FR2682039A1 - Medicational compositions intended to promote regeneration of the extracellular matrix - Google Patents

Abstract

These medicational compositions contain intracellular (subcellular) vesicular vectors (carriers) including metabolic ligands capable of being expressed inside and outside their structure. These vectors are intracellular vesicular vectors with specific affinity for the fibroblast whose membrane composition, modified by the introduction of metabolic ligands chosen from the initiators and regulators of cell synthesis which are endowed with a lipophilic radical or functional group by acylation, brings about a regulation in the activity of synthesis of the target cells. They are chosen from artificial spheres, capsules or vesiculas having a diameter which can range from 3 to 1000 nanometers, used in order to incorporate an active substance either into an aqueous medium which fills the internal volume of the vector or in the wall itself of the said vector, and then in order to transport this active substance to a site at which it will be released and used towards various ends. These medicational compositions find applications in increasing the synthesis of collagenases with a view to resorbing dense fibrous tissues of collagen I or cheloids, as well as in directing the synthesis of collagen towards a specific structure such as collagens I and III.

Description

MEDICAMENT COMPOSITIONS FOR PROMOTING
RENEWAL OF THE EXTRACELLULAR MATRIX
The present invention relates to medicinal compositions for promoting the renewal of the extracellular matrix.

 The abundance of the extracellular matrix that separates cell constituents is known to be one of the characteristics of connective tissue. This matrix is at the origin of various functions fulfilled by this fabric. It serves as a biomechanical support element and its varied composition influences cellular behavior.

 Far from being a stable structure, this matrix is constantly reworking because of the cellular activity that synthesizes and modifies it. Changes occurring in the extracellular matrix may be of a quantitative or qualitative nature. An imbalance between synthesis and / or degradation may contribute to the establishment of a "modified" or pathological extracellular matrix.

 It is known that a very large variety of molecules that make up the extracellular matrix (collagens, proteoglycans and glycoproteins) are synthesized by fibroblasts.

 Collagen is one of the major components of the extracellular matrix. There are currently 13 types of collagen from a family of twenty genes. These molecules are formed of a combination of three peptide chains in triple helix alpha. In the helical part, an amino acid sequence is repeated, where glycine often precedes proline and hydroxyproline.

 The specific protein structure allows collagens to perform different functions within the connective tissue.

 Of the different types of collagen, type I and type III collagens are the major fibroblast synthesis product. Emonard and Grimaud (Cellular and Molecular Biology, 36 (2) 131-153 - 1990) have shown that fibroblasts can control the quality and quantity of collagens not only by decreasing synthesis, but also by synthesizing enzymes. corresponding degradation. . Thus, the fibroblastic cell alone can control the entire connective matrix deposit. This fibroblastic cell is furthermore permanently influenced by soluble factors (cytokines) and by insoluble factors (matrix components).

The result of these interactions conditions the fibroblastic function towards
an activity of synthesis or degradation with respect to the extracellular matrix.

 Fibroblasts are therefore a fundamental part of the connective tissue physiology and the control of its functions is of paramount importance in the medical field.

 The control of collagen synthesis by fibroblasts is of major importance for connective tissue physiology. Indeed, the fibroblast is the mesenchymal cell responsible for the synthesis, organization and remodeling of the conjunctive matrix.

 Among the different molecules synthesized by fibroblasts, we have seen that type I and type III collagens are quantitatively the most important.

As a result, the use of fibroblast culture makes it possible to accurately analyze the action of the various molecules on the activity of collagen synthesis or degradation.

 A large number of drugs have already been described as being capable of modifying fibroblastic function (Bissel and Coll, Hepatology, No. 3,1990 p.

488-498 - Emonard and Grimaud, in Cellular and Molecular Biology, 36 (2) 131-153 1990 - Kovacs in ImmunologyToday Vol. 12, N, 1991).

 Some of these are capable of stimulating the synthesis of collagen by fibroblasts: ascorbic acid (Chojkier et al., Jal of Biological Chemistry - Oct. 5, 1989, pp. 16957-16962), cytokines such as interleukin 1, platelet derived growth factor (PDGF) (Kovacs - Immunology Today 1 - 1991), collagen-derived peptides (Pacini et al, Res.Comm.In Chemical Pathology and Pharmacology - April 1990 pp. 89-101) .

