Highly purified human growth hormone was isolated from fresh glands by the
procedure involving gel filtration of Sephadex. The hormone was oxidized by
performing acid. Both the native and the oxidized product were submitted to
digestion with trypsin as well as with chymotrypsin, while only native hormone
was hydrolyzed with pepsin. These digests were fractionated on ion-exchange
resin columns. Some of fraction required further purification by paper
chromatography in a system consisting of 1-butanol/acetic acid/water (4:1:5)
and by high-voltage electrophoresis on paper in a buffer of pH 2.3. Human
growth hormone is contained 188 amino acids. From sedimentation equilibre studies the molecular weight of the hormone was shown to be 21,500.
1000 ml of sterile intramuscular preparation containing 2.5 mg of
chymotripsin hydrolyzed bovine growth hormone and 2.5 mg
methylprednisolone is prepared from the following types and amounts of
materials containing (per ml); 1) 2.5 mg chymotripsin hydrolyzed bovine
growth hormone (powder), 2) 2.5 mg methylprednisolone (micronized), 3) 30
mg polyethylene glycol (40 000), 4) 9 mg sodium chloride, 5) 2 mg
polysorbate 80, 1.8 mg methylparaben, 0.2 mg propylparaben and watef q.s.
to 1000 ml.
The parabens are added to most of the water and dissolved with stirring and
heating to 65-70°C. The mixture is cooled to room temperature and
ingredient 3, 4 and 5 are added. The resulting solution was made up to 100
ml with water and sterilized by filtration. Ingredient 1 and 2 are sterilized with
ethylene oxide and added aseptically. The final preparation is packaged in
sterile containers with 1 ml of the final product. This is used advantageously
in the treatment of arthritis and to avoid negative nitrogene balance.
