WO2024214063A1

WO2024214063A1 - Method for preservation of an active compound, formulation and preservative

Info

Publication number: WO2024214063A1
Application number: PCT/IB2024/053608
Authority: WO
Inventors: Tom KAMPERMAN; Sara TAHAN LATIBARI; Jenny Evelin PARRAGA MENESES; Barbara Maria ZOETEBIER
Original assignee: Iamfluidics Holding BV
Current assignee: Iamfluidics Holding BV
Priority date: 2023-04-14
Filing date: 2024-04-12
Publication date: 2024-10-17
Anticipated expiration: 2025-10-14
Also published as: NL2034584B1; CN121152674A

Abstract

In a method and preservative for preservation of an active compound, said compound is provided as a powder comprising said compound. Said powder is dispersed in an oleogel to form a suspension. Said suspension, containing said powder in an oleogel network or matrix, is formed into micro-bodies and said micro-bodies are encapsulated by a hydrophilic polymer shell to form micro-bodies containing said active compound.

Description

Method for preservation of an active compound, formulation and preservative

The present invention relates to a method for preservation of an active compound, as well to a preservative and a formulation.

Many active compounds are prone to deterioration when exposed to an oxidizing or hydrating environment like the surrounding air. Particularly a hygroscopic compound may attract moisture and become sticky or even become a viscous gel or paste. This may impede handling the compound and may even increase and/or accelerate its deterioration. Attempts have been made to protect the active compound against such deterioration. A common approach is to contain the product in an air tight package that was vacuumed and/or flushed with an inert or protecting gas. Opening such package, however, will again expose the product to the surrounding air and therefore will diminish its shelf life.

A more sophisticated method for preservation of an active compound is described in International patent application WO2022/029623 by applicant. This application describes a process by which a reactive compound is dissolved in a suitable hydrophilic solvent. The obtained solution is emulsified with a hydrophobic liquid and formed into micro-droplets that are encapsulated with a hydrophilic polymer shell to form micro-capsules. Such preservation has proved to extend the lifetime of the active ingredient considerably.

It is an object of the present invention to improve the preservation of a reactive compound even further by providing an alternative preservation process and preservative.

To that end the present invention provides a method for preservation of an active compound, wherein a powder of said active compound is provided, wherein said powder is dispersed in an at least partly gellable or crystallizable liquid hydrophobic material to form a suspension or dispersion containing said powder, wherein said suspension or dispersion is formed into micro-bodies, wherein said micro-bodies are encapsulated by a polymer shell to form suspension- or dispersion-laden micro-bodies which contain one or multiple cores of said suspension or dispersion, wherein said suspension- or dispersion-laden micro-bodies are solidified, particularly gelled and/or crystallized, to form micro-bodies containing said active compound in an at least substantially solidified hydrophobic network.

It is noted that when used in the present application, unless explicitly stated otherwise, the expression "micro-bodies" can also be referred to interchangeably as "microdroplets", "micro-capsules" or "micro-particles", while the adjective "micro" refers to sizes ranging from a sub-micron, particularly nano, scale to several millimetres. The descriptor "(partly or completely) solidified, particularly gelled and/or crystallized, hydrophobic network" can also be referred to as "oleogel" and vice versa. Oleogels, sometimes also called organo-gels, are gels having a continuous phase of oil. Oleogel networks are characterized by their capacity to retain substantial amounts of oil. Oleogels are further characterized by their thermo-reversible, viscoelastic, and (under certain conditions) substantially self-standing properties. The oleogel may be applied in combination with a hydrogel, also referred to as a bi-gel.

Also, unless explicitly stated otherwise, the adjective "micro" is meant to denote dimensions up to a few millimetre and below. Micro-bodies, accordingly, can have a diameter or maximum dimension ranging from several micrometre (or micron), particularly several tens of micrometre, particularly a few hundred micrometre, up to a few millimetre. Due to the relative small dimension of the micro-bodies, the active compound may still readily be used as a powdery or granular ingredient in a variety of applications, formulations, and products. The powder comprising the active compound is captured inside these water-free micro-bodies that protect the compound against the external environment.

In a further aspect of the invention, a preservative for an active compound, according to the invention, comprises one or more preserving micro-bodies, each having at least one core surrounded by a shell that is formed by a polymer network, wherein said core comprises an at least substantially solidified, particularly gelled, gellable or crystallizable liquid hydrophobic material, particularly an oleogel, and a solid powder, said powder being dispersed throughout said core, and wherein said powder comprises said active compound.

In a further aspect, the invention provides for a formulation comprising an active compound in a preserved state by means of such preservative. Such formulation may be applied in or as a cosmetic product, a personal care or home care product, a dermatological product, a food or nutrition product, an agrochemical product, a fragrance product, a health or healthy product, a pharmaceutical product, a (bio)medical product, a household product, an energy storage product, a coatings product, or an adhesives product.

As a result, the active compound is dispersed and captured inside a substantially water-free, hydrophobic and air-tight environment provided by the oleogel surroundings of said core. This appears to greatly enhance the shelf-life of the compound concerned. The core also appears to substantially prevent both the release and dissolution of the powder into aqueous and/or hydrophilic solvents, formulations or surroundings with which the micro-bodies may be mixed. The polymer shell holds the core together and avoids deformation, merging, or damaging of the oleogel cores and individual micro-bodies.

Furthermore, the polymer shell enables facile handling such as drying, mixing and/or blending of the hydrophobic oleogel into other formulations such as cosmetic, food, and pharmaceutical formulations, including non-oil-based and hydrophilic, for example, water-based formulations. The polymer shell may be designed to disintegrate or break under induced chemical or mechanical conditions, upon which said active compound comprising powder is released from the micro-body. The active compound is, as it was, entrapped and substantially hermetically canned inside a micro-body until being released. The active compound can be released via mechanical and chemical conditions or triggers. Such induced conditions or triggers, may for instance include shear stress, like for instance rubbing on the skin or brushing, for example by means of a paint brush, compression, like chewing, a combination of shear and compression, for instance extrusion or injection through a small dispenser like a needle or spray nozzle, disintegration of the core or the shell, for instance by chelation, freeze-thaw cycles, or heating, particularly to promote oleogel melting, hydrolyzation, photodegradation, diffusion, burning, acidic or enzymatic degradation or fermentation, for instance in the stomach, ileum, colon, or any other part of the gastrointestinal tract, or any other form of mechanical and/or chemical stress that results in the escape of the active compound from a micro-body. The micro-bodies may be exposed to a solution comprising citric acid, where said citric acid concentration is between 0.01 wt% and 100 wt%, particularly between 0.1 wt% and 10 wt%, particularly between 0.5 wt% and 5 wt%.

In a specific embodiment, the method, formulation and preservative according to the invention are characterized in that said shell comprises a polymer network, particularly an interpenetrating network, double network, and/or composite of two or more inter- or intra- cross-linked polymers, and more particularly in that said polymer network comprises a hydrophilic polymer network, particularly comprising one or more poly-electrolytes or polysaccharides selected from agar, alginate, chitosan, dextran, polyjethylene glycol), collagen, gelatin, hyaluronic acid, carrageenan, particularly Lambda-, Kappa and Lota carrageenan, fibroin, fibronectin, poly-l-lysine (PLL), cellulose, graphene, poly(ethylenimine) (PEI), poly(amidoamine) (PAA), dextran sulfate, silk, silk fibroin, pectin, locust bean gum, gellan gum, guar gum, tragacanth gum, xanthan gum, acacia gum, karaya gum, starch, and sodium carboxymethyl cellulose (S-CMC), all of these as naturally derived materials and/or synthetically derived materials including recombinant proteins and/or derivatives of these materials.

In further embodiment, the method, formulation and preservative according to the invention are characterized in that said shell comprises a bio-compatible, biodegradable and/or bio-resorbable polymer network, particularly a polymer network comprising a methyl-methacrylate derivative, a caprolactone derivative, a lactic acid derivative, a glycolic acid derivative, and/or a co-polymer of lactic acid and glycolic acid, more particularly in that said polymer network comprises a poly(lactic-co-glycolic acid), a poly(caprolactone) and/or a poly(methyl-methacrylate).

