Prenylated flavone compounds

Abstract

Disclosed herein are prenylated flavone compounds having the structure of Formula (I), or a pharmaceutically acceptable salt and / or hydrate or solvate thereof. Also disclosed herein are methods for producing prenylated flavone compounds, and uses for such compounds, including in medical treatments.

Description

Prenylated flavone compounds CROSS REFERENCE

[0001] The present application derives priority from Australian provisional patent application no. 2024903376, filed 18 October 2024, the entire contents of which is incorporated herein by cross reference in its entirety. FIELD OF THE INVENTION

[0002] The invention relates to prenylated flavone compounds, methods for producing prenylated flavone compounds, and uses for such compounds. However, it will be appreciated that the invention is not limited to this particular field of use. BACKGROUND

[0003] The following discussion of the prior art is provided to place the invention in an appropriate technical context and enable the advantages of it to be more fully understood. It should be appreciated, however, that any discussion of the prior art throughout the specification should not be considered as an express or implied admission that such prior art is widely known or forms part of the common general knowledge in the field.

[0004] Cannflavins are a small family of prenylated chrysoeriol flavone molecules from Cannabis sativa that have attractive antioxidant and anti-inflammatory properties. Three naturally occurring cannflavins have been identified to-date (Figure 1). The most abundant of these are cannflavin A and cannflavin B, where chrysoeriol is prenylated at the 6-position with geranyl diphosphate (GPP) or dimethylallyl diphosphate (DMAPP), respectively. A minor cannflavin has also been identified in extracts of C. sativa where the 8-position of chrysoeriol is geranylated (cannflavin C). Chemical synthesis of a structural isomer of cannflavin B has also been reported where the dimethylallyl group occurs at the 8-position (isocannflavin B or caflanone), but this structure has not been identified in plant extracts to-date.

[0005] The final step in biosynthesis of cannflavin A and cannflavin B is catalysed by a prenyltransferase enzyme, CsPT3, that can ligate either geranyl diphosphate or dimethylallyl diphosphate to chrysoeriol at the 6-position to yield cannflavin A or cannflavin B. The concentration of cannflavin C in hemp biomass is less than 0.01% of the cannflavin A content. CsPT3 has also been shown to be able to ligate geranyl diphosphate or dimethylallyl diphosphate to apigenin.

[0006] In nature, the CsPT3 enzyme is localised in the chloroplast. This localisation limits the range of prenyl diphosphate co-substrates that are available to the enzyme. Two distinctmetabolic pathways for synthesis of prenyl diphosphates occur in plants: the mevalonate pathway in the cytosol, and methyl erythritol diphosphate pathway in the plastid. Both pathways synthesise the universal isoprenoid precursor molecules, DMAPP and its structural isomer, isopentenyl diphosphate (IPP). However, the cytosol and the plastid also have different sets of enzymes for downstream utilization of these 5-carbon precursor molecules, and a consequence is that GPP is only found in the chloroplast while the fifteen-carbon farnesyl diphosphate (FPP) is only found in the cytosol (Figure 2). In the cytosol, 1 molecule of DMAPP and 2 molecules of IPP are conjugated to produce FPP, which is a precursor to sterol biosynthesis. The ten carbon GPP intermediate is not released from the active site of the FPP synthase enzyme in significant quantities. In the plastid, 1 molecule of DMAPP and 1 molecule of IPP are conjugated to produced GPP, which can also be conjugated to a second GPP to produce the 20-carbon geranylgeranyl diphosphate (GGPP). FPP does not occur in the plastid, and therefore prior to the present work it was unknown whether the CsPT3 enzyme could accept FPP as a co-substrate to produce an alternative cannflavin molecule with a 15-carbon prenyl tail (referred to herein as cannflavin-15, Cfl15).

[0007] Prenylated flavones, such as cannflavins, have also been shown to be promising candidates for treating a variety of cancers, including transitional cell carcinoma, bladder, brain, breast, Kaposi sarcoma, leukemia, lung, melanoma, ovarian, pancreatic, colon, and prostate cancer.

[0008] It is an object of the present invention to overcome or ameliorate one or more the disadvantages of the prior art, or at least to provide a useful alternative. SUMMARY OF THE INVENTION

[0009] The inventors of the present application have surprisingly discovered that by using a prenyl transferase enzyme, it is possible to form novel prenylated flavone molecules, in particular having three or more prenyl units.

[0010] In a first aspect of the invention there is provided a compound having the structure of Formula (I):,or a pharmaceutically acceptable salt and / or hydrate or solvate thereof; wherein: R1is H or OCH3; R2is tri-prenyl, tetra-prenyl, penta-prenyl, hexa-prenyl, hepta-prenyl, or octa-prenyl; and R3is H.

[0011] The following options may be used in conjunction with the first aspect, either individually or in any combination.

[0012] In certain embodiments, R2is tri-prenyl, tetra-prenyl, penta-prenyl, hexa-prenyl, hepta- prenyl, or octa-prenyl. In certain embodiments, R2is tri-prenyl, tetra-prenyl, penta-prenyl, or hexa-prenyl. In certain embodiments, R2is tri-prenyl, tetra-prenyl, penta-prenyl, hexa-prenyl, hepta-prenyl, or octa-prenyl; and R1is OCH3. In certain embodiments, R2is tri-prenyl, tetra- prenyl, penta-prenyl, or hexa-prenyl; and R1is OCH3.

[0013] In certain embodiments, R2is all-trans tri-prenyl, all-trans tetra-prenyl, all-trans penta- prenyl, all-trans hexa-prenyl, all-trans hepta-prenyl, or all-trans octa-prenyl. In certain embodiments, R2is all-trans tri-prenyl, all-trans tetra-prenyl, all-trans penta-prenyl, or all-trans hexa-prenyl. In certain embodiments, R2is all-trans tri-prenyl, all-trans tetra-prenyl, all-trans penta-prenyl, all-trans hexa-prenyl, all-trans hepta-prenyl, or all-trans octa-prenyl; and R1is OCH3. In certain embodiments, R2is all-trans tri-prenyl, all-trans tetra-prenyl, all-trans penta- prenyl, or all-trans hexa-prenyl; and R1is OCH3.

[0014] In certain embodiments, R1is OCH3.

[0015] In certain embodiments, R2is tri-prenyl, tetra-prenyl, penta-prenyl, or hexa-prenyl.

[0016] In certain embodiments, R2is tri-prenyl, or tetra-prenyl.

[0017] In certain embodiments, R2is tri-prenyl, or tetra-prenyl.

[0018] In certain embodiments, R2is tri-prenyl, or tetra-prenyl; and R1is OCH3.

[0019] In certain embodiments, the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof has a structure selected from the following:, and pharmaceutically acceptable salts and / or hydrates or solvates thereof.

[0020] In certain embodiments, the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof has the following structure:acceptable salt and / or hydrate or solvate thereof.

[0021] The compounds of the invention may exist in both unsolvated and solvated forms. The term ‘solvate’ is used herein to describe a molecular complex comprising the compound of the invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules, for example, ethanol. The term ‘hydrate’ is employed when the solvent is water.

[0022] It will be appreciated by the person of skill in the art that the compounds of the first aspect will, due to the poly-prenyl group, present as geometric isomers, such as cis / trans and E / Z isomers. While drawn in one configuration herein for the sake of convenience, it should be appreciated that the double bonds of all compounds of the first aspect may be in the E or Z form and the double bond of every structure drawn herein is explicitly considered to be represented in both the E and Z isomeric forms. Synthesis of the compounds of the first aspect may result in substantially pure forms of E or Z isomer or a mixture of E and Z forms with respect to anydouble bond contained therein, which forms may be used in any of the methods and applications described herein in that particular form. Similarly, it will be appreciated that in any aspect of the present invention when compounds of the first aspect are provided in a composition or formulation then each compound may be present with any double bond in either substantially the E or substantially the Z isomeric form or may be present as a mixture of both. Compositions

[0023] In a second aspect of the invention there is provided a pharmaceutical composition comprising the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect and a pharmaceutically acceptable excipient.

[0024] The following options may be used in conjunction with the second aspect, either individually or in any combination.

[0025] In certain embodiments, the composition is suitable for topical administration.

[0026] The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect may be formulated as a dietary supplement or a pharmaceutical composition. Such compositions may comprise about 0.00001% to about 99% by weight of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect and any range in between. For example, typical doses may comprise from about 0.1 μg to about 100 μg of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof described herein per 300 mg dose, such as about 0.5 μg, about 1 μg, about 2 μg, about 3 μg, about 4 μg, about 5 μg, about 6 μg, about 7 μg, about 8 μg, about 9 μg, about 10 μg, about 25 μg, about 50 μg, or about 75 μg per 300 mg dose, such as from about 0.1 μg to about 10 μg, or from about 1 μg to about 5 μg, or from about 1 μg to about 2 μg per 300 mg dose (and all related increments and percentages by weight).

[0027] The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect described herein may be used in any suitable amount, such as in doses comprising from about 1 to about 10000 ng / kg, such as from about 1 to about 1000, about 1 to about 500, about 10 to about 250, or about 50 to about 100 ng / kg, such as about 1, about 10, about 25, about 50, about 75, about 100, about 150, about 200, about 250, about 300, or about 500 ng / kg. Similar amounts, higher amounts, or lower amounts may be used for administration.

[0028] Compositions herein comprise one or more compounds provided herein. The compounds are, in one embodiment, formulated into suitable pharmaceutical preparations such as solutions, suspensions, tablets, creams, gels, dispersible tablets, pills, capsules, powders, sustained release formulations or elixirs, for oral administration or in sterile solutions orsuspensions for parenteral administration, as well as transdermal patch preparation and dry powder inhalers. In one embodiment, the compounds described above are formulated into pharmaceutical compositions using techniques and procedures well known in the art (see, e.g., Ansel Introduction to Pharmaceutical Dosage Forms, Fourth Edition 1985, 126).

[0029] In the compositions, effective concentrations of one or more compounds or pharmaceutically acceptable derivatives thereof is (are) mixed with a suitable pharmaceutical carrier. The compounds may be derivatized as the corresponding salts, esters, acids, bases, solvates, hydrates or prodrugs prior to formulation, as described above. The concentrations of the compounds in the compositions are effective for delivery of an amount, upon administration, that treats, prevents, or ameliorates one or more of the symptoms of diseases or disorders to be treated.

[0030] In one embodiment, the compositions are formulated for single dosage administration. To formulate a composition, the weight fraction of compound is dissolved, suspended, dispersed or otherwise mixed in a selected carrier at an effective concentration such that the treated condition is relieved, prevented, or one or more symptoms are ameliorated.

