COMPOSITION COMPRISING AT LEAST ONE TRITERPENE AND/OR AT LEAST ONE TRITERPENOID AND/OR AT LEAST ONE OF THE GLYCOSYLATED FORMS THEREOF

Abstract

The present invention relates to a composition in the form of a thermoformed extrudate comprising at least one triterpene and/or at least one triterpenoid and/or at least one of the glycosylated forms thereof and at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof, said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof comprising at least one first amorphous phase and optionally one second crystalline phase. The present invention also relates to the method for manufacturing by thermoforming such a composition and to the use thereof.

Claims

1. A composition in the form of a thermoformed extrudate comprising at least one triterpene and/or at least one triterpenoid and/or at least one of the glycosylated forms thereof and at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof, said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof comprising at least one first amorphous phase and optionally one second crystalline phase.

2. The composition according to claim 1, characterized in that said thermoformed extrudate comprises a thermoformed mixture of said at least one triterpene and/or of said at least one triterpenoid and/or of said at least one of the glycosylated forms thereof and of said at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof.

3. The composition according to claim 1, characterized in that said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof predominantly comprises at least one first amorphous phase.

4. The composition according to claim 3, characterized in that said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof comprises between 51 and 100% by mass of an amorphous phase and between 0 and 49% by mass of a crystalline phase.

5. The composition according to claim 1, characterized in that said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof is tetracyclic, such as for example oleandrine, euphol or cucurbitacin, or pentacyclic, such as for example betulinic acid, oleanolic acid, boswellic acid, ursolic acid, lupeol, asiatic acid, madecassic acid, maslinic acid, jujubogenin or pseudojujubogenin.

6. The composition according to claim 5, characterized in that said boswellic acid is selected from the group consisting of α- and β-boswellic acids, for example α-boswellic acid, acetyl-α-boswellic acid, β-boswellic acid, acetyl-β-boswellic acid, 9,11-dehydro-α-boswellic acid, acetyl-9,11-dehydro-α-boswellic acid, 9,11-dehydro-β-boswellic acid, acetyl-9,11-dehydro-β-boswellic acid, 11-keto-β-boswellic acid, 11-keto-α-boswellic acid, 3-acetyl-11-keto-α-boswellic acid and 3-acetyl-11-keto-β-boswellic acid.

7. The composition according to claim 1, characterized in that said natural or synthetic proteins are selected from the group consisting of glycoproteins, collagens and/or collagen hydrolysates, plant proteins, animal proteins, the derivatives thereof and the mixtures thereof.

8. The composition according to claim 7, characterized in that said collagens and/or said collagen hydrolysates have a molecular weight comprised between 50 and 300000 Da, preferably between 100 and 275000 Da, preferably between 150 and 250000 Da, preferably between 200 and 225000 Da, preferably between 250 and 200000 Da, preferably between 300 and 175000 Da, preferably between 350 and 150000 Da, preferably between 400 and 125000 Da, preferably between 450 and 100000 Da, preferably between 500 and 75000 Da, preferably between 550 and 50000 Da, preferably between 600 and 40000 Da, preferably between 650 and 30000 Da, preferably between 700 and 20000 Da, preferably between 750 and 10000 Da, preferably between 800 and 9000 Da, preferably between 850 and 8000 Da, preferably between 900 and 7000 Da, preferably between 950 and 6000 Da, preferably between 1000 and 5000 Da, preferably between 1050 and 4000 Da, preferably between 1100 and 3000 Da, preferably between 1150 and 2000 Da, preferably between 1200 and 1000 Da.

9. The composition according to claim 1, characterized in that said oligosaccharides are selected from the group consisting of cyclodextrin, raffinose, rhamminose, rhamnose, stachyose, verbascose, trehalose, lactose, lactulose, maltose, the derivatives thereof and the mixtures thereof.

10. The composition according to claim 1, characterized in that said natural or synthetic polysaccharides are selected from the group consisting of starches, fibers, celluloses, hemicelluloses, glycogen, β-glucan, inulin, amylopectin, amylose, dextrin, maltodextrin, isomaltose, xylan, pullulan, agar-agar, carrageenans, mannans, fucoidan, gums, chitosan, chitin, xanthan, levan, neoserine, hyaluronic acid, hyaluronates, chondroitin sulphate, dermatan sulphate, keratan sulphate, the derivatives thereof and the mixtures thereof.

