Dispersion composition containing hydrophobically modified hyaluronan derivative, zinc acetate, and curcumin, method of preparation and use thereof

Abstract

The invention relates to a long-term stable dispersion composition containing a hydrophobically modified hyaluronan derivative, zinc acetate, curcumin, and optionally other active substances and / or excipients. It also describes a method for its preparation and its use.

Description

[0001] Dispersion composition containing hydrophobically modified hyaluronan derivative, zinc acetate, and curcumin, method of preparation and use thereof.

[0002] Field of the invention

[0003] The present invention refers to a dispersion composition containing hydrophobically modified hyaluronan derivative, zinc acetate, curcumin and possibly other active substances and / or excipients. Furthermore, a method of its preparation and use is described.

[0004] Background of the invention

[0005] Carriers Based on Hyaluronic Acid

[0006] Acylated hyaluronan is known for its amphiphilic nature and its ability to form aggregates in aqueous environments (Ondreáš F., 2021; Eenschooten C., 2012). Using the solvent-exchange process, various water-insoluble or only sparingly soluble active substances, such as tocopherol, paclitaxel, phosphatidylcholine, or coenzyme Q10, can be encapsulated into it (CZ304654; Šmejkalová D., 2017; Šmejkalová D., 2014; Huerta-Angeles G., 2016). In

[0007]

[0008] addition to water, acylated hyaluronan can also be dissolved in physiological saline (0.9% NaCl). Using this approach, low amounts of curcumin were encapsulated: 1.1 to 3.2 wt% (relative to the carrier) into HA-C6 and 0.5 to 1.3 wt% into HA-C18:1 (Nešporová K., 2016).

[0009]

[0010] HA-C6 and HA-C18:1 with a molar mass of 15 x 103g mol-1were also further modified with Nile blue (DSNUC Blue < 1 %), where the curcumin binding capacity achieved by the solvent-exchange process into 1% HA-C6-Nile blue in phosphate buffer was low, 0.48 wt% relative to the carrier, and similarly low for 1% HA-C18:l-Nile blue in phosphate buffer, 0.62 wt% curcumin relative to the carrier (Starigazdová J., 2020). When studying the aggregation behavior of HA-C18:1 (Ondreáš F., 2021), the film-hydration technique, hydrating a film of the active substance after evaporation of the organic solvent, resulted in encapsulation of at most 0.8 wt% curcumin relative to the carrier. Long-term solution stability was not reported in these studies.

[0011] Curcumin can also be covalently bound to HA, although this may reduce its activity. A polycurcumin containing disulfide bonds was bound to 56 x 102g mol-1HA through amide bonds using a linker, resulting in a 25% curcumin-binding capacity. However, this concerns polycurcumin, not curcumin itself. The stability of curcumin in solution is not mentioned in this document (CN116731325A). HA with molar masses of 10 and 25 x 103g-mol'1was modified with curcumin by esterification through the carboxyl group, which led to micelle formation.However, within 8 hours after conjugation, curcumin degraded to 80 % of its initial amount (Fan Z., 2018), indicating insufficient stability. Esterification was also used to modify 4 x 103g mol-1HA with curcumin, resulting in a 27-fold increase in curcumin solubility in solution, corresponding to approximately 8.4 wt% curcumin relative to the carrier (HA). Nevertheless, the amount of curcumin within this material slightly decreased after 14 days at 4 °C (Hu J., 2018). This indicates that this covalent binding of curcumin to HA did not provide sufficient curcumin stability.

[0012] As a lipophilic compound, curcumin can be relatively well dissolved in oils. HA hydrophobically modified at the carboxyl group is capable of forming nanocapsules in the presence of oils. The amount of oil, and therefore the final amount of active substance in the nanocapsule formulation, is usually low, and long-term stability of active substances in these systems has not been demonstrated. Curcumin was also incorporated into nanocapsules by dissolving it in corn oil at a concentration of 15 mg / ml, and 3 pl of this oil was added to 1 ml of an aqueous solution of hydrophobically modified HA-C12 (1 mg / ml). The resulting curcumin concentration in solution was 45 pg / ml, binding efficiency approximately 4.5 wt% (relative to the carrier), which may be insufficient for certain applications. Long-term solution stability was not reported in this study (Czyzynska-Cichon I., 2021).

[0013] Curcumin Carriers Based on Other Systems in Combination with Hyaluronan

[0014] In hyalurosomes composed of hyaluronan (0.1-0.5 wt / vol% and 2 to 4 x 105g mol-1) and 180 mg / ml phospholipids, 10 mg / ml curcumin was encapsulated, corresponding to a binding capacity of approximately 5 to 6wt% (relative to the carrier). The hyalurosomes remained stable for 90 days at 25 °C with respect to the amount of encapsulated curcumin, which is not sufficiently long for common applications. It appears that hyaluronan does not form a direct surface layer, which may reduce the effectiveness of this composition; instead, through interactions with phospholipids, it forms a continuous polymer network (Manca. M..

[0015] 2015). Pluronic F127 was also used as a surfactant for curcumin encapsulation, with 57 x 104g mol-1HA employed to coat the particles. The encapsulated curcumin content was 3.3 ± 0.5 wt% (relative to the carrier), which is relatively low, and solution stability was not reported (Ji R, 2020). Ethosomes containing approximately 7.1 wt% curcumin relative to the carrier and composed of hydrogenated soybean phospholipids, DSPE-PEG2000, cholesterol, and 24 x 104g mol-1HA were stable for only 15 days at 5 °C (Zhang Y, 2019), which is highly insufficient.Hyaluronan with d-Block Elements

[0016] Coordination of transition-metal ions to HA, such as Ag+, Co2+, or Zn2+is well known. In particular, Zn2+coordinates at pH 6-6.5 (EP0413016B1). Coordination with zinc ions changed the conformation of the hyaluronan chain from a random coil (for the sodium ion) to a globular structure, thereby reducing the spatial dimensions of the molecule (Burger K., 2001).

[0017] Combination of Curcumin with d-Block Elements

[0018] One of the approaches for binding curcumin is its complexation with metals and other substances (Prasad S., 2022). Metal chelation occurs with the enolic form of curcumin through the -OH and =0 groups on the chain linking the two aryl rings (Banerjee S., 2015).

[0019] Curcumin can be encapsulated in high amounts into metal-organic frameworks (MOFs), which may also contain zinc and imidazole (ZIF). However, curcumin stability in ZIF is insufficient for applications, as it decreases significantly within 24 hours (Liu I, 2022). Through Zn2+, hyaluronan (HA) can also be coordinated to this system via COO⁻ groups (WO2022007153A1). Curcumin has also been encapsulated in high amounts into a system based on 5-aminolevulinic acid with Fe3+ions (CN114588271B). The metal-to-active-substance ratio in this system ranges from 1:0.7 to 1:200. The salt concentration ranges from 1 to 50 mM, and the active-substance concentration from 1 to 200 mM. The system can reportedly be stored at 4 °C either in solution or as a lyophilizate. The solution remains stable for 14 days at room temperature, which is insufficient for common applications. This system, however, does not contain hyaluronan.

[0020] Other Methods of Curcumin Encapsulation

[0021] The use of low-molecular- weight surfactants, such as Tween 80 in emulsions, increased the solubility of curcumin to 0.548 mg / ml, but even after surface modification with chitosan, low bioavailability remained an issue (Li J., 2016). In general, the stability of encapsulated curcumin tends to be problematic; for example, in copolymeric micelles, even retaining 56% of the initial curcumin content after three months is presented as a success (Leung H„ 2013), which is highly disadvantageous from a practical standpoint.

[0022] Effects of Curcumin

[0023] Curcumin, as a natural antioxidant, exhibits anti-inflammatory effects (Peng A, 2021). It has also demonstrated beneficial activity in wound healing (Castangia I., 2014). Curcumin showed positive effects in the prevention and treatment of skin disorders such as psoriasis andacne (Panahi Y., 2019). Favorable outcomes of topically or orally administered curcumin, particularly in encapsulated form, for selected skin diseases have been reported in preclinical and clinical studies and confirmed in a meta-analysis, which proposed curcumin as an alternative or complementary treatment for psoriasis (Zhang S., 2022). Positive effects of curcumin on undesirable oral symptoms in patients with oral submucous fibrosis have also been reported (Saso L, 2022). Clinical studies documented, and reviews and meta-analyses compared beneficial effects of using curcumin and turmeric formulations in preventing and reducing the severity, pain, and weight loss in patients with head and neck cancer undergoing radiotherapy or chemoradiotherapy (Wu C. F., 2024). Curcumin encapsulated in various formulations has also shown potential for topical administration in the treatment of ocular diseases such as dry eye syndrome or glaucoma, according to in vitro and in vivo studies (Davis B., 2018; Jahromy M. H., 2024 ). Curcumin-, turmeric-, and turmeric-extract-based systems are considered promising nutraceuticals due to their beneficial effects arising from the antioxidant and anti-inflammatory activity of curcumin (Kunnumakkara A., 2016). They may be used to improve physiological status not only in various diseases, such as chronic illnesses, cardiovascular disorders, inflammatory, metabolic, neurological, and dermatological conditions, and various infectious diseases. The anti-inflammatory and antioxidant properties of turmeric, curcumin, and its derivatives have also demonstrated strong potential for the treatment of inflammatory bowel diseases, such as ulcerative colitis and Crohn’s disease (Lin

[0024]

[0025] ¥,, 2022). However, the beneficial effects of curcumin are significantly limited by its low absorption and rapid elimination following oral administration. Therefore, enhanced formulations of curcumin are being developed to increase its bioavailability and thereby improve its therapeutic potential in pharmaceutical applications and dietary supplements (Corrêa Carvalho G., 2024).

[0026] Summary of the current state of the art

[0027] Curcumin is a natural active compound that has been bound into a wide range of materials. Only a limited number of carrier systems are capable of encapsulating a significant amount of curcumin. Systems that do offer this possibility, such as metal-organic frameworks, fail to ensure long-term stability of active substances in solution at laboratory temperature. Moreover, they cannot provide targeted delivery, controlled release, or the ability to overcome biological barriers, as they lack a stable HA-based shell that enables, among other things, targeting to various receptors such as CD44, TLR, or RHAMM (Lioan M., 2023), or improved penetration of substances into the skin ( Witting M., 2015). Nanoencapsulation is asuitable strategy for curcumin use due to increased solubility and bioavailability, the possibility of targeting, prolonged retention at the desired site, and protection against premature degradation (Hussain Z., 2017). Hydrophobically modified hyaluronic acid derivatives can bind various lipophilic active substances, but the encapsulated amounts are typically low, and these systems also struggle to maintain curcumin and other active substances stable in solution for extended periods at common temperatures (5-40 °C). Direct conjugation of curcumin to HA represents a complex preparation that may limit curcumin’s functionality, and it has not provided convincing evidence of good solution stability of curcumin.

[0028] Summary of the invention

[0029] The object of the invention is to prepare a composition containing curcumin in which the curcumin remains stable over a long period of time.

[0030] This object is achieved, and the above-mentioned disadvantages and shortcomings are eliminated, by the dispersion composition according to the present invention, characterised in that it contains at least one hydrophobically modified hyaluronan derivative of a general formula I serving as a carrier for encapsulation,

[0031] R1x

[0032]

[0033] where

[0034] R is H+or a physiologically acceptable metal cation selected from the group comprising any alkali-metal ions or alkaline-earth-metal ions or zinc ions, preferably Na+, K+, Mg2+, Ca2+, or Zn2+,

[0035] R1is -H or an acyl group -C(=O)CxHy, where x is an integer from 6 to 22 and j’ is an integer from 14 to 46, and CxHyis a linear or branched, saturated or unsaturated chain, wherein at least one repeating unit contains at least one acyl group,

[0036] with the proviso that

[0037] when R1is a Ce to Cio acyl group, n is an integer from 13 to 2500, preferably from 20 to 1500, with a degree of substitution of 5 to 70 %, preferably 8 to 60 %;

[0038] further with the proviso that, when R1is a Cn to Ci6 acyl group, n is an integer from 13 to 500 with a degree of substitution of 5 to 20 %, or n is an integer from 500 to1750 with a degree of substitution of 4 to 12 %;

[0039] further with proviso that, when R1is a C17 to C22 acyl group, n is an integer from 13 to 500, preferably from 20 to 250, with a degree of substitution of 3 to 15 %, preferably 4 to 10 %,

[0040] and wherein the concentration of the hydrophobically modified hyaluronan of general formula I is in the range of 0.03 to 0.4 wt% relative to the total weight of the composition.

[0041] and further comprises zinc acetate at a concentration in the range of 0.4 to 5 wt% relative to the total weight of the composition;

[0042] and further comprises encapsulated curcumin of a general formula II as a primary active substance.

[0043]

[0044] (II),

[0045] where R2is -H or -O-CH3, or a mixture of curcumins, wherein the curcumin is selected from the group comprising [(1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione], demethoxycurcumin, or bisdemethoxycurcumin;

[0046] wherein a concentration of curcumin of the general formula II or mixtures thereof is in the range of 0.003 to 0.1 wt% relative to the total weight of the composition;

[0047] and water or an aqueous salt solution selected from the group comprising aqueous solutions of chlorides or acetates of physiologically acceptable metals, wherein the physiologically acceptable metals are selected from the group comprising alkali-metal ions or alkaline-earth-metal ions or zinc ions, preferably Na+, K+, Mg2+, Ca2+, or Zn2+, more preferably an aqueous solution of zinc acetate or sodium chloride;

[0048] wherein the stability of the encapsulated curcumin in the dispersion composition is defined by maintaining at least 8 wt% of encapsulated curcumin relative to the weight of the carrier and at least 80 % of the initial amount of encapsulated curcumin for at least 3 months at 40 °C, or at least 90 % of the initial amount of encapsulated curcumin for at least 18 months at room temperature, or at least 90 % of the initial amount of encapsulated curcumin for at least 18 months at 5 °C.

[0049] According to a preferred embodiment of the composition of the present invention, R1is oleoyl, wherein the weight-average molar mass of oleoyl hyaluronan is in the range of 5 x 103to 3 x 104g mol-1, with a degree of substitution in the range of 3 to 15 %, preferably 1 x 104to 2 x 104g mol-1with a degree of substitution in the range of 5 to 10 %.

[0050] According to a further preferred embodiment of the present invention, the composition additionally comprises a secondary active substance or a combination thereof selected from the group of hydrophilic active substances comprising native hyaluronan, chondroitin sulfate, glucosamine sulfate, metformin, heparin, inositol, vitamin C and the group of B-vitamins, lactic acid, amino acids, a-hydroxy and P-hydroxy acids, peptides, water-soluble proteins, carboxymethyl glucan, schizophyllan, glucomannan, panthenol, urea, glycerol, pentylene glycol, preferably native hyaluronan, and / or from the group of hydrophobic active substances comprising dexamethasone, resveratrol, or cannabidiol, preferably dexamethasone, wherein the concentration of the secondary hydrophilic active substance or combinations thereof is 0.0001 to 3 wt%, and / or the concentration of the secondary hydrophobic active substance is 0.001 to 0.1 wt%.

[0051] Preferably, the native hyaluronan has a weight-average molar mass in the range of 5 x 103to 2 x 106g mol-1, more preferably in the range of 1 x 104to 1.5 x 106g mol-1.

[0052] According to additional another preferred embodiment of the present invention, the composition further comprises an excipient or a combination thereof selected from the group comprising viscosity regulators, preferably xanthan gum, guar gum, carboxymethylcellulose, sodium alginate, pectin, carbomer, polyethylene glycol, polyvinylpyrrolidone, hydrolyzed collagen, gelatin, more preferably xanthan gum; preservatives, preferably aromatic acids and their derivatives, more preferably benzoic acid, salicylic acid, dehydroacetic acid, potassium sorbate, parabens; alcohols, preferably ethanol, isopropanol, benzyl alcohol, phenoxyethanol, phenethyl alcohol, sorbitol, pentylene glycol; sugars, preferably sucrose, glucose, fructose, mannose, galactose; salts, preferably sodium chloride, wherein the concentration of the excipient or combinations thereof is 0.001 to 5 wt%.

[0053] According to another embodiment of the invention, the invention further comprises a method for preparing the composition according to the present invention, wherein curcumin dissolved in an organic solvent is added to an aqueous solution of at least one hydrophobically modified hyaluronan derivative of the general formula I and zinc acetate, after which the organic solvent is evaporated, and the non-encapsulated curcumin is subsequently removed, preferably by filtration, more preferably through a 1 μm glass filter, wherein the organic solvent is selected from the group consisting of acetonitrile, ethanol, isopropanol, preferably isopropanol.

[0054] Preferably, at least one secondary active substance and / or at least one excipient is further addedto the aqueous solution of at least one hydrophobically modified hyaluronan derivative of the general formula I serving as the encapsulation carrier and zinc acetate.

[0055] According to additional another preferred embodiment of the method of the present invention, the concentration of the hydrophobically modified hyaluronan is in the range of 0.3 to 4 g / 1, preferably 0.5 to 2 g / 1, more preferably 1 g / 1.

[0056] According to additional another preferred embodiment of the method of the present invention, the concentration of zinc acetate is in the range of 25 to 200 mM, preferably 30 to 70 mM, more preferably 55 mM.

[0057] According to additional another preferred embodiment of the method of the present invention, the concentration of curcumin in the organic solvent is in the range of 0.5 to 5 g / 1, preferably 2 to 4 g / 1.

[0058] According to additional another preferred embodiment of the method of the present invention, at least one secondary hydrophilic active substance dissolved in an aqueous solution is added, preferably in an aqueous solution of zinc acetate at a concentration of 30 to 70 mM.

[0059] According to yet another preferred embodiment of the method of the present invention, at least one hydrophobic active substance dissolved at a concentration of 0.5 to 5 g / 1, preferably 2 to 4 g / 1, in an organic solvent is added, after which the organic solvent is evaporated and the non-encapsulated hydrophobic active substance is removed, preferably by filtration, more preferably through a 1 μm glass filter.

[0060] According to additional another preferred embodiment of the method of the present invention, after evaporation of the organic solvent, the volume of the prepared dispersion composition is replenished with water or an aqueous salt solution selected from the group comprising aqueous solutions of chlorides or acetates of physiologically acceptable metals, wherein the physiologically acceptable metal ions are selected from the group comprising alkali-metal ions, alkaline-earth-metal ions, or zinc ions, preferably Na+, K+, Mg2+, Ca2+, or Zn2+, more preferably an aqueous solution of zinc acetate or sodium chloride.

[0061] According to additional another preferred embodiment of the method of the present invention, the composition is subsequently converted into a dry form, preferably by lyophilization.

