WO2020237383A1 - Thylakoids as delivery system for cannabinoïds and other molecules and formulations thereof - Google Patents

Abstract

There is provided the use of extracted active/functional thylakoid membranes from photosynthetic organisms, particularly as vesicles for the formulation of liposoluble compounds such as, for example, THC and other cannabinoids extracted from cannabis. There is further provided a composition in pharmaceutical application, useful for the treatment or prevention of various conditions that may be addressed by cannabinoids (Cbs), made bioavailable or potentiated via formulation in thylakoid membranes. Method for the production thereof are also provided.

Description

THYLAKOIDS AS DELIVERY SYSTEM FOR CANNABINOIDS AND OTHER MOLECULES AND FORMULATIONS THEREOF

Field

[0001] The present invention relates to thylakoids and functional derivatives, as carriers for molecules. More specifically, this invention relates to a composition comprising functional thylakoids, particularly in specific formulations that ensure the integrity and functionality of molecules, particularly liposoluble molecules that are difficult to formulate, to increase their bioavailability. Still, more particularly, the invention concerns a method for the simultaneous extraction of cannabinoids (CBs) embedded in thylakoid membranes.

Background

[0002] The regulation of cell-signaling and physiological mechanisms is governed by several biochemical reactions depending on changes occurring in biological membranes. Indeed, biological membranes, consisting essentially of phospholipid bilayers ensuring their fluidity, contain globular proteins and glycoproteins that are intercalated among lipids (Nicolson, 2014). The biological membranes therefore represent a confinement barrier in which membrane proteins are ligands binding specifically to their molecular targets. Thus, the simultaneous interactions between the lipid bilayers and the extracellular media, which are provided by membrane proteins, allow the regulation of physiological and biochemical mechanisms. (Dainese et a!., 2010).

[0003] Thylakoids are membrane-bound compartments inside chloroplasts and cyanobacteria. They are the site of light-dependent reactions of photosynthesis. In higher plants and algae, thylakoid membranes form a network of aggregated membranes with semi- cylindrical structures, from 5 to 20 disks called grana. They are interrelated by non-stacked membrane regions, the lamellae (Pfeil et al. 2014). The outer surface of the thylakoids is immersed in the stroma while the internal side surrounds an aqueous compartment, the lumen.

[0004] Thylakoid membranes contain the photosynthetic apparatus of chloroplasts and constitute the organelles responsible of photosynthesis. They consist of important protein complexes comprising the photosystems I and II protein complexes (PS I and PS II), cytochrome b6/f complex and the complex of ATP synthase. The particular organization of thylakoids membranes promotes the optimal activity of the two photosystems I and II lipoprotein complexes, in which photochemical reactions of photosynthesis take place.

[0005] Like all other biological membranes, thylakoid interactions are not limited to their specific targets. Other molecules such as pollutants can also interfere with these membranes. The organization of thylakoids allows toxic molecules to bind to any sites on the protein complexes thereby inhibiting photosynthetic electron transport. Herbicides that activate photosystem II bind to the site of a plastoquinone (QB) preventing electrons to move toward the cytochrome b6/f complex, thereby disabling the light reactions of photosynthesis (Boger and Sandmann, 2000; Draber et ai 1991 ; Kleczkowski, 1994; Prevot et ai, 1993; Wilkinson et ai, 2015). These herbicides are known under the name of atrazine, diuron, fluometuron, tebuthiuron, ametrynm metribuzin, simazinem, prometryn, bromacil, hexazinone. Another herbicide, paraquat, targets PSI and cytochrome b6/f complexes (Fujii et ai., 1990).

[0006] It is possible to prepare thylakoid membranes containing the entirety of the photosynthetic system in its active/functional form. Thylakoids have been used in a number of applications that take advantage of their ability to generate a photosynthetic response to outside factors (i.e. to scavenge reactive oxygen species), or to act as biosensors. WO 01/49305 discloses anti-oxidative compositions and method for their extraction. WO 03/04042 discloses their use in combination with other anti-inflammatory compounds.

[0007] By conventional soft methods such as mechanical breakage of plant cells and chloroplasts, it is possible to extract these membranes and to form lipid vesicles that maintain the physiological properties of the photosystems. Therefore, in addition to living cells, algae, and cyanobacteria, active / functional thylakoids membranes prepared by these methods provide excellent photosynthetic materials and provide interesting additional functional properties.

[0008] Due to its lipid matrix and its ability to interact with insoluble molecules such as herbicides and PAH, galactolipid-forming thylakoid membranes provide an excellent candidate matrix for inserting and concentrating other lipophilic molecules such as, for example, cannabinoids (Cb). Indeed, plant cannabinoids are practically insoluble in water but soluble in lipids, alcohols, and other non-polar organic solvents.

[0009] It is known that Cb interact with biological membranes by amphipathic bonds with membrane lipids such as phospholipids (Yang et ai, 1992) containing both saturated and unsaturated fatty acids (Bruggemann and Melchior, 1983). Hemp seed oil has also been formulated in emulsions composed of different fatty acids (Mikulcova et ai, 2017).

[0010] Therefore, here is an opportunity to further exploit the properties of thylakoids in novel applications, such as for example as carriers for the making of new formulations of useful lipophilic compounds, such as Cb.

SUMMARY

[0011] Therefore, in accordance with the present invention, there is provided a composition comprising thylakoid vesicles stably incorporating a lipophilic compound.

[0012] Therefore, in accordance with the present invention, there is provided a composition comprising a lipophilic compound embedded in thylakoid vesicles.

[0013] Still, more particularly, the thylakoids are extracted from the same plant in which the lipophilic molecule is extracted in such a way that the lipophilic molecule is embedded in the thylakoid during a single extraction.

[0014] In a particular embodiment, the lipophilic molecule is extracted from a Cb- producing plant, such as Cannabis sativa, indica or hemp. In a particular embodiment, the thylakoid membranes/vesicles are extracted from a Cb-producing plant, such as Cannabis sativa, indica or hemp. Still, more particularly, the Cb and the thylakoid membranes/vesicles are extracted from the same Cb-producing plant, such as Cannabis sativa, indica or hemp.

[0015] More particularly, a Cb-containing thylakoid extract is provided. More particularly, there is also provided a Cb-containing thylakoid extract, wherein the thylakoids have the capacity to be activated in presence of electron donors.

[0016] Particularly, there is provided a stabilized Cb-containing thylakoid extract.

[0017] The stabilized thylakoid extract may be dried or in a medium composed of an amphoteric solvent such as propylene glycol or PVP.

[0018] In accordance with another aspect of the present invention, there is provided a pharmaceutical composition comprising the Cb-containing thylakoid extract of the present invention, in admixture with a pharmacologically acceptable carrier. [0019] In accordance with another aspect of the present invention, there is provided thylakoids or thylakoid functional derivatives for use as carriers for lipophilic compounds, such as, for example, cannabinoids.

[0020] In accordance with another aspect of the present invention, there is provided a use of thylakoids or thylakoid functional derivatives as carriers for lipophilic compounds, such as, for example, cannabinoids.

