Toxicity study of separase inhibitor-Sepin-1 in Sprague-Dawley rats.

Toxicity study of separase inhibitor-Sepin-1 in Sprague-Dawley rats.

Separase activity distribution can be a marker of major molecular response and proliferation of CD34+ cells in TKI-treated chronic myeloid leukemia patients.
Separase activity distribution can be a marker of major molecular response and proliferation of CD34+ cells in TKI-treated chronic myeloid leukemia patients.

Sepin-1 is a small compound that inhibits the enzymatic activity of Separase and growth of cancer cells. As part of the IND-enabling studies to develop Sepin-1 as a chemotherapeutic agent, here we have profiled toxicity Sepin-1 in mice Sprague-Dawley in setting good laboratory practice (GLP). The maximum tolerated dose (MTD) of Sepin-1 in mice is 40 mg / kg in a single dose study and 20 mg / kg dose study for 7 days in a row.

Toxicity study consists of two parts-Main Study and Restoration Study. Sepin-1 to 0 (control), 5 (low dose), 10 (median dose), and 20 (high dose) mg / kg given by bolus intravenous injection to mice once a day for 28 consecutive days. The animals were euthanized at the Main Study Day 29, whereas in the study animals were allowed to recover Recovery for 28 days after 28 days Sepin-1 dose before they were euthanized on Day 29 of the off-dose periods.

Although the effects of Sepin-1 in low and median doses were minimal, hematological analysis shows that high doses of Sepin-1 associated with a decrease in red blood cells and hemoglobin, and an increase in the number of reticulocytes and platelets and mean corpuscular volume. Clinical chemistry showed that Sepin-1 causes increases in total bilirubin and a decrease in creatine kinase. histopathological analysis showed Sepin-1 results in minimal bone marrow erythroid hyperplasia, minimal to moderate hematopoiesis extramedullary spleen, minimal depletion of lymphoid spleen, minimal to mild thymic lymphoid depletion, and minimal to rats mild mandibular lymph nodes of lymphoid hyperplasia in men and women on Main Learn.

Those abnormal changes that Sepin-1 depending on the dose and largely reversible after a 28-day recovery period in animals of Recovery Study. Based on our results, we conclude that Sepin-1 at pharmacological dose (5-10 mg / kg) was well-tolerated, with no significant levels of mortality or morbidity, and can be developed as a potential new drug for treating Separase-expressed tumors.

Toxicity study of separase inhibitor-Sepin-1 in Sprague-Dawley rats.
Toxicity study of separase inhibitor-Sepin-1 in Sprague-Dawley rats.

Cohesin cleavage by separase enhanced with different substrate 

Chromosome segregation began when a cysteine ​​protease, separase, splitting Scc1 transition subunit of cohesin at the metaphase-to-anaphase. Separase inhibited before metaphase with securin tightly bound protein, which contains motifs separase pseudosubstrate blocking the active site.

To investigate the substrate specificity and regulation separase, here we developed a system to produce recombinant human separase securin free. Using these enzymes, we identified motif Scc1 LPE on a substrate different from the cleavage site and necessary for the rapid and specific cleavage of the substrate. LPE Securin also contain conserved motifs, and we provide evidence that this sequence motif involvement Scc1 separase block LPE. Our results indicate that rapid cohesin cleavage by separase require interaction substrate docking outside the active site. This interaction is blocked by securin, providing a second mechanism which inhibits securin cohesin cleavage.


eat-dose pharmacokinetic study Sepin-1 in mice. Sepin-1 intravenously to mice Sprague-Dawley once a day for 28 days in three different (5, 10 and 20mg / kg) dose. Blood samples were taken after dosing on days 1 and 28. Sepin-1 is not stable in phosphate-buffered saline at the base and stable pH buffer such as citric acid-buffered saline (pH 4.0). UHPLC-MS analysis showed Sepin-1 rapidly metabolized in vivo.

