BLENDED FORMULATIONS

Abstract

The present invention relates to a formulation comprising blends of formulations (or colloidal dispersions) and its topical application. The formulation comprises at least two different types of colloidal dispersion comprising deformable colloidal particles, wherein the deformable colloidal particles comprise a non-ionic surfactant and/or a phospholipid. The deformable colloidal particles of the invention may comprise an agent of interest (AOI) or may be free of an AOI. The formulation may comprise an AOI that is not associated with the deformable colloidal particles. The present invention also includes kits comprising the formulation of the present invention and the use of the formulation in medicine, skin care and cosmetics.

Claims

1. A formulation comprising a first colloidal dispersion and a second colloidal dispersion, wherein each colloidal dispersion comprises deformable colloidal particles comprising a surfactant and optionally a phospholipid, and wherein one or more of the deformable colloidal particles of the first colloidal dispersion comprise a modified component comprising a first agent of interest (AOI), wherein the modified component is a lipid or surfactant bonded to an AOI, such that the entire AOI is outside the particle membrane, and wherein the first AOI is zinc.

2. The formulation of claim 1, wherein one or more of the deformable colloidal particles of the second colloidal dispersion comprise a second AOI and wherein the first AOI and the second AOI are different.

3. The formulation of claim 1, wherein one or more of the deformable colloidal particles of the first colloidal dispersion and/or one or more of the deformable colloidal particles of the second colloidal dispersion comprise one or more further AOIs and wherein each AOI is different.

4. The formulation of claim 1, wherein the formulation comprises one or more further colloidal dispersions comprising deformable colloidal particles comprising a surfactant and optionally a phospholipid and wherein one or more of the particles of at least two of the dispersions comprise one or more AOIs and wherein the first AOI, second AOI and each further AOI is different.

5. through 9. (canceled)

10. The formulation of claim 1, comprising both a modified surfactant component and a modified lipid component, optionally wherein the bond between each AOI and the lipid or surfactant of each modified component is a covalent bond.

11. (canceled)

12. The formulation of claim 2, wherein the second AOI is selected from the group consisting of an element, an ion, an inorganic salt, a small molecule, an amino acid, a peptide, a protein, a carbohydrate, a lipid, a micronutrient, a macromolecule or a macrocyclic molecule.

13. The formulation of claim 2, wherein the second AOI is selected from the group consisting of zinc, ascorbate, tetrapeptide, tripeptide, salicylic acid, Vitamin D, tocopherol or menthol.

14. (canceled)

15. The formulation of claim 1, wherein the formulation comprises chlorhexidine, salicylic acid, capsaicin, caffeine or tocopherol, optionally wherein the chlorhexidine, salicylic acid, capsaicin, caffeine or tocopherol is not associated with the deformable colloidal particles.

16. (canceled)

17. The formulation of claim 1, wherein the formulation comprises at least three colloidal dispersions.

18. The formulation of claim 1, wherein the formulation comprises at least four colloidal dispersions.

19. The formulation of claim 1, wherein the formulation is topically applied.

20. The formulation of claim 1, wherein the formulation is applied weekly, between weekly and daily, daily, twice daily, three times a day or four times a day.

21. The formulation of claim 1, wherein the lipid is phosphatidylcholine.

22. The formulation of claim 1, wherein the surfactant is polysorbate 80.

23. The formulation of claim 1, wherein the formulation further includes one or more buffers, chelators, humectants, lubricants, antioxidants, preservatives, microbicides, antimicrobials, emollients, co-solvents or thickeners.

24. through 36. (canceled)

37. A method of cosmetically improving the appearance of a subject, the method comprising topically applying the formulation of claim 1 to the skin of a subject.

38. through 40. (canceled)

41. A method of making the formulation of claim 1.

42. A method of treating or preventing a disease in a subject comprising topically applying the formulation of claim 1 to the subject, wherein the disease is treated, optionally wherein the disease is acne, dry skin, itchy skin, red skin, irritated skin, scaly skin, ageing skin, thinning skin, uneven skin tone, periorbital skin, photo-aged skin or dermatitis.

43. (canceled)

44. A method of delivering an AOI to or through the skin of a patient, the method comprising topically applying to the skin of the patient the formulation of claim 1 in an amount sufficient to penetrate the skin to deliver the AOI.

45. A kit comprising one or more compartments, wherein at least one compartment contains the formulation of claim 1, wherein the kit comprises instructions for administrating the formulation to a subject in need thereof.

46. (canceled)

47. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0264] The invention is described below with reference to the following examples and figures in which:

[0265] FIG. 1 shows the structure of a colloidal particle (vesicle or Sequessome™/Transfersome™/Tethersome) of the invention. The spherical bilayer may comprise a surfactant or a lipid or a surfactant and a lipid. As can be seen from this diagram, the bilayer is two molecules wide;

[0266] FIG. 2 shows a typical formulation in accordance with the first aspect of the invention. This formulation comprises a colloidal dispersion (or Sequessome™/Transfersome™/Tethersome intermediate) comprising colloidal particles, a colloidal dispersion comprising colloidal particles comprising an agent of interest (AOI) (e.g. a peptide, ascorbic acid or zinc) and an AOI that is not associated with the colloidal particles (e.g. chlorhexidine or capsaicin);

[0267] FIG. 3 shows a table summarising Example Formulations 1 to 4;

[0268] FIG. 4 shows the change in sebum levels compared to baseline level on day 0 (=100%);

[0269] FIG. 5 shows the change in percentage of comedones compared to day 0 occurrence (100%);

[0270] FIG. 6 shows the change in percentage incidence of pustules compared to day 0 occurrence (100%);

[0271] FIG. 7 shows the reduction in measured wrinkle volume and the reduction in appearance of fine lines and wrinkles after treatment with a two types of formulation in accordance with the present invention;

[0272] FIG. 8 shows a comparison of reduction in wrinkle volume (as a percentage of Week 0 baseline volume) after treatment with a formulation in accordance with the present invention, a UK leading own brand (in-market product) and a control;

[0273] FIG. 9 shows a comparison of reduction in perceived appearance of wrinkles and fine lines (as a percentage of Week 0 baseline volume) after treatment with a formulation in accordance with the present invention, a UK leading own brand (in-market product) and a control; and

[0274] FIG. 10 shows a comparison of reduction in appearance of pen-orbital dark circles (as a percentage of Week 0 baseline volume) after treatment with a formulation in accordance with the present invention, a UK leading own brand (in-market product) and a control.

EXAMPLES

Example 1: Method of Manufacture

[0275] The simplified generic manufacturing process of formulations in accordance with the present invention is as follows. Firstly, the colloidal dispersions comprising the deformable colloidal particles (for example, the Sequessome™ intermediate, Transfersome™ intermediate or Tethersome intermediate) are made. These colloidal dispersions are each formed from an ‘organic phase’ containing alcohol-soluble components and an ‘aqueous phase’ consisting of water-soluble components. Secondly, the gel phase (a thickener) is formed within which the colloidal dispersion(s) are dispersed. During this secondary stage, more than one kind of colloidal dispersion is introduced (each kind containing differing AOIs (or ‘extra’ ingredients) associated with the colloidal particles to give them unique characteristics), such that the final gelled product is then a blend of one or more individual kinds of vesicles (or colloidal particles). The AOIs or ‘extras’ may include zinc stearate, palmitoyl peptides, palmitoyl ascorbic acid (PAA), myristyl salicylate, tridecyl salicylate, vitamin D or vitamin E etc. (the ‘tethered’ ingredients). If desired, an AOI not associated with the colloidal particles can be added to the gel phase during secondary manufacture. If desired the first phase of the manufacturing process can be ordered so that the colloidal dispersions comprising the colloidal particles without any AOI are made in one or more formulations and then to each formulation an AOI (attached to a [phospho]lipid or a surfactant) is added. The second gel phase then occurs as above.

Example 2: Example Formulations

Example Formulation 1

[0276]

TABLE-US-00004 Quantity Required Per 100 g Final Product (g) Organic Phase - 1600 ppm PAA Intermediate SPC 4.12200 Ethanol 2.19660 BHT 0.01200 Methyl-4-hydroxybenzoate 0.15000 Ethyl-4-hydroxybenzoate 0.15000 Benzyl alcohol 0.31500 Polysorbate 80 0.40800 PAA 0.09600 Linalool 0.06000 Organic Phase Sub-Total 7.50960 Aqueous Phase - 1600 ppm PAA Intermediate Sodium metabisulphite 0.03000 Citric acid 0.04320 Na EDTA 0.06000 diNa hydrogen phos 12H2O 0.17280 Water 27.18420 Aqueous Phase Sub-Total 27.49020 PAA Tethersome Intermediate Total 34.99980 Organic Phase - 300 ppm Tetrapeptide Intermediate SPC 1.37400 Ethanol 0.74120 BHT 0.0040 Methyl-4-hydroxybenzoate 0.05000 Ethyl-4-hydroxybenzoate 0.05000 Benzyl alcohol 0.10500 Polysorbate 80 0.15300 Palmitoyl peptide 0.00600 Linalool 0.02000 Organic Phase Sub-Total 2.50320 Aqueous Phase - 300 ppm Tetrapeptide Intermediate Sodium metabisulphite 0.01000 Citric acid 0.01440 Na EDTA 0.02000 diNa hydrogen phos 12H2O 0.05760 Water 9.06140 Aqueous Phase Sub-Total 9.16340 Tetrapeptide Tethersome Intermediate Total 11.66660 Organic Phase - 300 ppm Tripeptide Intermediate SPC 1.37400 Ethanol 0.74120 BHT 0.00400 Methyl-4-hydroxybenzoate 0.05000 Ethyl-4-hydroxybenzoate 0.05000 Benzyl alcohol 0.10500 Polysorbate 80 0.15300 Palmitoyl peptide 0.00600 Linalool 0.02000 Organic Phase Sub-Total 2.50320 Aqueous Phase - 300 ppm Tripeptide Intermediate Sodium metabisulphite 0.01000 Citric acid 0.01440 Na EDTA 0.02000 diNa hydrogen phos 12H2O 0.05760 Water 9.06140 Aqueous Phase Sub-Total 9.16340 Tripeptide Tethersome Intermediate Total 11.66660 Sum of Tethersome Intermediates 58.33300 Gel Phase - Final Product Sodium hydroxide 0.11300 Carbopol 974P 0.75000 Glycerol 3.00000 Citric acid 0.05600 diNa hydrogen phos 12H2O 0.24200 Water 37.50600 Gel Phase Sub-Total 41.66700 Overall Total 100.00000

