METHODS AND MODELS FOR ASSESSING ANTI-AGING BENEFITS OF AGENTS

• US 20130337087A1
• Filed: 06/17/2013
• Published: 12/19/2013
• Est. Priority Date: 06/18/2012
• Status: Active Grant

• Current Assignee: The Procter & Gamble Company
• Original Assignee: The Procter & Gamble Company
• Inventors: OSBORNE, Rosemarie, FINLAY, Deborah Ruth, BASCOM, Charles Carson
• Granted Patent: US 9,593,363 B2
• Time in Patent Office: Days
• US Class Current: 424/725
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

A screening method for identifying an agent as effective for providing an anti-aging skin benefit is provided. The screening method includes culturing first and second human skin samples from about 3 days to about 19 days, wherein the first and second human skin samples are derived from a human skin surgical waste tissue. The first human skin sample is contacted with at least one test agent. The second human skin sample is contacted with a positive control. Transcriptional profiles are generated from the first and second human skin samples, and the at least one test agent is identified as effective for providing an anti-aging skin benefit when the transcriptional profiles of at least two genes show a similar directional change in comparison to one or more untreated control tissue samples.

Claims

1. A screening method for identifying an agent as effective for providing an anti-aging skin benefit, comprising:

a. culturing first and second human skin samples for about 3 days to about 19 days, wherein the first and second human skin samples each comprise an epidermal layer and a dermal layer and are derived from human skin tissue;

b. contacting the first human skin sample with the at least one test agent;

c. contacting the second human skin sample with a positive control;

d. generating a transcriptional profile from the first and second human skin samples, wherein the transcriptional profile of the first human skin sample comprises data related to transcription of at least two genes listed in FIG. 3 and the transcriptional profile of the second human skin sample comprises data related to transcription of the at least two genes; and

e. identifying the test agent as effective for providing an anti-aging skin benefit when the transcriptional profile of the first human skin sample and the transcriptional profile of the second skin sample show similar directional changes for the at least two genes.

2. The screening method of claim 1, wherein the at least two genes comprises from two to nine genes.

3. The screening method of claim 1, wherein the at least two genes are selected from the group consisting of FBN1, FBLN1, TNXB, FN1, LOXL2, COL1A1, COL3A1, ELN, and LOXL1.

4. The screening method of claim 1, wherein contacting the at least one test agent is by addition of the at least one test agent to a culture medium.

5. The screening method of claim 1, wherein the positive control is selected from the group consisting of a vitamin B₃ compound, a peptide, a vitamin A compound, a TGF and combinations thereof.

6. The screening method of claim 1, wherein at least one of the first human skin sample, second human skin sample and control skin sample are obtained from donor tissue.

7. The screening method of claim 6, further comprising repeating steps a) to d), wherein the first and second human skin samples for each repetition are derived from donor tissue from different human donors.

8. The screening method of claim 7, wherein the at least one test agent is different for each repetition.

9. The screening method of claim 7, wherein the positive control induces a directionally appropriate increase or decrease in the expression level of the at least two genes in at least 50% of the second human tissue samples.

10. The screening method of claim 1, further comprising testing the at least one test agent in an assay selected from the group consisting of an enzyme assay, a cell culture assay, or a skin equivalent assay.

11. The screening method of claim 1, wherein the first and second human skin samples are cultured at a relative humidity from about 50% to about 90% and a temperature from about 30°

C. to about 40°

C.

12. The screening method of claim 1, wherein at least one of the first and second human skin samples is cultured on a membrane.

13. A method of manufacturing a cosmetic skin care composition, comprising:

a. indentifying an agent as effective for providing an anti-aging skin benefit according to the screening method of claim 1;

b. combining the agent with a dermatologically acceptable carrier to produce the cosmetic skin care composition; and

c. storing at least a portion of the cosmetic composition in a container.

16. The ex vivo model of claim 13, further comprising a second human skin sample derived from the donor tissue, and comprising an epidermal layer and a dermal layer, the second human skin sample having been cultured in a culture medium at a relative humidity of from about 50% to about 90% and a temperature of from about 30°

C. to about 40°

C. for about 3 days to about 19 days, and having been treated with a test agent.

14. An ex vivo model of an anti-aging skin condition, the model comprising a first human skin sample derived from donor tissue, the first human skin sample comprising an epidermal and dermal layer, and having been cultured in a culture medium for about 3 days to about 19 days, wherein the first human skin sample when treated with a positive control comprising a TGF compound demonstrates a directionally appropriate change in expression level of two or more genes thereby indicating that the donor tissue is acceptable for screening a test agent.

15. The ex vivo model of claim 14, wherein the first and second cultured skin samples are cultured at a relative humidity of from about 50% to about 90% and a temperature of from about 30°

C. to about 40°

C.

17. A screening method for identifying a combination of agents as effective for providing an anti-aging skin benefit, comprising:

a. culturing a first human skin sample and a plurality of second human skin samples from about 3 days to about 19 days, wherein the first human skin sample and the plurality of second human skin samples each comprise an epidermal layer and a dermal layer and are derived from donor tissue;

b. contacting the first human skin sample with a combination of two or more test agents;

c. contacting each of the plurality of second human skin samples with one of the test agents;

d. generating transcriptional profiles from the first human skin sample and the plurality of second human skin samples, wherein the transcriptional profiles comprise data related to transcription of at least two genes listed in FIG. 3; and

e. identifying the combination of test agents as effective for providing an anti-aging skin benefit when the transcriptional profiles of the at least two genes from the first human skin sample shows a synergistic response compared to the transcriptional profiles from the plurality of second human skin samples.

18. The screening method of claim 17, wherein the synergistic response comprises the transcriptional profiles of the two or more genes from the plurality of second human skin samples being regulated in a first direction and the transcriptional profiles of the two or more genes selected from the first human skin sample being regulated in an opposite direction.

19. The screening method of claim 17, wherein the synergistic response comprises the transcriptional profiles of the two or more genes from the first human skin sample being more than or less than the addition of the transcriptional profiles of the two or more genes from the plurality of second human skin samples.

20. The screening method of claim 17, wherein the at least two genes are selected from the group consisting of FBN1, FBLN1, TNXB, FN1, LOXL2, COL1A1, COL3A1, ELN, and LOXL1.