CONTROLLED RELEASE SYSTEM OF PHYTOCANNABINOIDS FORMULATIONS SOLUBLE IN AQUEOUS MEDIA, METHODS AND USES THEREOF

Abstract

The present invention refers to a controlled release system of phytocannabinoids formulation soluble in aqueous media comprising phytocannabinoids, or derivatives thereof, a non-ionic surfactant and a cross-linked polymer, wherein the pH of the compositions ranges between 4 and 9, and wherein the weight ratio of cannabinoid/hyaluronic acid is 50:1 to 1:50, and the weight ratio cannabinoid/non-ionic surfactant is 1:30 to 1:1. The invention also refers to methods of producing the controlled release system of cannabinoids formulation soluble in aqueous medium and the pharmaceutical, cosmetic and nutraceutical applications thereof. The methods of the present invention allow the cross-linking of hyaluronic acid containing encapsulated or vehiculized phytocannabinoids at neutral pH, avoiding the degradation of the phytocannabinoids and preserving its functionality.

Claims

1. A multimatrix controlled release system of phytocannabinoids formulation soluble in aqueous media comprising a cross-linked polymer net which contains phytocannabinoids vehiculized through non-iconic surfactants, wherein the phytocannabinoids are selected from a phytocannabinoid, or a derivative thereof, or a mixture of phytocannabinoids, or derivatives thereof, wherein the polymer is selected from hyaluronic acid or derivative thereof, wherein the pH of the compositions ranges between 6 and 8, wherein the weight ratio of phytocannabinoid/polymer is 50:1 to 1:50, and wherein the weight ratio phytocannabinoid/non-ionic surfactant is 1:30 to 1:1.

2. A multimatrix controlled release system according to claim 1 wherein the phytocannabinoids are selected from cannabidiol, cannabigerol or a mixture thereof.

3. A multimatrix controlled release system according to claim 1 wherein the non-ionic surfactant is at least one poly (ethylene glycol)-block-poly (propylene glycol)-block-poly(ethyleneglycol) derivative.

4. A multimatrix controlled release system according to claim 3 wherein the poly (ethylene glycol)-block-poly (propylene glycol)-block-poly(ethyleneglycol) derivative is defined according to the formula: ##STR00004## wherein a is an integer of from 10 to 150 and b is an integer of from 15 to 65.

5. A multimatrix controlled release system according to claim 4 wherein the non-ionic surfactant comprises two poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol) derivatives.

6. A multimatrix controlled release system according to claim 5 wherein for one derivative a is 80 and b is 27 and for the other derivative a is 141 and b is 44.

7. A multimatrix controlled release system according to claim 5 wherein for one derivative a is 64 and b is 34, and for the other derivative a is 12 and b is 20.

8. A multimatrix controlled release system according to claim 1 wherein the phytocannabinoid is a cannabis sativa extract.

9. A multimatrix controlled release system according to claim 8 wherein the non-ionic surfactant is a combination of glyceryl citrate/lactate/linoleate/oleate and polyglyceryl-2 oleate.

10. (canceled)

11. A multimatrix controlled release system according to claim 1, wherein the polymer is a hyaluronic acid salt.

12. A multimatrix controlled release system according to claim 11, wherein the hyaluronic acid salt is of low molecular weight (M), where M≤0.75.Math.10.sup.6 Da, or a hyaluronic acid salt of high molecular weight (M), where M≤2.2.Math.10.sup.6 Da, or a mixture thereof.

13. A multimatrix controlled release system according to claim 12 wherein the hyaluronic acid salt is of low molecular weight, where 0.5.Math.10.sup.6 Da≤M≤0.75.Math.10.sup.6 Da or a hyaluronic acid salt of high molecular weight (M), where 1.9.Math.10.sup.6≤M≤2.2.Math.10.sup.6 Da or a mixture thereof.

14. A multimatrix controlled release system according to claim 11 wherein the hyaluronic acid salt is sodium hyaluronate.

15. A method of producing a multimatrix controlled release system of phytocannabinoids formulation soluble in aqueous media according to claim 1 comprising the steps of: a) Obtaining a solution by mixing hyaluronic acid, or a derivative thereof, previously dissolved in an aqueous solution, and a non-ionic surfactant, b) Obtaining a solution of a phytocannabinoid, or a derivative thereof, or a mixture of phytocannabinoids, or derivatives thereof, by dissolving said phytocannabinoids in a proper solvent, c) Adding the solution obtained in b) to the solution obtained in a), and d) Cross-linking the resulting solution obtained in c) in the presence of a cross-linking agent.

16. The method according to claim 15 wherein the phytocannabinoids used in step b) are selected from cannabidiol, cannabigerol or a mixture thereof.

17. A method, according to claim 15, wherein the non-ionic surfactant used in step a) is at least one poly (ethylene glycol)-block-poly (propylene glycol)-block-poly(ethyleneglycol) derivative.

18. Method according to claim 17 wherein the poly (ethylene glycol)-block-poly (propylene glycol)-block-poly(ethyleneglycol) derivative is defined according to the formula: ##STR00005## wherein a is an integer of from 10 to 150 and b is an integer of from 15 to 65.

19. The method according to claim 18 wherein the non-ionic surfactant comprises two poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a derivatives.

20. The method according to claim 19 wherein for one derivative a is 80 and b is 27 and for the other derivative a is 141 and b is 44.

21. The method according to claim 19 wherein for one derivative a is 64 and b is 34, and for the other derivative a is 12 and b is 20.

22. A method of producing a multimatrix controlled release system of phytocannabinoids formulation soluble in aqueous media according to claim 1 comprising the steps of: i. Obtaining a solution by dissolving hyaluronic acid, or a derivative thereof, in an aqueous solution, ii. Obtaining a solution of a phytocannabinoid, or a derivative thereof, or a mixture of phytocannabinoids, or derivatives thereof, by dissolving said phytocannabinoids in a solution that contains a non-ionic surfactant and a proper solvent iii. Adding the solution obtained in ii) to the solution obtained in i), and iv. Cross-linking the resulting solution obtained in iii) in the presence of a cross-linking agent.

23. The method according to claim 22 wherein a cannabis sativa extract containing phytocannabinoids is used in step i).

24. The method according to claim 23 wherein the non-ionic surfactant used in step ii) is glyceryl citrate/lactate/linoleate/oleate and polyglyceryl-2 oleate.

