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Lindeberg, Gunnar
Publications (10 of 33) Show all publications
Skogh, A., Lesniak, A., Gaugaz, F. Z., Svensson, R., Lindeberg, G., Fransson, R., . . . Sandström, A. (2017). Impact of N-methylation of the substance P 1-7 amide on anti-allodynic effect in mice after peripheral administration. European Journal of Pharmaceutical Sciences, 109, 533-540, Article ID S0928-0987(17)30497-9.
Open this publication in new window or tab >>Impact of N-methylation of the substance P 1-7 amide on anti-allodynic effect in mice after peripheral administration
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2017 (English)In: European Journal of Pharmaceutical Sciences, ISSN 0928-0987, E-ISSN 1879-0720, Vol. 109, p. 533-540, article id S0928-0987(17)30497-9Article in journal (Refereed) Published
Abstract [en]

Substance P 1-7 (SP1-7, Arg1-Pro2-Lys3-Pro4-Gln5-Gln6-Phe7) is the major bioactive metabolite formed after proteolytic degradation of the tachykinin substance P (SP). This heptapeptide often opposes the effects of the mother peptide. Hence, SP1-7 is having anti-inflammatory, anti-nociceptive and anti-hyperalgesic effects in experimental models. Despite all encouraging properties of SP1-7 its exact mode of action has not yet been elucidated which has hampered further development of this heptapeptide in drug discovery. Contrary to SP that mediates its biological activity via the NK-1 receptor, the N-terminal fragment SP1-7 acts through an unknown target that is distinct from all known opioid and tachykinin receptors. The SP1-7 amide 1 (Arg1-Pro2-Lys3-Pro4-Gln5-Gln6-Phe7-NH2) was previously shown to be superior to the endogenous SP1-7 in all experimental pain models where the two compounds were compared. Herein, we report that N-methylation scan of the backbone of the SP1-7 amide (1) results in peptides that are significantly less prone to undergo proteolysis in plasma from both mouse and human. However, with the two exceptions of the [MeLys3]SP1-7 amide (3) and the [MeGln5]SP1-7 amide (4), the peptides with a methyl group attached to the backbone are devoid of significant anti-allodynic effects after peripheral administration in the spared nerve injury (SNI) mouse model of neuropathic pain. It is suggested that the N-methylation does not allow these peptides to form the accurate bioactive conformations or interactions required for efficient binding to the macromolecular target. The importance of intact N-terminal Arg1 and C-terminal Phe7, anticipated to serve as address and message residues, respectively, for achieving the anti-allodynic effect is emphasized. Notably, the three heptapeptides: the SP1-7 amide (1), the [MeLys3]SP1-7 amide (3) amide and the [MeGln5]SP1-7 amide (4) are all considerably more effective in the SNI mouse model than gabapentin that is widely used in the clinic for treatment of neuropathic pain.

Keywords
Anti-allodynia, N-methylation, SP(1–7) amide, Solid phase peptide synthesis (SPPS), Spared nerve injury (SNI), Substance P (SP)
National Category
Organic Chemistry
Identifiers
urn:nbn:se:uu:diva-335651 (URN)10.1016/j.ejps.2017.09.007 (DOI)000413325000055 ()28887235 (PubMedID)
Funder
Swedish Research Council, 9459The Swedish Brain FoundationBerzelii Centre EXSELENT
Available from: 2017-12-07 Created: 2017-12-07 Last updated: 2018-02-28
Skogh, A., Lesniak, A., Gaugaz, F. Z., Svensson, R., Lindeberg, G., Fransson, R., . . . Sandström, A. (2017). Importance of N-and C-terminal residues of substance P 1-7 for alleviating allodynia in mice after peripheral administration. European Journal of Pharmaceutical Sciences, 106, 345-351
Open this publication in new window or tab >>Importance of N-and C-terminal residues of substance P 1-7 for alleviating allodynia in mice after peripheral administration
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2017 (English)In: European Journal of Pharmaceutical Sciences, ISSN 0928-0987, E-ISSN 1879-0720, Vol. 106, p. 345-351Article in journal (Refereed) Published
Abstract [en]