11. The composition according to claim 1, further comprising at least one additional natural or synthetic polymer selected from the group consisting of polyvinyl acetate, polyvinylpyrrolidone (PVP), polyvinylpyrrolidone-co-vinyl acetate, polyethylene-co-vinyl acetate, polyvinyl acid co-methacrylic acetate, polyethylene oxide, polylactide-co-glycolide, polyvinyl alcohol, polycarbophil, polycaprolactone, carnauba wax, ethylene-vinyl copolymer, lecithin, castor oil, hydrogenated soybean oil, waxes, isomalt, the derivatives thereof and the mixtures thereof.

12. The composition according to claim 1, further comprising at least one plasticizer.

13. The composition according to claim 12, characterized in that said at least one plasticizer is selected from the group consisting of polyols, lipids, sucrose esters, water, triethyl citrate, polyethylene glycol, dibutyl sebate, butyl stearate, glycerol monostearate, diethyl phthalate, the derivatives thereof and the mixtures thereof.

14. The composition according to claim 1, further comprising at least one additive selected from the group consisting of lubricants, surfactants, antioxidants, chelants, the derivatives thereof and the mixtures thereof.

15. The composition according to claim 1, further comprising at least one first additional compound of polyphenol type selected from the group consisting of phenolic acids, stilbenes, phenolic alcohols, lignans, flavonoids, the derivatives thereof and the mixtures thereof.

16. The composition according to claim 1, characterized in that it is packaged in the form of pellets, flakes, granules, powders, effervescent or non-effervescent tablets, injectable or non-injectable solutions, suspensions, gels, ointments or even in any other suitable form allowing administration to an animal or a human being.

17. A manufacturing method, in particular a method for manufacturing by thermoforming, a composition in the form of a thermoformed extrudate according to claim 1, characterized in that it comprises the following steps: a) a step of simultaneous or delayed supplying at least one triterpene and/or at least one triterpenoid and/or at least one of the glycosylated forms thereof and at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof, to be fed into an extruder, b) a step of mixing, in said extruder, said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof and said at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof, to form a mixture, and c) a step of hot extruding said mixture obtained in step b) in said extruder to obtain a thermoformed extrudate in which said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof comprises/comprise at least one first amorphous phase and optionally one second crystalline phase.

18. The method according to claim 17, characterized in that it comprises a preliminary step of premixing said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof and said at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof, in such a way to form a premixture intended to be fed into the extruder.

19. The method according to claim 17, characterized in that said hot extrusion step is carried out at an extrusion temperature comprised between 20 and 300° C., preferably at a temperature comprised between 40 and 270° C., preferentially at a temperature comprised between 50 and 250° C., preferably at a temperature comprised between 60 and 230° C., more preferentially at a temperature comprised between 70 and 220° C., more preferentially at a temperature comprised between 80 and 200° C., more preferentially at a temperature comprised between 90 and 180° C., more preferentially at a temperature comprised between 100 and 170° C., more preferentially at a temperature comprised between 120 and 160° C., more preferentially at a temperature comprised between 125 and 150° C.

20. The method according to claim 17, characterized in that said hot extrusion step is carried out at a rotation speed of an extrusion screw comprised between 20 and 900 rpm, preferably comprised between 50 and 300 rpm, preferably comprised between 100 and 250 rpm, preferentially equal to 250 rpm, preferentially equal to 100 rpm.

21. The method according to claim 17, characterized in that it comprises an additional step of cooling at the outlet of the extruder.

22. The method according to claim 17, characterized in that it comprises an additional step of treating the thermoformed extrudate at the outlet of the extruder, for example cutting at a pelletizer and/or grinding said thermoformed extrudate.