25. Method according to claim 15 further comprising a pH adjusting step after the cross-linking step, wherein when pH reached after the crosslinking step is acidic, the pH adjusting step proceeds by addition of a 0.25 M solution of sodium hydroxide (NaOH) until the required pH is reached, or when pH reached in step d) is basic, the pH adjusting step proceeds by addition of a 0.25 M solution of chlorhydric acid (HCl) until the required pH is reached.

26. (canceled)

27. Method according to claim 15 wherein the polymer is a hyaluronic acid salt of low molecular weight (M), where M≤0.75.Math.10.sup.6 Da, or a hyaluronic acid salt of high molecular weight (M), where M≤2.2.Math.10.sup.6 Da, or a mixture thereof.

28. Method according to claim 27 wherein the polymer is a hyaluronic acid salt of low molecular weight, where 0.5.Math.10.sup.6 Da≤M≤0.75.Math.10.sup.6 Da or a hyaluronic acid salt of high molecular weight (M), where 1.9.Math.10.sup.6≤M≤2.2.Math.10.sup.6 Da or a mixture thereof.

29. Method according to claim 27 wherein the hyaluronic acid salt is sodium hyaluronate.

30. Method according to claim 29 wherein the polymer solution comprises low molecular weight and/or high molecular weight sodium hyaluronate in a concentration between 0.1 and 3% (w/w), and preferably in a concentration between 0.5% and 2.5% (w/w) and most preferably in a concentration between 0.75 and 1.5% (w/w).

31. The method according to claim 15 wherein the aqueous solution medium used in step a) or i) is selected from: water, a buffer consisting of NaCl, in a concentration between 0.2 to 8%, and Na.sub.2HPO.sub.4 12H.sub.2O, in a concentration between 0.01% to 10% and NaH.sub.2PO.sub.4 2H.sub.2O, in a concentration between 0.001% and 10%, or a buffer consisting of sodium citrate dehydrate or a sodium citrate derivative, in a concentration between 0.002 to 2.5%, and citric acid (hydrated or dehydrated), in a concentration between 0.002% and 2%, and a buffer consisting of acetic acid in a concentration between 0.0005% and 0.1% and sodium acetate derivative in a concentration between 0.005% and 0.5%.

32. The method according to claim 15 wherein the solvent used to dissolve the cannabinoid is defined by the formula: ##STR00006## wherein R1-R4 are selected independently from H, OH, CH.sub.2OH, CH.sub.3, CH.sub.2CH.sub.3, C(O)CH.sub.3, C(O)OCH.sub.2CH.sub.3, CH.sub.2C(O)CH.sub.2CH.sub.3.

33. The method according to claim 32 wherein the solvent is selected from the group consisting of methanol, 1-propanol and isomers thereof, ethylene glycol, propylene glycol and mixtures thereof.

34. The method according to claim 15 wherein the cross-linking agent is selected from a carbodiimide derivative, a carbonyl imidazole derivative, a carbonyl benzotriazole derivative, a carbonyl triazole derivative or mixtures thereof.

35. The method according to claim 34 wherein an active ester forming molecules is present in the cross-linking step.

36. Method according to claim 34 wherein a dihydrazide derivative is present in the cross-linking step d).

37. Method according to claim 15 wherein, after the crosslinking step, a sterilization step is carried out.

38. Method, according to claim 37, wherein the sterilization process is performed by steam sterilization, in an autoclave at a temperature ranging from 120° C. to 140° C., for 15-20 minutes.

39. A pharmaceutical, cosmetic or nutraceutical composition comprising the multimatrix controlled release system of cannabinoids formulation according to claim 1.

40. A pharmaceutical composition, according to claim 39 for use in the treatment of inflammatory joint diseases.

41. A pharmaceutical composition according to claim 39 for use in the augmentation and/or repair of soft tissue and keratin materials.

42. A non-therapeutic use of the cosmetic or nutraceutical composition according to claim 39 for the relaxation, calming and moisturization of the skin and for the improvement of the well-being of the human body.

43. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0030] FIG. 1. Controlled release curve of phytocannabinoids. Concentration of CBD (mg/ml) vs time (h) for example 1 composition.

[0031] FIG. 2. Controlled release curve of phytocannabinoids. Concentration of CBG (mg/ml) vs time (h) for example 1 composition.

[0032] FIG. 3. Controlled release curve of phytocannabinoids. Concentration of CBG+CBD (mg/ml) vs time (h) for example 5 composition.

[0033] FIG. 4. Controlled release curve of phytocannabinoids. Concentration of CBG+CBD (mg/ml) vs time (h) for example 7 composition.

DETAILED DESCRIPTION OF THE INVENTION

[0034] In response to the needs of the state of the art, the authors of the invention have performed new methods for crosslinking polymers in the presence of phytocannabinoids vehiculized through non ionic surfactants. The subsequent product is sterilized, and it is suitable for pharmaceutical applications, for instance for the treatment of inflammatory joint diseases and for tissue filler applications. Non-sterilized product is suitable for cosmetic applications such as moisturizing gels, as well as for nutraceutical applications such as edible gels.

[0035] The obtained product is a multimatrix controlled release system of phytocannabinoids based on a cross-linked polymer net (matrix 1) which contains vehiculized phytocannabinoids through non-ionic surfactants (matrix 2). The fabrication of the whole system involves a chemical homogeneous synthesis.

[0036] Therefore, in a first aspect, the present invention refers to a controlled release system of phytocannabinoids formulation soluble in aqueous media comprising: [0037] a phytocannabinoid, or a derivative thereof, or a mixture of phytocannabinoids, or derivatives thereof, [0038] a non-ionic surfactant, and [0039] a cross-linked polymer,

[0040] wherein the pH of the compositions ranges between 4 and 9, preferably between 6 and 8, and wherein the weight ratio of phytocannabinoid/hyaluronic acid is 50:1 to 1:50, preferably 20:1 to 1:20, more preferably 10:1 to 1:10, and most preferably 2:1 to 1:2, and the weight ratio phytocannabinoid/non-ionic surfactant is 1:30 to 1:1, preferably 1:9 to 1:1.

[0041] Phytocannabinoids can be obtained not only from natural sources but also from chemical synthesis, biochemical synthesis or from genetically modified microorganism (R. K. Razdan, in The Total Synthesis of Natural Products, ed. J. ApSimon, 1981, vol. 4, pp. 185-262; U.S. Pat. No. 9,822,384B2: Production of cannabinoids in yeast). Accordingly, phytocannabinoids used in the formulation of the present invention can be natural or synthetic.

