Method of ameliorating the harmful effects of TNF using a...

Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai

Reexamination Certificate

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C514S008100, C514S012200, C514S021800, C424S085100, C424S085200

Reexamination Certificate

active

06417158

ABSTRACT:

FIELD OF THE INVENTION
The invention is in the field of recombinant genetics. In particular, the invention relates to a TNF receptor and to a TNF binding protein produced by recombinant means.
BACKGROUND OF THE INVENTION
Tumour necrosis factor (TNF-&agr;) was first found in the serum of mice and rabbits which had been infected with Bacillus Calmette-Guerin and which had been injected with endotoxin, and was recognized on the basis of its cytotoxic and antitumor properties (Carswell, E.A., et al.,
Proc. Natl. Acad. Sci.
25: 3666-3670 (1975)). It is produced particularly by activated macrophages and monocytes.
Numerous types of cells which are targets of TNF have surface receptors with a high affinity for this polypeptide (Old, L. J.,
Nature
326:330-331 (1987)); it was assumed that lymphotoxin (TNF-P) binds to the same receptor (Aggarwal, B. B., et al.,
Nature
318:655-667 (1985); Gullberg, U., et al.,
Eur. J. Haematol.
39:241-251 (1987)). TNF-&agr; is identical to a factor referred to as cachectin (Beutler, B., et al.,
Nature
316:552-554 (1985)) which suppresses lipoprotein lipase and results in hypertriglyceridaemia in chronically inflammatory and malignant diseases (Torti, F. M. et al.,
Nature
229:867-869 (1985); Mahoney, J. R., et al.,
J. Immunol.
134:1673-1675 (1985)). TNF-&agr; would appear to be involved in growth regulation and in the differentiation and function of cells which are involved in inflammation, immune processes and hematopoieses.
TNF can have a positive effect on the host organism by stimulating neutrophils (Shalaby, M. R., et al.,
J. Immunol.
135:2069-2073 (1985); Klebanoff, S. J., et al.,
J. Immunol.
136:4220-4225 (1986)) and monocytes and by inhibiting the replication of viruses (Mestan, J., et al.,
Nature
323:816-819 (1986); Wong, G. H. W., et al.,
Nature
323:819-822 (1986)). Moreover, TNF-&agr; activates the immune defenses against parasites and acts directly and/or indirectly as a mediator in immune reactions, inflammatory processes and other processes in the body, although the mechanisms by which it works have not yet been clarified in a number of cases. However, the administration of TNF-&agr; (Cerami, A., et al.,
Immunol. Today
9:28-31 (1988)) can also be accompanied by harmful phenomena (Tracey, K. J., et al.,
Science
234:470-474 (1986)) such as shock and tissue damage, which can be remedied by means of antibodies against TNF-&agr; (Tracey, K. J., et al.,
Nature
330:662-666 (1987)).
A number of observations lead one to conclude that endogenously released TNF-&agr; is involved in various pathological conditions. Thus, TNF-&agr; appears to be a mediator of cachexia which can occur in chronically invasive, e.g. parasitic, diseases. TNF-&agr; also appears to play a major part in the pathogenesis of shock caused by gram negative bacteria (endotoxic shock); it would also appear to be implicated in some if not all the effects of lipopolysaccharides (Beutler B., et al.,
Ann. Rev. Biochem.
57:505-18 (1988)). TNF has also been postulated to have a function in the tissue damage which occurs in inflammatory processes in the joints and other tissues, and in the lethality and morbidity of the graft-versus-host reaction (GVHR, Transplant Rejection (Piguet, P. F., et al.,
Immunobiol.
175:27 (1987)). A correlation has also been reported between the concentration of TNF in the serum and the fatal outcome of meningococcal diseases (Waage, A., et al.,
Lancet ii:
355-357 (1987)).
It has also been observed that the administration of TNF-&agr; over a lengthy period causes a state of anorexia and malnutrition which has symptoms similar to those of cachexia, which accompany neoplastic and chronic infectious diseases (Oliff A., et al.,
Cell
555-63 (1987)).
It has been reported that a protein derived from the urine of fever patients has a TNF inhibiting activity; the effect of this protein is presumed to be due to a competitive mechanism at the level of the receptors (similar to the effect of the interleukin-1 inhibitor (Seckinger, P., et al.,
J. Immunol.
139:1546-1549 (1987); Seckinger P., et al.,
J. Exp. Med.,
1511-16 (1988)).
EP-A2 308 378 describes a TNF inhibiting protein obtained from human urine. Its activity was demonstrated in the urine of healthy and ill subjects and determined on the basis of its ability to inhibit the binding of TNF-&agr; to its receptors on human HeLa cells and FS 11 fibroblasts and the cytotoxic effect of TNF-&agr; on murine A9 cells. The protein was purified until it became substantially homogeneous and characterized by its N-ending. This patent publication does indeed outline some theoretically possible methods of obtaining the DNA coding for the protein and the recombinant protein itself; however, there is no concrete information as to which of the theoretically possible solutions is successful.
SUMMARY OF THE INVENTION
The invention relates to DNA coding for a TNF receptor protein or a fragment thereof. In particular, the invention relates to DNA coding for the TNF receptor protein having the formula
ATG GGC CTC TCC ACC GTG CCT GAC CTG CTG CTG CCA CTG GTG