[0031] The active compound is included in the pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the absence of undesirable side effects on the patient treated. The therapeutically effective concentration may be determined empirically by testing the compounds in in vitro and in vivo systems described herein, and then extrapolated from there for dosages for humans.

[0032] The concentration of active compound in the pharmaceutical composition will depend on absorption, distribution, inactivation and excretion rates of the active compound, the physicochemical characteristics of the compound, the dosage schedule, and amount administered as well as other factors known to those of skill in the art.

[0033] In one embodiment, a therapeutically effective dosage should produce a serum concentration of active ingredient of from about 0.1 ng / mL to about 50 - 100 µg / mL. The pharmaceutical compositions, in another embodiment, should provide a dosage of from about 0.001 mg to about 2000 mg of compound per kilogram of body weight per day. Pharmaceutical dosage unit forms are prepared to provide from about 0.01 mg, 0.1 mg or 1 mg to about 500 mg, 1000 mg or 2000 mg, and in one embodiment from about 10 mg to about 500 mg of the active ingredient or a combination of essential ingredients per dosage unit form.

[0034] Dosing may occur at intervals of minutes, hours, days, weeks, months or years or continuously over any one of these periods. Suitable dosages lie within the range of about 0.1 ngper kg of body weight to 1 g per kg of body weight per dosage. The dosage is preferably in the range of 1 µg to 1 g per kg of body weight per dosage, such as is in the range of 1 mg to 1 g per kg of body weight per dosage. Suitably, the dosage is in the range of 1 µg to 500 mg per kg of body weight per dosage, such as 1 µg to 200 mg per kg of body weight per dosage, or 1 µg to 100 mg per kg of body weight per dosage. Other suitable dosages may be in the range of 1 mg to 250 mg per kg of body weight, including 1 mg to 10, 20, 50 or 100 mg per kg of body weight per dosage or 10 µg to 100 mg per kg of body weight per dosage.

[0035] Suitable dosage amounts and dosing regimens can be determined by the attending physician and may depend on the particular condition being treated, the severity of the condition, as well as the general health, age and weight of the subject.

[0036] In instances in which the compounds exhibit insufficient solubility, methods for solubilizing compounds may be used. Such methods are known to those of skill in this art, and include, but are not limited to, using cosolvents, such as dimethyl sulfoxide (DMSO), using surfactants, such as TWEEN®, dissolution in aqueous sodium bicarbonate, formulating the compounds of interest as nanoparticles, and the like. Derivatives of the compounds, such as prodrugs of the compounds may also be used in formulating effective pharmaceutical compositions.

[0037] Upon mixing or addition of the compound(s), the resulting mixture may be a solution, suspension, emulsion or the like. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the disease, disorder or condition treated and may be empirically determined.

[0038] The pharmaceutical compositions are provided for administration to humans and animals in unit dosage forms, such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil-water emulsions containing suitable quantities of the compounds or pharmaceutically acceptable derivatives thereof. The pharmaceutically therapeutically active compounds and derivatives thereof are, in one embodiment, formulated and administered in unit-dosage forms or multiple-dosage forms. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at intervals of time. Unit-dose forms as used herein refers to physically discrete units suitable for human and animal subjects and packaged individually as is known in the art. Each unit-dose contains a predetermined quantity of the therapeutically active compound sufficient to produce the desired therapeutic effect, in association with the required pharmaceutical carrier, vehicle or diluent. Examples of unit-dose forms include ampules andsyringes and individually packaged tablets or capsules. Unit-dose forms may be administered in fractions or multiples thereof. A multiple-dose form is a plurality of identical unit-dosage forms packaged in a single container to be administered in segregated unit-dose form. Examples of multiple-dose forms include vials, bottles of tablets or capsules or bottles of pints or gallons. Hence, multiple dose form is a multiple of unit-doses which are not segregated in packaging.

[0039] Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15th Edition, 1975.

[0040] Dosage forms or compositions containing active ingredient in the range of 0.005% to 100% (wt.%) with the balance made up from non-toxic carrier may be prepared. Methods for preparation of these compositions are known to those skilled in the art. The contemplated compositions may contain 0.001%-100% (wt.%) active ingredient, in one embodiment 0.1-95% (wt.%), in another embodiment 75-85% (wt.%).

[0041] Pharmaceutical preparations for oral use can be obtained by combining the active agent, such as the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, with solid excipients and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients may include fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as maize starch, wheat starch, rice starch, potato starch, gelatine, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. Treatments

[0042] The compositions may be used to treat a disease or disorder such as diabetes, heart disease, cardiovascular disease, Parkinson's disease, malaria, cancer (including lung, prostrate, bladder, ovarian, cervical, pancreatic, breast, liver, esophageal, gastric, adenoma, and melanoma), digestive disorders, autoimmune diseases, chronic inflammation, stroke, obesity, headaches and migraines, and neurodegenerative disorders, such as disruptive mood dysregulation disorder, major depressive disorder, treatment-resistant depression, persistent depressive disorder, post-partum depression, an anxiety disorder, a substance-related disorder or post-traumatic stress disorder (PTSD).

[0043] In a third aspect of the invention there is provided the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, or thepharmaceutical composition according to the second aspect for use in treating inflammation in a subject in need thereof, optionally wherein the inflammation is treatable by modulation of prostaglandin E2 synthetase 1 (mPGES-1) and / or arachidonic acid 5-lipoxygenase (5-LO).

[0044] In a fourth aspect of the invention there is provided use of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, or the pharmaceutical composition according to the second aspect, for the manufacture of a medicament for treating inflammation in a subject in need thereof, optionally wherein the inflammation is treatable by modulation of prostaglandin E2 synthetase 1 (mPGES-1) and / or arachidonic acid 5-lipoxygenase (5-LO).

[0045] In a fifth aspect of the invention there is provided a method of treating inflammation in a subject in need thereof, said method comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, or the pharmaceutical composition according to the second aspect, to the subject, optionally wherein the inflammation is treatable by modulation of prostaglandin E2 synthetase 1 (mPGES-1) and / or arachidonic acid 5-lipoxygenase (5-LO).

[0046] In a sixth aspect of the invention there is provided the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, or the pharmaceutical composition according to the second aspect for use in treating cancer in a subject in need thereof, optionally wherein the cancer is treatable by modulation of tropomyosin receptor kinase B (TrkB).

[0047] In a seventh aspect of the invention there is provided use of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, or the pharmaceutical composition according to the second aspect, for the manufacture of a medicament for treating cancer in a subject in need thereof, optionally wherein the cancer is treatable by modulation of tropomyosin receptor kinase B (TrkB).

[0048] In an eighth aspect of the invention there is provided a method of treating cancer in a subject in need thereof, said method comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, or the pharmaceutical composition according to the second aspect, to the subject, optionally wherein the cancer is treatable by modulation of tropomyosin receptor kinase B (TrkB).

[0049] In certain embodiments of the sixth, seventh or eighth aspect, the cancer is selected from the group consisting of transitional cell carcinoma, bladder, brain, breast, Kaposi sarcoma, leukemia, lung, melanoma, ovarian, pancreatic, colon, and prostate cancer.

[0050] In one embodiment the compounds of the invention may be administered in the form of a “prodrug”. The phrase “prodrug” refers to a compound that, upon in vivo administration, is metabolized by one or more steps or processes or otherwise converted to the biologically, pharmaceutically or therapeutically active form of the compound. Prodrugs can be prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to a compound described herein. Modes of Administration

[0051] Convenient modes of administration include injection (subcutaneous, intravenous, intraperitoneal etc.), oral administration, inhalation, transdermal application, topical creams or gels or powders, ocular, optic and nasal dosage forms, vaginal or rectal administration. Depending on the route of administration, the formulation and / or compound may be coated with a material to protect the compound from the action of enzymes, acids and other natural conditions which may inactivate the therapeutic activity of the compound. The compound may also be administered parenterally or intraperitoneally. Compositions for oral administration

[0052] Oral pharmaceutical dosage forms are either solid, gel or liquid. The solid dosage forms are tablets, capsules, granules, and bulk powders. Types of oral tablets include compressed, chewable lozenges and tablets which may be enteric-coated, sugar-coated, sustained release or film-coated. Capsules may be hard or soft gelatin capsules, while granules and powders may be provided in non-effervescent or effervescent form with the combination of other ingredients known to those skilled in the art. Solid compositions for oral administration

[0053] In certain embodiments, the formulations are solid dosage forms, in one embodiment, capsules or tablets. The tablets, pills, capsules, troches and the like can contain one or more of the following ingredients, or compounds of a similar nature: a binder; a lubricant; a diluent; a glidant; a disintegrating agent; a coloring agent; a sweetening agent; a flavoring agent; a wetting agent; an emetic coating; and a film coating. Examples of binders include microcrystalline cellulose, gum tragacanth, glucose solution, acacia mucilage, gelatin solution, molasses, polvinylpyrrolidine, povidone, crospovidones, sucrose and starch paste. Lubricants include talc, starch, magnesium or calcium stearate, lycopodium and stearic acid. Diluents include, forexample, lactose, sucrose, starch, kaolin, salt, mannitol and dicalcium phosphate. Glidants include, but are not limited to, colloidal silicon dioxide. Disintegrating agents include crosscarmellose sodium, sodium starch glycolate, alginic acid, corn starch, potato starch, bentonite, methylcellulose, agar and carboxymethylcellulose. Coloring agents include, for example, any of the approved certified water soluble FD and C dyes, mixtures thereof; and water insoluble FD and C dyes suspended on alumina hydrate. Sweetening agents include sucrose, lactose, mannitol and artificial sweetening agents such as saccharin, and any number of spray dried flavors. Flavoring agents include natural flavors extracted from plants such as fruits and synthetic blends of compounds which produce a pleasant sensation, such as, but not limited to peppermint and methyl salicylate. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene lauryl ether. Emetic- coatings include fatty acids, fats, waxes, shellac, ammoniated shellac and cellulose acetate phthalates. Film coatings include hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000 and cellulose acetate phthalate.

[0054] The compound, or pharmaceutically acceptable derivative thereof, could be provided in a composition that protects it from the acidic environment of the stomach. For example, the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition may also be formulated in combination with an antacid or other such ingredient.

[0055] When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents. The compounds can also be administered as a component of an elixir, suspension, syrup, wafer, sprinkle, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.

[0056] The active materials can also be mixed with other active materials which do not impair the desired action, or with materials that supplement the desired action, such as antacids, H2 blockers, and diuretics. The active ingredient is a compound or pharmaceutically acceptable derivative thereof as described herein. Higher concentrations, up to about 98% by weight of the active ingredient may be included.

