CA2047751A1

CA2047751A1 - Cosmetic composition for the hair, containing a glycoprotein

Info

Publication number: CA2047751A1
Application number: CA 2047751
Authority: CA
Inventors: Alain Huc; Danielle Antoni
Original assignee: Individual
Current assignee: BASF Beauty Care Solutions France SAS
Priority date: 1989-03-03
Filing date: 1990-03-01
Publication date: 1990-09-04
Also published as: EP0461189B1; FR2643909B1; FR2643909A1; AU5270890A; WO1990009778A1; JPH04503807A; EP0461189A1

Abstract

ABSTRACT OF THE DISCLOSURE

The invention relates to a basic composition for the preparation of a cosmetic composition constituting a composition for the hair.
This composition is characterized in that it contains a glycoprotein as active ingredient, particularly selected from fibronectin or laminin, or mixtures thereof in all proportions.
A nicotinic acid salt may also be provided to reinforce the microcirculation and/or the heparin or heparan sulfate.
Due to the invention, a particularly active cosmetic composition constituting a composition for the hair is obtained, which radically improves a new growth of hair.

Description

~77~1 cos~etic com~r~.

The presen~ inven~ion relates essentially to a cosmetic composition for the hair, containing a glycoprotein.
05 More particularly, the present invention relates to a basic composition for preparing a cosmetic composition constituting a composition for the hair 9 to the cosmetic composition constituting a composition for the hair thus prepared, to a process for the preparation of the basic composition or of the cosmetic composition and to the use of a glycoprotein, alone or in combination with other active substances such as heparin, heparan sulf~te or nicotinic aeid or its salts, for preparing a cosmetic composition for hair treatment.
Many cosmetic compositions are already kn~wn, which constitute hair lotions for the treatment of the scalp. According to certain prior solutions, such cosmetic compositions contain mucopolysaccharides alone or in combination with a placental extract. For example, document FR-A-2 574 289 describes the combination of acid mucopolysaccharides extracted from the connective tissue of animals, with a bacterial cell proteolysate, from calf serum peptides, embryonic peptides, and a placental extract obtained from animals.
However, the mucopolysaccharide used and in particular those sold under the name Trichopeptides ~ cited in Page 4, lines 17 to 21, contain only traces of or no heparin or heparan sulfate. The same applies to the placental extracts of bovine origin described in said document (page 4, lines 8 to 11 and 15-16).
Other documents describing the incorporation of mucopolysaccharides to hair treatment csmpositions in~lude EP-A-035 919~ Among mucopolysaccharides, there can be cited the 4-chondroitin sulfate (page 2, lines 14 15).
FR-A-2 588 756, FR-A-2 591 889, FR-A-1 582 828, JP-A-59-186 911, JP-A-103-150 210, EP-A-182 756 and EP-A-2 094 109.
However, in none of the prior art documents, is there any heparin

2 0 ~ r3 or heparan sulfate in any significant quantity.
It is possible to explain the use of ~ucopolysaccharides recommended by the prior art documents by the fact that they improve blood circulation in the hair follicle due to a greater 05 blood fluidity and to an influence on the reconstitution of microvessels, the glycosaminogly~ans having angiogenetic properties. The mucopolysaccharides also have an influence on the development and differenciation of the cells producing hair keratine. Also, the sulfated glycosamonoglycans have an activity as a sulphur donor during synthesis of the hair keratin.
Compositions are also knawn from document EP-A-297 455, which have a hair-stimulation activity, which compositiDns comprise mixturss essentially consisting of sulfomucopolysaccharides with a predetermined field of compositions of individual glycosaminoglycans among which the following are cited in the claims, heparin between 2 and 30~ by weight, or heparan sulfate between 10 and 40% by weight.
Moreover, doeument EP-A-279 244 of TONFER describes a composition for treating and promoting hair growth comprising as active ingredients a compound selected from heparin and/or ~itamin K and/or clofibrate, the heparin content being 0.1% maximum (page 5 lines 54-56).
It is the object of the presPnt invention to solve the new technical problem consisting in providing a solution for further improving the efficiency of the cosmetic compositions constituting hair trestment compositions, notably by improving the cell davelopment and tissues regeneration hence by improving ths keratin of the ha~rO
A furthnr object of the present invanti9n is to solvu th~
new technioal problem consisting in providing a solution for prolonging the eff~ct of the ac~ive ingredients in th~ system by further improving the efficiency of the treatment, and by improvin~
adhesion between the constituents of the composition ~nd

