Transferrin compositions to alleviate the side effects of...

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector

Reexamination Certificate

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C514S002600, C530S317000, C530S350000, C530S394000, C530S400000

Reexamination Certificate

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06328966

ABSTRACT:

BACKGROUND OF THE INVENTION
(i) Field of the Invention
The invention relates to the production of new drugs, particularly eutrophic drugs, the active principle of which consists of transferrins. The invention is of particular significance for human therapy. However it is not limited thereto. It may also be applicable in veterinary medicine.
(ii) Description of Related Art
An eutrophic drug means a drug that is capable of maintaining or restoring the structure and function of tissue and cells in the person's body, particularly when that person undergoes treatments with other drug principles which beyond the favourable clinical effects which they may exert, are also fraught with undesirable side effects, particularly are liable of seriously injuring healthy cells and tissue of the body. Thus no matter how valuable the drug, difficulties may be encountered in monitoring its use in patients heavily in need for it. Such type of drugs shall hereinafter be referred to as “cytotoxic drugs”.
An example of such cytotoxic drugs is cyclosporin which, as is well known, can also induce in the treated host muscular and functional disabilities accompanied by severe pains and/or nephrotoxicity that can ultimately produce renal dysfunction in the patient, for instance as evidenced by falls in glomerular filtration rates.
The dramatic side effects of antitumor drugs are well known. Similarly, anti-inflammatory drugs can entail a full array of damages ranging from gastric diseases to general disturbance of the metabolism in the treated patient.
There is thus a strong need to for a drug composition capable of overcoming the side effects of cytotoxic drugs, particularly to either prevent the tissue or cell degradation or promote the repair of damaged cells and tissue, more generally of assisting the natural principles which in the body participate to the eustasis.
The eutrophic drug of the composition of the invention is characterized in that its active principle consists of transferrins.
Although not always compulsory, the transferring should originate from the same mammal as the treated one. Thus human transferring should be the preferred active principle of eutrophic drugs for use in man.
Transferrins as such are a class of two-sited, single chain, metal-binding proteins, widely distributed in physiological fluids and cells of vertebrates.
Each transferrin consists of a single polypeptide chain, of molecular weight in the range 76000-81000, which contains two similar but not identical binding sites.
If the shape of human serum transferrin is approximated by an ellipsoid of revolution, then the ratio of major radii to minor is 2:1 for iron-saturated transferrin, and increases to 2.5:1 or 3:1 when the protein is freed from iron.
The isoelectric point of serum transferrin is on the acid side of neutrality.
The transferring are all glycoproteins.
Human serum transferrin contains about 5% carbohydrate, linked to the protein in two identical and nearly symmetrical branched heterosaccharide chains. it has a molecular weight of about 80000. 1 mg of the iron-saturated protein contains about 1.4 &mgr;g iron.
The complete amino acid sequence of human plasma transferrin has been established by at least three groups using CNBr cleavage (CNBr) and by complementary DNA (cDNA) methods (MacGillivray, RTA, et al. “The complete amino acid sequence of human serum transferrin”, Proc. Natl. Acad. Sci. USA 79, 2504-2508, 1982 and Uzan G. et al. “Molecular cloning and sequence analysis of cDNA for human transferrin” Biochem. Biophys. Res. Commun. 1990, 273-281, 1984 and Yang F. et al. “Human transferrin: cDNA characterization and chromosomal localization” Proc. Natl. Acad. Sci. USA 81, 2752-2756, 1984). It is composed of 678 amino acid residues, which together with the two-N-linked oligosaccharide chains exhibit a calculated molecular weight of 79,570 (of which 6% is contributed by the glucosidic moiety: MacGillivray, RTA et al. and Uzan G. et al., “Molecular cloning and sequence analysis of cDNA for human transferrin” Biochem. Biophys. Res. Commun. 119, 273-281, 1984). Williams J. (“The evolution of transferrin”, Trends Biochem. Sci. 7, 394-397, 1982) has suggested the importance of sulfhydryl groups in stabilizing the iron-binding site and has traced their evolutionary developement to the 17 disulfides found in human transferrin.
For a general review of the status of general knowledge about transferrins see the general publication titled “The physiology of transferrin and transferrin receptors” by Helmut A. Huebbers and Clement A. Finch in Phyiological Reviews, vol. 67, n° 2, Apr. 1987.
Procedures for obtaining transferrin, particularly transferrin of human origin in a biologically pure state have been disclosed in that publication. Preferred purification procedures are either based on physico-chemical based separation steps followed by selective fixation on matrix-bound antibody or matrix-bound receptor.
SUMMARY AND OBJECTS OF THE INVENTION
In a preferred embodiment of the invention, the transferrins used for the purpose of the invention consist of a mixture of transferrins obtained from a sufficient. number of donors. Advantageously, one uses “pools” of transferring, as they are obtainable from plasma pools produced in industry which can originate from several hundreds to several thousands of donors. As a matter of fact, it is believed that transferring obtained from different individual persons may have varying effects on the recipient, depending on their respective degrees of genetic similarities with said recipient. Advantageously, the transferrins for use in this invention result from the purification product obtained from a plasma resulting from the pooling of plasmas obtained from blood of at least 1000 donors.
The invention finds particularly advantageous uses in the protection of persons subjected to an immunosuppressive treatment, particularly with cyclosporin or other more or less related immunosuppressive drugs, e. g. FK506 and rapamycin. As a matter of fact, transferrins have been found to efficiently counteract the toxic effects of such drugs, as this will be further illustrated hereafter by the eustatic or eutrophic effect of transferrins on different metabolic functions in animals subject to treatments with cyclosporin, when they are either prolonged or carried out with high dosages.
As this will be seen hereafter, the results obtained at the histological level are spectacular. As well known the organs which are susceptible to the toxic effects of chronic administration of cyclosporin are particularly targetted on the thymus and even more so on the kidneys.
Transferrins are of particular interest in association with cyclosporin or other drugs having similar immunosuppressive effects, allowing for instance the graft of cells or tissue of a donor host in an allogeneic, or even xenogeneic recipient to succeed. As a matter of fact, transferrins are by themselves capable of inducing similar immunosuppressive effects under similar conditions in the recipient host, particularly for preventing graft-rejection, when administered together or concurrently with the grafting of the cells, tissue or organs in the recipient.
The invention however is not limited thereto.
The transferring can also be used to combat the toxic effects of other cytotoxic agents or compounds, particularly of those known to induce kidney damage to be substantially diminished, let alone eliminated. By way of example of such other drugs, one may mention:
antibiotics, particularly Polyene antibiotics, such as Colimycin and Gentamycin;
non-steroid anti-inflammatory drugs, particularly Ibuprofen;
anti-tumor drugs, such as Cis-platinum, or other immunosuppressive and cytotoxic drugs like Prednisolone or cyclophosphamide.
As already mentioned, the invention is not restricted to the use of transferrins in association with drugs which target principally the kidneys. Transferrins can also be used to prevent the damage of other tissues, for instance for the sake of preventing ear damages often produced by Gentamycin, Cis-platinum or streptomyc

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