Example Formulation 2

[0277]

TABLE-US-00005 Quantity Required Per 100 g Final Product (g) Organic Phase - 1600 ppm PAA Intermediate SPC 2.06100 Ethanol 1.09830 BHT 0.00600 Methyl-4-hydroxybenzoate 0.07500 Ethyl-4-hydroxybenzoate 0.07500 Benzyl alcohol 0.15750 Polysorbate 80 0.20400 PAA 0.04800 Linalool 0.03000 Organic Phase Sub-Total 3.75480 Aqueous Phase - 1600 ppm PAA Intermediate Sodium metabisulphite 0.01500 Citric acid 0.02160 diNa hydrogen phos 12H2O 0.08640 Water 13.62210 Aqueous Phase Sub-Total 13.74510 PAA Tethersome Intermediate Total 17.49990 Organic Phase - 300 ppm Tetrapeptide Intermediate SPC 0.68700 Ethanol 0.37060 BHT 0.00200 Methyl-4-hydroxybenzoate 0.02500 Ethyl-4-hydroxybenzoate 0.02500 Benzyl alcohol 0.05250 Polysorbate 80 0.07650 Palmitoyl peptide 0.00300 Linalool 0.01000 Organic Phase Sub-Total 1.25160 Aqueous Phase - 300 ppm Tetrapeptide Intermediate Sodium metabisulphite 0.00500 Citric acid 0.00720 diNa hydrogen phos 12H2O 0.02880 Water 4.54070 Aqueous Phase Sub-Total 4.58170 Tetrapeptide Tethersome Intermediate Total 5.83330 Organic Phase - 300 ppm Tripeptide Intermediate SPC 0.68700 Ethanol 0.37060 BHT 0.00200 Methyl-4-hydroxybenzoate 0.02500 Ethyl-4-hydroxybenzoate 0.02500 Benzyl alcohol 0.05250 Polysorbate 80 0.07650 Palmitoyl peptide 0.00300 Linalool 0.01000 Organic Phase Sub-Total 1.25160 Aqueous Phase - 300 ppm Tripeptide Intermediate Sodium metabisulphite 0.00500 Citric acid 0.00720 diNa hydrogen phos 12H2O 0.02880 Water 4.54070 Aqueous Phase Sub-Total 4.58170 Tripeptide Tethersome Intermediate Total 5.83330 Organic Phase - Zinc Stearate Intermediate SPC 3.43500 Ethanol 1.82850 BHT 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.34000 Zinc Stearate 0.08200 Linalool 0.05000 Organic Phase Sub-Total 6.25800 Aqueous Phase - Zinc Stearate Intermediate Sodium metabisulphite 0.02500 Citric acid 0.03600 diNa hydrogen phos 12H2O 0.14400 Water 22.70350 Aqueous Phase Sub-Total 22.90850 Zinc Tethersome Intermediate Total 29.16650 Sum of Tethersome Intermediates 58.33300 Gel Phase - Final Product Sodium hydroxide 0.11300 Carbopol 974P 0.75000 Glycerol 3.00000 Citric acid 0.05600 diNa hydrogen phos 12H2O 0.24200 Water 37.50600 Gel Phase Sub-Total 41.66700 Overall Total 100.00000

Example Formulation 3

[0278]

TABLE-US-00006 Quantity Required Per 100 g Final Product (g) Organic Phase - Empty Sequessome Intermediate SPC 3.57300 Ethanol 1.75000 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.23600 Linalool 0.05000 Organic Phase Sub-Total 6.13150 Aqueous Phase - Empty Sequessome Intermediate Na dihydrogen phos 2H2O 0.22200 diNa hydrogen phos 12H2O 0.38600 Water 34.02100 Aqueous Phase Sub-Total 34.62900 Empty Sequessome Intermediate Total 40.76050 Organic Phase - Zinc Stearate Intermediate SPC 3.57300 Ethanol 1.66800 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.23600 Zinc stearate 0.08200 Linalool 0.05000 Organic Phase Sub-Total 6.13150 Aqueous Phase - Zinc Stearate Intermediate Na dihydrogen phos 2H2O 0.22200 diNa hydrogen phos 12H2O 0.38600 Water 34.02100 Aqueous Phase Sub-Total 34.62900 Zinc Tethersome Intermediate Total 40.76050 Sum of Sequessome and 81.52100 Tethersome lntermediates Gel Phase - Final Product Sodium hydroxide 0.30000 Carbopol 974P 1.00000 Glycerol 3.00000 Water 14.17900 Gel Phase Sub-Total 18.47900 Overall Total 100.00000

Example Formulation 4

[0279]

TABLE-US-00007 Quantity Required Per 100 g Final Product (g) Organic Phase - Empty Sequessome Intermediate Soy phosphatidylcholine (SPC) 3.43500 Ethanol 1.82550 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.42500 Linalool 0.05000 Organic Phase Sub-Total 6.25800 Aqueous Phase - Empty Sequessome Intermediate Citric acid monohydrate 0.03600 diNa hydrogen phos 12H2O 0.14400 Water 22.72850 Aqueous Phase Sub-Total 22.90850 Empty Sequessome Intermediate Total 29.16650 Organic Phase - Zinc Stearate Intermediate Soy phosphatidylcholine SPC 3.43500 Ethanol 1.74350 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.42500 Zinc stearate 0.08200 Linalool 0.05000 Organic Phase Sub-Total 6.25800 Aqueous Phase - Zinc Stearate Intermediate Citric acid monohydrate 0.03600 diNa hydrogen phos 12H2O 0.14400 Water 22.72850 Aqueous Phase Sub-Total 22.90850 Zinc Tethersome Intermediate Total 29.16650 Sum of Sequessome and 58.33300 Tethersome Intermediates Gel Phase - Final Product Citric acid monohydrate 0.05600 diNa hydrogen phos 12H2O 0.24200 Methyl cellulose (4000 cPs, 88,000 Mw) 1.75000 Glycerol 3.00000 Water 36.00600 Gel Phase Sub-Total 40.16700 Chlorhexidine digluconate 1.50000 Overall Total 100.00000

Example 3

[0280] An in-use study in healthy volunteers to investigate the anti-spot efficacy of two Example Formulations comprising chlorhexidine, using objective instrumental assessments of skin sebum levels and subjective visual assessments of lesion prevalence, against a placebo following a 3-week use period.

[0281] The effectiveness of Formulation 4 and a second formulation (“Formulation 4.2”) was compared with a control product in their ability to control sebum and reduce the incidence of acne in acne-prone individuals.

[0282] The Example Formulations (n=36 and n=37) contained empty Sequessomes™ and Tethersomes with tethered zinc and other excipients, including chlorhexidine as an anti-microbial preservative. The Sequessomes™ in the two Formulations contained different ratios of SPC:Tween (polysorbate) to assess if this affected the efficacy of the product. The control product contained no vesicles, zinc or chlorhexidine (n=20).

[0283] Data Review

[0284] The study assessed the efficacy and safety of treatment with Formulation 4 and Formulation 4.2 compared with placebo (control) in healthy volunteers with oily and spot-prone skin, confirmed by a clinical trials assistant prior to enrolment.

[0285] Volunteers were either assessed at the centre on Day 3, 7, 14 and 21 or purely provided their own subjective assessments from home.

[0286] In total, 224 volunteers completed the 3-week study: 37 were allocated to Formulation 4, 36 were allocated to Formulation 4.2 and 20 to placebo. Objective assessments included measurement of sebum on the face, Visia CR Photography (used to assess pore size and skin smoothness) and lesion counts (comedones, microcysts, papules and pustules). The subjective assessment was carried out using a subject Self Perception Questionnaire (SPQ).

[0287] Sebum levels were reduced by over 20% from Baseline after 1 week of Formulation 4 and by more than 50% after 3 weeks' treatment (see FIG. 4). Published data demonstrate that a reduction in sebum production of 30-50% correlates with reduced acne symptoms..sup.1 The number of comedones (blocked pores) was also reduced with Formulation 4: by 50% from Baseline after 1 week and by more than 80% after 3 weeks (see FIG. 5). Notably, the number of pustules was reduced by 90% from Baseline after 1 week and they were virtually cleared after 3 weeks' treatment (see FIG. 6).

[0288] These objective measurements were further validated by the subjective assessments. Additional subjects were given formulations to try and questionnaires to fill in, such that the sample size for each of the groups rose to 102 subjects, of which: [0289] 82% reported less painful spots [0290] 80% said they had less shiny skin [0291] 76% felt their blackheads reduced [0292] 75% reported less swollen spots [0293] 75% found they had no spot recurrence.

[0294] Formulation 4 was well tolerated with no AEs reported and no erythema at the test site.

[0295] This research demonstrates that Formulation 4 is a very effective treatment for acne. The reduction in sebum and both spots and blackheads at 21 days also support the role of Formulation 4 in preventing future recurrence of the symptoms.