3 2~7~3:~

the cell membranes, thereby reinfor~ing tne cell development of the cells in the hair bulb through causing an increased synthesis of keratin.
Yet a further object of the present invention is to solve 05 the new technical problem consisting in providing a sDlution for improving the availability of the acti~e ingredients and for promoting microcirculation in the hair bulb.
All said technical problems are solved for the first time in a particularly easy and inexpensive way, usable at industrial level.
Accordingly, in a first aspect, the present invention covers a basic composition for the preparation of a cosmetic composition constituting a composition for the hair, comprising at least a glycoprotein, characterized in that it further comprises collagen or atelocollagen.
According to yet another advantageous variant of embodiment of the in~ention, the glycoprotein content is comprised between about 10 4 % and 10 1 % by weight of the final basic composition.
According to another particular variant of embodiment, the glycoprotein concentration is comprised between 0.5 and 2% by weight with respect to the csllagen or atelocollagen.
According to a particular variant of embodiment of the invention, said glycoprotein is selected from fibronectin, laminin, or mixtures thereof in all proportions.
According to a particular embodiment, the fibronectin is obtained from plasma. The laminin, on the other hand, is advantageously obtained from the placenta.
According to another advantageous characteristic of the invention, the composition is characterized in that it ccmprises at least an active ingredient selected from the group consisting of heparin or heparan sulfate, preferably, between 1 and 5% by weight of the final basic composition.
According to a particularly advantageous embodiment of the invention, the composition is characterized in that it com-

4 ~ 7.
prises a microcirculation ~ctivating age~t, particularly a nicotinic acid salt such as methyl m co~inate, advantageously at a concentra-tion comprised between about 1 and 5% by weight of the final basic composition.
05 Accordlng to an~ther particularly advantageous embodiment, said compssition further comprises one ~r more mucopolysaccharides or glycosaminoglycans, the proportion of heparin or heoaran sulfate relatively to the overall content of mucopolysacchrides or glycosaminoglycans being at ieast 20% by weight.
According to another particularly advantageous embodiment of the invention, the heparin or heparan sulfate is isolated from the placenta after enzymatic digestion of the proteins by a suitable protease. A preferred protease is papain. The placenta is preferably lyophilized before being subjected to enzymic digestion with the protease. Advantageously, the ~rude extract is pur-fied so as to obtain a proportion of heparan sulfate at least equal to 40~ by we ght with respect to the total quantity of purified extract.
According to ano~her particularly advantageous embodi~ent of the inventiDn, at least one active ingredient is at least partly encapsulated in liposome type vesicles.
Acoording to an advantageous embodimen~, the liposome-type vesicles, incorporating at least partly said active ingredient, in particulr heparin or heparan sulfate, a combination of heparin or heparan sulfate in pure form or in the form of crude or purified placental extract, in the presence of glycoprotein, are stabilized by a homogeneous collagen or atelocollagen-glycosaminoglycan stabilizing solution, notably by being prepared in the presence of such stabilizing solution.
In particular, the solution is a homogeneous solution of glycosaminoglycan and atelocollagen. The glycosaminoglycan solution is prepared by placing the glycosaminoglyGan in solution in a basic aqueous solution whose pH is adjusted so that after mixing with the atelocollagen solution, the pH of the mixture constitutiny the stabilizing base remains slightly basic while remaining close to neutrality Preferably, the final pH is arwnd 8. The basic aqueous solution can advantageously be an aqueous ~77 ~