[0021] According to a particular aspect, this invention provides a method for producing a Cb-comprising thylakoid extract comprising the steps of combining a CB extract with a thylakoid suspension.

[0022] According to a particular aspect, this invention provides a method for producing a Cb-comprising thylakoid extract comprising the steps of extracting thylakoids from a Cb producing plant part, under conditions to recover Cb in said thylakoid extract.

[0023] Other objects, advantages and features of the present invention will become more apparent upon reading of the following non-restrictive description of preferred embodiments thereof, given by way of example only with reference to the accompanying drawings.

[0024] The present description refers to a number of documents, the content of which is herein incorporated by reference in their entirety.

Detailed description

Abbreviations and Definitions

Abbreviations

[0025] Cb(s): cannabinoid(s); THC: tetrahydrocannabinol.

Definitions

[0026] The term“about” as used herein refers to a margin of + or - 10% of the number indicated. For the sake of precision, the term about when used in conjunction with, for example: 90% means 90% +/- 9% i.e. from 81 % to 99%. More precisely, the term about refer to + or - 5% of the number indicated, where for example: 90% means 90% +/- 4.5% i.e. from 86.5% to 94.5%. [0027] As used herein, the term "a" or "the" means "at least one".

[0028] As used herein the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and reference to "the culture" includes reference to one or more cultures and equivalents thereof known to those skilled in the art, and so forth. All technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this invention belongs unless clearly indicated otherwise.

[0029] The term“Cannabis” as used herein includes Cannabis Sativa, Cannabis Indica (drug-producing cannabis) and includes hemp also called industrial hemp or fiber-producing cannabis in contrast to drug-producing cannabis. It is generally recognized that strains from Cannabis Sativa and Cannabis Indica produce cannabinoids such as: tetrahydrocannabinol (THC) and THC-derived compounds and cannabidiol (CB) and CB-derived compounds such as presented in Figures 1 and 2. On the other hand, hemp is known to produce the same variety of CBs, but without a significant production of THC (less than 0.3%).

[0030] The term“Cannabinoids” (Cbs) as used herein includes tetrahydrocannabinol and all other cannabinoids, all in their acid and in their decarboxylated forms. For example, THC (A9-tetrahydrocannabinol), CBD (Cannabidiol), CBG (Cannabigerolic), CBC (Cannabichromenene), CBGV (Cannabigerovarine), THCV (Tetrahydrocanabivarine), CBV (Cannabidivarine), CBCV (Cannabichromevarine); and all other compounds listed in Figure 3. As used herein, the abbreviation Cbs also includes THC.

[0031] As used in this specification and claim(s), the words“comprising” (and any form of comprising, such as“comprise” and“comprises”),“having” (and any form of having, such as“have” and“has”),“including” (and any form of including, such as“includes” and“include”) or“containing” (and any form of containing, such as“contains” and“contain”) are inclusive or open-ended and do not exclude additional, un-recited elements or method steps.

[0032] The terms“cosmetic” or“cosmeceutic” means an activity for the maintenance of external health, such as, for example, skin, hair or nails, whether it is administered externally or topically (e.g. by way of cream, lotion, shampoo or serum, etc.) or internally by way of nutritional supplement (cosmeceutic). [0033] The expression“photosynthetic organism” as used herein means organism that are able to convert light energy into chemical energy that can later be released to fuel the organisms' activities. Most plants, most algae, and cyanobacteria perform photosynthesis, including phytoplankton. Particularly, plant parts that are able to convert light to energy can be used to extract thylakoid therefrom, such as, for example: lead shoots, stem from vegetative plant, de-leafed flowers, full-dressed flowers, full plant from flowering plant; leaves from flowering plant; root from flowering plant; and/or stem from flowering plant. Alternatively, plant parts that contain or produce trichomes can also be used in the process of the present invention to produce CB-containing extract or solution, to be later added to the thylakoids during their extraction process. Trichomes are actually the very factories that produce the hundreds of known cannabinoids, terpenes, and flavonoids that provide cannabis strains their unique profiles. For example, trichomes-containing cannabis parts mostly from leaves and flowers (or buds).

[0034] The terms: ’’thylakoid”, “thylakoid extract”, “functional thylakoid extract” or “functional thylakoid” or“active thylakoids / extract” as used herein, means purified functional photosynthetic pigments in a thylakoid membrane environment (i.e. in an integral native state such that they can still be active or activated), particularly their original thylakoid environment. More particularly, these terms refer to functional thylakoid membranes as extracted by the process herein described and/or by the procedure disclosed in Bissonnette et al. (2004) or W001/49305. Particularly, in connection with an aspect of the present thylakoid extract, the functional quality of the molecular complex can be measured by fluorescence based on its capacity to react to light and dissipate its energy (Fv/Fm ratio), as is well known in the art and/or described in Maxwell (2000).

[0035] As such, a "therapeutically effective" or "prophylactically effective" amount of an agent capable of reducing the tumor, or a combination of such agents, may be administered to an animal, in the context of the methods of treatment and prevention, respectively, described herein.

[0036] A "subject in need thereof” or a "patient" in the context of the present invention is intended to include any subject that will benefit or that is likely to benefit from PDT or more generally from the thylakoids of the invention stably incorporating heterogeneous agents. In an embodiment, a subject in need thereof is a subject diagnosed with a cancer. Description of the figures

[0037] Figure 1 represents the metabolic pathways of THC and Cb production in the plant.

[0038] Figure 2 Biosynthetic pathway resulting in production of cannabinoid acids. Each cannabinoid acid can be subsequently decarboxylated into a corresponding cannabinoid, such as THC, CBD, or THCV.

[0039] Figure 3 shows the chemical reaction of decarboxylation of THCA into THC upon heating.

[0040] Figure 4 shows a table listing the major cannabinoids from Cannabis. [0041] Figure 5 shows Principal components of thylakoids: PSII (photosystem II) and its antenna pigments, Cyt. bb/f (cytochrome complex b /f). PSI (photosystem I) and its antenna pigments, NADP reductase and the ATP synthetase complex. A. With THC and/or CB bound by hydrophobic interaction. B. THC and/or CBs incorporated (or embedded) in the thylakoid membrane. C. hydrophilic molecules incorporated in the lumen of thylakoid vesicles. [0042] Figure 6 is Flow-diagram of an embodiment of a thylakoid manufacturing process.

[0043] Figure 7 is an HPLC chromatogram showing pigment profile of a stabilized thylakoid extract.

Detailed description of particular embodiments [0044] According to a particular aspect, the present invention takes advantage of the property of thylakoids to stably incorporate molecules, particularly liposoluble molecules.

[0045] Thylakoids are made of protein complexes comprising antenna pigments, photosystems I and II, reaction centers of the photosystems I and II (PSI and PSII), cytochrome b /f complex and the ATP synthetase complex (Murphy, 1986) (Figure 5). [0046] The specific organization of these thylakoids favors optimal activity of PSI and

PSII, where photochemical reactions take place. The thylakoid membrane preparations of the present invention retain photosystems properties and form vesicles that can incorporate substances of interest.