[0296] As mentioned, the objectively measured results demonstrated that against the control product the test products: [0297] Significantly reduced surface sebum (by up to 50%); [0298] Significantly reduced Comedones; [0299] Significantly reduced the numbers of papules and pustules.

TABLE-US-00008 TABLE 4 summary of results Results of objective assessment Formulation 4.2 Formulation 4 Reduction in sebum after. . . 1 week >20%; 1 week >20%; 3 weeks >50% 3 weeks >50% Reduction in comedones after. . . 1 week >40%; 1 week = 50%; 3 weeks >90% 3 weeks >80% Reduction in pustules after. . . 1 week = 90%; 1 week >90%; 3 weeks >98% 3 weeks >98% Results of subjective assessment: % agreeing/strongly agreeing Product reduced blackheads 64.7% 76.5% Product stopped spots recurring 69.6% 74.5% Spots less painful (tender to touch) 66.7% 82.4% Spots not as swollen 73.5% 75.5% Skin less shiny 59.8% 80.4% Skin less oily 67.6% 77.5% Skin less greasy 69.6% 75.5%

[0300] These results show the multifunctional nature of the formulation. It is believed that the colloidal particles in the formulations both assist in the clearing of the excess sebum from the skin surface, whilst their penetration into the skin drags the chlorhexidine in solution into the skin structure, killing bacteria. The tethered zinc may also have had a role in both reducing sebum production and/or having an anti-microbial function.

[0301] Methods

TABLE-US-00009 TABLE 5 Summary Protocol Title: An in-use study in healthy volunteers to investigate the anti-spot efficacy of two test formulations, using objective instrumental assessments of skin sebum levels and subjective visual assessments of lesion prevalence, against a placebo following a 3-week use period. Study design: Single-blind, within-subject comparison. Test Groups: Formulation 4.2. CBL-DERM-14-005-E Formulation 4. CBL-DERM-006-F The two test formulations were identical apart from containing colloidal particles that had slightly different SPC:Tween ratios Control: placebo. CBL-DERM-14-007/8-P Dose regime: The test groups followed a 12-week usage schedule according to treatment specific in-use regimes. Duration of 21 Days. study: Number of 36 subjects completed the active phase for group 1, 37 subjects for subjects: group 2 and 20 subjects completed assessments for group 3. Duration of Study Started: w/c 26, Jan. 2015 study: Study Ended: w/e 20, Feb. 2015 Location: Princeton Consumer Research Ltd. 307 College Road East Princeton New Jersey 08540

[0302] Test Formulations

[0303] 1. FORMULATION 4.2 AND FORMULATION 4 (CBL-DERM-14-005-E and CBL-DERM-006-F)

[0304] SOY PHOSPHATIDYLCHOLINE

[0305] BUTYLHYDROXYANISOLE

[0306] ETHANOL

[0307] ZINC STEARATE

[0308] METHYL-4-HYDROXYBENZOATE

[0309] ETHYL-4-HYDROXYBENZOATE

[0310] BENZYLALCOHOL

[0311] POLYSORBATE 80

[0312] CHLORHEXIDINE DIGLUCONATE

[0313] CITRIC ACID MONOHYDRATE

[0314] DI-SODIUM HYDROGEN ORTHOPHOSPHATE ANHYDROUS

[0315] SODIUM HYDROXIDE

[0316] CARBOPOL 974P

[0317] GLYCEROL

[0318] Linalool

[0319] WATER

[0320] The quantity of each component used is shown in the tables below. Formulation 4 (CBL-DERM-14-006-F) was the same as the exemplary composition set out in Example Formulation 4, above, except that Formulation 4 (CBL-DERM-14-006-F) contains carbopol instead of methyl cellulose (at the same quantity) and 0.11300 g of sodium hydroxide (the amount of water in the gel phase is adjusted accordingly).

[0321] 2. CONTROL (CBL-DERM-14-007/8-P)

[0322] METHYL-4-HYDROXYBENZOATE

[0323] ETHYL-4-HYDROXYBENZOATE

[0324] CITRIC ACID MONOHYDRATE

[0325] DI-SODIUM HYDROGEN ORTHOPHOSPHATE DODECAHYDRATE

[0326] SODIUM HYDROXIDE

[0327] CARBOPOL 974P (CARBOMER)

[0328] WATER

[0329] Summary Formulation Information

Summary of Formulation 4 (CBL-DERM-14-006-F)

[0330]

TABLE-US-00010 Quantity Required Per Ingredient 100 g Final Product (g) SPC (dry mass) 6.870 Polysorbate 80 0.850 Benzylalcohol 0.525 Methyl-4-hydroxybenzoate 0.250 Ethyl-4-hydroxybenzoate 0.250 Butylhydroxyanisol 0.020 Linalool 0.100 Disodium hydrogen phosphate 12 H.sub.2O 0.530 Citric acid monohydrate 0.128 Glycerol 3.000 Ethanol 3.569 Sodium hydroxide 0.113 Carbopol 974P NF 0.750 Water 81.463 Agent of interest - tethered to Tethersomes Zinc stearate 0.082 Agent of interest - present in continuous phase Chlorhexidine digluconate (20% solution) ** 1.5 pH 5.5 ** % with respect to the salt

Summary of Formulation 4.2 (CBL-DERM-14-005-E)

[0331]

TABLE-US-00011 Quantity Required Per Ingredient 100 g Final Product (g) SPC (dry mass) 7.146 Polysorbate 80 0.472 Benzylalcohol 0.525 Methyl-4-hydroxybenzoate 0.250 Ethyl-4-hydroxybenzoate 0.250 Butylhydroxyanisol 0.020 Linalool 0.100 Disodium hydrogen phosphate 12 H.sub.2O 0.530 Citric acid monohydrate 0.128 Glycerol 3.000 Ethanol 3.418 Sodium hydroxide 0.113 Carbopol 974P NF 0.750 Water 81.716 Agent of interest - tethered to Tethersomes Zinc stearate 0.082 Agent of interest - present in continuous phase Chlorhexidine digluconate (20% solution) ** 1.5 pH 5.5 ** % with respect to the salt

Summary of Control (CBL-DERM-14-007/8-P)

[0332]

TABLE-US-00012 Quantity Required Per Ingredient 100 g Final Product (g) SPC (dry mass) Polysorbate 80 Benzylalcohol Methyl-4-hydroxybenzoate 0.250 Ethyl-4-hydroxybenzoate 0.250 Butylhydroxyanisol Linalool Disodium hydrogen phosphate 12 H.sub.2O 0.530 Citric acid monohydrate 0.128 Glycerol Ethanol Sodium hydroxide 0.150 Carbopol 974P NF 1.000 Water 97.692 Agent of interest - tethered to Tethersomes Zinc stearate Agent of interest - present in continuous phase Chlorhexidine digluconate (20% solution) ** pH 5.5 ** % with respect to the salt

[0333] Detailed Formulation Information

Formulation 4 (CBL-DERM-14-006-F)

[0334]

TABLE-US-00013 Quantity Required Per 100 g Final Product (g) Organic Phase - Empty Sequessome Intermediate SPC 3.43500 Ethanol 1.82550 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.42500 Linalool 0.05000 Organic Phase Sub-Total 6.25800 Aqueous Phase - Empty Sequessome Intermediate Citric acid monohydrate 0.03600 diNa hydrogen phos 12H2O 0.14400 Water 22.72850 Aqueous Phase Sub-Total 22.90850 Empty Sequessome Intermediate Total 29.16650 Organic Phase - Zinc Stearate Intermediate SPC 3.43500 Ethanol 1.74350 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.42500 Zinc stearate 0.08200 Linalool 0.05000 Organic Phase Sub-Total 6.25800 Aqueous Phase - Zinc Stearate Intermediate Citric acid monohydrate 0.03600 diNa hydrogen phos 12H2O 0.14400 Water 22.72850 Aqueous Phase Sub-Total 22.90850 Zinc Tethersome Intermediate Total 29.16650 Sum of Sequessome and Tethersome 58.33300 Intermediates Gel Phase - Final Product Citric acid monohydrate 0.05600 diNa hydrogen phos 12H2O 0.24200 Sodium hydroxide 0.11300 Carbopol 974P 0.75000 Glycerol 3.00000 Water 36.00600 Gel Phase Sub-Total 40.16700 Chlorhexidine digluconate 1.50000 Overall Total 100.00000

Formulation 4.2 (CBL-DERM-14-005-E)

[0335]

TABLE-US-00014 Quantity Required Per 100 g Final Product (g) Organic Phase - Empty Sequessome Intermediate SPC 3.57300 Ethanol 1.75000 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.23600 Linalool 0.05000 Organic Phase Sub-Total 6.13150 Aqueous Phase - Empty Sequessome Intermediate Citric acid monohydrate 0.06400 diNa hydrogen phos 12H2O 0.26500 Water 34.30000 Aqueous Phase Sub-Total 34.62900 Empty Sequessome Intermediate Total 40.76050 Organic Phase - Zinc Stearate Intermediate SPC 3.57300 Ethanol 1.66800 BHA 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.23600 Zinc stearate 0.08200 Linalool 0.05000 Organic Phase Sub-Total 6.13150 Aqueous Phase - Zinc Stearate Intermediate Citric acid monohydrate 0.06400 diNa hydrogen phos 12H2O 0.26500 Water 34.30000 Aqueous Phase Sub-Total 34.62900 Zinc Tethersome Intermediate Total 40.76050 Sum of Sequessome and Tethersome 81.52100 Intermediates Gel Phase - Final Product Sodium hydroxide 0.11300 Carbopol 974P 0.75000 Glycerol 3.00000 Water 13.11600 Gel Phase Sub-Total 16.97900 Chlorhexidine digluconate 1.50000 Overall Total 100.00000

[0336] Results

[0337] Objectively Measured Parameters

[0338] Sebum Reduction

TABLE-US-00015 TABLE 6 Average reduction in Sebum score from Day 0. A negative score indicates an increase in sebum. n Day 3 Day 7 Day 14 Day 21 Formulation 36 20.47 34.94 71.64 90.11 4.2 Formulation 4 37 14.51 36.30 85.35 98.49 Control 20 −21.30 −48.60 −40.35 −44.95 1 vs. 2 t test p- 0.0163 0.6767 0.2065 0.3936 1 vs. Control values 7.04E−16 5.27E−23 9.74E−11 5.69E−16 2 vs. Control 1.06E−14 1.62E−24 1.10E−14 4.63E−17

[0339] At all time points, both test formulations reduced sebum significantly more than the control at p<0.1%. On day 3 Formulation 4.2 was better than Formulation 4 at p<0.02. There was no significant difference in the effectiveness of the two test formulations at all later time points.