[0042] Phytocannabinoids can be selected, among others, from cannabigerolic acid, cannabigerolic acid monomethylether, cannabigerol, cannabigerol monomethylether, cannabigerovarinic acid, cannabigerovarin, cannabichromenic acid, cannabichromene, cannabichromevarinic acid, cannabichromevarin, cannabidiolic acid, cannabidiol, cannabidiol monomethylether, cannabidiol C4, cannabidivarinic acid, cannabidivarin, cannabidioreol, D9-(trans)-tetrahydrocannabinolic acid A, delta9-(trans)-tetrahydrocannabinolic acid B, D9-(trans)-tetrahydrocannabinol, D9-(trans)-tetrahydrocannabinolic acid C4, D9-(trans)-tetrahydrocannabinol-C4, D9-(trans)-tetrahydrocannabivarinic acid, D9-(trans)-tetrahydrocannabivarin, D9-(trans)-tetrahydrocannabiorcolic acid, D9-(trans)-tetrahydrocannabiorcol, D8-(trans)-tetrahydrocannabinolic acid, D8-(trans)-tetrahydrocannabinol, cannabicyclolic acid, cannabicyclol, cannabicyclovarin, cannabielsoic acid A, cannabielsoic acid B, cannabielsoin, cannabinolic acid, cannabinol, cannabinol methylether, cannabinol-C4, cannabivarin, cannabiorcol, cannabinodiol, cannabinodivarin, (−)-cannabitriol, (+)-cannabitriol, (±)-9,10-dihydroxy-D 6a(10a)-tetrahydrocannabinol, (−)-10-ethoxy-9-dihydroxy-D 6a(10a)-tetrahydrocannabinol, (±)-8,9-dihydroxy-D 6a(10a)-tetrahydrocannabinol, cannabidiolic acid tetrahydrocannabitriol ester and mixtures thereof.

[0043] In a particular embodiment, the phytocannabinoids are selected from cannabidiol, cannabigerol or a mixture thereof. In another particular embodiment, a cannabis sativa extract can be used as phytocannabinoids source. The cannabis sativa extract comes from any part of the cannabis sativa plant including flower, leaf, stem and seeds.

[0044] Phytocannabinoids are vehiculized in a non ionic surfactant. In order to aid the vehiculization of the phytocannabinoides, in a particular embodiment, non-ionic triblock copolymers derived of poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethyleneglycol) are used as surfactans. The poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethyleneglycol) are defined according to the formula:

##STR00001##

[0045] where a is an integer of from 10 to 150 and b is an integer of from 15 to 65.

[0046] In a particular embodiment, the formulation may comprise two poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a derivatives. When the formulation comprises two poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a derivatives, it is preferred that for one derivative a is 80, b is 27 and for the other derivative a is 141 and b is 44. Other known poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethyleneglycol) derivatives useful in the present invention are those where a is 64 and b is 34, a is 12 and b is 20.

[0047] In some particular embodiments, the combination of glyceryl citrate/lactate/linoleate/oleate and polyglyceryl-2 oleate can be used instead of the non-ionic triblock copolymers derived of poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethyleneglycol).

[0048] The polymer used in the formulations of the present invention can be natural or synthetic polymers. Examples of natural polymers are chondroitin sulfates, keratin sulfate, heparin and heparan sulfate, alginic acid and its biologically acceptable salts, starch, amylose, dextran, xanthan, pullulan, etc. Examples of synthetic polysaccharides are carboxy cellulose, carboxymethyl cellulose, alkyl celluloses such as hydroxyethyl cellulose and hydroxypropyl methyl cellulose (HPMC), oxidized starch etc.

[0049] In a preferred embodiment, the polymer is selected from chondroitin sulfate, alginic acid, or a derivative thereof, xanthan gum, carboxy cellulose, carboxymethyl cellulose sodium, hyaluronic acid or a derivative thereof. More preferably, the polymer to be crosslinked is a hyaluronic acid salt. In particular, it is selected from the sodium salt, the potassium salt and mixtures thereof.

[0050] In a preferred embodiment the hyaluronic acid salt is of low molecular weight (M), where M≤0.75.Math.10.sup.6 Da, or a hyaluronic acid salt of high molecular weight (M), where M≤2.2.Math.10.sup.6 Da, or a mixture thereof, more preferably the hyaluronic acid salt is of low molecular weight, where 0.5.Math.10.sup.6 Da≤M≤0.75.Math.10.sup.6 Da or a hyaluronic acid salt of high molecular weight (M), where 1.9.Math.10.sup.6≤M≤2.2.Math.10.sup.6 Da or a mixture thereof. Said low molecular or high molecular weight salts are of the same nature. In a most preferred embodiment, these salts consist of sodium hyaluronate.

[0051] The authors of the invention have performed new methods for obtaining the formulations of the invention, by crosslinking polymers in the presence of phytocannabinoids vehiculized through non ionic surfactants.

[0052] Therefore, in a second aspect, the present invention refers to a method of producing the controlled release system of phytocannabinoids formulation soluble in aqueous media comprising the steps of: [0053] a) Obtaining a solution by mixing a polymer, previously dissolved in an aqueous solution, and a non-ionic surfactant, [0054] b) Obtaining a solution of a phytocannabinoid, or a derivative thereof, or a mixture of phytocannabinoids, or derivatives thereof, by dissolving said phytocannabinoids in a proper solvent, [0055] c) Adding the solution obtained in b) to the solution obtained in a), and [0056] d) Cross-linking the resulting solution obtained in c) in the presence of a cross-linking agent.

[0057] In a particular embodiment, in step a), a solution that contains a polymer, preferably sodium hyaluronate or a derivative thereof, and a non-ionic surfactant, preferably poly(ethylene glycol)block-poly(propylene glycol)-block-poly(ethyleneglycol) derivative, is obtained. Upon addition of phytocannabinoids in step b), the non-ionic surfactant is able to form colloidal particles that contain phytocannabinoids.

[0058] Solution of step a) comprises low molecular weight and/or high molecular weight sodium hyaluronate in a concentration between 0.1 and 3% (w/w), and preferably in a concentration between 0.5% and 2.5% (w/w) and most preferably in a concentration between 0.75 and 1.5% (w/w), which is a sufficient amount of sodium hyaluronate to guarantee that the crosslinked hyaluronic acid containing phytocannabinoids has a homogeneous consistency.

