Composition suitable to preserve the physiological condition of skin and hair and reestablish their regenerative functions

Abstract

The present invention relates to a composition for pharmaceutical, nutritional or cosmetic use, suitable to preserve the physiological condition and health of skin and hair and to reestablish their regenerative functions, characterized in that it comprises a mixture of carnitine, caffeine and arginine as active principle, as such or as derivatives, such as pharmacologically acceptable salts. The effect of preserving the physiologic condition and health of skin and hair, and of reestablishing their regenerative functions is mainly achieved through an increase in ATP production by skin and hair cells.

Claims

1. A mixture of carnitine, caffeine and arginine, or pharmacologically acceptable salts or simple esters thereof, as active principle in a nutritional or cosmetic composition for preserving the physiological condition and health of skin or hair and for reestablishing their regenerative functions, wherein the ratio by weight of carnitine, caffeine, and arginine in the mixture is 1:1:3, respectively.

2. The mixture according to claim 1, wherein the mixture is made of carnitine, caffeine and arginine as active principle according to the following concentrations: carnitine 10 mg/ml, caffeine 10 mg/ml, arginine 30 mg/ml.

3. The mixture according to claim 1 for topical administration, wherein the mixture is made of carnitine, caffeine, arginine as active principle according to the following concentrations: carnitine from 1.0 to 33.3 mg/ml, caffeine from 1.0 to 33.3 mg/ml, arginine from 3 to 100 mg/ml.

4. The mixture according to claim 3, wherein the mixture is made of carnitine, caffeine and arginine as active principle according to the following concentrations: carnitine 2-10 mg/ml, caffeine 2-10 mg/ml, arginine 6-30 mg/ml.

5. The mixture according to claim 1 for oral and systemic administration, wherein the mixture is made of carnitine, caffeine, arginine as active principle according to a daily administration dose within the following ranges: carnitine 4-400 mg, caffeine 4-400 mg, arginine 12-1200 mg.

6. The mixture according to claim 5, wherein the mixture is made of carnitine, caffeine, arginine as active principle according to a daily administration dose within the following ranges: carnitine 10-100 mg, caffeine 10-100 mg, arginine 30-300 mg.

7. A method for preserving the physiological condition and health of skin or hair, or for reestablishing their regenerative functions in a subject in need thereof, said method comprising: administering, to the subject, a mixture of carnitine, caffeine and arginine, or pharmacologically acceptable salts or simple esters thereof, wherein the ratio by weight of carnitine, caffeine, and arginine in the mixture is 1:1:3, respectively, and wherein said administering is carried out at a dose effective to preserve the physiological condition and health of the subject's skin or hair or reestablish their regenerative functions in said subject.

8. The method according to claim 7, wherein said administering is carried out at a dose effective to preserve the physiological condition and health of the subject's skin.

9. The method according to claim 7, wherein said administering is carried out at a dose effective to preserve the physiological condition and health of the subject's hair.

10. The method according to claim 9, wherein said administering is carried out at a dose effective to contrast hair loss in the subject.

11. The method according to claim 9, wherein said administering is carried out at a dose effective to promote hair regrowth in the subject.

12. The method according to claim 7, wherein the mixture is made of carnitine, caffeine, arginine as active principle according to the following concentrations: carnitine from 1.0 to 33.3 mg/ml, caffeine from 1.0 to 33.3 mg/ml, arginine from 3 to 100 mg/ml, and it is administered topically.

13. The method according to claim 7, wherein the mixture is made of carnitine, caffeine, arginine as active principle according to a daily administration dose within the following ranges: carnitine 4-400 mg, caffeine 4-400 mg, arginine 12-1200 mg, and it is administered orally.

Description

DRAWINGS

(1) THE diagrams according to the figures in the accompanying drawings show the results for the experimental study described hereafter.

(2) FIG. 1 shows a diagram for the production of cellular ATP for the mixture of the invention compared with other reference products.

(3) FIG. 2 shows a diagram of the corresponding increase % in the production of cellular ATP.

EXPERIMENTAL STUDY: ATP ASSAY

(4) Materials and Methods

(5) Cells

(6) Human Hair Follicle Outer Root Sheath Cells (HHFORSC), supplied by Innoprot, are isolated by ScienCell Research Labs from the outer sheath of the root of human hair.

(7) The cell line was grown in culture medium for mesenchymal stem cells (MSCM): 500 mL basal medium, 20% fetal bovine serum (FBS), 1% growth supplement for mesenchymal stem cells (MSCGS), 1% penicillin/streptomycin (P/S solution) and maintained in 25 cm.sup.2 culture flasks at 37 C. and 5% CO.sub.2.