[0340] By Day 21, both test formulations had reduced the average sebum levels to less than 50% of the starting value (see FIG. 4).

[0341] Comedone Reduction

TABLE-US-00016 TABLE 7 Average reduction in occurrence of comedones from Day 0. A positive score indicates an increase in comedones. n Day 3 Day 7 Day 14 Day 21 Formulation 36 −1.44 −2.31 −2.92 −3.44 4.2 Formulation 4 37 −1.14 −1.97 −2.84 −3.38 Control 20 1.20 1.30 1.25 0.35 1 vs. 2 t test p- 0.414 0.468 0.903 0.934 1 vs. Control values 2.35E−06 2.98E−06 4.66E−05 6.63E−04 2 vs. Control 1.08E−07 2.93E−05 7.20E−05 8.97E−05

[0342] At all time points, both test formulations reduced comedones significantly more than the control at p<0.1%. There was no significant difference in the effectiveness of the two test formulations at all time points (see FIG. 5).

[0343] Papule Reduction

TABLE-US-00017 TABLE 8 Analysis of progression participants who had blemishes at day 0 n Day 0 Day 3 Day 7 Day 14 Day 21 Formulation 4.2 5 13 5 1 0 0 Formulation 4 7 16 9 5 0 0 Control 4 10 9 8 5 3

[0344] In those participants who began the trial with at least one papule, all treatments showed a reduction in papule numbers over 21 days.

TABLE-US-00018 TABLE 9 Probabilities of U-statistic (Mann-Whitney) calculated on lesion reductions from Day 0 D3 v D7 vs D14 vs D21 vs D0 D0 D0 D0 1 vs. 2 0.146 0.044 0.373 0.373 1 vs. Control 0.043 0.019 0.056 0.089 2 vs. Control 0.110 0.093 0.110 0.254

[0345] The test statistic (U) shows that Formulation 4.2 was better than Control @<5% on Day 3 and Day 7 and @<10% on Day 14 and Day 21 and better that Formulation 4 @<5% on Day 7. Formulation 4 was better than Control @<10% on Day 7.

[0346] Pustule Reduction

TABLE-US-00019 TABLE 10 Reduction in pustule numbers from day 0. A positive score indicates an increase in numbers of pustules. n Day 3 Day 7 Day 14 Day 21 Formulation 36 −1.28 −1.97 −2.11 −2.17 4.2 Formulation 4 37 −0.92 −1.49 −1.57 −1.59 Control 20 0.60 0.05 0.15 −0.75 1 vs. 2 t test p- 0.127 0.221 0.228 0.226 1 vs. Control values 2.71E−08 2.00E−05 9.67E−05 1.37E−02 2 vs. Control 8.60E−07 7.48E−04 1.04E−03 9.15E−02

[0347] At all time points from Day 3 to Day 14, both test formulations reduced pustule numbers significantly more than the control at p<0.1%. At Day 21, Formulation 4.2 still outperformed control at p<2.0%; Formulation 4 outperformed control at p<10%. There was no significant difference in the effectiveness of the two test formulations at all time points (see FIG. 6).

REFERENCES

[0348] 1. Janiczek-Dolphin N1, Cook J, Thiboutot D, Harness J, Clucas A: Can sebum reduction predict acne outcome? Br J Dermatol. 2010 Oct;163(4):683-8. doi: 10.1111/j. 1365-2133.2010.09878

Example 4: Anti-Ageing Trials. A Summary of Trials and Results

SUMMARY

[0349] Two trials of novel products have been undertaken to assess their efficacy and how they were perceived and received by users. See FIGS. 7, 8, 9 and 10.

[0350] PROAWR1: A home-use study demonstrating the subjective and objective improvements in wrinkles and skin appearance after treatment with two different formulations of the same ingredients used with three different regimes

[0351] PROAWR2: home-use study in three parallel groups of a preferred formulation and regime against a marketed comparator (in-market product A) and a control (placebo) product.

[0352] The test formulas in the two trials were variants of the same ingredients, formulated by blending a mixture of Sequessome-based vesicles (i.e. colloidal particles), some of which had additional moieties tethered to the vesicle membrane.

[0353] Headline Results

[0354] PROAWR1: [0355] 100% of all participants on all three regimes had measured reduction in wrinkle volume after the first week [0356] The average scores both wrinkle volume reduction from baseline measurements and the reduction in appearance of wrinkles from baseline scores were highly significant (p<0.001) from week one [0357] There was mean reduction in actual wrinkle volume of 17.46% at week 12; the mean Glogau (appearance) score had reduced by 25.86% over the same period [0358] In the subjective assessments: [0359] 100% reported that their skin felt smoother and healthier and looked and felt plumper after 8 weeks [0360] Over 90% reported that their skin looked smoother and healthier, felt more elastic and that the bags and circles under their eyes had reduced [0361] 80.8% of participants claimed that after 8 weeks their skin looked younger by between 5 to 15 years; 50% claimed their skin looked younger by between 10 to 15 years.

[0362] PROAWR2:

[0363] For all three independently assessed metrics (actual wrinkle volume, the appearance of fine lines and wrinkles and the appearance of peri-orbital dark circles): [0364] Both the Test Formulation and the Commercial Product (in-market product A) were significantly better than the control in all three measures—no improvements were seen for the control group. [0365] The Test Formulation was equally effective as the in-market Commercial Product (in-market product A).

[0366] PROAWR1: A home-use study demonstrating the subjective and objective improvements in wrinkles and skin appearance after treatment with a formulation in accordance with the present invention in healthy subjects with ageing and thinning skin

SUMMARY

[0367] This trial was conducted with two anti-ageing formulations in accordance with Example Formulation 1, “Dilute” and “Concentrated”. In the Concentrated version, the tethered ascorbate and tethered peptides were present at twice the concentration that they were in the Dilute version.

[0368] The aim of this initial study was to examine the effectiveness of including these tethered moieties and whether the concentration of them in the formulations and the regime with which they were applied would affect their objective performance and the subjective perception of them by the participants.

[0369] Three regimes were followed: [0370] Dilute: Applied twice a day all over the face and neck [0371] Concentrated:Applied once a day to targeted lines and wrinkles [0372] Combined: Participants on this regime used both products, as directed above

[0373] The results showed that for the combined treatment: [0374] 100% of all participants on all three regimes had measured reduction in wrinkle volume after the first week. [0375] The average scores both wrinkle volume reduction from baseline measurements and the reduction in appearance of wrinkles from baseline scores were highly significant (p<0.001) from week one. [0376] There was mean reduction in actual wrinkle volume of 17.46% at week 12; the mean Glogau (appearance) score had reduced by 25.86% over the same period. [0377] In the subjective assessments: [0378] 100% reported that their skin felt smoother and healthier and looked and felt plumper after 8 weeks [0379] Over 90% reported that their skin looked smoother and healthier, felt more elastic and that the bags and circles under their eyes had reduced [0380] 80.8% of participants claimed that after 8 weeks their skin looked younger by between 5 to 15 years; 50% claimed their skin looked younger by between 10 to 15 years.

TABLE-US-00020 TABLE 11 Summary Protocol Title: A randomised home-use study in three parallel groups of 30 healthy volunteers, with wrinkles, to assess the efficacy of two anti-ageing products and one anti-ageing regime. Study design: Single-blind, within-subject comparison, whole face design. Test Groups: Group Test product Regime Group 1. DH3943 Used twice a day, morning and evening, in an n = 26 even layer to the face and neck Group 2. DH3942 Apply once a day in the evening to targeted n = 27 lines or wrinkles Group 3. DH3943 Apply DH 3943 twice a day in an even layer to n = 26 and the face and neck DH3942 Once a day apply a small quantity of DH3942 to targeted lines or wrinkles. Duration of 12 week treatment period, with assessments at Baseline, Weeks 1, 2, 4, study: 6, 8 and 12. Study Started: w/c 11, Aug. 2014 Study Ended: w/e 21, Nov. 2014 Number of 79 subjects completed the study up to Week 8. 62 subjects completed subjects: the study including the 4 week extension to week 12. Method: Subjects underwent Profilometry assessments after which they were issued with an instruction sheet, diary cards and the test article on Week 0 (baseline). Subjects returned to the Test Centre on Weeks 1, 2, 4, 6 and 8 for Profilometry assessments. Visual assessments were performed on both ageing and peri-orbital dark circles at baseline and Weeks 1, 2, 4, 6 and 8. The subjects completed SPQ's at Week 4 and Week 8. DIC (Digital Image Capture) was performed on approximately 40% of subjects in each group at baseline, Week 2, 4 and 8). A 4 week extension was authorised for the study in which groups 1 and 2 were instructed to continue use of the product and undergo further profilometery and visual assessments at week 12. 50% of Group 3 were instructed to continue use of the regimen whilst the remaining 50% were instructed to use a bland moisturiser. Following week 12 assessments subjects were compensated for their time and inconvenience and exited from the study. Location: Princeton Consumer Research Ltd. Harbour House 23 Chandlers Quay Maldon CM9 4LF United Kingdom