[0062] A further embodiment of the invention comprises a pharmaceutical, cosmetic, or nutraceutical composition containing the dispersion composition according to the present invention.According to additional another preferred embodiment, the pharmaceutical, cosmetic, or nutraceutical composition according to the invention is in a form selected from the group comprising a cream, gel, emulsion, serum, or hydrogel.

[0063] Preferably, the composition is in the form of an emulsion, wherein the concentration of the dispersion composition is in the range of 0.001 to 99.999 wt / wt %, preferably 0.01 to 10 wt / wt %, more preferably 0.1 to 5 wt / wt %.

[0064] Preferably, the composition is in the form of a serum, wherein the concentration of the dispersion composition is in the range of 0.001 to 100 wt / wt %, preferably 0.01 to 10 wt / wt %, more preferably 0.1 to 5 wt / wt %.

[0065] Preferably, the composition is in the form of a cream, wherein the concentration of the dispersion composition is in the range of 0.001 to 50 wt / wt %, preferably 0.01 to 10 wt / wt %, more preferably 0.1 to 5 wt / wt %.

[0066] Preferably, the composition is in the form of a gel or hydrogel, wherein the concentration of the dispersion composition is in the range of 0.001 to 100 wt / wt %, preferably 0.01 to 95 wt / wt %, more preferably 0.1 to 10 wt / wt %.

[0067] According to additional another preferred embodiment, the pharmaceutical or cosmetic composition according to the present invention comprises the dispersion composition and at least one cosmetic or pharmaceutical excipient selected from the group comprising an oil, wax, butter, emulsifier, secondary active substance, viscosity regulator, and preservative.

[0068] According to a further embodiment, the dispersion composition according to the present invention, or the pharmaceutical or cosmetic composition according to the invention, is for use in a pharmaceutical application, preferably for treatment of the skin or mucous membranes, or for topical or ophthalmic administration.

[0069] According to additional another preferred embodiment, the nutraceutical composition according to the present invention comprises the dispersion composition and is preferably in a form selected from the group comprising a gel, emulsion, solution, syrup, jelly, or hydrogel.

[0070] According to additional another preferred embodiment, the dispersion composition according to the present invention, or the nutraceutical composition according to the present invention, is used for oral administration, preferably in the form of a dietary supplement.

[0071] According to additional another preferred embodiment, the dispersion composition according to the present invention, or the pharmaceutical or cosmetic composition according to the invention, is used for cosmetic application, preferably for cosmetic treatment of the skin.

[0072] Detailed Description of the InventionThe subject of the invention is a dispersion composition with particle sizes of 10 to 500 nm containing a hydrophobically modified hyaluronan derivative, preferably an acylated hyaluronan (A-HA) according to the general formula I, zinc acetate, and curcumin (CURC), prepared by the method of the invention. These dispersions are long-term stable across a wide temperature range and can contain, in high amounts, an additional non-covalently bound active substance.

[0073] The dispersion composition according to the present invention is intended for pharmaceutical, cosmetic, or nutraceutical purposes.

[0074] Preferably, it has the following composition, with the individual components expressed in weight percentages relative to the total weight of the composition: Water 86.4 to 99.567 wt%, zinc acetate 0.4 to 5 wt%, acylated hyaluronan 0.03 to 0.4 wt%, curcumin 0.003 to 0.1 wt%.

[0075] Preferably, the composition also contains the secondary hydrophilic active substance in an amount up to 3 wt%, a secondary hydrophobic active substance in an amount up to 0.1 wt%, an excipient in an amount up to 5 wt%, or mixtures thereof.

[0076] The dispersion composition according to the invention preferably comprises secondary active substances selected from the group comprising vitamins, pharmaceuticals, cytostatics, steroids, and cosmetically active substances, preferably resveratrol (RST), cannabidiol (CBD), dexamethasone (DXM), or native hyaluronan.

[0077] Excipients may be incorporated to improve the properties of the final dispersion composition or its solid form. The excipients may preferably be xanthan gum, sorbitol, sucrose, pentylene glycol, or other high-molecular-weight or low-molecular-weight substances.

[0078] During the preparation of the dispersion composition according to the present invention, a specific method of preparation and the composition must be followed in order to obtain the above-described dispersion composition of the invention. A solvent-exchange process is used. Other processes, such as direct mixing or formation of an active-substance film after evaporation of the organic solvent followed by hydration with a carrier solution, do not work (see Comparative Examples 66 and 67). Curcumin is dissolved in a suitable organic solvent. As the organic solvent, ethanol or isopropanol is preferably used, and acetonitrile may also be employed (Examples 1, 27, 28). When tetrahydrofuran or chloroform is used, an insufficient amount of curcumin is encapsulated in the dispersion composition (see Examples 69 and 70). When methanol or acetone is used, a sufficient amount of curcumin is encapsulated, but the curcumin in the dispersion composition is not sufficiently stable over time (see Examples 68 and 71). The concentration of curcumin or another secondary active substance in the organicsolvent is in the range of 0.5 to 5 mg / ml (Examples 1, 2, 23, 24, 25). Separately, the hydrophobically modified amphiphilic hyaluronan derivative, preferably a C6–C22acylated hyaluronan, is dissolved at concentrations of 0.3 to 4 mg / ml (Examples 4 and 44) in a zinc acetate solution with a concentration of 25 to 200 mM (Examples 32 and 35); otherwise, the described dispersion composition with the required properties does not form. Dissolution may be facilitated by supplying energy, for example by using ultrasound or heating to an elevated temperature. The volume ratio of the aqueous phase to the organic phase is preferably from 1: 1 to 10:1 (Examples 1, 23, 24, 25). The solutions are mixed under continuous stirring at temperatures between 15 and 40 °C. The organic solvent is evaporated at 34 °C while gradually reducing the pressure from atmospheric pressure to 45 x 102Pa. The volume or mass of the solution after evaporation may be adjusted to the desired value using water or another solution compatible with the prepared dispersion composition, for example aqueous solutions of metal acetates such as zinc acetate, so that the resulting concentration of zinc acetate is 0.4 to 5 wt % relative to the total weight of the composition of the invention, or aqueous solutions of metal chlorides such as NaCl, for example 0.9 wt% NaCl. The volume of the replenishing solution may be up to five times the original volume of the aqueous phase. The prepared solution is freed from non-encapsulated primary and, where applicable, secondary active substances, for example by centrifugation or filtration through a selected filter, such as a 1 pm filter made of a selected material, for example a glass filter. Additional substances, such as native hyaluronan or pentylene glycol, may be added to the prepared solution. The curcumin content, particle size, and other analyses were performed on the resulting dispersion composition according to the invention. The prepared dispersion, containing particles of a suitable size of 10 to 500 nm, contains a high amount of curcumin, above 10 wt% relative to the carrier (calculation described in the definitions of terms used in the Examples of the invention), and optionally additional active substances, and is long-term stable in solution.

[0079] Suitable physical properties of the dispersion composition according to the present invention:

[0080] Mean particle size (measured by DLS - dynamic light scattering) - in the range of 10 to 500 nm, preferably 50 to 300 nm. The measurement procedure and evaluation are described in Example 86.

[0081] Curcumin binding capacity relative to the carrier - at a carrier concentration of 0.03 to 0.4 wt %, the curcumin binding capacity is higher than 10 wt %, calculated relative to the mass of the carrier. The determination process is described in Example 87, and the process for determining the component ratio is described in Example 99.Stability of encapsulated curcumin in the dispersion composition according to the invention is characterized by the preservation of at least 8 wt % of encapsulated curcumin relative to the mass of the carrier, and at least 80 % of the original amount of encapsulated curcumin for at least 3 months at 40 °C, or at least 90 % of the original amount of encapsulated curcumin for at least 18 months at room temperature, or at least 90 % of the original amount of encapsulated curcumin for at least 18 months at 5 °C.

[0082] Binding capacity of secondary active substances - at a carrier concentration of 0.3 to 4 mg / ml, the binding capacity for an additional active substance is higher than 10 wt %, calculated relative to the mass of the carrier. The process for determining resveratrol, dexamethasone, and cannabidiol is described in Examples 88, 89, and 90.

[0083] Acylated hyaluronan (A-HA) may be hyaluronan acylated with substituents of chain length C6to C22(Examples 1 to 22). The substituent may be attached to A-HA, for example, by esterification, preferably using the mixed anhydride procedure, to substitute preferably the OH on C6 of HA while preserving the carboxyl groups of HA (Huerta-Ángeles G., 2014, CZ304654).

[0084] The substituent may also contain double bonds, such as C18:1or C22:1(Examples 1, 14, 15). The degree of substitution of A-HA is in the range of 1 to 70 %, defined as the number of acyl groups per 100 HA disaccharides and may be determined, for example, by ¹H NMR spectroscopy (DSNMR, Huerta-Ángeles G., 2014) or by gas chromatography (DSGC,

[0085]

[0086]

[0087] The weight-average molar mass (Mw) of A-HA is in the range of 5 x 103to 1.5 x 106g mol-1, determined by size-exclusion chromatography with multi-angle light scattering detection (SEC-MALS). With increasing HA chain Mw, increasing hydrophobic substituent length, and increasing DS, the solubility of A-HA decreases. To form a stable dispersion, the acylated hyaluronan must be well dissolved during the preparation process in zinc acetate. This means that for A-HA with shorter substituents below CIO, the dispersion compositions of the present invention with advantageous properties can be prepared using Mwfrom 5 x 103to 1 x 106g mol-1and degree of substitution of 5 to 70 % (Examples 11, 12, 13, 22). For A-HA with medium-length hydrophobic substituents C11-C16, the dispersion compositions of the present invention with advantageous properties can be prepared using Mwfrom 5 x 103to 2 x 105g mol-1with degree of substitution of 5 to 20 % (Examples 17, 62), and from 2 x 105to 7 x 105g mol-1with degree of substitution of 4 to 12 % (Examples 19, 20, 21, 46). For A-HA with longer hydrophobic substituents C17-C22, the dispersion compositions of the present invention with advantageous properties can be prepared using Mwfrom 5 x 103to 2 x 105g mol-1with degree of substitution of 3 to 11 % (Examples 14, 15, 16, 18). Theconcentration of A-HA must also be appropriately selected to ensure good solubility with respect to the type and degree of hydrophobic substitution and the polymer’s molar mass. Furthermore, to form a stable dispersion, the concentration of acylated hyaluronan must be higher than 0.2 mg / ml, ensuring sufficient material to coat curcumin and optionally other active substances, and stabilize the dispersion. The advantageous concentration range of A-HA is 0.3 to 4 mg / ml.

[0088] The method described in this invention, unlike the current state of the art, enables the preparation of the dispersion composition according to the present invention that contains a large amount of curcumin, long-term stable in solution, with enhanced bioavailability, effective penetration of biological barriers, and biological activity due to the selected HA derivatives, the combination with zinc acetate, and the specific preparation procedure. The combination with zinc also provides additional beneficial effects, such as antimicrobial activity (Prasad S, 2022). Furthermore, the dispersion composition described in this invention allows for the simultaneous encapsulation of additional secondary active substances in high amounts, which may lead to synergistic effects.

[0089] The presence of zinc acetate is essential for preparing the stable dispersion composition according to the present invention. Bulkier carboxylate anions, such as citrate or gluconate, do not support the formation of the stable dispersion (Examples 47 and 48). When using other zinc salts, such as sulfate or perchlorate, it is also not possible to achieve the desired high curcumin binding capacity (Examples 51 and 58). The zinc cation is crucial for stabilizing the dispersion, since systems containing acetates of s-block elements (sodium, magnesium, calcium acetates) and ytterbium acetate (f-block element) exhibited orders of magnitude lower curcumin binding (Examples 52, 53, 54, 61). Systems with acetates of d-block elements (cobalt(II), cadmium(II), manganese(II), copper(II) acetates) did achieve high curcumin binding, but they were not stable over time (Examples 56, 57, 59, 60). The concentration of zinc acetate must be greater than 20 mM, as lower concentrations may still provide high curcumin binding, but they are not sufficiently stable in solution (Examples 45 and 65).

[0090] In the above-described dispersion composition according to the present invention, it is possible, besides curcumin as the primary active substance, to encapsulate large amounts of certain secondary active substances, such as hydrophobic compounds, such as resveratrol, cannabidiol, or dexamethasone (Examples 36 to 40). These substances would not encapsulate into A-HA in comparable amounts on their own (Examples 74, 75, 76). The secondary hydrophobic active substance may be added either before the addition of curcumin in the organic solvent, or after the addition of curcumin in the organic solvent, followed byevaporation of the organic solvent for both active substances. Alternatively, curcumin may be added first, the organic solvent evaporated, then another hydrophobic active substance may be added in an organic solvent, and the solvent evaporated again. The key requirement is that curcumin must be present in the system containing A-HA and zinc acetate during the first evaporation step.

[0091] In the above described dispersion compositions according to the present invention, these secondary active substances are also significantly more stable in solution. The dispersion compositions described above may be combined with secondary hydrophilic active substances selected from the group comprising native hyaluronan, chondroitin sulfate, glucosamine sulfate, metformin, heparin, inositol, B-group vitamins, lactic acid, amino acids, a-hydroxy and P-hydroxy acids, peptides, water-soluble proteins, carboxymethyl glucan, schizophyllan, glucomannan, panthenol, urea, glycerol, pentylene glycol, and preferably native hyaluronan. Secondary hydrophilic active substances and excipients may be added to the aqueous phase during the preparation of the dispersion composition according to the present invention, or they may be added after the composition has been prepared. Excipients may be selected from the group comprising viscosity regulators, preferably xanthan gum, guar gum, carboxymethylcellulose, sodium alginate, pectin, carbomer, polyethylene glycol, polyvinylpyrrolidone, hydrolyzed collagen, gelatin, and preferably xanthan gum; preservatives selected from a group comprising aromatic acids and their derivatives, preferably benzoic acid, salicylic acid, dehydroacetic acid, potassium sorbate, parabens; alcohols, preferably ethanol, isopropanol, benzyl alcohol, phenoxyethanol, phenethyl alcohol; sugars, preferably sucrose, glucose, fructose, mannose, galactose; salts, preferably sodium chloride.

[0092] The above described dispersion composition according to the present invention may be used directly, or it may be diluted or combined with other substances. For example, the dispersion composition according to the present invention remains stable for at least 1 week at 40 °C after ten-fold dilution with demineralized H2O or an aqueous solution of Zn(Ac)2(Example 1). The above described dispersion composition according to the present invention may be lyophilized and subsequently reconstituted while retaining more than 80% of the active substance originally present in the dispersion composition (Examples 1 and 3; procedure in Example 100). The above described dispersion composition according to the present invention may be combined with other solutions and substances and subsequently converted into a dry form, for example for the preparation of dressings for external or internal wounds. The above described dispersion composition according to the present invention may be applied using various technologies onto dry forms of different materials, such as wound dressings. The abovedescribed dispersion composition according to the present invention may be combined with hydrogels suitable for medical use, for example hydrogels prepared from hyaluronan derivatives. The above described dispersion composition according to the present invention may be incorporated into dry materials comprising nanofibers, microfibers, lyophilizates, nano / microfiber mats, patches, or films suitable for cosmetic or medical applications, for example those based on hyaluronan and other auxiliary polymers.

[0093] The above described dispersion composition according to the present invention may be incorporated into cosmetic and medical creams, gels, emulsions, or serums, which may also contain at least one cosmetic or pharmaceutical excipient selected from the group comprising oil, wax, butter, emulsifier, auxiliary active ingredient, thickener, and preservative. The oil may be selected from the group comprising coconut oil, olive oil, avocado oil, sesame oil, almond oil, castor oil, sunflower oil, hemp oil, jojoba oil, argan oil, apricot oil, borage oil, marula oil, cottonseed oil, evening primrose oil, grape seed oil, hazelnut oil, linseed oil, meadowfoam oil, moringa oil, plum oil, poppy seed oil, rice oil, rosehip oil, safflower oil, wheat germ oil, macadamia oil, and squalene; the butter may be selected from the group comprising cocoa butter, illipe butter, kokum butter, murumuru butter, mango butter, cupuagu butter, avocado butter, and shea butter; the waxes may be selected from the group comprising lanolin, beeswax, carnauba wax, candelilla wax, and petrolatum. The emulsifier may be selected from the group comprising glyceryl caprylate, behenyl alcohol, glyceryl behenate, cetearyl glucoside, methylglucose sesqui stearate, glyceryl stearate citrate, polyglyceryl-3 stearate, cetearyl olivate, lecithin, stearyl alcohol, sorbitan oleate, polysorbates, stearic acid, cetyl alcohol, cetearyl alcohol, sodium acrylate, sodium acryloyldimethyltaurate copolymer, or mixtures thereof.

[0094] The above described dispersion composition according to the present invention may be used for the preparation of oral medical, nutraceutical, and ophthalmological preparations. The dispersion composition according to the present invention may be incorporated into oral medical and nutraceutical products in a liquid form, selected from the group comprising solutions, suspensions, syrups, drops, or emulsions; a gel form, selected from the group comprising hydrogels or jellies; or in a solid form, selected from the group comprising capsules, tablets, or lozenges. Furthermore, it may be used for treatment of the skin or mucous membranes, for cosmetic skin care, for topical application of active substances, for oral delivery of active substances, for dietary supplement applications, or for ophthalmological applications. It may be used in the medical treatment of skin diseases selected from the group comprising atopic dermatitis, psoriasis, ichthyosis, and rosacea, or in the medical treatment of mucosal diseases selected from the group comprising aphthous stomatitis, mucositis, periodontitis,gingivitis, allergic rhinitis, chronic rhinosinusitis, candidiasis, bacterial vaginosis, and Behget’s disease.

[0095] The above described dispersion composition according to the present invention is capable of delivering orders of magnitude higher amounts of curcumin into the skin (0.84 vs.

[0096] 0.05 μg / cm2) (procedure in Example 91) compared to a conventionally prepared dispersion composition (Example 1 vs. Example 50). Using confocal fluorescence microscopy, the differences between the dispersion compositions prepared according to Examples 1, 49, and 50 were confirmed (Fig. 1, top), with curcumin in the sample prepared according to Example 1 showing the best penetration. It was further demonstrated that curcumin in the dispersion composition according to the present invention penetrates in increased amounts into deeper layers of the skin compared to curcumin in oils (Fig. 1). The above described dispersion compositions according to the present invention are also capable of delivering additional secondary active substances into the skin together with curcumin (Examples 36, 37, 39). The ability to deliver active substances into the skin is crucial for cosmetic and dermatological applications.

[0097] In addition to the skin, the dispersion compositions according to the present invention also demonstrate the ability to deliver high amounts of curcumin to the surface of the corneal epithelium (Figs. 2 and 3) and to the oral (Fig. 4 left), nasal (Fig. 4 right), and intestinal (Fig.

