Crystals of Urokinase Type Plasminogen Activator Complexes Reveal the Binding Mode of Peptidomimetic Inhibitors

Żesławska, Ewa; Jacob, Uwe; Schweinitz, Andrea; Coombs, Gary S.; Bode, Wolfram; Madison, Edwin L.

doi:10.1016/s0022-2836(03)00267-5

Cited by 32 publications

(30 citation statements)

References 13 publications

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“…This was expected, because the x-ray structure indicates that the D-Ser hydroxyl group acts as a hydrogen bond donator for the carbonyl oxygen of Leu 97B and as an acceptor for the uPA residue His 99 (15). Surprisingly, only little differences in the uPA inhibition between the D-Ala and D-Dap derivatives were observed, because the free amino group of the D-Dap derivative should still allow the formation of a hydrogen bond to the carbonyl of Leu 97B , which is not possible with the D-Ala compound.…”

Section: Resultsmentioning

confidence: 93%

“…The crystallization of the enzyme-inhibitor complexes, data collection, and structure refinement was performed as described previously using a C122S mutant of the serine protease domain of uPA (␤c-uPA) (15,23). The ␤c-uPA⅐inhibitor complexes were prepared by soaking the inhibitors in crystals of the ␤c-uPA⅐benzamidine complex.…”

Section: Crystallizationmentioning

confidence: 99%

“…For one derivative from this series (Bzls-DSer-Ala-4-Amba) an x-ray structure in complex with uPA was recently published together with the structure of related uPA inhibitors containing a P1-arginal or P1-4-guanidinobenzylamide group (15).…”

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confidence: 99%

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Design of Novel and Selective Inhibitors of Urokinase-type Plasminogen Activator with Improved Pharmacokinetic Properties for Use as Antimetastatic Agents

Schweinitz

Steinmetzer

Banke

et al. 2004

Journal of Biological Chemistry

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124

156

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The serine protease urokinase-type plasminogen activator (uPA) interacts with a specific receptor (uPAR) on the surface of various cell types, including tumor cells, and plays a crucial role in pericellular proteolysis. High levels of uPA and uPAR often correlate with poor prognosis of cancer patients. Therefore, the specific inhibition of uPA with small molecule active-site inhibitors is one strategy to decrease the invasive and metastatic activity of tumor cells. We have developed a series of highly potent and selective uPA inhibitors with a C-terminal 4-amidinobenzylamide residue. Optimization was directed toward reducing the fast elimination from circulation that was observed with initial analogues. The x-ray structures of three inhibitor/uPA complexes have been solved and were used to improve the inhibition efficacy. One of the most potent and selective derivatives, benzylsulfonyl-D-SerSer-4-amidinobenzylamide (inhibitor 26), inhibits uPA with a K i of 20 nM. This inhibitor was used in a fibrosarcoma model in nude mice using lacZ-tagged human HT1080 cells, to prevent experimental lung metastasis formation. Compared with control (100%), an inhibitor dose of 2 ؋ 1.5 mg/kg/day reduced the number of experimental metastases to 4.6 ؎ 1%. Under these conditions inhibitor 26 also significantly prolonged survival. All mice from the control group died within 43 days after tumor cell inoculation, whereas 50% of mice from the inhibitor-treated group survived more than 117 days. This study demonstrates that the specific inhibition of uPA by these inhibitors may be a useful strategy for the treatment of cancer to prevent metastasis.The detachment of malignant cells from the primary tumor and their subsequent migration within the surrounding tissue, including intravasation and extravasation into blood and lymph vessels, leads to tumor dissemination and the formation of metastases at distant loci. Whereas a solid tumor can be removed by surgery or treated by radio-, chemo-, or hormone therapy, invasive tumor cells that have spread over the whole body can form secondary tumors leading to poor prognosis or death of cancer patients. Several proteases, such as matrix metalloproteases, the cysteine proteases cathepsin B and L, the aspartyl protease cathepsin D, and serine proteases, e.g. plasmin and uPA, 1 are involved at multiple stages during growth, invasion and progression of tumors, including metastases formation (1). High levels of expression of these proteases often correlate with poor prognosis for cancer patients (2). However, in several clinical cancer trials with different types of nonspecific matrix metalloprotease inhibitors, disappointing results with poor benefit and severe side effects were observed (3). This stimulated the search for alternative proteases with matrix degrading activity as new targets for anti-cancer drugs. An important role in metastasis has been recently ascribed to the plasmin-plasminogen activation system and especially to uPA.Both, uPA and the second endogenous plasminogen activator tPA are...