Alpha-Synuclein (1.0 mg) Human, Recombinant

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Transthyretin (V30G) (1.0 mg) Human, Recombinant

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Transthyretin (T119M) (1.0 mg) Human, Recombinant

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Transthyretin (T60A) (1.0mg) Human, Recombinant

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Transthyretin (A97S) (1.0mg) Human, Recombinant

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Transthyretin (S52P) (1.0 mg) Human, Recombinant

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Transthyretin (V30I) (1.0 mg) Human, Recombinant

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ELISA Flex: Bovine TNF-alpha (ALP)

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ELISA Flex: Bovine TNF-alpha (HRP)

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ELISA Flex: Bovine TNF-alpha (HRP)

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EUR 351.75

ELISpot Flex: Bovine TNF-alpha (ALP)

3012-2A for 4 plates
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ELISpot Flex: Bovine TNF-alpha (HRP)

3012-2H for 4 plates
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Anti-bovine TNF-alpha mAb (MTF4918), unconjugated

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Anti-bovine TNF-alpha mAb (MTF4918), unconjugated

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ELISA Flex: Porcine IL-17A (ALP)

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EUR 351.75

ELISA Flex: Porcine IL-17A (HRP)

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ELISA Flex: Porcine IL-17A (HRP)

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ELISpot Flex: Porcine IL-17A (ALP)

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Anti-woodchuck IFN-gamma mAb (MT225), biotin

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ELISA Flex: Hamster IFN-gamma (ALP)

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ELISA Flex: Hamster IFN-gamma (ALP)

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ELISA Flex: Hamster IFN-gamma (HRP)

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ELISA Flex: Hamster IFN-gamma (HRP)

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EUR 351.75

ELISpot Flex: Hamster IFN-gamma (ALP)

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ELISpot Flex: Hamster IFN-gamma (HRP)

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Anti-hamster IFN-gamma mAb (MTH21), unconjugated

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Anti-hamster IFN-gamma mAb (MTH21), unconjugated

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ELISpot Plus: Hamster IFN-gamma (ALP)

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ELISpot Plus: Hamster IFN-gamma (ALP)

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ELISpot Plus: Hamster IFN-gamma (HRP)

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ELISpot Plus: Hamster IFN-gamma (HRP)

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EUR 535.5

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Anti-hamster IFN-gamma mAb (MTH29), biotin

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Anti-ferret IL-2 mAb (MT264), unconjugated

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ELISA Flex: Ferret IL-2 (ALP)

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ELISA Flex: Ferret IL-2 (ALP)

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EUR 351.75

ELISA Flex: Ferret IL-2 (HRP)

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ELISA Flex: Ferret IL-2 (HRP)

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EUR 351.75

ELISpot Flex: Ferret IL-2 (ALP)

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EUR 609

ELISpot Flex: Ferret IL-2 (HRP)

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Anti-ferret IL-2 mAbs (MT264/271), unconjugated

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Anti-ferret IL-2 mAbs (MT264/271), unconjugated

3104-3-250 250 µg
EUR 241.5

ELISA Flex: Bovine IL-17A (ALP)

3109-1A-20 for 20 plates
EUR 651

ELISA Flex: Bovine IL-17A (ALP)

3109-1A-6 for 6 plates
EUR 351.75

ELISA Flex: Bovine IL-17A (HRP)

3109-1H-20 for 20 plates
EUR 651

ELISA Flex: Bovine IL-17A (HRP)

3109-1H-6 for 6 plates
EUR 351.75

ELISpot Flex: Bovine IL-17A (ALP)

3109-2A for 4 plates
EUR 609

ELISpot Flex: Bovine IL-17A (HRP)

3109-2H for 4 plates
EUR 609

Anti-bovine IL-17A mAb (MT49A7), unconjugated

3109-3-1000 1 mg
EUR 840

Anti-bovine IL-17A mAb (MT49A7), unconjugated

3109-3-250 250 µg
EUR 241.5

ELISpot Plus: Bovine IL-17A (ALP)

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EUR 2420.25

ELISpot Plus: Bovine IL-17A (ALP)

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EUR 535.5

ELISpot Plus: Bovine IL-17A (HRP)