23. The composition in the form of a thermoformed extrudate according to claim 1 for use in the preventive and/or curative treatment, in human and/or in animal, of pathologies related to inflammations, pathologies related to the premature aging of cells, pathologies related to the cardiovascular system, pathologies related to the blood system, pathologies related to the gastrointestinal system, pathologies related to the endocrine system, pathologies related to the immune system, pathologies related to the central nervous system, skin diseases, diseases due to the presence of microorganisms and cancers and in the preventive and/or curative treatment of diabetes.

24. The composition in the form of a thermoformed extrudate obtained according to the method according to claim 17, said composition comprising at least one triterpene and/or at least one triterpenoid and/or at least one of the glycosylated forms thereof such as active ingredient and at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof, said at least one triterpene and/or said at least one triterpenoid and/or said at least one of the glycosylated forms thereof comprising at least one first amorphous phase and optionally one second crystalline phase.

Description

[0109] Other characteristics, details and advantages of the invention will emerge from the examples given below, not by way of limitation and with reference to the appended figures.

[0110] FIG. 1 is a graph illustrating the rate of solubilization of boswellic acids over time for different examples of compositions, in particular for different examples of thermoformed compositions, according to the invention.

[0111] FIGS. 2 and 3 are graphs illustrating the rates of dispersion over time of examples of different compositions, in particular for examples of different thermoformed compositions, according to the invention.

[0112] FIG. 4 is a graph illustrating the rate of solubilization of bacosides over time for different examples of compositions, in particular for different examples of thermoformed compositions, according to the invention.

[0113] FIG. 5 is a graph illustrating the rate of solubilization of asiaticoside over time for different examples of compositions, in particular for different examples of thermoformed compositions, according to the invention.

[0114] FIG. 6 is a graph illustrating the rate of dispersion of ursolic acid over time for different examples of compositions, in particular for different examples of thermoformed compositions, according to the invention.

EXAMPLES

Example 1: Thermoforming Manufacturing Method of a Composition According to the Invention in the Form of a Thermoformed Extrudate

[0115] Thermoformed compositions according to the invention comprising at least one triterpene and/or at least one triterpenoid and/or at least one of the glycosylated forms thereof comprising at least one amorphous phase, such as those which are the subject of Example 2 below, were obtained according to the following method which is also the subject of the present invention: [0116] a) a step of premixing at least one triterpene and/or at least one triterpenoid and/or at least one of the glycosylated forms thereof in the crystalline state in powder form and at least one polymer selected from the group consisting of natural or synthetic proteins, natural or synthetic oligosaccharides, natural or synthetic polysaccharides, the derivatives thereof and the mixtures thereof; [0117] b) a step of supplying said premix formed in step a) to be fed into a Pharma 11 type extruder of Thermo-Fischer®; [0118] c) a step of mixing, in said extruder, said premixture to obtain a mixture; [0119] d) a step of hot extrusion thermoforming said mixture obtained in step c) in said extruder to obtain a thermoformed extrudate, the hot extrusion step being carried out at a rotation speed of a extrusion screw of 100 rpm and at a temperature comprised between 40° C. and 180° C.; [0120] e) a step of cooling at the outlet of the extruder said thermoformed extrudate obtained in step d); and [0121] f) a step of cutting/grinding, at a grinder, the cooled thermoformed extrudate obtained in step e) in such a way to obtain a homogeneous powder.

[0122] The hot extrusion temperature (thermoforming temperature) at which the hot extrusion step is carried out is determined by the type of constituents used, in particular according to the type of polymer and/or plasticizer used, which the person skilled in the art is able to determine. Moreover, a person skilled in the art, in particular according to the type of extruder employed and in accordance with the general principle of the hot extrusion (HME), is able to define possible temperature steps in different zones along the extrusion screw(s) such that a gradual increase in the temperature within the material transported by the extrusion screw(s), this in an advancing direction of the material within the extruder. Typically, between zones defined along the extrusion screws(s), temperature differences in the range of 0 to 40° C. are observed. For example, in the context of the present invention, the below-tested compositions were obtained in a Pharma 11 type extruder of Thermo-Fischer® having 9 temperature zones which are as follows in an advancing direction of the material moving at a speed of 100 rpm: zone 1 (feed zone of the extruder)=ambient temperature; zone 2=120° C.; zone 3=120° C.; zone 4=120° C.; zone 5=130° C.; zone 6=140° C.; zone 7=150° C.; zone 8=155° C.; zone 9 (die)=160° C.