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Section: Resultsmentioning

confidence: 93%

Section: Crystallizationmentioning

confidence: 99%

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Design of Novel and Selective Inhibitors of Urokinase-type Plasminogen Activator with Improved Pharmacokinetic Properties for Use as Antimetastatic Agents

Schweinitz

Steinmetzer

Banke

et al. 2004

Journal of Biological Chemistry

Self Cite

124

156

View full text Add to dashboard Cite

show abstract

“…The surface view of chicken ASIC1 (Protein Data Bank code 3HGC) was adapted from Sherwood et al (49), which was originally proposed by the Gouaux group (50,51). The three-dimensional structure of uPA (Protein Data Bank code 1W12) reported by Zeslawska et al was adapted (52). Following removal of the ligand from uPA, docking of the cleavage site in ENaC to uPA was performed with Autodock Vina version 1.1.1 (Scripps Institute, San Diego, CA) in a Pyrx (version 0.85) environment (Scripps Institute).…”

Section: Methodsmentioning

confidence: 99%

Proteolytic Regulation of Epithelial Sodium Channels by Urokinase Plasminogen Activator

Zhao²,

Komissarov³

et al. 2015

Journal of Biological Chemistry

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Background: Depressed fibrinolysis and edema concurrently exist in edematous injury. Results: Divergent regulation of ENaC by urokinase and tPA was observed. Both the catalytic domain of urokinase and cleavage sites in ENaC were identified. Conclusion: Urokinase activates ENaC through catalytic activity-dependent proteolytic modification of ␥ subunit. Significance: Activation of ENaC by urokinase but not tPA provides a novel mechanism for the alleviation of lung edema and pleural effusion.

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“…Up-regulation of uPA/uPAR in tumors and general absence on normal cells make the uPA/uPAR system an attractive target for cancer therapy. Several inhibitors of uPA/uPAR have been designed and tested clinically yielding only limited tumor-static effects (8)(9)(10). Immunotoxins have also been synthesized, which target uPAR.…”

Section: Introductionmentioning

confidence: 99%

Systematic Urokinase-Activated Anthrax Toxin Therapy Produces Regressions of Subcutaneous Human Non–Small Cell Lung Tumor in Athymic Nude Mice

Ortiz

Liu

et al. 2007

Cancer Research

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The novel recombinant anthrax toxin, PrAgU2/FP59, composed of the urokinase-activated protective antigen and a fusion protein of Pseudomonas exotoxin and lethal factor was tested for anti-lung cancer efficacy in an in vivo human tumor model. Male athymic nude mice (age 4-6 weeks) were inoculated s.c. with 10 million H1299 non-small cell lung cancer (NSCLC) cells in the left flank. When tumor volumes reached 200 mm 3 (6-8 days), i.p. injection of 100 ML saline or different ratios and doses of PrAgU2/FP59 in 100 ML saline were given every 3 days for four doses and an additional dose at day 29. Animals were monitored twice daily and tumor measurements were made by calipers. The maximum tolerated doses of PrAgU2/FP59 differed dependent on the ratios of PrAgU2 to FP59 over the range of 3:1 to 25:1, respectively. At tolerated doses, tumor regressions were seen in all animals. Complete histologic remission lasting 60 days occurred in 30% of animals. PrAgU2/FP59 showed dramatic anti-NSCLC efficacy and warrants further clinical development for therapy of patients with advanced NSCLC. [Cancer Res 2007;67(7):3329-36]

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Crystals of Urokinase Type Plasminogen Activator Complexes Reveal the Binding Mode of Peptidomimetic Inhibitors

Cited by 32 publications

References 13 publications

Design of Novel and Selective Inhibitors of Urokinase-type Plasminogen Activator with Improved Pharmacokinetic Properties for Use as Antimetastatic Agents

Design of Novel and Selective Inhibitors of Urokinase-type Plasminogen Activator with Improved Pharmacokinetic Properties for Use as Antimetastatic Agents

Proteolytic Regulation of Epithelial Sodium Channels by Urokinase Plasminogen Activator

Systematic Urokinase-Activated Anthrax Toxin Therapy Produces Regressions of Subcutaneous Human Non–Small Cell Lung Tumor in Athymic Nude Mice

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