solution of sodium hydroxide~
rhe glycosaminoglycan concentration in the glycosamino-glycan solution is advantageously between 0.5 and 4%, and better still between 0.5 and 2% and still pref2rably around 1%.
05 Moreover, the atelocollagen sDlution is advantageously an aqueous solution of atelocollag0n of concentration preferably comprised between 0.5 and 2% by weight, and prefersbly still around 1%. Said solution of atelocollagen can be obtained by dissalving fibers of atelocollagen in a slightly acid aqueous solution.
According to an advantageous variant, the fibers of atelocollagen are placed in solution in acetic acid 0.1%. According ~o a variant, the atelocollagen can also be obtained by enzymic digestion of collagen. The suspension of liposome type vesicles can be introduced into the atelocollagen-glycosaminoglycan stabilizing base solution, in substantially equal volume proportions, the proporti.on of liposome type vesicles being advantageously comprised between û.5 and 2% of the final composition (expressed in proportions of lipids constituting the lipidic wall of the liposome type vesicles).
According to another variant, the liposome type vesicles are prepared in the presence of the collagen-glycosaminoglycan stabilizing base and in the presence of said glycoprotein. Said compounds enter in the constitution of the lipidic membrane of the liposomes.
Thus, according to another particularly advantageous embodiment of the invention, there is a possibility to add the glycoprotein to the atelocollagen-gly~osaminoplycanstabilizing base, the glycoprotein concentration in the stabilizing base being preferably between 0.5 and 2% by weight with respect to the collagen.
According to yet another variant of embodiment of the invention9 it is possible to use hyaluronic acid, 4-chondroitin sulfate, 6-chondroitin sulfate, dermatan sulfate, keratan sulfate or a mucoitin sulfate, as mucopolysaccharides added to heparin or heparan sulfate.

7~

According to a second aspect, the present invention further covers a cosmetic composi~;ion constituting a composition for treating the hair~ ohara~terized in that it is prepared from a basic composition such as defined hereinabove.
05 According to an advantageous variant of embodiment, the cosmetic composition is finally prepared from the basic composition by diluting the suspension of liposome type vesicles in an aqueous solution constituting a citrate buffer. Said dilution is, for example, 10 times. Preferably, an alkylene glycol, and in particular propyleneglycol, is added to said citrate buffer.
The citrate buffer, as well as the alkyleneglycol such as the propyleneglycol, improve the homogeneity of the composition for the hair. The invention makes it possible to improve beyond all expectations, the cellular development and regeneration of tissues, hence ~he keratin of the hair. The formulation in liposome type vesicles facilitates the penetration of the heparan sulfate or of the heparin when in contact with the membrane of the cells synthesizing the keratin of the hair~
Further, the synergistic effects produced by combining the atelocollagen-glycosaminoglycan alone or else with glycoprot~in, improve cell proliferation as well as the adhesion of the constituents of the composition according to the invention to the cell membrane. It has been observed that glycoproteoin has specific interaction sites with the cell membrane and with the collagen fiber. The composition accordiny to the invention thus makes it possible to promote in unexpected manner the development of the cells of the hair bulb, and to increase the synthesis of the keratin.
In addition, another synergistic effect resides in the fact that the glycoprotein, and in particular fibronectin or laminin, is stabili~ed by ~ollagen (atelocollagen) so that said glycoprotein is maintained in the native state in the composition as well as when this is applied on the hair.
Also, the use of a nicotinic acid salt, particularly in a form at least partly incorpora~ed in the liposome type vesicles, 2~ 7 ~ ~

enables the improvement of the microcirculation around the hair bulb. It is noteworthy that another particularly unexpected effect of the use of the combination of heparan sulfate or heparin, either in pure form or in the form of crude or purified placental 05 extract, with a glycoprotein, resides in the cell proliferation resulting from ths trapping of the growth factors in the hair bulb and of the collagen or atelocollagen-mucopolysaccharide-glycoprotein combination.
Other aims, characteristics and advantages of the invention will become clearer in the light of the following explanatory descrlp~ion, made with reference to the examples given hereinafter solely by way of illustration and non-restrictively.
Example 15 1) Preparation of the placental mucopolysaccharide extractcontaining heparan sulfate.
A) Reagents a) Enzyme The enzyme used is the "Merck" papain at 30 000 U.O./mg.
b) KCL buffer Its composition is as follows:
Potassium chloride 11.8 g/l Cystein 78.8 mg/l EDTA 180 mg¦l The pH is adjusted to 6.5.
B) Method The ovine placenta collected just a~ter birth is lyophilized.
30û 9 of enzyme are dissolved in 225 l of KCL buffer.
30 kg of lyophilized pl3c~nta are then introduced into this bath.
The temperature of the bath is brought to 60C and is kept to this value throughout the whole operation of enzymatic digestion After 24 hours, a quantity of en~yme equal to that used is added to the reaction bath.
A~ter another 24 hours, the bath is decanted by natural 7 ~ ~.

