Compositions containing glycosylated molecules having human tiss

Details Compositions containing glycosylated molecules having human tiss Compositions containing glycosylated molecules having human tiss

1991-07-19

1995-04-25

Robinson, Douglas W.

Drug, bio-affecting and body treating compositions

Enzyme or coenzyme containing

Hydrolases

424 941, 424 943, 435188, A61K 37547, A61K 3748, A61K 3762, C12N 996

Patent

active

054096995

DESCRIPTION:

BRIEF SUMMARY
Human tissue type plasminogen activator (t-PA) possesses great therapeutic importance in the dissolving of blood coagula, e.g. in the case of heart infarcts. t-PA brings about the dissolution of blood coagula by the activation of plasminogen to plasmin. Plasmin in turn dissolves fibrin, the main component of the protein matrix of coagulated blood.
Natural t-PA is composed of several functional domains F, E, K1, K2 and P. The domain P contains the proteolytically active region which brings about the cleavage of plasminogen to plasmin. Recombinant preparations of t-PA or of various t-PA muteins, in which one or more of the domains F, E, K1 and K2 are deleted, in eukaryotic and prokaryotic cells are already known. In contradistinction to natural t-PA, t-PA derivatives are synthesised from prokaryotes in non-glycosylated form.
Glycosylated proteins with t-PA activity only dissolve in low concentrations in the buffers usually employed for the solubilisation of proteins, such as e.g. 50 mmole/l. Na citrate, 50 mmole/l. phosphate or physiological NaCl solution. However, for their use as therapeutically-active material, protein solutions with a higher t-PA activity of at least 1.4 MU/ml., preferably of 1.4 MU/ml. to 10 MU/ml should be used.
It is known from EP-A 0 217 379 that neutral or slightly alkaline arginine formulations can increase the solubility of t-PA. A disadvantage of this process is, however, the low stability of highly concentrated solutions under neutral or slightly alkaline conditions.
U.S. Pat. No. 4,777,043 discloses a pharmaceutical composition with human t-PA and a pharmaceutically compatible argininium ion-containing buffer with a chloride ion concentration of up to 0.3 mole/l. EP-A 0 156 169, EP-A 0 303 351 and EP-A 0 297 294 disclose further possibilities of solubilising proteins with t-PA activity in buffers with particular amino acids, their salts, derivatives and homologues. Furthermore, t-PA can be stabilised by addition of gelatin according to EP-A 0 123 304, by addition of albumin according to EP-A 0 112 940 or by addition of a polysulphuric acid ester of a saccharide or of a sulphated sugar according to EP-A 0 198 321. PCT/US88/04402 discloses a process for increasing t-PA solubility, wherein one uses an aqueous medium with a basic amino acid, especially arginine, in a concentration of 0.02 to 0.2 mole/l., together with a citric acid group in a concentration of 0.02 to 0.08 mole/l. at a pH value of 5 to 8.
However, these various compositions are not generally suitable for all proteins with t-PA properties. Thus, it was ascertained that various glycosylated or non-glycosylated t-PA variants possess solubility properties which differ greatly from one another.
Consequently, it is the aim of the invention to develop pharmaceutical compositions which contain e.g., glycosylated t-PA or t-PA muteins with an activity of more than 1.4 MU/ml., whereby the t-PA is to be stable over a comparatively long period of time. The unit U is defined according to WHO, National Institute for Biological Standards and Control (cf. H. Lill, ZGIMAL, 42 (1987), 478-486).
According to the invention, the aim is solved by a pharmaceutical composition of a glycosylated protein with t-PA activity with an activity of at least 1.4 MU/ml. with a pH value of 4.5 to 9, whereby this composition contains citrate and at least one compound from the group consisting of C.sub.3 -C.sub.6 -cycloalkylene or benzylidene and R.sup.1 and R.sup.2, independently of one another, are H or C.sub.1 -C.sub.3 -alkyl, .delta.-aminovaleric acid, .gamma.-aminobutyric acid, further carboxyl group.
The concentration of the citrate ions in the pharmaceutical preparation according to the invention amounts to at least 5 mmole/l., preferably from 5 to 100 mmole/l. Especially preferred is a concentration of the citrate ions of 50 mmole/l. Depending upon the alkalinity of the compounds added, the pH value is adjusted with HCl or a base, such as e.g. NaOH or KOH.
It has proven to be suitable to adjust the pH value of the alkaline citrate solutions with

REFERENCES:
patent: 4927630 (1990-05-01), Feder et al.
patent: 4929444 (1990-05-01), Johnston et al.
patent: 4935237 (1990-06-01), Higgins et al.
patent: 4960702 (1990-10-01), Rig et al.
patent: 4980165 (1990-12-01), Isaacs et al.
patent: 4985245 (1991-01-01), Kikimoto et al.
patent: 5068106 (1991-11-01), Paques et al.
patent: 5100666 (1992-03-01), Bell et al.
patent: 5112609 (1992-05-01), Johnston et al.
patent: 5130143 (1992-07-01), Strickland et al.
patent: 5132214 (1992-07-01), Fedor et al.
patent: 5147643 (1992-10-01), Heyneker
patent: 5149533 (1992-10-01), Mulvihill et al.

Affiliated with

Also associated with

Rating

Compositions containing glycosylated molecules having human tiss does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Compositions containing glycosylated molecules having human tiss, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Compositions containing glycosylated molecules having human tiss will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-1566192