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Reteplase - Formulation and Format (61)

Reteplase - Formulation and Format (61)

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Reteplase, a recombinant plasminogen activator


21 1


The purity of reteplase, determined by hgh resolution chromatographic

methods (Rp-HPLC and SE-HPLC) is as follows:

Reteplase protein makes up > 99.99% of the total protein.


E. coli protein

70 ppm

ETthrina trypsin irhbitor < 2 ppm

DNA< 25pg/lOU

Endotoxins< 5 E U / d



Reteplase is poorly soluble in water and the usual buffers, and requires the

addition of stabilizers (e.g. amino acids such as argtnine).


Absorption spectra (visible, UV, IR)

The protein concentration is determined by measurement of the absorption

at 280 nm against the respective buffer as reference. With a light path of 1 cm

and a protein concentration of 1 m g M , the absorbance is 1.69.


Isoelectric point (IEP)

The IEP of reteplase calculated from the amino acid composition is 7.23



The pH of the reconstituted medicinal product of reteplase is in the range

pH 7.0 to pH 7.4



If stored in the original package, the injection vials of reteplase 10 U are

stable for two years at temperatures of 2°C to 25°C. During prolonged

storage the powder should be protected from excessive exposure to light.

The lyophilisate is also stable if deep-frozen. Exposure to temperatures

above 25°C shortens the shelf-life; temperatures greater than 30°C should

be avoided.

After reconstitution as directed, the solution should be used immediately.

The reconstituted solution is chemically stable for 4 hours.



Dr Michael Waller and Dr Ulrich Kohnert

Nature and risk of the decomposition products

Decomposition results in transition to the two-chain form of reteplase.


Product presentation




Reteplase is presented as a sterile, dry, white, lyophilized powder in a

glass vial, mostly adhering to the glass wall of the vial.

Each vial contains 1.16 g powder, equivalent to 10 U (a single dose) of


Each dose of the medxinal product contains 1.O mg polysorbate 20, plus

871.0 mg arginine and 268.6 mg phosphoric acid as stabilizers.

The powder is reconstituted with 10 ml diluent (water) for intravenous



Potency units

The potency of reteplase is gven in units (U) based on a reference

standard which is reteplase-specific and is not comparable with the units used

for other thrombolytic agents. The designation Mu (1 MU = 1 U) is also

sometimes used in publications.


Ulrich Kohnert was the Project Leader Biotechnology for the development

of reteplase. He is a chemist by training and has special expertise in in vitro

folding, purification and in vitro characterization of proteins, as well as the

development of pharmaceutical formulations for therapeutic proteins.

Boehringer Mannheim is a htgh technology company in the healthcare market.

It holds a leading position in research development and production of

diagnostics and new drugs. Biotechnology has a prominent position and long

tradition withtn the company profile. Boehringer Mannheim operates one of

Europe's largest biotechnology facilities in Penzberg, Bavaria. Boehringer

Mannheim GmbH is part of the Roche Deutschland Holding GmbH.

Reteplase, a recombinant plasminogen activator



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Reteplme, a recombinant plasminogen activator


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Dr Michael Waller and Dr Ulrich Kohnert

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Boehringer Mannheim.

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