[0047] It is another object of this invention to provide products that result from the isolation of thylakoids while in the presence of Cbs. Since pure thylakoids (without Cbs) have antioxidant and anti-inflammatory activities (Bissonnette et al. , 2004; Roberts and Moreau, 2016), the final extracts containing Cbs will also demonstrate the same properties. Cbs also have antioxidant and anti-inflammatory activities (Borges et al., 2013, Giacoppo et al., 2017). By incorporating them into the thylakoid membranes, they retain these same properties. Thylakoid-Cb extracts even provides additive or synergistic activities compared to thylakoids and Cb alone.

Thylakoid vesicles

[0048] Thylakoid membranes spontaneously form microstructures (vesicles) when chloroplasts are lysed. These microstructures originate from the spontaneous reorganization of the components of the system (i.e. , stacked membrane network, the grana, non stacked network and lamellar structures). Because these microstructures are made of the proteolipidic complexes of thylakoids (Boardman ef a/., 1964; Berthold ef a/., 1981), they retain photosystem photochemical properties including photolysis and electron transport.

[0049] The terminology "thylakoid vesicles" refers herein to thylakoid membranes having a closed inner space. In particular embodiments, this inner space can incorporate hydrophilic or lipophilic molecules (Figure 5C). The membrane itself can incorporate lipophilic molecules (Figure 5B). These vesicles have an average size varying between 0.45 pm and 1.2 pm. Typical methods of preparing thylakoids will generate thylakoids vesicles. The terminology "thylakoids" without further specification refers herein to thylakoid membranes which may be in the form of vesicles or not. It thus encompasses thylakoid vesicles.

[0050] Larger fragments of thylakoids may include for instance the complete PSI and/or the PSI, and other thylakoid components. Thylakoid functional derivatives can be in the form of vesicles. Without being so limited, "thylakoid functional derivatives" can be prepared by methods described below.

[0051] As used herein the term "heterogeneous agent" refers to a molecule that is not naturally found in thylakoid vesicles and that is desirably incorporated into thylakoid vesicles in accordance with methods of the present invention. Wthout being so limited, such molecule may be a small molecule, a peptide or a protein. The present invention encompasses the use of one or more heterogeneous agents in thylakoids and/or functional derivatives of the present invention.

[0052] As used herein the terms "stably incorporated" when used in the context of heterogeneous agents stably incorporated into thylakoids vesicles of the present invention refers to agents that stay within the thylakoid vesicles for more than 10 days when refrigerated at 4°C and at least 10 days at room temperature.

Generation of reactive oxygen species

[0053] In accordance with yet another aspect, the present invention takes advantage of the ability of thylakoids and of the thylakoid functional derivatives to generate reactive oxygen species (ROS) by activation of photosystems following light absorption. Although thylakoids and their functional derivatives can be used alone, they may also advantageously incorporate other biologically active agents, such as for example, cannabinoids.

[0054] The terminology "thylakoid active/functional derivative" in its plural or singular form refers herein to thylakoid fragments that can generate reactive oxygen species through various mechanisms including charge separation (e.g., the reaction center of the P680 of PSII separates the electron donor and electron acceptor) but do not contain all components of a thylakoid per se. Minimally, these fragments contain the reaction center of the PSI or of the PSII which can induce charge separation contrarily to isolated chlorophyll perse. The reaction centers of PSI and PSII are the major generation site of ROS in thylakoids. In PSI, the primary reduced ROS is superoxide anion (Or), and its disproportionation produces H2O2 and O2. In PSII, oxygen of the ground triplet state (³Q2) is excited to singlet state (¹Q2) via charge recombination of the light-induced charge pair of the reaction center.

[0055] Thylakoids and their functional derivatives can be subjected to different treatments (e.g., treatment with urea and/or NaCI) that promote the generation ROS by affecting either the acceptor side or the donor side of the PSII (or of the PSI after activation of its ROS production with methyl viologen for example). As used herein thylakoids or their functional derivatives so treated are also encompassed within the meaning of thylakoid functional derivatives. Stabilized thylakoids as defined herein are also encompassed by the name functional derivatives. [0056] Larger fragments of thylakoids may include for instance the complete PSI and/or the PSI, and other thylakoid components. Thylakoid functional derivatives can be in the form of vesicles. Without being so limited, "thylakoid functional derivatives" can be prepared by methods described below.

[0057] Any known methods of preparing thylakoids can be used to prepare the thylakoids or functional derivatives of the present invention. In addition to the method used in Examples below, thylakoids lyophilisation can also be used (ZimmermamT et ai, 1996) . Advantageously, thylakoids used for the present invention can be isolated from any natural source of thylakoids. Hence, for instance any dark green leaf that is desirably not too fibrous or waxy can be used as a thylakoid source for the present invention. Furthermore, thylakoids of the present invention and their functional derivatives do not need preconditioning of the plant prior to extraction. For instance, it is contrary to purposes of the present invention to promote the production of antioxidants (e.g., carotenoids) in plants prior to extraction of thylakoids. On the contrary, for their application in PDT, thylakoids must generate as much ROS as possible.

[0058] Thylakoid membranes spontaneously form microstructures (vesicles) when chloroplasts are lysed. These microstructures originate from the spontaneous reorganization of the components of the system (i.e. , stacked membrane network, the grana, non stacked network and lamellar structures). Because these microstructures retain photosystem photochemical properties including photolysis and electron transport, they are likely made of the proteolipidic complexes of thylakoids (Boardman et ai, 1964; Berthold et ai, 1981).

Combination of thylakoids with liposoluble compounds

[0059] As used herein the term“heterogeneous compound” refers to a molecule that is not naturally found in thylakoid vesicles and that is desirably incorporated in thylakoids in accordance with methods of the present invention. Without being so limited, such molecule may be a of liposoluble compound. The present invention encompasses the use of one or more heterogeneous compound in thylakoids and/or functional derivatives of the present invention.

[0060] Without wishing to be bound by theory, Applicant envisages that CBs, which are lipophilic (hydrophobic) molecules tend to bind with, or be incorporated in, thylakoid membranes. When Cbs are solubilized in micelles, such as in phospholipid micelles, the micelles and thylakoid membranes, when mixed together and upon proper incubation time and agitation, incorporate with each other, thus forming Cb-containing thylakoid membranes. Of course, the person skilled in the art understands that the concentration of Cb embedded in the thylakoids (Figure 5B) can be adjusted to obtain proper ratios of Cb:thylakoids and take advantage of the biological activities of both components. W02004/046717 details the protocols for determining the threshold concentration of embedded molecules not to exceed in order to avoid negative effects on thylakoids’ electron transport activity (see Table 1).

Table 1. Threshold concentration of embedded lipid molecules before inhibiting thylakoid activity

Embedded molecule Concentration (pg/L)¹

Oleic acid 2,5

Linoleic acid 2,5

Aromatic Hydrocarbons 10

1 Detected by fluorescence inhibition of photosynthetic electron transport according to the protocol described in

W02004/046717. Threshold concentration obtained by incubating thylakoid membranes with lipid molecules for 30 min.