Particularly successful results were obtained in this respect with a further specific embodiment of the method, formulation and preservative according to the invention, wherein said polymer network comprises a cross-linked or inter-penetrating alginate network, particularly a calcium cross-linked alginate network. The network may be further strengthened by incorporation of nano- or micro-particles and/or polyelectrolytes.

In a specific embodiment the method, formulation and preservative according to the invention are characterized in that said polymer shell comprises an alginate network, particularly a sodium-, calcium- and/or a shellac reinforced alginate network.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said liquid hydrophobic material comprises at least one oil, particularly one or more oils from a group comprising: vegetable oils, such as sunflower oil, corn oil, linseed oil, canola oil, castor oil, palm oil, coconut oil, avocado oil, sweet almond oil, calophylum oil, sesame oil, olive oil, jojoba oil, soybean oil, cottonseed oil, rapeseed oil, peanut oil, flaxseed oil, borage oil, safflower oil, macro-algae oil and seaweed oil; essential oils, ethereal oils, macerated oils, triglyceride, animal oils, such as tallow fat, lanolin and marine oils, such as fish oils; synthetic oils, and neutral oils like medium chain triglyceride oils, as well as mixtures or derivatives thereof, and more particularly oils that are non-toxic and user-friendly, particularly sunflower oil or safflower oil. Particularly, the method, preservative or formulation according to the invention may be characterized in that said liquid hydrophobic material comprises sunflower oil or safflower oil. More particularly, the method, preservative or formulation according to the invention may be characterized in that said liquid hydrophobic material is an oleogel comprising an oil, particularly sunflower oil or safflower oil.

In a further particularly embodiment, the method, preservative or formulation according to the invention is characterized in that said liquid hydrophobic material is an oleogel comprising an oil, more particularly comprising an oil with an oleic acid content higher than 10%, more particularly higher than 25%, more particularly higher than 50%, more particularly higher than 70%, even more particularly comprising high-oleic sunflower oil characterized by an oleic acid content higher than 70%.

The oleogel may comprise an oil gelling agent at a concentration of at least 1 wt% (gelling agent to oleogel weight percentage), particularly at least 2 wt%, particularly at least 3 wt%, particularly at least 4 wt%, particularly at least 5 wt%, particularly between 1 wt% and 10 wt%, particularly between 4 wt% and 6 wt%.

Particularly, the method, preservative or formulation according to the invention may be characterized in that said oleogel retains substantial amounts of said oil, more particularly wherein said oleogel has the capacity to bind or retain more than 90% of said oil in said oleogel, particularly more than 95%, during at least 4 weeks at at least room temperature, particularly at 40 degrees Celsius.

The capacity of said oleogel to bind said oil is also referred to as 'oil binding capacity¹ and is equal to the weight of bound oil determined after, for example, centrifugating the oil/oleogel mixture, divided by the weight of solid fat determined by, for example, pulse nuclear magnetic resonance (NMR). Alternatively, stability of said oil binding capacity may be determined by letting separate the unbound oil phase from said oleogel over time and determining the weight and/or volume ratio(s) of unbound oil and oleogel. For various markets, ethereal, macerated and/or essential oils or waxes furthermore add favourable, pleasant organoleptic properties or therapeutic benefits to the product. Examples of suitable organic lipophilic compounds are for instance: immortelle, lavender, german chamomile, neroli, peppermint oil, rosemary, rose oil, tea tree oil, dwarf pine, juniper berry, roast chestnut extract, birch leaf extract, hayseed extract, ethyl acetate, camphor, menthol, rosemary extract, eucalyptus oil, cranberry oil.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said liquid hydrophobic material comprises at least one hydrophobic gelling agent, specifically a fatty acid, a wax, or a sterol that is provided in liquid form and that is gelled, crystallized or otherwise solidified into said micro-bodies. Accordingly the liquid material is processed in a liquid state and allowed to solidify below a gelling temperature of the hydrophobic gelling agent to form crystals incorporated in a hydrophobic network, also referred to as oleogel. The suspended powder will become captured (trapped) in such stable solidified oleogel matrix that prevents migration and/or phase separation of said suspension.

In this respect, the liquid hydrophobic material may particularly comprise one or more fatty acids and/or waxes from a group comprising: paraffin wax, rice bran wax, sunflower wax, carnauba wax, candelilla wax, beeswax, microcrystalline wax, coconut wax, ozocerite wax, beta-sitosterol, gamma-oryzanol, stearic acid, palmitic acid, behenic acid, myristic acid, lauric acid, capric acid, and fatty acid derivatives, such as fatty acid esters with short chain alcohols, such as isopropyl myristate, isopropyl palmitate and isopropyl stearate and dibutyl adipate, and particularly carnuba wax.

It is noted that the gelling, gelation, melting or crystallization temperature of the oleogel and/or hydrogel, all collectively and separately also referred to as solidification temperature, may typically be not a single point, but may occur over a temperature range characterized by an onset and an offset temperature, or specifically a melting onset and melting offset temperature, a gelling onset and gelling offset temperature, or a crystallization onset and crystallization offset temperature, wherein 'onset' refers to starting and 'offset' refers to completion of the respective process. Also note that the solidification temperature of the liquid material can be lower than the melting/crystallization temperature, in other words, the crystallization offset temperature can be lower than the crystallization onset temperature. It is also noted that the expressions "gelation" and "solidification" and like derivatives, such as "gelled", "gelling" and "solidified", may be used interchangeably.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said oleogel comprises a hydrophilic gelling agent, in particular a polysaccharide, a (modified) starch, a protein, a natural gum, a hydrocolloid, a steroid, a hydrophilic phytosterol derivative, or a phospholipid, in particular one or more gelling agents selected from the group of cellulose, maltodextrin, dextran, hyaluronic acid, gelatin, whey protein, gum Arabic, gum tragacanth, xanthan gum, carrageenan, agar, lecithin, propylene glycol, silica (nanoparticles), more particular ethyl cellulose. The hydrophilic gelling agent aids in further stabilizing the suspension of said powder and said oleogel.

In general, fats, waxes, and oleogels are solid or creamy (malleable) at room temperature. Advantage may be taken of this natural nature of fats, waxes, and oleogels to further immobilize the active compound within the capsule. Accordingly, a further particular embodiment of the method according to the invention is characterized in that said liquid hydrophobic material is processed in a liquid condition to at least partly solidify or have solidified below a solidification temperature thereof. In a further specific embodiment the method according to the invention is characterized in that said liquid hydrophobic material is cooled down to below said solidification temperature thereof at a cooling rate faster than 0.1 K/min, particularly faster than 1 K/min, more particularly faster than 10 K/min, and more particularly faster than 100 K/min.

The solidification temperature of the liquid material can be tuned by changing its composition, particularly the types and concentrations of oils, waxes, and other components in an oleogel. These wax or fat containing oleogels may be processed in a liquid form to create the micro-bodies or capsules, particularly by using the method of co-pending European patent application by the same applicant that published as EP 3.436.188 Al and whose subject matter is herewith incorporated by reference.

Subsequently the product may be stored below the oleogel gelation temperature, which could, depending on the oleogel composition, for example be at 40 degrees Celsius, room temperature, or at 4 degrees Celsius. While the oleogel contained in the capsule is in a solid state it will counteract migration of both the active compound out of the capsule as well as of environmental compounds into the capsule. Storing the micro-bodies at temperatures below the solidification temperature of the oleogel may also lead to a denser oleogel network and further strengthening of the oleogel network in time.

The oleogel composition according to the present invention may be such that the oleogel solidification temperature is between 0 and 100 degrees Celsius, particularly between 4 and 90 degrees Celsius, particularly between 4 and 40 degrees Celsius, particularly between 20 and 40 degrees Celsius, particularly between 20 and 90 degrees Celsius, particularly between 40 and 90 degrees Celsius, particularly between 40 and 50 degrees Celsius, particularly between 50 and 60 degrees Celsius, particularly between 60 and 70 degrees Celsius, particularly between 80 and 90 degrees Celsius, and preferably above room temperature.