Other molecules have been studied to inhibit collagen synthesis and sometimes induce the synthesis of degradation enzymes (collagenases). The best known are corticosteroid hormones, cytokines such as tumor necrosis factor (TNF) and interferons; see: Bissel and Coll, Emonard and Grimaud,
Granstein et al. (The Jal of Investigative Dermatology Suppl.1990) and Kovacs).

 It is known that the synthesis of collagen requires a large intake of amino acids.

 The patent application filed on the same day "Vesicular infracellular vectors and their application in cosmetic compositions" describes and claims certain infracellular vesicular vectors having metabolic ligands able to express themselves inside and outside their structures.

 The inventors have been able to determine that the abilities presented by all these vesicular vectors in the control of collagen synthesis by fibroblasts enabled them to find particularly useful applications in the production of medicinal compositions intended to promote the renewal of the matrix. extracellular.

 Indeed the use of these vesicular infracellular vectors makes it possible to add to the culture medium nutrients having no cytotoxic effect for the cells.

 The invention thus relates to medicinal compositions intended to promote the renewal of the extracellular matrix, characterized in that they contain subcellular vesicular vectors comprising metabolic ligands able to express themselves inside and outside their structure.

 According to a preferred embodiment of the invention, the vectors are infracellular vectors of specific fibroblast affinity whose membrane composition, modified by introduction of metabolic ligands, leads to a modulation of the synthesis activity of the targeted cell.

 These metabolic ligands are preferably chosen from initiators and modulators of cell synthesis with a function or lipophilic radical by acylation, such as the fatty esters of ascorbic acid such as its palmitate, collagen hydrolyzate palmitate, esters. fatty acids of hydroxyproline and more especially its palmitate, and the peptides derived from elastin.

 Ascorbic acid is in particular immobilized within the vector in the form of a fatty acid ester, which prevents any leakage of the compound.

 The hydrophilicity of the vector membrane is obtained by the attachment of fatty amide (lipoamino acid) to the outside of the vector.

 We see all the benefits of transport by the same vector of a stimulant of the synthesis of modified collagen for better encapsulation and precursors of collagen synthesis.

 In the present description, the term vesicular vector (or, to simplify vector ") will apply to all vesicles, capsules or artificial spheres of a diameter that may range from 30 to 1000 nanometers, and which are used to incorporate a substance active either in an aqueous medium filling the interior volume of the vector or in the same wall of said vector and then to transport this active substance to a site where it will be released and used for different purposes.

 According to the invention, the use of the vesicular vectors mentioned above as vectors of bioactive molecules able to stimulate the synthesis of collagens by fibroblasts or to inhibit this synthesis and to induce the synthesis of degradation enzymes considerably strengthens or modifies the action of these molecules; they also act as modifiers of their cytotoxicity which is of considerable interest in their application as drug compositions intended to promote the renewal of the extracellular matrix and to accelerate healing.

 In the experiments described hereinafter, ascorbyl palmitate, peptides derived from collagen, and hydroxyproline palmitate have been used more specifically.

The results of the tests presented below were obtained according to the following methodology:
Culture conditions: fibroblasts are obtained from human skin explants. The cells are maintained in culture in DMEM medium (Gibco) supplemented with 10% fetal calf serum (Gibco) and antibiotics (penicillin 100 U / ml, streptomycin 100 mg / ml). The subcultures are made by trypsin (Trypsin-EDTA, Gibco). The cells are used from the 2nd to the 10th passage.

 Treatment conditions: The control fibroblast cultures refer to the cultures with vectors without active principle, in a quantity similar to the other treatments. The cultures treated with the molecule in solution refer to the conditions of the control culture and the molecule in soluble form. Cultures treated with the vector form of the molecule imply that the active molecule is previously included in the vectors.

 Cell proliferation test (MTT test): after inoculation, the cells are incubated for three days with medium containing the test molecules.

Then the yellow tetrazolium salt (5 mg / ml) is added.

It is reduced by mitochondrial reductases of living cells to a purple formazan product which is then solubilized with 0.04 N HCl in isopropanol. The color reaction is read at a wavelength of 570 nm by a
Multiskan Titertek (Flow Laboratories) and expressed in optical density.

 Evaluation of Soluble Collagens by Radioimmunoassay: 100 μl of culture supernatant are withdrawn and 100 μl of type I anti-collagen or type III anti-collagen antibody and 200 ml of PBS are added. After 24 hours at 4 ° C., 100 μl of type I or type III labeled collagen 125 are added; the antigen bound to the antibody is then separated from the free antigen by precipitation with 100 ILI of rabbit anti-IgG goat antiserum in the presence of polyethylene glycol (PEG) 6000.

