Recombinant clotting factors
Recombinant blood coagulation factors are genetically engineered analogues of coagulation factors occurring in human blood such as factor VII , factor VIII , factor IX , factor XIII or von Willebrand factor (VWF).
Recombinant coagulation factors are structurally very similar to the body's own coagulation factors and correspond to them in their mode of action. They are used therapeutically in deficiencies (such as hemophilia ) to prevent or treat bleeding. They are administered intravenously .
The production of recombinant coagulation factors is carried out with cells to which the human gene was used to express the corresponding coagulation factor. The cells are cultivated in a nutrient medium in bioreactors (fermentation tanks), where they form human protein and release it into the medium. From there it is extracted and purified. This biotechnological production method is not only more economical than obtaining the coagulation factors from human blood plasma , but also reduces the risk of contamination with pathogens such as HIV or HCV .
Development history
After the isolation and cloning of the factor VIII gene was successful in 1984, a recombinant factor VIII ( octocog alfa , rFVIII) was the first recombinant coagulation factor used therapeutically. It was approved for the treatment of haemophilia , first in 1992 in the USA and Sweden, and then in Germany in 1993 under the name Recombinate ( Baxter ) . Preparations from other companies followed, as well as other recombinant factors (rFVIIa, rFIX).
Newer generations of rFVIII preparations are characterized by the fact that they do not contain blood components ( albumin ) in the final formulation or even completely without the addition of human or animal plasma components in the entire manufacturing process (culture medium, cleaning), which further reduces the risk of biological contamination.
properties
The coagulation factors VII, VIII and IX are proteins that have sugar residues on their surface ( glycolised proteins, glycoproteins ). Factor VIII in particular is a very complex molecule: the protein part consists of 2332 amino acids, which are divided between a main and a side chain, and has 25 possible glycosylation sites. Since bacterial cells cannot glycolize recombinant proteins at all and yeast cells have difficulty glycolizing them, cell lines from mammalian cells are used. Kidney cells from baby hamsters ( BHK-21 cells) or ovarian cells from Chinese hamsters ( CHO cells ) are particularly suitable . The resulting glycolization profiles are very similar to those of the natural human coagulation factors.
By PEGylation , the leaves immunogenicity reduce the protease stability increase and renal slow excretion.
The short effectiveness of the rFIX can be extended by conjugation with serum proteins (fusion proteins).
As a non-glycosylated protein, factor XIII can be produced recombinantly with cells from baker's yeast .
Active ingredient overview
According to the INN nomenclature, the names of the recombinant coagulation factors end with -cog . Subordinate word stems are -eptacog , -octocog and -nonacog . The glycosylation pattern is characterized by the addition of a written Greek letter ( alfa , beta , etc.). Proteins with an amino acid sequence deviating from the natural model are identified by a prefix . Pegylated substances have the word pegol as a second addition . Activated shapes are marked with a corresponding bracket.
| Active ingredient, INN | Brief description | Cell line | Commercial preparation | Originator | Approval date |
|---|---|---|---|---|---|
| rFVIIa: -eptacog | |||||
| Eptacog alfa (activated) | Glycoprotein with 406 amino acids and a molecular mass of approx. 59 k Da . | BHK cells | NovoSeven | Novo Nordisk | 1996 (EU) |
| Eptacog alfa pegol (activated) | Pegylated eptacog alfa | (Experimental drug) | Novo Nordisk | - | |
| Vatreptacog alfa (activated) | Glycoprotein with 406 amino acids, modified amino acid sequence at positions 158, 296, 298. | (Experimental drug whose development has been discontinued) | Novo Nordisk | - | |
| Oreptacog alfa (activated) | Glycoprotein with 406 amino acids, modified amino acid sequence at positions 10, 32, 34, 36, 106, 253. | CHO cells | (Experimental drug) | Bayer | - |
| rFVIII: -octocog | |||||
| Octocog alfa | Glycoprotein with 2332 amino acids and a molecular mass of approx. 300 kDa. | CHO cells |
Recombinate Advate |
Baxter International | 1993 (D) 2004 (EU) |
| BHK cells |
Kogenate , Helixate NexGen Kovaltry |
Bayer | 2000 (EU) 2016 (EU) |
||
| Beroctocog alfa | Glycoprotein with 1424 amino acids, the B domain has been removed. | CHO cells | GreenGene | Green Cross Corporation | |
| Moroctocog alfa | Glycoprotein with 1438 amino acids and a molecular mass of 170 kDa. The B domain has been removed. | CHO cells | ReFacto A F | Pfizer | 1999 (EU) |
| Efmoroctocog alfa | Long-acting fusion protein with 1890 amino acids and a molecular mass of approx. 220 kDa. Consists of rFVIII, whose B domain has been removed and is covalently bound to the Fc fragment of human immunoglobulin G1 . | HEK cells | Elocta | Biogenic | 2015 (EU) |
| Damoctocog alfa pegol | Glycoprotein with 1438 amino acids and a molecular mass of 170 kDa. The B domain has been removed. Glycopegylated at Cys-1804. | BHK-21 cells | Jivi | Bayer | 2018 (EU) |