CTC CTG GAG CTG TTG GTG GGA ATA TAC CCC TCA GGG GTT ATT

GGA CTG GTC CCT CAC CTA GGG GAC AGG GAG AAG AGA GAT AGT

GTG TGT CCC CAA GGA AAA TAT ATC CAC CCT CAA AAT AAT TCG

ATT TGC TGT ACC AAG TGC CAC AAA GGA ACC TAC TTG TAC AAT

GAC TGT CCA GGC CCG GGG CAG GAT ACG GAC TGC AGG GAG TGT

GAG AGC GGC TCC TTC ACC GCT TCA GAA AAC CAC CTC AGA CAC

TGC CTC AGC TGC TCC AAA TGC CGA AAG GAA ATG GGT CAG GTG

GAG ATC TCT TCT TGC ACA GTG GAC CGG GAC ACC GTG TGT GGC

TGC AGG AAG AAC CAG TAC CGG CAT TAT TGG AGT GAA AAC CTT

TTC CAG TGC TTC AAT TGC AGC CTC TGC CTC AAT GGG ACC GTG

CAC CTC TCC TGC CAG GAG AAA CAG AAC ACC GTG TGC ACC TGC

CAT GCA GGT TTC TTT CTA AGA GAA AAC GAG TGT GTC TCC TGT

AGT AAC TGT AAG AAA AGC CTG GAG TGC ACG AAG TTG TGC CTA

CCC CAG ATT GAG AAT GTT AAG GGC ACT GAG GAC TCA GGC ACC

ACA GTG CTG TTG CCC CTG GTC ATT TTC TTT GGT CTT TGC CTT

TTA TCC CTC CTC TTC ATT GGT TTA ATG TAT CGC TAC CAA CGG

TGG AAG TCC AAG CTC TAC TCC ATT GTT TGT GGG AAA TCG ACA

CCT GAA AAA GAG GGG GAG CTT GAA GGA ACT ACT ACT AAG CCC

CTG GCC CCA AAC CCA AGC TTC AGT CCC ACT CCA GGC TTC ACC

CCC ACC CTG GGC TTC AGT CCC GTG CCC AGT TCC ACC TTC ACC

TCC AGC TCC ACC TAT ACC CCC GGT GAC TGT CCC AAC TTT GCG

GCT CCC CGC AGA GAG GTG GCA CCA CCC TAT CAG GGG GCT GAC

CCC ATC CTT GCG ACA GCC CTC GCC TCC GAC CCC ATC CCC AAC

CCC CTT CAG AAG TGG GAG GAC AGC GCC CAC AAG CCA C

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