Pharmacokinetics and pharmacodynamics of a recombinant fusion protein linking activated coagulation factor VII with human albumin (rVIIa-FP) in patients with congenital FVII deficiency

Abstract A recombinant fusion protein linking coagulation factor IX (FIX) with human albumin (rIX-FP) has been developed to facilitate hemophilia B treatment by less frequent FIX dosing. This first-in-human dose-escalation trial in 25 previously treated subjects with hemophilia B (FIX ≤ 2 IU/dL) examined the safety and pharmacokinetics of 25, 50, and 75 IU/kg rIX-FP. Patients in the 50-IU/kg cohort underwent a comparative pharmacokinetics assessment with their previous FIX product (plasma-derived or recombinant). No allergic reactions or inhibitors were observed. Four mild, possibly treatment-related adverse events were reported. In the 50-IU/kg cohort (13 subjects), the mean half-life of rIX-FP was 92 hours, more than 5 times longer than the subjects' previous FIX product. After 25 or 50 IU/kg rIX-FP administration, the baseline-corrected mean FIX activity remained elevated at day 7 (7.4 IU/dL and 13.4 IU/dL, respectively) and day 14 (2.5 IU/dL and 5.5 IU/dL, respectively). The incremental recovery of rIX-FP was higher than both recombinant and plasma-derived FIX (1.4 vs 0.95 and 1.1 IU/dL per IU/kg, respectively). These results demonstrated both the safety and improved pharmacokinetics of rIX-FP, thus indicating this new product with extended half-life as possibly able to control and prevent bleeding with less frequent injection. The trial was registered at www.clinicaltrials.gov as no. NCT01233440.

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Monitoring of Recombinant Activated Coagulation Factor VII (rFVIIa) Therapy in Patients (pts) with Inherited Factor VII (FVII) Deficiency

Blood ◽

10.1182/blood-2019-124709 ◽

2019 ◽

Vol 134 (Supplement_1) ◽

pp. 2412-2412

Author(s):

Gennadii M. Galstian ◽

Olesya A. Polevodova ◽

Elena Yakovleva ◽

Antonina Shchekina ◽

Igor Davydkin ◽

...

Keyword(s):

Conflicts Of Interest ◽

Coagulation Factor ◽

Screening Tests ◽

Factor Vii ◽

Extrinsic Pathway ◽

Coagulation Factor Vii ◽

Hemostatic Effect ◽

Normal Ranges ◽

Rfviia Administration ◽

Fvii Deficiency

Introduction and Objectives: Inherited FVII deficiency is a rare hemorrhagic disorder.Clinical bleeding does not correlate with the level of FVII plasma activity. FVII involves in the extrinsic coagulation pathway. ROTEM is point of care method. EXTEM is one of ROTEM screening tests. EXTEM analyses the extrinsic pathway of coagulation. Aim of the study was to evaluate hemostatic effect and safety of rFVIIa treatment in pts with inherited FVII deficiency. Materials and Methods: The results of rFVIIa treatment were investigated in 4 pts (1 male, 3 females) with inherited FVII deficiency. Pts were treated with rFVIIa before gynecological (2), abdominal (1) and orthopedic surgery (1). Pts received 30 mcg/kg of rFVIIa (Koagil VII, Generium, Russia) before surgery. Plasma FVII activity, fibrinogen, INR, APTT, ROTEM and thromboelastography (TEG) parameters were investigated before rFVIIa administration, then in 15 minutes, in 2 hours, in 6 hours and in 12 hours. Results: Before rFVIIa administration median plasma FVII activity was low, CT EXTEM was long and INR was greater than 3. TEG, APTT and fibrinogen were in normal ranges (table 1, fig. 1). Fifteen minutes after rFVIIa administration FVII:C activity increased, INR and CT EXTEM decreased. Hemostatic effect of rFVIIa remained during 12 hours. During this period CT EXTEM correlated with INR (r = 0.97, p<0.001). Other parameters did not change significantly. None patients had hemorrhagic complications. Conclusion: CT EXTEM allowed to evaluate hemostatic effect of rFVIIa in operating room and good correlated with INR in pts with inherited FVII deficiency. Disclosures No relevant conflicts of interest to declare.