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EUR 2420.25

ELISpot Plus: Bovine IL-17A (HRP)

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EUR 535.5

Anti-bovine IL-17A mAb (MT51B8), biotin

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Anti-bovine IL-17A mAb (MT51B8), biotin

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EUR 283.5

ELISA Flex: Rabbit IFN-gamma (ALP)

3110-1A-20 for 20 plates
EUR 651

ELISA Flex: Rabbit IFN-gamma (ALP)

3110-1A-6 for 6 plates
EUR 351.75

ELISA Flex: Rabbit IFN-gamma (HRP)

3110-1H-20 for 20 plates
EUR 651

ELISA Flex: Rabbit IFN-gamma (HRP)

3110-1H-6 for 6 plates
EUR 351.75

ELISpot Flex: Rabbit IFN-gamma (ALP)

3110-2A for 4 plates
EUR 609

ELISpot Flex: Rabbit IFN-gamma (HRP)

3110-2H for 4 plates
EUR 609

Anti-rabbit IFN-gamma mAb (MT327), unconjugated

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Anti-rabbit IFN-gamma mAb (MT327), unconjugated

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EUR 241.5

ELISpot Plus: Rabbit IFN-gamma (ALP)

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EUR 2420.25

ELISpot Plus: Rabbit IFN-gamma (ALP)

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EUR 535.5

ELISpot Plus: Rabbit IFN-gamma (HRP)

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EUR 2420.25

ELISpot Plus: Rabbit IFN-gamma (HRP)

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EUR 535.5

Anti-rabbit IFN-gamma mAb (MT318), biotin

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Anti-rabbit IFN-gamma mAb (MT318), biotin

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Anti-rabbit IFN-gamma mAb (MT318), PF647P

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ELISA Flex: Bovine IL-2 (ALP)

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ELISA Flex: Bovine IL-2 (ALP)

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EUR 351.75

ELISA Flex: Bovine IL-2 (HRP)

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EUR 651

ELISA Flex: Bovine IL-2 (HRP)

3111-1H-6 for 6 plates
EUR 351.75

ELISpot Flex: Bovine IL-2 (ALP)

3111-2A for 4 plates
EUR 609

ELISpot Flex: Bovine IL-2 (HRP)

3111-2H for 4 plates
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Anti-bovine IL-2 mAb (MT11A31), unconjugated

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Anti-bovine IL-2 mAb (MT11A31), unconjugated

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EUR 241.5

ELISpot Plus: Bovine IL-2 (ALP)

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ELISpot Plus: Bovine IL-2 (ALP)

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ELISpot Plus: Bovine IL-2 (HRP)

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EUR 2420.25

ELISpot Plus: Bovine IL-2 (HRP)

3111-4HPW-2 2 plates
EUR 535.5

Anti-bovine IL-2 mAb (MT3B3), unconjugated

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EUR 241.5

Anti-bovine IL-2 mAb (MT3B3), biotin

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Anti-bovine IL-2 mAb (MT3B3), PF488P

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EUR 304.5

Anti-bovine IL-2 mAb (MT3B3), PF647P

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ELISA Flex: Ferret IFN-gamma (ALP)

3112-1A-20 for 20 plates
EUR 651

ELISA Flex: Ferret IFN-gamma (ALP)

3112-1A-6 for 6 plates
EUR 351.75

ELISA Flex: Ferret IFN-gamma (HRP)

3112-1H-20 for 20 plates
EUR 651

ELISA Flex: Ferret IFN-gamma (HRP)

3112-1H-6 for 6 plates
EUR 351.75

ELISpot Flex: Ferret IFN-gamma (ALP)

3112-2A for 4 plates
EUR 609

ELISpot Flex: Ferret IFN-gamma (HRP)

3112-2H for 4 plates
EUR 609

Anti-ferret IFN-gamma mAb (MTF14), unconjugated

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Anti-ferret IFN-gamma mAb (MTF14), unconjugated

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EUR 241.5

ELISpot Plus: Ferret IFN-gamma (ALP)