gravity and the supernatant is centrifuged with a Robatel centrifugal machine rotating at 3000 revs/min. and giving an accPleration sf 2000 9.
The liquid temperature reached is lowered to 15O. Then 05 a quantity of trichloroacetic acid such as to obtain a concentration of 3.5% is added to the bath kept under stirring. The resulting mixture is le~t to stand for about ten hours. The precipitate obtained by natural gravity is eliminated and the solid particles remaining in the supernatant are separated by the 10 centrifugal machine in the same conditions as those described hereinabove.
The resulting liquid is then neutralized with sodium carbonate. The bath is left to stand for 3 hours.
After decantation and centrifugation, the supernatant is 15 ultrafiltrated by means of a Millipore ultrafiltration apparatus comprising membranes of which the cut-off threshold is lO,000 Daltons. The ultrafiltrate is lyophilized. About 500 9 of placental mucopolysaccharide extract are obtained.
2) Pur~fication of the heparan sulfate 1 kg of Dowex ~J resin (1 x 2, Cl- form, 100-20û mesh) and 1.5 kg of placental extract obtained as ahove, are added to 15 liters of ion-exchanged water. After stirring for one hour, the resin is decanted by natural gravity and the supernatant is rejected. The resin is then placed in a bath containing 0.75 M
25 NaCl.
1st bath After stirring for 1 hour, the resin is decanted in a natural way and the supernatant is sent into the ultrafiltering equipment.
The obtained ultrafiltrate is lyophilized. In this way, about 150 9 of dry hyaluronic acid are prepared.
2nd bath __ The resin is then placed in a bath containing 1.6 M NaCl.
Proceeding as beore, 450 9 of dry heparan sulfate are obtained.

2 ~ 4 7 7 ~ 1 3rd bath The r~sin is finally placed in contact with a bath containing 3 M of NaCl. ProceRding as before, 30 9 of d0rmatane sulfate contaminated by a small quantity of heparan are obtained 05 in dry form.
3) Preparation of decrDsslinked collagen The skin of freshly slaught0red calf is subjected tu chemical depilation in a b3th containing 3~ sodium sulfide and 4%
lime, the proportion being 100 9 of skin for 200 cm3 of solution.
The derm is then isolated from the rest of the skin by a slitting operation using a rotary band saw~
The obtained tissue is ground and extruded through a screen having 4 mm-holes. The ground material is then placed, for thrPe weeks in contact with saturated lime milk in the proportion of 1 kg for 4 l of solution. The skin, having been treated in this way, is separated from the supernatant by continuDus centrifuging under an acceleration of 2000 9 using a centrifugal machine rotating at 4000 revs/per minute. The concentrate then undergoas two washes in running water in a stainless steel tank under slow stirring, in the proportion of 1 kg for every liter of bath. Then, the ground material undergoes two treatments with the phosphate buffer pH 7.8 (21~7 g/l of Na2HP04 and 0.78 9/l of KHzP04) in the same conditions as for the wash in water. The residue is then washed in two baths of demineralized and sterile water. The resulting ground material is placed in a solution of acetic acid (0.5 9/1, pH 3, 4) in the proportion of 1 kg for 20 l of bath.
After stirring for 5 mins., the supernatant is separated ~rom the residue by continusus centrifuging according to the afor0said technique. The collagen is then precipitated by the supernatant by addition of dry sodium chloride in thP- proportion of about 10%
with respect to the bath. After gravity-decanting, the obtained fibers are dialyzed against demineralized and sterile water using dialysis membranes, preferably formed by guts whose cut-off threshold is oetween 6000 and 8000 Daltons. After checkins with silver nitrate ~hat the dialyzed fibers no longer contsin sodium 2~ J7^J3~