TABLE-US-00021 TABLE 12 Test Products DH3942 - DH3943 - “Dilute” “Concentrated” Soy Phosphatidylcholine Soy Phosphatidylcholine ButyIhydroxytoIuene Butylhydroxytoluene Sodium metabisulphite Sodium metabisulphite Citric acid Citric acid di-Sodium hydrogen di-Sodium hydrogen phosphate dodecahydrate phosphate dodecahydrate Sodium EDTA Sodium EDTA Methyl-4-hydroxybenzoate Methyl-4-hydroxybenzoate Ethyl-4-hydroxybenzoate Ethyl-4-hydroxybenzoate Benzylalcohol Benzylalcohol Polysorbate 80 Polysorbate 80 Sodium hydroxide Sodium hydroxide Carbopol 974P Carbopol 974P Glycerol Glycerol Ethanol Ethanol Water Water Ascorbyl Palmitate Ascorbyl Palmitate* Palmitoyl Tripeptide-1 Palmitoyl Tripeptide-1* Palmitoyl Tetrapeptide-7 Palmitoyl Tetrapeptide-7* Linalool Linalool *In DH3942, these three components were present at double the concentration that they were in DH3943. DH3942 is the same as that in the exemplary composition set out in Example Formulation 1, above.

[0381] Example Formulation 1, above.

[0382] Results

[0383] Effect on Wrinkle Volume

[0384] 100% of participants experienced reductions in wrinkle volume.

TABLE-US-00022 TABLE 13 Average wrinkle volume at time t, expressed as a percentage of average volume at time 0 (=100%). Standard deviation in brackets. P value is the result of a t test comparison to the Week 0 data. Weeks 1 2 4 6 8 12 DH3943 - 96.9% 93.8% 91.9% 90.5% 89.0% “Dilute” (1.3%) (1.6%) (1.9%) (1.9%) (2.1%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 DH3942 - 96.3% 93.7% 90.8% 88.8% 87.1% “Concentrated” (2.1%) (2.7%) (2.7%) (2.7%) (2.7%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Dilute + 96.5% 92.7% 88.9% 85.2% 82.3% 83% Concentrated (0.9%) (1.1%) (1.5%) (1.7%) (1.9%) (1.9%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001

[0385] Effect on Glogau Score (Appearance of Fine Lines and Wrinkles)

[0386] Glogau scores are assessments made by an independent assessor of the severity of appearance of lines and wrinkles.

TABLE-US-00023 TABLE 14 Average Glogau score at time t, expressed as a percentage of average volume at time 0 (=100%). Standard deviation in brackets. P value is the result of a t test comparison to the Week 0 data. Weeks 1 2 4 6 8 12 DH3943 - 99.2%  100% 99.6% 95.4% 88.8% “Dilute” (3.9%) (0.0%) (2.0%) (7.0%)  (8.9%) p = 0.163 N/A p = 0.163 p = 0.001 p < 0.001 DH3942 - 99.2% 98.8% 96.5% 94.7% 93.1% “Concentrated” (4.3%) (4.3%) (7.3%) (8.9%) (12.0%) p = 0.163 p = 0.081 p = 0.008 p = 0.001 p = 0.002 Dilute + 95.7% 91.8% 87.7% 86.5% 81.5% 73.8% Concentrated (6.3%) (7.7%) (7.6%) (8.6%) (10.0%) (10.4%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001

[0387] Subjective Questionnaire Results

TABLE-US-00024 TABLE 15 % of subjects Agreeing/Strongly agreeing with the statement DH 3942 - Conc DH 3943 - Dil Combined Week Week Week Week Week Week Statement 4 8 4 8 4 8 Felt smoother 23.1% 50.0% 30.3% 81.5% 46.2% 100.0% Looked smoother 26.9% 61.5% 55.6% 70.4% 30.8% 96.2% Skin tone more even 50.0% 92.3% 29.6% 74.1% 15.4% 88.5% Acne scarring reduced 19.2% 84.6% 37.0% 55.6% 11.5% 38.5% Blemishes reduced 26.9% 73.1% 29.6% 100.0% 3.9% 80.8% Complexion improved 30.8% 61.5% 22.2% 100.0% 7.7% 76.9% Complexion more radiant 19.2% 38.5% 29.6% 96.3% 23.1% 88.5% Age spots/hyper pigmentation reduced 26.9% 53.9% 55.6% 55.6% 61.5% 88.5% Felt more firm 38.5% 50.0% 55.6% 92.6% 42.3% 80.8% Looked more firm 19.2% 53.9% 22.2% 40.7% 34.6% 100.0% Felt more plump 30.8% 57.7% 40.7% 100.0% 19.2% 100.0% Looked more plump 19.2% 46.2% 59.3% 92.6% 34.6% 100.0% Complexion looked healthier 42.3% 76.9% 25.9% 92.6% 23.1% 96.2% Skin feels healthier 53.9% 61.5% 74.1% 88.9% 42.3% 100.0 Skin felt more elastic 46.2% 73.1% 55.6% 70.4% 26.9% 92.3% Bags/circles reduced 30.8% 57.7% 44.4% 96.3% 30.8% 92.3% Most effective anti-ageing used 42.3% 76.9% 55.6% 81.5% 30.8% 76.9% Would use instead of current 46.2% 69.2% 55.6% 92.6% 65.4% 92.3% Easy to use alongside current products 46.2% 80.8% 100.0% 96.3% 34.6% 61.5% Would recommend to a friend 65.4% 76.9% 81.5% 100.0% 73.1% 100.0% Would buy the product 38.5% 76.9% 18.5% 81.5% 84.6% 92.3% Skin looked younger by 20+ years 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 15+ years 0.0% 3.9% 0.0% 3.7% 0.0% 7.7% 10+ years 3.9% 19.2% 7.4% 22.2% 3.9% 42.3% 5+ years 23.1% 42.3% 81.5% 55.6% 42.3% 30.8% Same 73.1% 34.6% 11.1% 18.5% 50.0% 19.2%

[0388] PROAWR2: A randomised home-use study in three parallel groups of 30 healthy volunteers, with wrinkles, to assess the efficacy of one anti-ageing product against a placebo and a comparator with an additional 73 subjects to complete a subjective user-trial using only the product

SUMMARY

[0389] This trial tested a preferred test anti-ageing formulation in accordance with the present invention (the “Concentrated” formulas from PROAWR1), against an in-market anti-ageing Commercial Product (in-market product A) and a control formulation.

[0390] The aim of this study was to test: [0391] a) That previous results seen with the preferred formulation were real (control comparison) [0392] b) To assess the effectiveness against a currently marketed formulation for anti-ageing.

[0393] The results showed that:

[0394] For all three independently assessed metrics (actual wrinkle volume, the appearance of fine lines and wrinkles and the appearance of peri-orbital dark circles): [0395] Both the Test Formulation and the Commercial Product were significantly better than the control in all three measures—no improvements were seen for the control group. [0396] The Test Formulation was equally effective as the in-market Commercial Product.

[0397] For the subjective assessment of the products: [0398] Both the Test Formulation and the Commercial Product were significantly better than the control in all measures [0399] The Test Formulation was rated higher than the Commercial Product for: [0400] Skin looked smoother [0401] Skin felt more plump [0402] Skin felt healthier [0403] Skin felt more elastic [0404] Complexion looked healthier [0405] Skin tone looked more even [0406] Eye puffiness reduction [0407] Effectiveness in anti-ageing [0408] Lifted sagging skin [0409] Hydration of skin [0410] Reducing pore size

TABLE-US-00025 TABLE 16 Summary Protocol Title: A randomised home-use study in three parallel groups of 30 healthy volunteers, with wrinkles, to assess the efficacy of one anti-ageing product against a placebo and a comparator with an additional 70 subjects to complete a subjective user-trial using only the product. Study design: Single-blind, within-subject comparison, whole face design. Test Groups: Group Test product Regime Group 1. Test Formulation Apply twice a day Group 2. Control Apply all over to Group 3. In-market product A face and neck Apply additional amounts to areas where wrinkles/ageing is more noticeable Apply a thin layer to the sub-orbital eye sacks Duration of 8 week treatment period, with assessments at Baseline, Week 4, and study: Week 8. Study Started: w/c 16, Feb. 2015 Study Ended: w/e 24, Apr. 2015 Method: Subjects underwent Profilometry assessments after which they were issued with an instruction sheet, diary cards and the test article on Week 0 (baseline). Subjects returned to the Test Centre on Weeks 1, 2, 4, 6 and 8 for Profilometry assessments. When the subjects returned to the Test Centre on Week 8, they returned any unused test articles and their completed diary cards. Visual assessments were performed on both ageing and peri-orbital dark circles at baseline and Weeks 1, 2, 4, 6 and 8. The subjects completed SPQ's at Week 8. Professional photography was performed on approximately 40% of subjects in each group at baseline, and Weeks 1, 2, 4 and 8 with as many subjects as possible for group 1 being photographed above and beyond 40%. 70 subjects were also issued the product to use at home for the duration of the test period and completed an SPQ at the same time points as the active groups. Location: Princeton Consumer Research Ltd. Harbour House 23 Chandlers Quay Maldon CM9 4LF United Kingdom