 After centrifugation, the radioactivity of the precipitate is measured using a LKB 1260 multigamma counter.

Measurement of collagenase activity in the culture medium
Interstitial collagenase, which degrades native HII interstitial collagens, is measured.

a) Preparation of the substrate: the carbon-14 labeled type I collagen dissolved in 0.1M CH3COOH is neutralized to pH 7.4 with 1 M Tris and diluted with 0.15 M NaCl / CaCl 2 buffer , 4 mM / 50 mM Tris-HCl, pH 7.4, containing
0.5 mM NEM and 0.1 mM PMSF at 10,000 cpm / 100Z1 substrate (= 10000 cpm / assay).

 b) Assay of collagenase 100 μl of medium are activated with 10 μl of trypsin (1 mg / ml) at 37 ° C. for 10 min, followed by the addition of 10 μl of SBTI (5 mg / ml): 10 min at ambient temperature. 100 l of medium are also tested without trypsin activation (measurement of the active form of the enzyme). The substrate (10000 cpm / test) is added, optionally in the presence of 25 mM EDTA, the final volume being 250 μl.

 The reaction is carried out for 18 h at 25 ° C. and is then stopped by addition of ILI of 250 mM EDTA (10 min at room temperature). The mixture is then heated for 10 minutes at 39 ° C. The non-degraded collagen is precipitated with ethanol at the final concentration of 18% (30 min at room temperature). Finally, the mixture is centrifuged at 6000 g for 30 minutes at 4 C.

 200 μl aliquots of the supernatants are mixed with 5 ml of aqueous scintillant liquid and counted for their radioactivity. A collagenase unit is defined as the amount of enzyme required to degrade 1 g of native collagen per hour at 25 C.

 The study of the influence of the vectors according to the invention will now be described in detail for each of the molecules.

PALMITATE OF ASCOR BYLE
The incorporation of ascorbyl palmitate into the vectors potentiates its action on collagen synthesis by fibroblasts. Indeed, half the dose of ascorbyl palmitate (12.5 mg / ml) incorporated into the vectors according to the invention is capable of inducing the same level of type I collagen synthesis as twice its dose (25%). mg / ml) in soluble form.The effect of ascorbyl palmitate incorporated into the vectors according to the invention is even stronger on the synthesis of type III collagen. The results are summarized in Table 1 below.
TABLE 1
Stimulation of collagen synthesis by ascorbyl palmitate after 48
hours of culture
Collagen I Collagen III
ng / ml ng / ml
Fibroblasts + vectors 67 # 13 <20
Fibroblasts + ascorbyl palmitate in solution 12.5 mg / ml 668 a, b # 71 468 a, b # 41
Fibroblasts + vectors loaded with ascorbyl palmitate 12.5 mg / ml 950 a, b # 58 882 a, b 64
Fibroblasts + ascorbyl palmitate in solution 25 mg / ml 952 a, b + 81 781 a, b # 55
Fibroblasts + vectors loaded with ascorbyl palmitate 25 mgA 1275 a, b 114 1035a, be
a Significant (p <0.05) compared to fibroblasts + vectors (control)
b Significant (p <0.05) relative to the soluble molecule.

 These results show the whole point of the use of the vesicular infracellular vectors according to the invention for the preparation of medicinal compositions intended to promote the deposition of a new extracellular matrix by a stimulating effect of collagen synthesis. The incorporation of ascorbyl palmitate into the vectors according to the invention makes it possible to obtain a similar effect with a smaller amount of active ingredient.

COLLAGEN HYDROLYZATE PALMITATE
Collagen peptides in the form of palmitate have been used to stimulate the synthesis of type I and III collagens as well as the synthesis of interstitial collagenase (MMP-1).