The heptapeptide SP1-7 (1, Arg(1)-Pro(2)-Lys(3)-Pro(4)-Gln(5)-Gln(6)-Phe(7)) is the major bioactive metabolite formed after proteolytic processing of the neuropeptide substance P (SP, Arg(1)-Pro(2)-Lys(3)-Pro(4)-GIn(5)-Gln(6)-Phe(7)-Phe(8)-Gly(9)-Leu(10)-Meti(11)-NH2). The heptapeptide 1 frequently exhibits opposite effects to those induced by SP, such as exerting antinociception, or attenuating thermal hyperalgesia and mechanical allodynia. The heptapeptide SP1-7 amide (2, Arg(1)-Pro(2)-Lys(3)-Pro(4)-Gln(5)-Gln(6)-Phe(7)-NH2 ) is often more efficacious than 1 in experimental pain models. We have now assessed the anti-allodynic outcome after systemic administration of 2 and a series of Ala substituted and truncated analogues of 2, in the spared nerve injury (SNI) mice model and the results obtained were correlated with in vitro plasma stability and permeability measurements. It is herein demonstrated that an intact Arg(1) in SP1-7 amide analogues is fundamental for retaining a potent in vivo effect, while Lys(3) of 2 is less important. A displacement with Ala(1) or truncation rendered the peptide analogues either inactive or with a significantly attenuated in vivo activity. Thus, the pentapeptide SP3-7 amide (7, t(1/2) = 11.1 min) proven to be the major metabolite of 2, demonstrated an in vivo effect itself although considerably less significant than 2 in the SNI model. Intraperitoneal administration of 2 in a low dose furnished the most powerful anti-allodynic effect in the SNI model of all the analogous evaluated, despite a fast proteolysis of 2 in plasma (t(1/2) = 6.4 min). It is concluded that not only the C-terminal residue, that we previously demonstrated, but also the N-terminal with its basic side chain, are important for achieving effective pain relief. This information is of value for the further design process aimed at identifying more drug-like SP1-7 amide related peptidomimetics with pronounced antiallodynic effects.

Place, publisher, year, edition, pages
Elsevier, 2017
Keywords
Neuropathic pain, Spared nerve injury (SNI), SP1-7, Neuropeptides, Plasma stability, Structure-activity relationship, Message-address concept
National Category
Pharmacology and Toxicology
Identifiers
urn:nbn:se:uu:diva-334033 (URN)10.1016/j.ejps.2017.06.004 (DOI)000406988600036 ()28587787 (PubMedID)
Available from: 2017-11-23 Created: 2017-11-23 Last updated: 2018-02-28Bibliographically approved
Mitran, B., Varasteh, Z., Selvaraju, R. K., Lindeberg, G., Sörensen, J., Larhed, M., . . . Orlova, A. (2016). Selection of optimal chelator improves the contrast of GRPR imaging using bombesin analogue RM26.. International journal of oncology, 48(5), 2124-2134
Open this publication in new window or tab >>Selection of optimal chelator improves the contrast of GRPR imaging using bombesin analogue RM26.
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2016 (English)In: International journal of oncology, ISSN 1791-2423, Vol. 48, no 5, p. 2124-2134Article in journal (Refereed) Published
Abstract [en]

Bombesin (BN) analogs bind with high affinity to gastrin-releasing peptide receptors (GRPRs) that are up-regulated in prostate cancer and can be used for the visualization of prostate cancer. The aim of this study was to investigate the influence of radionuclide-chelator complexes on the biodistribution pattern of the 111In-labeled bombesin antagonist PEG2-D-Phe-Gln-Trp-Ala-Val-Gly-His-Sta-Leu-NH2 (PEG2-RM26) and to identify an optimal construct for SPECT imaging. A series of RM26 analogs N-terminally conjugated with NOTA, NODAGA, DOTA and DOTAGA via a PEG2 spacer were radiolabeled with 111In and evaluated both in vitro and in vivo. The conjugates were successfully labeled with 111In with 100% purity and retained binding specificity to GRPR and high stability. The cellular processing of all compounds was characterized by slow internalization. The IC50 values were in the low nanomolar range, with lower IC50 values for positively charged natIn-NOTA-PEG2-RM26 (2.6±0.1 nM) and higher values for negatively charged natIn-DOTAGA-PEG2-RM26 (4.8±0.5 nM). The kinetic binding studies showed KD values in the picomolar range that followed the same pattern as the IC50 data. The biodistribution of all compounds was studied in BALB/c nu/nu mice bearing PC-3 prostate cancer xenografts. Tumor targeting and biodistribution studies displayed rapid clearance of radioactivity from the blood and normal organs via kidney excretion. All conjugates showed similar uptake in tumors at 4 h p.i. The radioactivity accumulation in GRPR-expressing organs was significantly lower for DOTA- and DOTAGA-containing constructs compared to those containing NOTA and NODAGA. 111In-NOTA-PEG2-RM26 with a positively charged complex showed the highest initial uptake and the slowest clearance of radioactivity from the liver. At 4 h p.i., DOTA- and DOTAGA-coupled analogs showed significantly higher tumor-to-organ ratios compared to NOTA- and NODAGA-containing variants. The NODAGA conjugate demonstrated the best retention of radioactivity in tumors, and, at 24 h p.i., had the highest contrast to blood, muscle and bones.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-281358 (URN)10.3892/ijo.2016.3429 (DOI)000372568600037 ()26983776 (PubMedID)
Funder
Swedish Cancer SocietySwedish Research Council
Available from: 2016-03-23 Created: 2016-03-23 Last updated: 2016-04-28Bibliographically approved
Mitran, B., Varasteh, Z., Rosenström, U., Tolmachev, V., Lindeberg, G., Larhed, M. & Orlova, A. (2015). Development of radiocobalt-labeled GRPR antagonist NOTA-PEG2-RM26.. Paper presented at 28th Annual Congress of the European-Association-of-Nuclear-Medicine (EANM), OCT 10-14, 2015, Hamburg, GERMANY. European Journal of Nuclear Medicine and Molecular Imaging, 42(S1), S142-S142
Open this publication in new window or tab >>Development of radiocobalt-labeled GRPR antagonist NOTA-PEG2-RM26.
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2015 (English)In: European Journal of Nuclear Medicine and Molecular Imaging, ISSN 1619-7070, E-ISSN 1619-7089, Vol. 42, no S1, p. S142-S142Article in journal, Meeting abstract (Other academic) Published
National Category
Radiology, Nuclear Medicine and Medical Imaging
Identifiers
urn:nbn:se:uu:diva-269141 (URN)000363013201248 ()
Conference
28th Annual Congress of the European-Association-of-Nuclear-Medicine (EANM), OCT 10-14, 2015, Hamburg, GERMANY
Note