Preferably said liquid form condition is attained without adversely affecting the integrity of the active compound. To that end, a further embodiment of the method, formulation and preservative according to the invention have the feature that said oleogel has a solidification offset temperature below about 60 degrees Celsius. This suppresses thermally induced degradation of the active compound. Suitable candidates to formulate said oleogels are for instance a wax that is selected from a group of montan wax, carnauba wax, glycol montanate, paraffin wax, rice bran wax, sunflower wax, candelilla wax, beeswax, microcrystalline wax, coconut wax, ozocerite wax and mixtures thereof.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said oleogel comprises an emulsifier or plasticizer, such as fatty acids, glycolipids, monoglycerides, diglycerides, triglycerides, or phospholipids, or a mixture thereof, particularly soy lecithin or polyglycerol polyricinoleate (PGPR).

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said oleogel comprises an oil-soluble anti-oxidizing agent, such as, for example, a tocopherol (known as vitamin E), particularly alpha-tocopherol, particularly 0.01-10 wt% alpha-tocopherol in oleogel, particularly 0.1-1% alpha-tocopherol in oleogel.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder is provided as a dry, substantially water-free powder. Removing any initial water or water vapour will increase the lifetime of the powdery ingredient.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises an anti-oxidizing agent and/or a nutrient, particularly one or more from the group of catalase, polyphenol, curcumin, quercetin, catechin, lignan, resveratrol, citric acid, and L-ascorbic acid. In particular said powder comprises L-ascorbic acid. The encapsulation according to the invention protects the compound from oxidative and/or pro-oxidative conditions such as moisture and oxygen from ambient air or from a liquid formulation in which the micro-bodies are mixed, thereby retaining at least a substantial part of its original nutrient and/or anti-oxidation capabilities, particularly as compared to non-encapsulated anti-oxidizing agent and/or nutrients. In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said active compound is a hydrating agent, particularly hyaluronic acid, alginate, gelatin or collagen. The encapsulation according to the invention substantially shields the compound from ambient air and/or from water, thereby preventing degradation of the hydrating agent, for example through hydrolysis, thereby retaining its original structure and hydrating capabilities.

Furthermore, micro-bodies that contain a water-shielding oleogel advantageously allow for the encapsulation of relatively high concentrations of hygroscopic compounds such as hydrophilic polymers, like dry hyaluronic acid powder. In such environment, the hygroscopic compound remains a dry, non-wetted powder that may have been blended within a molten oleogel during the encapsulation process, which enables more facile processing and easier encapsulation of higher polymer concentrations as compared to handling the hygroscopic compound that is not mixed with a oleogel.

Specifically, hydrophilic polymers can become notoriously sticky and viscous upon interaction with water or moisture, especially at relatively high molecular weights (> 100 kDa, >1000 kDa) and/or high water-binding molecules, such as, for example, high molecular weight hyaluronic acid. Sticky powders or high viscous solutions are difficult to process and particularly difficult to encapsulate. In a further specific embodiment the method, formulation and preservative according to the invention are, however, characterized in that said powder comprises a hydrophilic polymer with a molecular weight larger than 10 kDa, particularly larger than 100 kDa, particularly larger than 1000 kDa, in particular a polysaccharide, in particular hyaluronic acid. The present invention enables encapsulation of such relatively high concentrations of hygroscopic compounds in their dry powder form by dispersing them within oleogel and substantially protecting them from contact with water or other aqueous solvents.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises a hygroscopic compound, particularly a natural compound, particularly from the group of hydrophilic polymers, polysaccharides, proteins, nucleic acids, and water-soluble salts.

In another specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises a hygroscopic salt, in particular a salt of a quaternary ammonium cation, more particularly a choline salt, in particular one from the group of choline chloride, choline bitartrate, choline borate, choline dihydrogen citrate, choline bicarbonate and choline chloride carbamate, even more particularly choline chloride.

Choline salts are used in nutrition, for example as adjuncts to animal and poultry feeds. In such instances, the hygroscopic nature of the choline salts that are commonly available makes uniform mixing difficult and the preparation otherwise unsatisfactory. Furthermore, due the hygroscopic nature of most choline salts, the salts tend to liquify, which is problematic for processing and also causes undesirable odor of the material. Mixing dry choline salts in liquified oleogel and subsequently encapsulating in a preserving micro-body according to the invention protects the choline against moisturization, sticking together, and deterioration.

In another specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises a dried, spray dried, or freeze dried composition comprising an active ingredient (hygroscopic) drying excipient, particularly a drying excipient comprising at least one polycarbohydrate, monosaccharide, disaccharide, or polysaccharide compound, and more particularly a drying excipient being a sugar, more particularly the drying excipient being selected from the group of dextran, dextrin, maltodextrin, trehalose, lactose, glucose, dextrose, sucrose, fructose, maltose, isomaltose, sorbitol, mannitol, lactitol, xylitol, and/or erythritol.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises a biological compound, particularly an organism, a protein, an enzyme, a peptide, a nucleotide, or a mixture thereof.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises a viable compound, particularly a living cell, more particularly a bacteria or microbiota.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises one or more sensitive bacteria selected from the group of anaerobic bacteria, non-spore forming bacteria, and other bacteria sensitive to moisture, acid, heat, and/or oxygen.

In a further aspect of the invention said preserving micro-bodies at least partially protects said sensitive bacteria and thereby increase the survival of the bacteria during and/or after the production process and/or extend the shelf life of a product. In other words, said active compound, particularly said sensitive bacteria, are more stable and have a higher viability during and after production of a product if said active compound, particularly said sensitive bacteria are encapsulated in said preserving micro-bodies as compared to said active compound, particularly said sensitive bacteria that are not encapsulated in said preserving micro-bodies.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises one or more bacteria from a group of Bacteroides, Bifidobacterium, Fusobacterium, Bacillus, Lactobacillus, Saccharomyces, Streptococcus, Enterococcus Porphyromonas, Prevotella, Actinomyces, Propionibacterium, Clostridia, particularly one or more from the group of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccharomyces boulardii, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus fermentum, Bacillus coagulans, Bacillus subtilis,

Enterococcus faecium.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises one or more anaerobic bacteria from the group of Bacteroides, Bifidobacterium, Fusobacterium, Porphyromonas, Prevotella, Actinomyces, Propionibacterium, Clostridia, particularly one or more from the group of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium longum.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises one or so called 'next-generation probiotics', more particularly said powder comprises one or more next-generation probiotics from the group of Akkermansia muciniphila, Faecalibacterium prausnitzii, Bacteroides thetaiotaomicron, Bacteroides fragilis, Roseburia spp., Prevotella spp., Alistipes spp., Christensenella minuta, Blautia spp., Eubacterium hallii, and Methanobrevibacter smithii

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises one ore more prebiotics, particularly one ore more from the group of oligosaccharides, fructans, galactans, starch, pectins, and beta-glucans. Prebiotics are also referred to as dietary fibers.

In a further specific embodiment the method according to the invention is characterized in that said liquid hydrophobic material is solidified by combining the micro-bodies with a phase, particularly a liquid phase, that has a temperature lower than a solidification temperature of said liquid hydrophobic material, particularly a temperature below room temperature, more particularly a temperature below 10 degrees Celsius. In a further specific embodiment the method according to the invention is characterized in that said phase comprises a liquid phase containing a cross-linking compound to cross-link said shell polymer. These micro-bodies may be encapsulated by combining a liquid stream containing said cross-linking compound with a stream of said micro-bodies, particularly by using the method of a co-pending European patent application by applicant that published as EP 3.436.188 Al whose subject matter is herewith incorporated by reference.

In a further specific embodiment the method according to the invention are characterized in that said liquid phase has a temperature that induces thermal or physical cross-linking of at least on of said shell polymers.

The micro-bodies may stored in a carrier liquid at a temperature above a freezing temperature of the said carrier liquid and below said oleogel solidification temperature, particularly below room temperature, particularly below 10 degrees Celsius, particularly at approximately 4 degrees Celsius.

