# US 11,299,548 B2 · Gene combination capable of high-efficiency expression of recombinant human nerve growth factor

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- Number: US 11,299,548 B2
- Document ID: US11299548B2
- Status: Granted
- Kind: B2
- Application number: 16/954,354
- Filed: 2018-11-08
- Published: 2020-12-10
- Granted: 2022-04-12
- Earliest claimed: 2018-03-26
- Assignees: Xintrum Pharmaceuticals, Ltd.
- Applicants: Xintrum Pharmaceuticals, Ltd.
- Inventors: Hai Chen; Hongliang Sun; Yi Zhang; Yuesheng Wang
- Web page: https://patentgrep.com/patent/US11299548B2

## CPC

- C07K 14/48: Nerve growth factor [NGF]
- C07K 16/2875: {against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 (against NGF C07K16/22, against TNF C07K16/241)}
- A61K 38/185: {Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3}
- C07K 2319/02: containing a signal sequence
- C07K 2319/35: containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
- Y02P 20/582: Recycling of unreacted starting or intermediate materials
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

A gene combination capable of high-efficiency expression of rhNGF is provided to optimize gene expression regulation components of a recombinant human nerve growth factor. The gene combination enhances expression of rhNGF. As shown by experiments, the gene combination is capable of high-efficiency expression of a recombinant human nerve growth factor (rhNGF) with natural activity in a eukaryotic expression system.

## Claims

### Claim 1

A human nerve growth factor pro gene nucleic acid molecule, comprising the nucleotide sequence SEQ ID NO: 7.

### Claim 2

A nucleic acid molecule for expression of rhNGF,
wherein the nucleic acid molecule comprises the nucleotide sequence SEQ ID NO: 7, operably linked to the nucleotide sequence of SEQ ID NO: 3 or SEQ ID NO: 5.

### Claim 3 (depends on 2)

A method of producing rhNGF, comprising:
producing rhNGF by expression of the nucleic acid molecule of claim 2.

### Claim 4 (depends on 2)

The nucleic acid molecule of claim 2, further comprising the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2 operably linked to the nucleotide sequence of SEQ ID NO: 3 or SEQ ID NO: 5.

### Claim 5 (depends on 4)

A method of producing rhNGF, comprising:
producing rhNGF by expression of the nucleic acid molecule of claim 4.

### Claim 6

A method of producing rhNGF, comprising:
producing rhNGF by expression of a nucleic acid molecule comprising a nucleotide sequence SEQ ID NO: 3 or SEQ ID NO: 5.