It has been observed that the hydrolyzate of collagen incorporated in the vectors according to the invention is less toxic than in soluble form. Indeed, cytotoxicity is detected from a dose of 20 mg / ml for the soluble molecule (Table 2)
TABLE 2
Analysis of the proliferation of fibroblasts treated with hydrolyzate palmitate
collagen
Form 'igImI
1 5 10 20 40 80 24 hours
Solution 0.486 # 0.026 0.481 * 0.009 0.450 # 0.009 0.290ai 0.0110.093a i 0.009 0.037ai 0.004
Vector 0.468 i 0.019 0.476 i 0.014 0.478 i 0.012 0.487Ci 0.013 0.496c # 0.015 0.493C i 0.011 48 hours
Solution 0.488 i 0.012 0.474 # 0.018 0.470 i 0.008 0.138ai0.017 0.058a # 0.010 0.033a # 0.008
Vector 0.493 i 0.017 0.489 i 0.007 0.494 i 0.008 0.4980i0.017 0.4920i0.013 0.0482c # 0.006 72 hours
Solution 0.491 i 0.010 0.492 i 0.021 0.466 i 0.019 0.144a # 0.007 0.043a # 0.003 0.023ai 0.005
Vector 0.497 i 0.017 0.489 i 0.016 0.491 i 0.011 0.4780i0.012 0.502c # 0.021 0.479 i 0.009
The data are expressed in measured optical density after treatment on cultures containing 104 cells per well.

a significant (p <0.05) compared to the lowest doses c significant (p <0.05) compared to the molecule in soluble form
With regard to collagen synthesis, it was noted that starting from a dose of 5 mg / ml, the collagen hydrolyzate significantly stimulates the synthesis of type I collagen. This effect is observed using the molecule incorporated into the vectors (Tables 3 and 4).

TABLE 3 Analysis of the synthesis of collagen type I by fibroblasts treated with the
collagen hydrolyzate palmitate
Form g / ml
1 5 10 20 40 80 24 hours
Witness 274 # 34,353 # 22,228 # 8 187a # 10 160a # 15 135a # 10
Solution 274 # 28 211a # 12 203a # 13 63a, b # 2 49a, b # 13 30a, b # 3
Vector 239 # 23,297 # 28 213 # 20 229c # 21 225b, c # 27 221b, c # 5 48 hours
Witness 406 # 37 360 # 11 299a # 27 258a # 4 207a # 5 183a # 5
Solution 454 # 65,358 # 13 272a # 11 80a, b # 5 31a, b # 8 41a, b # 4
Vector 536a # 32 502b, c # 26 470b, c # 48 357a, b, c # 15 330a, b, c # 16 304a, b, c # 24 72 hours
Witness 454 # 37,383 # 35,379a # 40 246a # 3 232a # 8 219a # 5
Solution 456 # 29 353a # 9 302a # 5 97a, b # 5 19a, b # 5 14a, b # 3
Vector 601 # 128 577b, c # 41 457b, c # 13 352a, b, c # 20 260a, b, c # 23 325a, b, c # 10
The data are expressed in ng / ml a significant (p> 0.05) compared to the lower doses b significant (p> 0.05) relative to the control c significant (p> 0.05) with respect to the molecule in soluble form
TABLE 4
Analysis of the synthesis of type III collagen by fibroblasts treated with the
colloidal hydrolyzate palmitate
Form g / ml
1 5 10 20 40 80 24 hours Witness 258 # 57 253 # 43 183 # 18 183 # 30 127a # 22 49a # 3
Solution 318 # 67 292 # 25 277b # 17 25a, b # 1 23a, b # 2 19a, b # 1
Vector 309 # 45 311 # 12 261c # 9 255c # 34 211b, c # 29 207a, b, c # 20 48 hours Witness 498 # 74 260 # 15 206a # 30 210a # 10 183a # 21 106a # 6
Solution 744 * 123 492a, b # 312a, b 23a, b * 1 20a, b # 3 25a, b * 2
Vector 780b # 76 677b, c # 12 665b, c # 120 518a, b, c # 29 382a, b, c # 48 309a, b, c # 26 72 hours
Witness 507 # 30 268a # 8 182a # 37 163a # 12 173a # 29 128a # 1 Solution 569 # 61 446b # 39 352a, b # 44 31a, b # 3 18a, b # 1 15a, b # 2
Vector 680b # 39 793b, c # 77 380a, b, c # 47 415a, b, c # 33 352a, b, c # 20 342a, b, c # 22
The data are expressed in ng / ml a significant (p> 0.05) compared to the lower doses b significant (p> 0.05) relative to the control c significant (p> 0.05) with respect to the molecule in soluble form
The synthesis of type III collagen is stimulated with a dose of 10 mg / ml and only when the collagen hydrolyzate is incorporated into the vectors according to the invention.

 This same treatment significantly stimulates the synthesis of interstitial collagenase (MMP-1) whether in soluble form or in vector form. However, it should be noted that stimulation is significantly stronger when the molecules are incorporated into the vectors (Table 5).