In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said powder comprises a catalyst, in particular one or more compounds from a group of compounds that catalyse amino cross-linking, particularly comprising p-TSA (para-Toluene

Sulfonic Acid), DNNDSA (Di Nonyl Naphtalene Di Sulfonic Acid), DDBSA (Do Decyl Benzene Sulfonic Acid), DNNSA (Di Nonyl Naphtalene mono Sulfonic Acid), and Phosphate Acid, and Carboxylic Acid; or catalyze urethane cross-linking, particularly including DBTL (Di Butyl Tin Laurate), DOTL (Di Octyl Tin Laurate), DBTO (Di Butyl Tin Oxide), Bismuth-based catalyst, Zirconium-based catalyst, and amine-based catalyst.

In a further specific embodiment the method according to the invention is characterized in that said one or more micro-bodies are laden with a hydrophobic compound by immersing said one or more micro-bodies into a solution of said hydrophobic compound, particularly a concentrated solution, allowing said hydrophobic compound to diffuse trough said shell into said one or more cores of said one or more micro-bodies.

In a further specific embodiment the method according to the invention is characterized in that said one or more micro-bodies are laden with a hydrophobic compound by providing said one or more micro-bodies with one or more cores in molten state prior to immersing said one or more micro-bodies into a solution of said hydrophobic compound, particularly a concentrated solution, to allow said hydrophobic compound to diffuse into said one or more cores of said one or more micro-bodies in said molten state, and subsequently allowing said one or more cores to at least substantially solidify. The cores may for instance comprise a molten oil or wax. After such core is solidified again, for instance by re/gelling, the hydrophobic compound will be trapped in the micro-bodies.

In a further specific embodiment the method according to the invention is characterized in that said shell comprises a calcium-alginate network that is weakened or completely removed prior to or during final application, particularly by scavenging calcium ions and at least partly disrupting the calcium-alginate network of the shell, more particularly by introducing a chelator agent such as citric acid or ethylene- diamine-tetra-acetic acid (EDTA).

In a further specific embodiment the method according to the invention is characterized in that said micro-bodies are dried, particularly by a drying method chosen from a group containing evaporation, blowing dry gas, vacuum drying, fluid bed drying, freeze drying, microwave drying, and chemical drying. Such method will remove water from the micro-bodies and in particular from the hydrophilic shell. In a further specific embodiment dry or dried micro-bodies are formed into at least par of a free flowing powder. In a further specific embodiment the method, formulation and preservative according to the invention are characterized in that said active component has organoleptic properties that are masked by said at least one core inside said micro-body. The organoleptic properties may relate to either a taste or a smell. In a specific embodiment, said active component is a fragrance, perfume or a mixture of smelling compounds.

In another specific embodiment, said active component is a flavor or mixture of tasty compounds. In another specific embodiment said active component is a bad-tasting and/or bad-smelling compound of which the taste and/or smell is at least partially masked by the method, formulation, and/or preservative according to the invention.

In a further embodiment, a formulation comprising an active compound in a preserved state by means of the preservative according to the invention is characterized in that said formulation is a product or intermediate product for a cosmetic, a dermatological, food, nutritional, supplemental, agrochemical, fragrance, pharmaceutical, (bio)medical, household, energy storage, coating, or adhesives/glue application. More particularly, said formulation or said (intermediate) product is of detrimental nature to the pristine (i.e., unpreserved) active compound, more particularly characterized in that said formulation or said (intermediate) product comprises substantial amounts of water, more particularly >30 wt% water, more particularly >50 wt% water. More particularly, said formulation or said (intermediate) product is of detrimental nature to the pristine (i.e., unpreserved) active compound, more particularly characterized in that said formulation or said (intermediate) product has an acidic pH<7, more particularly pH<4. More particularly, the product is selected from the group of creams, lotions, serums, gels, shampoos, beverages, drinks, soda drinks, gummies, bars, tablets, capsules, pills, dairy, and injectables.

In a further embodiment, a formulation comprising an active compound in a preserved state by means of the preservative according to the invention is characterized in that said formulation is a product or intermediate product produced via a process that is damaging or detrimental to said active compound (especially when said active compound is not in said preserved state), more particularly wherein said process comprises one or more process steps performed at or above room temperature, more particularly at or above 40 degrees Celsius, more particularly at or above 60 degrees Celsius, more particularly at ore above 80 degrees Celsius, more particularly wherein one or more process steps are related to spray drying, hot melt, extrusion, injection molding, more particularly wherein the process involves one or more steps involving shear forces (e,g., mixing, blending, injection, extrusion, or centrifugation), and more particularly wherein the process is related to the production of (edible) gummies, also referred to as gummification.

The invention moreover relates to a cosmetic product, a personal care product or home care product, a dermatological product, a food or nutrition product, an agrochemical product, a fragrance product, a health or healthy product, a pharmaceutical product, a (bio)medical product, a household product, an energy storage product, a coatings product, and to an adhesives product that comprises the above formulation with an active compound in preserved form. Some of these will be described in further detail with reference to a number of embodiments:

Hereinafter, the invention will be described in further detail with reference to a specific embodiment and an accompanying drawing. In the drawing:

Figure 1 shows exemplary images of phase separation of an LAA-laden oleogel at different wax fractions;

Figure 2 gives a quantification of the phase separation shown in figure 1;

Figure 3 shows visual aspects of multiple capsules mixed with an aqueous base liquid;

Figure 4 shows a schematic representation of a micro-droplet encapsulated by a polymer shell according to the invention;

Figure 5 shows viable and multiplying probiotic cells expressed in colony-forming units (CFU) per 1 gram of the original excipient material after 1, 5, and 13 weeks, respectively, on stability tests in comparison to non-encapsulated probiotics in water and exposed to environmental air at 25 degrees Celsius (N.D. indicates not detectable);

Figure 6 shows a graph of a comparison of the stability of encapsulated probiotic L. Rhamnosus over time during accelerated stability tests in the following core formulations:

PB beads 1: 5% carnauba oleogel in safflower oil; probiotic powder dispersed directly into oleogel at 45°C;

PB beads 2: 5% carnauba oleogel in safflower oil; probiotic powder in oil mixed in a 1:1 ratio into oleogel at 50°C;

PB beads 3: 5% candelilla oleogel in safflower oil; probiotic powder dispersed directly into oleogel at 45°C; and

The viable and multiplying probiotic cells are expressed in colony-forming units (CFU) per 1 gram of the original excipient material after 13 weeks of stability tests. Encapsulated probiotics were incubated in water at 25°C and extracted from capsules at defined time points;

Figure 7 shows relative anti-oxidant activities of encapsulated versus unencapsulated active LAA under different stability test conditions in time.

It is noted that the figures are drawn purely schematically and not necessarily to a same scale. In particular, certain dimensions may have been exaggerated to a more or lesser extent to aid the clarity of any features. Similar parts are generally indicated by a same reference numeral throughout the figures.

Figure 4 shows a schematic representation of a preservative according to the invention. The preservative comprises one or more micro-bodies as shown in the figure that are encapsulated by a polymer shell according to the invention. The micro-body comprises a core 10,20 surrounded by a shell 30. The shell 30 is formed by a polymer network.

Said core 10,20 comprises an at least substantially solidified, particularly gelled, gellable or crystallizable liquid hydrophobic material 20, particularly an oleogel. A solid powder 10 is dispersed throughout said oleogel 20 and comprises said active compound. In principle the micro-body or micro-droplet according to the invention can be realised in a wide range of dimensions, ranging from sub-micron like nano proportions to a few millimetre. The application of the active compound has been found particularly favourable, if provided in the form of such micro or nano micro-bodies, having sub-micron to millimetre size. Accordingly said shell layer 30 may enclose a volume of between the order of one femtolitre and the order of one to several millilitres. Similarly the micro-droplet 10,20,30 has a diameter of the order of between half a micron and a centimetre, particularly between 5 micron and 5 millimetre, particularly between 100 and 500 micron, even more particularly between 150 and 350 micron, between 500 and 5000 micron, between 1 and 5 millimetre or between 2 and 4 millimetre.

The micro-bodies may be substantially spherical, having a sphericity of over 0.8, particularly beyond 0.9, and more particularly exceeding 0.95. The micro-body or microdroplet may, however, be not perfectly spherical, its diameter being represented by its average Feret diameter determined using, for instance, microscopic or macroscopic imaging in combination with dynamic or static image analysis. Alternatively, the diameter may be determined using laser diffraction, dynamic ultrasonic extinction, or dynamic light scattering.

