Human insulin-like growth factor analoges with reduced binding to serum carrier proteins and their production in yeast

• US 4,876,242 A
• Filed: 09/21/1987
• Issued: 10/24/1989
• Est. Priority Date: 09/21/1987
• Status: Expired due to Term

• Current Assignee: Merck & Co., Inc.
• Original Assignee: Merck & Co., Inc.
• Inventors: Applebaum, Joy D., Cascieri, Margaret A., Bayne, Marvin L.
• Primary Examiner(s): Phillips, Delbert R.
• Assistant Examiner(s): Chan, Christina
• Application Number:
  US07/099,367
• Time in Patent Office: 764 Days
• Field of Search: 530/303, 530/324, 530/345, 514/3, 514/4, 514/12, 435/172.3
• US Class Current: 514/7.7
• 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 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C07K 14/65 Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

A synthetic gene encoding a 71-amino acid analog of human insulin-like grouwth 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 recepto (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.

Claims

1. A synthetic polypeptide analog of hIGF-I which has the structure:

space="preserve" listing-type="equation">A.sub.1 -A.sub.2 -A.sub.3 -A.sub.4 -LCG-A.sub.5 -A.sub.6 -LV-A.sub.7 AL-A.sub.8 -A.sub.9 -R.sub.1

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

R₁ is VCGDRGFYFNKPTGYGSSSRRAPQTGIVDECCFRSCDLRRLEMYCAPLKPAKSA provided that the A₁ to A₉ groups and R₁ do not constitute GPETLCGAELVDALQF-R₁.

2. The peptide 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.

3. The peptide of claim 1 which is:

FVNQHLCGSHLVEALYL-R₁.

4. The peptide of claim 1 which is:

GPETLCGAELVDALYL-R₁.

5. The peptide of claim 1 which is:

GPQALCGAELVDALQF-R₁.

6. The peptide of claim 1 which is:

GPQALCGAELVDALYL-R₁.

7. The peptide of claim 1 which is:

VNQHLCGSHLVEALYL-R₁.

8. A composition useful for the promotion of lactation, growth and feed efficiency, woulnd healing or stimulating erythropoiesis in animals, or increasing the lean and decreasing the fat content of meat producing animals which comprises an inert carrier and an effective amount of a synthetic polypeptide analog of hIGF-I of claim 1.

9. A method for the promotion of lactation in animals which comprises administering to a lactating animal a synthetic polypeptide analog of hIGF-I of claim 1.

10. A method for promoting growth and feed efficiency in animals which comprises administering to such animals a synthetic polypeptide analog of hIGF-I of claim 1.

11. A method for increasing the lean and decreasing the fat content of meat producing animals which comprises administering to such animals a synthetic polypeptide analog of hIGF-I of claim 1.

12. A method for promoting wound healing in animals which comprises administering to a wounded animal a synthetic polypeptide analog of hIGF-I of claim 1.

13. A method for stimulating erythropoiesis in animals which comprises administering to an animal in need of erythropoiesis a synthetic polypeptide analog of hIGF-I of claim 1.