Screening of a Library for Factor VIIa Inhibitors

2021 ◽  
Vol 18 ◽  
Author(s):  
Alexandre Bridoux ◽  
Shaker A. Mousa
Keyword(s):  
Small Molecule ◽  
Factor Viia ◽  
Chorioallantoic Membrane ◽  
High Technology ◽  
Coagulation Cascade ◽  
Chemical Structures ◽  
Pathological Angiogenesis ◽  
Chick Chorioallantoic Membrane ◽  
Cam Model

Background: As an alternative to the anticoagulant’s strategy using direct or indirect anti-Xa drugs, considering other targets upstream in the coagulation cascade such as anti-Factor VIIa could represent an effective and safer strategy in coagulation and pathological angiogenesis. Objective: The objective of the study was to assess a high technology methodology composed of virtual screening, anticoagulant, and anti-angiogenesis assays to identify potent small-molecule FVIIa inhibitors. Methods: Chemical databanks were screened to select molecules bearing functional groups that could fit into the active site of FVIIa, which were then tested. Ligands assigned with the lowest scores were retained and then biologically assessed. Results: From the 500 molecules considered, 8 chemical structures revealed to be effective compounds in vitro and to inhibit angiogenesis in the chick chorioallantoic membrane (CAM) model. Conclusion: New potent small-molecule FVIIa inhibitors have been identified; further biochemical and chemical developments would be investigated directly from the selected scaffolds.

Sulfur- and Selenium-Containing Compounds Derived from Natural and Synthetic Sources in Angiogenesis Modulation.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3924-3924 ◽  
Author(s):  
Shaker A. Mousa ◽  
Seema Mohamed ◽  
Laura O’Connor ◽  
Eric Block
Keyword(s):  
Growth Factor ◽  
Chorioallantoic Membrane ◽  
Tube Formation ◽  
Therapeutic Effects ◽  
Fibroblast Growth ◽  
Chick Chorioallantoic Membrane ◽  
Cam Model ◽  
Sulfur Containing ◽  
Sulfur Containing Compounds

Abstract Increases in serum sulfate may explain some of the therapeutic effects of methylsulfonyl methane (MSM), dimethyl sulfoxide, and glucosamine sulfate. Organic sulfur, as sulfur-containing amino acids, can be used to increase synthesis of S-adenosylmethionine, glutathione, taurine, and N-acetylcysteine. MSM may be effective for the treatment of allergy, pain syndromes, athletic injuries, and bladder disorders. Epidemiological studies have suggested that brassica vegetables are protective against cancers of the lungs and alimentary tract. Cruciferous vegetables are the dietary source of glucosinolates, a large group of sulfur-containing glycosides. However, dosages, mechanisms of action, and rationales for the use of various sulfur-containing compounds from natural or food products need to be better defined. The low toxicological profiles of these sulfur compounds, combined with promising therapeutic effects, warrant continued preclinical and clinical investigations. In this current study the angiostatic (antiangiogenesis) efficacy for individual sulfur-containing compounds derived from natural or synthetic sources was determined using in vitro (endothelial cell [EC] tube formation assay) and in vivo (chick chorioallantoic membrane [CAM] model). In vitro EC tube formation studies demonstrated significant inhibition of fibroblast growth factor-2 (FGF-2) stimulated EC tube formation by the different sulfur- and selenium-containing compounds, with different potency depending on the oxidation state of the sulfur or selenium. Furthermore, utilizing the CAM model, similar inhibitory efficacy of FGF-2 induced angiogenesis. These data suggest that sulfur and selenium compounds derived from natural sources might be a useful therapy for the inhibition of angiogenesis associated with human tumor growth and other pathological angiogenesis-mediated disorders, such as ocular and inflammatory diseases. Antiangiogenesis Efficacy of Sulfone/Sulfoxide and Selenone/Selenoxide in the Chick Chorioallantoic Membrane Model Treatment Branch Points ± SEM % Inhibition ± SEM FGF = fibroblast growth factor; PBS = phosphate-buffered saline. PBS control 64.67 ± 3.00 ----- FGF-2 (1.5 ug/ml) 178.20 ± 5.32 ----- FGF-2 + diphenyl sulfoxide (100 uM) 145.65 ± 11.10 34.12 ± 4.8 FGF-2 + diphenyl sulfone (3 uM) 82.56 ± 4.4 84.25 ± 3.6 FGF-2 + dimethyl selenoxide (30 uM) 120.86 ± 3.1 50.74 ± 2.7 FGF-2 + dimethyl selenone (3 uM) 71.63 ± 3.8 93.87 ± 3.3

scholarly journals Management of the Bleeding Patient Receiving New Oral Anticoagulants: A Role for Prothrombin Complex Concentrates

