Natural killer stimulatory factor

• US 5,457,038 A
• Filed: 09/18/1990
• Issued: 10/10/1995
• Est. Priority Date: 11/10/1988
• Status: Expired due to Term

• Current Assignee: Genetics Institute LLC (Pfizer Inc.), Wyeth (Pfizer Inc.), The Wistar Institute
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Kobayashi, Michiko, Trinchieri, Giorgio, Perussia, Bice, Clark, Steven C., Wong, Gordon G., Hewick, Rodney
• Primary Examiner(s): Ellis, Joan
• Application Number:
  US07/584,941
• Time in Patent Office: 1,848 Days
• Field of Search: 435/69.1, 435/69.5, 435/91, 435/172.3, 435/235, 435/320.1, 435/240.2, 435/252.33, 435/69.52, 435/252.3, 435/255, 435/256, 536/27, 530/350, 935/18, 935/24, 935/37, 935/38, 935/41, 935/55, 935/61, 935/70, 935/72
• US Class Current: 435/69.52
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 14/5434 IL-12
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding

Abstract

A novel homogeneous human cytokine, Natural Killer Stimulatory Factor, having the ability to induce the production of gamma interferon in vitro in human peripheral blood lymphocytes, and a pharmaceutical preparation containing it.

Claims

1. A DNA sequence coding for NKSF, or a subunit thereof which possesses NKSF biological properties, which is selected from the group consisting of:

(a) the sequence of FIG. 1;

(b) a fragment of (a) which codes for a NKSF subunit having NKSF biological properties; and

(c) sequences capable of hybridizing thereto and which code for a protein having NKSF biological properties.

3. The DNA sequence of claim 1 having the sequence of FIG. 1.

8. A plasmid vector comprising a DNA sequence of claim 1.

9. The plasmid vector according to claim 8 wherein the vector is pNK40-4.

11. A cell transformed with the vector of claim 8 wherein the DNA sequence encoding NKSF is in operative association with an expression control sequence.

2. A DNA sequence coding for NKSF, or a subunit thereof which possesses NKSF biological properties, which is selected from the group consisting of:

(a) the sequence of FIG. 2;

(b) a fragment of (a) which codes for a NKSF subunit having NKSF biological properties; and

(c) sequences capable of hybridizing thereto and which code for a protein having NKSF biological properties.

4. The DNA sequence of claim 2 having the sequence of FIG. 2.

10. A plasmid vector comprising the DNA sequence according to claim 2 wherein the vector is p35nksf 14-1-1.

12. A cell transfected with (a)(i) a nucleic acid molecule encoding the 35 kD subunit of the protein identified as CLMF or NKSF, comprising the sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________ATG TGT CCA GCG CGC AGC CTC CTC CTT GTG GCT ACC CTG GTC CTC CTG GAC CAC CTC AGT TTG GCC AGA AAC CTC CCC GTG GCC ACT CCA GAC CCA GGA ATG TTC CCA TGC CTT CAC CAC TCC CAA AAC CTG CTG AGG GCC GTC AGC AAC ATG CTC CAG AAG GCC AGA CAA ACT CTA GAA TTT TAC CCT TGC ACT TCT GAA GAG ATT GAT CAT GAA GAT ATC ACA AAA GAT AAA ACC AGC ACA GTG GAG GCC TGT TTA CCA TTG GAA TTA ACC AAG AAT GAG AGT TGC CTA AAT TCC AGA GAG ACC TCT TTC ATA ACT AAT GGG AGT TGC CTG GCC TCC AGA AAG ACC TCT TTT ATG ATG GCC CTG TGC CTT AGT AGT ATT TAT GAA GAC TTG AAG ATG TAC CAG GTG GAG TTC AAG ACC ATG AAT GCA AAG CTT CTG ATG GAT CCT AAG AGG CAG ATC TTT CTA GAT CAA AAC ATG CTG GCA GTT ATT GAT GAG CTG ATG CAG GCC CTG AAT TTC AAC AGT GAG ACT GTG CCA CAA AAA TCC TCC CTT GAA GAA CCG GAT TTT TAT AAA ACT AAA ATC AAG CTC TGC ATA CTT CTT CAT GCT TTC AGA ATT CGG GCA GTG ACT ATT GAT AGA GTG ATG AGC TAT CTG AAT GCT TCC, and __________________________________________________________________________

(a) (ii) a second nucleic acid molecule encoding the 40 kD subunit of said CLMF or NKSF, comprising the

space="preserve" listing-type="tabular">__________________________________________________________________________ATG TGT CAC CAG CAG TTG GTC ATC TCT TGG TTT TCC CTG GTT TTT CTG GCA TCT CCC CTC GTG GCC ATA TGG GAA CTG AAG AAA GAT GTT TAT GTC GTA GAA TTG GAT TGG TAT CCG GAT GCC CCT GGA GAA ATG GTG GTC CTC ACC TGT GAC ACC CCT GAA GAA GAT GGT ATC ACC TGG ACC TTG GAC CAG AGC AGT GAG GTC TTA GGC TCT GGC AAA ACC CTG ACC ATC CAA GTC AAA GAG TTT GGA GAT GCT GGC CAG TAC ACC TGT CAC AAA GGA GGC GAG GTT CTA AGC CAT TCG CTC CTG CTG CTT CAC AAA AAG GAA GAT GGA ATT TGG TCC ACT GAT ATT TTA AAG GAC CAG AAA GAA CCC AAA AAT AAG ACC TTT CTA AGA TGC GAG GCC AAG AAT TAT TCT GGA CGT TTC ACC TGC TGG TGG CTG ACG ACA ATC AGT ACT GAT TTG ACA TTC AGT GTC AAA AGC AGC AGA GGC TCT TCT GAC CCC CAA GGG GTG ACG TGC GGA GCT GCT ACA CTC TCT GCA GAG AGA GTC AGA GGG GAC AAC AAG GAG TAT GAG TAC TCA GTG GAG TGC CAG GAG GAC AGT GCC TGC CCA GCT GCT GAG GAG AGT CTG CCC ATT GAG GTC ATG GTG GAT GCC GTT CAC AAG CTC AAG TAT GAA AAC TAC ACC AGC AGC TTC TTC ATC AGG GAC ATC ATC AAA CCT GAC CCA CCC AAG AAC TTG CAG CTG AAG CCA TTA AAG AAT TCT CGG CAG GTG GAG GTC AGC TGG GAG TAC CCT GAC ACC TGG AGT ACT CCA CAT TCC TAC TTC TCC CTG ACA TTC TGC GTT CAG GTC CAG GGC AAG AGC AAG AGA GAA AAG AAA GAT AGA GTC TTC ACG GAC AAG ACC TCA GCC ACG GTC ATC TGC CGC AAA AAT GCC AGC ATT AGC GTG CGG GCC CAG GAC CGC TAC TAT AGC TCA TCT TGG AGC GAA TGG GCA TCT GTG CCC TGC AGT;

or __________________________________________________________________________

(b) a nucleic acid molecule encoding both the 35 kD and 40 kD subunits of said CLMF or NKSF, comprising the sequences specified in (i) and (ii).

5. A process for producing NKSF, or a subunit thereof which possesses NKSF biological activity, comprising culturing a cell line transformed with a DNA sequence according to claim 12, wherein said DNA sequence is in operative association with an expression control sequence therefor.

6. The process according to claim 5 wherein said DNA sequence comprises the DNA sequence of FIG. 1.

7. The process according to claim 5 wherein said DNA sequence comprises the DNA sequence of FIG. 2.