[0061] Alternatively, a Cb extract is solubilized in organic solvent and, upon mixing and dilution in thylakoid suspension, binds thereto by way of hydrophobic interactions (Figure 5A).

Homogenously-sourced compound

[0062] Advantageously, it is also possible to extract thylakoids from Cb-producing plants, wherein the inherent Cbs present in the photosynthetic plant parts (for example in leaves during their optimal growth period (Pacifico et al. 2007) are inherently incorporated in the resulting thylakoids, thus providing a stabilized thylakoid extract incorporating Cbs.

[0063] Applicant herein posits that liposoluble molecules such as, for example, THC and/or Cbs and other plant-extracted liposoluble molecules are incorporated in the thylakoid membranes during their extraction/isolation process, thus forming an advantageous system for the incorporation, stabilisation and carrier system of liposoluble molecules. [0064] One advantage of this aspect is provided by the simultaneous extraction of Cb and thylakoids at the same time and from the same plant parts.

[0065] A further advantage resides in the extraction and isolation of Cb in parallel from separate plant parts of the same plant, and then combination of both extracts to form a Cb- containing thylakoid formulation.

[0066] For instance, thylakoid membranes incorporate cannabinoids that, in turn, take advantage of these membranes to act as delivery systems for these plant-derived molecules (i.e. Cb from Cannabis or hemp).

[0067] Such molecules, when possessing therapeutic activity, benefits from the natural antioxidant function of the thylakoids to enhance own therapeutic activity (i.e. synergy or potentiation).

[0068] For instance, when using cannabis plants (with or without THC), the flowers are separated from the leaves and isolation of the Cbs is carried out separately from the flowers/buds whereas thylakoids are extracted from the leaves. Particularly, the trichomes present on the surface of the leaves are separated from the leaves by means of freezing and then combined with the flower/bud trichomes such that the thylakoid extraction from the leaves ONLY recovers Cbs that are present in the cannabis leaves.

Heteroqenouslv-sourced compound

[0069] Alternatively, the liposoluble compound provides from a source extraneous from the photosynthetic organism and is added to the thylakoid membranes during their extraction process.

[0070] For example, the Cbs (acid or decarboxylated forms) are obtained in an extracted / purified form as a solution or as a powder from Cbs-producing plants. The Cbs are then added to the thylakoid solution during the extraction process and combine with, or bind to, or incorporate in the membranes by hydrophobic interactions. The PSI/PSI I functional thylakoid solution is then dried and stabilized as described herein or in WO 2003/004042 and/or WO 2017/100915.

[0071] Alternatively, the Cbs are first solubilized in organic solvent before being added to the thylakoid suspension. Particularly, the Cbs are solubilized in ethanol or methanol, most particularly from 10% to 100% aqueous solvent. Particularly, the solubilized Cbs are then diluted in the thylakoid suspension at a ratio of from about 1 : 10 to about 1 : 100, thus yielding a thylakoid suspension with about 1 to 10% ethanol, thus preserving the integrity of the thylakoid PSI/PSII systems and their photosynthetic activity.

[0072] Cbs can also be solubilized in phospholipid micelles prior to being combined with thylakoids (Yang et al. 1992; Bruggemann et al. 1983). Without wishing to be bound by theory, it is posited by Applicant that phospholipids and thylakoid membranes eventually combine to at least some extent after some time and, optionally agitation, thus yielding CBs incorporated into thylakoids in a predictable ratio, depending on the initial concentrations of both lipid suspensions.

Delivery system

[0073] In accordance with an aspect of the present invention, thylakoids and their functional derivatives are used as delivery system when they incorporate lipophilic or liposoluble agents, such as therapeutic agents, for example, Cbs.

[0074] In a specific embodiment, vesicles of the present invention incorporate molecules of about 100 to 400 Da, and more specifically of about 100 to 300 Da. Vesicles of thylakoids and of their functional derivatives are advantageously of low preparation costs, are stable at least 7 to 10 days at 4 degrees and could generate fewer side effects such as allergies due to their natural origin.

[0075] Without being so limited, therapeutic agents that could advantageously be delivered by vesicles of thylakoids or of their functional derivatives include cannabinoids, such as for example:

• THC (Tetrahydrocannabinol);

• CBD (Cannabidiol);

• CBN (Cannabinol);

• CBG (Cannabigerol);

• CBC (Cannabichromene);

• CBL (Cannabicyclol);

• CBV (Cannabivarin);

• THCV (Tetrahydrocannabivarin);

• CBV (Cannabidivarin); • CBCV (Cannabichromevarin);

• CBGV (Cannabigerovarin);

• CBGM (Cannabigerol Monomethyl Ether);

• CBE (Cannabielsoin); and

• CBT (Cannabicitran).

[0076] Cannabis does not directly make the most famous cannabinoids associated with the plant, THC and Cb. Instead, it synthesizes several cannabinoid acids. These cannabinoid acids must be“activated” (decarboxylated), usually by heat, to yield the compounds that most consumers are after THC or Cb.

[0077] These classical cannabinoids are derived from their respective 2-carboxylic acids which are also known to possess some therapeutic activity:

• THCA (Tetrahydrocannabinolic acid);

• CBDA (Cannabidiolic acid);

• CBNA (Cannabinolic acid);

• CBGA (Cannabigerolic acid);

• CBCA (Cannabichromenic acid);

• CBI_A (Cannabicyclolic acid);

• CBVA (Cannabivarinic acid);

• THCVA (Tetrahydrocannabivarinic acid);

• CBVA (Cannabidivarinic acid);

• CBCVA (Cannabichrome varinic acid);

• CBGVA (Cannabigerovarinic acid);

• CBGMA (Cannabigerol acid Monomethyl ether);

• CBEA (Cannabielsoic acid); and

• CBTA (Cannabicitranic acid).

[0078] When cannabinoid acids are exposed to heat energy, they lose the“A” part and turn into neutral, rather than acidic, plant cannabinoids (Figure 2). Following decarboxylation (catalyzed by heat, light, or alkaline conditions), each of the cannabinoid acids yields a corresponding cannabinoid compound.

[0079] Cannabinoid acids are not known to produce intoxicating effects like THC. But they do have a variety of interesting properties. For example, many cannabinoid acids have antibiotic or insecticidal properties. This is likely related to the reason Cannabis produces these compounds in the first place: to defend itself.

[0080] But in addition to THCA and CBDA, there are number of related cannabinoid acids that can be produced by Cannabis. Other cannabinoids present in cannabis are also known the have functional health benefits such as, for example:

Table 2. Other classes of natural cannabinoids

Methods of preparinq/extractinq thylakoids

[0081] Any known methods of preparing thylakoids can be used to prepare the thylakoids or functional derivatives of the present invention. In addition to the method used in Examples below, thylakoids lyophilisation can also be used (Zimmerman et al. , 1996). [0082] Advantageously, thylakoids used for the present invention can be isolated from any natural source of thylakoids. Hence, for instance any dark green leaf that is desirably not too fibrous or waxy can be used as a thylakoid source for the present invention.