TABLE 5
Dosaae of the activity of interstitial collagenase (MMP-1) produced by
fibroblasts treated with collagen hydrolyzate Palmitate
Lig / ml form
1 5 10 Solution 1551a # 110 1158a # 152 1144a # 41
Vector 2372a, b * 328 2463a, b * 1 67 2050a, b * 87
Witness: 438 * 82
These data are expressed in mU / ml of culture supernatants with 4 × 10 5 cells and measured after 48 hours of treatment.

a significant (p <0.05) relative to the control b significant (p <0.05) compared to the molecule in soluble form.

 The above results reveal all the advantage of the vector form for obtaining drug compositions that promote renewal of the extracellular matrix by concomitant activities of synthesis of new molecules (collagen) and degradation of the preexisting matrix (collagenase). ). The molecules incorporated in the vectors are less toxic. These drugs may find application in the case of hypertrophic or keloids scars that are currently treated with cytokines (Granstein et al., The Jal of Investigative Dermatology Supl Dec 1990, 75 S-80 S) (Kovacs). - immunology Today 1 - 1991).

HYDROXYPROLINE PALMITATE
Treatment of fibroblast cultures with hydroxyproline palmitate shows that the soluble form is cytotoxic from 20 mg / ml.

 However its incorporation into the vectors eliminates this cytotoxic effect even at a dose of 80 mg / ml (Table 6).

TABLE 6
Analvse of proliferation of fibroblasts treated with Palmitate
hydroxyproline
Form g / ml
1 5 10 20 40 80 24 hours
Solution 0.493 # 0.029 0.492 # 0.016 0.502 # 0.019 0.457 # 0.022 0.147a # 0.019 0.060a # 0.022
Vector 0.483 # 0.026 0.479 # 0.029 0.485 # 0.033 0.502 # 0.037 0.487c # 0.019 0.485c # 0.023 48 hours
Solution 0.494 # 0.012 0.513 # 0.025 0.511 # 0.020 0.471 # 0.028 0.118a # 0.018 0.060a # 0.016
Vector 0.493 # 0.021 0.495 # 0.025 0.487 # 0.035 0.519 # 0.024 0.545c # 0.021 0.556a, c # 0.022 72 hours
Solution 0.490 # 0.019 0.500 # 0.028 0.496 # 0.017 0.414a # 0.010 0.063a # 0.021 0.033a # 0.012
Vector 0.505 # 0.018 0.515 # 0.017 0.406 # 0.020 0.489c # 0.015 0.495c # 0.018 0.520c # 0.022
The data are expressed in measured optical density after treatment on cultures containing 104 cells per well a significant (p <0.05) compared to lower doses c significant (p <0.05) compared to the molecule in soluble form
Collagen synthesis type I and III is significantly stimulated by the vector form of the molecule at a dose of 10 mg / ml (Tables 7 and 8)
TABLE 7
Analysis of the synthesis of type I collagen by fibroblasts treated with the
Hydroxyproline Dalmitate
Form g / ml
1 5 10 20 40 80 24 hours
Witness 246 # 46 275 # 22,292 # 3,259 # 8,221 # 11 145a # 12
Solution 268 # 18 232 # 10 166a, b # 15 128a, b # 4 61a, b # 8 41a, b # 2
Vector 266 # 10 288c # 14 253c # 11 251c # 16 184a, c # 31 164a, c # 7 48 hours
Witness 487 # 67,518 # 0 449 # 57,392 # 22,345 # 18,298a # 15
Solution 519 # 37 360a, b # 42 260a, b # 7 227a, b # 1 16a, b # 6 49a, b # 10
Vector 528 # 30 605c # 43 432c # 69 421a, c # 9 422a, b, c # 26 401a, b, c # 18 72 hours Witness 431 # 8,446 # 15 446 # 52,499 # 39,368a # 18,297a # 5
Solution 513 # 77 ## EQU1 ##
Vector 483 # 46 516 # 78 524c # 17 526c # 90 387a, c # 10 354a, b, c # 20
The data are expressed in ng / ml a significant (p> 0.05) compared to the lower doses b significant (p> 0.05) relative to the control c significant (p> 0.05) with respect to the molecule in soluble form
TABLE 8 Analysis of the synthesis of collagen type III by fibroblasts treated with the
hydroxyproline palmitate
Form zg / ml
1 5 10 20 40 80 24 hours Witness 239 # 33 221 # 31 224 # 17 211 # 14 137a # 9 81a # 10
Solution 295 # 24 281 # 16 15a, b # 13 105a, b # 6 25a, b # 2 24a, b # 4
Vector 272 # 21 292b # 2 290b, c # 16 313b, c # 22 215a, b, c # 14 119a, b, c # 10 48 hours
Witness 624 # 83 528 # 47 376a # 36 385a # 14 369a # 23 310a # 14
Solution 588 # 69519 # 20 330a # 35 253a, b # 18 36a, b # 5 18a, b # 3
Vector 699 # 117 702b, c # 14 509b, c # 65 525b, c # 14 532b, c # 91 494b, c # 20 72 hours Witness 452 # 41 450 # 46 405 # 49 385 # 18 313a # 29 280a # 18
Solution 628 # 161 548 # 24 301a # 23 240a, b # 11 29a, b # 1 24a, b # 1
Vector 519 # 108 558b # 7 616b, c # 91 637b, c # 38 444b, c # 27 418419b, c # 28
The data are expressed in ng / ml a significant (p> 0.05) compared to the lower doses b significant (p> 0.05) relative to the control c significant (p> 0.05) with respect to the molecule in soluble form
The collagenolytic activity of fibroblasts is modulated by the use of hydroxyproline palmitate and its effects are related to the form of vector or soluble use. The soluble form significantly stimulates the synthesis of collagenase even with a dose of 1 mg / ml (Table 9).