[0057] In certain embodiments, tablets and capsules formulations may be coated as known by those of skill in the art in order to modify or sustain dissolution of the active ingredient. Thus, forexample, they may be coated with a conventional enterically digestible coating, such as phenylsalicylate, waxes and cellulose acetate phthalate. Liquid compositions for oral administration

[0058] Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions and / or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Aqueous solutions include, for example, elixirs and syrups. Emulsions are either oil-in-water or water-in-oil.

[0059] Liquid pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, or otherwise mixing an active compound as defined above and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting agents, emulsifying agents, solubilizing agents, pH buffering agents and the like, for example, acetate, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, and other such agents.

[0060] Elixirs are clear, sweetened, hydroalcoholic preparations. Pharmaceutically acceptable carriers used in elixirs include solvents. Syrups are concentrated aqueous solutions of a sugar, for example, sucrose, and may contain a preservative. An emulsion is a two-phase system in which one liquid is dispersed in the form of small globules throughout another liquid. Pharmaceutically acceptable carriers used in emulsions are non-aqueous liquids, emulsifying agents and preservatives. Suspensions use pharmaceutically acceptable suspending agents and preservatives. Pharmaceutically acceptable substances used in non-effervescent granules, to be reconstituted into a liquid oral dosage form, include diluents, sweeteners and wetting agents. Pharmaceutically acceptable substances used in effervescent granules, to be reconstituted into a liquid oral dosage form, include organic acids and a source of carbon dioxide. Coloring and flavoring agents are used in all of the above dosage forms.

[0061] Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examples of preservatives include glycerin, methyl and propylparaben, benzoic acid, sodium benzoate and ethanol. Examples of non-aqueous liquids utilized in emulsions include mineral oil and cottonseed oil. Examples of emulsifying agents include gelatin, acacia, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitan monooleate. Suspending agents include sodium carboxymethylcellulose, pectin, tragacanth, Veegum and acacia. Sweetening agents include sucrose, syrups, glycerin and artificial sweetening agents such as saccharin. Wetting agentsinclude propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene lauryl ether. Organic acids include citric and tartaric acid. Sources of carbon dioxide include sodium bicarbonate and sodium carbonate. Coloring agents include any of the approved certified water soluble FD and C dyes, and mixtures thereof. Flavoring agents include natural flavors extracted from plants such fruits, and synthetic blends of compounds which produce a pleasant taste sensation.

[0062] For a solid dosage form, the solution or suspension, in for example propylene carbonate, vegetable oils or triglycerides, is in one embodiment encapsulated in a gelatin capsule. For a liquid dosage form, the solution, e.g., for example, in a polyethylene glycol, may be diluted with a sufficient quantity of a pharmaceutically acceptable liquid carrier, e.g., water, to be easily measured for administration.

[0063] Alternatively, liquid or semi-solid oral formulations may be prepared by dissolving or dispersing the active compound or salt in vegetable oils, glycols, triglycerides, propylene glycol esters (e.g., propylene carbonate) and other such carriers, and encapsulating these solutions or suspensions in hard or soft gelatin capsule shells. Other useful formulations include those set forth in U.S. Patent Nos. RE28,819 and 4,358,603. Briefly, such formulations include, but are not limited to, those containing a compound provided herein, a dialkylated mono- or poly- alkylene glycol, including, but not limited to, 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether wherein 350, 550 and 750 refer to the approximate average molecular weight of the polyethylene glycol, and one or more antioxidants, such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, thiodipropionic acid and its esters, and dithiocarbamates.

[0064] Other formulations include, but are not limited to, aqueous alcoholic solutions including a pharmaceutically acceptable acetal. Alcohols used in these formulations are any pharmaceutically acceptable water-miscible solvents having one or more hydroxyl groups, including, but not limited to, propylene glycol and ethanol. Acetals include, but are not limited to, di(lower alkyl) acetals of lower alkyl aldehydes such as acetaldehyde diethyl acetal. Injectables, Solutions and Emulsions

[0065] Parenteral administration, in one embodiment characterized by injection, either subcutaneously, intramuscularly, intraperitoneally or intravenously is also contemplated herein. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Theinjectables, solutions and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol or ethanol. In addition, if desired, the pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate and cyclodextrins.

[0066] Implantation of a slow-release or sustained-release system, such that a constant level of dosage is maintained is also contemplated herein. Briefly, a compound provided herein is dispersed in a solid inner matrix, e.g., polymethylmethacrylate, polybutylmethacrylate, plasticized or unplasticized polyvinylchloride, plasticized nylon, plasticized polyethyleneterephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonate copolymers, hydrophilic polymers such as hydrogels of esters of acrylic and methacrylic acid, collagen, cross-linked polyvinylalcohol and cross-linked partially hydrolyzed polyvinyl acetate, that is surrounded by an outer polymeric membrane, e.g., polyethylene, polypropylene, ethylene / propylene copolymers, ethylene / ethyl acrylate copolymers, ethylene / vinylacetate copolymers, silicone rubbers, polydimethyl siloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride, vinylchloride copolymers with vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubbers, ethylene / vinyl alcohol copolymer, ethylene / vinyl acetate / vinyl alcohol terpolymer, and ethylene / vinyloxyethanol copolymer, that is insoluble in body fluids. The compound diffuses through the outer polymeric membrane in a release rate controlling step. The percentage of active compound contained in such parenteral compositions is highly dependent on the specific nature thereof, as well as the activity of the compound and the needs of the subject.

[0067] Parenteral administration of the compositions includes intravenous, subcutaneous and intramuscular administrations. Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products, such as lyophilized powders, ready to be combined with a solvent just prior to use, including hypodermic tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with a vehicle just prior to use and sterile emulsions. The solutions may be either aqueous or nonaqueous.

[0068] If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol and mixtures thereof.

[0069] Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents and other pharmaceutically acceptable substances.

[0070] Examples of aqueous vehicles include Sodium Chloride Injection, Ringers Injection, Isotonic Dextrose Injection, Sterile Water Injection, Dextrose and Lactated Ringers Injection. Nonaqueous parenteral vehicles include fixed oils of vegetable origin, olive oil, cottonseed oil, corn oil, sesame oil and peanut oil. Antimicrobial agents in bacteriostatic or fungistatic concentrations must be added to parenteral preparations packaged in multiple-dose containers which include phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcelluose, hydroxypropyl methylcellulose and polyvinylpyrrolidone. Emulsifying agents include Polysorbate 80 (TWEEN^80). A sequestering or chelating agent of metal ions include ethylenediaminetetraacetic acid (EDTA). Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol and propylene glycol for water miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.

[0071] The concentration of the pharmaceutically active compound is adjusted so that an injection provides an effective amount to produce the desired pharmacological effect. The exact dose depends on the age, weight and condition of the patient or animal as is known in the art.

[0072] The unit-dose parenteral preparations are packaged in an ampule, a vial or a syringe with a needle. All preparations for parenteral administration must be sterile, as is known and practiced in the art.

[0073] Illustratively, intravenous or intraarterial infusion of a sterile aqueous solution containing an active compound is an effective mode of administration. Another embodiment is a sterile aqueous or oily solution or suspension containing an active material injected as necessary to produce the desired pharmacological effect.

[0074] Injectables are designed for local and systemic administration. In one embodiment, a therapeutically effective dosage is formulated to contain a concentration of at least about 0.1% w / w up to about 90% w / w or more, in certain embodiments more than 1% w / w of the active compound to the treated tissue(s).

[0075] The compound may be suspended in micronized or other suitable form or may be derivatized to produce a more soluble active product or to produce a prodrug. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the condition and may be empirically determined. Lyophilized Powders

[0076] Of interest herein are also lyophilized powders, which can be reconstituted for administration as solutions, emulsions and other mixtures. They may also be reconstituted and formulated as solids or gels.

[0077] The sterile, lyophilized powder is prepared by dissolving a compound provided herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent. The solvent may contain an excipient which improves the stability or other pharmacological component of the powder or reconstituted solution, prepared from the powder. Excipients that may be used include, but are not limited to, dextrose, sorbital, fructose, corn syrup, xylitol, glycerin, glucose, sucrose or other suitable agent. The solvent may also contain a buffer, such as citrate, sodium or potassium phosphate or other such buffer known to those of skill in the art at, in one embodiment, about neutral pH. Subsequent sterile filtration of the solution followed by lyophilization under standard conditions known to those of skill in the art provides the desired formulation. In one embodiment, the resulting solution will be apportioned into vials for lyophilization. Each vial will contain a single dosage or multiple dosages of the compound. The lyophilized powder can be stored under appropriate conditions, such as at about 4 °C to room temperature.

[0078] Reconstitution of this lyophilized powder with water for injection provides a formulation for use in parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or other suitable carrier. The precise amount depends upon the selected compound. Such amount can be empirically determined. Topical Administration

[0079] Topical mixtures are prepared as described for the local and systemic administration. The resulting mixture may be a solution, suspension, emulsions or the like and are formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, dermal patches or any other formulations suitable for topical administration. The compounds or pharmaceutically acceptablederivatives thereof may have increased lipid solubility as compared with cannflavin A and cannflavin B, in view of their longer carbon chain, and as such may be better suited for topical administration.

[0080] The compounds or pharmaceutically acceptable derivatives thereof may be formulated as aerosols for topical application, such as by inhalation. These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a microfine powder for insufflation, alone or in combination with an inert carrier such as lactose. In such a case, the particles of the formulation will, in one embodiment, have diameters of less than 50 microns, in one embodiment less than 10 microns.

[0081] The compounds may be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions and for application to the eye or for intracisternal or intraspinal application. Topical administration is contemplated for transdermal delivery and also for administration to the eyes or mucosa, or for inhalation therapies. Nasal solutions of the active compound alone or in combination with other pharmaceutically acceptable excipients can also be administered.

[0082] These solutions, particularly those intended for ophthalmic use, may be formulated as 0.01% - 10% (vol.%) isotonic solutions, pH about 5-7, with appropriate salts. Compositions for other routes of administration

[0083] Other routes of administration, such as transdermal patches, including iontophoretic and electrophoretic devices, vaginal and rectal administration, are also contemplated herein.

[0084] Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those of skill in the art. For example, pharmaceutical dosage forms for rectal administration are rectal suppositories, capsules and tablets for systemic effect. Rectal suppositories used herein mean solid bodies for insertion into the rectum which melt or soften at body temperature releasing one or more pharmacologically or therapeutically active ingredients. Pharmaceutically acceptable substances utilized in rectal suppositories are bases or vehicles and agents to raise the melting point. Examples of bases include cocoa butter (theobroma oil), glycerin-gelatin, carbowax (polyoxyethylene glycol) and appropriate mixtures of mono-, di- and triglycerides of fatty acids. Combinations of the various bases may be used. Agents to raise the melting point of suppositories include spermaceti and wax. Rectal suppositories may be prepared either by the compressed method or by molding. The weight of a rectal suppository, in one embodiment, is about 2 to 3 gm.