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Molecular characterization of iranian patients with inherited coagulation factor VII deficiency

Balkan Journal of Medical Genetics ◽

10.1515/bjmg-2017-0027 ◽

2017 ◽

Vol 20 (2) ◽

pp. 19-25

Author(s):

S Shahbazi ◽

R Mahdian ◽

K Karimi ◽

A Mashayekhi

Keyword(s):

Coagulation Factor ◽

Compound Heterozygote ◽

Phenotypic Heterogeneity ◽

Coagulation Cascade ◽

Factor Vii ◽

Missense Mutations ◽

Protein Activity ◽

Coagulation Factor Vii ◽

Fvii Deficiency ◽

Iranian Patients

AbstractCoagulation factor VII (FVII) is a key enzyme of the extrinsic coagulation cascade that is predominantly produced by hepatocytes. TheF7gene mutations cause FVII deficiency with considerable molecular and phenotypic heterogeneity. We characterized the molecular alterations of theF7gene and their corresponding mRNA transcripts in Iranian patients from eight unrelated families. The mutations were detected by polymerase chain reaction (PCR)-sequencing of allF7gene exons, their flanking intronic sequences, as well as their corresponding cDNA fragments. Homozygous P303T, C91S and R304Q mutations were detected in patient 2, patient 5, and patient 6, respectively. Patient 7 was a compound heterozygote for S282R and H348R and patient 8 was a compound heterozygote for R304Q and IVS7+7A>G mutations. Furthermore, our investigation revealed three heterozygous individuals, patient 1 and patient 3 with the A244V mutation who were symptomatic and patient 4 with V(–39)I mutation who was also asymptomatic. TheF7mRNA expression analysis revealed that, except the transcript of V(–39)I, other mutation-harboring transcripts were expressed at detectable levels. In conclusion, this report reinforces the genetic and phenotypic heterogeneity of FVII deficiency. The findings of the mRNA study implied that decreased FVII protein activity subsequent to missense mutations does not completely reflect the degradation of mutation-harboring mRNA.

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Exclusion of the First EGF Domain of Factor VII by a Splice Site Mutation Causes Lethal Factor VII Deficiency

Blood ◽

10.1182/blood.v92.3.920.415a18_920_926 ◽

1998 ◽

Vol 92 (3) ◽

pp. 920-926

Author(s):

John H. McVey ◽

Emma J. Boswell ◽

Osamu Takamiya ◽

Gabriel Tamagnini ◽

Victor Valente ◽

...

Keyword(s):

Splice Site ◽

Lethal Factor ◽

Coagulation Factor ◽

Factor Vii ◽

Single Point Mutation ◽

Severe Bleeding ◽

Polymorphism Analysis ◽

Nucleotide Position ◽

Coagulation Factor Vii ◽

Fvii Deficiency

We have studied a family with homozygous lethal, blood coagulation factor VII (FVII) deficiency. To identify the mutation responsible for the deficiency, exons 2 to 8 and the intron-exon junctions of their FVII genes were amplified from peripheral white blood cell DNA by polymerase chain reaction and screened by single-strand conformational polymorphism analysis. The fragment showing aberrant mobility was cloned and sequenced. We detected a single point mutation, a homozygous G to A substitution at nucleotide position 6070, in the invariant GT dinucleotide at the 5′ splice site of intron 4. Homozygosity was confirmed by loss of a site for the restriction endonuclease Mlu I. Analysis of the splicing pattern of ectopic transcripts in lymphocytes in the parents revealed that this mutation is associated with skipping of exon 4, which produces an mRNA encoding FVII with an in-frame deletion of the first epidermal growth factor–like domain (EGF 1). Transient transfection of COS-7 cells with an expression vector containing the ▵EGF 1 FVII cDNA shows that this mutant protein is not expressed. The identification of the molecular basis of the FVII deficiency in this family allowed mutation-specific prenatal diagnosis to be performed in a subsequent pregnancy. In this family complete FVII deficiency is associated with a severe bleeding diathesis but no developmental abnormalities, lending weight to the hypothesis that fetal FVII is not required for the putative angiogenic functions of tissue factor in humans. © 1998 by The American Society of Hematology.