Example 2: Solubility Test of Thermoformed Compositions According to the Invention Comprising an Extract of Boswellia serrata Standardized to 65% Boswellic Acids

[0123] Different thermoformed compositions, obtained according to the manufacturing method described in Example 1, were tested in terms of solubility of boswellic acids present in an extract of Boswellia serrata standardized to 65% boswellic acids. This solubility was measured over time starting from the thermoformed extrudates obtained according to the invention. As indicated above, the thermoformed extrudates are in the form of a homogeneous powder (ground material) in which the triterpene and/or the triterpenoid comprises at least one amorphous phase.

[0124] All the solubility tests were carried out with a vane dissolution apparatus, starting with approximately 2 g of thermoformed extrudate, at a temperature of 37° C. under stirring at 50 rpm in 450 ml of a 0.1N HCl dissolution medium. These solubility tests were carried out according to the recommendations of the pharmacopoeia Ph.Eur.9.0 (Recommendations on Dissolution Testing). At determined times (after 30 min and after 2 h), a sample of 1 ml of mixture was taken to carry out a solubility test.

[0125] In order to carry out the solubility tests, the tested sample was filtered through a filter (PET, pore size of 0.45 μm, Macherey Nagel) before HPLC analysis (Luna column 5 μm C18(2) 100 A. 100*3 mm (Phenomenex); mobile phase: 85% A: methanol and 15% B: water/acetonitrile (95:5) at pH 2.8; flow rate: 0.6 mL/min, loop: 10 μl, t°=40° C.; wavelengths: 210 nm and 247 nm).

[0126] In practice, the solubility of triterpenes and/or of triterpenoids, in particular the solubility of boswellic acids contained in an extract of Boswellia serrata standardized to 65% boswellic acids, was evaluated by HPLC assay of the 6 main boswellic acids (α-boswellic acid, β-boswellic acid, 11-keto-β-boswellic acid (KBA), acetyl-11-keto-β-boswellic acid (AKBA), acetyl α-boswellic acid, acetyl β-boswellic acid) present in this extract of Boswellia serrata.

[0127] The thermoformed compositions according to the invention listed in Table 1 were formulated according to the method of the invention and tested in terms of solubility over time according to the principle indicated above (assay of the 6 main boswellic acids: the results presented are the solubility averages calculated by summing the solubilities of each of the 6 boswellic acids and dividing this sum by 6). A single extract of Boswellia serrata standardized to 65% boswellic acids in native crystalline form and in powder form (native BVV) was used as a control. The amounts mentioned in Table 1 are percentages by weight of the compounds used (subjected to the method according to the invention) based on the total weight of the composition.

TABLE-US-00001 TABLE 1 Extract of Boswellia serrata Glycerol (65%) (1) (2) Protein Compo 1 20 10 70 (3) Compo 2 20 10 70 (4) Compo 3 20 10 70 (5) (1) Dry extract of Boswellia serrata standardized to 65% boswellic acids (Vidya Herbs) (2) Glycerol (Sigma-Aldrich) (3) Fish collagen with a molecular weight of 3000 Da (Green Snow) (4) Hydrolyzed fish collagen with a molecular weight below 3000 Da (Kenney & Ross Ltd.) (5) Bovine gelatin with a molecular weight comprised between 1000 and 3000 Da (Lapi geltine S.P.A)

[0128] The results obtained are presented in FIG. 1. As can be seen, the natural proteins of collagen/gelatin type, used as a polymer, allow increasing the solubility of triterpenes and triterpenoids, in particular of boswellic acids. It should be noted that the solubility of boswellic acids varies according to the type/nature of collagen/gelatin but that this solubility is always increased compared to the control.

