M., Kalayanov, G., Winqvist, A., Pinho, P., Sund, C., Zhou, X.- . X., . . . Johansson, N. G. (2012). Discovery of 4 '-azido-2 '-deoxy-2 '-C-methyl cytidine and prodrugs thereof: A potent inhibitor of Hepatitis C virus replication. Bioorganic & Medicinal Chemistry Letters, 22(9), 3265-3268
Open this publication in new window or tab >>Discovery of 4 '-azido-2 '-deoxy-2 '-C-methyl cytidine and prodrugs thereof: A potent inhibitor of Hepatitis C virus replication
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2012 (English)In: Bioorganic & Medicinal Chemistry Letters, ISSN 0960-894X, E-ISSN 1090-2120, Vol. 22, no 9, p. 3265-3268Article in journal (Refereed) Published
Abstract [en]

4'-Azido-2'-deoxy-2'-methylcytidine (14) is a potent nucleoside inhibitor of the HCV NS5B RNA-dependent RNA polymerase, displaying an EC50 value of 1.2 mu M and showing moderate in vivo bioavailability in rat (F = 14%). Here we describe the synthesis and biological evaluation of 4'-azido-2'-deoxy-2'-methylcytidine and prodrug derivatives thereof. (C) 2012 Elsevier Ltd. All rights reserved.

Keywords
hepatitis c virus, ns5b polymerase, antivirals, nucleoside, prodrug, phosphoramidate, genotype 1 infection, antiviral activity, viral-hepatitis, analogs, design, 4'-azidocytidine, boceprevir, telaprevir, resistance
National Category
Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-212500 (URN)10.1016/j.bmcl.2012.03.021 (DOI)000302964300045 ()
Note

928EN Times Cited:1 Cited References Count:21

Available from: 2013-12-11 Created: 2013-12-11 Last updated: 2017-12-06Bibliographically approved
Simmen, K. A., De Kock, H. A., Raboisson,, P. J., Hu, L., Tahri, A., Surleraux,, D. L., . . . Vendeville,, S. M. (2012). Macrocylic inhibitors of hepatitis C virus. US Patent 8349869.
Open this publication in new window or tab >>Macrocylic inhibitors of hepatitis C virus
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2012 (English)Patent (Other (popular science, discussion, etc.))
Abstract [en]

Inhibitors of HCV replication of formula (I)

and the N-oxides, salts, and stereoisomers, wherein

  • each dashed line represents an optional double bond;
  • X is N, CH and where X bears a double bond it is C;
  • R1 is —OR7, —NH—SO2R8;
  • R2 is hydrogen, and where X is C or CH, R2 may also be C1-6alkyl;
  • R3 is hydrogen, C1-6alkyl, C1-6alkoxyC1-6alkyl, C3-7cycloalkyl;
  • R4 is aryl or Het; n is 3, 4, 5, or 6;
  • R5 is halo, C1-6alkyl, hydroxy, C1-6alkoxy, phenyl, or Het;
  • R6 is C1-6alkoxy, or dimethylamino;
  • R7 is hydrogen; aryl; Het; C3-7cycloalkyl optionally substituted with C1-6alkyl; or C1-6alkyl optionally substituted with C3-7cycloalkyl, aryl or with Het;
  • R8 is aryl; Het; C3-7cycloalkyl optionally substituted with C1-6alkyl; or C1-6alkyl optionally substituted with C3-7cycloalkyl, aryl or with Het;
  • aryl is phenyl optionally substituted with one, two or three substituents;
  • Het is a 5 or 6 membered saturated, partially unsaturated or completely unsaturated heterocyclic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, and being optionally substituted with one, two or three substituents;
  • pharmaceutical compositions containing compounds (I) and processes for preparing compounds (I). Bioavailable combinations of the inhibitors of HCV of formula (I) with ritonavir are also provided.
National Category
Medicinal Chemistry
Research subject
Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-212549 (URN)
Patent
US Patent 8349869
Available from: 2013-12-11 Created: 2013-12-11 Last updated: 2018-01-11
Ayesa, S., Belfrage, A. K., Classon, B., Grabowska, U., Hewitt, E., Ivanov, V., . . . Wähling, H. (2011). CYSTEINE PROTEASE INHIBITORS. US Patent App.20110312964.
Open this publication in new window or tab >>CYSTEINE PROTEASE INHIBITORS
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2011 (English)Patent (Other (popular science, discussion, etc.))
Abstract [en]

Compounds of the formula I

wherein

R1a is H; and R1b is C1-C6 alkyl, Carbocyclyl or Het; or

R1a and R1b together define a saturated cyclic amine with 3-6 ring atoms;

R2a and R2b are H, halo, C1-C4alkyl, C1-C4haloalkyl, C1-C4alkoxy; or

R2a and R2b together with the carbon atom to which they are attached form a C3-C6cycloalkyl;

R3 is a branched C5-C10alkyl chain, C2-C4haloalkyl or C3-C7cycloalkylmethyl,

R4 is Het, Carbocyclyl,

optionally substituted as defined in the specification and pharmaceutically acceptable salts,

hydrates and N-oxides thereof; are inhibitors of cathepsin S and have utility in the treatment of psoriasis, autoimmune disorders and other disorders such as asthma, arteriosclerosis, COPD and chronic pain.

National Category
Medicinal Chemistry
Research subject
Medicinal Chemistry
Identifiers
urn:nbn:se:uu:diva-212554 (URN)
Patent
US Patent App.20110312964
Available from: 2013-12-11 Created: 2013-12-11 Last updated: 2018-01-11
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-6257-4064

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