Meeting Abstract: OP340

Available from: 2015-12-18 Created: 2015-12-14 Last updated: 2017-12-01Bibliographically approved
Orlova, A., Varasteh, Z., Mitran, B., Rosestedt, M., Velikyan, I., Rosenström, U., . . . Tolmachev, V. (2015). Influence of chelators on targeting properties of In-111 and Ga-68 labeled GRPR antagonist. Journal of labelled compounds & radiopharmaceuticals, 58, S88-S88
Open this publication in new window or tab >>Influence of chelators on targeting properties of In-111 and Ga-68 labeled GRPR antagonist
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2015 (English)In: Journal of labelled compounds & radiopharmaceuticals, ISSN 0362-4803, E-ISSN 1099-1344, Vol. 58, p. S88-S88Article in journal, Meeting abstract (Other academic) Published
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-284894 (URN)000369950200089 ()
Available from: 2016-04-19 Created: 2016-04-19 Last updated: 2017-11-30Bibliographically approved
Varasteh, Z., Mitran, B., Rosenström, U., Velikyan, I., Rosestedt, M., Lindeberg, G., . . . Orlova, A. (2015). The effect of macrocyclic chelators on the targeting properties of the 68Ga-labeled gastrin releasing peptide receptor antagonist PEG2-RM26. Nuclear Medicine and Biology, 42(5), 446-454
Open this publication in new window or tab >>The effect of macrocyclic chelators on the targeting properties of the 68Ga-labeled gastrin releasing peptide receptor antagonist PEG2-RM26
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2015 (English)In: Nuclear Medicine and Biology, ISSN 0969-8051, E-ISSN 1872-9614, Vol. 42, no 5, p. 446-454Article in journal (Refereed) Published
Abstract [en]

Introduction

Overexpression of gastrin-releasing peptide receptors (GRPR) has been reported in several cancers. Bombesin (BN) analogs are short peptides with a high affinity for GRPR. Different BN analogs were evaluated for radionuclide imaging and therapy of GRPR-expressing tumors. We have previously investigated an antagonistic analog of BN (D-Phe-Gln-Trp-Ala-Val-Gly-His-Sta-Leu-NH2, RM26) conjugated to NOTA via a PEG2 spacer (NOTA-PEG2-RM26) labeled with 68Ga, 111In and Al18F. 68Ga-labeled NOTA-PEG2-RM26 showed high tumor-to-organ ratios.

Methods

The influence of different macrocyclic chelators (NOTA, NODAGA, DOTA and DOTAGA) on the targeting properties of 68Ga-labeled PEG2-RM26 was studied in vitro and in vivo.

