EP0309050A1
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Human insulin-like growth factor analogs with reduced binding to serum carrier proteins and their production in yeast
Assignee: Merck and Co Inc
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Filing: 1988-09-16
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Granted: —
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Expires: 2008-09-16
Abstract
Abstract A synthetic gene encoding a 71-amino acid analog of human insulin-like growth factor (hIGF-I) has been constructed and expressed in the yeast, Saccharomyces cerevisiae . The protein analog, IGF132, contains the first 17 amino acids of the B chain of human insulin in place of the first 16 amino acids of hIGF-I. The purified hybrid protein has high affinity for the type I IGF receptor (12 nM) yet has drastically reduced affinity for human serum carrier proteins (>1000 nM). This analog is 5 to 10 times more active than normal hIGF-I in stimulating DNA synthesis in 3T3 cells and is a more active growth factor in vivo due to its reduced affinity for serum carrier proteins. Other proteins with similar properties have also been constructed. The protein analogs thus have a variety of utilities such as in promoting lactation in animals; promoting growth and feed efficiency in animals; improving carcass quality by increasing lean and decreasing fat; promoting wound healing in animals, including humans; promoting glucose utilization in skeletal muscle, and stimulating erythropoiesis, the production of red blood cells.
Inventors
- 👤 Joy D. Applebaum
- 👤 Marvin L. Bayne
- 👤 Margaret A. Cascieri
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-07 13:41
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/65; A; A61; A61P; A61P15/00; A61P43/00; A61P7/00; A61P7/06; C07K14/62; A61K; A61K38/00; C07K2319/00
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
159 words
- A process for the preparation of the synthetic polypeptide analog of hIGF-I by the recombinant DNA expression systems of bacteria, yeast or tissue culture cells hosts, said polypeptide analog having the following structure: A₁-A₂-A₃-A₄-LCG-A₅-A₆-LV-A₇-AL-A₈-A₉-R wherein: A₁ is G, V, or FV; A₂ is P or N; A₃ is E or Q; A₄ is T, H or A; A₅ is A or S; A₆ is E or H; A₇ is D or E; A₈ is Q or Y; A₉ is F or L; and R is the remainder of the hIGF-I peptide, provided that and A₁ to A₉ groups and the other amino acids do not constitute GPETLCGAELVDALQF-R, which comprises: a) insertion of the appropriate synthetic gene into an expression vector to form an expression cassette; b) introduction of the expression cassette into the bacteria, yeast or tissue cell culture host; c) growth of the transformed expression host; and d) purification of the desired polypeptide analog from said host.
Claim 2
Dependent Claim
159 words
- A process for the preparation of the synthetic polypeptide analog of hIGF-I by the recombinant DNA expression systems of bacteria, yeast or tissue culture cells hosts, said polypeptide analog having the following structure: A₁-A₂-A₃-A₄-LCG-A₅-A₆-LV-A₇-AL-A₈-A₉-R wherein: A₁ is G, V, or FV; A₂ is P or N; A₃ is E or Q; A₄ is T, H or A; A₅ is A or S; A₆ is E or H; A₇ is D or E; A₈ is Q or Y; A₉ is F or L; and R is the remainder of the hIGF-I peptide, provided that and A₁ to A₉ groups and the other amino acids do not constitute GPETLCGAELVDALQF-R, which comprises: a) insertion of the appropriate synthetic gene into an expression vector to form an expression cassette; b) introduction of the expression cassette into the bacteria, yeast or tissue cell culture host; c) growth of the transformed expression host; and d) purification of the desired polypeptide analog from said host.
Claim 3
Dependent Claim
46 words
- The process of claim 1, wherein A₁ is G, V, or FV; A₂ is P or N; A₃ is Q; A₄ is A; A₅ is A or S; A₆ is E or H; A₇ is D or E; A₈ is Y; and A₉ is L.
Claim 4
Dependent Claim
12 words
- The process of claim 1, wherein the polypeptide obtained is FVNQHLCGSHLVEALYL-R.
Claim 5
Dependent Claim
12 words
- The process of claim 1, wherein the polypeptide obtained is GPETLCGAELVDALYL-R.
Claim 6
Dependent Claim
12 words
- The process of claim 1, wherein the polypeptide obtained is GPQALCGAELVDALOF-R.
Claim 7
Dependent Claim
12 words
- The process of claim 1, wherein the polypeptide obtained is GPQALCGAELVDALYL-R.
Claim 8
Dependent Claim
11 words
- The process of claim 1, wherein the polypeptide obtain-is VNQHLCGSHLVEALYL-R.
Claim 9
Dependent Claim
48 words
- A process for the preparation of a synthetic gene encoding for the polypeptide obtained in claim 1, which comprises: a) the synthesis of the appropriate constituent oligonucleotides; b) annealing and ligation of said oligonucleotides to gene fragments; and c) cloning of synthetic gene into recombinant DNA plasmid.
Claim 10
Dependent Claim
22 words
- A process according to claim 8 for the preparation of the synthetic gene encoding for the polypeptide obtained in claim 3.
