# US 11,441,119 B2 · Culture media comprising n-Acyl-X-glutamine dipeptides

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- Number: US 11,441,119 B2
- Document ID: US11441119B2
- Status: Granted
- Kind: B2
- Application number: 16/490,811
- Filed: 2018-03-02
- Published: 2020-01-23
- Granted: 2022-09-13
- Earliest claimed: 2017-03-09
- Assignees: Evonik Operations GmbH
- Applicants: Evonik Operations GmbH
- Inventors: Guenter Knaup; Friedhelm Merz
- Web page: https://patentgrep.com/patent/US11441119B2

## CPC

- C12N 5/0031: {Serum-free culture media}
- C12N 5/0037: {Serum-free medium, which may still contain naturally-sourced components}
- C12N 5/005: {Protein-free medium}
- C12N 5/0682: {Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells (oocytes C12N5/0609)}
- C12N 2500/30: Organic components
- C12N 2500/32: Amino acids
- C12N 2500/90: Serum-free medium, which may still contain naturally-sourced components
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Disclosed herein is a cell culture media containing L-glutamine from a set of N-acylated dipeptides Acyl-X-Q, and L-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource), wherein the variables X, Q, Acyl, R, n(Acyl-X-Q) and n(Qsource) are defined in the general disclosure. Processes of using the cell culture media are also described herein.

## Claims

### Claim 1

A cell culture media, comprising
L-glutamine from a set of N-acylated dipeptides Acyl-XQ; and
L-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource),
wherein:
X is defined as an L-amino acid;
Q is defined as L-glutamine attached via an amide bond to L-amino acid X;
Acyl is defined as a C1-C7-acyl moiety attached via an amide bond to the amino-terminus of L-amino acid X;
R is defined to be in the range of 0.03 to 20;
n(Acyl-X-Q) is the total amount of substance of L-glutamine contained in the set of N-acylated dipeptides Acyl-X-Q in the culture media;
n(Qsource) is the total amount of substance of L-glutamine contained in the set of other glutamine sources Qsource in the culture media;
the constituents of the set of other L-glutamine sources Qsource are selected from the following: Free L-glutamine, dipeptides Y-Q, or mixtures thereof, wherein Y is defined as one of the 20 genetically encoded L-amino acids, wherein Q is defined as L-glutamine attached via an amide bond to L-amino acid Y, and the amide bond connecting Y and Q in dipeptide Y-Q is a regular backbone amide bond involving the carboxy terminus of amino acid Y and the amino terminus of glutamine Q; and
the cell culture media are serum free.

### Claim 2 (depends on 1)

The cell culture media according to claim 1, wherein the cell culture media are chemically defined.

### Claim 3 (depends on 1)

The cell culture media according to claim 1, wherein the only constituent of the set of other L-glutamine sources Qsource is dipeptide Y-Q, wherein Y is Alanine.

### Claim 4 (depends on 1)

The cell culture media according to claim 1, wherein the only constituent of the set of other L-glutamine sources Qsource is free L-glutamine.

### Claim 5 (depends on 1)

The cell culture media according to claim 1, wherein the total concentration of N-acylated dipeptides Acyl-X-Q is in the range of from 1 mM to 8 mM.

### Claim 6 (depends on 1)

The cell culture media according to claim 1, wherein the molar ratio R=n(Acyl-X-Q)/n(Qsource) is defined to be in the range of 0.04 to 15.67.

### Claim 7 (depends on 1)

The cell culture media according to claim 1, wherein the set of N-acylated dipeptides Acyl-X-Q is defined as follows: One or more of acetyl-A-Q, acetyl-G-Q, acetyl-NQ.

### Claim 8 (depends on 1)

The cell culture media according to claim 1, wherein the set of N-acylated dipeptides Acyl-X-Q is defined as follows: One or more of acetyl-A-Q, acetyl-G-Q.

### Claim 9 (depends on 1)

The cell culture media according to claim 1, wherein the set of N-acylated dipeptides Acyl-X-Q is defined as follows: Acetyl-A-Q.

### Claim 10 (depends on 1)

The cell culture media according to claim 1, wherein:
the cell culture media are chemically defined;
the only constituent of the set of other L-glutamine sources Qsource is dipeptide Y-Q, wherein Y is Alanine;
the molar ratio R=n(Acyl-X-Q)/n(Qsource) is defined to be in the range of 0.04 to 15.67;
the set of N-acylated dipeptides Acyl-X-Q is defined as follows: Acetyl-A-Q; and
the concentration of acetyl-A-Q is in the range of from 1 mM to 8 mM.

### Claim 11 (depends on 1)

A process, comprising culturing cells with the cell culture media of claim 1.

### Claim 12 (depends on 1)

A process for manufacturing a cell culture product, the process comprising contacting cells with the cell culture medium of claim 1.

### Claim 13 (depends on 12)

The process according to claim 12, wherein the cells contacted with the cell culture medium are Chinese hamster ovary (CHO) cells.