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Lisa M. Baumann Kreuziger ◽  
Joseph C. Keenan ◽  
Colleen T. Morton ◽  
David J. Dries
Keyword(s):  
Recombinant Factor Viia ◽  
Factor Viia ◽  
Oral Anticoagulants ◽  
New Oral Anticoagulants ◽  
Coagulation Cascade ◽  
Clotting Factors ◽  
Prothrombin Complex Concentrates ◽  
Clotting System ◽  
Clinical Trial Evidence

Ease of dosing and simplicity of monitoring make new oral anticoagulants an attractive therapy in a growing range of clinical conditions. However, newer oral anticoagulants interact with the coagulation cascade in different ways than traditional warfarin therapy. Replacement of clotting factors will not reverse the effects of dabigatran, rivaroxaban, or apixaban. Currently, antidotes for these drugs are not widely available. Fortunately, withholding the anticoagulant and dialysis are freqnently effective treatments, particularly with rivaroxaban and dabigatran. Emergent bleeding, however, requires utilization of Prothrombin Complex Concentrates (PCCs). PCCs, in addition to recombinant factor VIIa, are used to activate the clotting system to reverse the effects of the new oral anticoagulants. In cases of refractory or emergent bleeding, the recommended factor concentrate in our protocols differs between the new oral anticoagulants. In patients taking dabigatran, we administer an activated PCC (aPCC) [FELBA] due to reported benefit in human in vitro studies. Based on human clinical trial evidence, the 4-factor PCC (Kcentra) is suggested for patients with refractory rivaroxaban- or apixaban-associated hemorrhage. If bleeding continues, recombinant factor VIIa may be employed. With all of these new procoagulant agents, the risk of thrombosis associated with administration of factor concentrates must be weighed against the relative risk of hemorrhage.

scholarly journals The growth in vitro of vaccinia virus in chick embryo chorio-allantoic memberane, minced embryo and cell suspensions

1957 ◽  
Vol 55 (3) ◽  
pp. 347-360 ◽  
Author(s):  
H. B. Maitland ◽  
D. I. Magrath
Keyword(s):  
Chick Embryo ◽  
Vaccinia Virus ◽  
Growth Curve ◽  
Chorioallantoic Membrane ◽  
Growth Cycle ◽  
Cell Suspensions ◽  
Rabbit Skin ◽  
Chick Chorioallantoic Membrane ◽  
Considerable Doubt

The growth curve of rabbit skin-adapted vaccinia virus in the chick chorioallantoic membrane incubated in Hanks' solution showed a drop in titre of virus for about 10 hr. followed by growth. At least 25% of virus, sometimes more, remained infective. A similar fall in titre was observed in heated membranes in which the virus did not grow and this occurred also when membranes, either normal or heated, were infected and disintegrated before incubation.The growth curve of virus in minced chick-embryo was similar to that in chorioallantoic membrane.Virus in cell suspensions prepared from chick embryo and incubated in a nutrient medium showed only a small loss of infectivity before growth in some experiments and rarely dropped below 65–70 % of the original titre in others.These results throw considerable doubt on the view that loss of infectivity preceding growth of vaccinia virus should be interpreted as an essential part of a growth cycle.

The chick embryo chorioallantoic membrane model: a research approach for ex vivo and in vivo experiments

2021 ◽  
Vol 28 ◽  
Author(s):  
Ana Isabel Fraguas-Sánchez ◽  
Cristina Martín-Sabroso ◽  
Ana Isabel Torres-Suárez
Keyword(s):  
Animal Models ◽  
Chorioallantoic Membrane ◽  
New Drugs ◽  
Biological Research ◽  
Research Areas ◽  
Chick Chorioallantoic Membrane ◽  
Biodistribution Studies ◽  
Toxicological Studies ◽  
Cam Model