TABLE-US-00026 TABLE 17 Test Products Test Formulation Control Soy Phosphatidylcholine ButyIhydroxytoIuene Ethanol Methyl-4-hydroxybenzoate Methyl-4-hydroxybenzoate Ethyl-4-hydroxybenzoate Ethyl-4-hydroxybenzoate Benzylalcohol Polysorbate 80 Linalool Ascorbyl Palmitate Palmitoyl Tripeptide-1 Palmitoyl Tetrapeptide-7 Sodium metabisulphite Citric acid monohydrate Citric acid monohydrate di-Sodium hydrogen di-Sodium hydrogen orthophosphate anhydrous orthophosphate anhydrous Sodium EDTA Sodium hydroxide Sodium hydroxide Carbopol 974P Carbopol 974P Glycerol Water Water

[0411] Results

[0412] Effect on Wrinkle Volume

TABLE-US-00027 TABLE 18 Average wrinkle volume at time t, expressed as a percentage of average volume at time 0 (= 100%). Standard deviation in brackets. P value is the result of a t test comparison to the Week 0 data. n Week 1 Week 2 Week 4 Week 6 Week 8 Test 27 95.9% 93.1% 90.7% 88.8% 87.5% Formulation (2.0%) (2.9%) (3.6%) (3.9%) (4.1%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Control 28 100.2%  100.5%  100.3%  100.4%  100.5%  (1.0%) (2.0%) (2.9%) (3.0%) (3.3%) P = 0.31  P = 0.17  P = 0.57  P = 0.51  P = 0.39  In-market 29 95.8% 92.9% 90.5% 88.1% 86.2% product A (2.5%) (3.3%) (3.8%) (4.1%) (4.3%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001

TABLE-US-00028 TABLE 19 p values for t-tests between different treatments Week 1 Week 2 Week 4 Week 6 Week 8 Test Formulation vs. p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Control Test Formulation vs. in- p = 0.554 p = 0.623 p = 0.604 p = 0.817 p = 0.947 market product A In-market product A vs. p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Control [0413] Test Formulation and in-market product A significantly different from Control. [0414] No significant difference between Test Formulation and in-market product A.

[0415] Effect on Glogau Score (Appearance of Fine Lines and Wrinkles)

[0416] Glogau scores are assessments made by an independent assessor of the severity of appearance of lines and wrinkles.

TABLE-US-00029 TABLE 20 Average Glogau score at time t, expressed as a percentage of average volume at time 0 (=100%). Standard deviation in brackets. P value is the result of a t test comparison to the Week 0 data. n Week 1 Week 2 Week 4 Week 6 Week 8 Test 27 92.6% 90.7% 79.5% 78.5% 73.9% Formulation (8.5%) (8.3%) (9.8%) (8.9%)  (7.4%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Control 28 101.7%  100.0%  102.8%  105.0%  103.9%  (10.8%)  (10.7%)  (8.0%) (13.4%)  (12.8%) p = 0.81  p = 0.79  p = 0.21  p = 0.12  p = 0.31  In-market 29 91.6% 88.9% 82.2% 79.9% 74.3% product A (8.1%) (8.4%) (8.5%) (9.5%) (10.5%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001

TABLE-US-00030 TABLE 21 p values for t-tests between different treatments Week 1 Week 2 Week 4 Week 6 Week 8 Test Formulation vs. p = 0.266 p = 0.198 p = 0.004 p < 0.001 p < 0.001 Control Test Formulation vs. in- p = 0.976 p = 0.998 p = 0.681 p = 0.698 p = 0.799 market product A in-market product A vs. p = 0.318 p = 0.245 p = 0.018 p = 0.005 p = 0.001 Control [0417] Test Formulation and in-market product A significantly different from Control from week 4. [0418] No significant difference between Test Formulation and in-market product A.

[0419] Effect on Peri-Orbital Dark Circles

TABLE-US-00031 TABLE 22 Average PO Dark circle score at time t, expressed as a percentage of average volume at time 0 (=100%). Standard deviation in brackets. P value is the result of a t test comparison to the Week 0 data. n Week 1 Week 2 Week 4 Week 6 Week 8 Test 27 72.2% 48.1% 38.9% 29.6% 27.8% Formulation (34.9%) (37.9%) (42.4%) (42.2%) (37.6%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Control 28  100% 112.5%  114.3%  119.6%  121.4%    (0%) (44.4%) (54.2%) (59.8%) (58.4%) p = N/A p = 0.49  p = 0.79  p = 0.63  p = 0.45  In-market 29 77.6% 56.9% 34.5% 34.5% 31.0% product A (34.3%) (43.8%) (40.3%)  42.5%) (41.0%) p < 0.001 p < 0.001 p < 0.001 p < 0.001 p < 0.001

TABLE-US-00032 TABLE 23 p values for t-tests between different treatments Week 1 Week 2 Week 4 Week 6 Week 8 Test Formulation vs. p = 0.013 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Control Test Formulation vs. in- p = 0.949 p = 0.589 p = 0.794 p = 0.562 p = 0.962 market product A in-market product A vs. p = 0.008 p < 0.001 p < 0.001 p < 0.001 p < 0.001 Control [0420] Test Formulation and in-market product A significantly different from Control from week 1. [0421] No significant difference between Test Formulation and in-market product A.

[0422] Subjective Questionnaire Results

TABLE-US-00033 TABLE 24 In market Test Product Formulation A Control N = 27 + 73* N = 26 N = 28 After using the product. . . my skin felt significantly smoother 82.00% 89.66% 28.57% my skin looked significantly smoother 85.00% 55.17% 46.43% my skin felt significantly more firm 89.00% 89.66% 42.86% my skin looked significantly more firm 54.00% 58.62% 46.43% my skin felt significantly more plump 86.21% 25.00% my skin looked significantly more plump 56.00% 68.97% 25.00% my skin feels significantly healthier 86.00% 79.31% 46.43% my skin felt more elastic 83.00% 82.76% 28.57% my complexion was visibly improved 58.00% 65.52% 25.00% my complexion was visibly more radiant 61.00% 82.76% 21.43% my complexion looked significantly healthier 90.00% 86.21% 32.14% my skin tone looked significantly more even 90.00% 79.31% 25.00% any age spots/hyperpigmentation were 67.00% 72.41% 46.43% significantly reduced the bags under my eyes were significantly 89.00% 35.71% reduced the puffiness all around my eyes was 88.00% 86.21% 39.29% significantly reduced the dark circles under my eyes were 86.00% 89.66% 42.86% significantly reduced sagging skin was lifted 89.00% 86.21% 46.43% my skin looked: at least 5 years younger 76.00% 82.76% 39.29% at least 10 years younger 35.00% 34.48%  0.00% The product. . . redefined sagged skin 58.00% 75.86% 28.57% has a pleasant fragrance 62.00% 58.62% 42.86% has a pleasant consistency and texture 79.31% 32.14% has a pleasant appearance 61.00% 65.52% 28.57% felt good on my skin after or upon 66.00% 65.52% 32.14% application dried quickly 89.00% 32.14% was easy to use alongside my usual 86.21% 28.57% skincare products was the most effective anti-ageing serum I 86.00% 62.07% 39.29% have used redefined facial contours that were sagged 67.00% 72.41% 39.29% hydrated my skin 86.21% 25.00% has visibly reduced the size of my pores 85.00% 68.97% 17.86% has visibly reduced the appearance of any 69.00% 75.86% 25.00% thread veins I would. . . recommend the product to a friend 79.00% 32.14% buy this product in place of my usual anti- 57.14% ageing serum *An additional 73 subjects used the Test Formulation and answered the self-assessment questionnaire but were not independently assessed for wrinkle volume, wrinkle appearance or appearance of peri-orbital dark circles.

Example 5: A Study in Volunteers to Investigate the Sensation Produced Using a Formulation Comprising Capsaicin

BACKGROUND

[0423] There are several products on the market that provide a warming sensation when applied to the joint or muscle, to relieve aches or pains. These products contain chemicals such as capsaicin which stimulate the nerve endings in the skin to create the warming sensations (without actually being related to a physical temperature change).

[0424] The inventors have developed a capsaicin-containing formulation in accordance with the present invention, so that a warming sensation is associated with the product, either at application or sometime after. The formulation provides a combined effect of the joint-pain relieving property of drug-free colloidal dispersions, combined with a warming feeling.

[0425] The formulation (TP2) comprised insoluble and inflexible liposomes comprising capsaicin and menthol in combination with Transfersomes™ comprising menthol.

[0426] Transfersomes™/Sequessomes™/Tethersomes are flexible, deformable colloidal particles that make their own way into the skin, whereas liposomes are rigid vesicles that would not ordinarily be expected to penetrate the skin by themselves unless some other force is applied (e.g. Sequessomes™/Transfersomes™/Tethersomes in the same mixture opening pores and pulling/pushing the liposomes through).

[0427] The starting material used to make TP2 is set out below.

[0428] TP2 (PD-14-0073) was formed by taking 90 g of Start Material 1 and adding/mixing in 10 g of Start Material 2.