 On the other hand, the use of the vector form of hydroxyproline palmitate gradually reduces, in a dependent manner, the synthesis of collagenase (Table 9).

TABLE 9
Assay of interstitial collagenase (MMP-1) activity produced by
fibroblasts treated with hydroxyproline palmitate
Form Fg / ml
1 5 10
Solution 1273ai 154 1934ai49 2189ai 118
Vector 1648a, b # 166 1090a, b 66 604a * 101
Witness: 438 + 82
These data are expressed in mU / ml of culture supernatants with 4 × 10 5 cells and measured after 48 hours of treatment.

a significant (p <0.05) relative to the control b significant (p <0.05) compared to the molecule in soluble form.

 The vector form makes it possible to obtain medicinal compositions capable of promoting the renewal of the extracellular matrix by an effect stimulating the synthesis of collagen by fibroblasts. The incorporation of ascorbyl palmitate into the vectors allows better efficacy at low doses.

 The medicinal compositions according to the invention find, inter alia, applications for increasing the synthesis of collagenases with a view to resorbing dense fibrous tissues in collagen I or keloids, as well as the orientation of collagen synthesis. to a particular structure such as collagens I and III.

Claims (9)

 1 - Drug compositions intended to promote the renewal of the extracellular matrix, characterized in that they contain infracellular vesicular vectors comprising metabolic ligands able to express themselves inside and outside their structure.  2 - drug compositions according to claim 1 characterized in that the vectors are infracellular vectors of specific fibroblast affinity whose membrane composition, modified by introduction of metabolic ligands, leads to a modulation of the synthesis activity of the targeted cells.  3 - Drug compositions according to claim 2 characterized in that the metabolic ligands are selected from initiators and modulators of cell synthesis with function or lipophilic radical by acylation.  4 - Drug compositions according to claim 2 and claim 3, characterized in that the initiators of cell synthesis are selected from fatty esters of ascorbic acid and in particular its palmitate.  5 - Drug compositions according to claim 2 and claim 3, characterized in that the cell synthesis modulator is collagen hydrolyzate palmitate.  6 - Drug compositions according to claim 2 and claim 3, characterized in that the modulators of cell synthesis are selected from the fatty esters of hydroxyproline and in particular its palmitate.  7 - Drug compositions according to any one of the preceding claims, characterized in that the vesicular vectors are chosen from vesicles, capsules or artificial spheres with a diameter which can range from 3 to 1000 nanometers, used to incorporate a substance. active either in an aqueous medium filling the interior volume of the vector or in the same wall of said vector and then to transport this active substance to a site where it will be released and used for different purposes.  8- Application of the drug compositions according to one of the preceding claims to increase the synthesis of collagenases to resorb dense fibrous tissue collagen I or keloids.  9 - Application of the drug compositions according to one of the preceding claims to the orientation of the synthesis of collagen to a particular structure such as collagens I and III.