Results

All conjugates were labeled with generator-produced 68Ga with high yields and demonstrated high stability and specific binding to GRPR. The IC50 values of natGa-X-PEG2-RM26 (X = NOTA, DOTA, NODAGA, DOTAGA) were 2.3 ± 0.2, 3.0 ± 0.3, 2.9 ± 0.3 and 10.0 ± 0.6 nM, respectively. The internalization of the conjugates by PC-3 cells was low. However, the DOTA-conjugated analog demonstrated a higher internalization rate compared to other analogs. GRPR-specific uptake was found in receptor-positive normal tissues and PC-3 xenografts for all conjugates. The biodistribution of the conjugates was influenced by the choice of the chelator moiety. Although all radiotracers cleared rapidly from the blood, [68Ga]Ga-NOTA-PEG2-RM26 showed significantly lower uptake in lung, muscle and bone compared to the other analogs. The uptake in tumors (5.40 ± 1.04 %ID/g at 2 h p.i.) and the tumor-to-organ ratios (25 ± 3, 157 ± 23 and 39 ± 4 for blood, muscle and bone, respectively) were significantly higher for the NOTA-conjugate than the other analogs.

Conclusions

Chelators had a clear influence on the biodistribution and targeting properties of 68Ga-labeled antagonistic BN analogs. Positively charged [68Ga]Ga-NOTA-PEG2-RM26 provided a low kidney radioactivity uptake, high affinity, high tumor uptake and high image contrast.

National Category
Radiology, Nuclear Medicine and Medical Imaging
Identifiers
urn:nbn:se:uu:diva-232120 (URN)10.1016/j.nucmedbio.2014.12.009 (DOI)000353369000005 ()25684649 (PubMedID)
Funder
Swedish Cancer SocietySwedish Research Council
Available from: 2014-09-12 Created: 2014-09-12 Last updated: 2017-12-05Bibliographically approved
Mitran, B., Varasteh, Z., Rosestedt, M., Velikyan, I., Rosenström, U., Lindeberg, G., . . . Orlova, A. (2014). Influence of chelators on biodistribution and targeting properties of GRPR antagonist. Paper presented at Annual Congress of the European-Association-of-Nuclear-Medicine (EANM), OCT 18-22, 2014, Gothenburg, SWEDEN. European Journal of Nuclear Medicine and Molecular Imaging, 41(S2), S320-S320, Article ID PW012.
Open this publication in new window or tab >>Influence of chelators on biodistribution and targeting properties of GRPR antagonist
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2014 (English)In: European Journal of Nuclear Medicine and Molecular Imaging, ISSN 1619-7070, E-ISSN 1619-7089, Vol. 41, no S2, p. S320-S320, article id PW012Article in journal, Meeting abstract (Other academic) Published
National Category
Radiology, Nuclear Medicine and Medical Imaging
Identifiers
urn:nbn:se:uu:diva-247697 (URN)000348841900527 ()
Conference
Annual Congress of the European-Association-of-Nuclear-Medicine (EANM), OCT 18-22, 2014, Gothenburg, SWEDEN
Available from: 2015-03-31 Created: 2015-03-23 Last updated: 2017-12-04Bibliographically approved
Lampa, A., Bergman, S., Svahn Gustafsson, S., Alogheli, H., Åkerblom, E., Lindeberg, G., . . . Sandström, A. (2014). Novel Peptidomimetic Hepatitis C Virus NS3/4A Protease Inhibitors Spanning the P2–P1′ Region [Letter to the editor]. ACS Medicinal Chemistry Letters, 5(3), 249-254
Open this publication in new window or tab >>Novel Peptidomimetic Hepatitis C Virus NS3/4A Protease Inhibitors Spanning the P2–P1′ Region
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2014 (English)In: ACS Medicinal Chemistry Letters, ISSN 1948-5875, E-ISSN 1948-5875, Vol. 5, no 3, p. 249-254Article in journal, Letter (Refereed) Published
Abstract [en]

Herein, novel hepatitis C virus NS3/4A protease inhibitors based on a P2 pyrimidinyloxyphenylglycine in combination with various regioisomers of an aryl acyl sulfonamide functionality in P1 are presented. The P1′ 4-(trifluoromethyl)phenyl side chain was shown to be particularly beneficial in terms of inhibitory potency. Several inhibitors with Ki-values in the nanomolar range were developed and included identification of promising P3-truncated inhibitors spanning from P2–P1′. Of several different P2 capping groups that were evaluated, a preference for the sterically congested Boc group was revealed. The inhibitors were found to retain inhibitory potencies for A156T, D168V, and R155K variants of the protease. Furthermore, in vitro pharmacokinetic profiling showed several beneficial effects on metabolic stability as well as on apparent intestinal permeability from both P3 truncation and the use of the P1′ 4-(trifluoromethyl)phenyl side chain.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-221229 (URN)10.1021/ml400217r (DOI)000333006200005 ()
Available from: 2014-03-26 Created: 2014-03-26 Last updated: 2018-02-04Bibliographically approved
Varasteh, Z., Rosenström, U., Velikyan, I., Mitran, B., Altai, M., Honarvar, H., . . . Orlova, A. (2014). The Effect of Mini-PEG-Based Spacer Length on Binding and Pharmacokinetic Properties of a Ga-68-Labeled NOTA-Conjugated Antagonistic Analog of Bombesin. Molecules, 19(7), 10455-10472
Open this publication in new window or tab >>The Effect of Mini-PEG-Based Spacer Length on Binding and Pharmacokinetic Properties of a Ga-68-Labeled NOTA-Conjugated Antagonistic Analog of Bombesin
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2014 (English)In: Molecules, ISSN 1420-3049, E-ISSN 1420-3049, Vol. 19, no 7, p. 10455-10472Article in journal (Refereed) Published
Abstract [en]