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EUR 2420.25

ELISpot Plus: Ferret IFN-gamma (ALP)

3112-4APW-2 2 plates
EUR 535.5

ELISpot Plus: Ferret IFN-gamma (HRP)

3112-4HPW-10 10 plates
EUR 2420.25

ELISpot Plus: Ferret IFN-gamma (HRP)

3112-4HPW-2 2 plates
EUR 535.5

Anti-ferret IFN-gamma mAb (MTF19), biotin

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EUR 1044.75

Anti-ferret IFN-gamma mAb (MTF19), biotin

3112-6-250 250 µg
EUR 283.5

Anti-ferret IFN-gamma mAb (MTF14), PF488P

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EUR 304.5

Anti-ferret IFN-gamma mAb (MTF14), PF647P

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EUR 304.5

ELISA Flex: Canine IFN-gamma (ALP)

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EUR 651

ELISA Flex: Canine IFN-gamma (ALP)

3113-1A-6 for 6 plates
EUR 351.75

ELISA Flex: Canine IFN-gamma (HRP)

3113-1H-20 for 20 plates
EUR 651

ELISA Flex: Canine IFN-gamma (HRP)

3113-1H-6 for 6 plates
EUR 351.75

ELISpot Flex: Canine IFN-gamma (ALP)

3113-2A for 4 plates
EUR 609

ELISpot Flex: Canine IFN-gamma (HRP)

3113-2H for 4 plates
EUR 609

Anti-canine IFN-gamma mAb (MT13), unconjugated

3113-3-1000 1 mg
EUR 840

Anti-canine IFN-gamma mAb (MT13), unconjugated

3113-3-250 250 µg
EUR 241.5

ELISpot Plus: Canine IFN-gamma (ALP)

3113-4APW-10 10 plates
EUR 2420.25

ELISpot Plus: Canine IFN-gamma (ALP)

3113-4APW-2 2 plates
EUR 535.5

ELISpot Plus: Canine IFN-gamma (HRP)

3113-4HPW-10 10 plates
EUR 2420.25

ELISpot Plus: Canine IFN-gamma (HRP)

3113-4HPW-2 2 plates
EUR 535.5

Anti-canine IFN-gamma mAb (MT166), biotin

3113-6-1000 1 mg
EUR 1044.75

Anti-canine IFN-gamma mAb (MT166), biotin

3113-6-250 250 µg
EUR 283.5

Anti-bovine IL-8 (CXCL8) mAbs (MT8H6/8F19), unconjugated

3114-1-1000 1 mg
EUR 840

Anti-bovine IL-8 (CXCL8) mAbs (MT8H6/8F19), unconjugated

3114-1-250 250 µg
EUR 241.5

ELISA Flex: Bovine IL-8 (ALP)

3114-1A-20 for 20 plates
EUR 651

ELISA Flex: Bovine IL-8 (ALP)

3114-1A-6 for 6 plates
EUR 351.75

ELISA Flex: Bovine IL-8 (HRP)

3114-1H-20 for 20 plates
EUR 651

ELISA Flex: Bovine IL-8 (HRP)

3114-1H-6 for 6 plates
EUR 351.75

ELISpot Flex: Bovine IL-8 (ALP)

3114-2A for 4 plates
EUR 609

ELISpot Flex: Bovine IL-8 (HRP)

3114-2H for 4 plates
EUR 609

Anti-bovine IL-8 (CXCL8) mAb (MT8H6), unconjugated

3114-3-1000 1 mg
EUR 840

Anti-bovine IL-8 (CXCL8) mAb (MT8H6), unconjugated

3114-3-250 250 µg
EUR 241.5

ELISpot Plus: Bovine IL-8 (ALP)

3114-4APW-10 10 plates
EUR 2420.25

One of the major metabolite is stirring the amine of 2,2-dimethyl-5-nitro-2H-benzimidazole (named Sepin-1.55). Sepin-concentration in the blood samples was 1.55 Sepin-1 depending on the dose and used for pharmacokinetic analysis Sepin-1. Tmax was about 5-15 minutes.

Karina

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