[0098] 5) mucosa. For the oral and nasal mucosa, it was shown that curcumin does not remain only on the tissue surface, but penetrates deeper, and in an aqueous solution it is not removed even after one hour of rinsing (Fig. 6). The composition also delivers curcumin into intestinal tissue even after exposure to low pH (Fig. 17). Efficient delivery of active substances to mucosal tissues and the eye is essential for medical, nutraceutical, and ophthalmological applications.

[0099] The dispersion compositions according to the present invention demonstrated antioxidant activity (Examples 1, 36, 37, 39), anti-inflammatory activity (Examples 1 and 39), antimicrobial activity (Example 10), alleviation of ultraviolet (UV)-induced damage (Example 39), wound-healing capability (Examples 1 and 7) thanks to their unique structure, which leads to an enhanced ability to bind curcumin and other active substances necessary for their effective function. The combination with Zn preferably enhances the antioxidant, anti-inflammatory, and antimicrobial properties of curcumin (Prasad 2022).

[0100] For the dispersion compositions according to the present invention prepared according to Examples 1, 36, 37, and 39, antioxidant activity was demonstrated using multiple procedures. In the cell-free DPPH assay (Fig. 7; procedure in Example 93), samples containing curcumin, or curcumin together with additional active substances such as DXM, RST, and CBD, exhibitedsignificantly higher antioxidant activity compared to control samples without curcumin (CTRL, Zn(Ac)2, HA-C18:1 / Zn(Ac)2), and compared to the individual active substances alone (CURC, DXM, RST, CBD). Moreover, samples containing curcumin together with another active substance (HA-C18:1 / Zn(Ac)2+ CURC + DXM (RST)) showed even higher antioxidant activity than the sample containing curcumin alone (HA-C18:1 / Zn(Ac)2+ CURC). Antioxidant activity was also demonstrated in vitro in 3T3 cells (procedure in Example 94), where samples containing curcumin or curcumin with additional active substances reduced cellular oxidative stress to nearly the level of the untreated control (Fig. 8). Control samples without curcumin (HA-C18:1 / H2O, HA-C18:1 / Zn(Ac)2, Zn(Ac)2) only slightly reduced oxidative stress. Samples containing curcumin significantly lowered ROS (reactive oxygen species) levels to those of the untreated control.

[0101] For the dispersion composition according to the present invention prepared according to Example 1, a significant increase in HMOX1 (heme oxygenase-1) gene expression was demonstrated in keratinocytes (procedure in Example 95) compared to the untreated control, 2-MRC (2-mercaptobenzothiazole), and HA-C18:1 / Zn(Ac)2, which indicates an important role of the high curcumin content in the carrier (Fig. 9). Upregulation of HMOX1 is induced in cells under stress conditions (pro-inflammatory or oxidative) as part of the cellular antioxidant defense response (associated, for example, with activation of the Nrf2 pathway) and is therefore generally considered a desirable effect (Shahcheraghi S. H., 2022).

[0102] A potential risk of strong Nrf2 activation leading to HM0X1 expression is the simultaneous high expression of pro-inflammatory cytokines (e.g., IL-1β). This was tested for the dispersion composition according to the present invention prepared according to Example 39, and it was confirmed that the presence of a high amount of curcumin does not increase IL-1β gene expression, thereby preserving the anti-inflammatory activity of dexamethasone (Fig. 10), which is comparable to the effects of free dexamethasone.

[0103] For the dispersion composition according to the present invention (HA-C18:l / Zn(Ac)2 + CURC + DXM) prepared according to Example 39, a significant reduction in IL-8 (interleukin-8) and COX-2 (cyclooxygenase-2) gene expression was demonstrated in ex vivo porcine epidermis (procedure in Example 96) compared to the untreated control (CTRL), HA-C18:l / Zn(Ac)2 alone, and DXM alone (Figs. 11 and 12), which indicates strong anti-inflammatory effects.

[0104] For the dispersion composition according to the present invention containing DXM (HA-C 18: l / Zn(Ac)2 + CURC + DXM) prepared according to Example 39, reduced UV-induced damage was demonstrated in ex vivo porcine epidermis (Figs. 13 and 14; procedure in Example96). In the model of UV-damaged epidermis, UV exposure leads to increased gene expression of matrix metalloproteinase- 1 (MMP-1), responsible for proteolytic degradation of the extracellular matrix, and to decreased expression of superoxide dismutase 2 (SOD2), an important endogenous antioxidant. The carrier containing curcumin and DXM (HA-C18:l / Zn(Ac)2 + CURC + DXM) prepared according to Example 1 reduced MMP-1 expression to the level of the non-irradiated control (CTRL) and increased SOD2 expression compared to the UV-irradiated control (153% of the UV-irradiated control). The combination of curcumin and DXM in the carrier indicates synergistic activity of the individual active substances (CURC, DXM). Thus, the dispersion composition according to the present invention is capable of delivering active substances into the skin, where they exert antioxidant and anti-inflammatory effects.

[0105] The dispersion composition according to the present invention exhibits antimicrobial activity (determination procedure in Example 98) against several microbial species, with microbial counts decreasing over time, reaching zero for most species (Example 10).

[0106] The dispersion compositions according to the present invention prepared according to Examples 1 and 7 demonstrated positive wound-healing activity due to the presence of curcumin or curcumin together with native HA (procedure in Example 97). Results of the scratch assay showed that the treatment solutions promoted migration of HaCaT cells (spontaneously immortalized human keratinocytes) by approximately 15-18 % compared to the negative control after 48 hours (Figs. 15 and 16), for a 400-fold dilution of the solution from Example 1 and a 250-fold dilution of the solution from Example 7.

[0107] In an aqueous environment, the dispersion is composed of particles 10 to 500 nm in size, within which curcumin is encapsulated in high amounts through physical interactions. In addition to the high binding capacity for curcumin, this preparation method also enables the encapsulation of large amounts of certain other active substances, which cannot be encapsulated in such quantities on their own. The dispersions prepared in this manner, with the described composition, exhibit a unique ability to remain stable in solution for extended periods, up to several years, while maintaining a high binding capacity of the encapsulated active substances within the typical storage temperature range (5 to 40 °C). These dispersions also retain their high binding capacity for active substances short-term (2 hours) even under low-pH conditions (pH 1.13). The dispersion compositions according to the present invention are capable of efficiently delivering active substances to selected tissues, such as skin, cornea, or mucosal surfaces, thereby increasing bioavailability and biological efficacy, which can be utilized in pharmaceutical and cosmetic applications.Definitions

[0108] The term “hyaluronan” or “HA” means hyaluronic acid or its pharmaceutically acceptable salt. The term “dispersion composition” means a dispersion system composed of two phases: an aqueous phase and a dispersed phase that is distributed within the solution. The dispersed phase refers to particles preferably containing A-HA, into which curcumin and optionally secondary hydrophobic active substances are encapsulated with the assistance of Zn(Ac)2.

[0109] The term “carrier” means a hydrophobically modified derivative of hyaluronan according to general formula I, preferably acylated hyaluronan (A-HA).

[0110] The term “curcumin” means curcumin according to general formula II, i.e.

[0111] [(1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione],

[0112] demethoxycurcumin, bisdemethoxycurcumin, or a mixture thereof.

[0113] The term “room temperature” means a temperature in the range of 19 °C to 25 °C.

[0114] The term “native hyaluronan” means unmodified hyaluronan or hyaluronic acid.

[0115] Detailed figure descriptions

[0116] Fig. 1: Histological sections of skin (performed according to the procedure described in Example 91) after treatment with the following samples: top left: sample from Example 1, top center: sample from Example 50, top right: sample from Example 49, bottom left: curcumin in almond oil, bottom center: curcumin in jojoba oil, bottom right: negative control (skin autofluorescence).

[0117] Fig. 2: Histological sections of the corneal epithelium with mean fluorescence intensity values (photons / pixel, performed according to the procedure described in Example 92): left: tissue treated with the sample prepared according to Example 1, exposure time 3 minutes; right: negative control (corneal autofluorescence).

[0118] Fig. 3: Scan of the eye surface. Top: surface image with mean fluorescence intensity (photons / pixel, performed according to the procedure described in Example 92), bottom: optical cross-section into the tissue. Left: negative control (eye surface autofluorescence), right: eye treated with the sample prepared according to Example 1, exposure time 1 minute.

[0119] Fig. 4: Histological sections of oral mucosa (left) and nasal mucosa (right) after treatment with a 10-fold diluted sample prepared according to Example 1 (performed according to the procedure described in Example 92), compared with the negative control (oral and nasal mucosaautofluorescence). Exposure time: 1 hour. Values represent mean fluorescence intensity (photons / pixel).

[0120] Fig. 5: Scan of the intestinal mucosal surface. Top: surface image with mean fluorescence intensity (photons / pixel, performed according to the procedure described in Example 92), bottom: optical cross-section into the tissue with mean fluorescence intensity (photons / pixel). Right: negative control (intestinal mucosa autofluorescence), left: intestinal mucosa treated with the sample prepared according to Example 1, exposure time 3 hours.

[0121] Fig. 6: Incubation of nasal (top) and oral (bottom) mucosa with a 10-fold diluted sample prepared according to Example 1, followed by rinsing (performed according to the procedure described in Example 82). Values represent the mean fluorescence intensity of the samples (photons / pixel).

[0122] Fig. 7: DPPH assay (performed according to the procedure described in Example 93). Comparison of the effects of samples containing HA-C18:l / Zn(Ac)2 with CURC, RST, CBD, and DXM prepared according to Examples 1, 36, 37, and 39 on antioxidant activity measured using the DPPH test. Displayed is the % of control (CTRL) from four independent replicates ± standard error of the mean. Student’s t-test: * p<0.05, ** p<0.01, *** p<0.001. No statistically significant differences were observed between the control and Zn(Ac)2, HA-C18:l / Zn(Ac)2, CBD, RST, DXM, or CURC according to the stated criteria.

[0123] Fig. 8: Effect of selected samples (Examples 1, 36, 37, and 39) on oxidative stress in 3T3 cells exposed to H2O2(performed according to the procedure described in Example 94). Displayed is the mean value from two experiments performed in quadruplicate ± standard error of the mean. Tukey’s test: * p<0.05, ** p<0.01. No statistically significant differences were observed between the control, Zn(Ac)2, and HA-C18:1 / H2O according to the stated criteria.

[0124] Fig. 9: HMOX1 gene expression in keratinocytes activated with 2-MRC to induce a pro-inflammatory phenotype (performed according to the procedure described in Example 95). Except for the untreated control, all samples were treated with 2-MRC together with the substances listed in the graph labels. Sensitizing agent 2-MRC alone induces approximately a two-fold increase in HM0X1 expression in keratinocytes, whereas the dispersion composition with curcumin (Example 1) induces an approximately 25-fold increase as part of the cellular protective response to 2-MRC. Displayed is the mean value from four replicates ± standard deviation. Tukey’s test: * p<0.05, ** p<0.01. No statistically significant differences were observed among the remaining combinations according to the stated criteria.

[0125] Fig. 10: IL-1β gene expression in keratinocytes activated with 2-MRC to induce a pro-inflammatory phenotype (performed according to the procedure described in Example 95).Except for the untreated control, all samples were treated with 2-MRC together with the substances listed in the graph labels (Example 38). Displayed is the mean value from four replicates ± standard deviation. Tukey’s test: * p<0.05, ** p<0.01, *** p<0.001. No statistically significant differences were observed among the remaining combinations according to the stated criteria.

[0126] Fig. 11: Effect of curcumin and DXM in the dispersion composition (sample from Example 39) on IL-8 gene expression in ex vivo porcine epidermis 24 hours after treatment (determination according to Example 96). Displayed is the % of control (CTRL) from at least three independent replicates ± standard error of the mean. Tukey’s test: ** p<0.01. No statistically significant differences were observed between the control and HA-C18:l / Zn(Ac)2 or DXM according to the stated criteria.

[0127] Fig. 12: Effect of curcumin and DXM in the dispersion composition (sample from Example 39) on COX-2 gene expression in ex vivo porcine epidermis 24 hours after treatment (determination according to Example 96). Displayed is the % of control (CTRL) from at least three independent replicates ± standard error of the mean. Tukey’s test: * p<0.05, ** p<0.01. No statistically significant differences were observed between the control and HA-C18:l / Zn(Ac)2 or DXM according to the stated criteria.

[0128] Fig. 13: Effect of curcumin and DXM in the dispersion composition (sample from Example 39) on MMP-1 gene expression in UV-irradiated ex vivo porcine epidermis 24 hours after treatment (determination according to Example 96). Displayed is the % of UV-irradiated control (UV CTRL) from at least three independent replicates ± standard error of the mean. Student’s t-test: * p<0.05. No statistically significant differences were observed between the UV-irradiated control andHA-C18:l / Zn(Ac)2, CURC, or DXM according to the stated criteria. Fig. 14: Effect of curcumin and DXM in the dispersion composition (sample from Example 39) on SOD2 gene expression in UV-irradiated ex vivo porcine epidermis 24 hours after treatment (determination according to Example 96). Displayed is the % of UV-irradiated control (UV CTRL) from at least four independent replicates ± standard error of the mean. Student’s t-test: * p<0.05, *** p<0.001. No statistically significant differences were observed between the UV-irradiated control andHA-C18:l / Zn(Ac)2, CURC, or DXM according to the stated criteria. Fig. 15: Scratch assay (performed according to the procedure described in Example 97). Relative density of HaCaT cells in the wound area (%) after 48 hours of migration. HaCaT cells were treated with solutions prepared according to Example 1 (400-fold dilution) and Example 7 (250-fold dilution). As the untreated control (Ctrl-), 0% FBS culture medium was used; as the positive control (Ctrl+), 10% FBS culture medium was used. Displayed are mean valuesfrom quadruplicate measurements within a single experiment ± standard deviation. Analysis performed using IncuCyte 2019B software. Ctrl- is the untreated control. Ctrl+ is the positive control described above.

[0129] Fig. 16: Representative images from the scratch assay image analysis (performed according to the procedure described in Example 97). Shown is wound closure after 48 hours of cell migration (Example 1 at 400-fold dilution and Example 7 at 250-fold dilution). The original wound area is shown in purple; the remaining open wound area is shown in light blue. Analysis performed using IncuCyte 2019B software. Ctrl- is the untreated control; Ctrl+ is the positive control described above.

[0130] Fig. 17: Histological sections of intestinal tissue with mean fluorescence intensity values (photons / pixel, performed according to the procedure described in Example 92) after treatment with the following samples: left: sample from Example 101, center: sample from Example 102, right: negative control (intestinal tissue autofluorescence).

[0131] Examples of embodiments of the invention

[0132] The term HA as used herein refers to hyaluronic acid or its pharmaceutically acceptable salt, preferably Na, K, Ca, Mg, Zn, or Li salts. A-HA refers to acylated HA, such as hexanoyl (C6), lauroyl (C12), palmitoyl (C16), stearoyl (C18), oleoyl (C18:l cis), elaidoyl (C18:l trans), 4-(((2S,3S,4R)-3,4-dihydroxy-2-oleamidooctadecyl)oxy)-4-oxobutanoyl (ceramide), erucoyl (C22:l), Ac acetate, IPA denotes isopropyl alcohol, i.e., propan-2-ol. Unless explicitly stated otherwise, the designation Cl 8:1 refers to the cis conformation (oleoyl).

[0133] HA-Cx:y denotes hyaluronan acylated with various chains. The number following C indicates the number of carbons in the acyl group, and the number after the colon indicates the number of double bonds in the acyl group. If no colon is provided, the acyl group is saturated (contains no double bonds).

[0134] The binding ratio of the active substance relative to the carrier (A-HA), expressed in weight percent and denoted as W, is calculated as Wactive substance Cactive substance in carrier / (Cactive substance in carrier + CCarrier)’ 100, where c denotes concentration in mg / ml or pg / ml. To account for the fact that only the active substance bound to the carrier is considered, the solubility of the active substance in the solvent without A-HA is subtracted as cactive substance in carrier=cactive substance in solution— csolubility of active substance in solvent.The degree of substitution (DS), expressed in %, is defined as the number of acyl groups per 100 HA disaccharides and can be determined, for example, by ¹H NMR spectroscopy (DSNMR, Huerta-Ángeles G., 2014) or gas chromatography (DSGC, Huerta-Ángeles G., 2020).

[0135] Mwis the weight-average molar mass, and for both HA and A-HA it was determined using SEC-MALS (Čožíková D., 2017).

[0136] The solvent-exchange process consists of mixing an aqueous and an organic solvent, followed by evaporation of the more volatile solvent using a rotary evaporator.

[0137] The term phosphate buffer, or PBS, refers to an aqueous solution containing 8 mg / ml NaCl, 0.2 mg / ml KCl, 1.44 mg / ml Na2HPO4, and 0.24 mg / ml KH2PO4, with a pH of 7.4.

[0138] DLS refers to dynamic light scattering. Particle size obtained from DLS is evaluated as the mean value of the particle-size distribution by intensity. The reported DLS particle size corresponds to the mean intensity-weighted particle size and represents at least 95% of the measured signal. If multiple values are reported, this indicates that the particle-size distribution was clearly multimodal and that multiple particle-size populations were present, each contributing more than 5% of the measured signal; their mean sizes and intensity-based percentages are provided.

[0139] Example 1 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0140] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of 55 mM Zn(Ac)2was added to 5 mg of HA-C18:1 with DSGC= 6.8% and Mw= 17.6 x 103g / mol. The solution was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to restore the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0141] Mean particle size from DLS: 240 nm

[0142] Curcumin binding capacity relative to carrier mass: 34.0 wt.%

[0143] Stability of encapsulated curcumin in the dispersion composition: 91.3 % of the initial wt.% relative to carrier mass after 18 months at room temperature and 87.9 % after 3 months at 40 °C and 92 % after 18 months at 5 °CStability after dilution: 93.4 % of the initial wt.% relative to carrier mass after 1 week at 40 °C following 10-fold dilution with demineralized H2O, and 90.6 % after 10-fold dilution with 55 mM Zn(Ac)2.

[0144] Amount of encapsulated curcumin after lyophilization and reconstitution: 84.3 % of the initial wt.% relative to carrier mass.

[0145] Curcumin penetration into skin: 0.84 μg / cm2

[0146] Curcumin penetration into the cornea: 37.4 photon / pixel (negative control 9.5 photon / pixel). Curcumin present on the eye surface: 80.89 photon / pixel (negative control 2.82 photon / pixel). Curcumin penetration into oral mucosa: 26.3 photon / pixel (negative control 0.3 photon / pixel). Curcumin penetration into nasal mucosa: 48.2 photon / pixel (negative control 7.2 photon / pixel). Curcumin penetration into intestinal mucosa: 84.80 photon / pixel (negative control 0.27 photon / pixel).