[0085] Tablets and capsules for rectal administration are manufactured using the same pharmaceutically acceptable substance and by the same methods as for formulations for oral administration. Targeted Formulations

[0086] The compounds provided herein, or pharmaceutically acceptable derivatives thereof, may also be formulated to be targeted to a particular tissue, receptor, or other area of the body of the subject to be treated. Many such targeting methods are well known to those of skill in the art. All such targeting methods are contemplated herein for use in the instant compositions.

[0087] In one embodiment, liposomal suspensions, including tissue-targeted liposomes, such as tumor-targeted liposomes, may also be suitable as pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art. For example, liposome formulations may be prepared as described in U.S. Patent No.4,522,811. Briefly, liposomes such as multilamellar vesicles (MLV's) may be formed by drying down egg phosphatidyl choline and brain phosphatidyl serine (7:3 molar ratio) on the inside of a flask. A solution of a compound provided herein in phosphate buffered saline lacking divalent cations (PBS) is added and the flask shaken until the lipid film is dispersed. The resulting vesicles are washed to remove unencapsulated compound, pelleted by centrifugation, and then resuspended in PBS. Co-administration with other drugs

[0088] In accordance with another aspect of the present invention, it is contemplated that compounds of Formula (I) as described herein may be administered to a subject in need thereof in combination with medication considered by those of skill in the art to be current standard of care for the condition of interest. Such combinations provide one or more advantages to the subject, e.g., requiring reduced dosages to achieve similar benefit, obtaining the desired palliative effect in less time, and the like.

[0089] Compounds in accordance with the present invention may be administered as part of a therapeutic regimen with other drugs. It may be desirable to administer a combination of active compounds, for example, for the purpose of treating a particular disease or condition. Accordingly, it is within the scope of the present invention that two or more pharmaceutical compositions, at least one of which contains a compound of Formula (I) according to the present invention, may be combined in the form of a kit suitable for co-administration of the compositions.

[0090] When two or more active ingredients are co-administered, the active ingredients may be administered simultaneously, sequentially or separately. In one embodiment the compound ofFormula (I) is co-administered simultaneously with a second therapeutic agent. In another embodiment the compound of Formula (I) and the second therapeutic agent are administered sequentially. In a further embodiment the compound of Formula (I) and the second therapeutic agent are administered separately.

[0091] Any of the features described herein can be combined in any combination with any one or more of the other features described herein within the scope of the invention. Method of producing the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof

[0092] In a ninth aspect of the invention there is provided a method of producing the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, said method comprising the following steps: preparing a reaction mixture comprising: chrysoeriol or apigenin, a poly-prenyl pyrophosphate, a prenyltransferase or a nucleic acid encoding a prenyltransferase; and incubating the reaction mixture under conditions suitable for the prenyltransferase to catalyse prenylation of the chrysoeriol or apigenin using the poly-prenyl pyrophosphate as a prenyl donor to thereby produce the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof.

[0093] The following options may be used in conjunction with the ninth aspect, either individually or in any combination.

[0094] In certain embodiments, the prenyltransferase catalyses prenylation of chrysoeriol or apigenin at the 6-position thereof.

[0095] In certain embodiments, the reaction mixture further comprises a cell lysate. In certain specific embodiments, the cell lysate is derived from cells engineered to express the prenyltransferase. In certain specific embodiments, the cell lysate is a cell-free protein expression lysate. In certain specific embodiments, the cell lysate is an E. coli cell lysate or a Leishmania cell lysate.

[0096] In certain embodiments, the reaction mixture comprises a nucleic acid encoding the prenyltransferase. In certain specific embodiments, the reaction mixture comprises RNA encoding the prenyltransferase, and the reaction mixture is incubated under conditions suitable for the RNA to be translated to produce the prenyltransferase. In certain specific embodiments,the reaction mixture comprises DNA encoding the prenyltransferase, and the reaction mixture is incubated under conditions suitable for the DNA to be transcribed to RNA and for the RNA to be translated to produce the prenyltransferase.

[0097] In certain embodiments, the prenyltransferase comprises the sequence set forth in SEQ ID NO: 1, or a sequence having at least about 80% identity to SEQ ID NO: 1. In certain specific embodiments, the prenyltransferase lacks a chloroplast transit peptide sequence. In certain specific embodiments, the chloroplast transit peptide sequence comprises the sequence set forth in SEQ ID NO: 3, or a sequence having at least about 80% identity to SEQ ID NO: 3. In certain specific embodiments, the prenyltransferase is between about 300 and 350 amino acids in length. In certain specific embodiments, the prenyltransferase comprises the sequence set forth in SEQ ID NO: 2, or a sequence having at least about 80% identity to SEQ ID NO: 2. In certain specific embodiments, the prenyltransferase comprises one or more mutations at a leader peptide cleavage site. In certain specific embodiments, the prenyltransferase is Cannabis sativa PT3 or a variant thereof. Sequences

[0098] Table 1 lists various enzymes and peptides that are relevant to the present disclosure. Table 1. Enzymes and peptides relevant to the present disclosure