[0059] The aqueous solution medium used in step a) is selected from: [0060] water, [0061] a buffer consisting of NaCl, in a concentration between 0.2 to 8%, and Na.sub.2HPO.sub.4 12H.sub.2O, in a concentration between 0.01% to 10% and NaH.sub.2PO.sub.4 2H.sub.2O, in a concentration between 0.001% and 10%, [0062] a buffer consisting of sodium citrate dehydrate or a sodium citrate derivative, in a concentration between 0.002 to 2.5%, and citric acid (hydrated or dehydrated), in a concentration between 0.002% and 2%, or [0063] a buffer consisting of acetic acid in a concentration between 0.0005% and 0.1% and sodium acetate derivative in a concentration between 0.005% and 0.5%.

[0064] In a preferred embodiment, the aqueous solution is a buffer consisting of 1.6% of NaCl, 0.12% of Na.sub.2HPO.sub.4 12H.sub.2O and 0.01% of NaH.sub.2PO.sub.4 2H.sub.2O.

[0065] In step b), a phytocannabinoid or a derivative thereof or a mixture of phytocannabinoids or derivatives thereof are dissolved in a proper solvent and added to the solution of step a). Preferably, the phytocannabinoids are selected from cannabidiol, cannabigerol or a mixture thereof. Under these conditions colloidal particles that contain cannabinoids are formed.

[0066] In step c), the cross-linking of the resulting solution is carried out in the presence of a cross-linking agent derived from a carbodiimide derivative, a carbonyl imidazole derivative, a carbonyl benzotriazole derivative, a carbonyl triazole derivative or mixtures thereof.

[0067] Optionally, an active ester forming molecule can be also added to the solution at the cross linking step, for example, N-hydroxysuccinimide (NHS), sulfo-N-hydroxysuccinimide (sulfo-NHS), hydroxybenzotriazole (HOBt), hexafluorophosphate benzotriazole tetramethyl uronium (HBTU) or 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, also called hexafluorophosphate azabenzotriazole tetramethyl uranium (HATU). Preferably, sulfo-NHS is used.

[0068] A dihydrazide derivative can also be added in the cross-linking step, for example, one selected from adipic acid dihydrazide, pimelic acid dihydrazide, malonic acid dihydrazide, ethylmalonic acid dihydrazide, carbonyl dihydrazide, oxalyldihydrazide or succinic dihydrazide.

[0069] The polymer to be crosslinked is preferably hyaluronic acid salt. It is more preferably selected from the sodium salt, the potassium salt and mixtures thereof, most preferably consists of the sodium salt (NaHA).

[0070] In a particular embodiment, the crosslinking process of the method of the invention is a process for crosslinking sodium hyaluronate and derivatives thereof in a solution that contains phytocannabinoids vehiculized in poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethyleneglycol) derivatives. The crosslinking process can be applied not only to sodium hyaluronate and derivatives thereof but also to other natural or synthetic polymers. Examples of natural polymers are chondroitin sulfates, keratin sulfate, heparin and heparan sulfate, alginic acid and its biologically acceptable salts, starch, amylose, dextran, xanthan, pullulan, etc. Examples of synthetic polysaccharides are carboxy cellulose, carboxymethyl cellulose, alkyl celluloses such as hydroxyethyl cellulose and hydroxypropyl methyl cellulose (HPMC), oxidized starch etc.

[0071] In the context of the cross-linking of this type of polymer (hyaluronic acid salt(s)) and of phytocannabinoids or mixtures thereof, in a preferred embodiment, the cross-linking reaction mixture contains: One hyaluronic acid salt of low molecular weight M, where M≤0.75.Math.10.sup.6 Da, preferably 0.5.Math.10.sup.6 Da≤M≤0.75.Math.10.sup.6 Da or one hyaluronic acid salt of high molecular weight M, where M≤2.2.Math.10.sup.6 Da, preferably 1.9.Math.10.sup.6≤M≤2.2.Math.10.sup.6 Da, or a mixture thereof. Said low molecular or high molecular weight salts are of the same nature and very advantageously consisting of sodium hyaluronate.

[0072] Sometimes, during the cross-linking process, the pH of the formulation may be outside the desired ranges (6 and 8), and then, a pH adjusting step after the cross-linking is necessary, wherein: [0073] If pH reached at the crosslinking is too acidic, the pH adjusting step proceeds by addition of a 0.25 M solution of sodium hydroxide (NaOH) until the required pH is reached, or [0074] If pH reached in step d) is basic, the pH adjusting step proceeds by addition of a 0.25 M solution of chlorhydric acid (HCl) until the required pH is reached.

[0075] In a particular embodiment, the solvent used to dissolve the phytcannabinoid in any of the method of the invention, is defined by the formula:

##STR00002##

[0076] wherein R1-R4 are selected independently from H, OH, CH.sub.2OH, CH.sub.3, CH.sub.2CH.sub.3, C(O)CH.sub.3, C(O)OCH.sub.2CH.sub.3, CH.sub.2C(O)CH.sub.2CH.sub.3.

[0077] The solvent is preferably selected from the group consisting of methanol, 1-propanol and isomers thereof, ethylene glycol, propylene glycol and mixtures thereof.

[0078] The crosslinking step involves the addition of a carbodiimide derivative. In a particular embodiment, the carbodiimide derivative is defined by the formula:

R.sup.1—N═C═N—R.sup.2

[0079] wherein R1 can be equal to R2. R1 and R2 are selected from cyclohexyl, isopropyl, 3-dimethylaminopropyl, ethyl, (2-morpholinoethyl). The carbodiimide can be as well in form hydrochloride or in form of methoxy-p-toluenesulfonate.

[0080] Instead of a carbodiimide derivative another crosslinking agents such as 1,1′-carbonyl diimidazole carbonyldibenzimidazole, carbonyldi-1,2,4-triazole, and carbonyldibenzotriazole may be used.

[0081] Crosslinking process could require the addition of an active ester forming molecule. Example of active ester forming molecule are N-hydroxysuccinimide (NHS), sulfo-N-hydroxysuccinimide (sulfo-NHS), hydroxybenzotriazole (HOBt), hexafluorophosphate benzotriazole tetramethyl uronium (HBTU), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate also called hexafluorophosphate azabenzotriazole tetramethyl uranium (HATU). In a preferred embodiment sulfo-N-hydroxysuccinimide (sulfo-NHS) is used.