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Exclusion of the First EGF Domain of Factor VII by a Splice Site Mutation Causes Lethal Factor VII Deficiency

Blood ◽

10.1182/blood.v92.3.920 ◽

1998 ◽

Vol 92 (3) ◽

pp. 920-926 ◽

Cited By ~ 33

Author(s):

John H. McVey ◽

Emma J. Boswell ◽

Osamu Takamiya ◽

Gabriel Tamagnini ◽

Victor Valente ◽

...

Keyword(s):

Splice Site ◽

Lethal Factor ◽

Coagulation Factor ◽

Factor Vii ◽

Single Point Mutation ◽

Severe Bleeding ◽

Polymorphism Analysis ◽

Nucleotide Position ◽

Coagulation Factor Vii ◽

Fvii Deficiency

Abstract We have studied a family with homozygous lethal, blood coagulation factor VII (FVII) deficiency. To identify the mutation responsible for the deficiency, exons 2 to 8 and the intron-exon junctions of their FVII genes were amplified from peripheral white blood cell DNA by polymerase chain reaction and screened by single-strand conformational polymorphism analysis. The fragment showing aberrant mobility was cloned and sequenced. We detected a single point mutation, a homozygous G to A substitution at nucleotide position 6070, in the invariant GT dinucleotide at the 5′ splice site of intron 4. Homozygosity was confirmed by loss of a site for the restriction endonuclease Mlu I. Analysis of the splicing pattern of ectopic transcripts in lymphocytes in the parents revealed that this mutation is associated with skipping of exon 4, which produces an mRNA encoding FVII with an in-frame deletion of the first epidermal growth factor–like domain (EGF 1). Transient transfection of COS-7 cells with an expression vector containing the ▵EGF 1 FVII cDNA shows that this mutant protein is not expressed. The identification of the molecular basis of the FVII deficiency in this family allowed mutation-specific prenatal diagnosis to be performed in a subsequent pregnancy. In this family complete FVII deficiency is associated with a severe bleeding diathesis but no developmental abnormalities, lending weight to the hypothesis that fetal FVII is not required for the putative angiogenic functions of tissue factor in humans. © 1998 by The American Society of Hematology.

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Biochemical, molecular and clinical aspects of coagulation factor VII and its role in hemostasis and thrombosis

Haematologica ◽

10.3324/haematol.2020.248542 ◽

2021 ◽

Author(s):

Francesco Bernardi ◽

Guglielmo Mariani

Keyword(s):

Coagulation Factor ◽

Clinical Information ◽

Cardiovascular Death ◽

Clinical Findings ◽

Factor Vii ◽

Clinical Aspects ◽

Coagulation Factor Vii ◽

Wide Range ◽

Fvii Deficiency ◽

The Impact

Activated factor VII (FVIIa), the first protease of clotting, expresses its physiological procoagulant potential only after complexing with tissue factor (TF) exposed to blood. Deep knowledge of the FVIIa-TF complex and F7 gene helps to understand the Janus-faced clinical findings associated to low or elevated FVII activity (FVIIc). Congenital FVII deficiency, the most frequent among the recessively inherited bleeding disorders, is caused by heterogeneous mutations in the F7 gene. Complete FVII deficiency causes perinatal lethality. A wide range of bleeding symptoms, from life-threatening intracranial hemorrhage to mild mucosal bleeding, is observed in patients with apparently modest differences in FVIIc levels. Though clinically relevant FVIIc threshold levels are still uncertain, effective management, including prophylaxis, has been devised, substantially improving the quality of life of patients. The exposure of TF in diseased arteries fostered investigation on the role of FVII in cardiovascular disease. FVIIc levels were found to be predictors of cardiovascular death and to be markedly associated to F7 gene variation. These genotype-phenotype relationships are among the most extensively investigated in humans. Genome-wide analyses extended association to numerous loci that, together with F7, explain >50% of FVII level plasma variance. However, the ability of F7 variation to predict thrombosis was not consistently evidenced in the numerous population studies. Main aims of this review are to highlight i) the biological and clinical information that distinguishes FVII deficiency from the other clotting disorders and ii) the impact exerted by genetically predicted FVII level variation on bleeding as well as on the thrombotic states.