[0083] Furthermore, thylakoids of the present invention and their functional derivatives do not need preconditioning of the plant prior to extraction. For instance, it is not necessary for purposes of the present invention to promote the production of antioxidants (e.g., carotenoids) in plants prior to extraction of thylakoids.

[0084] Alternatively, when it is desired to use cannabis leaves to extract thylakoids, it is possible to condition the leaves to remove the trichomes with dry ice (Yerger et al. 1992), prior to extraction. Acid vs decarboxylated forms of Cbs

[0085] Cannabinoids are known to be in their acid form when extracted under room temperature conditions. It is commonly accepted that Cbs possess the health virtues when present in their decarboxylated state (Wang et al. 2016). However, it is also known that Cbs can have some beneficial health effects even in their acid form (Nadal et ai, 2017). [0086] It is therefore an object of the invention to provide a functional thylakoid extract that comprises Cbs in their acid form.

[0087] It is also an object of the invention to provide a functional thylakoid extract that comprises Cbs in their decarboxylated form.

[0088] In order to achieve a combination of thylakoids with decarboxylated Cbs, decarboxylation must take place before contacting with thylakoids to avoid disrupting the PSI/PSI I system that provides functionality of the thylakoid membranes. Such decarboxylation thus may take place during Cb extraction by heating at a temperature allowing decarboxylation as well known in the art Wang et al. 2016). On the other hand, decarboxylation may take place immediately before adding the Cbs to the thylakoid solution (Wang et al., 2016).

Stabilized extract

[0089] Particularly, the thylakoid extract is stabilized in its fundamental state (i.e. stable) by being devoid of any electron donor (such as water). More particularly, the extract is stabilized by containing no more than 10% of water (or other electron donor), particularly less than 10% water.

[0090] Most particularly, the composition is defined as a raw organic plant, active thylakoid extract, wherein the ratio chlorophyll a to total pigment is at least 0.4, particularly at least 0.5, more particularly at least 0.6. In particular, the pigment comprised in the thylakoid extract is selected from the group consisting of chlorophyll a, chlorophyll b, and carotenoids. More particularly, the pigment comprised in the thylakoid extract is selected from the group consisting of chlorophyll a, chlorophyll b, lutein, and optionally, b-carotene and/or pheophytin. Still, most particularly, the pigment comprised in the thylakoid extract consists essentially of chlorophyll a (more than 40%), followed by chlorophyll b (about 10 -15%), lutein (about 10% or less), b-carotene (about 3%) and pheophytin (less than 1 %).

[0091] The extract may be in powder form, such as a lyophilized dried composition, or may be admixed with physiologically-acceptable solid or liquid excipients, such as: PEG or PVP DMSO to form a stabilized solution of suspension, inasmuch as the composition remains free, or substantially free, of electron donors (such as water), to maintain activity of the functional pigments.

[0092] Particularly, the stabilized extract is in solid form, more as particularly, as a powder. Still, more particularly, the extract is in powder form with at least 25 mg pigments per gram of powder.

[0093] Particularly, the thylakoid extract is a Cannabis or Hemp thylakoid extract, and more particularly extracted from Cannabis or Hemp leaves, flowers, and other plant parts that contain chloroplasts and/or Cbs.

[0094] More particularly, the whole plant or the whole leaves are harvested for extraction. Most particularly, the plant parts can be chosen from: lead shoots, stem from vegetative plant, de-leafed flowers, full-dressed flowers, full plant from flowering plant; leaves from flowering plant; root from flowering plant; and/or stem from flowering plant. Particularly, for producing a extract that is devoid or low in THCA, extraction is carried out with cannabis leaves, more particularly“fan” leaves.

Cannabis varieties

[0095] According to a particular aspect, the method as defined herein is carried out with cannabis selected from: Cannabis Sativa, Cannabis Indica or Hemp.

[0096] Depending on the pharmacological profile desired for the juice, several cannabis varieties can be chosen: For example, Hemp, Cannabis Indica, Cannabis Sativa and/or Hybrids can be used (see Table 3 for specific strains). Table 3. Approximate THC and CB levels of some premium cannabis strains.

[0097] According to a particular aspect, the THC and/or Cb and/or thylakoids of the present invention may be isolated from photosynthetic organisms that naturally produce THC, CB as desired.

[0098] Alternatively, the thylakoids of the present invention may be isolated from any photosynthetic organism, whether or not it naturally produces THC and/or Cb.

[0099] In a further aspect, the THC and/or Cb and/or thylakoids of the present invention may be isolated from photosynthetic organisms that are genetically modified (GMO) to produce (or to produce more, or to produce less) THC and/or Cb, compared to its natural (i.e. non-GMO) counterpart.

Composition

[00100] The invention also provides pharmaceutical compositions which comprise THC and/or CB incorporated into thylakoid membranes or vesicles. Particularly, the thylakoid vesicles are active/functional photosynthetically and can act as antioxidants.

[00101] The invention also provides a pharmaceutical composition (medicament) comprising at least the thylakoid vesicles of the invention {e.g., incorporating or not at least one heterogeneous compound), and a pharmaceutically acceptable diluent, carrier, salt or adjuvant. Such carriers include, for example, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof. The pharmaceutical composition may be adapted for the desired route of administration (e.g., oral, parental, intravenous, intramuscular, intraperitoneal, aerosol).

[00102] More particularly, the raw powder may be compressed in tablets, encapsulated, or packaged into aliquot packets or pouches. Alternatively, the powder may be mixed with suppository excipients and molded to form suppositories.

[00103] The invention also provides pharmaceutical compositions which comprise one or more cannabinoid(s). In accordance with a particular aspect, the invention describes a composition comprising an effective amount of an active thylakoid extract incorporating an active amount of a liposoluble compound. [00104] Particularly the composition comprises purified functional photosynthetic pigments, in addition to a cannabinoid, incorporated in a functional thylakoid membrane environment.

[00105] Still particularly, the functional photosynthetic pigments are quiescent and can be activated photosynthetically. More particularly, the functional photosynthetic pigments are stabilized in their fundamental state (i.e. stable) by being devoid of any electron donor (such as water).

Therapeutic indications

[00106] According to a particular embodiment, there is provided the use of the composition as defined herein, for the maintenance of health. Particularly, according to a particular embodiment, there is provided a use of the composition as defined herein for pharmaceutical, cosmetical and/or cosmeceutical uses.

[00107] It is well known that cannabinoids in general possess anti-inflammatory, antioxidant and analgesic activities. As well, purified thylakoids possess have antioxidant and anti-inflammatory activities, such as for example, for fighting against inflammatory bowel diseases such as ulcerative colitis and Crohn’s disease, and cardiovascular diseases; or for helping or prevent skin aging, wrinkles, skin hydration and/or skin UV protection.

[00108] It is therefore herein provided a use of the present composition comprising CB in thylakoid vesicles for the prevention and/or treatment defined herein or other conditions known to be addressed by one or the other of these components.

[00109] According to a further embodiment, the present invention provides the use of the composition as defined herein, as an antioxidant, appetite stimulant and/or anti-inflammatory.