[0099] Table 2 provides the sequences of the enzymes and peptides listed in Table 1.Table 2. Sequences relevant to the present disclosureCell-free expression [000100] The present disclosure provides cell-free reaction mixtures comprising a cell lysate, chemical substrates, and an enzyme or a nucleic acid encoding an enzyme. In examples where the cell-free reaction mixture comprises a nucleic acid encoding an enzyme, the cell lysate is preferably a cell-free protein expression lysate. A cell-free protein expression lysate is a cell lysate capable of supporting protein expression from a nucleic acid template, such as a DNA or RNA template. [000101] Cell-free protein expression is sometimes referred to as in vitro protein expression, in vitro translation, cell-free translation, cell-free expression and cell-free protein synthesis, and involves synthesising proteins in cell lysate rather than within cultured cells. Unlike in vivo techniques, cell-free expression does not require gene transfection, cell culture or extensive protein purification. [000102] Cell-free protein expression lysates typically provide the requisite enzymes and building blocks required for translation, such as ribosomes, amino acids, polymerases, nucleotides, tRNAs and other synthesis factors, although an energy source and amino acids may be added to the lysate in order to sustain protein synthesis. Cell membranes may be removed from the cell lysate, leaving the cytosolic and organelle components of the cell to carry outexpression. The cell lysate may be supplemented with components that enhance expression, such as amino acids, an energy source, ribonucleotide triphosphates, magnesium (e.g., magnesium chloride or magnesium acetate), spermidine, dithiothreitol, creatine phosphate, polyethylene glycol, a protease inhibitor, an oligonucleotide, creatine phosphokinase or an RNA polymerase (e.g., T7 RNA polymerase). [000103] Those skilled in the art will be aware of various cell-free protein expression systems that are commercially available. The cell-free protein expression systems of the present disclosure may be eukaryotic or prokaryotic. Cell-free expression lysates that may be suitable for use in the present disclosure include E. coli lysates, Leishmania lysates, wheat germ lysates, rabbit cell lysates, human cell lysates, insect cell lysates. Eukaryotic cell-free protein expression systems may be derived from insect cells, plant cells (e.g., wheat germ extracts), yeast cells or animal cells such as mammalian cells (e.g., rabbit cells or human cells). Prokaryotic cell-free protein expression systems may, for example, be derived from E. coli. Examples of E. coli cell- free expression systems are described in Garenne et al. Nature Reviews (2021)1:49. In some examples, the cell-free expression system is a Leishmania tarentolae cell-free expression system. Methods of preparing and performing Leishmania tarentolae cell-free expression systems are described in Kovtun et al. Methods (2011) 55: 58-64 and in Johnston, W.A., Moradi, S.V., Alexandrov, K. (2019). Adaption of the Leishmania Cell-Free Expression System to High- Throughput Analysis of Protein Interactions. In: Vincentelli, R. (eds) High-Throughput Protein Production and Purification. Methods in Molecular Biology, vol 2025. Humana, New York, NY. https: / / doi.org / 10.1007 / 978-1-4939-9624-7_19. The Leishmania tarentolae translation extract (LTE) system is a coupled transcription / translation system where DNA inputs are transcribed to mRNA via a heterologous RNA polymerase, and these mRNAs are subsequently translated by Leishmania ribosomes. Alternatively, translation can be initiated by direct addition of mRNA to the LTE system (Mureev et al. Nat Biotechnology (2009) 27, 747-752). [000104] Some cell lysates such as those derived from E. coli and wheat germ may have no or low amounts of endogenous mRNA. Other cell lysates may comprise endogenous mRNAs and may therefore be treated with a nuclease (e.g., micrococcal nuclease) to increase translation of an exogenously supplied mRNA. Once endogenous transcripts are removed, the lysate may be supplemented with an RNase inhibitor to prevent or reduce further mRNA degradation. [000105] Cell-free expression systems may support protein synthesis from DNA templates (transcription and translation) or mRNA templates (translation only). Cell-free expression systems may be designed to perform transcription and translation steps as two separate sequential reactions or concurrently as one reaction. Template DNA for cell-free transcriptionmay be linear, a circular plasmid or a PCR fragment. Preferably, the DNA to be transcribed is operably linked to a promoter sequence. Certain DNA-dependent RNA polymerases (RNA Pol) have only one subunit (e.g., those from bacteriophages such as T3 and T7, and mitochondria), while other RNA Pols from bacteria and eukaryotes are multi-subunit enzymes that require additional protein factors for efficient transcription. Monomeric RNA Pols from bacteriophages may be capable of performing transcription, including termination and release of the transcript from a DNA template, without the aid of additional protein factors. [000106] Transcripts for use in prokaryotic cell-free protein expression systems may require the ‘Shine-Dalgarno’ sequence to assist in proper translation. Eukaryotic cell-free transcription may include pre-mRNA processing to create a mature mRNA. This may include addition of a 5’ cap, addition of a Kozak sequence upstream of the initiation codon, addition of a poly-adenine tail and splicing of introns. These sequence elements may be incorporated into the DNA so that the immediate product of transcription is equivalent to a mature mRNA. [000107] An advantage of cell-free protein expression systems as compared to in vivo methods includes the ease with which different reaction conditions and parameters can be tested, including nucleic acid sequences, protein sequences and substrate concentrations. Indeed, the present inventors were able to test the activity of different enzymes and their variants, and in so doing, developed a prenyltransferase variant having advantageous activity relative to the full- length enzyme. [000108] Cell-free expression also enables the activity of an enzyme to be assayed without purifying the enzyme. Tagging or fusing the enzyme to another protein or peptide (e.g., a His tagging) is therefore typically not required. The enzymes of the present disclosure (e.g., methyltransferases, prenyltransferases and variants thereof) are preferably not fused or tagged to another protein or peptide sequence. In some examples, the enzymes of the present disclosure (e.g., methyltransferases, prenyltransferases and variants thereof) are not isolated or purified prior to incorporation to the cell-free reaction mixture. Prenylation of chrysoeriol or apigenin polyphenols [000109] The present disclosure provides methods of prenylating a polyphenol comprising: preparing a cell-free reaction mixture comprising a cell lysate, a polyphenol, a polyprenyl donor (e.g. a polyprenyl pyrophosphate),a prenyltransferase or a nucleic acid encoding a prenyltransferase; and incubating the cell-free reaction mixture under conditions suitable for the prenyltransferase to catalyse prenylation of the polyphenol using the polyprenyl donor as a substrate; wherein: the polyphenol is selected from the group consisting of chrysoeriol or apigenin; and the polyprenyl donor comprises three or more prenyl units. [000110] The present disclosure also provides systems for prenylating a polyphenol comprising: a cell lysate, optionally a cell-free protein expression lysate; a polyphenol; a polyprenyl donor (e.g. a polyprenyl pyrophosphate); and a prenyltransferase or a nucleic acid encoding a prenyltransferase; wherein: the prenyltransferase is capable of catalysing prenylation of the polyphenol using the polyprenyl donor as a substrate; the polyphenol is selected from the group consisting of chrysoeriol or apigenin; and the polyprenyl donor comprises three or more prenyl units. [000111] The present disclosure also provides methods of prenylating a polyphenol comprising: preparing a reaction mixture comprising cells engineered to express a prenyltransferase, a polyphenol, and a polyprenyl donor (e.g. a polyprenyl pyrophosphate); and incubating the reaction mixture under conditions suitable for the prenyltransferase to catalyse prenylation of the polyphenol using the polyprenyl donor as a substrate; wherein: the polyphenol is selected from the group consisting of chrysoeriol or apigenin; and the polyprenyl donor comprises three or more prenyl units. [000112] The present disclosure also provides systems for prenylating a polyphenol comprising: cells engineered to express a prenyltransferase,a polyphenol, and a polyprenyl donor (e.g. a polyprenyl pyrophosphate), wherein: the prenyltransferase is capable of catalysing prenylation of the polyphenol using the polyprenyl donor as a substrate; the polyphenol is selected from the group consisting of chrysoeriol or apigenin; and the polyprenyl donor comprises three or more prenyl units. [000113] In certain embodiments, the polyprenyl donor comprises three or more prenyl units. It may be, for example, tri-prenyl pyrophosphate (e.g. farnesyl diphosphate), tetra-prenyl pyrophosphate (e.g. geranylgeranyl diphosphate), penta-prenyl pyrophosphate, hexa-prenyl pyrophosphate, hepta-prenyl pyrophosphate, or octa-prenyl pyrophosphate. The polyprenyl donor may be farnesyl diphosphate or geranylgeranyl diphosphate. In some examples, the polyphenol is chrysoeriol. [000114] In some examples, the cell lysate is derived from a non-transgenic cell. In some examples, the cell lysate is derived from a cell that does not comprise a nucleic acid encoding the prenyltransferase. In some examples, the cell lysate is prepared by mixing the cell lysate, the chrysoeriol and the polyprenyl donor with the prenyltransferase or the nucleic acid encoding the prenyltransferase. In some examples, the cell lysate is derived from a cell that is not genetically modified. [000115] The cells of the present disclosure, or the cells from which cell lysate or the present disclosure is derived, may be eukaryotic cells, prokaryotic cells, microbial cells, bacterial cells, archaeon cells, protozoan cells, fungal cells, yeast cells, algae cells, plant cells, insect cells or animal cells. [000116] In some examples, the cell lysate is derived from cells engineered to express the prenyltransferase. Methods of producing the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof [000117] The present disclosure provides a method of producing the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof according to the first aspect, comprising: preparing a cell-free reaction mixture comprisinga cell lysate, chrysoeriol or apigenin, a poly-prenyl pyrophosphate, a prenyltransferase or a nucleic acid encoding a prenyltransferase; and incubating the cell-free reaction mixture under conditions suitable for the prenyltransferase to catalyse prenylation of the chrysoeriol or apigenin using the poly-prenyl pyrophosphate as a prenyl donor to produce the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof. [000118] In some examples, the reaction mixture is incubated at a temperature of between about 15 °C and 45 °C, such as between about 15 °C and 40 °C, or between about 20 °C and 40 °C, or between about 20 °C and 37 °C, or between about 20 °C and 35 °C, or between about 20 °C and 30 °C, or between about 22.5 °C and 30 °C, or between about 22.5 °C and 27.5 °C. In some examples, the reaction mixture is incubated at a temperature of about 20 °C, or about 21 °C, or about 22 °C, or about 23 °C, or about 24 °C, or about 25 °C, or about 26 °C, or about 27 °C, or about 28 °C, or about 29 °C, or about 30 °C, or about 31 °C, or about 32 °C, or about 33 °C, or about 34 °C, or about 35 °C or about 36 °C, or about 37 °C. In some examples, the reaction mixture is incubated at a temperature of about 25 °C. [000119] In some examples, the reaction mixture is incubated for between about 5 minutes and about 5 hours, such as between about 15 minutes and 5 hours, or between about 30 minutes and 5 hours, or between about 30 minutes and 4.5 hours, or between about 30 minutes and 4 hours, or between about 30 minutes and 3.5 hours, or between about 1 hour and 3.5 hours, or between about 1 hour and 3 hours, or between about 1.5 hours and 3 hours, or between about 2 hours and 3 hours. In some examples, the reaction mixture is incubated for about 1 hour, or about 1.5 hours, or about 2 hours, or about 2.5 hours, or about 3 hours. [000120] The cell-free reaction mixture may be supplemented with magnesium, optionally magnesium chloride. [000121] Preferably, the reaction mixture comprises a nucleic acid encoding the prenyltransferase. The nucleic acid is preferably DNA, and the reaction mixture is preferably incubated under conditions suitable for the DNA encoding the prenyltransferase to be transcribed to RNA and for the RNA encoding