chloride~ these are placed in solution in a bath containing 6 g/l of acetic acid so as tD obtain a final protein concentration of 1%. The mixture is stirred slowly for 24 hours. About 20 l of solution being obtained ~rom 1 kg of ground material.
0~ 4) Preparation of 4-chondroitin sulfate Nasal septa from calves, from which the muscle and adipose tissues have been removed, are chopped and ground by extrusion through a screen having 4 mm holes; the ground material i5 placed for 24 hours at a temperature of 6C in a potassium chloride buffer (11.8 9/1 of KCl, 78.8 mg/l of cystein, ETDA lB0 mg/l) containing 1% of "Merck" papain. The proportion of ground material being 130 g for 1 liter of buffer.
The supernatant is separated from the residue by continuous centrifuging using a centrifugal machine turning at 4000 revs/min. Then, 40 9/1 of trichloroacetic acid are added to the supernatantO The precipitate is removed by con~inuous centrifuging according to the aforesaid technique. The supernatant is neutralized with pellets of sodium carbonate. The mixture is then dialyzed against demineralized and sterile water, using guts whose cut-off threshold is comprised between 6000 and 8000 Daltons. The dialyzed solution is lyophilized. The 4-chondroitin sulfate is obtained in dry form. 90 9 of mucopolysaccharides are obtained with 1 kg of nasal septa.

5) Preparation of a composition for the hair A) Basic liposome solution In 10 kg of complex of collagen glycosaminoglycan containing 20 9 of collagen, 5 9 of 4-chondroitin sulfate and preserving agents, such as 10 9 of Nipagi ~ and 50 9 of Phenonip~
from NIPA Lab.Ltd/UK, are added for example 10 g of heparan sulfate and 10 9 of methyl nicotinate. The mixture is stirred mechanically until a homogeneous solution is obtained, the pH is adjusted to 7.2.
100 9 of a homogeneous mixture of lecithin and cholesterol are added to the bath, kept under stirring at room temperature. After complete dissolution, the mixture is stirred ~1 2~77~

with ultrasonic stirring means of Ultra Turax UTL T60 type at 8000 revs/min. for lû mins., then 0.2% methyl nicotinate and 0.002%
native fibronectin extracted from bovine plasma are added. The mixture is homogenized under slight stirring.
05 B) Final cOmpGsition for the hair 10 kg of the above liposome solution are diluted in 50 liters of sterile ion-exchan~ed water. Then, 5 kg of propyleneglycol, lOû g of Nip3gin~ and 500 9 of Phenoni ~ as well as 200 g of sodium triacetate are added to the solution. The volume of the bath is then completcd to make up 100 liters and the pH is adjusted to 7.8 with citric acid.
The resulting composition is slightly cloudy, opalescent and has a yellowish white color.
Example 2 The methodology is exactly the same as before, except that the pure heparan is replaced by the placental mueopolysaccharide extract. But in this case, the quantity of liposome solution will be doubled. This means that its concentration in the final composition will be 20% instead of 10%
as before.
_ample 3 The composition for the hair is the same as that of Example 1, except that the hepar~n sulfate is replaced by heparin.
~xample 4 NEW GRûWTH OF HAIR TESTS
The new growth of hair was ~tudied on rats treated with the composition prepared according to Example 19 in comparison with two animals treated with placebo, namely with the mixture of citrate buffer, propyleneglycol and preserving agent.
To this effect, 20 cm2 of the skin on the backs of the rats, previously shaved, were treated daily with 0.05 9 of either the composition according to the invention, or of placebo. After 21 days, it was found th3t the new growth of hair was in average twice as quick in the rats treated with the lotion. This test was conducted on two batches of 10 rats.

12 2 Q 4 7 7 ~ 1 The product can be packed in ampules of 5 ml. This quantity corresponds to a daily treatment.
The invention further covers the processes for preparing the basic composition or the cosmetic composition, characterized in 05 that they include incorpora~ing at least one acti~e substance such as def.ined hsreinabove in a suitable vehicle, support or excipient, and in particular a cosmetically acceptable vehicle, support or excipient.