| Lonoctocog alfa | Glycoprotein with 1444 amino acids (single chain) with a molar mass of approx. 170 kDa. Most of the B domain and 4 amino acids of the adjacent acidic A3 domain are removed (amino acids 765 to 1652 of the full length of FVIII, including the furin cleavage site). | CHO cells | Afstyla | CSL Behring | 2017 (EU) |
| Rurioctocog alfa pegol | Glycoprotein, the 2332 amino acid protein of which bears a number of post-translational modifications, including more than 20 glycans , most of which are located on the B domain. The average molecular mass is about 330 kDa, of which the protein content makes up about 280 kDa. The PEG polymer has a mass of 20 kDa. | CHO cells | Adynovi | Baxalta | 2018 (EU) |
| Simoctocog alfa | Glycoprotein with 1440 amino acids and a molecular mass of approx. 170 kDa. Consisting of the factor VIII domains A1-A2 + A3-C1-C2, the B domain being replaced by a link sequence consisting of 16 amino acids. | HEK cells |
Nuwiq Vihuma (doublet) |
Octapharma | 2014 (EU) 2017 (EU) |
| Susoctocog alfa | Glycoprotein with 1448 amino acids and a molecular mass of approx. 175 kDa. Porcine sequence of factor VIII, the B domain is replaced by a link sequence consisting of 24 amino acids. | BHK-21 cells | Obizur | Baxalta | 2015 (EU) |
| Turoctocog alfa | Glycoprotein with 1445 amino acids and a molecular mass of about 166 kDA. The B-domain is shortened: it consists of 21 amino acids of the wild-type B-domain without any further modifications in the amino acid sequence. | CHO cells | NovoEight | Novo Nordisk | 2013 (EU) |
| Turoctocog alfa pegol | Turoctocog alfa glycopegylated on Ser-750. | CHO cells | Esperoct | Novo Nordisk | 2019 (USA) |
| rFIX: -nonacog | |||||
| Nonacog alfa | Single-chain glycoprotein with 415 amino acids and a relative molecular mass of approx. 55 kDa. The protein component corresponds to the allelic form Ala -148 of the plasmatic factor IX. | CHO cells | BeneFIX | Pfizer | 1997 (EU) |
| Nonacog beta pegol | Pegylated glycoprotein with 415 amino acids and a relative molecular mass of approx. 98 kDa (glycoprotein content: 56 kDa), protein component corresponds to the allelic form Ala-148 of plasmatic factor IX. | CHO cells | Refixia | Novo Nordisk | 2017 (EU) |
| Nonacog gamma | Single-chain glycoprotein with 415 amino acids and a relative molecular mass of approx. 68 kDa, protein component corresponds to the allelic form Ala-148 of plasmatic factor IX. | CHO cells | Rixubis | Baxter International | 2014 (EU) |
| Trenonacog alfa | Single-chain glycoprotein with 415 amino acids and a relative molecular mass of approx. 55 kDa, protein component corresponds to the Thr -148 variant of plasmatic factor IX. | CHO cells | (Experimental drug, 2013 application withdrawal in the EU) | Cangene Europe Ltd | - |
| Albutrepenonacog alfa | Long-acting fusion protein with 1018 amino acids; consisting of factor IX and human albumin. The amino acid sequence of rF IX corresponds to the Thr-148 variant of the plasmatic factor IX. | CHO cells | Idelvion | CSL Behring | 2016 (EU) |
| Eftrenonacog alfa | Long-acting fusion protein with 867 amino acids and a molecular mass of approx. 98 kDa; consists of factor IX, which is covalently bound to the Fc fragment of human immunoglobulin G1. The amino acid sequence of rF IX corresponds to the Thr-148 variant of the plasmatic factor IX. | HEK cells | Alprolix | Biogen Idec | 2016 (EU) |
| rFXIII: -tridecacog | |||||
| Catridecacog | Dimer made up of two factor XIII A subunits with a molecular mass of approx. 166 kDa, not glycosylated. | Yeast cells | NovoThirteen | Novo Nordisk | 2012 (EU) |
| rVWF: -vonicog | |||||
| Vonicog alfa | Non-glycated protein made up of 2050 amino acids. [Thr-618, Asp -709] Von Willebrand factor Homo sapiens (1381A> T, 1472H> D variant) |
CHO cells | Vonvendi / Veyvondi | Shire PLC | 2018 (EU) |
application areas
Eptacog alfa (activated) is indicated for the treatment of bleeding and the prevention of bleeding associated with surgical or invasive procedures in patients with certain types of congenital or acquired haemophilia. The recombinant factors VIII and IX are used for the treatment and prophylaxis of bleeding in haemophilia A and B, the recombinant factor XIII for the long-term prophylaxis of bleeding in a congenital deficiency of factor XIII-A subunits.
A recombinantly produced Von Willebrand factor has been available since 2018 (Vonvendi / Veyvondi from Shire, which was previously developed as "BAX 111" by Baxter). These factor concentrates could be used in the future to treat Willebrand-Jürgens syndrome and replace preparations such as haemates made from human plasma .
Side effects
The most common adverse effects include hypersensitivity reactions and allergic reactions. In addition, inhibitors can form, so that the anticoagulant effect is weakened. In such cases, therapy with the bispecific monoclonal antibody emicizumab (approved so far in the USA, in the evaluation in the EU) can be an alternative, as it is also effective in the formation of inhibitors.
Web links
- Theo Dingermann: Recombinant Active Ingredients. Special features of a special class of active ingredients . In: Österreichische Apothekerzeitung , issue 25 of December 9, 2008
- Therapy with coagulation factors , German Haemophilia Society for Combating Bleeding Diseases eV
Individual evidence
- ↑ International Nonproprietary Names (INN) for biological and biotechnological substances (a review) , World Health Organization (WHO), 2013.
- ↑ Vonicog alfa (Veyvondi) • DRUG-NEWS. Retrieved October 3, 2019 .