[0429] Start Material 1 (Flexible, Empty, Menthol-Fragranced Vesicles):

TABLE-US-00034 Quantity Required Per 100 g Final Product (g) Organic Phase - Empty Transfersome Intermediate SPC (Dry Mass) 6.87000 Ethanol 3.65100 BHT 0.02000 Methyl-4-hydroxybenzoate 0.25000 Ethyl-4-hydroxybenzoate 0.25000 Benzyl alcohol 0.52500 Polysorbate 80 0.85000 Capsaicin 0.00000 Menthol 0.10000 Organic Phase Sub-Total 12.51600 Aqueous Phase - Empty Transfersome Intermediate Sodium dihydrogen orthophosphate 2 H.sub.2O 0.03700 disodium hydrogen orthophosphate 12 H.sub.2O 0.45300 Sodium EDTA 0.10000 Water 45.22700 Aqueous Phase Sub-Total 45.81700 Intermediate Transfersome Total 58.33300 Gel Phase - Final Product Sodium dihydrogen orthophosphate 2 H.sub.2O 0.02400 disodium hydrogen orthophosphate 12 H.sub.2O 0.30200 Sodium hydroxide 0.63000 Carbopol 974P NF 1.25000 Glycerol 3.00000 Water 36.46100 Gel Phase Sub-Total 41.66700 Overall Total 100.00000

[0430] Start Material 2 (Capsaisin Liposomes):

TABLE-US-00035 Quantity Required Per 100 a Final Product (g) Organic Phase - Liposome Intermediate SPC (Dry Mass) 6.87000 Ethanol 4.25100 BHT 0.02000 Methyl-4-hydroxybenzoate 0.25000 Ethyl-4-hydroxybenzoate 0.25000 Benzyl alcohol 0.52500 Polysorbate 80 0.00000 Capsaicin 0.25000 Menthol 0.10000 Organic Phase Sub-Total 12.51600 Aqueous Phase - Liposome Intermediate Sodium dihydrogen orthophosphate 2 H.sub.2O 0.03700 disodium hydrogen orthophosphate 12 H.sub.2O 0.45300 Sodium EDTA 0.10000 Water 45.22700 Aqueous Phase Sub-Total 45.81700 Liposome Intermediate Total 58.33300 Gel Phase - Final Product Sodium dihydrogen orthophosphate 2 H.sub.2O 0.02400 disodium hydrogen orthophosphate 12 H.sub.2O 0.30200 Sodium hydroxide 0.63000 Carbopol 974P NF 1.25000 Glycerol 3.00000 Water 36.46100 Gel Phase Sub-Total 41.66700 Overall Total 100.00000

[0431] Summary Formulae:

TABLE-US-00036 Title PD-14-0073 (90% Start Material 1 plus 10% Start Ingredient Start Material 1 Start Material 2 Material 2 SPC (dry mass) 6.870 g 6.870 g 6.870 g Polysorbate 80 0.850 g — 0.765 g Benzylalcohol 0.525 g 0.525 g 0.525 g Methyl-4-hydroxybenzoate 0.250 g 0.250 g 0.250 g Ethyl-4-hydroxybenzoate 0.250 g 0.250 g 0.250 g Butylhydroxytoluene 0.020 g 0.020 g 0.020 g Disodium EDTA 0.100 g 0.100 g 0.100 g Disodium hydrogen phosphate 0.755 g 0.755 g 0.755 g 12 H.sub.2O Sodium dihydrogen phosphate 2 0.061 g 0.061 g 0.061 g H.sub.2O Glycerol 3.000 g 3.000 g 3.000 g Ethanol 3.651 g 4.251 g 3.711 g Sodium hydroxide 0.630 g 0.630 g 0.630 g Carbopol 974P NF 1.250 g 1.250 g 1.250 g Water 81.688 g 81.688 g 81.688 g Agent of interest - in continuous phase Capsaicin — 0.250 g 0.025 g Agent of interest - in Transfersome Menthol 0.100 g 0.100 g 0.100 g Total mass 100.000 g 100.000 g 100.000 g

[0432] Results

[0433] The formulation was tried by three individuals. The results are presented in the table below.

TABLE-US-00037 TABLE 26 Test product TP2 (PD-14-0073) Capsaicin In liposomes Menthol In membranes of Transfersomes ™and of liposomes CAPSAICIN: Onset of sensation 5 to 30 mins Onset of optimal sensation 15 to 60 mins Duration of sensation 45 to 180 mins Intensity of sensation Pleasant MENTHOL: Duration of sensation 15 to 30 mins Intensity of sensation Enough

[0434] Conclusion/Summary

[0435] These early results are useful in showing that: [0436] The warming sensation of capsaicin can be felt: [0437] In Transfersomes™—in the same “phase” as a joint-targeting colloidal particle [0438] In liposomes—separate “phase” to the joint-targeting colloidal particles

Example 6: A User Trial Study in Healthy Volunteers to Investigate the Efficacy of a Formulation Comprising Caffeine in Improving the Appearance of Periorbital Skin

[0439] This study tested the efficacy of a formulation comprising caffeine and tocopherol in the continuous phase and three types of deformable colloidal particles: 1) Tethersomes to which palmitoyl ascorbate is tethered, 2) Tethersomes to which palmitoyl tripeptide-1 is tethered, and 3) Tethersomes to which palmitoyl tetrapeptide-7 is tethered.

[0440] Methods

[0441] Summary Protocol

TABLE-US-00038 Study desiqn: Sinqle-blind Test article: CBL-DERM-15-013 Periorbital skin serum Duration of treatment: 4 weeks Number of subjects: 102 Type of subjects: Healthy male and female subjects (from a breadth of ethnicities) aged between 40 and 70 years old (in equal proportions, 33% each 10 year age group), from a breadth of ethnicities and with a variety of skin types (normal, combination, dry, oily, sensitive) who suffer from the appearance of two of the following characteristics: Periorbital puffiness - swelling around the eyes Puffiness below the lower eyelids - ‘eye bags’ Periorbital dark circles caused by a) aging b) heredity and c) hyperpigmentation (dark skinned people) (There must be an equal representation of all three characteristics to ensure claims objectivity). All subjects must have visible wrinkles, fine lines and crow's feet around the eye area Observations: Subjects were asked to apply the product as per usage instructions. They were then asked to complete a Self- Perception Questionnaire (SPQ) after two weeks and at the end of the study after four weeks. Treatment: Subjects were issued with samples of the test article and directions for use following standard in-use application regime. Location Princeton Consumer Research Harbour House 23 Chandlers Quay Maldon CM9 4LF United Kingdom

[0442] Formulation (CBL-DERM-15-013)

TABLE-US-00039 Quantity Required Per 100 g Final Product (g) Organic Phase - 1600 ppm PAA Intermediate Soy phosphatidylcholine (SPC) 4.12200 Ethanol 1.99660 Butyl hydroxyanisole (BHA) 0.01200 Methyl-4-hydroxybenzoate 0.15000 Ethyl-4-hydroxybenzoate 0.15000 Benzyl alcohol 0.31500 Polysorbate 80 0.40800 Palmitoyl ascorbic acid 0.09600 +/− alpha tocopherol 0.20000 Linalool 0.06000 Organic Phase Sub-Total 7.50960 Aqueous Phase - 1600 ppm PAA intermediate Sodium metabisulphite 0.03000 Citric acid monohydrate 0.04320 Disodium EDTA 0.06000 Disodium hydrogen orthophosphate 0.17280 dodecahydrate Caffeine 0.05000 Water 27.13420 Aqueous Phase Sub-Total 27.49020 PAA Tethersome Intermediate Total 34.99980 Organic Phase - 300 ppm Tetrapeptide Intermediate Soy phosphatidylcholine (SPC) 1.37400 Ethanol 0.74120 Butylhydroxyanisole (BHA) 0.00400 Methyl-4-hydroxybenzoate 0.05000 Ethyl-4-hydroxybenzoate 0.05000 Benzyl alcohol 0.10500 Polysorbate 80 0.15300 Palmitoyl peptide 0.00600 Linalool 0.02000 Organic Phase Sub-Total 2.50320 Aqueous Phase - 300 ppm Tetrapeptide Intermediate Sodium metabisulphite 0.01000 Citric acid monohydrate 0.01440 Disodium EDTA 0.02000 Disodium hydrogen orthophosphate 0.05760 dodecahydrate Water 9.06140 Aqueous Phase Sub-Total 9.16340 Tetrapeptide Tethersome 11.66660 Intermediate Total Organic Phase - 300 ppm Tripeptide Intermediate Soy phosphatidylcholine (SPC) 1.37400 Ethanol 0.74120 Butylhydroxyanisole (BHA) 0.00400 Methyl-4-hydroxybenzoate 0.05000 Ethyl-4-hydroxybenzoate 0.05000 Benzyl alcohol 0.10500 Polysorbate 80 0.15300 Palmitoyl peptide 0.00600 Linalool 0.02000 Organic Phase Sub-Total 2.50320 Aqueous Phase - 300 ppm Tripeptide Intermediate Sodium metabisulphite 0.01000 Citric acid monohydrate 0.01440 Disodium EDTA 0.02000 Disodium hydrogen orthophosphate 0.05760 dodecahydrate Water 9.06140 Aqueous Phase Sub-Total 9.16340 Tripeptide Tethersome 11.66660 Intermediate Total Sum of Sequessome and 58.33300 Tethersome Intermediates Gel Phase - Final Product Sodium hydroxide 0.16000 Carbopol 974P NF 0.75000 Glycerol 3.00000 Citric acid monohydrate 0.05600 Disodium hydrogen orthophosphate 0.24200 dodecahydrate Water 37.45900 Gel Phase Sub-Total 41.66700 Overall Total 100.00000

[0443] Summary Formulae of CBL-DERM-15-013

TABLE-US-00040 Periorbital Ingredient (percentage; g per 100 g) Skin SPC (dry mass) 6.870 Polysorbate 80 0.714 Benzylalcohol 0.525 Methyl-4-hydroxybenzoate 0.250 Ethyl-4-hydroxybenzoate 0.250 Butylhydroxyanisole 0.020 Linalool 0.100 Sodium metabisulphite 0.050 Disodium EDTA 0.100 Disodium hydrogen phosphate 12 H.sub.2O 0.530 Citric acid monohydrate 0.128 Glycerol 3.000 Ethanol *** 3.479 Sodium hydroxide 0.160 Carbopol 974P NF 0.750 Water 82.716  Agent of interest - tethered to Tethersomes Palmitoyl tripeptide-1 0.006 Palmitoyl tetrapeptide-7 0.006 Palmitoyl ascorbate 0.096 Agent of interest - in continuous phase Tocopherol 0.200 Caffeine 0.050 pH 5.5*  *Final pH approximate; to be confirmed

[0444] Results

[0445] Within-treatment analysis (p<0.05) of periorbital clinical assessment shows there to be a statistically significant reduction in wrinkles (15.69%), fine lines (31.22%), crow's feet (21.47%), puffiness (38.07%), dark circles (26.01%) and eye bags (38.94%) after 4 weeks of product usage.