3~

Claims

The embodiments of the invention in which an exclusive property or privilege is claimed, are defined as follows:

1. Basic composition for the preparation of a cosmetic composition constituting a composition for the hair, comprising at least a glycoprotein, characterized in that it further comprises collagen or atelocollagen.

2. Composition according to claim 1, characterized in that the glycoprotein content is comprised between about 10-4% and 10-1% by weight of the final basic composition.

3. Composition according to claim 1 or 2, characterized in that the glycoprotein concentration is comprised between 0.5 and 2% by weight of the collagen or atelocollagen.

4. Composition according to one of claims 1 to 3, characterized in that said glycoprotein is selected from fibronectin, laminin, or mixtures thereof in all proportions.

5. Composition according to claim 4, characterized in that the fibronectin is obtained from plasma.

6. Composition according to claim 4, characterized in that the laminin is obtained from placenta.

7. Composition according to one of claims 1 to 6, characterized in that at least one active ingredient is at least partly encapsulated in liposome type vesicles.

8. Composition according to claim 7, characterized in that the liposome type vesicles, incorporating at least partly one active ingredient, are stabilized by a homogeneous stabilizing solution of collagen or atelocollagen-glycosaminoglycan, containing at least partly said glycoprotein, notably by being prepared in the presence of said stabilizing solution.

9. Composition according to claims 1 to 8, characterized in that it comprises at least one active ingredient selected from the group composed of heparin or heparan sulfate, preferably, the heparin or heparan sulfate content is comprised between about 1 and 5% by weight of the final basic composition.

10. Composition according to one of claims 1 to 9, characterized in that it further comprises a microcirculation activating agent, notably a nicotinic acid salt, such as methyl nicotinate, advantageously in a concentration comprised between about 1 to 5% by weight of the final basic composition.

11. Composition according to claim 9 or 10, characterized in that it further comprises one or more mucopolysaccharides or glycosaminoglycans, the heparin or heparan sulfate content relatively to the total content of mucopolysaccharides or glycosaminoglycans being at least 20% by weight.

12. Composition according to one of claims 9 to 11, characterized in that the heparin or heparan sulfate is isolated from the placenta after enzymatic digestion of the proteins by a suitable protease, preferably papain, in pure form or in the form of pure or purified placental extract.

13. Composition according to claim 12, characterized in that the placenta is lyophilized before undergoing the enzymatic digestion with the protease; preferably the crude extract is purified so as to obtain a heparan sulfate proportion at least equal to 40% by weight with respect to the total quantity of purified extract.

14. Composition according to one of claims 8 to 13, characterized in that the suspension of liposome type vesicles is associated to the stabilizing base solution of atelocollagen-glycosaminoglycan in substantially equal volumes, the proportion of liposome type vesicles being preferably comprised between 0.5 and 2% by weight of the final basic composition.

15. Composition according to one of claims 11 to 14, characterized in that hyaluronic acid, 4-chondroitin sulfate, 6-chondroitin sulfate, dermatan sulfate, keratan sulfate or a mucoitin sulfate is used as mucopolysaccharides.

16. Cosmetic composition constituting a composition for the treatment of the hair, characterized in that it is prepared from a basic composition such aa defined in any one of the preceding claims.

17. Cosmetic composition according to claim 16, characterized in that it is finally prepared from the basic composition by diluting the suspension of liposome type vesicles in an aqeuous solution forming a citrate buffer, in which is preferably added an alkyleneglycol, in particular the propyleneglycol, particularly diluted about 10 times.

18. Process for the preparation of a basic composition or of a cosmetic composition, characterized in that it comprises incorporating at least one active ingredient such as defined in any one of claims 1 to 17, in a suitable vehicle, base or excipient, particularly in a suitable cosmetically acceptable vehicle, base or excipient.

19. Use of a glycoprotein, notably fibronectin or laminin, in combination with collagen or atelocollagen, for the preparation of a basic cosmetic composition or of a cosmetic composition for the treatment of the hair.