[0082] Cross-linking is also performed by the addition of dihydrazide derivatives to the crosslinking mixture that contains carbodiimide derivatives as well as active ester forming molecules. Examples of dihydrazide derivatives are adipic acid dihydrazide, pimelic acid dihydrazide, malonic acid dihydrazide, ethylmalonic acid dihydrazide, carbonyl dihydrazide, oxalyldihydrazide, succinic dihydrazide.

[0083] Alternatively, the present invention contemplates another method of producing the controlled release system of phytocannabinoids formulation soluble in aqueous of the present invention comprising the steps of: [0084] i. Obtaining a solution by dissolving a polymer in an aqueous solution, [0085] ii. Obtaining a solution of a phytocannabinoid, or a derivative thereof, or a mixture of phytocannabinoids, or derivatives thereof, by dissolving said phytocannabinoids in a solution that contains a non-ionic surfactant and a proper solvent, [0086] iii. Adding the solution obtained in ii) to the solution obtained in i), and [0087] iv. Cross-linking the resulting solution obtained in iii) in the presence of a cross-linking agent.

[0088] Solution of step i) contains low molecular weight and/or high molecular weight sodium hyaluronate in a concentration between 0.1 and 3%, and preferably in a concentration between 0.5% and 2.5% and most preferably in a concentration between 0.75 and 1.5%, which is a sufficient amount of sodium hyaluronate to guarantee that the crosslinked hyaluronic acid containing phytocannabinoids has a homogeneous consistency.

[0089] The aqueous solution medium used in step i) is selected from: [0090] water, [0091] a buffer consisting of NaCl, in a concentration between 0.2 to 8%, and Na.sub.2HPO.sub.4 12H.sub.2O, in a concentration between 0.01% to 10% and NaH.sub.2PO.sub.4 2H.sub.2O, in a concentration between 0.001% and 10%, [0092] a buffer consisting of sodium citrate dehydrate or a sodium citrate derivative, in a concentration between 0.002 to 2.5%, and citric acid (hydrated or dehydrated), in a concentration between 0.002% and 2%, or [0093] a buffer consisting of acetic acid in a concentration between 0.0005% and 0.1% and sodium acetate derivative in a concentration between 0.005% and 0.5%.

[0094] In a preferred embodiment, the aqueous solution is a buffer consisting of 1.6% of NaCl, 0.12% of Na.sub.2HPO.sub.4 12H.sub.2O and 0.01% of NaH.sub.2PO.sub.4 2H.sub.2O.

[0095] In a particular embodiment, in step ii) a cannabis sativa extract containing phytocannabinoids is dissolved in a solution that contains a combination of glyceryl citrate/lactate/linoleate/oleate and polyglyceryl-2 oleate, as non-ionic surfactant, and a proper solvent, preferably propylenglycol. Under these conditions, nanocapsules which contain cannabinoids are formed.

[0096] Sometimes, during the cross-linking process, the pH of the formulation may be outside the desired ranges (6 and 8), and then, a pH adjusting step after the cross-linking is necessary, wherein [0097] If pH reached at the crosslinking is too acidic, the pH adjusting step proceeds by addition of a 0.25 M solution of sodium hydroxide (NaOH) until the required pH is reached, or [0098] If pH reached in step d) is basic, the pH adjusting step proceeds by addition of a 0.25 M solution of chlorhydric acid (HCl) until the required pH is reached.

[0099] In a particular embodiment, the solvent used to dissolve the phytcannabinoid in any of the method of the invention, is defined by the formula:

##STR00003##

[0100] wherein R1-R4 are selected independently from H, OH, CH.sub.2OH, CH.sub.3, CH.sub.2CH.sub.3, C(O)CH.sub.3, C(O)OCH.sub.2CH.sub.3, CH.sub.2C(O)CH.sub.2CH.sub.3.

[0101] The crosslinking step involves the addition of a carbodiimide derivative. In a particular embodiment, the carbodiimide derivative is defined by the formula:

R.sup.1—N═C═N—R.sup.2

[0102] wherein R1 can be equal to R2. R1 and R2 are selected from cyclohexyl, isopropyl, 3-dimethylaminopropyl, ethyl, (2-morpholinoethyl). The carbodiimide can be as well in form hydrochloride or in form of methoxy-p-toluenesulfonate.

[0103] Instead of a carbodiimide derivative another crosslinking agents such as 1,1′-carbonyl diimidazole carbonyldibenzimidazole, carbonyldi-1,2,4-triazole, and carbonyldibenzotriazole may be used.

[0104] Crosslinking process could require the addition of an active ester forming molecule. Example of active ester forming molecule are N-hydroxysuccinimide (NHS), sulfo-N-hydroxysuccinimide (sulfo-NHS), hydroxybenzotriazole (HOBt), hexafluorophosphate benzotriazole tetramethyl uronium (HBTU), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate also called hexafluorophosphate azabenzotriazole tetramethyl uranium (HATU). In a preferred embodiment sulfo-N-hydroxysuccinimide (sulfo-NHS) is used.

[0105] Cross-linking is also performed by the addition of dihydrazide derivatives to the crosslinking mixture that contains carbodiimide derivatives as well as active ester forming molecules. Examples of dihydrazide derivatives are adipic acid dihydrazide, pimelic acid dihydrazide, malonic acid dihydrazide, ethylmalonic acid dihydrazide, carbonyl dihydrazide, oxalyldihydrazide, succinic dihydrazide.

[0106] The resulting formulation, according to the methods of the invention, exhibit a concentration of hyaluronic acid between 0.1 and 3% (w/w), and preferably in a concentration between 0.5% and 2.5% (w/w) and most preferably in a concentration between 0.75 and 1.5% (w/w) and of cross-linking reagents in a concentration between 0.000025 M and 0.5 M and preferably between 0.05 M and 0.3 M and most preferably between 0.1 and 0.2 M. In a particular embodiment, for N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride the concentrations in weight are between 0.0004% and 8% (w/w), and preferably between 0.8% and 5% (w/w), and most preferably between 1.6 and 3.2% (w/w) with a pH between 4.5 and 8, which are compatible with an injectable use.

[0107] After the crosslinking step, the resulting formulation can be optionally sterilized, as in the case of injectable formulations. Sterilization process may be performed by steam sterilization, in an autoclave at a temperature ranging from 120° C. to 140° C. In particular, the sterilization can be performed at 121° for 15 to 20 minutes, preferably 15 min, to obtain F0>15 (sterilizing value). Dry-heat is also employed to achieve sterilization.