Example 3: Dispersion Test of Thermoformed Compositions According to the Invention Comprising an Extract of Boswellia serrata Standardized to 65% Boswellic Acids

[0129] Different thermoformed compositions, obtained according to the manufacturing method described in Example 1, were tested in terms of dispersion of the boswellic acids present in an extract of Boswellia serrata standardized to 65% boswellic acids. The dispersion was measured over time starting from the thermoformed extrudates obtained according to the invention. As indicated above, the thermoformed extrudates are in the form of a homogeneous powder (ground material) in which triterpene and/or triterpenoid comprises at least one amorphous phase.

[0130] All the dispersion tests were carried out with a vane dissolution apparatus starting with approximately 2 g of thermoformed extrudate, at a temperature of 37° C. under stirring at 50 rpm in 450 ml of a 0.1N HCl dissolution medium.

[0131] In order to carry out the dispersion tests, samples taken at determined times (after 30 min and after 2 h) were diluted in an appropriate solvent (mobile phase for HPLC) then filtered through a filter (PET, pore size of 0.45 μm, Macherey Nagel) before HPLC analysis (Luna column 5 μm C18(2) 100 A. 100*3 mm (Phenomenex); mobile phase: 85% A: methanol and 15% B: water/acetonitrile (95:5) at pH 2.8; flow rate: 0.6 mL/min, loop: 10 μl, t°=40° C.; wavelengths: 210 nm and 247 nm).

[0132] In practice, the dispersion of triterpenes and/or triterpenoids, in particular the dispersion of boswellic acids contained in an extract of Boswellia serrata standardized to 65% boswellic acids, was evaluated by HPLC assay of the 6 main boswellic acids (α-boswellic acid, β-boswellic acid, 11-keto-β-boswellic acid (KBA), acetyl-11-keto-β-boswellic acid (AKBA), acetyl α-boswellic acid, acetyl β-boswellic acid) present in this extract of Boswellia serrata.

[0133] The thermoformed compositions according to the invention listed in Table 2 were formulated according to the method of the invention and tested in terms of aqueous dispersion over time according to the principle indicated above (assay of the 6 main boswellic acids: the results presented are the dispersion averages calculated by summing the dispersions of each of the 6 boswellic acids and dividing this sum by 6). A single extract of Boswellia serrata standardized to 65% boswellic acids in native crystalline form and in powder form (native BVV) was used as a control. The amounts mentioned in Table 2 are percentages by weight of the compounds used (subjected to the method according to the invention) based on the total weight of the composition.

TABLE-US-00002 TABLE 2 Extract of Boswellia serrata Glycerol (65%) (1) (2) Protein Polysaccharide Compo 1 25 20 0   55 (3) Compo 2 25 20 70 (4) 0   Compo 3 25 20 10 (5) 45 (3) Compo 4 35 10 55 (6) 0   Compo 5 35 10 47 (6)  8 (3) (1) Dry extract of Boswellia serrata standardized to 65% boswellic acids (Vidya Herbs) (2) Glycerol (Sigma-Aldrich) (3) modified starch Cleargum CB90 (Roquette) (4) Rice protein (Green Snow) (5) Pumpkin seed protein (Green Snow) (6) 5000 Da-hydrolyzed collagen (Rousselot)

[0134] The obtained results are presented in FIG. 2 for compositions 1 to 3 and in FIG. 3 for compositions 4 and 5. As can be seen, each of the thermoformed compositions according to the invention gives rise to a percentage of dispersion significantly greater than that observed for the single extract of Boswellia serrata standardized to 65% boswellic acids in native crystalline form and in powder form (native BW).

Example 4: Solubility Test of Thermoformed Compositions According to the Invention Comprising an Extract of Bacopa monierii Standardized to 20% Bacosides

[0135] Different thermoformed compositions, obtained according to the manufacturing method described in Example 1, were tested in terms of solubility of bacosides present in an extract of Bacopa monierii standardized to 20% bacosides. This solubility was measured over time starting from the thermoformed extrudates obtained according to the invention. As indicated above, the thermoformed extrudates are in the form of a homogeneous powder (ground material) in which triterpene and/or triterpenoid comprises at least one amorphous phase.

