New artificial oxygen carriers made of pegulated polymerised pyridoxylated porcine haemoglobin (P 4Hb)

Abstract

Oxygen-carrying plasma expanders are designed for use as iso-oncotic ‘blood substitutes’ to combat oxygen deficiencies caused by blood loss. In contrast, a hypo-oncotic artificial oxygen carrier can be added to existing blood — as a ‘blood additive’. It has potential therapeutic use for deficiencies of oxygen which are not entailed by blood (volume) lack, and can therefore not be treated by a ‘blood substitute’, e.g. anaemias, local ischaemias and their complications such as stroke or myocardial infarction, or lack of oxygen in tumours, reducing the effectiveness of anti-cancer treatments by irradiation or chemotherapy. For such a novel approach haemoglobin-based oxygen-carrying additive, the haemoglobin must be highly polymerised in order to decrease the oncotic pressure, which can be received many times lower compared with smaller molecular size haemoglobins. Our aim is to produce haemoglobin polymers with narrow distributions of molecular weights of approximately 1 000 000 g/mol, preferably produced in high yield and at low cost. But polymerising haemoglobin by cross-linking normally results in a so-called percolation distribution of molecular weights, with a large amount of insoluble material, and with only poor yields of the desired polymers. A newly developed one-vessel synthesis procedure, which includes a controlled marked dilution of the synthesis medium during the cross-linking reaction, enables yields of polymerised haemoglobin (P 4Hb) of over 80 %. Those preparations are easy and cheap to perform at large scales. P 4Hb hyperpolymers (the high molecular moiety of P 4Hb) are suitable for an oxygen-carrying blood additive: their oxygen-binding properties are sufficient, they are fully compatible with human blood plasma, and at the intended therapeutic concentration of approximately 30 g/l oncotic pressures are very low, and the impact on blood viscosity is tolerable.