[00110] More particularly, the composition may be used for relaxation, anxiolytic, anti depressor and/or pain relief purposes.

[00111] More particularly, the composition may be used for neuroprotection.

[00112] More particularly, the composition may be used for the prevention or treatment of Alzheimer’s disease, multiple sclerosis, Parkinson’s disease, PTSD and Crohn’s disease. More particularly, the composition may be used for its antispasmodic effects to minimize spasms or convulsions, such as those in epilepsy. [00113] More particularly, the composition may be used for mitigating the side-effects of chemotherapy since it helps minimize vomiting and nausea, making it useful for cancer patients undergoing chemotherapy.

[00114] Alternatively, the composition may be used in the treatment of prevention of cancer.

Table 4. Conditions potentially responding to cannabinoid therapies

[00115] Although the present invention has been described hereinabove by way of specific embodiments thereof, it can be modified, without departing from the spirit and nature of the subject invention as defined in the appended claims. [00116] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

EXAMPLES

Example 1. Preparation of thylakoids (Figure 6)

[00117] I: Homogenisation: All steps were conducted in the dark or under green light and at cold temperature (samples on ice or procedure in cold room). A hundred grams of deveined spinach leaves were crushed in a mixer with 350 mL of homogenizing buffer comprising TES- NaOH 20mM pH 7.5, sorbitol 330mM and MgC 5mM. Other conventional buffers could be used including TES, Hepes, Tris, phosphate, tricine and MOPs. The homogenate was filtered on a cheese cloth and the filtrate was centrifuged 2 min at 2500X g at 4°C on a Eppendorf™5810-R, rotor # A-4-44.

[00118] II: Chloroplasts lysis: The pellet was then resuspended in 200 mL of hypotonic solution consisting of the homogenizing buffer diluted in water 1 : 20 v/v (i.e. TES-NaOH 20mM pH 7.5, sorbitol 330mM and MgCL2 5mM diluted 1 :20 v/v with water). This step was used to lyse the chloroplast membranes. The resulting solution was then crushed in a Wheaton™ mixer and centrifuged 3 min. at 3500Xg at 4°C on a Eppendorf™5810-R, rotor* A-4-44.

[00119] III: Resuspension: The resulting pellet contained the purified thylakoids. They were then resuspended in a buffer consisting of TES-NaOH 20 mM pH 7.5, sorbitol 330 mM, MgCI_₂ 5 mM and NH4CI 1 mM to obtain a final concentration of chlorophyll between 2 and 3 mg/ml_. The chlorophyll concentration was assessed according to the method taught in Porra (1989) described herein. The solution was then either stored at -80°C for later use or was stabilized as described in Example 3 below for storing at room temperature. The thylakoids obtained at the end of this process were in the form of vesicles. Example 2. Combination of Cb with thylakoids

[00120] A specific example of the above-mentioned separating step comprises centrifuging the first extract for 10 minutes at 10 000 g in a tube equipped with a filter in a superior portion of the tube, the filter having a suitable porosity onto which cell debris and membranes deposit while the thylakoids and the liquid phase pass through the filter, the thylakoids forming a pellet in an inferior portion of the tube. Alternatively, gross purification is achieved by recovering cell debris and membranes first by pressing and/or filtering, for example, followed by a finer purification, e.g. separating thylakoids from the liquid phase.

[00121] Right before or after separation of plant debris, cannabinoids are added to the solution containing the thylakoid membranes. To do this, a solution of Cbs (maximal concentration: 1000 mg / ml_) is prepared by dissolving the Cbs in 100% methanol or ethanol. This latter solution is then diluted in the same buffer as before (final concentration of ethanol or methanol 10% v/v). This diluted solution of Cb is added to the thylakoid buffered solution by gentle stirring for 30 minutes.

[00122] The solution is then dried in the usual manner to stabilize the Cb- containing thylakoid extract. It is important

Example 3. Stabilization of thylakoid extract

[00123] Water is eliminated under vacuum freeze drying or by exchanging it against a non-denaturing amphoteric solvent or surfactant after step c), non-denaturing meaning not capable of dissociating or of damaging the thylakoid structural components.

[00124] After water elimination, thylakoid extracts containing Cbs are stabilized by adding the following step d): eliminating any electron donor from said extracts so as to inactivate and stabilize the photosynthetic pigments preferably in the presence of sugars (which may protect components against cold).

[00125] The first contemplated electron donor is water, so the extracts are processed to be water-free. Amphoteric solvents which have been tried with success are propylene glycol (PEG) and polyvinylpyrrolidone (PVP) (WO 2001/049305; WO 2017/100915).

[00126] Thylakoids of the present invention may be stabilized using the method described in WO 2001/049305 or WO 2017/100915, as described below or by other stabilization methods (e.g. lyophilisation). A preparation of 2 to 3 mg/ml_ of thylakoids were mixed with a pH 6.5 phosphate buffer containing: 0.02 M phosphate, sucrose 300 mM, NH₄CI 10 mM, MgCI_₂ 20 mM, EDTA 10 mM, polyvinylpyrrolidine (PVP) (125 pl_ of solution 20% for 1 ml_) and liposomes made of phosphatidylcholine and phosphatidylglycerol in a ratio of 10 mg/ml_: 1 mg mU to obtain a solution of 0.0125 mg of liposome/mL of chlorophyll/thylakoid membranes buffered solution. In this stabilized thylakoid membrane formulation, the final concentrations of PVP is of 2% v/v, the final concentration of liposomes is of 0.0125 mg/ml_ and that of chlorophyll is of 0.125mg/ml_

[00127] The PVP increases the stability of the thylakoid preparations as compared to the same preparation without PVP but a useful stability was nevertheless obtained even without PVP. The amount of PVP may be varied without affecting the stability of the thylakoid membranes between about 0 and about 4% PVP v/v (data not shown). Other conventional buffers at pH ranging between about 6.2 and about 7.8 -a variation of about 10% is generated within this pH range (data not shown) - can also be used without affecting the usefulness of the thylakoid preparation of the present invention.

[00128] The quantity of thylakoid liposomes used was determined via the ratio chlorophyll/liposome. This ratio may vary and is preferably at least equal to 10 mg/ml_ : 1 mg/ml_ and better results with regards to stability and sensitivity were obtained with a ratio 100 mg/ml: 1 mg/ml_. Any ratio higher than 100:1 is expected to work. The liposomes are believed to help to the formation of thylakoid membranes vesicles thereby increasing their stability and increasing the consistency of the readings. The quantity used can be varied as long as the final ratio of chlorophyll/liposomes is at least about 10 mg/ml_: 1 mg/ml_.

[00129] One hundred (100) mI_ of this solution was poured into a 5 ml_ or 7 ml_ tube previously refrigerated. The tubes were then frozen and then lyophilised with a speed-vac™ from Savant (#SS-22). Lyophilisation was conducted 4 hours in order to remove water completely from the 100 pL amount. The tubes were then closed and kept at the desired temperature.