[0147] Curcumin present on the intestinal surface: 125.9 photon / pixel (negative control 0.6 photon / pixel).

[0148] Antioxidant activity from DPPH assay: 29.7 % of control after 10-fold dilution.

[0149] Antioxidant activity in 3T3 cells: 48 % of the treated control — a value very close to the untreated control (43 %).

[0150] HM0X1 gene expression: 2372 % of untreated control (control treated with 2-MRC: 229 % of untreated control).

[0151] Scratch test: Improvement of HaCaT cell migration (wound healing) by 18 % compared to negative control.

[0152] Example 2 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2with 5 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0153] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of 55 mM Zn(Ac)2was added to 5 mg of HA-C18:1 with DSGC= 6.8% and Mw= 17.6 x 103g / mol. The solution was stirred overnight on a magnetic stirrer at room temperature.

[0154] 10 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to restore the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.Mean particle size from DLS: 207 nm

[0155] Curcumin binding capacity relative to carrier mass: 28.5 wt.%

[0156] Stability of encapsulated curcumin in the dispersion composition: 94.3 % of the initial wt.% relative to carrier mass after 26 months at room temperature.

[0157] Example 3 2 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0158] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at laboratory temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of 55 mM Zn(Ac)2was added to 10 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The solution was stirred overnight on a magnetic stirrer at laboratory temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at 40 °C. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to restore the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0159] Mean particle size from DLS: 240 nm

[0160] Curcumin binding capacity relative to carrier mass: 21.7 wt.%

[0161] Stability of encapsulated curcumin in the dispersion composition: 92.0 % of the initial wt.% relative to carrier mass after 18 months at 5 °C

[0162] Amount of encapsulated curcumin after lyophilization and reconstitution: 91.7 % of the initial wt.% relative to carrier mass

[0163] Example 4 0.3 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process, followed by addition of 0.3 mg / ml 1 x 105g / mol HA Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove any impurities. 5 ml of 55 mM Zn(Ac)2was added to 3 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The solution was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was then replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was filtered through a 1 μm glass filter to remove non-encapsulated curcumin. 5 ml of 55 mM Zn(Ac)2was added to 3 mg of HA with Mw= 100,000 g / mol. The solution was stirred overnight on a magnetic stirrerat laboratory room temperature. This solution was mixed with 5 ml of the HA-C18:1 solution containing encapsulated curcumin.

[0164] Average particle size from DLS: 285 nm

[0165] Curcumin binding capacity relative to carrier mass: 40.9 wt.%

[0166] Stability of encapsulated curcumin in the dispersion: 85.7 % of the initial wt.% relative to carrier mass after 3 months at 40 °C.

[0167] Example 5 1 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process, followed by addition of 0.3 mg / ml 1 x 105g / mol HA Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove any impurities. 5 ml of 55 mM Zn(Ac)2was added to 10 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The solution was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was then replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was filtered through a 1 μm glass filter to remove non-encapsulated curcumin. 5 ml of 55 mM Zn(Ac)2was added to 3 mg of HA with Mw= 100,000 g / mol. The solution was stirred overnight on a magnetic stirrer at room temperature. This solution was mixed with 5 ml of the HA-C18:1 solution containing encapsulated curcumin.

[0168] Average particle size from DLS: 227 nm

[0169] Curcumin binding capacity relative to carrier mass: 20.1 wt.%

[0170] Stability of encapsulated curcumin in the dispersion: 92.2 % of the initial wt.% relative to carrier mass after 18 months at room temperature and 81.0 % after 3 months at 40 °C and 94.0% after 18 months at 5 °C

[0171] Example 6 2 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process, followed by addition of 0.3 mg / ml 1 x 105g / mol HA Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 20 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature usingultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was then replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was filtered through a 1 μm glass filter to remove non-encapsulated curcumin. 5 ml of the 55 mM Zn(Ac)2solution was added to 3 mg of HA with Mw= 100,000 g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. This solution was then mixed with 5 ml of the HA-C18:1 solution containing encapsulated curcumin.

[0172] Average particle size from DLS: 222 nm

[0173] Curcumin binding capacity relative to carrier mass: 11.0 wt.%

[0174] Stability of encapsulated curcumin in the dispersion: 91.8 % of the initial wt.% relative to carrier mass after 3 months at 40 °C

[0175] Example 7 1 mg / ml HA-C18:1 with 0.3 mg / ml 1 x 105g / mol HA in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0176] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 2.5 ml of the 55 mM Zn(Ac)2solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. Another 2.5 ml of the 55 mM Zn(Ac)2solution was added to 1.5 mg of HA with Mw= 1 x 105g / mol. Both solutions were stirred overnight on a magnetic stirrer at room temperature and subsequently combined. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were mixed in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0177] Average particle size from DLS: 259 nm

[0178] Curcumin binding capacity relative to carrier mass: 33.1 wt.%

[0179] Stability of encapsulated curcumin in the dispersion: 98.0 % of the initial wt.% relative to carrier mass after 18 months at room temperature and 89.1 % after 3 months at 40 °C and 98.8% after 18 months at 5 °C.

[0180] Scratch test: Improvement of HaCaT migration (wound healing) by 15 % compared to the negative control.Example 8 2 mg / ml HA-C18:1 with 0.3 mg / ml 1 x 105g / mol HA in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0181] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 2.5 ml of the 55 mM Zn(Ac)2solution was added to 10 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. Another 2.5 ml of the 55 mM Zn(Ac)2solution was added to 1.5 mg of HA-C18:1. Both solutions were stirred overnight on a magnetic stirrer at room temperature and subsequently combined. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were mixed in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. Average particle size from DLS: 255 nm

[0182] Curcumin binding capacity relative to carrier mass: 20.4 wt.%

[0183] Stability of encapsulated curcumin in the dispersion: 93.8 % of the initial wt.% relative to carrier mass after 18 months at room temperature and 89.9 % after 3 months at 40 °C and 94.8% after 18 months at 5 °C.

[0184] Example 9 1 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0185] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 5 mg of HA-C18:1 with DSGC= 6.6 % and Mw= 15.5 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0186] Average particle size from DLS: 280 nm

[0187] Curcumin binding capacity relative to carrier mass: 31.0 wt.%

[0188] Stability of encapsulated curcumin in the dispersion: 93.4 % of the initial wt.% relative to carrier mass after 18 months at room temperature and 83.2 % after 3 months at 40 °C and 93.9% after 18 months at 5 °C.Example 10 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0189] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 100 ml of the 55 mM Zn(Ac)2solution was added to 100 mg of HA-C18:1 with DSGC= 7.1 % and Mw= 17 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 80 mg of curcumin was dissolved in 40 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0190] Average particle size from DLS: 235 nm

[0191] Curcumin binding capacity relative to carrier mass: 33.6 wt.%

[0192] Stability of encapsulated curcumin in the dispersion: 92.8 % of the initial wt.% relative to carrier mass after 18 months at room temperature

[0193] Efficacy against Staphylococcus aureus: t0= 8.3 x 105CFU / ml, after 2 days 0 CFU / ml, after 28 days 0 CFU / ml

[0194] Efficacy against Pseudomonas aeruginosa: t0= 5.4 x 106CFU / ml, after 2 days 0 CFU / ml, after 28 days 0 CFU / ml

[0195] Efficacy against Escherichia coli: t0= 7.3 x 105CFU / ml, after 2 days 0 CFU / ml, after 28 days 0 CFU / ml

[0196] Efficacy against Candida albicans: t0= 4.8 x 106CFU / ml, after 2 days 500 CFU / ml, after 28 days 0 CFU / ml

[0197] Efficacy against Aspergillus brasiliensis: t0= 6.8 x 106CFU / ml, after 2 days 1200 CFU / ml, after 28 days 840 CFU / ml

[0198] A-HA in Zn(Ac)2 with curcumin inhibits the growth of the tested microorganisms over time.

[0199] Example 11 1 mg / ml HA-C6 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0200] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at laboratory temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 5 mg of HA-C6 with DSGC= 35.3 % and Mw= 15 x 103g / mol. The mixture was stirred overnight on a magneticstirrer at laboratory temperature and subsequently sonicated until completely dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at laboratory temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0201] Average particle size from DLS: 178 nm

[0202] Curcumin binding capacity relative to carrier mass: 30.3 wt.%

[0203] Stability of encapsulated curcumin in the dispersion: 93.7 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C.

[0204] Example 12 3 mg / ml HA-C6 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0205] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 15 mg of HA-C6 with DSGC= 16.7 % and Mw= 15 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated until completely dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0206] Average particle size from DLS: 249 nm

[0207] Curcumin binding capacity relative to carrier mass: 13.7 wt.%

[0208] Stability of encapsulated curcumin in the dispersion: 91.4 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C.

[0209] Example 13 3 mg / ml HA-C6 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0210] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 μm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 15 mg of HA-C6 with DSGC= 57.3 % and Mw= 15 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer atroom temperature and subsequently sonicated until completely dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0211] Average particle size from DLS: 177 nm

[0212] Curcumin binding capacity relative to carrier mass: 13.2 wt.%

[0213] Stability of encapsulated curcumin in the dispersion: 80.4 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C.

[0214] Example 14 1 mg / ml HA-C18:1 (trans) in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0215] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 (trans) with DSNMR= 13 % and Mw= 15 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated until completely dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0216] Average particle size from DLS: 269 nm

[0217] Curcumin binding capacity relative to carrier mass: 18.1 wt.%

[0218] Stability of encapsulated curcumin in the dispersion: 82.0 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C.

[0219] Example 15 1 mg / ml HA-C22: 1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0220] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C22:1 with DSNMR= 6.1 % and Mw= 15 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer atroom temperature and subsequently sonicated until completely dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0221] Average particle size from DLS: 252 nm

[0222] Curcumin binding capacity relative to carrier mass: 30.3 wt.%

[0223] Stability of encapsulated curcumin in the dispersion: 90.7 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C

[0224] Example 16 1 mg / ml HA-C18 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0225] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18 with DSNMR=4.7 % and Mw= 13x103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated until fully dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45x102Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0226] Average particle size from DLS: 410 nm

[0227] Curcumin binding capacity relative to carrier mass: 15.4 wt.%

[0228] Stability of encapsulated curcumin in the dispersion: 80.1 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C.

[0229] Example 17 1 mg / ml HA-C16 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0230] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C16 with DSNMR=6.8 % and Mw= 13x103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperatureand subsequently sonicated until fully dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45xl02Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0231] Average particle size from DLS: 292 nm

[0232] Curcumin binding capacity relative to carrier mass: 18.4 wt.%

[0233] Stability of encapsulated curcumin in the dispersion: 89.3 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C.

[0234] Example 18 1 mg / ml HA-ceramide in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0235] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 15 mg of HA-ceramide with DSNMR=3.2 % and Mw= 15x103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated until fully dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45x102Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0236] Average particle size from DLS: 288 nm

[0237] Curcumin binding capacity relative to carrier mass: 28.9 wt.%

[0238] Stability of encapsulated curcumin in the dispersion: 90.0 % of the initial wt.% relative to carrier mass, after 18 months at 5 °C.

[0239] Example 19 0.5 mg / ml HA-C12 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0240] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 2.5 mg of HA-C12 with DSGC=8 % and Mw= 3x105g / mol. The mixture was stirred for 2 days on a magnetic stirrer at room temperature andsubsequently sonicated until fully dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45x102Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0241] Average particle size from DLS: 245 nm

[0242] Curcumin binding capacity relative to carrier mass: 47.7 wt.%

[0243] Stability of encapsulated curcumin in the dispersion: 93.2 % of the initial wt.% relative to carrier mass, after 18 months at 5 °C.

[0244] Example 20 0.5 mg / ml HA-C12 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0245] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 2.5 mg of HA-C12 with DSGC=8 % and Mw= 4.4x105g / mol. The mixture was stirred for 2 days on a magnetic stirrer at laboratory temperature and subsequently sonicated until fully dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at laboratory temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45x102Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0246] Average particle size from DLS: 304 nm

[0247] Curcumin binding capacity relative to carrier mass: 40.5 wt.%

[0248] Stability of encapsulated curcumin in the dispersion: 92.8 % of the initial wt.% relative to carrier mass, after 18 months at 5 °C.

[0249] Example 21 0.5 mg / ml HA-C12 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0250] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 2.5 mg of HA-C12 with DSGC=7 % and Mw= 5.89x105g / mol. The mixture was stirred for 3 days on a magnetic stirrer at room temperatureand subsequently sonicated until fully dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45xl02Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0251] Average particle size from DLS: 278 nm

[0252] Curcumin binding capacity relative to carrier mass: 37.7 wt.%

[0253] Stability of encapsulated curcumin in the dispersion: 90.5 % of the initial wt.% relative to carrier mass, after 18 months at 5 °C.

[0254] Example 22 0.5 mg / ml HA-C6 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0255] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 2.5 mg of HA-C6 with DSNMR=57 % and Mw= 3.5x105g / mol. The mixture was stirred overnight on a magnetic stirrer at laboratory temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45x102Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0256] Average particle size from DLS: 276 nm

[0257] Curcumin binding capacity relative to carrier mass: 43.9 wt.%

[0258] Stability of encapsulated curcumin in the dispersion: 80.1 % of the initial wt.% relative to carrier mass, after 3 months at 40 °C.

[0259] Example 23 1 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 4 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0260] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC=6.8 % and Mw= 17.6x103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.4 mg of curcumin was dissolved in 1 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45x102Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0261] Average particle size from DLS: 164 nm

[0262] Curcumin binding capacity relative to carrier mass: 34.2 wt.%

[0263] Stability of encapsulated curcumin in the dispersion: 94.1 % of the initial wt.% relative to carrier mass, after 18 months at 5 °C.

[0264] Example 24 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)₂ with 3 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0265] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC=6.8 % and Mw= 17.6 x103g / mol. The mixture was stirred overnight on a magnetic stirrer at laboratory temperature. 4.5 mg of curcumin was dissolved in 1.5 ml of IPA at room temperature using ultrasound. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45x102Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0266] Average particle size from DLS: 231 nm

[0267] Curcumin binding capacity relative to carrier mass: 30.2 wt.%

[0268] Stability of encapsulated curcumin in the dispersion: 97.3 % of the initial wt.% relative to carrier mass, after 18 months at 5 °C.

[0269] Example 25 1 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 4 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0270] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC=6.8 % and Mw= 17.6x103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0271] 2 mg of curcumin was dissolved in 0.5 ml of IPA at room temperature using ultrasound. Thesetwo solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under a pressure of 45xl02Pa. The solution was then replenished with demineralized H₂O to restore the original volume of the aqueous phase. The resulting mixture was filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0272] Average particle size from DLS: 183 nm

[0273] Curcumin binding capacity relative to carrier mass: 19.7 wt.%

[0274] Stability of encapsulated curcumin in the dispersion: 91.4 % of the initial wt.% relative to carrier mass, after 18 months at 5 °C.

[0275] Example 26 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)₂ with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process, followed by addition of 0.5 mg / ml xanthan gum Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0276] 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. This solution was added to 2.5 mg of xanthan gum (Mw= 2 x 106g / mol) and stirred overnight at room temperature.

[0277] Mean particle size from DLS: 210 nm

[0278] Curcumin binding capacity relative to carrier mass: 23.3 wt%

[0279] Stability of encapsulated curcumin in the dispersion: 88.1 % of the initial wt% relative to carrier mass, after 3 months at 40 °C

[0280] Example 27 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in ethanol prepared by the solvent-exchange process

[0281] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0282] 4 mg of curcumin was dissolved in 2 ml of ethanol at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotaryevaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0283] Mean particle size from DLS: 243 nm

[0284] Curcumin binding capacity relative to carrier mass: 32.4 wt%

[0285] Stability of encapsulated curcumin in the dispersion: 83.2 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0286] Example 28 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in acetonitrile prepared by the solvent-exchange process

[0287] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0288] 4 mg of curcumin was dissolved in 2 ml of acetonitrile at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0289] Mean particle size from DLS: 308 nm

[0290] Curcumin binding capacity relative to carrier mass: 21.5 wt%

[0291] Stability of encapsulated curcumin in the dispersion: 86.2 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0292] Example 29 1 mg / ml HA-C18:1 with 15 mg / ml 14 x 103g / mol HA in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0293] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities.

[0294] 2.5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. Another 2.5 ml of the 55 mM Zn(Ac)₂ solution was added to 75 mg of HA with Mw= 14 x 103g / mol. Both mixtures were stirred overnight on a magnetic stirrer at room temperature and subsequently combined. 4 mg of curcumin was dissolved in 2 ml of IPA at laboratory temperature using ultrasound. The two solutions were mixed in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solutionwas replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0295] Mean particle size from DLS: 203 nm

[0296] Curcumin binding capacity relative to carrier mass: 32.6 wt%

[0297] Stability of encapsulated curcumin in the dispersion: 89.2 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0298] Example 30 1 mg / ml HA-C18:1 with 5 mg / ml 1.1 x 105g / mol HA in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0299] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities.

[0300] 2.5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. Another 2.5 ml of the 55 mM Zn(Ac)₂ solution was added to 25 mg of HA with Mw= 1.1 x 105g / mol. Both mixtures were stirred overnight on a magnetic stirrer at room temperature and subsequently combined. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were mixed in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. Mean particle size from DLS: 378 nm

[0301] Curcumin binding relative to carrier mass: 28.9 wt%

[0302] Stability of encapsulated curcumin in the dispersion: 89.8 % of the initial wt% relative to carrier mass after 3 months at 40 °C.

[0303] Example 31 1 mg / ml HA-C18:1 with 1 mg / ml 1.52 x 106g / mol HA in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0304] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities.

[0305] 2.5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. Another 2.5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA with Mw= 1.52 x 106g / mol. Both mixtures were stirred overnight on a magnetic stirrer at room temperature and subsequently combined. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were mixed in a flask, and the organicsolvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. Mean particle size from DLS: 355 nm

[0306] Curcumin binding capacity relative to carrier mass: 22.0 wt%

[0307] Stability of encapsulated curcumin in the dispersion: 95.9 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0308] Example 32 1 mg / ml HA-C18:1 in 25 mM Zn(Ac)₂ with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0309] Zn(Ac)₂ was dissolved at a concentration of 25 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 25 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0310] 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0311] Mean particle size from DLS: 225 nm

[0312] Curcumin binding capacity relative to carrier mass: 29.3 wt%

[0313] Stability of encapsulated curcumin in the dispersion: 98.0 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0314] Example 33 1 mg / ml HA-C18:1 in 35 mM Zn(Ac)₂ with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0315] Zn(Ac)₂ was dissolved at a concentration of 35 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 35 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DSGC= 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0316] 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O tothe original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0317] Mean particle size from DLS: 223 nm

[0318] Curcumin binding capacity relative to carrier mass: 24.8 wt%

[0319] Stability of encapsulated curcumin in the dispersion: 93.6 % of the initial wt% relative to carrier mass, after 18 months at 5 °C.