Claim 11
Dependent Claim
153 words
- The process of claim 9, wherein the synthetic gene obtained is: CAT TTG TGT GGC TCC CAT CTC GTT GAA GCT TTG TAC TTG GTA AAC ACA CCG AGG GTA GAG CAA CTT CGA AAC ATG AAC GTT TGC GGT GAC CGC GGT TTC TAC TTC AAC AAA CCG ACT CAA ACG CCA CTG GCG CCA AAG ATG AAG TTG TTT GGC TGA GGT TAC GGT TCT TCT TCT AGA CGT GCT CCG CAG ACT GGT CCA ATG CCA AGA AGA AGA TCT GCA CGA GGC GTC TGA CCA ATC GTT GAT GAA TGC TGC TTC AGA TCT TGT GAC CTG CGT TAG CAA CTA CTT ACG ACG AAG TCT AGA ACA CTG GAC GCA CGT CTC GAG ATG TAC TGC GCA CCG CTG AAA CCG GCT AAA GCA GAG CTC TAC ATG ACG CGT GGC GAC TTT GGC CGA TTT TCT GCT TGA TAA GTCG AGA CGA ACT ATT CAGCC TAG
Claim 12
Dependent Claim
22 words
- A process according to claim 8 for the preparation of the synthetic gene encoding for the polypeptide obtained in claim 4.
Claim 13
Dependent Claim
172 words
- The process of claim 11, wherein the synthetic gene obtained is: ATC CTT TCC TTG GAT AAA AGA GGT CCG GAA ACT TTG TGT TAG GAA AGG AAC CTA TTT TCT CCA GGC CTT TGA AAC ACA GGT GCT GAG CTC GTT GAC GCT CTG TAC CTC GTT TGC CCA CGA CTC GAG CAA CTG CGA GAC ATG GAG CAA ACG GGT GAC CGC GGT TTC TAC TTC AAC AAA CCG ACT GGT TAC CCA CTG GCG CCA AAG ATG AAG TTG TTT GGC TGA CCA ATG GGT TCT TCT TCT AGA CGT GCT CCG CAG ACT GGT ATC GTT CCA AGA AGA AGA TCT GCA CGA GGC GTC TGA CCA TAG CAA GAT GAA TGC TGC TTC AGA TCT TGT GAC CTG CGT CGT CTA CTT ACG ACG AAG TCT AGA AGA CTG GAC GCA GCA CTC GAG ATG TAC TGC GCA CCG CTG AAA CCG GCT AAA TCT GAG CTC TAC ATG ACG CGT GGC GAC TTT GGC CGA TTT AGA GCT TGA TAA GTCG CGA ACT ATT CAGCCTAG
Claim 14
Dependent Claim
22 words
- A process according to claim 8, for the preparation of the synthetic gene encoding for the polypeptide obtained in claim 5.
Claim 15
Dependent Claim
172 words
- The process of claim 13, wherein the synthetic gene obtained is: ATC CTT TCC TTG GAT AAA AGA GGT CCG CAA GCT TTG TGT TAG GAA AGG AAC CTA TTT TCT CCA GGC GTT CGA AAC ACA GGT GCT GAG CTC GTT GAC GCT CTG CAG TTC GTT TGC CCA CGA CTC GAG CAA CTG CGA GAC GTC AAG CAA ACG GGT GAC CGC GGT TTC TAC TTC AAC AAA CCG ACT GGT TAC CCA CTG GCG CCA AAG ATG AAG TTG TTT GGC TGA CCA ATG GGT TCT TCT TCT AGA CGT GCT CCG CAG ACT GGT ATC GTT CCA AGA AGA AGA TCT GCA CGA GGC CTC TGA CCA TAG CAA GAT GAA TGC TGC TTC AGA TCT TGT GAC CTG CGT CGT CTA CTT ACG ACG AAG TCT AGA AGA CTG GAC GCA GCA CTC GAG ATG TAC TGC GCA CCG CTG AAA CCG GCT AAA TCT GAG CTC TAC ATG ACG CGT GGC GAC TTT GGC CGA TTT AGA GCT TGA TAA CTCG CGA ACT ATT GAGCCTAG
Claim 16
Dependent Claim
21 words
A process according to claim 8, for the preparation of the synthetic gene encoding for the polypeptide obtained in claim 6.
Claim 17
Dependent Claim
172 words
- The process of claim 15, wherein the synthetic gene obtained is: ATC CTT TCC TTG GAT AAA AGA GGT CCG CAA GCT TTG TGT TAG GAA AGG AAC CTA TTT TCT CCA GGC GTT CGA AAC ACA GGT GCT GAG CTC GTT GAC GCT CTG TAC CTC GTT TGC CCA CGA CTC GAG CAA CTG CGA GAC ATG GAG CAA ACG GGT GAC CGC GGT TTC TAC TTC AAC AAA CCG ACT GGT TAC CCA CTG GCG CCA AAG ATG AAG TTG TTT GGC TGA CCA ATG GGT TCT TCT TCT AGA CGT GCT CCG CAG ACT GGT ATC GTT CCA AGA AGA AGA TCT GCA CGA GGC GTC TGA CCA TAG CAA GAT GAA TGC TGC TTC AGA TCT TGT GAC CTG CGT CGT CTA CTT ACG ACG AAG TCT AGA AGA CTG GAC GCA GCA CTC GAG ATG TAC TGC GCA CCG CTG AAA CCG GCT AAA TCT GAG CTC TAC ATG ACG CGT GGC GAC TTT GGC CGA TTT AGA GCT TGA TAA GTCG CGA ACT ATT CAGCCTAG.
Claim 18
Dependent Claim
30 words
- A method for the promotion of lactation in animals which comprises administering to a lactating animal the synthetic polypeptide analog of hIGF-I obtained through the process of claim 1.
Claim 19
Dependent Claim
30 words
- A method for promoting growth and feed efficiency in animals which comprises administering to such animals the synthetic polypeptide analog of hIGF-I obtained through the process of claim 1.
Claim 20
Dependent Claim
35 words
- A method of increasing the lean and decreasing the fat content of meat producing animals which comprises adminstering to such animals the synthetic polypeptide analog of hIGF-I obtained through the process of claim 1.
References Cited (Backward Citations)
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