the prenyltransferase to be translated to produce the prenyltransferase. Expression of the nucleic acid (cell-free protein synthesis) may be carried out at a temperature of between about 15 °C and 40 °C, such as between about 15 °C and 40 °C, or between about 20 °C and 40 °C, or between about 20 °C and 37 °C, or between about 20 °C and35 °C, or between about 20 °C and 30 °C, or between about 22.5 °C and 30 °C, or between about 22.5 °C and 27.5 °C. In some examples, the cell-free protein synthesis is carried out at a temperature of about 20 °C, or about 21 °C, or about 22 °C, or about 23 °C, or about 24 °C, or about 25 °C, or about 26 °C, or about 27 °C, or about 28 °C, or about 29 °C, or about 30 °C, or about 31 °C, or about 32 °C, or about 33 °C, or about 34 °C, or about 35 °C or about 36 °C, or about 37 °C. In some examples, the cell-free protein synthesis is carried out at a temperature of about 25 °C. [000122] In some examples, the cell-free protein synthesis is carried out for between about 5 minutes and about 5 hours, such as between about 15 minutes and 5 hours, or between about 30 minutes and 5 hours, or between about 30 minutes and 4.5 hours, or between about 30 minutes and 4 hours, or between about 30 minutes and 3.5 hours, or between about 1 hour and 3.5 hours, or between about 1 hour and 3 hours, or between about 1.5 hours and 3 hours, or between about 2 hours and 3 hours. In some examples, the cell-free protein synthesis is carried out for about 1 hour, or about 1.5 hours, or about 2 hours, or about 2.5 hours, or about 3 hours. [000123] The chrysoeriol or apigenin, poly-prenyl pyrophosphate and optionally magnesium may be added after initiation of cell-free protein synthesis. In some examples, the chrysoeriol or apigenin, poly-prenyl pyrophosphate and optionally magnesium are added between about 10 minutes and 5 hours after initiation of cell-free protein synthesis, such as between about 15 minutes and 5 hours after initiation of cell-free protein synthesis, or between about 30 minutes and 5 hours, or between about 30 minutes and 4.5 hours, or between about 30 minutes and 4 hours, or between about 30 minutes and 3.5 hours, or between about 1 hour and 3.5 hours, or between about 1 hour and 3 hours, or between about 1.5 hours and 3 hours, or between about 2 hours and 3 hours after initiation of cell-free protein synthesis. In some examples, the chrysoeriol or apigenin, poly-prenyl pyrophosphate and optionally magnesium are added about 1 hour, or about 1.5 hours, or about 2 hours, or about 2.5 hours, or about 3 hours after initiation of cell-free protein synthesis. [000124] Following addition of the chrysoeriol or apigenin, poly-prenyl pyrophosphate and optionally magnesium, the reaction mixture may be incubated at a temperature of between about 20 °C and 45 °C, such as between about 25 °C and 45 °C, or between about 25 °C and 40 °C, or between about 27.5 °C and 40 °C, or between about 30 °C and 40 °C, or between about 32.5 °C and 40 °C, or between about 35 °C and 40 °C. In some examples, reaction mixture is incubated at a temperature of about 30 °C, or about 31 °C, or about 32 °C, or about 33 °C, or about 34 °C, or about 35 °C, or about 36 °C, or about 37 °C, or about 38 °C, or about 39 °C, or about 40 °C. In some examples, the reaction mixture is incubated at a temperature of about 37 °C.[000125] Following addition of the chrysoeriol or apigenin, poly-prenyl pyrophosphate and optionally magnesium, the reaction mixture may be incubated for about 10 minutes and 4 hours, such as between about 30 minutes and 4 hours, or between about 30 minutes and 3.5 hours, or between about 45 minutes and 3.5 hours, or between about 45 minutes and 3 hours, or between about 45 minutes and 2.5 hours, or between about 1 hour and 2.5 hours, or between about 1 hour and 2 hours. In some examples, the reaction mixture is incubated for about 30 minutes, or about 1 hour, or about 1.5 hours, or about 2 hours, or about 2.5 hours, or about 3 hours. In some examples, the reaction mixture is incubated for about 1.5 hours. Prenyltransferases [000126] The present disclosure provides prenyltransferases capable of catalysing prenylation of a polyphenol such as chrysoeriol or apigenin using a poly prenyl donor such as a poly-prenyl pyrophosphate as a substrate. The prenyltransferase may be a microbial prenyltransferase, a prokaryotic prenyltransferase, a eukaryotic prenyltransferase, an animal prenyltransferase, a plant prenyltransferase, a bacterial prenyltransferase, a protozoan prenyltransferase, a yeast prenyltransferase or an algae prenyltransferase. Various prenyltransferases and methods for identifying prenyltransferases are described in Rea et al. Phytochemistry (2019) 164:162-171 and in International Publication Nos. WO 2021 / 053513 A1 and WO 2022 / 109736 A1. [000127] Suitable prenyltransferases may include a Cannabis sativa prenyltransferase (CsPT), geranylgeranyl diphosphate (GGPP) synthase, hexaprenyl diphosphate (HexPP) synthase, heptaprenyl diphosphate (HepPP) synthase, octaprenyl (OPP) diphosphate synthase, solanesyl diphosphate (SPP) synthase, decaprenyl diphosphate (DPP) synthase, chicle synthase, and gutta- percha synthase; and a Z-isoprenyl diphosphate synthase, including, but not limited to, nonaprenyl diphosphate (NPP) synthase, undecaprenyl diphosphate (UPP) synthase, dehydrodolichyl diphosphate synthase, eicosaprenyl diphosphate synthase, natural rubber synthase, and other isoprenyl diphosphate synthases. [000128] Known prenyltransferase sequences include farnesyl pyrophosphate synthetase (HFPS) (GenBank Accession No. J05262, Homo sapiens); farnesyl diphosphate synthetase (FPP) (GenBank Accession No. J05091, Saccharomyces cerevisiae); isopentenyl diphosphate:dimethylallyl diphosphate isomerase J05090, Saccharomyces cerevisiae); farnesyl pyrophosphate synthetase 2 (FPS2) / FPP synthetase 2 / farnesyl diphosphate synthase 2 (GenBank Accession No. At4g17190, Arabidopsis thaliana); geranylgeranyl diphosphate synthase (GGPPS) (GenBank Accession No. AY371321, Ginkgo biloba); geranylgeranyl pyrophosphate synthase (GGPS1) / GGPP synthetase / farnesyltranstransferase (GenBank Accession No. At4g36810, Arabidopsis thaliana); farnesyl, geranylgeranyl, geranylfarnesyl,hexaprenyl, heptaprenyl diphosphate synthase (SeIF-HepPS) (GenBank Accession No. AB016095, Synechococcus elongatus). [000129] Known Cannabis prenyltransferase sequences and those from Cannabis relatives include CsPT1 (GenBank Accession No. PK28436), CsPT2 (GenBank Accession No. PK02092), CsPT3 (GenBank Accession No. PK17697), CsPT4 (GenBank Accession No. PK15523), CsPT5 (GenBank Accession No. PK11068), CsPT6 (GenBank Accession No. PK13891), CsPT7 (GenBank Accession No. PK29226), CsPT8 (GenBank Accession No. PK07278), H1PT1 (GenBank Accession No. AB543053), H1PT2 (GenBank Accession No. KM222442), AtVTE2-1 (GenBank Accession No. AAM10489), GmVTE2-1 (GenBank Accession No. ABB70126), TaVTE2-1 (GenBank Accession No. ABB70123), ZmVTE2-1 (GenBank Accession No. ABB70122), ApVTE2-1 (GenBank Accession No. ABB70124), CpVTE2-1 (GenBank Accession No. ABB70125), AtVTE2-2 (GenBank Accession No. ABB70127), GmVTE2-2 (GenBank Accession No. ABB70128), OsVTE2-2 (GenBank Accession No. XP_015646905), OsHGGT (GenBank Accession No. AAP43913), HvHGGT (GenBank Accession No. AAP43911), TaHGGT (GenBank Accession No. AAP43912), SfNSDT-1 (GenBank Accession No. BAG12671), GuA6DT (GenBank Accession No. KJ123716), LaPTl (GenBank Accession No. AER35706), SfiLDT (GenBank Accession No. BAK52290), SfG6DT (GenBank Accession No. BAK52291), GmG4DT (GenBank Accession No. BAH22520), AhR4DT-1 (GenBank Accession No. AQM74172), AhR3'DT-1 (GenBank Accession No. AQM74173), AhR3'DT-2 (GenBank Accession No. AQM74174), AhR3'DT-4 (GenBank Accession No. AQM74176), C1PT1 (GenBank Accession No. BAP27988), PcPT (GenBank Accession No. BAO31627), PsPTl (GenBank Accession No. AJW31563), PsPT2 (GenBank Accession No. AJW31564), MaIDT (GenBank Accession No. AJD80982) and CtIDT (GenBank Accession No. AJD80983). [000130] In some examples, the prenyltransferase is a plant prenyltransferase. In some examples, the prenyltransferase is a Cannabis prenyltransferase. In some examples, the prenyltransferase is a Cannabis sativa prenyltransferase. In some examples, the prenyltransferase is PT3 or a variant thereof, optionally Cannabis sativa PT3 or a variant thereof. [000131] In some examples, the prenyltransferase comprises the sequence set forth in SEQ ID NO: 1, or a sequence having at least about 70% identity to SEQ ID NO: 1. In some examples, the prenyltransferase has at least about 75% identity to SEQ ID NO: 1, or at least about 80% identity to SEQ ID NO: 1, or at least about 85% identity to SEQ ID NO: 1, or at least about 90% identity to SEQ ID NO: 1, or at least about 91% identity to SEQ ID NO: 1, or at least about 92% identity to SEQ ID NO: 1, or at least about 93% identity to SEQ ID NO: 1, or at least about 94% identityto SEQ ID NO: 1, or at least about 95% identity to SEQ ID NO: 1, or at least about 96% identity to SEQ ID NO: 1, or at least about 97% identity to SEQ ID NO: 1, or at least about 98% identity to SEQ ID NO: 1, or at least about 99% identity to SEQ ID NO: 1, or 100% identity to SEQ ID NO: 1. [000132] In some examples, the prenyltransferase lacks a plastid or chloroplast transit peptide sequence. Chloroplast transit peptides (CTPs) are typically cleaved by proteases after import into the chloroplast Emanuelsson et al. Protein Science (1999) 8: 978-984. Those skilled in the art will understand that a plastid or chloroplast transit peptide sequence may be identified using various techniques, including homology-based techniques and using annotated databases and scientific literature. TargetP 2.0 is a publicly available bioinformatic tool that may be used to identify plastid transit peptide sequences (Armenteros et al. Life Science Alliance (2019) 2). [000133] In some examples, the chloroplast transit peptide sequence comprises the sequence set forth in SEQ ID NO: 3, or a sequence having at least about 70% identity to SEQ ID NO: 3, such as at least about 75% identity to SEQ ID NO: 3, or at least about 80% identity to SEQ ID NO: 3, or at least about 85% identity to SEQ ID NO: 3, or at least about 90% identity to SEQ ID NO: 3, or at least about 91% identity to SEQ ID NO: 3, or at least about 92% identity to SEQ ID NO: 3, or at least about 93% identity to SEQ ID NO: 3, or at least about 94% identity to SEQ ID NO: 3, or at least about 95% identity to SEQ ID NO: 3, or at least about 96% identity to SEQ ID NO: 3, or at least about 97% identity to SEQ ID NO: 3, or at least about 98% identity to SEQ ID NO: 3, or at least about 99% identity to SEQ ID NO: 3, or 100% identity to SEQ ID NO: 3. [000134] In some examples, the prenyltransferase lacking a plastid peptide sequence is between about 250 and 450 amino acids in length, such as between about 275 and 450 amino acids in length, or between about 275 and 425 amino acids in length, or between about 275 and 410 amino acids in length, or between about 285 and 410 amino acids in length, or between about 285 and 400 amino acids in length, or between about 295 and 390 amino acids in length, or between about 300 and 390 amino acids in length, or between about 300 and 380 amino acids in length, or between about 300 and 370 amino acids in length, or between about 300 and 360 amino acids in length, or between about 300 and 350 amino acids in length, or between about 300 and 340 amino acids in length, or between about 300 and 330 amino acids in length, or between about 300 and 320 amino acids in length, or between about 305 and 320 amino acids in length, or between about 310 and 320 amino acids in length, or between about 315 and 320 amino acids in length, such as about 315, about 316, about 317, about 318, about 319 or about 320 amino acids in length.