[0320] Example 34 2.5 mg / ml HA-C18:1 in 87.5 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0321] Zn(Ac)? was dissolved at a concentration of 35 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 35 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18: 1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0322] 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0323] Mean particle size from DLS: 283 nm

[0324] Curcumin binding capacity relative to carrier mass: 23.9 wt%

[0325] Stability of encapsulated curcumin in the dispersion: 91.4 % of the initial wt% relative to carrier mass, after 18 months at 5 °C.

[0326] Example 35 1 mg / ml HA-C 18:1 in 200 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0327] Zn(Ac)₂ was dissolved at a concentration of 200 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 200 mM Zn(Ac)2 solution was added to 5 mg of HA-C 18:1 with DSGC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.Mean particle size from DLS: 141 nm

[0328] Curcumin binding relative to carrier mass: 29.8 wt%

[0329] Stability of encapsulated curcumin in the dispersion: 84.9 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0330] Example 36 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin and 2 mg / ml cannabidiol in IPA prepared by the solvent-exchange process

[0331] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18: 1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0332] 2 mg of curcumin was dissolved in 1 ml of IPA at room temperature using ultrasound. 2 mg of cannabidiol was dissolved in 1 ml of IPA at room temperature. These three solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0333] Mean particle size from DLS: 417 nm

[0334] Curcumin binding capacity relative to carrier mass: 18.4 wt%

[0335] Cannabidiol binding capacity relative to carrier mass: 19.4 wt%

[0336] Stability of encapsulated curcumin in the dispersion: 91.1 % of the initial wt% relative to carrier mass, after 3 months at 40 °C

[0337] Stability of cannabidiol in the dispersion: 94.0 % of the initial wt% relative to carrier mass after 3 months at 40 °C

[0338] Curcumin penetration into skin: 1.08 pg / cm2

[0339] Curcumin penetration into acceptor phase: 0.80 pg / ml

[0340] Cannabidiol penetration into skin: 4.39 pg / cm2

[0341] Antioxidant activity (DPPH assay): 49.3 % of control after 10-fold dilution

[0342] Antioxidant activity in 3T3 cells: 45 % of the affected control — a value very close to the unaffected control (43 %).

[0343] Example 37 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin and 2 mg / ml resveratrol in IPA prepared by the solvent-exchange processZn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18: 1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0344] 2 mg of curcumin was dissolved in 1 ml of IPA at room temperature using ultrasound. 2 mg of resveratrol was dissolved in 1 ml of IPA at room temperature. These three solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0345] Mean particle size from DLS: 136 nm

[0346] Curcumin binding capacity relative to carrier mass: 21.8 wt%

[0347] Resveratrol binding capacity relative to carrier mass: 23.7 wt%

[0348] Stability of encapsulated curcumin in the dispersion: 76.1 % of the initial wt% relative to carrier mass, after 3 months at 40 °C

[0349] Stability of encapsulated resveratrol in the dispersion: 70.2 % of the initial wt% relative to carrier mass, after 3 months at 40 °C

[0350] Curcumin penetration into skin: 0.58 pg / cm2

[0351] Resveratrol penetration into skin: 0.80 pg / cm2

[0352] Antioxidant activity (DPPH assay): 19.4 % of control after 10-fold dilution

[0353] Antioxidant activity in 3T3 cells: 48 % of the affected control — a value very close to the unaffected control (43 %).

[0354] Example 38 0.3 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin and 2 mg / ml resveratrol in IPA prepared by the solvent-exchange process

[0355] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)2 solution was added to 1.5 mg of HA-C18:1 with DSGC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 2 mg of curcumin was dissolved in 1 ml of IPA at room temperature using ultrasound. 2 mg of resveratrol was dissolved in 1 ml of IPA at room temperature. These three solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O tothe original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0356] Mean particle size from DLS: 181 nm

[0357] Curcumin binding capacity relative to carrier mass: 40.5 wt%

[0358] Resveratrol binding capacity relative to carrier mass: 42.5 wt%

[0359] Stability of encapsulated curcumin in the dispersion: 83.2 % of the initial wt% relative to carrier mass, after 3 months at 40 °C

[0360] Stability of encapsulated resveratrol in the dispersion: 82.8 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0361] Example 39 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin and 2 mg / ml dexamethasone in IPA prepared by the solvent-exchange process

[0362] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18: 1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0363] 2 mg of curcumin was dissolved in 1 ml of IPA at room temperature using ultrasound. 2 mg of dexamethasone was dissolved in 1 ml of IPA at room temperature. These three solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0364] Mean particle size from DLS: 191 nm

[0365] Curcumin binding capacity relative to carrier mass: 24.4 wt%

[0366] Dexamethasone binding capacity relative to carrier mass: 25.4 wt%

[0367] Stability of encapsulated curcumin in the dispersion: 98.6 % of the initial wt% relative to carrier mass, after 4 months at room temperature

[0368] Stability of encapsulated dexamethasone in the dispersion: 80.1 % of the initial wt% relative to carrier mass, after 4 months at room temperature

[0369] Curcumin penetration into skin: 0.41 pg / cm2

[0370] Dexamethasone penetration into skin: 0.28 pg / cm2

[0371] Antioxidant activity (DPPH assay): 19.9 % of control after 10-fold dilution

[0372] Antioxidant activity in 3T3 cells: 47 % of the affected control — a value very close to the unaffected control (43 %)IL-1β gene expression: 191 % of the unaffected control (control affected with 2-MRC: 303 % of unaffected control) — does not significantly reduce efficacy compared to free dexamethasone (151 % of unaffected control)

[0373] IL-8 gene expression in porcine epidermis: 45 % of control — better than free dexamethasone (84 % of control)

[0374] COX-2 gene expression in porcine epidermis: 13 % of control — better than free dexamethasone (129 % of control)

[0375] MMP-1 gene expression in porcine epidermis: 41 % of UV-irradiated control — better than free dexamethasone (79 % of UV-irradiated control) and slightly better than free curcumin (46 % of UV-irradiated control), approaching the non-irradiated control (49 % of irradiated control)

[0376] SOD2 gene expression: 153 % of UV-irradiated control — better than free dexamethasone (142 % of UV-irradiated control) and slightly better than free curcumin (118 % of

[0377] UV-irradiated control), among the tested samples the closest to the non-irradiated control (353 % of irradiated control).

[0378] Example 40 0.3 mg / ml HA-C18:1 in 55 mM Zn(Ac)₂ with 2 mg / ml curcumin and 2 mg / ml dexamethasone in IPA prepared by the solvent-exchange process

[0379] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 1.5 mg of HA-C18:1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 2 mg of curcumin was dissolved in 1 ml of IPA at room temperature using ultrasound. 2 mg of dexamethasone was dissolved in 1 ml of IPA at room temperature. These three solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0380] Mean particle size from DLS: 360 nm

[0381] Curcumin binding capacity relative to carrier mass: 45.8 wt%

[0382] Dexamethasone binding capacity relative to carrier mass: 54.3 wt%

[0383] Stability of encapsulated curcumin in the dispersion: 99.2 % of the initial wt% relative to carrier mass, after 4 months at room temperatureStability of encapsulated dexamethasone in the dispersion: 92.4 % of the initial wt% relative to carrier mass, after 4 months at room temperature

[0384] Example 41 0.6 mg / ml HA-C18:1 with 1.2 mg / ml 1.17 x 106g / mol HA in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0385] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities.

[0386] 2.5 ml of the 55 mM Zn(Ac)₂ solution was added to 3 mg of HA-C18: 1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. Another 2.5 ml of the 55 mM Zn(Ac)2 solution was added to 6 mg of HA with Mw= 1.17 x 106g / mol. Both mixtures were stirred overnight on a magnetic stirrer at room temperature and subsequently combined. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin, followed by filtration through a 0.22 pm nylon filter to sterilize the dispersion. Mean particle size from DLS before 0.22 pm filtration: 455 nm

[0387] Curcumin binding capacity relative to carrier mass before 0.22 pm filtration: 32 wt% Curcumin binding capacity relative to carrier mass after 0.22 pm filtration: 22.9 wt% Stability of encapsulated curcumin in the dispersion: 82.4 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0388] Example 42 1 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process and filtered through a 0.22 pm filter

[0389] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18: 1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 0.22 pm nylon filter to remove non-encapsulated curcumin and to sterilize the solution.Mean particle size from DLS: 219 nm

[0390] Curcumin binding capacity relative to carrier mass: 24.7 wt%

[0391] Stability of encapsulated curcumin in the dispersion: 91.8 % of the initial wt% relative to carrier mass, after 18 months at room temperature.

[0392] Example 43 3 mg / ml HA-C18: 1 in 55 mM Zn(Ac)₂ with 4 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0393] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 15 mg of HA-C18:1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 8 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0394] Mean particle size from DLS: 264 nm

[0395] Curcumin binding capacity relative to carrier mass: 23 wt%

[0396] Stability of encapsulated curcumin in the dispersion: 93.6 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0397] Example 44 4 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 4 mg / ml curcumin in IPA prepared by the solvent-exchange process

[0398] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 20 mg of HA-C18:1 with DS_GC = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 8 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0399] Mean particle size from DLS: 263 nm

[0400] Curcumin binding capacity relative to carrier mass: 20.2 wt%Stability of encapsulated curcumin in the dispersion: 95.3 % of the initial wt% relative to carrier mass, after 3 months at 40 °C.

[0401] Example 45 1 mg / ml HA-C18: 1 in 20 mM Zn(Ac)₂ with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0402] Zn(Ac)₂ was dissolved at a concentration of 20 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 20 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0403] 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0404] Mean particle size from DLS: 393 nm

[0405] Curcumin binding capacity relative to carrier mass: 19.4 wt%

[0406] Stability of encapsulated curcumin in the dispersion: 77.1 % of the initial wt% relative to carrier mass, after 3 weeks at 40 °C.

[0407] Curcumin was not stable in the solution.

[0408] Example 46 0.5 mg / ml HA-C12 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0409] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 2.5 mg of HA-C12 with DS = 8 % and Mw= 7.94 x 105g / mol. The mixture was stirred for 7 days on a magnetic stirrer at room temperature and subsequently sonicated until fully dissolved. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0410] Mean particle size from DLS: 350 nm

[0411] Curcumin binding capacity relative to carrier mass: 26.3 wt%Stability of encapsulated curcumin in the dispersion: 81.9 % of the initial wt% relative to carrier mass, after 16 months at 5 °C.

[0412] Curcumin was not stable in the solution.

[0413] Example 47 1 mg / ml HA-C18:1 in 55 mM zinc gluconate with 2 mg / ml curcumin in IP A prepared by the solvent-exchange process (comparative example)

[0414] Zinc gluconate was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 55 mM zinc gluconate solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at laboratory temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0415] Mean particle size from DLS: 339 nm

[0416] Curcumin binding capacity relative to carrier mass: 20.5 wt%

[0417] Stability of encapsulated curcumin in the dispersion: 86.7 % of the initial wt% relative to carrier mass, after 2 months at 5 °C.

[0418] Curcumin was not stable in the solution.

[0419] Example 48 1 mg / ml HA-C18:1 in 1.6 mM zinc citrate with 2 mg / ml curcumin in IP A prepared by the solvent-exchange process (comparative example)

[0420] Zinc citrate was dissolved at a concentration of 1.6 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove any impurities. 5 ml of the 1.6 mM zinc citrate solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0421] Mean particle size from DLS: 457 nm

[0422] Curcumin binding capacity relative to carrier mass: 2.4 wt%Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity.

[0423] Curcumin was not encapsulated in a sufficient amount.

[0424] Example 49 1 mg / ml HA-C18:1 in demineralized water with 0.8 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0425] 5 ml of demineralized water was added to 5 mg of HA-C18: 1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 1.6 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0426] Mean particle size from DLS: 263 nm

[0427] Curcumin binding capacity relative to carrier mass: 0.1 wt%

[0428] Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity

[0429] Curcumin was not encapsulated in a sufficient amount.

[0430] Example 50 1 mg / ml HA-C18:1 in 0.15 M PBS with 0.8 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0431] 5 ml of 0.15 M PBS was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 1.6 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0432] Mean particle size from DLS: two particle-size distributions — 80 % at 270 nm, 20 % at 37 nm

[0433] Curcumin binding capacity relative to carrier mass: 1.6 wt%

[0434] Stability of encapsulated curcumin in the dispersion: 53.4 % of the initial wt% relative to carrier mass, after 4 days at 5 °C

[0435] Skin penetration: 0.05 μg / cm2Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0436] Example 51 1 mg / ml HA-C18:1 in 41.6 mM ZnSO4 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0437] ZnSO4 was dissolved at a concentration of 41.6 mM in demineralized H2O at room temperature.

[0438] 5 ml of the 41.6mM ZnSO4solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at laboratory temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at laboratory temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0439] Mean particle size from DLS: two particle-size distributions — 89 % at 287 nm, 11% at 32 nm

[0440] Curcumin binding capacity relative to carrier mass: 2.2 wt%

[0441] Stability of encapsulated curcumin in the dispersion: 77.7 % of the initial wt% relative to carrier mass, after 3 days at 5 °C.

[0442] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0443] Example 52 1 mg / ml HA-C 18: 1 in 55.5 mM Ca(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0444] Ca(Ac)? was dissolved at a concentration of 55.5 mM in demineralized EEO at room temperature. 5 ml of the 55.5 mM Ca(Ac)2 solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0445] Mean particle size from DLS: two particle-size distributions — 80 % at 295 nm, 18 % at 34 nm

[0446] Curcumin binding capacity relative to carrier mass: 1.1 wt%

[0447] Stability of encapsulated curcumin in the dispersion: 68.8 % of the initial wt% relative to carrier mass, after 3 days at 5 °C.Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0448] Example 53 1 mg / ml HA-C18:1 in 55.5 mM Mg(Ac)2with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0449] Mg(Ac)2was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Mg(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0450] Mean particle size from DLS: 253 nm

[0451] Curcumin binding capacity relative to carrier mass: 1.4 wt%

[0452] Stability of encapsulated curcumin in the dispersion: 66.7 % of the initial wt% relative to carrier mass, after 2 weeks at 5 °C.

[0453] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0454] Example 54 1 mg / ml HA-C18:1 in 166 mM NaAc with 5 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0455] NaAc was dissolved at a concentration of 166 mM in demineralized H2O at room temperature.

[0456] 5 ml of the 166 mM NaAc solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0457] 10 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0458] Mean particle size from DLS: two particle-size distributions — 89 % at 331 nm, 10 % at 33 nm

[0459] Curcumin binding capacity relative to carrier mass: 0.5 wt%

[0460] Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity.

[0461] Curcumin was not encapsulated in a sufficient amount.Example 55 1 mg / ml HA-C18:1 in 166 mM NaCl with 5 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0462] NaCl was dissolved at a concentration of 166 mM in demineralized H2O at room temperature.

[0463] 5 ml of the 166 mM NaCl solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature.

[0464] 10 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0465] Mean particle size from DLS: 253 nm

[0466] Curcumin binding capacity relative to carrier mass: 0.5 wt%

[0467] Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity.

[0468] Curcumin was not encapsulated in a sufficient amount.

[0469] Example 56 1 mg / ml HA-C18:1 in 55.5 mM Cd(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0470] Cd(Ac)2was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Cd(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0471] Mean particle size from DLS: 326 nm

[0472] Curcumin binding capacity relative to carrier mass: 7.9 wt%

[0473] Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity.

[0474] Curcumin was not encapsulated in a sufficient amount.

[0475] Example 57 1 mg / ml HA-C18: 1 in 55.5 mM Co(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)Co(Ac)2was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Co(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0476] Mean particle size from DLS: 183 nm

[0477] Curcumin binding capacity relative to carrier mass: 22.8 wt%

[0478] Stability of encapsulated curcumin in the dispersion: 57.8 % of the initial wt% relative to carrier mass, after 6 months at 5 °C.

[0479] Curcumin was not stable in the solution.

[0480] Example 58 1 mg / ml HA-C18: 1 in 55.5 mM ZnC104 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0481] ZnClO4was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM ZnClO4solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0482] Mean particle size from DLS: 158 nm

[0483] Curcumin binding capacity relative to carrier mass: 0.7 wt%

[0484] Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity.

[0485] Curcumin was not encapsulated in a sufficient amount.

[0486] Example 59 1 mg / ml HA-C18:1 in 55.5 mM Mn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0487] Mn(Ac)2was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Mn(Ac)2solution was added to 5 mg of HA-C18:1 with DS= 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0488] Mean particle size from DLS: 425 nm

[0489] Curcumin binding capacity relative to carrier mass: 16.3 wt%

[0490] Stability of encapsulated curcumin in the dispersion: 73.8 % of the initial wt% relative to carrier mass after 7 days at 40 °C.

[0491] Curcumin was not stable in the solution.

[0492] Example 60 1 mg / ml HA-C 18: 1 in 5.55 mM Cu(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0493] Cu(Ac)2was dissolved at a concentration of 5.55 mM in demineralized H2O at room temperature. 5 ml of the 5.55 mM Cu(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0494] Mean particle size from DLS: 793 nm

[0495] Curcumin binding capacity relative to carrier mass: 16.7 wt%

[0496] Stability of encapsulated curcumin in the dispersion: 77.5 % of the initial wt% relative to carrier mass, after 2 months at 40 °C.

[0497] Curcumin was not stable in the solution and formed excessively large particles.

[0498] Example 61 1 mg / ml HA-C 18: 1 in 55.5 mM Yb(Ac)s with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0499] Yb(Ac)3was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Yb(Ac)3solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated until fully dissolved. 4 mg of curcumin wasdissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0500] Mean particle size from DLS: 1548 nm

[0501] Curcumin binding capacity relative to carrier mass: 2.5 wt%

[0502] Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity.

[0503] Curcumin was not encapsulated in a sufficient amount and formed excessively large particles.

[0504] Example 62 1 mg / ml HA-C12 in 55.5 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0505] Zn(Ac)2was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Zn(Ac)2solution was added to 5 mg of HA-C12 with DS = 23.7 % and Mw= 15 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0506] Mean particle size from DLS: 1629 nm

[0507] Curcumin binding capacity relative to carrier mass: 0.2 wt%

[0508] Stability of encapsulated curcumin in the dispersion: not monitored due to low binding capacity.

[0509] Curcumin was not encapsulated in a sufficient amount and formed excessively large particles.