[0108] Sterilization may be performed by other means such as radiation sterilization including UV, X-rays, gamma ray, beta particles (electrons).

[0109] Sterilization may be also realized by chemical means including ethylene oxide, carbon-dioxide, ozone gas, hydrogen peroxide, nitrogen dioxide, glutaraldehyde and formaldehyde solutions, phthalaldehyde and peracetic acid.

[0110] The methods of the present invention afford scalability so that the fabrication method can be performed at industrial level (manufacturing). The method provides a net that includes homogeneously distributed colloid particles or nanocapsules and afford a controlled release system which can be optimized by: [0111] Hyaluronic acid with different molecular weight, [0112] Ratio of hyaluronic acid molecules with different molecular weight, [0113] Ratio hyaluronic acid/water, [0114] Cross-linking degree indicated by the cross-linking percentage, [0115] Cross-linking agent, [0116] % of colloid particles and nanocapsules, [0117] % of phytocannabinoids, [0118] Loading of colloid particles with different concentration (%) of phytocannabinoids, [0119] Type of phytocannabinoid and combination of them, [0120] Modulation of controlled release curves of phytocannabinoids, and [0121] Terminal sterilization.

[0122] The compositions of the invention are suitable for pharmaceutical, cosmetic and nutraceutical applications.

[0123] Therefore, in another aspect, the invention refers to a pharmaceutical composition comprising the phytocannabinoid formulations of the present invention and their uses in different pharmaceutical or medical applications. In particular, the present invention refers to the use of this pharmaceutical composition in the treatment of inflammatory joint diseases, taking advance of the viscosupplementation effect of the hyaluronic acid and the anti-inflammatory properties of hyaluronic acid and cannabinoids, especially cannabidiol.

[0124] The invention also refers to a pharmaceutical composition comprising the phytocannabinoid formulations of the present invention and its use in tissue filler applications. Specifically, the pharmaceutical composition of the invention affords sterile soft tissue filler compositions for the augmentation and/or repair of soft tissue and keratin materials, like the skin. These compositions can also comprise local anesthetics, such as lidocaine.

[0125] The present invention also refers to a cosmetic composition comprising the controlled release system of the present invention and its use for cosmetic applications. Specifically, the cosmetic composition of the invention affords compositions with relaxing, soothing and moisturizing effects on the skin.

[0126] Finally, the composition of the invention also refers to a nutraceutical composition comprising the controlled release system of the present invention and its use for nutraceutical applications. Specifically, nutraceutical compositions of the invention are useful for relaxation, calming and moisturization of the skin and for the improvement of the well-being of the human body.

[0127] The compositions of the invention can be administered by topical administration. Suitable topical compositions can be gels, ointments, creams, lotions, drops, etc. Topical compositions obtained by the methods of the invention do not need a sterilization step after the crosslinking step.

[0128] The composition of the invention can also be administered by systemic administration. This includes delivering the phytocannabinoid composition by injection, wherein the injection is intravenous, intra-articular, intramuscular, intradermal, intraspinal, intraperitoneal, subcutaneous, a bolus or a continuous administration.

[0129] The composition of the invention can also be administered by oral administration, such edible gels in the case of nutraceutical compositions.

[0130] The compositions of the invention can be administered including in a medical device. In an example, the composition can be drawn into a syringe for a water-based injection medium.

EXAMPLES

[0131] Analytical Techniques:

[0132] Reometry

[0133] The consistency of the gel is characterized at 25° C. by rheological measurement of the moduli of elasticity (G′) and viscosity (G″) as a function of the frequency (from 10 Hz to 0.01 Hz) using a controlled strain (1%) in AR 550 Rheometer (TA Instruments) and a cone-and-plate geometry of 40 mm diameter and a truncation (gap) of 115 μm.

[0134] High Performance Liquid Cromatography (HPLC)

[0135] This technique was used to determine the total content of phytocannabinoids (CBG/CBD) in the formulations and the amount of CBG/CBD encapsulated in the colloidal particles.

[0136] The analysis of the total amount of CBG/CBD in the system was performed by direct dilution of the samples in methanol followed by filtration through disposable 0.22 μm PVDF filters and subsequent injection in the chromatography equipment.

[0137] A HPLC-DAD analytical method according to Table 1 was developed in order to quantify the CBG and CBD concentration in the formulations. Such a method is common for both CBG and CBD.

TABLE-US-00001 TABLE 1 Chromatographic method for the quantification of CBG and CBD. System HPLC (1260 series Agilent Technologies) Column Zorbax Eclipse XCB-C18 (150 × 4.6 mm, 5 μm particle, Agilent) Mobile- Channel A: Ammonium formate 10 mM (pH 3.6, with Phase formic acid) Channel B: Acetonitrile Gradient 0-4 min 52-80% of B; 4-9.5 min 80% of B at 1 ml/min (post-run = 2 min) Detector DAD (210, 228 and 270 nm)

[0138] In this method a stock solution of 1000 mg/L of CBG/CBD in ethanol was prepared from which 1, 2, 3, 4, 5 and 6 mg/L standard dilutions of CBG/CBD in ethanol were made.

[0139] Dynamic Light Scattering (DLS)

[0140] DLS measurements were performed by diluting 70 μL of the samples in 900 μL of water followed by analysis in the DLS equipment at 173° measurement angle. Atenuator value, measurement position and count number were employed as measurement quality indicators (table 2).

TABLE-US-00002 TABLE 2 DLS analysis method parameters for the characterization of vehiculized CBG. Dispersant Water Temperature 25° C. Viscosity 0.8872 cP RI 1.330 Dielectric constant 78.5 Fitting model Smoluchowski Equilibration time 120 s Cell time Capillary cell DTS1070 Measurement angle 173° Number of measurements per sample 3

Example 1

[0141] 1 g of low molecular weight hyaluronic acid (MW 0.5-0.75 MDa) was dissolved in 99 mL phosphate saline buffer (pH 7.4) for 12 hours. 0.270 g of poly(ethylene glycol)a-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=80, b=27 and 0.270 g poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=141 b=44 was added via spatula to 5.40 g of hyaluronic acid solution. The resulting solution was stirred mechanically until complete solution of the polymers was reached. Following a solution of phytocannabinoid (64 mg of CBD, or 64 mg of CBG or a mixture of 32 mg of CBG+32 mg of CBD) in 100 microL of an organic solvent (methanol, isopropanol or propyleneglycol) was added to the hyaluronic acid/poly(ethylene glycol)a-poly(propylene glycol)b-block-poly(ethyleneglycol)a solution. The resulting solution was stirred mechanically for 48 hours. To that solution 230 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 65 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached.