Background: The chick chorioallantoic membrane (CAM) model has attracted a great deal of interest in pharmaceutical and biological research as an alternative or complementary in vivo assay to animal models. Traditionally, CAM assay has been widely used to perform some toxicological studies, specifically to evaluate the skin, ocular and embryo toxicity of new drugs and formulations, and perform angiogenesis studies. Due to the possibility to generate the tumors onto the CAM, this model has also become an excellent strategy to evaluate the metastatic potential of different tumours and test the efficacy of novel anticancer therapies in vivo. Moreover, in the recent years, its use has considerably grown in other research areas, including the evaluation of new anti-infective agents, the development of biodistribution studies and tissue engineering research. Objectives: This manuscript provides a critical overview of the use of CAM model in pharmaceutical and biological research, especially to test the toxicity of new drugs and formulations and the biodistribution and the efficacy of novel anticancer and anti-infective therapies, analyzing its advantages and disadvantages compared to animal models. Conclusion: The chick chorioallantoic membrane model shows great utility in several research areas, such as cancer, toxicology, biodistribution studies and anti-infective therapies. In fact, it has become an intermediate stage between in vitro experiments and animal studies, and, in the case of toxicological studies (skin and ocular toxicity), has even replaced the animal models.

scholarly journals A novel method of screening combinations of angiostatics identifies bevacizumab and temsirolimus as synergistic inhibitors of glioma-induced angiogenesis

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252233
Author(s):  
Michael I. Dorrell ◽  
Heidi R. Kast-Woelbern ◽  
Ryan T. Botts ◽  
Stephen A. Bravo ◽  
Jacob R. Tremblay ◽  
...  
Keyword(s):  
Side Effects ◽  
Tumor Angiogenesis ◽  
Clinical Success ◽  
Low Cost ◽  
Chorioallantoic Membrane ◽  
Cellular Interactions ◽  
Compensatory Mechanisms ◽  
Chick Chorioallantoic Membrane

Tumor angiogenesis is critical for the growth and progression of cancer. As such, angiostasis is a treatment modality for cancer with potential utility for multiple types of cancer and fewer side effects. However, clinical success of angiostatic monotherapies has been moderate, at best, causing angiostatic treatments to lose their early luster. Previous studies demonstrated compensatory mechanisms that drive tumor vascularization despite the use of angiostatic monotherapies, as well as the potential for combination angiostatic therapies to overcome these compensatory mechanisms. We screened clinically approved angiostatics to identify specific combinations that confer potent inhibition of tumor-induced angiogenesis. We used a novel modification of the ex ovo chick chorioallantoic membrane (CAM) model that combined confocal and automated analyses to quantify tumor angiogenesis induced by glioblastoma tumor onplants. This model is advantageous due to its low cost and moderate throughput capabilities, while maintaining complex in vivo cellular interactions that are difficult to replicate in vitro. After screening multiple combinations, we determined that glioblastoma-induced angiogenesis was significantly reduced using a combination of bevacizumab (Avastin®) and temsirolimus (Torisel®) at doses below those where neither monotherapy demonstrated activity. These preliminary results were verified extensively, with this combination therapy effective even at concentrations further reduced 10-fold with a CI value of 2.42E-5, demonstrating high levels of synergy. Thus, combining bevacizumab and temsirolimus has great potential to increase the efficacy of angiostatic therapy and lower required dosing for improved clinical success and reduced side effects in glioblastoma patients.

scholarly journals Mobilization of shell calcium by the chick chorioallantoic membrane in vitro.

1994 ◽  
Vol 190 (1) ◽  
pp. 141-153
Author(s):  
M J Packard
Keyword(s):  
Domestic Fowl ◽  
Culture Medium ◽  
Chorioallantoic Membrane ◽  
The Other ◽  
Chick Chorioallantoic Membrane ◽  
Petri Dishes ◽  
Specific Degradation ◽  
Donor Eggs