Example 4. Preparation of CB-containing thylakoids from Cannabis

[00130] Step I of the preparation of thylakoids was as described in Example 1 , except that cannabis fan leaves are used instead of spinach. [00131] Cannabinoids are found in lipophilic glandular structures called trichomes (Kim and Mahberg, 1997). By their basic lipophilicity, these structures are extracted in a manner similar as the thylakoid membranes when extracted in aqueous buffer solution under controlled conditions.

[00132] The method may further comprise the following step c): separating thylakoids, cell debris/membranes and liquid phase from each another, to form a second, third and fourth extracts essentially constituted by 2) isolated thylakoids and Cb, 3) cell debris and membranes, and 4) liquid phase, respectively. This step of separating may be performed by sedimentation coefficient on centrifugation on sucrose phase.

[00133] Alternatively, a specific example of such separating step comprises centrifuging the first extract for 10 minutes at 10 000 g in a tube equipped with a filter in a superior portion of the tube, the filter having a suitable porosity onto which cell debris and membranes deposit while the thylakoids and the liquid phase pass through the filter, the thylakoids forming a pellet in an inferior portion of the tube. Alternatively, gross purification may be achieved by recovering cell debris and membranes first by pressing and/or filtering, for example, followed by a finer purification, e.g. separating thylakoids from the liquid phase.

[00134] The final extract is then comprised of a mixture of thylakoids and cannabinoids in a ratio that optimally retains the antioxidant and anti-inflammatory properties that thylakoids and cannabinoids separately possess.

Example 5. Separate extractions of CBs and thylakoids from the same cannabis plant Extraction of cannabinoids

[00135] The flowers/buds are separated manually from the leaves. Optionally, the trichomes may be removed from the leaves and combined with the flowers/buds for Cbs extraction. Flowers/buds (with or without leave’s trichomes) are ground and homogenized and the Cbs are extracted as well known in the art (Sexton et al., 2017). Optionally, the Cbs are decarboxylated under heat according to protocols well known in the art (Wang et al., 2016).

[00136] The CBs are then solubilized in 100% ethanol or methanol for further incorporation with thylakoids. Extraction of thylakoids

[00137] The remaining leaves (with or without the trichomes) are crushed and homogenized and the thylakoids are extracted as described above.

Example 6. Solubilization of CB in phospholipids prior to adding to thylakoids

[00138] It has been established that phospholipids, when incorporated in thylakoids, increase their stability (WO 2004/046717). It is thus posited that, Cbs are incorporated in thylakoids by way of phospholipid micelles and act in a two-fold manner, by stabilizing the thylakoids, and by incorporating biologically active Cb that may potentiate the biological activity of thylakoids.

Preparation of liposomes containing cannabinoids

[00139] Alternatively, the Cbs are incorporated in phospholipids membranes or micelles (according to WO 2004/046717) prior to mixing with the thylakoids. Liposomes or micelles consist of a lipid mixture of phosphatidylcholine and phosphatidylglycerol in a ratio of 10: 1.

[00140] Phosphatidylcholine and phosphatidylglycerol in a ratio of 10: 1 in powder form are dissolved in ethanol in the presence of powdered cannabinoids (up to 1000 mg / ml) or mixed with Cb previously dissolved in ethanol. For forming the micelles, the ethanol is then evaporated using a roto-evaporator.

[00141] After ethanol removal, the thylakoid homogenization or resuspension buffer is then added to the remaining lipid deposit, thus forming a suspension. Liposomes are then formed by subjecting the suspension to 3 cycles of freeze-thaw (liquid nitrogen 5 min) / thawing (50° C, 15 min) or rotating in a roto-evaporator for 24 hours at room temperature.

Incorporation of cannabinoids in thylakoids

[00142] The two suspensions (Cb micelles and thylakoids) are then combined in a predetermined ratio to achieve a phospholipid concentration that does not affect/disrupt the PS activity of the thylakoid membranes. They are then mixed by gentle agitation. Once again, the procedure is carried out under controlled temperature conditions at 4C and green or dark light.

[00143] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features herein before set forth, and as follows in the scope of the appended claims.

[00144] All patents, patent applications and publications mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent, patent application or publication was specifically and individually indicated to be incorporated herein by reference.

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Claims

1. A composition comprising one or more cannabinoids (Cb) stably incorporated in a thylakoid extract.

2. A composition comprising a thylakoid membrane incorporating a lipophilic compound.

3. The composition of claim 1 or 2, wherein the lipophilic compound is stably embedded in the thylakoid membrane.

4. The composition of claim 1or 2, wherein the lipophilic compound is bound to the thylakoid membrane.

5. The composition of claim 1 or 2, wherein the lipophilic compound is incorporated in the thylakoid vesicles.

6. The composition of claim 1 to 5, wherein the thylakoid extract, membrane or vesicles have the capacity to be activatable in presence of electron donors.

7. The composition of claim 1 to 6, wherein the lipophilic molecule is obtained from a cannabinoid (Cb)-producing plant, such as cannabis sativa, cannabis indica or hemp.

8. The composition of claim 1 to 7, wherein the thylakoid extract, membrane or vesicles are obtained from a cannabinoids (Cb)-producing plant, such as cannabis sativa, cannabis indica or hemp.

9. The composition of claim 1 or 8, wherein the Cb is selected from the group consisting of: THC (Tetrahydrocannabinol); CB (Cannabidiol); CBN (Cannabinol); CBG (Cannabigerol); CBC (Cannabichromene); CBL (Cannabicyclol); CBV (Cannabivarin); THCV (Tetrahydrocannabivarin); CBV (Cannabidivarin); CBCV (Cannabichromevarin); CBGV (Cannabigerovarin); CBGM (Cannabigerol Monomethyl Ether); CBE (Cannabielsoin); CBT (Cannabicitran); THCA (Tetrahydrocannabinolic acid); CBA (Cannabidiolic acid); CBNA (Cannabinolic acid); CBGA (Cannabigerolic acid); CBCA (Cannabichromenic acid); CBLA (Cannabicyclolic acid); CBVA (Cannabivarinic acid); THCVA (Tetrahydrocannabivarinic acid); CBVA (Cannabidivarinic acid); CBCVA (Cannabichrome varinic acid); CBGVA (Cannabigerovarinic acid); CBGMA (Cannabigerol acid Monomethyl ether); CBEA (Cannabielsoic acid); and CBTA (Cannabicitranic acid).

10. The composition according to any one of claims 1 or 9, in dry form or in a medium composed of an amphoteric solvent such as propylene glycol or PVP.

11. A pharmaceutical composition comprising the Cb-containing thylakoid extract of any one of claims 1 to 9, in admixture with a pharmacologically acceptable carrier.

12. Thylakoids or thylakoid functional derivatives for use as carriers for lipophilic compounds.

13. The thylakoids or thylakoid functional derivatives for use according to claim 12, wherein said lipophilic compounds are cannabinoids (Cb).