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An engineered human albumin enhances half-life and transmucosal delivery when fused to protein-based biologics

Science Translational Medicine ◽

10.1126/scitranslmed.abb0580 ◽

2020 ◽

Vol 12 (565) ◽

pp. eabb0580

Author(s):

Malin Bern ◽

Jeannette Nilsen ◽

Mattia Ferrarese ◽

Kine M. K. Sand ◽

Torleif T. Gjølberg ◽

...

Keyword(s):

Half Life ◽

Coagulation Factor ◽

Human Albumin ◽

Factor Vii ◽

Coagulation Factor Vii ◽

Activated Factor Vii ◽

Extended Plasma ◽

Albumin Variant ◽

Transcellular Delivery ◽

Plasma Half Life

Needle-free uptake across mucosal barriers is a preferred route for delivery of biologics, but the efficiency of unassisted transmucosal transport is poor. To make administration and therapy efficient and convenient, strategies for the delivery of biologics must enhance both transcellular delivery and plasma half-life. We found that human albumin was transcytosed efficiently across polarized human epithelial cells by a mechanism that depends on the neonatal Fc receptor (FcRn). FcRn also transported immunoglobulin G, but twofold less than albumin. We therefore designed a human albumin variant, E505Q/T527M/K573P (QMP), with improved FcRn binding, resulting in enhanced transcellular transport upon intranasal delivery and extended plasma half-life of albumin in transgenic mice expressing human FcRn. When QMP was fused to recombinant activated coagulation factor VII, the half-life of the fusion molecule increased 3.6-fold compared with the wild-type human albumin fusion, without compromising the therapeutic properties of activated factor VII. Our findings highlight QMP as a suitable carrier of protein-based biologics that may enhance plasma half-life and delivery across mucosal barriers.

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U1-snRNA–mediated rescue of mRNA processing in severe factor VII deficiency

Blood ◽

10.1182/blood-2007-10-117440 ◽

2008 ◽

Vol 111 (5) ◽

pp. 2681-2684 ◽

Cited By ~ 38

Author(s):

Mirko Pinotti ◽

Lara Rizzotto ◽

Dario Balestra ◽

Marzena Anna Lewandowska ◽

Nicola Cavallari ◽

...

Keyword(s):

Hepatoma Cell ◽

Coagulation Factor ◽

Mrna Processing ◽

Factor Vii ◽

Hepatoma Cell Line ◽

Exon Definition ◽

Coagulation Factor Vii ◽

U1 Snrna ◽

Fvii Deficiency ◽

Therapeutic Tools

Small nuclear U1-RNAs (snRNAs), the spliceosome components selectively recognizing donor splice sites (5′ss), were engineered to restore correct mRNA processing in a cellular model of severe coagulation factor VII (FVII) deficiency, caused by the IVS7 9726 + 5g/a change. Three U1-snRNAs, complementary to the mutated 5′ss (U1 + 5a) or to neighboring sequences were expressed with FVII minigenes in a hepatoma cell line. The U1-snRNAs reduced from 80% to 40% the exon 7 skipping, thus increasing exon definition. The U1 + 5a construct also dramatically increased recognition of the correct 5′ss over the 37-bp downstream cryptic site preferentially activated by the mutation, thus inducing appreciable synthesis of normal transcripts (from barely detectable to 50%). This effect, which was dose-dependent, clearly demonstrated that impaired recognition by the U1-snRNA was the mechanism responsible for FVII deficiency. These findings suggest compensatory U1-snRNAs as therapeutic tools in coagulation factor deficiencies caused by mutations at 5′ss, a frequent cause of severe defects.

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