EP0158286A2
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EP
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A novel human physiologically active polypeptide
Assignee: Asahi Kasei Corp
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Filing: 1985-04-04
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Granted: —
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Expires: 2005-04-04
Abstract
Abstract A human physiologically active polypeptide, human Tumor Necrosis Factor (human TNF), comprising a specific amino acid sequence of 155 amino acid residues. The base sequence of the DNA coding for the human TNF has been determined using rabbit TNF cDNA. The human TNF can be advantageously produced on a large scale by recombinant DNA technique. The human TNF of the present invention has been found to be excellent in inducing necrosis of tumors with no toxic effect upon the normal tissues of the living body.
Inventors
- 👤 Robert Bruce Wallace
- 👤 Hirataka Itoh
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 23:52
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/525; A; A61; A61P; A61P31/00; A61P31/12; A61P35/00; C12P; C12P21/00; C12P21/02; A61K; A61K38/00; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948; Y10S530/00; Y10S530/827; Y10S530/828
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
102 words
A human physiologically active polypeptide having an amino acid sequence represented by the following formula (I): wherein Gln stands for a glutamine residue, Asp an aspartic acid residue, Pro a proline residue, Tyr a tyrosine residue, Val a valine residue, Lys a lysine residue, Glu a glutamic acid residue, Ala an alanine residue, Asn an asparagine residue, Leu a leucine residue, Phe a phenylalanine residue, Gly a glycine residue, His a histidine residue, Ser a serine residue, Thr a threonine residue, Ile an isoleucine residue, Trp a tryptophan residue, Arg an arginine residue, Met a methionine residue, and Cys a cysteine residue.
Claim 2
Dependent Claim
102 words
A human physiologically active polypeptide having an amino acid sequence represented by the following formula (I): wherein Gln stands for a glutamine residue, Asp an aspartic acid residue, Pro a proline residue, Tyr a tyrosine residue, Val a valine residue, Lys a lysine residue, Glu a glutamic acid residue, Ala an alanine residue, Asn an asparagine residue, Leu a leucine residue, Phe a phenylalanine residue, Gly a glycine residue, His a histidine residue, Ser a serine residue, Thr a threonine residue, Ile an isoleucine residue, Trp a tryptophan residue, Arg an arginine residue, Met a methionine residue, and Cys a cysteine residue.
Claim 3
Dependent Claim
116 words
A deoxyribonucleic acid comprising a base sequence coding for a human physiologically active polypeptide, said human physiologically active polypeptide having an amino acid sequence represented by the following formula (I): wherein Gln stands for a glutamine residue, Asp an aspartic acid residue, Pro a proline residue, Tyr a tyrosine residue, Val a valine residue, Lys a lysine residue, Glu a glutamic acid residue, Ala an alanine residue, Asn an asparagine residue, Leu a leucine residue, Phe a phenylalanine residue, Gly a glycine redidue, His a histidine residue, Ser a serine residue, Thr a threonine residue, Ile an isoleucine residue, Trp a tryptophan residue, Arg an arginine residue, Met a methionine residue, and Cys a cysteine residue.
Claim 4
Dependent Claim
116 words
A deoxyribonucleic acid comprising a base sequence coding for a human physiologically active polypeptide, said human physiologically active polypeptide having an amino acid sequence represented by the following formula (I): wherein Gln stands for a glutamine residue, Asp an aspartic acid residue, Pro a proline residue, Tyr a tyrosine residue, Val a valine residue, Lys a lysine residue, Glu a glutamic acid residue, Ala an alanine residue, Asn an asparagine residue, Leu a leucine residue, Phe a phenylalanine residue, Gly a glycine redidue, His a histidine residue, Ser a serine residue, Thr a threonine residue, Ile an isoleucine residue, Trp a tryptophan residue, Arg an arginine residue, Met a methionine residue, and Cys a cysteine residue.
Claim 5
Dependent Claim
79 words
3 . A deoxyribonucleic acid comprising at least one base sequence selected from the group consisting of a base sequence represented by the following formula (II) and a complementary base sequence to said base sequence: wherein A stands for a deoxyadenylic acid residue, G a deoxyguanylic acid residue, C a deoxycytidylic acid residue and T thymidylic acid residue and wherein the left end and right end of the formula (II) represent 5'-hydroxyl group side and 3'-hydroxyl group side, respectively.
Claim 6
Dependent Claim
79 words
3 . A deoxyribonucleic acid comprising at least one base sequence selected from the group consisting of a base sequence represented by the following formula (II) and a complementary base sequence to said base sequence: wherein A stands for a deoxyadenylic acid residue, G a deoxyguanylic acid residue, C a deoxycytidylic acid residue and T thymidylic acid residue and wherein the left end and right end of the formula (II) represent 5'-hydroxyl group side and 3'-hydroxyl group side, respectively.
Claim 7
Dependent Claim
38 words
A deoxyriponucleic acid comprising a base sequence which is one obtained by substituting at least one base of said at least one base sequence of the deoxyribonucleic acid of claim 3 in accordance with degeneracy of genetic code.
Claim 8
Dependent Claim
19 words
_5. A replicable recombinant DNA which comprises -a deoxyribonucleic acid according to claim 2 and a replicable expression vehicle.
Claim 9
Dependent Claim
15 words
A microorganism or cell culture transformed with a replicable recombinant DNA according to claim 5.