[0510] Example 63 5 mg / ml HA-C 18: 1 in 55.5 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0511] Zn(Ac)2was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Zn(Ac)2solution was added to 25 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated. 4 mg of curcumin was dissolved in 2 ml of IPAat room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0512] Mean particle size from DLS: 286 nm

[0513] Curcumin binding capacity relative to carrier mass: 7.8 wt%

[0514] Stability of encapsulated curcumin in the dispersion: 77.9 % of the initial wt% relative to carrier mass, after 4 months at room temperature.

[0515] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0516] Example 64 10 mg / ml HA-C18:1 in 55.5 mM Zn(Ac)2 with 2 mg / ml curcumin in IP A prepared by the solvent-exchange process (comparative example)

[0517] Zn(Ac)? was dissolved at a concentration of 55.5 mM in demineralized H2O at room temperature. 5 ml of the 55.5 mM Zn(Ac)2 solution was added to 50 mg of HA-C18: 1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated. 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0518] Mean particle size from DLS: 175 nm

[0519] Curcumin binding capacity relative to carrier mass: 5.6 wt%

[0520] Stability of encapsulated curcumin in the dispersion: 50.5 % of the initial wt% relative to carrier mass, after 4 months at laboratory temperature.

[0521] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0522] Example 65 1 mg / ml HA-C18:1 in 11.1 mM Zn(Ac)2with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0523] Zn(Ac)2was dissolved at a concentration of 11.1 mM in demineralized H2O at room temperature. 5 ml of the 11.1 mM Zn(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature and subsequently sonicated. 4 mg of curcumin was dissolved in 2 ml of IPAat room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0524] Mean particle size from DLS: 244 nm

[0525] Curcumin binding capacity relative to carrier mass: 28.8 wt%

[0526] Stability of encapsulated curcumin in the dispersion: 53.7 % of the initial wt% relative to carrier mass after 3 months at laboratory temperature and 79.8 % after 1 week at 40 °C and 73.1 % after 3 months at 5 °C

[0527] Curcumin was not stable in the solution.

[0528] Example 66 1 mg / ml HA-C18: 1 in 55 mM Zn(Ac)2 with 0.7 mg / ml curcumin prepared by the direct-dissolution process (comparative example)

[0529] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 3 ml of the 55 mM Zn(Ac)2solution was added to 3 mg of HA-C18:1 with DS = 6.8% and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. This solution was then transferred into a vial containing 2.1 mg of curcumin. The vial was shaken for 48 hours on a shaker at 150 rpm at 25 °C. The resulting mixture was subsequently filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0530] Mean particle size from DLS: two particle-size distributions — 92 % at 261 nm, 8 % at 37 nm Curcumin binding capacity relative to carrier mass: 1.0 wt%

[0531] Stability of encapsulated curcumin in the dispersion: 73.4 % of the initial wt% relative to carrier mass, after 38 days at 5 °C.

[0532] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0533] Example 67 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 0.7 mg / ml curcumin prepared by the IPA-film process (comparative example)

[0534] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 3 ml of the 55 mM Zn(Ac)2solution was added to 3 mg of HA-C18:1 with DS = 6.8% and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. Separately, 2.1 mg of curcumin was dissolved in 3 ml of IPA in a vial. TheIPA was evaporated under a gentle stream of nitrogen at atmospheric pressure, forming a thin curcumin film in the vial. The HA-C18:1 solution in Zn(Ac)2 was then pipetted onto the curcumin film. The vial was shaken for 48 hours on a shaker at 150 rpm at 25 °C. The resulting mixture was subsequently filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0535] Mean particle size from DLS: two particle-size distributions — 92 % at 277 nm, 8 % at 35 nm Curcumin binding capacity relative to carrier mass: 1.2 wt%

[0536] Stability of encapsulated curcumin in the dispersion: 74.7 % of the initial wt% relative to carrier mass, after 38 days at 5 °C.

[0537] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0538] Example 68 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in acetone prepared by the solvent-exchange process (comparative example)

[0539] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8% and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature using ultrasound. Separately, 4 mg of curcumin was dissolved in 2 ml of acetone at room temperature. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. Mean particle size from DLS: 456 nm

[0540] Curcumin binding capacity relative to carrier mass: 37.5 wt%

[0541] Stability of encapsulated curcumin in the dispersion: 63.8 % of the initial wt% relative to carrier mass, after 1 week at 40 °C.

[0542] Curcumin was not stable in the solution.

[0543] Example 69 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in tetrahydrofuran prepared by the solvent-exchange process (comparative example) Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8% and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer atroom temperature. Separately, 4 mg of curcumin was dissolved in 2 ml of tetrahydrofuran at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. Mean particle size from DLS: 484 nm

[0544] Curcumin binding capacity relative to carrier mass: 1.5 wt%

[0545] Stability of encapsulated curcumin in the dispersion: 57.7 % of the initial wt% relative to carrier mass, after 5 days at 40 °C.

[0546] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0547] Example 70 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in chloroform prepared by the solvent-exchange process (comparative example)

[0548] Zn(Ac)2was dissolved at a concentration of 55 mM in demineralized H2O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2solution was added to 5 mg of HA-C18:1 with DS = 6.8% and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. Separately, 4 mg of curcumin was dissolved in 2 ml of chloroform at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H2O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. Mean particle size from DLS: 381 nm

[0549] Curcumin binding capacity relative to carrier mass: 2.3 wt%

[0550] Stability of encapsulated curcumin in the dispersion: 51.5 % of the initial wt% relative to carrier mass, after 5 days at 40 °C.

[0551] Curcumin was not encapsulated in a sufficient amount and was not stable in the solution.

[0552] Example 71 1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in methanol prepared by the solvent-exchange process (comparative example)

[0553] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)2 solution was added to 5 mg of HA-C18:1 with DS = 6.8% and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer atroom temperature. Separately, 4 mg of curcumin was dissolved in 2 ml of methanol at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin. Mean particle size from DLS: 313 nm

[0554] Curcumin binding capacity relative to carrier mass: 29 wt%

[0555] Stability of encapsulated curcumin in the dispersion: 76.1 % of the initial wt% relative to carrier mass, after 9 weeks at 40 °C.

[0556] Curcumin was not stable in the solution.

[0557] Example 72 10 mg / ml 15 x 103g / mol HA in 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0558] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. 5 ml of the 55 mM Zn(Ac)₂ solution was added to 50 mg of HA with Mw= 15 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. Separately, 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0559] Amount of curcumin in the dispersion: 8 pg / ml

[0560] Stability of encapsulated curcumin: not monitored due to low binding capacity

[0561] Curcumin was not encapsulated in a sufficient amount.

[0562] Example 73 55 mM Zn(Ac)2 with 2 mg / ml curcumin in IPA prepared by the solvent-exchange process (comparative example)

[0563] Zn(Ac)₂ was dissolved at a concentration of 55 mM in demineralized H₂O at room temperature. The solution was filtered through a 0.2 pm nylon membrane filter to remove potential impurities. Separately, 4 mg of curcumin was dissolved in 2 ml of IPA at room temperature using ultrasound. This solution was mixed in a flask with 5 ml of the 55 mM Zn(Ac)₂ solution, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase.The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0564] Amount of curcumin in the dispersion: 3 pg / ml

[0565] Stability of encapsulated curcumin: not monitored due to low binding capacity

[0566] Curcumin was not encapsulated in a sufficient amount.

[0567] Example 74 1 mg / ml HA-C18:1 in 55.5 mM Zn(Ac)2 with 2 mg / ml resveratrol in IPA prepared by the solvent-exchange process (comparative example)

[0568] Zn(Ac)₂ was dissolved at a concentration of 55.5 mM in demineralized H₂O at room temperature. 5 ml of the 55.5 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. Separately, 4 mg of resveratrol was dissolved in 2 ml of IPA at room temperature. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated resveratrol.

[0569] Mean particle size from DLS: two particle-size distributions — 93 % at 240 nm, 7 % at 35 nm Resveratrol binding capacity relative to carrier mass: 0 wt% — encapsulation was equivalent to dissolution in solvent.

[0570] Resveratrol was not encapsulated in a sufficient amount.

[0571] Example 75 1 mg / ml HA-C18:1 in 55.5 mM Zn(Ac)2 with 2 mg / ml cannabidiol in IPA prepared by the solvent-exchange process (comparative example)

[0572] Zn(Ac)₂ was dissolved at a concentration of 55.5 mM in demineralized H₂O at room temperature. 5 ml of the 55.5 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The mixture was stirred overnight on a magnetic stirrer at room temperature. Separately, 4 mg of cannabidiol was dissolved in 2 ml of IPA at room temperature. The two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C and 45 x 102Pa. The solution was replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting dispersion was then filtered through a 1 μm glass filter to remove non-encapsulated cannabidiol.

[0573] Mean particle size from DLS: 513 nm

[0574] Cannabidiol binding capacity relative to carrier mass: 8.1 wt%Stability of encapsulated cannabidiol in the dispersion: 39.6 % of the initial wt% relative to carrier mass, after 6 weeks at 5 °C.

[0575] Cannabidiol was not encapsulated in a sufficient amount, was not stable in the solution, and formed excessively large particles.

[0576] Example 76 1 mg / ml HA-C18:1 in 55.5 mM Zn(Ac)2 with 2 mg / ml dexamethasone in IPA prepared by the solvent-exchange process (comparative example)

[0577] Zn(Ac)₂ was dissolved at a concentration of 55.5 mM in demineralized H₂O at room temperature. 5 ml of the 55.5 mM Zn(Ac)₂ solution was added to 5 mg of HA-C18:1 with DS = 6.8 % and Mw= 17.6 x 103g / mol. The solution was stirred overnight on a magnetic stirrer at room temperature. 4 mg of dexamethasone was dissolved in 2 ml of IPA at room temperature. These two solutions were combined in a flask, and the organic solvent was evaporated on a rotary evaporator at 34 °C under 45 x 102Pa. The solution was then replenished with demineralized H₂O to the original volume of the aqueous phase. The resulting solution was subsequently filtered through a 1 μm glass filter to remove non-encapsulated curcumin.

[0578] Average particle size from DLS:

[0579] Two particle-size distributions - 94 % at 462 nm, 6 % at 45 nm.

[0580] Dexamethasone binding capacity relative to carrier mass: 0 wt% - encapsulation was the same as in the solvent.

[0581] Dexamethasone was not encapsulated in a sufficient amount.

[0582] Example 77 Emulsion containing the dispersion composition and linseed oil

[0583] 50 µl of the dispersion composition prepared according to Example 1 was mixed by vortexing for 1 minute with 1 ml of demineralized H₂O and 50 pl of polysorbate 80. Subsequently, 15 pl of linseed oil was added. The resulting mixture was vortexed for 1 minute to form an emulsion containing the dispersion composition.

[0584] Example 78 Emulsion containing the dispersion composition and jojoba oil

[0585] 50 µl of the dispersion composition prepared according to Example 1 was mixed by vortexing for 1 minute with 1 ml of demineralized H₂O and 50 pl of polysorbate 80. Subsequently, 15 pl of jojoba oil was added. The resulting mixture was vortexed for 1 minute to form an emulsion containing the dispersion composition.

[0586] Example 79 Emulsion containing the dispersion composition and castor oil50 µl of the dispersion composition prepared according to Example 1 was mixed by vortexing for 1 minute with 1 ml of demineralized H₂O and 50 pl of polysorbate 80. Subsequently, 5 pl of castor oil was added. The resulting mixture was vortexed for 1 minute to form an emulsion containing the dispersion composition.

[0587] Example 80 Serum containing the dispersion composition with niacinamide

[0588] 50 mg of niacinamide was dissolved in 1 ml of demineralized H₂O. This solution was mixed by vortexing for 1 minute with 50 µl of the dispersion composition prepared according to Example 1, resulting in a serum containing the dispersion composition.

[0589] Example 81 Serum containing the dispersion composition with low-molecular-weight native hyaluronan

[0590] 5 mg of native HA with a molecular weight of 15 x 106g mol-1was dissolved in 1 ml of demineralized ECO. This solution was mixed by vortexing for 1 minute with 100 pl of the dispersion composition prepared according to Example 1, resulting in a serum containing the dispersion composition.

[0591] Example 82 Serum containing the dispersion composition with high-molecular-weight native hyaluronan

[0592] 8 mg of native HA with a molecular weight of 1.5 x 106g mol-1was dissolved in 1 ml of demineralized H₂O. This solution was mixed with 50 µl of the dispersion composition prepared according to Example 1 by stirring for 30 minutes on a magnetic stirrer at room temperature, resulting in a serum containing the dispersion composition.

[0593] Example 83 Serum containing the dispersion composition with a combination of low- and high-molecular-weight native hyaluronan

[0594] 8 mg of native HA with a molecular weight of 1.5 x 106g mol-1was dissolved in 1 ml of demineralized H₂O. 10 mg of 15 kDa native HA was dissolved in 0.5 ml of demineralized H₂O. These solutions were mixed with 50 µl of the dispersion composition prepared according to Example 1 by stirring for 30 minutes on a magnetic stirrer at room temperature, resulting in a serum.

[0595] Example 84 Cream containing the dispersion compositionInto the first beaker, 1.65 g of olive oil, 0.15 g of sorbitan stearate, 0.25 g of polysorbate 60, 0.4 g of cetyl alcohol, and 0.2 g of stearic acid were weighed. Into the second beaker, 7.25 g of demineralized water and 0.02 g of EDTA were weighed. Both beakers were heated in a water bath to 70 °C until all components were dissolved. Under continuous stirring, the contents of the second beaker were poured into the first beaker. After cooling the mixture to 35 °C, 0.08 g of benzyl alcohol was added to the first beaker under continuous stirring. After cooling down to room temperature, 0.5 g of the dispersion composition prepared according to Example 1 was incorporated into the resulting cream, yielding a cream containing the dispersion composition.

[0596] Example 85 Gel containing the dispersion composition

[0597] 20 mg of xanthan gum was dissolved in 2 ml of demineralized H₂O and mixed with 100 pl of the dispersion composition prepared according to Example 1 by stirring overnight on a magnetic stirrer at room temperature, resulting in a gel containing the dispersion composition.

[0598] Example 86 Determination of particle size by dynamic light scattering (DLS), general procedure

[0599] The hydrodynamic particle size was determined using dynamic light scattering (DLS) at 25 °C on a Zetasizer Nano-ZS instrument (Malvern Instruments) equipped with a 633 nm He-Ne laser and a dynamic light-scattering detector after 24 h equilibration. Particle size in solution was calculated using Zetasizer Software 7.11, and the particle-size distribution was evaluated based on intensity. As a result, the reported value corresponds to the mean particle size by intensity and represents at least 95 % of the measured signal. If multiple values are reported, this indicates that the particle-size distribution was clearly multimodal and that several particle populations were present, each contributing more than 5 % of the measured signal. Their mean sizes and percentage representation by intensity are provided.

[0600] Example 87 Determination of curcumin binding, general procedure

[0601] The amount of dissolved curcumin was determined from a calibration curve measured using an absorption spectrometer at 425 nm and is expressed as the weight percentage of the active substance relative to the mass of the carrier. The absorbance of the curcumin-containing sample was corrected by subtracting the absorbance of a blank sample without curcumin. Additionally, the solubility of curcumin in the solvent (Example 72) was subtracted. Highly bound samples had to be diluted multiple times to enable accurate quantification.Example 88 Determination of dexamethasone binding, general procedure

[0602] For the determination of dexamethasone content in the samples, a liquid chromatograph Acquity H-Class UPLC system (Waters) equipped with a photodiode array (PDA) detector (Waters) was used. Separation was performed on a reverse-phase column Jupiter 4 p Proteo 90 A (150 x 4.6 mm, Phenomenex) at a constant temperature in the column oven (35 °C) and autosampler (10 °C). The binary mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B). The mobile-phase flow rate was set to 0.8 ml / min with the following gradient: B starts at 40 % and is held for 0.5 min, increased to 55 % over 1.5 min, increased to 60 % over 7 min, increased to 85 % over 1 min and held for 1 min, decreased back to 40 % over 0.5 min and held for 3 min The injection volume was 10 pl. The PDA detector was set to a wavelength of 240 nm. Empower software was used for data acquisition and processing.

[0603] Example 89 Determination of resveratrol binding, general procedure

[0604] For the determination of resveratrol content in the samples, a liquid chromatograph Acquity H-Class UPLC system (Waters) equipped with a PDA detector (Waters) was used. Separation was performed on a reverse-phase column Jupiter 4 p Proteo 90 A (150 x 4.6 mm, Phenomenex) at a constant temperature in the column oven (30 °C) and autosampler (10 °C). The binary mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B). The mobile-phase flow rate was set to 0.8 ml / min with the following gradient: B starts at 40 % and is held for 1 min, increased to 55 % over 1 min, increased to 60 % over 7 min, increased to 85 % over 1 min and held for 1 min, decreased to 40 % over 0.5 min and held for 3 min. The injection volume was 5 pl. The PDA detector was set to a wavelength of 306 nm. Empower software was used for data acquisition and processing.

[0605] Example 90 Determination of cannabidiol binding, general procedure

[0606] For the determination of cannabidiol content in the samples, a liquid chromatograph Alliance HPLC system (2695 Separation Module, Waters) equipped with a PDA detector (2998 Photodiode Array, Waters) was used. Separation was performed on a reverse-phase column KINETEX C18 XB 5 pm (250 x 4.6 mm) using mobile phases A (0.1% trifluoroacetic acid in water) and B (acetonitrile), at a constant temperature in the column oven (30 °C) and autosampler (10 °C). The mobile-phase flow rate was set to 1.0 ml / min with the following gradient: B starts at 60 %, increased to 95 % over 8 min, held at 95 % for 1 min, increased to 100 % over 0.5 min and held for 3.5 min, decreased to 60 % over 1 min and held for 5 min. Theinjection volume was 15 pl. The PDA detector was set to a wavelength of 240 nm. Empower software was used for data acquisition and processing.