When measured using a non-3D microscopic method, for example, when measured using standard bright field or phase-contrast microscopy, the sphericity of the micro-bodies may be described as circularity. In that case, the micro-bodies may be substantially circular, having a circularity of over 0.8, particularly beyond 0.9, and more particularly exceeding 0.95.

Sphericity of a particle can be calculated by (n^Al/3 (6 Vp)^A2/3) / Ap, wherein Vp is the particle's volume, Ap is the particle's surface area, and a perfect sphere is valued by a sphericity of exactly one. Circularity of a particle can be calculated by 4 n (area/perimeter^A2), wherein a perfect circle is valued by a circularity of one. Preferably the micro-bodies have a substantially identical size within a size distribution (dispersity) showing a relative size distribution characterized as coefficient of variation (i.e., standard deviation divided by average) lower than 10%, particularly lower than 7.5%, more particularly lower than 5%. Relative size distributions lower than 10% will be called 'mono-disperse¹.

The suspension 20 may contain said powder 10 at a concentration higher than 0.01% weight/weight (wt%), particularly between higher than 1 wt%, particularly higher than 10 wt%, particularly between 0.01 and 65%, particularly 0.1-50 wt%, particularly 0.5-5 wt%, particularly 5-40 wt%, particularly 5-15 wt%, particularly approximately 10 wt%. A moisture content of said suspension may be lower than 40%, particularly lower than 30%, particularly lower than 20%, particularly lower than 10%.

A moisture content of the suspension may be lower than 40%, particularly lower than 30%, particularly lower than 20%, particularly lower than 10%. The water activity (aw) of said oleogel may be lower than 0.7, particularly lower than 0.6, particularly lower than 0.5, particularly lower than 0.4, particularly lower than 0.3, particularly lower than 0.2, particularly lower than 0.1. The water activity (aw) is the partial vapour pressure of water in a solution divided by the standard state partial vapour pressure of water. The water activity (aw) of the oleogel may be lower than 0.7, particularly lower than 0.6, particularly lower than 0.5, particularly lower than 0.4, particularly lower than 0.3, particularly lower than 0.2, particularly lower than 0.1.

Example 1:

To prepare an oleogel, sunflower oil was heated to 100°C and carnauba wax flakes were added while stirring. Once the wax flakes were completely dissolved, the melted wax/oil mixture was left at 100°C for 10 min and after that cooled down to 40 °C. Ultra-fine L-ascorbic acid (LAA) powder was added to the (partly) liquid wax/oil mixture while stirring to create a suspension. The suspension was placed into an ultrasonic bath kept at 40 °C on sweep mode for 45 minutes to further promote homogeneous dispersion of the LAA powder. The homogenized LAA/wax/oil suspension was then gelled to form

LAA-laden oleogel by incubating the suspension at 4 °C for at least 1 hour.

The stability of the LAA-laden oleogel systems was determined by measuring the relative amount of phase separation after 28 days incubation at 40 °C and > 80% humidity. Carnauba wax concentrations of 2.5 wt% (A), 3.5 wt% (B), 4.5 wt% (C and D), and 5.5 wt% (E) in sunflower oil where compared. LAA concentrations where varied from 10 wt% to 40 wt% in oleogel. Figure 1 (representative images) and Figure 2 (quantification of phase separation) clearly show that suspensions B-E that comprise over 3 wt% wax are significant more stable after 28 days at 40 °C and > 80% humidity than suspension A having less than 3% wt% wax.

Example 2:

To prepare an oleogel, 340 g sunflower oil was heated to 100 to 110 °C, after which 20 g candelilla wax was added. The mixture was mixed until the wax melted and then kept at 100 to 110 °C for 10 to 15 minutes. The mixture was cooled down to 60 °C while stirring. Then 40 g LAA was slowly added to the wax/oil mixture while mixing at 2500 rpm using an overhead stirrer.

After adding LAA, the mixture was stirred for at least 5 minutes at 2500 and subsequently 15 minutes at 2000 rpm while the temperature was kept between 50 and 60 °C. The LAA/wax/oil mixture was then further homogenized using a rotor-stator mixer (ultra-turrax) at 16000 rpm for 5 min using mixer intervals of 1 to 2 min to prevent the temperature from increasing above 60 °C. Between homogenization intervals, the mixture was continuously mixed by gentle agitation to keep a homogeneous suspension. The LAA/wax/oil suspension was kept between 50 and 60 °C at al times and until further use (i.e., encapsulation). Capsules were produced with a dripping set-up using two syringe pumps and a coaxial nozzle, comprising an inner channel co-axially surrounded by an annular outer channel. A syringe, prewarmed to at least 50 °C, was filled with the LAA/wax/oil suspension and connected via heat-traced tubing (set to at least 50 °C) to the core annulus of the coaxial nozzle. Another syringe (also prewarmed to at least 50 °C) was filled with a solution containing 0.75 wt% sodium alginate (SA), 0.5 wt% agar, and 0.5 wt% carboxy methyl cellulose (CMC) in water and connected via heat-traced tubing (set to at least 50 °C) to the outer annulus of the coaxial nozzle.

The syringes were emptied using a syringes pump that created a steady stream of compound liquid exiting the coaxial nozzle and pinching off into compound core-shell miucro-bodies comprising a core of LAA/wax/oil suspension and a shell of SA/agar/CMC solution. The falling miucro-bodies (due to gravity) were collected in an ice-cooled and continuously stirred bath with cross-linker solution comprising 0.2 M calcium chloride and 20 v/v% ethanol. After at least 15 minutes incubation, the cross-linked capsules were transferred to cross-linker liquid with calcium chloride (without ethanol) and stored at 4 °C overnight, which allowed the oleogel cores to further gel.

Example 3:

An alternative capsule production method was explored, using similar core and shell liquids (i.e., 20 wt% LAA / 5.0 wt% carnauba wax in sunflower oil suspension as the core liquid, and 0.75% SA / 0.5 wt% agar solution as the shell liquid). Specifically, capsules were produced using 'in-air microfluidics¹, as described in WO 2017/167798, using a heat-traced co-axial first nozzle set to at least 50 °C and a simple (single opening) second nozzle. The core and shell liquids were jetted from the heated co-axial nozzle, while cross-linker solution containing 0.2 M calcium chloride + 20 v/v% ethanol, was jetted from the second nozzle.

By vibrating the coaxial nozzle using an external actuator device that was driven by a waveform generator generating a sinus wave with a frequency of 100 Hz in combination with a signal amplifier, the liquid core/shell compound jet (with active/oil/wax dispersion in the core and hydrophylic polymer solution in the shell) was broken into substantially identical miucro-bodies. Such monodisperse compound droplet train or jet stream may typically be composed of substantially equally sized miucro-bodies with typically a coefficient of variation in size or diameter, i.e. a standard deviation divided by average, of less than 10%.

The stream of compound core-shell miucro-bodies (which were approximately 50 to 60 °C) was coalesced with the liquid jet containing cross-linker that was kept at approximately 4 °C, which caused the (partial) solidification of the shell via (physically or chemically) cross-linking the polymers in the shell, thereby forming SA/agar/CMC capsules with a core containing dry LAA powder suspended in a gelling oil/wax mixture. The coalesced miucro-bodies and jets were collected in a bath with ice-cold cross-linker liquid comprising 20% ethanol and 0.2 M calcium chloride. Some capsules were post-treated with 1 wt% citric acid, which can cross-link the CMC, but can also act as plasticizer for the calcium-alginate polymer network, altogether increasing the mechanical stability and yield stress of the capsules. The resulting capsules were mixed with another aqueous phase and the product stability was analysed by evaluating the appearance and color of the samples at several time points during accelerated stability testing at 40 °C and 60 °C and >80% humidity.