[0136] All the solubility tests were carried out with a vane dissolution apparatus starting with approximately 4 g of thermoformed extrudate, at a temperature of 37° C. under stirring at 50 rpm in 900 ml of a 0.1N HCl dissolution medium. These solubility tests were carried out according to the recommendations of the pharmacopoeia Ph.Eur.9.0 (Recommendations on Dissolution Testing). At determined times (after 30 min and after 2 h), a sample of 1 ml of mixture was taken to carry out a solubility test.

[0137] In order to carry out the solubility tests, the tested sample was centrifuged (10000 rpm for 10 min at room temperature, Microstar 17 (VWR)), the supernatant was filtered through a filter (PET, pore size of 0.45 μm, Macherey Nagel) before HPLC analysis (C18 column, 5 μm 250*4.6 mm (Agilent); mobile phase: A: 0.1 mM phosphate buffer and B: Acetonitrile according to the following gradient:

TABLE-US-00003 TIME (MIN) A B 0 70% 30% 25 60% 40% 26 70% 30% 30 70% 30% Flow rate: 1.5 ml/min; loop, 20 μl; t°: 25° C.; wavelength 205 nm.

[0138] In practice, the solubility of triterpenes and/or of triterpenoids, in particular the solubility of bacosides contained in an extract of Bacopa monierii standardized to 20% bacosides, was evaluated by HPLC assay of the 5 main bacosides (Bacopaside I, Bacoside A3, Bacopaside II, Jujubogenin, Bacopasaponin C) present in this extract of Bacopa monierii.

[0139] The thermoformed compositions according to the invention listed in Table 3 were formulated according to the method of the invention and tested in terms of solubility over time according to the principle indicated above (assay of the 5 main bacosides: the results presented are the solubility averages calculated by summing the solubilities of each of the 5 bacosides and dividing this sum by 5). A single extract of Bacopa monierii standardized to 20% bacosides in native crystalline form and in powder form (native BC) was used as a control. The amounts mentioned in Table 3 are percentages by weight of the compounds used (subjected to the method according to the invention) based on the total weight of the composition.

TABLE-US-00004 TABLE 3 Extract of Bacopa Monierii Glycerol Protein Polysaccharide (20%) (1) (2) (3) (4) Compo 1 33 14 53 0 Compo 2 33 14 43 10 Compo 3 25 14 0 53 (1) Dry extract of Bacopa Monierii standardized to 20% bacosides (Vidya Herbs) (2) Glycerol (Sigma-Aldrich) (3) Hydrolyzed collagen with a molecular weight of 5000 Da (Rousselot) (4) Tackidex C760 (Roquette)

[0140] The results obtained are presented in FIG. 4. As can be seen, all the compositions according to the invention allow increasing the % of solubilized bacosides relative to the control.

Example 5: Solubility Test of Thermoformed Compositions According to the Invention Comprising Asiaticoside

[0141] Different thermoformed compositions, obtained according to the manufacturing method described in Example 1, were tested in terms of solubility of the asiaticoside. This solubility was measured over time starting from the thermoformed extrudates obtained according to the invention. As indicated above, the thermoformed extrudates are in the form of a homogeneous powder (ground material) in which triterpene and/or triterpenoid comprises at least one amorphous phase.

[0142] All the solubility tests were carried out with a vane dissolution apparatus starting with approximately 2 g of thermoformed extrudate, at a temperature of 37° C. under stirring at 50 rpm in 900 ml of a 0.1N HCl dissolution medium. These solubility tests were carried out according to the recommendations of the pharmacopoeia Ph.Eur.9.0 (Recommendations on Dissolution Testing). At determined times (after 30 min and after 2 h), a sample of 1 ml of mixture was taken to carry out a solubility test.

[0143] In order to carry out the solubility tests, the tested sample was centrifuged (10000 rpm for 10 min at room temperature, Microstar 17(VWR)), the supernatant was filtered through a filter (PET, pore size of 0.45 μm, Macherey Nagel) before HPLC analysis (C18 column, 5 μm 250*4.6 mm (Agilent); mobile phase: A: Water and B: Acetonitrile according to the following gradient:

TABLE-US-00005 TIME (MIN) A B 0 80% 20% 18 59% 41% 20 45% 55% 23 45% 55% 28 80% 20% 33 80% 20% Flow rate: 1 ml/min; loop, 10 μl; t°: 25° C.; wavelength 205 nm).