[000135] In some examples, the prenyltransferase lacking a plastid peptide sequence is less than about 400 amino acids in length, such as less than about 395 amino acids in length, or less than about 390 amino acids in length, or less than about 385 amino acids in length, or less than about 380 amino acids in length, or less than about 375 amino acids in length, or less than about 370 amino acids in length, or less than about 365 amino acids in length, or less than about 360 amino acids in length, or less than about 355 amino acids in length, or less than about 350 amino acids in length, or less than about 345 amino acids in length, or less than about 340 amino acids in length, or less than about 335 amino acids in length, or less than about 330 amino acids in length, or less than about 325 amino acids in length, or less than about 320 amino acids in length, or less than about 315 amino acids in length, or less than about 310 amino acids in length, or less than about 305 amino acids in length, or less than about 300 amino acids in length. [000136] In some examples, the present disclosure provides an isolated or recombinant prenyltransferase capable of catalysing prenylation of a polyphenol such as chrysoeriol or apigenin, wherein the prenyltransferase lacks a chloroplast transit peptide sequence. In some examples, the polyphenol is chrysoeriol or apigenin. [000137] In some examples, the present disclosure provides an isolated or recombinant prenyltransferase capable of catalysing prenylation of chrysoeriol or apigenin, wherein the prenyltransferase lacks a chloroplast transit peptide sequence. In some examples, the prenyltransferase catalyses prenylation of the chrysoeriol or apigenin using a polyprenyl donor. In certain examples, the poly prenyl donor is farnesyl diphosphate (FPP), and / or geranylgeranyl diphosphate. [000138] In some examples, the prenyltransferase lacking the chloroplast transit peptide sequence comprises the sequence set forth in SEQ ID NO: 2, or a sequence having at least about 70% identity to SEQ ID NO: 2, or at least about 75% identity to SEQ ID NO: 2, or at least about 80% identity to SEQ ID NO: 2, or at least about 85% identity to SEQ ID NO: 2, or at least about 90% identity to SEQ ID NO: 2, or at least about 91% identity to SEQ ID NO: 2, or at least about 92% identity to SEQ ID NO: 2, or at least about 93% identity to SEQ ID NO: 2, or at least about 94% identity to SEQ ID NO: 2, or at least about 95% identity to SEQ ID NO: 2, or at least about 96% identity to SEQ ID NO: 2, or at least about 97% identity to SEQ ID NO: 2, or at least about 98% identity to SEQ ID NO: 2, or at least about 99% identity to SEQ ID NO: 2, or 100% identity to SEQ ID NO: 2. [000139] In some examples, the prenyltransferase: comprises the sequence set forth in SEQ ID NO: 2, or a sequence having at least about 70% identity to SEQ ID NO: 2, or at least about 75% identity to SEQ ID NO: 2, or at least about80% identity to SEQ ID NO: 2, or at least about 85% identity to SEQ ID NO: 2, or at least about 90% identity to SEQ ID NO: 2, or at least about 91% identity to SEQ ID NO: 2, or at least about 92% identity to SEQ ID NO: 2, or at least about 93% identity to SEQ ID NO: 2, or at least about 94% identity to SEQ ID NO: 2, or at least about 95% identity to SEQ ID NO: 2, or at least about 96% identity to SEQ ID NO: 2, or at least about 97% identity to SEQ ID NO: 2, or at least about 98% identity to SEQ ID NO: 2, or at least about 99% identity to SEQ ID NO: 2, or 100% identity to SEQ ID NO: 2; and lacks a chloroplast transit peptide sequence comprising the sequence set forth in SEQ ID NO: 3, or a sequence having at least about 70% identity to SEQ ID NO: 3, such as at least about 75% identity to SEQ ID NO: 3, or at least about 80% identity to SEQ ID NO: 3, or at least about 85% identity to SEQ ID NO: 3, or at least about 90% identity to SEQ ID NO: 3, or at least about 91% identity to SEQ ID NO: 3, or at least about 92% identity to SEQ ID NO: 3, or at least about 93% identity to SEQ ID NO: 3, or at least about 94% identity to SEQ ID NO: 3, or at least about 95% identity to SEQ ID NO: 3, or at least about 96% identity to SEQ ID NO: 3, or at least about 97% identity to SEQ ID NO: 3, or at least about 98% identity to SEQ ID NO: 3, or at least about 99% identity to SEQ ID NO: 3, or 100% identity to SEQ ID NO: 3. [000140] In some examples, the prenyltransferase comprises one or more mutations at a peptide cleavage site, such as a leader peptide cleavage site. Those skilled in the art will understand that peptide cleavage sites can be identified in several ways, such as experimental determination of peptide cleavage sites by extraction and analysis of the target protein from native hosts or suitable heterologous protein expression systems, or bioinformatic prediction of peptide cleavage sites based on conserved amino acid sequence motifs and physicochemical amino acid properties associated with experimentally determined peptide cleavage sites. Common motifs marking the cleavage site of chloroplast transit peptides are described in Gavel and von Heijne FEBS (1990) 261(2), 455-458. TargetP2.0 (DOI: 0.26508 / lsa.201900429) is a bioinformatic tool that uses machine learning to assign probabilities to identify potential signal peptides cleavage sites. [000141] The prenyltransferase lacking the chloroplast transit peptide sequence may be used in a method of catalysing the prenylation of a polyphenol, such as chrysoeriol or apigenin. For example, the prenyltransferase may be recombinantly expressed, purified and used in a method of catalysing the prenylation of a polyphenol (e.g. chrysoeriol or apigenin). For example, the prenyltransferase may be expressed in a microorganism such as a yeast or a bacterium. Methods of recombinant protein expression and purification are well known in the art and are describedextensively in the literature such as in D. Amberg, D. Burke and J. Strathem, Methods in Yeast Genetics, 2005 Edition, Cold Spring Harbor Laboratory Press. [000142] The present disclosure also provides a nucleic acid encoding an enzyme described herein (e.g., a prenyltransferase or a variant thereof). In one example, the present disclosure provides an isolated nucleic acid encoding a prenyltransferase that lacks a plastid or chloroplast transit peptide sequence as described herein. The nucleic acid may be codon optimised for expression in a particular organism or by the cell-free translation machinery of a particular organism. The nucleic acid may be operably connected to a promoter. The promoter may be derived from a plant, an animal, a prokaryote, a protozoan, an algae, a fungus or a virus. In some examples, the promoter is not a plant promoter. In some examples, the promoter is not a Cannabis promoter. In some examples, the promoter is not a Cannabis sativa promoter. [000143] The present disclosure also provides a vector, such as a plasmid or viral vector, comprising a nucleic acid described herein. The present disclosure also provides a host cell comprising a nucleic acid or a vector described herein. In some examples, the host cell is capable of recombinantly expressing the enzyme encoded by the nucleic acid. BRIEF DESCRIPTION OF THE DRAWINGS [000144] The patent or application file contains at least one drawing executed in colour. Copies of this patent or patent application publication with colour drawings will be provided by the Office upon request and payment of the necessary fee (if appropriate). [000145] Figure 1: Chemical structures of (A) cannflavin A, (B) cannflavin B, and (C) cannflavin C. [000146] Figure 2: Compartmentalisation of prenyl diphosphate synthesis in plants. Plant cells have independent metabolic pathways for synthesis of isoprenoids in plastids (the methylerythritol phosphate pathway) and the cytosol (the mevalonate pathway). Both pathways produce the universal isoprenoid precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Distinct sets of enzymes metabolise downstream isoprenoid synthesis reactions in each of these compartments. Geranyl diphosphate (GPP) and geranylgeranyl diphosphate (GGPP) are only observed in the plastid, while farnesyl diphosphate (FPP) is only observed in the cytosol. [000147] Figure 3: Novel cannflavin molecules. (A) CsPT3 prenyltransferase reactions yielding products that are also observed in Cannabis sativa. (B) Products of CsPT3 prenyltransferase reactions, not observed previously in nature or in studies of the CsPT3 prenyltransferase enzyme.[000148] Figure 4: Biocatalytic synthesis of novel cannflavin molecules. Chrysoeriol was incubated with the CsPT3_trunc enzyme and GPP (A), DMAPP (B), FPP (C), or GGPP (D). Reaction products were separated via LC-MS / MS, monitoring for chrysoeriol (m / z = 301), cannflavin A (m / z = 437), cannflavin B (m / z = 369), or the predicted masses of Cfl15 (m / z = 505) or Cfl20 (m / z = 573). Known structures of cannflavin A and cannflavin B are shown, as well as predicted structures of Cfl15 and Cfl20. [000149] Figure 5: Cannflavin-15 product formation with trans,trans-farnesyl diphosphate. Chrysoeriol (200 μM) was incubated with CsPT3_trunc and different concentrations of trans,trans-FPP (squares, 25 μM; circles, 100 μM; triangles, 150 μM), or undetermined mixed isomers of FPP (crosses, 25 μM). Product formation was identified via HPLC. n = 2 replicate incubations per condition, each replicate plotted individually. [000150] Figure 6: Preparative HPLC purification of cannflavin-15. (A) A 10 μL volume of reaction product indicated a major peak with comparable retention time to chrysoeriol (2-2.5 min), and an additional minor peak between 11.5-12 min. The minor peak fraction between 11.5- 12 min was collected. (B) The purity of the collected fraction was verified by injection of a 10 μL sample. [000151] Figure 7:1H-NMR spectrum (acetone-d6) of cannflavin-15. [000152] Figure 8: Aromatic region of1H-NMR spectrum (acetone-d6) of cannflavin-15. [000153] Figure 9: Aliphatic region of1H-NMR spectrum (acetone-d6) of cannflavin-15. [000154] Figure 10: DEPTQ13513C-NMR spectrum (acetone-d6) of cannflavin-15. [000155] Figure 11: HMBC correlations for cannflavin-15. [000156] Figure 12: NOESY correlations for cannflavin-15. Dashed arrow designates chemical exchange. [000157] Figure 13: Synthesis of cannflavin-15 using E. coli lysates and Leishmania-based cell free protein synthesis. (A) LC-MS trace for an experiment in which CsPT3_trunc was expressed in E. coli BL21(DE3) and cell lysates were incubated with chrysoeriol (0.2 mM) and farnesyl diphosphate (0.2 mM). (B) LC-MS trace for a negative control experiment based on (A) but excluding plasmid DNA. (C) LC-MS trace for an experiment in which CsPT3_trunc was expressed in the Leishmania cell free protein expression system and incubated with chrysoeriol, farnesyl diphosphate and magnesium. (D) LC-MS trace for a negative control experiment based on (C) but excluding plasmid DNA. [000158] Figure 14: Cannflavin B standard curve.[000159] Figure 15: Schematic plasmid maps. Transcription in the LTE cell free expression system is regulated by the T7 promoter (indicated by a curved arrow). Transcripts include an unstructured 5’ untranslated region (5’ UTR) for recruitment of Leishmania ribosomes. The start codon is indicated (ATG) and all coding regions begin with a species-independent translational leader sequence (SITS) which is in frame with the subsequent gene of interest. Plasmids (A) pCF / CsPT3, and (B) pCF / CsPT3_trunc represent cell free expression plasmids for the luteolin O-methyltransferase, chrysoeriol prenyltransferase, and truncated chrysoeriol prenyltransferase, respectively. The predicted chloroplast transit peptide (CTP) in pCF / CsPT3 (A) is indicated with grey shading. Coding sequences are followed by a T7 terminator sequence. [000160] Figure 16: All cannflavin docking poses (light blue, including cannflavin A, cannflavin B, cannflavin 15 and cannflavin 20) for prostaglandin E synthase 2, showing consistent positioning above the reactive heme group (tan). DEFINITIONS [000161] In describing and claiming the present invention, the following terminology will be used in accordance with the definitions set out below. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only and is not intended to be limiting. [000162] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one having ordinary skill in the art to which the invention pertains. [000163] Unless the context clearly requires otherwise, throughout the description and the claims, the terms “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”. For example, a composition, mixture, process or method that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, mixture, process or method. [000164] The transitional phrase “consisting of” excludes any element, step, or ingredient not specified. If in the claim, such would close the claim to the inclusion of materials other than those recited except for impurities ordinarily associated therewith. When the phrase “consisting of” appears in a clause of the body of a claim, rather than immediately following the preamble, it limits only the element set forth in that clause; other elements are not excluded from the claim as a whole.