Figure 3 shows the visual aspect of multiple capsules mixed with an aqueous base liquid after 28 days of accelerated stability test (40 °C and >80% humidity). The left sample (Figure 3A,3B) contains capsules that have a shell comprising calcium-cross-linked sodium alginate and cores comprising 10 wt% dry LAA powder dispersed in oleogel consisting of 5 wt% carnauba wax in sunflower oil. The right sample (Figure 3C,3D) contains capsules that have a shell comprising calcium-cross-linked sodium alginate similar to the capsules in the left sample, and a core comprising carnauba-wax stabilized W/O emulsion including 10% LAA dissolved in the water phase. These capsules (Figure 3C,3D) have been prepared according to the method and design as described in International patent application WO2022/029623 by applicant.

From the images it can be clearly observed that both the capsules and the aqueous buffer surrounding the capsules in the left sample (Figure 3A,3B), which has been prepared according to the current invention are significantly less coloured than the capsules and aqueous buffer surrounding the capsules in the right sample (Figure 3C,3D). Since LAA degradation results in colouring, these data indicate that the method and preserving micro-body according to the invention give a better preservation of the active ingredient as compared to the method and preserving micro-capsules described in WO2022/029623.

Example 4:

Capsules were prepared similarly to the method in example 3, but using different core and shell liquids. Instead, the core composition contained 1 wt% probiotics (spray dried powder comprising mixed strains) suspended in a mixture of 4 wt% candelilla wax in sunflower oil, instead of LAA in carnauba wax / sunflower oil mixture.

Example 5:

Capsules were prepared similarly to the method in example 3, but using different core and shell liquids. The core composition contained 2 wt% non-spore forming probiotics (freeze dried Lactobacillus rhamnosus) suspended in a mixture of 5 wt% carnauba wax in safflower oil, instead of LAA in carnauba wax / sunflower oil mixture, and the shell comprising 1.5wt% SA, instead of SA/agar mixture.

After production, probiotic-laden capsules were subjected to stability tests, during which the capsules were incubated in calcium-containing aqueous medium at 25°C. At predefined time points (after 1, 5, 13 weeks), probiotics were extracted from the capsules using shear homogenization, inoculated on agar plates, and incubated for 48 hours at 37°C. The number of viable and multiplying probiotic cells was assessed through colony-forming unit (CFU) enumeration.

The samples were prepared in duplicate, and inoculation was performed in technical duplicate. Controls included non-encapsulated probiotics powder that was incubated in calcium-containing aqueous medium and incubation of the probiotics powder in environmental air with relative humidity of approximately 40 to 50%. Viability analyses (i.e., CFU counts) indicated that encapsulating probiotics in the capsules according to the invention significantly improved probiotic survival compared to non-encapsulated counterparts under stability testing conditions over time (Figure 5).

After 5 weeks of incubation, probiotics in water were observed to be non-viable (i.e., no detectable CFUs), and by week 13, all probiotics exposed to air had perished, while encapsulated probiotics remained viable. These findings suggest that the capsule formulation according to the invention offers protective properties against, amongst others, water, moisture and non-chilled (i.e., heat) conditions, which are indeed critical factors known to compromise probiotic viability.

Example 6:

Bi-gel capsules were prepared similarly to the method in example 3, but using different core and shell liquids. The core composition contained 2 wt% non-spore forming probiotics (freeze dried Lactobacillus rhamnosus) suspended in a mixture of 5 wt% carnauba wax in safflower oil, instead of LAA in carnauba wax / sunflower oil mixture, and the shell comprising 1.5wt% SA, instead of SA/agar mixture.

After production, the capsules where successfully dried by using a fluidized bed drier set at 35°C air inflow temperature (i.e., below melting point of oleogel) in the presence of 2 wt% aluminium silicate (used as anticaking agent) until a free-flowing powder was obtained. Viability analysis of extracted probiotics in combination with CFU counting revealed that "90% of probiotics survived the drying process.

Example 7:

Capsules were prepared similarly to the method in example 3, but using different core and shell liquids. The core composition contains 2 wt% probiotics (freeze dried L. rhamnosus) suspended in a mixture of 5 wt% in carnauba wax in safflower oil instead of LAA in carnauba wax / sunflower oil mixture, and the shell comprising 1.5wt% SA, instead of SA/agar mixture, shown as "PB beads 1" in figure 6. -Tl-

In this example, the wax/oil mixture was first heated to 90-100 degrees Celsius to form a transparent and homogeneous liquid, and then cooled down to 45 degrees Celsius before probiotics containing powder was added and mixed through. Furthermore, capsules with the same composition as PB beads 1 were made using a different core liquid preparation method that resulted in a less homogeneous distribution of wax throughout the oleogel core, shown as "PB beads 2" in figure 6.

Specifically, for PB beads 2, a 10% carnauba wax/safflower oil mixture without probiotics was heated to 90-100 degrees Celsius and then cooled to 50 degrees Celsius, after which probiotics-laden oil at room temperature (i.e., "20-25 degrees Celsius) was added in a 1:1 ratio to reach a final carnauba concentration of 5%. Blending the warmer wax/oil mixture with the colder probiotics-laden oil presumably resulted in substantial wax crystallization and oleogelation prior to the in-air microfluidics encapsulation process, and eventually resulting in a less homogeneous and less protective oleogel network surround the probiotics.

Furthermore, another batch of probiotics-laden capsules was prepared in a similar manner as PB beads 1, but then the core composition contained probiotics in a mixture of 4 wt% candelilla wax in safflower oil, shown as "PB beads 3" in figure 6.

Analysing probiotics viability after stability testing the capsules in an aqueous base at 25 degrees Celsius revealed that probiotics survival in PB beads 1 was significantly higher than in PB beads 2 and PB beads 3, indicating that both the formulation and encapsulation process parameters play an important role in the final degree of protection against and prevention of probiotics death, as well as extension of shelf life, particularly in a wet or moist application.

Example 8:

Microcapsules were prepared similarly to the method in example 3, but using different core and shell liquids. Instead, the core composition comprised 10wt% LAA and 5wt% Carnauba wax in high-oleic sunflower oil and the shell composition comprised (gelled) SA/agar/CMC. The capsules were provided as a 50-60% slurry in an aqueous medium in a wide-neck high-density polyethylene container and subsequently subjected to and passed a transportation test according to ASTM D4169-16 before being put on stability tests.

For stability testing, the capsules were retrieved from the container, separated from the aqueous transport medium by sieving, blended into a various water-based liquids and gels, and subjected to different storage conditions at 4, 21, 30, 40, and 60 ± 2 degrees Celsius for a minimum of 3 months. LAA was extracted from the capsules using ultrasonic treatment in ethanol and anti-oxidant activity of the extracted LAA was measured using the 2,2-diphenyl-l-picrylhydrazyl assay in combination with spectrophotometry.

The encapsulated active LAA shows significantly higher anti-oxidant activity and slower reduction of anti-oxidant activity, as well as longer anti-oxidant half-life time of anti-oxidant activity as compared to unencapsulated active LAA (i.e., control) (Figure 7).

Although the invention was explained in more detail above on the basis of merely a limited number of examples and embodiments, it should be clear that the invention is by no means limited to that. On the contrary, many variations and embodiments are still possible within the scope of the invention for an average person skilled in the art.

As an example, the method, formulation and preservative according to the invention may be used in embodiments, wherein said active compound comprises at least one vitamin, particularly a vitamin that is selected from a group containing thiamine, riboflavin, nicotinic acid, pantothenic acid, pyridoxine, biotin, folic acid, cyanocobalamin, lipoic acid, ascorbic acid, lecithin, glycyrrhizin acid, retinol, retinol palmitate, tocopherol, tocopherol acetate, salicylic acid, benzoyl peroxide, and azelaic acid and/or derivatives thereof, more particularly ascorbic acid and/or derivatives thereof. Also the method, formulation and preservative according to the invention may be used in embodiments, wherein said active compound comprises at least one anti-oxidant, particularly an anti-oxidant that is selected from a group containing poly-phenols, thiol-based components, sulphite and derivatives thereof.

The active compound may be used, for instance, as nutritious supplement or for pharmaceutical treatment, in which case it is likely to be administered orally. The shell layer may be formulated to survive the acidic environment of the human stomach to be digested in the more downstream portion of gastrointestinal tract of the user to release its contents. Particularly, the shell layer may comprise a digestible or fermentable polymer, more particularly the shell layer may comprise a pectin compound. Specifically pro- and prebiotics may be administered particularly effectively in this manner.

