Insulin analogs and use thereof

Patent metadata

Field	Value
Publication number	US-10017557-B2
Application number	US-201615250459-A
Country	US
Kind code	B2
Filing date	Aug 29, 2016
Priority date	Aug 28, 2015
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

Abstract

Insulin analog with an improved in vitro effect compared with native insulin, a nucleic acid encoding the same, an expression vector including the nucleic acid, a transformant introduced with the expression vector, a method of producing the insulin analog from the transformant, a pharmaceutical composition for treating diabetes containing the insulin analog as an active ingredient, and a method for treating diabetes using the insulin analog or the pharmaceutical composition are described.

First claim

The invention claimed is:

1. A proinsulin peptide comprising an A-chain of SEQ ID NO: 3 indicated in the following General Formula 1 and a B-chain of SEQ ID NO: 4 indicated in the following General Formula 2; wherein an insulin analog composed of the A-chain and the B-chain can be generated from processing of the proinsulin peptide:

General Formula 1

(SEQ ID NO: 3)

Xaa1 -Ile-Val-Glu- Xaa2 -Cys-Cys-Thr-Ser-Ile-Cys-

Xaa3-Leu- Xaa4 -Gln- Xaa5 -Glu-Asn- Xaa6 -Cys- Xaa7

wherein
Xaa1 is glycine;
Xaa2 is glutamine;
Xaa3 is serine;
Xaa4 is alanine, tyrosine, glutamic acid, or asparagine;
Xaa5 is leucine;
Xaa6 is tyrosine; and
Xaa7 is asparagine; and

General Formula 2

(SEQ ID NO: 4)

Phe-Val-Asn-Gln-His-Leu-Cys-Gly-Ser-His-Leu-Val-

Glu-Ala-Leu- Xaa8 -Leu-Val-Cys-Gly-Glu-Arg-Gly-Phe-

Xaa9 -Tyr- Xaa10 - Xaa11 -Lys-Thr

wherein
Xaa8 is tyrosine or glutamic acid;
Xaa9 is phenylalanine, or is absent;
Xaa10 is threonine; and
Xaa11 is proline;
wherein the proinsulin peptide is not a peptide comprising the A-chain of SEQ ID NO: 1 and the B-chain of SEQ ID NO: 2.

2. The proinsulin peptide of claim 1 , wherein, in the B-chain of SEQ ID NO: 4, Xaa8 is tyrosine, and Xaa9 is absent.

3. The proinsulin peptide of claim 1 , wherein, in the A-chain of SEQ ID NO: 3, Xaa4 is glutamic acid or asparagine, and
in the B-chain of SEQ ID NO: 4, Xaa8 is tyrosine, and Xaa9 is phenylalanine.

4. The proinsulin peptide of claim 3 , wherein the peptide comprises the amino acid sequence represented by SEQ ID NO: 16 or 18.

5. The proinsulin peptide of claim 1 ,
wherein, in the A-chain of SEQ ID NO: 3, Xaa4 is glutamic acid and
in the B-chain of SEQ ID NO: 4, Xaa8 is tyrosine, and Xaa9 is absent.

6. The proinsulin peptide of claim 1 , wherein, in the A-chain of SEQ ID NO: 3, Xaa4 is alanine, and
in the B-chain of SEQ ID NO: 4, Xaa8 is glutamic acid, and Xaa9 is absent.

7. The proinsulin peptide of claim 1 , wherein the peptide comprises the amino acid sequence represented by SEQ ID NO: 20 or 22.

8. An insulin analog consisting essentially of an A-chain of SEQ ID NO: 3 indicated in the following General Formula 1 and a B-chain of SEQ ID NO: 4 indicated in the following General Formula 2:

General Formula 1

(SEQ ID NO: 3)

Xaa1-Ile-Val-Glu-Xaa2-Cys-Cys-Thr-Ser-Ile-Cys-

Xaa3-Leu-Xaa4-Gln-Xaa5-Glu-Asn-Xaa6-Cys-Xaa7

wherein
Xaa1 is glycine;
Xaa2 is glutamine;
Xaa3 is serine;
Xaa4 is alanine, tyrosine, glutamic acid, or asparagine;
Xaa5 is leucine;
Xaa6 is tyrosine; and
Xaa7 is asparagine; and

General Formula 2

(SEQ ID NO: 4)

Phe-Val-Asn-Gln-His-Leu-Cys-Gly-Ser-His-Leu-Val-

Glu-Ala-Leu-Xaa8-Leu-Val-Cys-Gly-Glu-Arg-Gly-

Phe-Xaa9-Tyr-Xaa10-Xaa11-Lys-Thr

wherein
Xaa8 is tyrosine or glutamic acid;
Xaa9 is phenylalanine, or is absent;
Xaa10 is threonine; and
Xaa11 is proline;
wherein the insulin analog is not a peptide comprising the A-chain of SEQ ID NO: 1 and the B-chain of SEQ ID NO: 2.

9. The insulin analog of claim 8 , wherein, in the B-chain of SEQ ID NO: 4, Xaa8 is tyrosine, and Xaa9 is absent.

10. The insulin analog of claim 8 ,
wherein, in the A-chain of SEQ ID NO: 3, Xaa4 is glutamic acid or asparagine, and
in the B-chain of SEQ ID NO: 4, Xaa8 is tyrosine, and Xaa9 is phenylalanine.

11. The insulin analog of claim 10 , wherein the peptide comprises the amino acid sequence represented by SEQ ID NO: 16 or 18.

12. The insulin analog of claim 8 , wherein, in the A-chain of SEQ ID NO: 3, Xaa4 is glutamic acid, and in the B-chain of SEQ ID NO: 4, Xaa8 is tyrosine, and Xaa9 is absent.

13. The insulin analog of claim 8 , wherein, in the A-chain of SEQ ID NO: 3, Xaa4 is alanine, and in the B-chain of SEQ ID NO: 4, Xaa8 is glutamic acid, Xaa9 is absent.

14. The insulin analog of claim 8 , wherein the peptide comprises the amino acid sequence represented by SEQ ID NO: 20 or 22.

15. A pharmaceutical composition for treating diabetes comprising the insulin analog according to claim 8 and a pharmaceutically acceptable carrier.

Assignees

• Hanmi Pharm Ind Co Ltd

Inventors

• Kim Jin Young
• Oh Euh Lim
• Lee Jong Soo
• Lim Hyung Kyu
• Choi In Young
• Kwon Se Chang

Classifications

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61K38/00

  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 title

• C07K14/62 Primary

  Insulins · CPC title

• C12N15/70

  Vectors or expression systems specially adapted for E. coli · CPC title

• C07K1/12

  by hydrolysis {, i.e. solvolysis in general} · CPC title