[0607] Example 91 Determination of penetration of active substances into the skin, general procedure

[0608] Skin-penetration tests were performed in Franz diffusion cells according to EFSA guidelines (European Food Safety et al., 2017). Skin from porcine ear auricles obtained from a slaughterhouse (Bocus, Letohrad, Czech Republic) was processed using a dermatome to obtain 600 pm-thick slices, which were mounted in the diffusion cells with the stratum comeum facing upward. After 30 minutes of equilibration, the diffusion area (0.64 cm2) was filled with 130 pl of the sample solution and covered. BSA (40 mg / ml) was used as the acceptor medium at 32 °C. Penetration experiments were carried out for 24 hours. Afterwards, the skin samples were removed from the cells, rinsed with buffer (PBS, 0.15 M, pH 7.4), and dried. For quantitative analysis, donor and acceptor fluids were collected, tape-stripping was performed on the skin surface, and the remaining skin was extracted with a mixture of organic solvent and water. Samples were analyzed by HPLC-UV to determine the amount of active substance in the skin and acceptor medium. Penetrated amounts of active substances into the skin (and, where applicable, into the acceptor medium) are reported in Examples 1, 35, 36, 38, and 49. For microscopic evaluation, tissue samples were frozen in OCT (optimal cutting temperature) medium, and sectioned into 10 pm slices using a Leica CM1950 cryostat (Leica Biosystems, Germany). Fluorescence images were acquired using a Leica TCS SP8 X confocal microscope (Leica Microsystems) equipped with a Leica HC PL APO CS2 objective (63x, 1.40 NA, oil). The excitation wavelength was 405 nm. Emission was monitored at 425-550 nm using a hybrid single-molecule detector in photon-counting mode. Each section was recorded as a z-stack with a 2.5 pm step size, covering the full thickness of the section. Maximum-intensity projections were generated from the z-stacks and used for further analysis. Images were analyzed using Fiji software (Schindelin 2012). For measurement of mean fluorescence intensity, images were

[0609]

[0610] manually segmented, excluding background from the analysis. In the oils used as controls (jojoba, almond), the concentration of curcumin was 10 pg / ml and the applied volume was 150 pl.

[0611] Example 92 Determination of Active Substance Penetration into Mucosa and Eye, General ProcedureEyes and mucosal tissues (oral, intestinal, and nasal) were obtained from domestic pigs and sourced from a slaughterhouse (Bocus, Letohrad, Czech Republic). All experiments involving eyes (treatment and rinsing) were conducted at 32 °C. The eyes were either treated and scanned whole, or only histological corneal sections were scanned. If only corneal sections were scanned, the cornea was excised from the eye with a sharp scalpel after the experiment was completed and subsequently frozen in OCT (optimal cutting temperature) medium. Frozen samples were sectioned into 10 pm slices using a Leica CM1950 cryostat. All samples were imaged using a confocal microscope (Leica TCS SP8 X) equipped with an HC PL APO CS2 63x / 1.20 WATER objective. The excitation wavelength was 405 nm. Emission was monitored at 425-550 nm using a hybrid single-molecule detector in photon-counting mode. When whole eyes were scanned, they were imaged in z-stacks reaching a depth of 60 pm with a step size of 0.5 pm. Pixel size was 0.12 pm. Simultaneously with fluorescence, a reflectance image was collected using a 670 nm laser beam and a separate detector. When corneal sections were imaged, they were scanned in z-stacks covering the full thickness of the sample with 2 pm steps. Pixel size was 0.06 pm. The duration and procedure of eye treatment varied between experiments and are described below.

[0612] Experiments with mucosal tissues (treatment and rinsing) were carried out at 37 °C. Mucosal samples were either treated whole and their surface was imaged, or histological sections were scanned. If sections were scanned, the mucosal tissue was frozen in OCT (optimal cutting temperature) medium after the experiment was completed. Frozen samples were sectioned into 10 pm slices using a Leica CM1950 cryostat. All samples were imaged using a confocal microscope (Leica TCS SP8 X). For imaging of sections, an HC PL APO CS2 63x / 1.20 WATER objective was used. The excitation wavelength was 405 nm. Emission was monitored at 425-550 nm using a hybrid single-molecule detector in photon-counting mode. A z-stack covering the full thickness of the sample was acquired with 2 pm steps. For surface imaging of whole tissues, pieces measuring 5 x 5 mm were cut out and placed into wells of an 18-well plate. The wells were filled with 5 ml of treatment or rinsing solution. The entire plate was maintained at 37 °C throughout the experiment. Imaging was performed on a Leica TCS SP8 X confocal microscope (Leica Microsystems) equipped with a Leica HC PL FLUOTAR 5x / 0.15 DRY obj ective. Excitation was provided by a mercury lamp with a FITC filter. Intensity evaluation was performed in Fiji on manually selected 3 x 3 mm regions from the center of the tissue.

[0613] Fresh porcine intestine was cut into pieces approximately 45 cm long. A funnel was inserted into each piece, and each segment was flushed with 2.51 of physiological saline (0.9% NaCl).The intestine was then cut into pieces approximately 15 cm long, which were tied on one end with thread. Subsequently, 10 ml of the solution prepared according to Example 1 was applied into the intestinal lumen. For the control intestine, 10 ml of physiological saline was used. The intestine, tied on both ends, was placed into a 250 ml Duran bottle filled with physiological saline preheated to 37 °C. The bottles were placed on a shaker at 37 °C and 180 rpm for 3 hours. After the experiment, the intestine was cut open and rinsed twice in 250 ml of physiological saline. Tissue pieces were then excised and imaged using a confocal fluorescence microscope (Leica TCS SP8 X) with an HC PL APO CS2 63x / 1.20 WATER objective. Samples were scanned in z-stacks reaching a depth of 90 pm with a step size of 0.36 pm. Pixel size was 0.12 pm. Intensity evaluation was performed in Fiji on manually selected regions of the images; for XZ-plane images, the evaluated region covered the full width of the image and extended to a depth of 50 pm.

[0614] Example 93 Determination of Antioxidant Activity Using the DPPH Process, General Procedure

[0615] Trolox standards at concentrations of 50 to 12.5 pM were used for the measurements and were dissolved in the same manner as the corresponding test samples. Samples containing the HA-C18:1 carrier with active substances (Examples 1, 35, 36, and 38) were diluted 10-fold and tested at a concentration of 100 pg / ml, in comparison with the corresponding active substances alone, controls, and solvents. To 100 pl of each sample solution in a 96-well plate, 100 pl of l,l-diphenyl-2-(2,4,6-trinitrophenyl)hydrazyl (DPPH, c = 0.1 mg / ml) in methanol was added. The decrease in absorbance was measured using a PerkinElmer plate spectrophotometer at a wavelength of 517 nm. Changes in absorbance were expressed as a percentage of the solvent control with DPPH (CTRL). Statistical significance was evaluated using Student’s t-test in Microsoft Excel. Examples 1, 35, 36, and 38 provide the antioxidant activity of the substances in the carriers.

[0616] Example 94 Determination of Antioxidant Activity in 3T3 Cells, General Procedure Antioxidant activity was tested in 3T3 cells seeded in a 96-well plate (5000 cells / well) using the fluorescent probe dichlorodihydrofluorescein diacetate (DCF DA), which is oxidized by hydrogen peroxide, resulting in intense green fluorescence. Prepared carrier samples were diluted 250-fold in serum-free medium. The medium was aspirated from the plate, and the diluted samples were added to the cells as a pretreatment. Each condition was tested in quadruplicate. Incubation was carried out for 3 hours at 37 °C. Meanwhile, a staining solutionof 15 pM DCF DA (prepared by diluting a 1 mM DCF DA / DMSO stock with sterile PBS) was prepared. After the incubation, the treatment solutions were transferred to another plate. The cells were washed with PBS (200 pl / well), followed by addition of the staining solution (200 pl / well). The plate was covered with aluminum foil and incubated for 30 minutes at 37 °C. The cells were then washed twice with PBS (200 pl / well), and the treatment solutions were transferred back. H2O2 was then added to achieve a final concentration of 1.5 mM in each well, followed by a 1-hour incubation at 37 °C. Fluorescence was subsequently measured using a PerkinElmer plate spectrophotometer (excitation 485 nm, emission 535 nm). The fluorescence signal of the samples was compared with controls — cells treated with H2O2. To determine the basal level of DCF fluorescence in 3T3 cells, a negative control (cells not exposed to H2O2) was measured. Results are expressed as % relative ROS concentration compared to the positive control (cells without pretreatment exposed to H2O2). In untreated cells, the ROS level corresponded to 43% of the positive control. Examples 1, 35, 36, and 38 provide the antioxidant activity of the substances in the carriers. Statistical significance was evaluated using Tukey’s test in Origin software.

[0617] Example 95 Determination of Anti-Inflammatory Activity in an Inflammatory Epidermis Model, General Procedure

[0618] Human immortalized keratinocytes (HaCaT) were seeded at a density of 150,000 cells / well in 6-well plates and incubated in 2 ml of medium (with 10% FBS - fetal bovine serum) at 37 °C / 5% CO2, and 95% humidity for 48 h. The cells were then treated with the inflammatory phenotype inducer 250 pM 2-MRC together with the tested potentially anti-inflammatory carrier formulations. Carriers were applied at a concentration of 4 pg / ml (250-fold diluted). Free dexamethasone was used as a positive control for anti-inflammatory activity at a concentration of 1.5 pg / ml. After 24 h, the cells were lysed with RLT buffer (part of the RNeasy Mini Kit, Qiagen), and total RNA was isolated using the QIAcube automated extractor (Qiagen). RNA purity and concentration were verified by absorbance ratios at 260 / 230 nm and 260 / 280 nm (NanoDrop). Gene expression was determined by RT-qPCR. RNA was first reverse-transcribed into cDNA using the High Capacity RNA-to-cDNA Kit (Thermo Fisher Scientific), which served as a template for PCR reactions using TaqMan real-time PCR assays and the StepOne™ Real-Time PCR System (Thermo Fisher Scientific), all according to the manufacturer’s instructions. The probes used included: GAPDH (Hs99999905_m1), HMOX1 (Hs01110250_m1), and IL-1β (Hs01555410_m1) (all from Thermo Fisher Scientific). Obtained CT values were normalized to GAPDH and presented as 2ΔΔCT. Calculated values are presentedas relative gene expression compared to the control (100%). Example 1 reports the relative expression of HM0X1, and Example 38 reports the relative expression of IL-1β after application of the respective carriers. Statistical significance was evaluated using Tukey’s test in Origin software.

[0619] Example 96 Determination of Gene Expression in Porcine Epidermis ex vivo

[0620] Porcine ear samples were obtained from domestic pigs and sourced from a slaughterhouse (Bocus, Letohrad, Czech Republic). For the experiments, skin from the inner part of the auricle was used; it was rinsed, shaved, and wiped with disinfectant. After incubation in culture medium containing a 5-fold higher concentration of antibiotics (penicillin 500U / ml, streptomycin 500 pg / ml) for at least 2h at 4-8 °C, the samples were treated (10-fold diluted and tested at a concentration of 100 pg / ml, in comparison with the corresponding active substances alone, controls, and solvents). In the UV-damage model, the samples were first irradiated with a dose of 80 mJ / cm2using an Oriel UVA+UVB lamp (Newport) and subsequently treated with the test substances. Skin samples were placed on the surface of 0.5% agar in a 24-well plate. After 24 h incubation at 37 °C, the epidermis was removed using the heat-shock procedure (60 °C, 90 s). The epidermis was then homogenized using a Tissue Homogenizer (25-27 Hz; 5 min). Isolation of total RNA, assessment of RNA purity and concentration, and determination of gene expression by RT-qPCR were performed as described in Example 85. The probes used included: MMP-1 (qSscCEP0032832, Bio-Rad), IL-8 (qSscCEP0036027, Bio-Rad), COX-2 (Ss03394694_ml, Thermo Fisher), SOD2 (qSscCEP0043531, Bio-Rad), and ribosomal protein L13a (RPL13A, qSscCEP0038457, Bio-Rad). Obtained CT values were normalized to RPL13 A and presented as 2ΔΔCT. Calculated values are presented as relative gene expression compared to the corresponding control (100%). Statistical significance was evaluated using Tukey’s test in Origin software for IL-8 and COX-2, and using Student’s t-test in Microsoft Excel for MMP-1 and SOD2.

[0621] Example 97 Determination of Cell Migration into a Wound Using the Scratch Assay, General Procedure

[0622] The scratch assay is a procedure that enables monitoring of cell migration into a wound in a 2D environment. HaCaT cells were seeded at a density of 45,000 cells / well into 200 µl of their culture medium (10% FBS, 1.8 mg / ml glucose, 0.3 mg / ml L-glutamine, 100 IU / ml penicillin, 0.1 mg / ml streptomycin, DMEM). The plate was left for 30 min at room temperature in a laminar flow hood and then incubated at 37 °C, 5% CO2. After 24 h of incubation, the culturemedium was replaced with 0% FBS medium (0% FBS, 1.8 mg / ml glucose, 0.3 mg / ml L-glutamine, 100 lU / ml penicillin, 0.1 mg / ml streptomycin, DMEM) to starve the cells prior to the experiment. After another 24 h, the scratch assay was performed using a wound maker (Sartorius). All culture medium was aspirated from the wells, and 100 µl of PBS was added to each well. The wound was created in PBS using the wound maker, after which PBS was aspirated, the wells were washed once with PBS, and treatment solutions (200 µl) were applied. Treatment solutions were diluted either 250-fold or 400-fold in 0% FBS medium. As an untreated control, 0% FBS medium was used, and as a positive control, 10% FBS medium was applied. The plate was placed into the IncuCyte S3 live-cell analysis system (Sartorius). Cells were imaged for 48 h every two hours at 10x magnification in scratch-wound mode. The acquired data were analyzed using IncuCyte 2019B software and expressed as relative cell density within the wound area. Examples 1 and 7 report the efficacy of these dispersed compositions in promoting HaCaT cell migration.

[0623] Example 98 Determination of Antimicrobial Activity, General Procedure

[0624] The sample was applied together with the inoculum onto a Petri dish, and the change in the number of colony-forming units (CFU) per milliliter for selected microorganisms was monitored over time at 25 °C. Example 10 reports the antimicrobial activity of the dispersed composition against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and Aspergillus brasiliensis.

[0625] Example 99 Determination of Curcumin Component Ratio, General Procedure Curcumin was purchased from Sigma-Aldrich. Individual curcumin components were separated using high-performance liquid chromatography (HPLC). Separation was performed on a reverse-phase column with gradient elution. Detection at 425 nm was used to quantify the mass ratio of the individual curcumin components. The studied product contained 77 % curcumin, 17 % demethoxycurcumin, and 6 % bisdem ethoxy curcumin by mass. No significant changes in the ratio of these components were observed.

[0626] Example 100 Lyophilization of Samples and Reconstitution, General Procedure

[0627] A volume of 3.5 ml of the sample was transferred into a clean vial and placed at -20 °C for 2 hours. The vial was then moved to -80 °C and kept there for 0.5 hours. Subsequently, the sample was transferred to a Christ Alpha 2-4 LSC lyophilizer, and the cap was replaced with perforated aluminum foil to allow air flow. Samples were lyophilized overnight (at least16 hours) under the following conditions: condenser temperature -80 °C, pressure 5 x 104Pa. After removal from the lyophilizer, 3.5 ml of demineralized water was added to the lyophilizate, and the solution was stirred for 1 hour on a magnetic stirrer at room temperature.

[0628] Example 101 0.1 mg / ml HA-C18:1 in 55 mM Zn(Ac)2 at pH = 1.13 with 2 mg / ml Curcumin in IPA Prepared by Solvent-Exchange Process, Its Stability and Intestinal Penetration

[0629] Fifty microliters of the dispersion composition prepared according to Example 1 was subjected to a ten-fold dilution with hydrochloric acid (HC1) solution. The ability of the formulation to penetrate intestinal tissue was evaluated 2 hours after dilution according to the procedure described in Example 92.

[0630] pH: 1.13

[0631] Mean particle size by DLS after HC1 dilution: t = 1 h: 1394 nm; t = 2 h: 398 nm

[0632] Stability of encapsulated curcumin in the dispersion composition after HC1 dilution: t = 1 h: 74.2 % of the original mass percentage relative to carrier mass; t = 2 h: 47.9 % of the original mass percentage relative to carrier mass

[0633] Penetration of curcumin into intestinal mucosa: 69.5 photons / pixel (negative control: 15.6 photons / pixel).

[0634] Example 102 0.05 mg / ml HA-C18:1 with 0.05 mg / ml 1.52 x 106g / mol HA in 55 mM Zn(Ac)2at pH = 1.13 with 2 mg / ml Curcumin in IPA Prepared by the Solvent-Exchange Process, Its Stability and Intestinal Penetration

[0635] 50 µl of the dispersion composition prepared according to Example 31 was subjected to a ten-fold dilution with hydrochloric acid (HC1) solution. The ability of the formulation to penetrate intestinal tissue was evaluated 2 hours after dilution according to the procedure described in Example 92.

[0636] pH: 1.13

[0637] Mean particle size by DLS after HC1 dilution: t = 1 h: 603 nm; t = 2 h: 1931 nm

[0638] Stability of encapsulated curcumin in the dispersion composition after HC1 dilution: t = 1 h: 99.5 % of the original mass percentage relative to carrier mass; t = 2 h: 88.7 % of the original mass percentage relative to carrier mass

[0639] Penetration of curcumin into intestinal mucosa: 68.40 photons / pixel (negative control: 15.6 photons / pixel)References:

[0640] Banerjee, S., & Chakravarty, A. R. (2015). Metal complexes of curcumin for cellular imaging, targeting, and photoinduced anticancer activity. Accounts of chemical research, 48(7), 2075-2083.

[0641] Burger, K., Illes, J., Gyurcsik, B., Gazdag, M., Forrai, E., Dekany, I., & Mihalyfi, K. (2001). Metal ion coordination of macromolecular bioligands: formation of zinc (II) complex of hyaluronic acid. Carbohydrate research, 332(2), 197-207.

[0642] Castangia, I., Nacher, A., Caddeo, C., Valenti, D., Fadda, A. M., Diez-Sales, O.,... & Manconi, M. (2014). Fabrication of quercetin and curcumin bionanovesicles for the prevention and rapid regeneration of full-thickness skin defects on mice. Acta biomaterialia, 10(3), 1292-1300. Correa Carvalho, G., Marena, G. D., Gaspar Goncalves Fernandes, M., Ricci Leonardi, G., Santos, H. A., & Chorilli, M. (2024). Curcuma Longa: Nutraceutical Use and Association with Nanotechnology. Advanced healthcare materials, 13(22), 2400506.

[0643] Cowman, M. K., & Turley, E. A. (2023). Functional organization of extracellular hyaluronan, CD44, and RHAMM. Proteoglycan Research, 1(2), e4.

[0644] Čožíková, D., Šílová, T., Moravcová, V., Šmejkalová, D., Pepeliaev, S., Velebný, V., & Hermannová, M. (2017). Preparation and extensive characterization of hyaluronan with narrow molecular weight distribution. Carbohydrate polymers, 160, 134-142.

[0645] Czyzynska-Cichon, I., Janik-Hazuka, M., Szafraniec-Szcz^sny, J., Jasinski, K., W^glarz, W. P, Zapotoczny, S., & Chlopicki, S. (2021). Low dose curcumin administered in hyaluronic acidbased nanocapsules induces hypotensive effect in hypertensive rats. International Journal of Nanomedicine, 1377-1390.

[0646] Davis, B. M., Pahlitzsch, M., Guo, L., Balendra, S., Shah, P., Ravindran, N.,... & Cordeiro, M. F. (2018). Topical curcumin nanocarriers are neuroprotective in eye disease. Scientific reports, 8(1), 11066.