Particularly, said shell layer may carry a coating, preferably an edible coating, particularly a hydrophobic coating containing nano-particles or a wax like carnauba wax.

Claims

Claims:

1. Method for preservation of an active compound, wherein a powder comprising said active compound is provided, wherein said powder is dispersed in an at least partly gellable or crystallizable liquid hydrophobic material to form a suspension or dispersion containing said powder, wherein said suspension or dispersion is formed into micro-bodies, wherein said micro-bodies are encapsulated by a polymer shell to form suspension- or dispersion-laden micro-bodies which contain one or multiple cores of said suspension or dispersion, wherein said suspension- or dispersion-laden micro-bodies are at leat partly solidified, particularly gelled and/or crystallized, to form micro-bodies containing said active compound in an at least substantially solidified hydrophobic network.

2. Method according to claim 1, characterized in that said liquid hydrophobic material is processed in a liquid condition to at least partly solidify or have solidified below a solidification temperature thereof.

3. Method according to claim 2, characterized in that said liquid hydrophobic material is cooled down to below said solidification temperature thereof at a cooling rate faster than 0.1 K/min, particularly faster than 1 K/min, more particularly faster than 10 K/min, and more particularly faster than 100 K/min.

4. Method according to claim 2 or 3, characterized in that said liquid hydrophobic material is solidified by combining the micro-bodies with a phase, particularly a liquid phase, that has a temperature lower than a solidification temperature of said liquid hydrophobic material, particularly a temperature below room temperature, more particularly a temperature below 10 degrees Celsius.

5. Method according to claim 4, characterized in that said phase comprises a liquid phase containing a cross-linking compound to cross-link said shell polymer.

6. Method according to claim 5, characterized in that said liquid phase has a temperature that induces thermal or physical cross-linking of at least on of said shell polymers.

7. Method according to anyone of the preceding claims, characterized in that said one or more micro-bodies are laden with a hydrophobic compound by immersing said one or more micro-bodies into a solution of said hydrophobic compound, particularly a concentrated solution, allowing said hydrophobic compound to diffuse trough said shell into said one or more cores of said one or more micro-bodies.

8. Method according to anyone of the preceding claims, characterized in said one or more micro-bodies are laden with a hydrophobic compound by providing said one or more micro-bodies with one or more cores in molten state prior to immersing said one or more micro-bodies into a solution of said hydrophobic compound, particularly a concentrated solution, to allow said hydrophobic compound to diffuse into said one or more cores of said one or more micro-bodies in said molten state, and subsequently allowing said one or more cores to at least substantially solidify.

9. Method according to anyone of the preceding claims, characterized in that said micro-bodies are dried, particularly by a drying method chosen from a group containing evaporation, blowing dry gas, vacuum drying, fluid bed drying, freeze drying, microwave drying, and chemical drying.

10. Preservative for an active compound, comprising one or more preserving micro-bodies, each having at least one core surrounded by a shell that is formed by a polymer network, wherein said core comprises an at least substantially solidified, particularly gelled, gellable or crystallizable liquid hydrophobic material, particularly an oleogel, and a solid powder, said powder being dispersed throughout said core, and wherein said powder comprises said active compound.

11. Formulation containing an active compound in a preserved state by means of the preservative according to claim 10.

12. Formulation according to claim 11, being applied in or as a cosmetic product, a personal care or home care product, a dermatological product, a food or nutrition product, an agrochemical product, a fragrance product, a health or healthy product, a pharmaceutical product, a (bio)medical product, a household product, an energy storage product, a coatings product, or an adhesives product.

13. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said shell comprises a calcium-alginate network that is weakened or completely removed prior to or during final application, particularly by scavenging calcium ions and at least partly disrupting the calcium-alginate network of the shell, more particularly by introducing a chelator agent such as citric acid or ethylene-diamine-tetra-acetic acid (EDTA).

14. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said shell is designed to disintegrate or break under induced chemical or mechanical conditions, upon which said active compound comprising powder is released from the micro-body.

15. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said shell comprises a polymer network, particularly an interpenetrating network, double network, and/or composite of two or more inter- or intra- cross-linked polymers, and more particularly in that said polymer network comprises a hydrophilic polymer network, particularly comprising one or more poly-electrolytes or polysaccharides selected from agar, alginate, chitosan, dextran, poly(ethylene glycol), collagen, gelatin, hyaluronic acid, carrageenan, particularly Lambda-, Kappa and Lota carrageenan, fibroin, fibronectin, poly-l-lysine (PLL), cellulose, graphene, poly(ethylenimine) (PEI), poly(amidoamine) (PAA), dextran sulfate, silk, silk fibroin, pectin, locust bean gum, gellan gum, guar gum, tragacanth gum, xanthan gum, acacia gum, karaya gum, starch, and sodium carboxymethyl cellulose (S-CMC), all of these as naturally derived materials and/or synthetically derived materials including recombinant proteins and/or derivatives of these materials.

16. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said shell comprises a bio-compatible, bio-degradable and/or bio-resorbable polymer network, particularly a polymer network comprising a methyl-methacrylate derivative, a caprolactone derivative, a lactic acid derivative, a glycolic acid derivative, and/or a co-polymer of lactic acid and glycolic acid, more particularly in that said polymer network comprises a poly( lactic-co-glycol ic acid), a poly(caprolactone) and/or a poly(methyl-methacrylate).

17. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said polymer network comprises a cross-linked or inter-penetrating alginate network, particularly a calcium cross-linked alginate network.

18. Method, preservative or formulation according to claim 17, characterized in that network is strengthened by incorporation of nano-particles, micro-particles and/or poly-electrolytes.

19. Method, preservative or formulation according to claim 18, characterized in that said polymer shell comprises an alginate network, particularly a sodium-, calcium- and/or a shellac reinforced alginate network.

20. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material comprises at least one oil, particularly one or more oils from a group comprising: vegetable oils, such as sunflower oil, corn oil, linseed oil, canola oil, castor oil, palm oil, coconut oil, avocado oil, sweet almond oil, calophylum oil, sesame oil, olive oil, jojoba oil, soybean oil, cottonseed oil, rapeseed oil, peanut oil, flaxseed oil, borage oil, safflower oil, macroalgae oil and seaweed oil; essential oils, ethereal oils, macerated oils, triglyceride, animal oils, such as tallow fat, lanolin and marine oils, such as fish oils; synthetic oils, and neutral oils like medium chain triglyceride oils, as well as mixtures or derivatives thereof, and more particularly oils that are non-toxic and user-friendly, particularly sunflower oil or safflower oil.

21. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said hydrophobic material is an oleogel comprising an oil, more particularly comprising sunflower oil, that retains more than 90% of said oil, particularly more than 95% of said oil, during at least four weeks at 40 degrees Celsius.

22. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material comprises a hydrophobic gelling agent, specifically a fatty acid, a wax, or a sterol that is provided in liquid form and that is gelled or solidified into said micro-bodies, particularly one or more fatty acids and/or waxes from a group comprising: paraffin wax, rice bran wax, sunflower wax, carnauba wax, candelilla wax, beeswax, microcrystalline wax, coconut wax, ozocerite wax, beta-sitosterol, gamma-oryzanol, stearic acid, palmitic acid, behenic acid, myristic acid, lauric acid, capric acid, and fatty acid derivatives, such as fatty acid esters with short chain alcohols, such as isopropyl myristate, isopropyl palmitate and isopropyl stearate and dibutyl adipate, and particularly carnuba wax.

23. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material comprises a hydrophilic gelling agent, in particular a polysaccharide, a (modified) starch, a protein, a natural gum, a hydrocolloid, a steroid, a hydrophilic phytosterol derivative, or a phospholipid, in particular one or more gelling agents selected from the group of cellulose, maltodextrin, dextran, hyaluronic acid, gelatin, whey protein, gum Arabic, gum tragacanth, xanthan gum, carrageenan, agar, lecithin, propylene glycol, silica (nanoparticles), more particular ethyl cellulose.

24. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material has a solidification temperature that is between 0 and 100 degrees Celsius, particularly between 4 and 90 degrees Celsius, particularly between 4 and 40 degrees Celsius, particularly between 20 and 40 degrees Celsius, particularly between 20 and 90 degrees Celsius, particularly between 40 and 90 degrees Celsius, particularly between 40 and 50 degrees Celsius, particularly between 50 and 60 degrees Celsius, particularly between 60 and 70 degrees Celsius, particularly between 80 and 90 degrees Celsius, and preferably above room temperature.

25. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material has a solidification offset temperature below about 60 degrees Celsius.

26. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material comprises an emulsifier or plasticizer, such as fatty acids, glycolipids, monoglycerides, diglycerides, triglycerides, or phospholipids, or a mixture thereof, particularly soy lecithin or polyglycerol polyricinoleate (PGPR). 1. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material comprises an oil-soluble anti-oxidizing agent, such as, for example, a tocopherol (known as vitamin E), particularly alpha-tocopherol, particularly 0.01-10 wt% alpha-tocopherol in oleogel, particularly 0.1-1% alpha-tocopherol in oleogel.

28. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder is provided as a dry, substantially water-free powder.

29. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises an anti-oxidizing agent and/or a nutrient, particularly one or more from the group of catalase, polyphenol, curcumin, quercetin, catechin, lignan, resveratrol, citric acid, and L-ascorbic acid. In particular said powder comprises L-ascorbic acid.

30. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said active compound is a hydrating agent, particularly hyaluronic acid, alginate, gelatin or collagen.

31. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises a hydrophilic polymer with a molecular weight larger than 10 kDa, particularly larger than 100 kDa, particularly larger than 1000 kDa, in particular a polysaccharide, in particular hyaluronic acid.

32. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises a hygroscopic compound, particularly a natural compound, particularly from the group of hydrophilic polymers, polysaccharides, proteins, nucleic acids, and water-soluble salts.

33. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises a hygroscopic salt, in particular a salt of a quaternary ammonium cation, more particularly a choline salt, in particular one from the group of choline chloride, choline bitartrate, choline borate, choline dihydrogen citrate, choline bicarbonate and choline chloride carbamate, even more particularly choline chloride.

34. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises a biological compound, particularly an organism, a protein, an enzyme, a peptide, a nucleotide, or a mixture thereof.

35. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises a viable compound, particularly a living cell, more particularly a bacteria or microbiota.

36. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises one or more bacteria from the group of Bacteroides, Bifidobacterium, Fusobacterium, Bacillus, Lactobacillus, Saccharomyces, Streptococcus, Enterococcus Porphyromonas, Prevotella, Actinomyces, Propionibacterium, Clostridia, particularly one or more from the group of Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Saccharomyces boulardii, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus fermentum, Bacillus coagulans, Bacillus subtilis, Enterococcus faecium.

37. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises one ore more prebiotics, particularly one ore more from the group of oligosaccharides, fructans, galactans, starch, pectins, and beta-glucans.

38. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises a catalyst, in particular one or more compound from a group of compounds: that catalyse amino cross-linking, particularly comprising p-TSA (para-Toluene Sulfonic Acid), DNNDSA (Di Nonyl Naphtalene Di Sulfonic Acid), DDBSA (Do Decyl Benzene Sulfonic Acid), DNNSA (Di Nonyl Naphtalene mono Sulfonic Acid), and Phosphate Acid, and Carboxylic Acid; or that catalyse urethane cross-linking, particularly including DBTL (Di Butyl Tin Laurate), DOTL (Di Octyl Tin Laurate), DBTO (Di Butyl Tin Oxide), Bismuth-based catalyst, Zirconium-based catalyst, and amine-based catalyst. 39. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said liquid hydrophobic material is an oleogel comprising an oil, more particularly comprising an oil with an oleic acid content higher than 10%, more particularly higher than 25%, more particularly higher than 50%, more particularly higher than 70%, even more particularly comprising high-oleic sunflower oil characterized by an oleic acid content higher than 70%.

40. Method, preservative or formulation according to claim 39, characterized in that said oleogel retains substantial amounts of said oil, more particularly wherein said oleogel has the capacity to bind or retain more than 90% of said oil in said oleogel, particularly more than 95% of said oil in said oleogel, during at least 4 weeks at or above room temperature, particularly at 40 degrees Celsius.

41. Method, preservative or formulation according to claim 39 or 40, characterized in that said oleogel comprises an oil gelling agent at a concentration of at least 1 wt% (gelling agent to oleogel weight percentage), particularly at least 2 wt%, particularly at least 3 wt%, particularly at least 4 wt%, particularly at least 5 wt%, particularly between 1 wt% and 10 wt%, particularly between 4 wt% and 6 wt%.

42. Method, preservative or formulation according to claim 39, 40 or 41, characterized in that said a water activity (aw) of said oleogel is lower than 0.7, particularly lower than 0.6, particularly lower than 0.5, particularly lower than 0.4, particularly lower than 0.3, particularly lower than 0.2, particularly lower than 0.1.

43. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said micro-bodies comprise micro-droplets or nano-droplets, having sub-micron to millimetre size.

44. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said shell encloses a volume of between the order of one femtolitre and the order of one to several millilitres.

45. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said micro-bodies have a diameter of the order of between half a micron and a centimetre, particularly between 5 micron and 5 millimetre, particularly between 100 and 500 micron, even more particularly between 150 and 350 micron, between 500 and 5000 micron, between 1 and 5 millimetre or between 2 and 4 millimetre.

46. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said micro-bodies are substantially spherical, having a sphericity of over 0.8, particularly beyond 0.9, and more particularly exceeding 0.95.

47. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said micro-bodies have a substantially identical size within a size distribution (dispersity) showing a relative size distribution characterized as coefficient of variation lower than 10%, particularly lower than 7.5%, more particularly lower than 5%.

48. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said suspension contains said powder at a concentration higher than 0.01% weight/weight (wt%), particularly between higher than 1 wt%, particularly higher than 10 wt%, particularly between 0.01 and 65%, particularly 0.1-50 wt%, particularly 0.5-5 wt%, particularly 5-40 wt%, particularly 5-15 wt%, particularly approximately 10 wt%.

49. Method, preservative or formulation according to anyone of the preceding claims, characterized in that a moisture content of said suspension is lower than 40%, particularly lower than 30%, particularly lower than 20%, particularly lower than 10%.

50. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said a water activity (aw) of said suspension is lower than 0.7, particularly lower than 0.6, particularly lower than 0.5, particularly lower than 0.4, particularly lower than 0.3, particularly lower than 0.2, particularly lower than 0.1.

51. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises a dried, spray dried, or freeze dried composition comprising an active ingredient (hygroscopic) drying excipient, particularly a drying excipient comprising at least one polycarbohydrate, monosaccharide, disaccharide, or polysaccharide compound, and more particularly a drying excipient being a sugar, more particularly the drying excipient being selected from the group of dextran, dextrin, maltodextrin, trehalose, lactose, glucose, dextrose, sucrose, fructose, maltose, isomaltose, sorbitol, mannitol, lactitol, xylitol, and/or erythritol.

52. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises one or more sensitive bacteria selected from the group of anaerobic bacteria, non-spore forming bacteria, and other bacteria sensitive to moisture, acid, heat, and/or oxygen.

53. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said powder comprises one or so called 'next-generation probiotics', more particularly said powder comprises one or more next-generation probiotics from the group of Akkermansia muciniphila, Faecalibacterium prausnitzii, Bacteroides thetaiotaomicron, Bacteroides fragilis, Roseburia spp., Prevotella spp., Alistipes spp., Christensenella minuta, Blautia spp., Eubacterium hallii, and Methanobrevibacter smithii

54. Method, preservative or formulation according to anyone of the preceding claims, characterized in that said active component has organoleptic properties that are masked by said at least one core inside said micro-body.

55. Method, preservative or formulation according to claim 54, characterized in that said active component is a fragrance, a perfume or a mixture of smelling compounds.

56. Method, preservative or formulation according to claim 54, characterized in that said active component is a flavour or comprises a mixture of tasty and bad-tasting compounds.