The overexpression of gastrin-releasing peptide receptor (GRPR) in cancer can be used for peptide-receptor mediated radionuclide imaging and therapy. We have previously shown that an antagonist analog of bombesin RM26 conjugated to 1,4,7-triazacyclononane-N, N', N ''-triacetic acid (NOTA) via a diethyleneglycol (PEG(2)) spacer (NOTA-PEG(2)-RM26) and labeled with Ga-68 can be used for imaging of GRPR-expressing tumors. In this study, we evaluated if a variation of mini-PEG spacer length can be used for optimization of targeting properties of the NOTA-conjugated RM26. A series of analogs with different PEG-length (n = 2, 3, 4, 6) was synthesized, radiolabeled and evaluated in vitro and in vivo. The IC50 values of Ga-nat-NOTA-PEG(n)-RM26 (n = 2, 3, 4, 6) were 3.1 +/- 0.2, 3.9 +/- 0.3, 5.4 +/- 0.4 and 5.8 +/- 0.3 nM, respectively. In normal mice all conjugates demonstrated similar biodistribution pattern, however Ga-68-NOTA-PEG(3)-RM26 showed lower liver uptake. Biodistribution of Ga-68-NOTA-PEG(3)-RM26 was evaluated in nude mice bearing PC-3 (prostate cancer) and BT-474 (breast cancer) xenografts. High uptake in tumors (4.6 +/- 0.6% ID/g and 2.8 +/- 0.4% ID/g for PC-3 and BT-474 xenografts, respectively) and high tumor-to-background ratios (tumor/ blood of 44 +/- 12 and 42 +/- 5 for PC-3 and BT-474 xenografts, respectively) were found already at 2 h p.i. of Ga-68-NOTA-PEG(3)-RM26. Results of this study suggest that variation in the length of the PEG spacer can be used for optimization of targeting properties of peptide-chelator conjugates. However, the influence of the mini-PEG length on biodistribution is minor when di-, tri-, tetra- and hexaethylene glycol are compared.

Keywords
bombesin analog, PEG, GRPR, antagonist, molecular imaging, breast cancer, prostate cancer, BT-474, PC-3 cells
National Category
Radiology, Nuclear Medicine and Medical Imaging Cancer and Oncology
Identifiers
urn:nbn:se:uu:diva-231315 (URN)10.3390/molecules190710455 (DOI)000340036200108 ()25036155 (PubMedID)
Available from: 2014-09-08 Created: 2014-09-07 Last updated: 2017-12-05Bibliographically approved
Åberg, O., Varasteh, Z., Lindeberg, G., Larhed, M., Tolmachev, V. & Orlova, A. (2013). (AlF)-F-18-labelling of NOTA-P2-RM26 and its evaluation as a PET ligand for GRPR/BB2. Paper presented at 20th International Symposium on Radiopharmaceutical Sciences; 12-17 May 2013; ICC JEJU, Jeju, Korea. Journal of labelled compounds & radiopharmaceuticals, 56(S1), S404-S404
Open this publication in new window or tab >>(AlF)-F-18-labelling of NOTA-P2-RM26 and its evaluation as a PET ligand for GRPR/BB2
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2013 (English)In: Journal of labelled compounds & radiopharmaceuticals, ISSN 0362-4803, E-ISSN 1099-1344, Vol. 56, no S1, p. S404-S404Article in journal, Meeting abstract (Other academic) Published
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-203006 (URN)000318694100405 ()
Conference
20th International Symposium on Radiopharmaceutical Sciences; 12-17 May 2013; ICC JEJU, Jeju, Korea
Available from: 2013-07-02 Created: 2013-07-01 Last updated: 2017-12-06Bibliographically approved
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