[0647] Eenschooten, C., Vaccaro, A., Delie, F., Guillaumie, F., Tommeraas, K., Kontogeorgis, G. M.,... & Gurny, R. (2012). Novel self-associative and multiphasic nanostructured soft carriers based on amphiphilic hyaluronic acid derivatives. Carbohydrate polymers, 87(1), 444-451.

[0648] European Food Safety Authority (EFSA), Buist, H., Craig, P, Dewhurst, I., Hougaard Bennekou, S., Kneuer, C.,... & Chiusolo, A. (2017). Guidance on dermal absorption. EFSA Journal, 15(6), e04873.

[0649] Fan, Z., Li, J., Liu, J., Jiao, H., & Liu, B. (2018). Anti-inflammation and joint lubrication dual effects of a novel hyaluronic acid / curcumin nanomicelle improve the efficacy of rheumatoid arthritis therapy. ACS applied materials & interfaces, 10(28), 23595-23604.HM Leung, M., Harada, T., & W. Kee, T. (2013). Delivery of curcumin and medicinal effects of the copper (Il)-curcumin complexes. Current Pharmaceutical Design, 19(11), 2070-2083. Hu, J. B., Li, S. J., Kang, X. Q., Qi, J., Wu, J. H., Wang, X. J.,... & Du, Y. Z. (2018). CD44-targeted hyaluronic acid-curcumin prodrug protects renal tubular epithelial cell survival from oxidative stress damage. Carbohydrate polymers, 193, 268-280.

[0650] Huerta-Angeles, G., Bobek, M., Pfikopova, E., Smejkalova, D., & Velebny, V. (2014). Novel synthetic method for the preparation of amphiphilic hyaluronan by means of aliphatic aromatic anhydrides. Carbohydrate polymers, 111, 883-891.

[0651] Huerta- Angeles, G., Brandejsova, M., Kulhanek, J., Pavlik, V., Smejkalova, D., Vagnerova, H., & Velebny, V. (2016). Linolenic acid grafted hyaluronan: Process development, structural characterization, biological assessing, and stability studies. Carbohydrate polymers, 152, 815-824.

[0652] Huerta-Angeles, G., Ondreas, E, Brandejsova, M., Kopecka, K., Vagnerova, H., Kulhanek, J., & Drmota, T. (2020). Formulation of hyaluronan grafted with dodecanoic acid as a potential ophthalmic treatment. Carbohydrate polymers, 246, 116578.

[0653] Hussain, Z., Thu, H. E., Ng, S. E, Khan, S., & Katas, H. (2017). Nanoencapsulation, an efficient and promising approach to maximize wound healing efficacy of curcumin: A review of new trends and state-of-the-art. Colloids and Surfaces B: Biointerfaces, 150, 223-241.

[0654] Hussain, Z., Pandey, M., Choudhury, H., Ying, P. C., Xian, T. M., Kaur, T.,... & Gorain, B. (2020). Hyaluronic acid functionalized nanoparticles for simultaneous delivery of curcumin and resveratrol for management of chronic diabetic wounds: Fabrication, characterization, stability and in vitro release kinetics. Journal of Drug Delivery Science and Technology, 57, 101747.

[0655] Ji, P., Wang, L., Chen, Y, Wang, S., Wu, Z., & Qi, X. (2020). Hyaluronic acid hydrophilic surface rehabilitating curcumin nanocrystals for targeted breast cancer treatment with prolonged biodistribution. Biomaterials science, 8(1), 462-472.

[0656] Kunnumakkara, A. B., Bordoloi, D., Padmavathi, G., Monisha, J., Roy, N. K., Prasad, S., & Aggarwal, B. B. (2017). Curcumin, the golden nutraceutical: multitargeting for multiple chronic diseases. British journal of pharmacology, 174(11), 1325-1348.

[0657] Li, J., Hwang, I. C., Chen, X., & Park, H. J. (2016). Effects of chitosan coating on curcumin loaded nano-emulsion: Study on stability and in vitro digestibility. Food Hydrocolloids, 60, 138-147.Lin, Y, Liu, H., Bu, L., Chen, C., & Ye, X. (2022). Review of the effects and mechanism of curcumin in the treatment of inflammatory bowel disease. Frontiers in Pharmacology, 13, 908077.

[0658] Liu, J., Liu, S., Wu, Y, Xu, X., Li, Q., Yang, M.,... & Du, F. (2022). Curcumin doped zeolitic imidazolate framework nanoplatforms as multifunctional nanocarriers for tumor chemo / immunotherapy. Biomaterials Science, 10(9), 2384-2393.

[0659] Manca, M. L., Castangia, I., Zaru, M., Nacher, A., Valenti, D., Femandez-Busquets, X.,... & Manconi, M. (2015). Development of curcumin loaded sodium hyaluronate immobilized vesicles (hyalurosomes) and their potential on skin inflammation and wound restoring. Biomaterials, 71, 100-109.

[0660] Nesporova, K., Sogorkova, J., Smejkalova, D., Kulhanek, J., Huerta-Angeles, G., Kubala, L., & Velebny, V. (2016). Influence of serum albumin on intracellular delivery of drug-loaded hyaluronan polymeric micelles. International Journal of Pharmaceutics, 511(1), 638-647. Ondreas, E, Dusankova, M., Sita, J., Cepa, M., Stepan, J., Belsky, P, & Velebny, V. (2021). Self-assembly of hydrophobically modified hyaluronic acid. Applied Surface Science, 546, 149161.

[0661] Panahi, Y, Fazlolahzadeh, O., Atkin, S. L., Majeed, M., Butler, A. E., Johnston, T. P., & Sahebkar, A. (2019). Evidence of curcumin and curcumin analogue effects in skin diseases: A narrative review. Journal of cellular physiology, 234(2), 1165-1178.

[0662] Peng, Y, Ao, M., Dong, B., Jiang, Y, Yu, L., Chen, Z.,... & Xu, R. (2021). Anti-inflammatory effects of curcumin in the inflammatory diseases: status, limitations and countermeasures. Drug design, development and therapy, 4503-4525.

[0663] Prasad, S., & Lall, R. (2022). Zinc-curcumin based complexes in health and diseases: An approach in chemopreventive and therapeutic improvement. Journal of Trace Elements in Medicine and Biology, 73, 127023.

[0664] Saso, L., Reza, A., Ng, E., Nguyen, K., Lin, S., Zhang, P,... & Cirillo, N. (2022). A comprehensive analysis of the role of oxidative stress in the pathogenesis and chemoprevention of oral submucous fibrosis. Antioxidants, 11(5), 868.

[0665] Schindelin, J., Arganda-Carreras, I., Frise, E., Kaynig, V, Longair, M., Pietzsch, T.,... & Cardona, A. (2012). Fiji: an open-source platform for biological-image analysis. Nature methods, 9(7), 676-682.

[0666] Shahcheraghi, S. H., Salemi, E, Peirovi, N., Ayatollahi, J., Alam, W., Khan, H., & Saso, L. (2021). Nrf2 regulation by curcumin: molecular aspects for therapeutic prospects. Molecules, 27(1), 167.Starigazdova, J., Nesporova, K., Cepa, M., Sinova, R., Smejkalova, D., Huerta-Angeles, G., & Velebny, V. (2020). In vitro investigation of hyaluronan-based polymeric micelles for drug delivery into the skin: The internalization pathway. European Journal of Pharmaceutical Sciences, 143, 105168.

[0667] Smejkalova, D., Nesporova, K., Hermannova, M., Huerta-Angeles, G., Cozikova, D., Vistejnova, L.,... & Velebny, V. (2014). Paclitaxel isomerisation in polymeric micelles based on hydrophobized hyaluronic acid. International Journal of Pharmaceutics, 466(1-2), 147-155. Smejkalova, D., Muthny, T., Nesporova, K., Hermannova, M., Achbergerova, E., Huerta-Angeles, G.,... & Velebny, V. (2017). Hyaluronan polymeric micelles for topical drug delivery. Carbohydrate Polymers, 156, 86-96.

[0668] Witting, M., Boreham, A., Brodwolf, R., Vavrova, K., Alexiev, U., Friess, W., & Hedtrich, S. (2015). Interactions of hyaluronic acid with the skin and implications for the dermal delivery of biomacromolecules. Molecular pharmaceutics, 12(5), 1391-1401.

[0669] Wu, C. E, Wu, H. J., Shih, C. L., Yeh, T. P, & Ma, W. F. (2024). Efficacy of turmeric in the treatment of oral mucositis in patients with head and neck cancer after radiotherapy or chemoradiotherapy: a systematic review and meta-analysis. Frontiers in Pharmacology, 15, 1363202.

[0670] Zhang, S., Wang, J., Liu, L., Sun, X., Zhou, Y, Chen, S.,... & Li, X. (2022). Efficacy and safety of curcumin in psoriasis: preclinical and clinical evidence and possible mechanisms. Frontiers in Pharmacology, 13, 903160.

[0671] Zhang, Y, Xia, Q., Li, Y, He, Z., Li, Z., Guo, T.,... & Feng, N. (2019). CD44 assists the topical anti-psoriatic efficacy of curcumin-loaded hyaluronan-modified ethosomes: a new strategy for clustering drug in inflammatory skin. Theranostics, 9(1), 48.

Claims

1. C LAIM S1. Dispersion composition, characterized in that it comprises at least one hydrophobically modified hyaluronan derivative of the general formula I as a carrier,NHwhere,R is H+or a physiologically acceptable metal cation selected from any alkali metal ions or alkaline-earth metal ions or zinc, preferably Na+, K+, Mg2+, Ca2+, or Zn2+,R1is -H or an acyl group -C(=O)CxHy, where x is an integer from 6 to 22 and y is an integer from 14 to 46, and CxHyis a linear or branched, saturated or unsaturated chain, wherein there is at least one acyl group in at least one repeating unit of the modified derivative,with the proviso thatwhen R1is a Ce to Cio acyl, n is an integer in the range from 13 to 2500, preferably from 20 to 1500, with a degree of substitution of 5 to 70 %, preferably 8 to 60 %.with the further proviso that when R1is a Cn to Ci6 acyl, n is an integer in the range from 13 to 500 with a degree of substitution of 5 to 20 %, or n is an integer in the range from 500 to 1750 with a degree of substitution of 4 to 12 %,with the further proviso that when R1is a C17 to C22 acyl, n is an integer in the range from 13 to 500, preferably from 20 to 250, with a degree of substitution of 3 to 15 %, preferably 4 to 10 %;wherein the concentration of the hydrophobically modified hyaluronan of general formula I is in the range of 0.03 to 0.4 wt.% relative to the total weight of the composition;and wherein the composition further comprises zinc acetate at a concentration in the range of 0.4 to 5 wt.% relative to the total weight of the composition,and further comprises encapsulated curcumin of the general formula II as the primary active substance.(II),where R2is -H or -O-CH3, or a mixture of curcumins, wherein the curcumin is selected from the group comprising [(lE^E)- l,7-bis(4-hydroxy-3-methoxyphenyl)-l,6-heptadiene-3, 5-dione], demethoxycurcumin, or bisdemethoxycurcumin;wherein the concentration of curcumin of general formula II or mixtures thereof is in the range of 0.003 to 0.1 wt. % relative to the total weight of the composition; and water or an aqueous salt solution selected from the group comprising aqueous solutions of chlorides or acetates of physiologically acceptable metals, wherein the physiologically acceptable metals are selected from the group comprising alkali metal ions, alkaline-earth metal ions, or zinc;wherein the stability of the encapsulated curcumin in the dispersion composition is characterized in that at least 8 wt. % of the encapsulated curcumin relative to the weight of the carrier, and at least 80 % of the original amount of encapsulated curcumin is retained for at least 3 months at 40 °C, or at least 90 % of the original amount of encapsulated curcumin is retained for at least 18 months at room temperature, or at least 90 % of the original amount of encapsulated curcumin is retained for at least 18 months at 5 °C.

2. The composition according to claim 1, characterized in that R1is oleoyl, wherein the weight- average molar mass of oleoyl hyaluronan is in the range of 5 x 103to 3 x 104g mol-1, with a degree of substitution in the range of 3 to 15 %, preferably 1 x 104to 2 x 104g mol-1with a degree of substitution in the range of 5 to 10 %.

3. The composition according to any one of claims 1 to 2, characterized in that it further comprises a secondary active substance or a combination thereof selected from the group of hydrophilic active substances comprising native hyaluronan, chondroitin sulfate, glucosamine sulfate, metformin, heparin, inositol, vitamin C and B-vitamingroup, lactic acid, amino acids, a-hydroxy and P-hydroxy acids, peptides, water-soluble proteins, carboxymethyl glucan, schizophyllan, glucomannan, panthenol, urea, glycerol, pentylene glycol, preferably native hyaluronan, and / or from the group of hydrophobic active substances comprising dexamethasone, resveratrol, or cannabidiol, preferably dexamethasone, wherein the concentration of the secondary hydrophilic active substance or combination thereof is 0.0001 to 3 wt. % and / or the concentration of the secondary hydrophobic active substance is 0.001 to 0.1 wt. %.

4. The composition according to claim 3, characterized in that it comprises native hyaluronan having a weight average molar mass in the range of 5 x 103to 2 x 106g.moT preferably in the range of 1 x 104to 1.5 x 106g moT1.

5. The composition according to any one of claims 1 to 4, characterized in that it further comprises an excipient or a combination thereof selected from the group comprising viscosity regulators, preferably xanthan gum, guar gum, carboxymethylcellulose, sodium alginate, pectin, carbomer, polyethylene glycol, polyvinylpyrrolidone, hydrolyzed collagen, gelatin, more preferably xanthan gum; preservatives, preferably aromatic acids and their derivatives, more preferably benzoic acid, salicylic acid, dehydroacetic acid, potassium sorbate, parabens; alcohols, preferably ethanol, isopropanol, benzyl alcohol, phenoxyethanol, phenethyl alcohol, sorbitol, pentylene glycol; sugars, preferably sucrose, glucose, fructose, mannose, galactose; salts, preferably sodium chloride, wherein the concentration of the excipient or combination thereof is 0.001 to 5 wt. %.

6. A method for preparing the composition defined in any one of claims 1 to 5, characterized in that curcumin dissolved in an organic solvent is added to an aqueous solution of at least one hydrophobically modified hyaluronan derivative of general formula I and zinc acetate, after which the organic solvent is evaporated, and the non-encapsulated curcumin is subsequently removed from the mixture, preferably by filtration, more preferably through a 1 μm glass filter, wherein the organic solvent is selected from the group consisting of acetonitrile, ethanol, and isopropanol, preferably isopropanol.

7. The method of preparation according to claim 6, characterized in that the concentration of the hydrophobically modified hyaluronan is in the range of 0.3 to 4 g / 1, preferably 0.5 to 2 g / 1, more preferably 1 g / 1.

8. The method of preparation according to claim 6 or claim 7, characterized in that the concentration of zinc acetate is in the range of 25 to 200 mM, preferably 30 to 70 mM, more preferably 55 mM.

9. The method of preparation according to any one of claims 6 to 8, characterized in that the concentration of curcumin in the organic solvent is in the range of 0.5 to 5 g / 1, preferably 2 to 4 g / 1.

10. The method of preparation according to any one of claims 6 to 9, characterized in that at least one secondary hydrophilic active substance dissolved in an aqueous solution is added, preferably in an aqueous zinc acetate solution having a concentration of 30 to 70 mM.

11. The method of preparation according to any one of claims 6 to 10, characterized in that at least one secondary hydrophobic active substance dissolved at a concentration of 0.5 to 5 g / 1, preferably 2 to 4 g / 1, in an organic solvent is added, after which the organic solvent is evaporated and the non-encapsulated hydrophobic active substance is removed, preferably by filtration, more preferably through a 1 μm glass filter.

12. The method of preparation according to any one of claims 6 to 11, characterized in that the volume of the prepared dispersion composition, after evaporation of the organic solvent, is adjusted with water or an aqueous salt solution selected from the group containing aqueous solutions of chlorides or acetates of physiologically acceptable metals, wherein the physiologically acceptable metal ions are selected from the group containing alkali metal ions, alkaline-earth metal ions, or zinc ions.

13. The method of preparation according to any one of claims 6 to 12, characterized in that the composition is subsequently converted into a dry form, preferably by lyophilization.

14. A pharmaceutical or cosmetic composition, characterized in that it comprises the dispersion composition of any one of claims 1 to 5.

15. The composition according to claim 14, characterized in that it is in a form selected from the group comprising a cream, gel, emulsion, serum, or hydrogel.

16. The composition according to claim 15, characterized in that it is in the form of an emulsion, wherein the concentration of the dispersion composition is in the range of 0.001 to 99.999 wt. / wt.%, preferably 0.01 to 10 wt. / wt.%, more preferably 0.1 to 5 wt. / wt.%.

17. The composition according to claim 15, characterized in that it is in the form of a serum, wherein the concentration of the dispersion composition is in the range of 0.001 to 100 wt. / wt.%, preferably 0.01 to 10 wt. / wt.%, more preferably 0.1 to 5 wt. / wt.%.

18. The composition according to claim 15, characterized in that it is in the form of a cream, wherein the concentration of the dispersion composition is in the range of 0.001 to 50 wt. / wt.%, preferably 0.01 to 10 wt. / wt.%, more preferably 0.1 to 5 wt. / wt.%.

19. The composition according to claim 15, characterized in that it is in the form of a gel or hydrogel, wherein the concentration of the dispersion composition is in the range of 0.001 to 100 wt. / wt.%, preferably 0.01 to 95 wt. / wt.%, more preferably 0.1 to 10 wt. / wt.%.

20. The composition according to any one of claims 15 to 19, characterized in that it comprises the dispersion composition and at least one cosmetic or pharmaceutical excipient selected from the group comprising an oil, wax, butter, emulsifier, secondary active substance, viscosity regulator, and preservative.

21. A nutraceutical composition, characterized in that it comprises the dispersion composition of any one of claims 1 to 5.

22. The nutraceutical composition according to claim 21, characterized in that it is in a form selected from the group comprising a gel, emulsion, solution, syrup, jelly, or hydrogel.

23. The dispersion composition according to any one of claims 1 to 5; or the composition according to any one of claims 14 to 20, for use in a pharmaceutical application, preferably for treatment of skin or mucosa, or for topical application, or for ophthalmic application.

24. The dispersion composition according to any one of claims 1 to 5; or the composition according to claim 21 or claim 22, for use in oral administration, preferably in the form of a dietary supplement.

25. Use of the dispersion composition according to any one of claims 1 to 5, or the composition according to any one of claims 14 to 20, for a cosmetic application, preferably for cosmetic treatment of the skin.

Images