[0144] In practice, the solubility of triterpenes and/or triterpenoids, in particular the solubility of asiaticoside, was evaluated by HPLC assay.

[0145] The thermoformed compositions according to the invention listed in Table 4 were formulated according to the method of the invention and tested in terms of solubility over time according to the principle indicated above. Asiaticoside in native crystalline form and in powder form (native AS) was used as a control. The amounts mentioned in Table 4 are percentages by weight of the compounds used (subjected to the method according to the invention) based on the total weight of the composition.

TABLE-US-00006 TABLE 4 Asiaticoside Glycerol Protein Polysaccharide (1) (2) (3) (4) Compo 1 10 10 70 10 Compo 2 10 14 0 76 (1) Asiaticoside (FyzCo) (2) Glycerol (Sigma-Aldrich) (3) Hydrolyzed collagen with a molecular weight of 5000 Da (Rousselot) (4) Tackidex C760 (Roquette)

[0146] The results obtained are presented in FIG. 5. As can be seen, all the compositions according to the invention allow increasing the % of solubilized asiaticoside relative to the control.

Example 6: Dispersion Test of Thermoformed Compositions According to the Invention Comprising Ursolic Acid

[0147] Different thermoformed compositions, obtained according to the manufacturing method described in Example 1, were tested in terms of dispersion of ursolic acid. The dispersion was measured over time starting from the thermoformed extrudates obtained according to the invention. As indicated above, the thermoformed extrudates are in the form of a homogeneous powder (ground material) in which triterpene and/or triterpenoid comprises at least one amorphous phase.

[0148] All the dispersion tests were carried out with a vane dissolution apparatus starting with approximately 2 g of thermoformed extrudate, at a temperature of 37° C. under stirring at 50 rpm in 900 ml of an 0.1N HCl dissolution medium.

[0149] In order to carry out the dispersion tests, samples taken at determined times (after 30 min and after 2 h) were diluted in an appropriate solvent (mobile phase for HPLC) then filtered through a filter (PET, pore size of 0.45 μm, Macherey Nagel) before HPLC analysis (Luna column 5 μm C18(2) 100 A. 100*3 mm (Phenomenex); mobile phase Acetonitrile/Water/0.5% Ammonium acetate (67:12:21) flow rate: 1 mL/min, loop: 10 μl, t°=25° C.; wavelengths: 210 nm).

[0150] In practice, the dispersion of triterpenes and/or triterpenoids, in particular the dispersion of ursolic acid, was evaluated by HPLC assay.

[0151] The thermoformed compositions according to the invention listed in Table 5 were formulated according to the method of the invention and tested in terms of aqueous dispersion over time according to the principle indicated above. Ursolic acid in native crystalline form and in powder form (native AU) was used as a control. The amounts mentioned in Table 5 are percentages by weight of the compounds used (subjected to the method according to the invention) based on the total weight of the composition.

TABLE-US-00007 TABLE 5 Ursolic acid Glycerol Protein Polysaccharide (1) (2) (3) (4) Compo 1 10 10 80 0 Compo 2 10 14 66 10 Compo 3 10 14 0 76 (1) Ursolic acid (Fyzco) (2) Glycerol (Sigma-Aldrich) (3) Hydrolyzed collagen with a molecular weight of 5000 Da (Rousselot) (4) Tackidex C760 (Roquette)

[0152] The results obtained are presented in FIG. 6. As can be seen, all the compositions according to the invention allow increasing the % of dispersed ursolic acid relative to the control.

[0153] The present invention has been described in relation to the specific embodiments, which have a purely illustrative value and should not be considered as limitative. In general, it will appear obvious to the person skilled in the art that the present invention is not limited to the examples illustrated and/or described above.

[0154] The use of the verbs “comprise”, “include”, or any other variant, as well as the conjugations thereof, can in no way exclude the presence of elements other than those mentioned.

[0155] The use of the indefinite article “a”, “an”, or of the definite article “the”, to introduce an element does not exclude the presence of a plurality of these elements.