Two explants of shell were removed from each of several fertile eggs of domestic fowl at different times during incubation. The chorioallantoic membrane (CAM) was removed from one of the explants (SHELL ONLY) and was left in situ on the other (SHELL+CAM). Explants were cultured for 24, 48 or 96 h at 37 degrees C and 5% CO2 in air in individual Petri dishes containing Dulbecco's modified Eagle's medium, bovine serum albumin, penicillin and streptomycin. Both SHELL+CAM and SHELL ONLY explants released calcium into the culture medium, but the former released considerably more calcium than the latter. More calcium was released by SHELL+CAM explants taken from older eggs than from younger ones, but the age of the donor eggs did not affect release of calcium by SHELL ONLY explants. In addition, release of calcium by SHELL+CAM explants exceeded that shown by SHELL ONLY explants for multiple 24 h intervals. However, the capacity for sustained release of calcium by SHELL+CAM explants declined with age and maturity of the CAM. Manipulations that lead to the death of the CAM abolish the capacity for SHELL+CAM explants to release more calcium than SHELL ONLY explants. Differential release of calcium by SHELL+CAM explants was not attributable to calcium present in the CAM at the onset of culture or to non-specific degradation of the shell by intracellular constituents released as a result of the death of the CAM. Taken in concert, these results indicate that the CAM mobilizes calcium from the eggshell during in vitro culture.

scholarly journals Angiogenic property of silk fibroin scaffolds with adipose-derived stem cells on chick chorioallantoic membrane

2021 ◽  
Vol 8 (3) ◽  
Author(s):  
Tanapong Watchararot ◽  
Weerapong Prasongchean ◽  
Peerapat Thongnuek
Keyword(s):  
Stem Cells ◽  
Silk Fibroin ◽  
Pore Diameter ◽  
Chorioallantoic Membrane ◽  
Control Group ◽  
Adipose Derived Stem Cells ◽  
Chick Chorioallantoic Membrane ◽  
Human Adipose Stem Cells

Angiogenesis is a crucial step in tissue regeneration and repair. Biomaterials that allow or promote angiogenesis are thus beneficial. In this study, angiogenic properties of salt-leached silk fibroin (SF) scaffolds seeded with human adipose stem cells (hADSCs) were studied using chick chorioallantoic membrane (CAM) as a model. The hADSC-seeded SF scaffolds (SF-hADSC) with the porosity of 77.34 ± 6.96% and the pore diameter of 513.95 ± 4.99 µm were implanted on the CAM of chick embryos that were on an embryonic day 8 (E8) of development. The SF-hADSC scaffolds induced a spoke-wheel pattern of capillary network indicative of angiogenesis, which was evident since E11. Moreover, the ingrowth of blood vessels into the scaffolds was seen in histological sections. The unseeded scaffolds induced the same extent of angiogenesis later on E14. By contrast, the control group could not induce the same extent of angiogenesis. In vitro cytotoxicity tests and in vivo angioirritative study reaffirmed the biocompatibility of the scaffolds. This work highlighted that the biocompatible SF-hADSC scaffolds accelerate angiogenesis, and hence they can be a promising biomaterial for the regeneration of tissues that require angiogenesis.

Transepithelial calcium transport in the chick chorioallantoic membrane. I. Isolation and characterization of chorionic ectoderm cells

1993 ◽  
Vol 105 (2) ◽  
pp. 369-379 ◽  
Author(s):  
R.E. Akins ◽  
R.S. Tuan
Keyword(s):  
Calcium Binding ◽  
Chorioallantoic Membrane ◽  
Experimental Manipulation ◽  
Transepithelial Transport ◽  
Temperature Sensitive ◽  
Specific Cell ◽  
Chick Chorioallantoic Membrane ◽  
Isolation And Characterization

The chicken eggshell supplies approximately 80% of the calcium found in the hatchling chick. The mobilization of eggshell calcium into the developing embryo involves the transepithelial transport of large amounts of calcium in a development-specific manner. The cells responsible for the transport of eggshell calcium into the embryonic circulation are the ectodermal cells of the chorioallantoic membrane. In this report, we present a method for the isolation and culture of chorioallantoic membrane ectodermal cells, which are amenable to direct experimental manipulation. Cell preparations are characterized with respect to the expression of an ectoderm-specific cell surface marker (transcalcin, a calcium-binding protein), and a specific enzymatic activity (elevated Ca(2+)-activated ATPase). Functional assessment of in vitro cellular calcium uptake by 45Ca2+ tracer kinetics indicates the persistence of a temperature-sensitive, rapid-influx pathway similar to that observed in vivo. The preparations of primary ectodermal cells present an in vitro system applicable to the experimental analysis of calcium metabolism and transport by the chick chorioallantoic membrane.

Sign in / Sign up

Export Citation Format

Share Document