(8) Before proceeding with plating the cells, the flask cells is coated with poly-L-lysine (2 g/cm.sup.2).

(9) Every two days, the confluent cultures are divided 1:3-1:6, after washing with DPBS (Dulbecco's Phosphate-Buffered Saline), using a solution of T/E (trypsin/EDTA solution) and TNS (Trypsin Neutralization Solution) and seeded at 2-5.sup.4 cell/cm.sup.2, 37 C., 5% CO.sub.2.

(10) Treatments and ATP Assay

(11) Experimental Procedure

(12) Day 1: Seed Cells

(13) When the cells (HHFORSC) reach about 80% confluence, they are detached with trypsin/EDTA and seeded at a density of 110.sup.6 cells/ml in 12-well plates and then incubated at 37 C., 5% CO.sub.2 (24 h).

(14) Day 2: chemical treatment for 24 h

(15) When the cells reach about 80% confluence, they are treated with samples of the following compounds or mixtures of compounds, tested at the concentrations described hereafter: Carnitine 10 mg/ml Caffeine 10 mg/ml Arginine 30 mg/ml Carnitine 10 mg/ml+Caffeine 10 mg/ml Carnitine 10 mg/ml+Arginine 30 mg/ml Caffeine 10 mg/ml+Arginine 30 mg/ml Carnitine 10 mg/ml+Caffeine 10 mg/ml+Arginine 30 mg/ml

(16) The cells treated with culture medium only were used as control.

(17) All the cells were incubated at 37 C., 5% CO.sub.2 for 24 h.

(18) After 24 hours of treatment, the cells were washed with cold DPBS and resuspended in buffer for the ATP assay with protease inhibitor (1%). The samples were then lysed by pipetting repeatedly and centrifuged to remove insoluble materials, and finally stored on ice until use.

(19) The reaction mixture for ATP was prepared by mixing ATP Assay Buffer (88%), ATP Probe (4%), ATP Converter (4%) and Developer Mix (4%). 0.5 ml of reaction mixture of ATP were added to each 24-well plate and 50 l of each sample were added. The standard curve (0.04-0.08-0.12-0.16-0.2 mM) was set as reported in the ATP-assay kit Abcam (Abcam, Cambridge, UK).

(20) The samples were incubated at room temperature for 30 minutes, in the dark. The absorbance was read at a wavelength of 570 nm with a reference filter of 630 nm in a microplate reader Biotek ELX808 using a predetermined protocol and after having correctly defined a layout of the plate.

(21) The ATP concentration was calculated by interpolating the data of the standard curve after subtraction of the blank:
[ATP] (nmol/l)=Ts/Sv

(22) where Ts is the amount of ATP from the standard curve (nmol); SV is the volume of the sample (before dilution) added to the sample wells (l).

(23) The following results were obtained, also graphically represented in FIGS. 1 and 2:

(24) Results

(25) TABLE-US-00009 Sample ATP % vs control % Control 100.00 / Carnitine 134.16 34.164 Caffeine 126.84 26.843 Arginine 132.94 32.944 Carnitine + Caffeine 121.35 21.352 Carnitine + Arginine 117.39 17.387 Caffeine + Arginine 95.73 4.270 Carnitine + Caffeine + Arginine 240.31 140.315

(26) The results show that the only combination of active ingredients capable of providing a surprising increase % in the production of cellular ATP compared to that achieved with the individual ingredients is selectively that consisting of carnitine+caffeine+arginine, for which the ATP production increases by more than 140% compared to the untreated control.

(27) The result proves to be surprising especially compared to the case of the tested pairs carnitine+caffeine, carnitine+arginine, caffeine+arginine, for which an increase % in the ATP is found which is lower than that obtained with the individual ingredients, in one case even negative. This demonstrates that the selection of the triad according to the invention is such as to provide a result of ATP activity which is certainly unexpected and abnormal in comparison with the possible combinations of the same components in pairs.

(28) The experimental data thus show an unexpected synergy behavior in the ATP activity of the selective combination carnitine+caffeine+arginine, such as to propose an advantageous use thereof in the pharmaceutical or cosmetic field for any skin and hair treatment which requires an enhanced contribution of cellular energy, through the administration of a suitable composition with directions typically defined in the industry as energy boosters.

(29) Also in view of the background art mentioned above describing a relationship between high levels of cellular ATP and inhibition of hair loss and/or stimulation of hair growth, as well as skin health, the compositions according to the present invention are particularly indicated for such uses in humans.