Claim 10
Independent Claim
212 words
A method for producing a human physiologically active polypeptide having TNF activity which comprises: (a) ligating to a replicable expression vehicle a deoxyribonucleic acid comprising a base sequence coding for a human physiologically active polypeptide, said human physiologically active polypeptide having an amino acid sequence represented by the following formula (I): wherein Gln stands for a glutamine residue, Asp an aspartic acid residue, Pro a proline residue, Tyr a tyrosine residue, Val a valine residue, Lys a lysine residue, Glu a glutamic acid residue, Ala an alanine residue, Asn an asparagine residue, Leu a leucine residue, Phe a phenylalanine residue, Gly a glycine residue, His a histidine residue, Ser a serine residue, Thr a threonine residue, Ile an isoleucine residue, Trp a tryptophan residue, Arg an arginine residue, Met a methionine residue, and Cys a cysteine residue. to obtain-a replicable recombinat DNA comprising said deoxyribonucleic acid and said replicable expression vehicle; (b) transforming cells of a microorganism or cell culture with said replicable recombinat DNA to form transformants; (c) selecting said transformants from parent cells of the microorganism or cell culture; (d) incubating said transformants, causing said transformants to express said deoxyribonucleic acid and produce a human physiologically active polypeptide; and (e) isolating said human physiologically active polypeptide from the incubated transformants.
Claim 11
Independent Claim
212 words
A method for producing a human physiologically active polypeptide having TNF activity which comprises: (a) ligating to a replicable expression vehicle a deoxyribonucleic acid comprising a base sequence coding for a human physiologically active polypeptide, said human physiologically active polypeptide having an amino acid sequence represented by the following formula (I): wherein Gln stands for a glutamine residue, Asp an aspartic acid residue, Pro a proline residue, Tyr a tyrosine residue, Val a valine residue, Lys a lysine residue, Glu a glutamic acid residue, Ala an alanine residue, Asn an asparagine residue, Leu a leucine residue, Phe a phenylalanine residue, Gly a glycine residue, His a histidine residue, Ser a serine residue, Thr a threonine residue, Ile an isoleucine residue, Trp a tryptophan residue, Arg an arginine residue, Met a methionine residue, and Cys a cysteine residue. to obtain-a replicable recombinat DNA comprising said deoxyribonucleic acid and said replicable expression vehicle; (b) transforming cells of a microorganism or cell culture with said replicable recombinat DNA to form transformants; (c) selecting said transformants from parent cells of the microorganism or cell culture; (d) incubating said transformants, causing said transformants to express said deoxyribonucleic acid and produce a human physiologically active polypeptide; and (e) isolating said human physiologically active polypeptide from the incubated transformants.
Claim 12
Dependent Claim
15 words
A substantially pure physiologically active polypeptide having the amino acid sequence defined in claim 1.
Claim 13
Dependent Claim
10 words
A substantially pure physiologically active polypeptide having human TNF activity.
Claim 14
Dependent Claim
10 words
A substantially pure physiologically active polypeptide having human TNF activity.
Claim 15
Dependent Claim
18 words
A substantially pure deoxyribonucleic acid comprising a base sequence coding for a polypeptide having human T NF activity.
Claim 16
Dependent Claim
18 words
A substantially pure deoxyribonucleic acid comprising a base sequence coding for a polypeptide having human T NF activity.
Claim 17
Dependent Claim
12 words
A substantially pure deoxyribonucleic acid comprising the sequence defined in claim 3.
Claim 18
Dependent Claim
18 words
A plasmid or bacteriophage transfer vector comprising a base sequence coding for a polypeptide having human TNF activity.
Claim 19
Dependent Claim
18 words
A plasmid or bacteriophage transfer vector comprising a base sequence coding for a polypeptide having human TNF activity.
Claim 20
Dependent Claim
18 words
A plasmid or bacteriophage transfer vector comprising a base sequence coding for a polypeptide defined in claim 1.
Claim 21
Dependent Claim
18 words
A microorganism or cell culture transformed by the transfer vector of claim 12, and mutants or variants thereof.
Claim 22
Dependent Claim
18 words
A microorganism or cell culture transformed by the transfer vector of claim 13, and mutants or variants thereof.
Claim 23
Dependent Claim
8 words
The microorganism E. coli k-12 strain JM83 (pRGE).
Claim 24
Dependent Claim
8 words
The microorganism E. coli k-12 strain JM83 (pRGE).
Claim 25
Dependent Claim
8 words
The microorganism E. coli k-12 strain JM83 (pHGE).
Claim 26
Dependent Claim
8 words
The microorganism E. coli k-12 strain JM83 (pHGE).
Claim 27
Dependent Claim
3 words
The plasmid pRGE.
Claim 28
Dependent Claim
3 words
The plasmid pRGE.
Claim 29
Dependent Claim
3 words
The plasmid pHGE.
Claim 30
Dependent Claim
3 words
The plasmid pHGE.
Claim 31
Dependent Claim
28 words
A pharmaceutical composition comprising an effective antitumor or anti-viral amount of a polypeptide as defined in claim 9 and at least one pharmaceutically acceptable carrier, diluent or excipient.
Claim 32
Dependent Claim
28 words
A pharmaceutical composition comprising an effective antitumor or anti-viral amount of a polypeptide as defined in claim 1 and at least one pharmaceutically acceptable carrier, diluent or excipient.
References Cited (Backward Citations)
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