Example 2

[0142] The experiment of example 1 was repeated adding a new component: adipic dihydrazide to the cross-linking step. The protocol was modified as follows: 230 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 65 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically for 1 hour. Afterwards 262 mg of adipic dihydrazide were added and the resulting mixture was shaken until complete dissolution of the dihydrazide is reached. The resulting mixture was let to cross-link for 24 h.

Example 3

[0143] The experiment of example 1 was modified so that half amount of EDC*HCl and of sulfo-NHS were used for the cross-linking step. The protocol was adapted as follows:

[0144] 115 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 33 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached.

Example 4

[0145] 0.9 g of low molecular weight hyaluronic acid (MW 0.5-0.75.Math.10.sup.6 Da) and 0.1 g of high molecular weight hyaluronic acid (MW 1.9-2.2.Math.10.sup.6 Da) were dissolved in 99 mL phosphate saline buffer (pH 7.4) for 12 hours. 0.270 g of poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=80, b=27 and 0.270 g poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=141 b=44 was added via spatula to 5.40 g of hyaluronic acid solution. The resulting solution was stirred mechanically until complete solution of the polymers was reached. Following a solution of phytocannabinoid (96 mg of CBD, or 96 mg of CBG or a mixture of 48 mg of CBG+48 mg of CBD) in 150 microL of an organic solvent (methanol, isopropanol or propyleneglycol) was added to the hyaluronic acid/poly(ethylene glycol)a-poly(propylene glycol)b-block-poly(ethyleneglycol)a solution. The resulting solution was stirred mechanically for 48 hours. To that solution 230 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 65 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached.

Example 5

[0146] The experiment of example 4 was modified so that half amount of EDC*HCl and of sulfo-NHS were used for the cross-linking step. The protocol was adapted as follows:

[0147] 115 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 33 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached.

Example 6

[0148] 0.5 g of low molecular weight hyaluronic acid (MW 0.5-0.75.Math.10.sup.6 Da) and 0.5 g of high molecular weight hyaluronic acid (MW 1.9-2.2.Math.10.sup.6 Da) were dissolved in 99 mL phosphate saline buffer (pH 7.4) for 12 hours. 0.270 g of poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=80, b=27 and 0.270 g poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=141 b=44 was added via spatula to 5.40 g of hyaluronic acid solution. The resulting solution was stirred mechanically until complete solution of the polymers was reached. Following a solution of phytocannabinoid (96 mg of CBD, or 96 mg of CBG or a mixture of 48 mg of CBG+48 mg of CBD) in 150 microL of an organic solvent (methanol, isopropanol or propyleneglycol) was added to the hyaluronic acid/poly(ethylene glycol)a-poly(propylene glycol)b-block-poly(ethyleneglycol)a solution. The resulting solution was stirred mechanically for 48 hours. To that solution 230 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 65 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached.

Example 7

[0149] The experiment of example 6 was modified so that half amount of EDC*HCl and of sulfo-NHS were used for the cross-linking step. The protocol was adapted as follows:

[0150] 115 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 33 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached.

Example 8

[0151] 1 g of high molecular weight hyaluronic acid (MW 1.9-2.2.Math.10.sup.6 Da) were dissolved in 99 mL phosphate saline buffer (pH 7.4) for 12 hours. 0.270 g of poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=80, b=27 and 0.270 g poly(ethylene glycol)a-block-poly(propylene glycol)b-block-poly(ethyleneglycol)a where a=141 b=44 was added via spatula to 5.40 g of hyaluronic acid solution. The resulting solution was stirred mechanically until complete solution of the polymers was reached. Following a solution of phytocannabinoid (64 mg of CBD, or 64 mg of CBG or a mixture of 32 mg of CBG+32 mg of CBD) in 100 microL of an organic solvent (methanol, isopropanol or propyleneglycol) was added to the hyaluronic acid/poly(ethylene glycol)a-poly(propylene glycol)b-block-poly(ethyleneglycol)a solution. The resulting solution was stirred mechanically for 48 hours. To that solution 230 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 65 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached.

Example 9

[0152] 1 g of low molecular weight hyaluronic acid (MW 0.5-0.75 MDa) was dissolved in 10 mL phosphate saline buffer (pH 7.4) for 12 hours. Following 0.60 grams of Mc Beauty Science Nano CBD Capsules (5%) were added to 5.4 g of the hyaluronic acid solution. The resulting solution was stirred mechanically for 48 hours. To that solution 230 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 65 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically until the complete dissolution of the reactants was reached. The resulting mixture was let to cross-link for 24 h.

[0153] INCI Name: Water (and) Cannabis Sativa Flower/Leaf/Stem Extract (and) Propylene Glycol (and) Glyceryl Citrate/Lactate/Linoleate/Oleate (and) Polyglyceryl-2 Oleate

[0154] CAS No.: 7732-18-5, 57-55-6, 9174-23-9, 9007-48-1

[0155] Composition (acc. to FDA):

[0156] A: >50% Water

[0157] C: 10-25% Cannabis Sativa Flower/Leaf/Stem Extract

[0158] C: 10-25% Propylene Glycol

[0159] 4.8-5.2% Cannabidiol

[0160] E: 1-5% Glyceryl Citrate/Lactate/Linoleate/Oleate

[0161] E: 1-5% Polyglyceryl-2 Oleate

Example 10

[0162] The experiment of example 9 was repeated adding a new component: adipic dihydrazide to the cross-linking step. The protocol was modified as follows: 230 mg of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC*HCl) and 65 mg of N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) was added via spatula. The resulting mixture was shaken mechanically for 1 hour. Afterwards 262 mg of adipic dihydrazide were added and the resulting mixture was shaken until complete dissolution of the dihydrazide is reached. The resulting mixture was let to cross-link for 24 h.

Example 11

Steam Sterilization

[0163] 1.2 g of material from EXAMPLES 1-9 were loaded manually in a rubber capped injection barrel of a 1.5 mL glass body syringe. The piston rod is plugged into the loaded injection barrel and the resulting loaded syringe is submitted to steam sterilization at 121° C. for 15 minutes.