[0446] The product performed statistically favourably over the 4 week study in the majority of attributes under Clearcast guidelines of advertising. The product showed a preference or favourability in the majority of attributes: [0447] After using the product, 96.08% of users agreed, or strongly agreed the product reduced the appearance of puffiness and swelling around their eyes. [0448] After using the product, 96.08% of users agreed, or strongly agreed their skin felt and looked less thin. [0449] After using the product, 93.14% of users agreed, or strongly agreed their skin felt more elastic. [0450] After using the product, 96.08% of users agreed, or strongly agreed the product reduced the appearance of swelling and puffiness below the lower eyelids (eye bags). [0451] After using the product, 90.20% of users agreed, or strongly agreed the product lifted sagged skin (reduction in skin droopiness and sagging). [0452] After using the product, 92.16% of users agreed, or strongly agreed the product reduced the appearance of under eye dark circles. [0453] After using the product, 95.10% of users agreed, or strongly agreed their skin looked and felt more firm. [0454] After using the product, 90.20% of users agreed, or strongly agreed there was a reduction in fine lines (inc. crow's feet) around the eye area. [0455] After using the product, 87.25% of users agreed, or strongly agreed there was a reduction in deep wrinkles (inc. crow's feet) around the eye area. [0456] After using the product, 93.14% of users agreed, or strongly agreed their skin tone looked more even. [0457] After using the product, 90.20% of users agreed, or strongly agreed their eye contor looked and felt tighter. [0458] After using the product, 90.20% of users agreed, or strongly agreed their eye contour looked more toned/lifted. [0459] After using the product, 96.08% of users agreed, or strongly agreed their eyes looked rested/less tired. [0460] After using the product, 97.06% of users agreed, or strongly agreed their eye skin was more hydrated. [0461] After using the product, 95.10% of users agreed, or strongly agreed their skin looked smoother. [0462] After using the product, 99.02% of users agreed, or strongly agreed the product was gentle and well tolerated. [0463] After using the product, 75.49% of users stated, yes, they would buy this product instead of their usual product. [0464] After using the product, 86.27% of users stated, yes, they would recommend this product to a friend. [0465] After using the product, 46.08% of users stated, they looked at least 5 years younger.

Example 7: A User Trial Study in Healthy Volunteers to Investigate the Efficacy of a Formulation Comprising Salicylate and Tocopherol in Improving Skin Tone

[0466] This study tested the efficacy of a formulation comprising tocopherol in the continuous phase and two types of colloidal particles: 1) Tethersomes to which tridecyl salicylate is tethered and 2) Tethersomes to which palmitoyl ascorbate and tocopheryl linoleate are tethered.

[0467] Methods

[0468] Summary Protocol

TABLE-US-00041 Study design: Single-blind Test article: CBL-DERM-15-014 skin tone serum Duration of treatment: 6 weeks Number of subjects: 110 Type of subjects: A user trial study in 110 healthy male (20%) and female (80%) subjects aged between 20 and 70 years old (equal proportions of each 10 year age group; 20's, 30's, 40's, 50's, 60's, 70's); from a breadth of ethnicities with a variety of skin types who suffer from one pronounced or two out of the following six conditions; melasma or chloasma spots, uneven skin tone, age, sun or “liver” spots, freckles, post-inflammatory hyperpigmentation or periorbital hyperpigmentation (dark circles). Observations: Subjects were asked to apply the product as per usage instructions. They were asked to complete a Self-Perception Questionnaire (SPQ) at the end of Weeks 3 and 6. Treatment: Subjects were issued with samples of the test article and directions for use following standard in-use application regime. Location Princeton Consumer Research Harbour House 23 Chandlers Quay Maldon CM9 4LF United Kingdom

[0469] Formulation (CBL-DERM-15-014)

TABLE-US-00042 Quantity Required Per 100 g Final Product (g) Organic Phase - Tridecyl Salicylate Intermediate Soy phosphatidylcholine (SPC) 3.43500 Ethanol 1.67550 Butylhydroxyanisole (BHA) 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.42500 Tridecyl Salicylate 0.10000 +/− alpha tocopherol 0.05000 Linalool 0.05000 Organic Phase Sub-Total 6.25800 Aqueous Phase - Tridecyl Salicylate Intermediate Citric acid monohydrate 0.03600 Disodium EDTA 0.05000 Sodium metabisulphite 0.02500 Disodium hydrogen orthophosphate 0.14400 dodecahydrate Water 22.65350 Aqueous Phase Sub-Total 22.90850 Tridecyl Salicylate Tethersome 29.16650 Intermediate Total Organic Phase - PAA/Tocopheryl Linoleate Intermediate Soy phosphatidylcholine (SPC) 3.43500 Ethanol 1.57950 Butylhydroxyanisole (BHA) 0.01000 Methyl-4-hydroxybenzoate 0.12500 Ethyl-4-hydroxybenzoate 0.12500 Benzyl alcohol 0.26250 Polysorbate 80 0.42500 Palmitoyl ascorbic acid 0.09600 Tocopheryl linoleate 0.10000 +/− alpha tocopherol 0.05000 Linalool 0.05000 Organic Phase Sub-Total 6.25800 Aqueous Phase - PAA/Tocopheryl Linoleate Intermediate Citric acid monohydrate 0.03600 Disodium EDTA 0.05000 Sodium metabisulphite 0.02500 Disodium hydrogen orthophosphate 0.14400 dodecahydrate Water 22.65350 Aqueous Phase Sub-Total 22.90850 PAA/Tocopheryl Linoleate Tethersome 29.16650 Intermediate Total Sum of Sequessome and 58.33300 Tethersome Intermediates Gel Phase - Final Product Sodium hydroxide 0.16000 Carbopol 974P NF 0.75000 Glycerol 3.00000 Citric acid monohydrate 0.05600 Disodium hydrogen orthophosphate 0.24200 dodecahydrate Water 37.45900 Gel Phase Sub-Total 41.66700 Overall Total 100.00000

[0470] Summary Formulae of CBL-DERM-15-014

TABLE-US-00043 Ingredient (percentage; g per 100 g) Uneven Skin Tone ** SPC (dry mass) 6.870 Polysorbate 80 0.850 Benzylalcohol 0.525 Methyl-4-hydroxybenzoate 0.250 Ethyl-4-hydroxybenzoate 0.250 Butylhydroxyanisole 0.020 Linalool 0.100 Sodium metabisulphite 0.050 Disodium EDTA 0.100 Disodium hydrogen phosphate 12 H.sub.2O 0.530 Citric acid monohydrate 0.128 Glycerol 3.000 Ethanol *** 3.255 Sodium hydroxide 0.160 Carbopol 974P NF 0.750 Water 82.766  Agent of interest - tethered to Tethersomes Palmitoyl ascorbate 0.096 Tocopheryl Linoleate 0.100 Tridecyl Salicylate 0.100 Agent of interest - in continuous phase Tocopherol 0.100 pH 5.5 *  * Final pH approximate; to be confirmed ** The source of SC dry mass for Uneven Skin Tone will be lipoid Phospholipon 90K (P90K).

[0471] The quantity of P90K and ethanol to be added should be calculated accordingly.

[0472] Results

[0473] The product performed statistically favourably over the 6 week study in the majority of attributes under Clearcast guidelines of advertising. The product did show a preference or favourability in the majority of attributes: [0474] After using the product, 100% of users agreed, or strongly agreed the product reduced the appearance of fine lines and wrinkles. [0475] After using the product, 100% of users agreed, or strongly agreed their skin looked significantly smoother. [0476] After using the product, 100% of users agreed, or strongly agreed their skin felt significantly more healthy. [0477] After using the product, 96.36% of users agreed, or strongly agreed their skin felt more elastic. [0478] After using the product, 99.09% of users agreed, or strongly agreed their complexion looks younger. [0479] After using the product, 99.09% of users agreed, or strongly agreed their complexion is visibly improved. [0480] After using the product, 100% of users agreed, or strongly agreed their complexion was visibly more radiant. [0481] After using the product, 100% of users agreed, or strongly agreed their complexion looked significantly healthier. [0482] After using the product, 98.18% of users agreed, or strongly agreed their skin tone looked significantly more even. [0483] After using the product, 96.36% of users agreed, or strongly agreed that the amount of hyperpigmentation/pigmented skin blemishes were significantly reduced. [0484] After using the product, 97.27% of users agreed, or strongly agreed that the size of hyperpigmentation/pigmented skin blemishes were significantly reduced. [0485] After using the product, 99.09% of users agreed, or strongly agreed that hyperpigmentation/pigmented skin blemishes were significantly lighter. [0486] After using the product, 20.00% of users agreed, or strongly agreed that hyperpigmented/pigmented skin blemishes totally disappeared. [0487] After using the product, 100% of users agreed, or strongly agreed their periorbital dark circles got significantly lighter. [0488] After using the product, 95.45% of users agreed, or strongly agreed this was the most effective hyperpigmentation solution they had used. [0489] After using the product, 92.73% of users stated, yes, they would buy this product instead of their usual product. [0490] After using the product, 100% of users stated, yes, they would recommend this product to a friend.

[0491] With reference to packaging/promotional claims, any results with a top 3 & 4 scoring greater than 80% are highly favourable.