14. The thylakoids or thylakoid functional derivatives for use according to claim 13, wherein the Cb is selected from the group consisting of: THC (Tetrahydrocannabinol); CB (Cannabidiol); CBN (Cannabinol); CBG (Cannabigerol); CBC (Cannabichromene); CBL (Cannabicyclol); CBV (Cannabivarin); THCV (Tetrahydrocannabivarin); CBV (Cannabidivarin); CBCV (Cannabichromevarin); CBGV (Cannabigerovarin); CBGM (Cannabigerol Monomethyl Ether); CBE (Cannabielsoin); CBT (Cannabicitran); THCA (Tetrahydrocannabinolic acid); CBA (Cannabidiolic acid); CBNA (Cannabinolic acid); CBGA (Cannabigerolic acid); CBCA (Cannabichromenic acid); CBI_A (Cannabicyclolic acid); CBVA (Cannabivarinic acid); THCVA (Tetrahydrocannabivarinic acid); CBVA (Cannabidivarinic acid); CBCVA (Cannabichrome varinic acid); CBGVA (Cannabigerovarinic acid); CBGMA (Cannabigerol acid Monomethyl ether); CBEA (Cannabielsoic acid); and CBTA (Cannabicitranic acid).

15. Use of thylakoids or thylakoid functional derivatives as carriers for lipophilic compounds.

16. The use according to claim 15, wherein said lipophilic compounds are cannabinoids (Cb).

17. The use of claim 16, wherein the Cb is selected from the group consisting of: THC (Tetrahydrocannabinol); CB (Cannabidiol); CBN (Cannabinol); CBG (Cannabigerol); CBC (Cannabichromene); CBL (Cannabicyclol); CBV (Cannabivarin); THCV (Tetrahydrocannabivarin); CBV (Cannabidivarin); CBCV (Cannabichromevarin); CBGV (Cannabigerovarin); CBGM (Cannabigerol Monomethyl Ether); CBE (Cannabielsoin); CBT (Cannabicitran); THCA (Tetrahydrocannabinolic acid); CBA (Cannabidiolic acid); CBNA (Cannabinolic acid); CBGA (Cannabigerolic acid); CBCA (Cannabichromenic acid); CBLA (Cannabicyclolic acid); CBVA (Cannabivarinic acid); THCVA (Tetrahydrocannabivarinic acid); CBVA (Cannabidivarinic acid); CBCVA (Cannabichrome varinic acid); CBGVA (Cannabigerovarinic acid); CBGMA (Cannabigerol acid Monomethyl ether); CBEA (Cannabielsoic acid); and CBTA (Cannabicitranic acid).

18. A method for producing a Cb-comprising thylakoid extract comprising the steps of combining a Cb extract with a thylakoid suspension.

19. A method for producing a Cb-comprising thylakoid extract comprising the steps of extracting thylakoids from a Cb-producing plant part having photosynthetic activity, under conditions adapted to recover Cbs in said thylakoid extract.

20. The method of claim 18 or 19, wherein the Cb and/or thylakoids are obtained from a natural photosynthetic organism.

21. The method of claim 18 or 19, wherein the Cb and/or thylakoids are obtained from a genetically modified photosynthetic organism.

22. The method according to any one of claims 18 to 21 , wherein the Cb is selected from the group consisting of: THC (Tetrahydrocannabinol); CB (Cannabidiol); CBN (Cannabinol); CBG (Cannabigerol); CBC (Cannabichromene); CBL (Cannabicyclol); CBV (Cannabivarin); THCV (Tetrahydrocannabivarin); CBV (Cannabidivarin); CBCV (Cannabichromevarin); CBGV (Cannabigerovarin); CBGM (Cannabigerol Monomethyl Ether); CBE (Cannabielsoin); CBT (Cannabicitran); THCA (Tetrahydrocannabinolic acid); CBA (Cannabidiolic acid); CBNA (Cannabinolic acid); CBGA (Cannabigerolic acid); CBCA (Cannabichromenic acid); CBLA (Cannabicyclolic acid); CBVA (Cannabivarinic acid); THCVA (Tetrahydrocannabivarinic acid); CBVA (Cannabidivarinic acid); CBCVA (Cannabichrome varinic acid); CBGVA (Cannabigerovarinic acid); CBGMA (Cannabigerol acid Monomethyl ether); CBEA (Cannabielsoic acid); and CBTA (Cannabicitranic acid).

23. Use of the composition as defined in any one of claims 1 to 14, for the maintenance or prevention of conditions requiring anti-inflammatory, antioxidant and analgesic activities.

24. Use of the composition as defined in any one of claims 1 to 14, for treating or preventing skin aging, wrinkles, skin hydration and/or skin UV damage.

25. The use of claim 23, wherein said condition is selected from: inflammatory bowel diseases, such as ulcerative colitis and Crohn’s disease, and cardiovascular diseases.

26. Use of the composition as defined in any one of claims 1 to 14, as an anxiolytic, anti-depressor and/or for pain relief purposes.

27. Use of the composition as defined in any one of claims 1 to 14, for neuroprotection or as antispasmodic effects to minimize spasms or convulsions, such as those in epilepsy.

28. Use of the composition as defined in any one of claims 1 to 14, for the prevention or treatment of a disease selected from the group consisting of: Alzheimer’s disease, multiple sclerosis, Parkinson’s disease, post-traumatic stress syndrome (PTSD) and Crohn’s disease.

29. Use of the composition as defined in any one of claims 1 to 14, for mitigating the side-effects of chemotherapy such as vomiting and nausea.

30. Use of the composition as defined in any one of claims 1 to 14, for the treatment or prevention of cancer.

31. A method for the treatment or prevention of a condition caused by inflammation, oxidative stress, comprising administering, to a subject suffering therefrom, an effective amount of a composition as defined in any one of claims 1 to 14.

32. A method for the treatment or prevention of skin aging, wrinkles, skin dehydration and/or skin UV damage, comprising administering to a subject suffering therefrom, an effective amount of the composition as defined in any one of claims 1 to 14.

33. The method of claim 31 , wherein said condition is selected from: inflammatory bowel diseases, such as ulcerative colitis and Crohn’s disease, and cardiovascular diseases.

34. A method for the treatment or prevention anxiety, depression, and/or pain comprising administering to a subject suffering therefrom, an effective amount of the composition as defined in any one of claims 1 to 14.

35. A method for neuroprotection or as antispasmodic to minimize spasms or convulsions, such as those in epilepsy anxiety, depression, and/or pain comprising administering to a subject suffering therefrom, an effective amount of the composition as defined in any one of claims 1 to 14.

36. A method for the prevention or treatment of a disease selected from the group consisting of: Alzheimer’s disease, multiple sclerosis, Parkinson’s disease, post-traumatic stress syndrome (PTSD) and Crohn’s disease, comprising administering, to patient suffering therefrom, an effective amount of the composition as defined in any one of claims 1 to 14.

37. A method for mitigating side-effects of chemotherapy such as vomiting and nausea comprising administering to a subject suffering therefrom, an effective amount of the composition as defined in any one of claims 1 to 14.

38. A method for the treatment or prevention of cancer comprising administering to a subject suffering therefrom, an effective amount of the composition as defined in any one of claims 1 to 14.