Example 12

Controlled Release Experiment

[0164] 2.59 grams of material containing CBD from example 1 were introduced in a membrane that allows passing of the vehiculized cannabinoid through it. Filled membrane was put into contact with 29.91 g of phosphate buffer saline. The resulting solution was called controlled released solution. At different times 1 mL of solution was removed from the controlled release solution and the content was analysed by HPLC. For every 1 mL of controlled release solution that was removed for HPLC analysis another 1 mL of phosphate buffer saline was added to the controlled release solution so that the volume stayed constant (FIG. 1). FIG. 1 shows the controlled release of colloidal particles that contain CBD in a phosphate buffer medium. The analyzed concentration of CBD is cumulative, which means that concentration refers to the actual concentration of CBD in the controlled release solution and reaches a maximum value of 847 mg/L after 120 h of release experiment. 95% of the release of CBD takes place in the first 24 h of the experiment.

Example 13

Controlled Release Experiment

[0165] 2.45 grams of material containing CBG from example 1 were introduced in a membrane that allows passing of the vehiculized cannabinoid through it. Filled membrane was put into contact with 29.98 g of phosphate buffer saline. The resulting solution was called controlled released solution. At different times 1 mL of solution was removed from the controlled release solution and the content was analysed by HPLC. For every 1 mL of controlled release solution that was removed for HPLC analysis another 1 mL of phosphate buffer saline was added to the controlled release solution so that the volume stayed constant (FIG. 2). FIG. 2 shows the controlled release of colloidal particles that contain CBG in a phosphate buffer medium. The analyzed concentration of CBG is cumulative, which means that concentration refers to the actual concentration of CBG in the controlled release solution and reaches a maximum value of 680 mg/L after 120 h of release experiment. 96% of the release of CBD takes place in the first 24 h of the experiment.

Example 14

Controlled Release Experiment

[0166] 2.43 grams of material containing CBD and CBG from example 5 were introduced in a membrane that allows passing of the vehiculized cannabinoid through it. Filled membrane was put into contact with 28.82 g of phosphate buffer saline. The resulting solution was called controlled released solution. At different times 1 mL of solution was removed from the controlled release solution and the content was analysed by HPLC. For every 1 mL of controlled release solution that was removed for HPLC analysis another 1 mL of phosphate buffer saline was added to the controlled release solution so that the volume stayed constant (FIG. 3). FIG. 3 shows the controlled release of colloidal particles that contain a mixture of CBG+CBD in a phosphate buffer medium. The analyzed concentration of cannabinoids is cumulative, which means that concentration refers to the actual concentration of cannabinoids in the controlled release solution and reaches a maximum value of 235 mg/L after 6 h of release experiment. 98% of the maximum release of CBD takes place after 2 h of the release experiment.

Example 15

Controlled Release Experiment

[0167] 2.68 grams of material containing CBD and CBG from example 7 were introduced in a membrane that allows passing of the vehiculized cannabinoid through it. Filled membrane was put into contact with 29.33 g of phosphate buffer saline. The resulting solution was called controlled released solution. At different times 1 mL of solution was removed from the controlled release solution and the content was analysed by HPLC. For every 1 mL of controlled release solution that was removed for HPLC analysis another 1 mL of phosphate buffer saline was added to the controlled release solution so that the volume stayed constant (FIG. 4). FIG. 4 shows the controlled release of colloidal particles that contain a mixture of CBG+CBD in a phosphate buffer medium. The analyzed concentration of cannabinoids is cumulative, which means that concentration refers to the actual concentration of cannabinoids in the controlled release solution and reaches a maximum value of 255 mg/L after 24 h of release experiment. 79% of the maximum release of CBD takes place after 6 h of the release experiment.

Example 16

Physicochemical and Rheological Characterization of Sterilized Formulations

[0168]

TABLE-US-00003 Size of Zeta Cannabinoid particles potential Composition % (HPLC) Cross-linking pH (nm) PDI (mV) G′ G″ 1 CBG 0.67% Method 7.3 46.3 0.475 −33.6 28.2 5.9 example 1 2 CBD 0.35% Method 7.3 44.5 0.667 −8.0 26.9 6.0 example 1 3 CBG 0.39% Method 6.3 45.5 0.576 −7.8 3.3 2.6 example 2 4 CBD 0.35% Method 6.3 57.8 0.561 −15.1 45.0 5.6 example 2 5 CBG 0.60% Method 7.2 57.5 0.346 −21.3 65.6 8.6 example 3 6 CBD 0.41% Method 7.4 57.8 0.285 −29.4 38.9 8.1 example 3 7 CBD 0.55% Method 6.0 47.2 0.344 −45.1 31.9 12.0 CBG 0.58% example 4 8 CBD 0.58% Method 7.0 39.0 0.383 −16.8 46.3 10.4 CBG 0.61% example 5 9 CBD: 0.38% Method 6.0 72.2 0.564 −47.2 19.9 11.0 CBG: 0.38% example 6 10 CBD: 0.59% Method 7.1 60.3 0.761 −18.9 50.2 12.7 CBG: 0.56% example 7 11 CBG 0.99% Method 7.5 309.1 0.488 −18.4 77.0 13.5 example 8 12 CBD 0.79% Method 7.5 60.8 0.297 −25.1 20.5 5.7 example 8 13 CBD 0.17% Method 7.2 69.4 1.0 −17.8 15.4 3.8 example 9 [0169] Data from example 16 show that: Cannabinoids or mixtures of them are incorporated into colloidal particles based on non-ionic surfactants. Such particles are embedded in a matrix of crosslinked hyaluronic acid derivatives. [0170] The size of the colloidal particles is well defined and display polydispersity indexes (PDIs) down to 0.285. Such PDI values are typical for populations of particles with homogenous size. [0171] The zeta potential data with absolute values up to 47 mV indicates that the colloidal particles, which contain cannabinoids, are very stable and with low tendency to aggregation. [0172] G′ (elastic modulus) is larger than G″ (viscous modulus) which indicates that the elastic behaviour of the composition predominates over the viscous behaviour and demonstrates that the hyaluronic acid derivatives are chemically crosslinked (Stefano Santoro, Luisa Russo, Vincenzo Argenzio, Assunta Borzacchiello. Rheological properties of cross-linked hyaluronic acid dermal fillers. J. Appl Biomater Biomech 2011; Vol. 9 no. 2, 127-136). [0173] The use of lower amounts of crosslinking agents afford formulation with higher G′ and G″ values (example 3 vs example 1; example 5 vs example 4; example 7 vs example 6).