Priority: Dec 20, 2011

(from WO 2013/093407 A1)

• In case you are curious, from Wikipedia: A biological pathway is a series of actions among molecules in a cell that leads to a certain product or a change in a cell. Such a pathway can trigger the assembly of new molecules, such as a fat or protein. Pathways can also turn genes on and off, or spur a cell to move.

• Many factors influence the health and appearance of skin tissue
• Experts have long recognised a list of “active” ingredients that play a key role in skin health.
• The efficacy of an active ingredient relies on its ability to play a specific role in the biological pathway.
• This role is due to its capacity to interact with other molecules in the pathway to induce the desired response.
• Indeed an indirect or direct-Response relationship between an active ingredient and its target allows the ingredient to provide the best effect of a product.

• The genetic era now opens up the possibility to utilise a deeper understanding of genetics in a customised, i.e. person-specific, way.
• A single-nucleotide polymorphism (SNP) found in the target (i.e. molecule to be affected) of an active ingredient (i.e. the key molecule in the drug) provides information on the quality of the expected response
• Use CROSS-VALIDATION of biological target
• This method uses detection of SNP to estimate protein-protein interaction between an ingredient and its direct or indirect target.
• As previously discussed, detecting SNPs to evaluate particular molecule efficacy in a biological pathway is known and is used in pharmacogenetics.
• However, the practice of identifying SNPs associated with an increased or decreased response to an active cosmetic ingredient (ACI) and then testing the individual to determine if they have at least one of said SNPs to determine whether that particularly cosmetic is likely to be effective for that individual, is new.
• The method described herein is different from that used in pharmacogenetics in the sense that, instead of looking at whether a specific SNP is associated with a disorder or defect, the aim is to qualify the effect of an ingredient by querying the target of that ingredient, i.e. by identifying/assessing the presence or absence of SNPs in the targets associated with the active ingredient. By doing so, it is possible to determine if the ingredient will be efficient.

• The skin's health is based upon 6 health categories or “pillars”. Each ingredient relates to one or more of these categories.
• Each ingredient is included in a product in order to take a specific biological pathway in the skin.
• These pathways are: antioxidant pathways for detoxifying, xenobiotic pathways, anti-ageing pathways and skin lightening pathways.
• The ingredient interacts with its biological target inducing a signalling cascade that is responsive of activation of this pathway, for example as shown in FIG. 2.
• The genes have a proposed or established association with the listed ingredients and efficacy outcomes, but the genetic associations are not limited to the skin.

• The ingredient's target corresponds to a protein related to a gene.
• Once the gene is identified, the SNP list for this gene is obtained, for example through NCBI website.
• Calculate SNP impact factor (SIF)
• 5 questions are asked (1: SNP has poor effect on ingredient to 5: SNP has considerable effect on ingredient) to determine SIF.
• A “functional” SNP is one with a SIF of 5 is chosen.

• SNPs are organized into level of interaction with metabolism pathway.

• 1st level: First protein to interact with ingredient (receptor or enzyme responsible for chemical interaction or ingredient modification). Severe consequences on ingredient metabolism if target not functioning.
• 2nd level: Key molecule involved in metabolism (e.g. enzyme within metabolic pathway of ingredient). Moderate consequence.
• 3rd level: Molecule involved in metabolism with minor consequences.

• SNPs that are selected are typically well-represented within the whole population (e.g. greater than 5% frequency within the population).
• However, in some circumstances SNPs may be selected that are very specific to one or more groups within the population (i.e. smaller populations)
• SNPs may be selected that have either a beneficial influence on the metabolism of the ingredient (and therefore its efficacy), or conversely that may have a detrimental influence on the metabolism of the ingredient.

• One of the key aspects of this method is to design specific primers to target the right genotype.
• To amplify DNA, several standard methods can be used such as polymerase chain reaction (PCR), SNAP, or LAPM assay.
• All these techniques are based on the selection of accurate primers.
• Primers were selected according to a number of criteria, including: primer length, the terminal nucleotide in the primer, reasonable GC content and Tm.

• Each ingredient has the ability to be “metabolised” by a person. This ability is based upon the genetic makeup of this person.
• The metabolic pathways of many ingredients are identified and a list is created which details those ingredients that either become inactive due to the presence of a SNP, or a highly beneficial ingredient because the SNP creates a failing or fault that is corrected by this ingredient.
• Once the ingredients have been matched to their target and the associated SNP, this information can be entered into a table or database for future reference.

• It can now be determined if an ingredient is efficient or not when affected by a specific SNP(s) in its target (the aforementioned CROSS-VALIDATION?).
• The final decision reflects the previously determined efficacy of the ingredient.
• If the target is not functional the ingredient will not be recommended. (i.e. customer's SNP not receptive to ingredient)
• In contrast, if the target is not affected by the SNP the ingredient will be recommended (i.e. just a “normal” SNP)
• If the SNP provides again enhanced efficacy of an ingredient, the dosage of this ingredient might be considered before being recommended
• taken into consideration is the genotype identified by the test. This will affect the correlation given on the efficacy of the test

• Niacin (Nicotinic acid, vitamin B3) can act as a ligand that binds and activates certain protein receptors (HM74) sitting in the cell membrane (e.g. G protein-coupled receptors).
• This activation (after some intermediary steps) (see G-protein signaling) causes the release of ATP messengers as part of the cAMP signal pathway (i.e. cAMP is the second messenger).
• In this example the cAMP activate the PKA enzyme which releases its catalytic subunits to phosphorylate (i.e. turn on/off) specific target proteins. (How does PKA know what target to phosphorylate?)
• In this case they activate HSL which promotes free fatty acid (FFA) secretion by adipocyte cells.
• The presence of FFA in the skin may be very helpful as discussed here back in 1957. (Are we ultimately talking about stimulated collagen production here?)
• This is a complicated process, but as far as the patent is concerned the key step is to look at potential mutations in the DNA coding for the receptor protein.
• In particular genes HM74 (protein GPR109B) and HM74A (GPR109A) code for these proteins with the HM74A apparently coding for a better receptor.
• If you identify which of these genes is present (or perhaps even some variation on these genes) you can better predict which customer will better benefit from the metabolic effects of niacin.