[000165] The transitional phrase “consisting essentially of” is used to define a composition, process or method that includes materials, steps, features, components, or elements, in addition to those literally disclosed, provided that these additional materials, steps, features, components, or elements do not materially affect the basic and novel characteristic(s) of the claimed invention. The term “consisting essentially of” occupies a middle ground between “comprising” and “consisting of”. [000166] Where applicants have defined an invention or a portion thereof with an open-ended term such as “comprising”, it should be readily understood that (unless otherwise stated) the description should be interpreted to also describe such an invention using the terms “consisting essentially of” or “consisting of”. Thus, in some embodiments not otherwise explicitly recited, any instance of “comprising” may be replaced by “consisting of” or, alternatively, by “consisting essentially of”. [000167] Other than in the claims or operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein are to be understood as modified in all instances by the term “about”. The examples are not intended to limit the scope of the invention. [000168] In what follows, or where otherwise indicated, “%” will mean “weight %”, “ratio” will mean “weight ratio” and “parts” will mean “weight parts”. [000169] The terms “predominantly”, “predominant”, and “substantially” as used herein shall mean comprising more than 50% by weight, unless otherwise indicated. [000170] As used herein, with reference to numbers in a range of numerals, the terms “about,” “approximately” and “substantially” are understood to refer to the range of -10% to +10% of the referenced number, preferably -5% to +5% of the referenced number, more preferably -1 % to + 1 % of the referenced number, most preferably -0.1 % to +0.1 % of the referenced number. Moreover, with reference to numerical ranges, these terms should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, from 8 to 10, and so forth. [000171] The terms “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the invention.[000172] The term "identity" refers to a relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by aligning and comparing the sequences. The percent identity between two sequences is a function of the number of identical positions shared by the sequences when the sequences are optimally aligned (i.e., % homology = # of identical positions / total # of positions x 100), with optimal alignment determined taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. [000173] The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package, using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide or amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4: 11-17 (1989)) which has been incorporated into the ALIGN program, using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. In addition, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package, using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. Other computer programs that can be used in this regard include BLASTP, BLASTN and FASTA (Altschul et al., J. Molec. Biol., 1990:215:403). Another method for determining the percentage identity between two polypeptides involves the Clustal W algorithm (Thompson, J D, Higgines, D G and Gibson T J, 1994, Nucleic Acid Res 22(22): 4673-4680 together with the BLOSUM 62 scoring matrix (Henikoff S & Henikoff, JG, 1992, Proc. Natl. Acad. Sci. USA 89: 10915-10919) using a gap opening penalty of 10 and a gap extension penalty of 0.1, so that the highest order match obtained between two sequences wherein at least 50% of the total length of one of the two sequences is involved in the alignment. [000174] The term “isolated” as used herein refers to material that is substantially or essentially free from components that normally accompany it in its native state. For example, an isolated polynucleotide as used herein refers to a polynucleotide which has been purified from the sequences which flank it in a naturally occurring state, e.g., a DNA fragment which has been removed from the sequences that are normally adjacent to the fragment. Alternatively, an isolated enzyme, as used herein, may refer to in vitro isolation and / or purification of the enzyme from its cellular environment, and from association with other components of the cell, i.e., it isnot associated with in vivo substances. An isolated enzyme will generally encompass recombinantly expressed enzymes. [000175] As used herein, the term "operably connected" and its grammatical equivalents refers to the association of nucleic acid sequences on a single polynucleic acid fragment so that the function of one is affected by the other. For example, a promoter is operably linked with a coding sequence when it is capable of effecting the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter). [000176] The term "pharmaceutically acceptable" as used herein refers to substances that do not cause substantial adverse allergic or immunological reactions when administered to a subject. A "pharmaceutically acceptable carrier" includes, but is not limited to, solvents, coatings, dispersion agents, wetting agents, isotonic and absorption delaying agents and disintegrants. [000177] The term “polynucleotide variant” refers to polynucleotides displaying substantial sequence identity with a reference polynucleotide sequence or polynucleotides that hybridise with a reference sequence under stringent conditions. The term also encompasses polynucleotides that are distinguished from a reference polynucleotide by the addition, deletion or substitution of at least one nucleotide. Accordingly, the term “polynucleotide variant” includes polynucleotides in which one or more nucleotides have been added or deleted, or replaced with different nucleotides. In this regard, it is well understood in the art that certain alterations inclusive of mutations, additions, deletions and substitutions can be made to a reference polynucleotide whereby the altered polynucleotide retains the biological function or activity of the reference polynucleotide. The term “polynucleotide variant” also includes naturally occurring allelic variants. The terms “peptide variant”, “polypeptide variant”, “enzyme variant” and the like includes peptides, polypeptides and enzymes that are distinguished from a reference peptide, polypeptide or enzyme by the addition, deletion or substitution of at least one amino acid residue. In certain embodiments, a peptide, polypeptide or enzyme variant is distinguished from a reference peptide, polypeptide or enzyme by one or more substitutions, which may be conservative or non-conservative. In certain examples, the peptide, polypeptide or enzyme variant comprises conservative substitutions and, in this regard, it is well understood in the art that some amino acids may be changed to others with broadly similar properties without changing the nature of the activity of the peptide, polypeptide or enzyme. Peptide, polypeptide and enzyme variants also encompass peptides, polypeptides and enzymes in which one or more amino acids have been added or deleted, or replaced with different amino acid residues. [000178] As used herein, the term "recombinant" and its grammatical equivalents means that a particular nucleic acid (DNA or RNA) is the product of various combinations of cloning,restriction, and / or ligation steps resulting in a construct having a structural coding or non-coding sequence distinguishable from endogenous nucleic acids found in natural systems. Thus, as used herein, the terms "recombinant polynucleotide" or "recombinant nucleic acid" refer to a polynucleotide that is non-naturally occurring, e.g., is made by the artificial combination of two otherwise separated segments of sequence through human intervention. This artificial combination may be accomplished by chemical synthesis means, or by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques. [000179] The term "substantially complementary" when used to describe a first nucleotide sequence in relation to a second nucleotide sequence, refers to the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize to, and form a duplex structure with, an oligonucleotide or polynucleotide comprising the second nucleotide sequence. It will be understood that the sequence of a nucleic acid need not be 100% complementary to that of its target. Conditions under which hybridisation occurs may be stringent, such as 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 50oC or 70oC for 12-16 hours followed by washing. Other conditions, such as physiologically relevant conditions as may be encountered inside an organism, can also apply. Substantial complementarity allows the relevant function of the nucleic acid to proceed, e.g., guide RNA hybridisation and CRISPR-mediated gene activation. The skilled person will be able to determine the set of conditions most appropriate for a test of complementarity of two sequences in accordance with the ultimate application of the hybridized nucleotides. [000180] As used herein, the term "vector" and its grammatical equivalents refers to any means by which a nucleic acid can be propagated and / or transferred between organisms, cells, or cellular components. Vectors include viruses, bacteriophage, pro-viruses, plasmids, phagemids, transposons, and artificial chromosomes such as YACs (yeast artificial chromosomes), BACs (bacterial artificial chromosomes), and PLACs (plant artificial chromosomes), and the like, that are "episomes", that is, that replicate autonomously or can integrate into a chromosome of a host microorganism. A vector can also be a naked RNA polynucleotide, a naked DNA polynucleotide, a polynucleotide composed of both DNA and RNA within the same strand, a poly-lysine-conjugated DNA or RNA, a peptide-conjugated DNA or RNA, a liposome- conjugated DNA, or the like, that are not episomal in nature, or it can be an organism which comprises one or more polynucleotide constructs such as an agrobacterium or a bacterium. [000181] As used herein, the terms “tri-prenyl”, “tetra-prenyl”, “penta-prenyl”, “hexa-prenyl”, “hepta-prenyl”, and “octa-prenyl” mean a group containing three, four, five, six, seven, or eight prenyl units, respectively. In certain embodiments, the terms “tri-prenyl”, “tetra-prenyl”, “penta-prenyl”, “hexa-prenyl”, “hepta-prenyl”, and “octa-prenyl” mean groups having the following respective structures:[000182] As used herein, the term “poly-prenyl pyrophosphate” means a pyrophosphate containing more than one prenyl unit. For example, farnesyl diphosphate, being a tri-prenyl pyrophosphate, is an example of a poly-prenyl pyrophosphate. In certain embodiments, the term “poly-prenyl pyrophosphate” means a pyrophosphate containing three or more prenyl units. ABBREVIATIONS [000183] Adenosine triphosphate (ATP); Cannabis sativa prenyltransferase (CsPT); cannflavin- 15 (Cfl15); cannflavin-20 (Cfl20); chloroplast transit peptide (CTP); correlation spectroscopy (COSY); cytidine triphosphate (CTP); decaprenyl diphosphate (DPP); dimethylallyl diphosphate (DMAPP); ethylenediaminetetraacetic acid (EDTA); farnesyl diphosphate (FPP); geranylgeranyl diphosphate (GGPP); geranylgeranyl diphosphate synthase (GGPPS); geranyl diphosphate (GPP); guanosine triphosphate (GTP); heptaprenyl diphosphate (HepPP); heteronuclear multiple bond correlation (HMBC); heteronuclear single quantum correlation (HSQC): hexaprenyl diphosphate (HexPP); high performance liquid chromatography (HPLC); 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES); isopentenyl diphosphate (IPP); Leishmania tarentolae cell-free protein expression (LTE); liquid chromatography-mass spectrometry / mass spectrometry (LC-MS / MS); lysogeny broth (LB); nonaprenyl diphosphate (NPP); nuclear magnetic resonance (NMR); nuclear Overhauser effect spectroscopy (NOESY);octaprenyl (OPP); polyethylene glycol (PEG); species-independent translational leader sequence (SITS); solanesyl diphosphate (SPP); total correlation spectroscopy (TOCSY); tris(hydroxymethyl)aminomethane (Tris); undecaprenyl diphosphate (UPP); uridine triphosphate (UTP). [000184] Preferred features, embodiments and variations of the invention may be discerned from the following Examples which provide sufficient information for those skilled in the art to perform the invention. The following Examples are not to be regarded as limiting the scope of the preceding Summary of the Invention in any way. EXAMPLES Materials and methods Chemicals [000185] Analytical standards for chrysoeriol (Cat# C432820) and cannflavin B (Cat# C175405) were purchased from Toronto Research Chemicals (trc-canada.com). Dimethylallyl diphosphate (DMAPP, Cat# D4287), geranyl diphosphate (GPP, Cat# G6772), farnesyl diphosphate (FPP, Cat# F6892), trans,trans-farnesyl diphosphate (Cat# 44722), and geranylgeranyl diphosphate (GGPP, Cat# G6025) were purchased from Merck (Australia). Cannflavin A (Cat# FC171893) was purchased from Biosynth (biosynth.com). All other chemicals were the highest quality available from commercial suppliers. DNA constructs [000186] Nucleotide sequences for prenyltransferase (CsPT3) were optimised for Escherichia coli codon usage using the Integrated DNA Technologies (idtdna.com) codon optimization tool. Optimised sequences were synthesised by Twist Bioscience and individually cloned into the pET29b expression vector. The putative chloroplast transit peptide of CsPT3 was identified using TargetP 2.0 and a truncated variant (CsPT3_trunc) was also synthesised (Armenteros et al. Life Science Alliance (2019) 2). [000187] Plasmids for LTE-based cell-free expression were constructed by individually cloning the CsPT3, and CsPT3_trunc coding sequences into the pCellFree expression vector (Figure 15). Cloning into the pCellFree expression format adds an in-frame leader sequence to the 5’ end of the coding sequence to facilitate cell free protein synthesis with the LTE system. Cell-free protein synthesis [000188] The CsPT3 and CSPT3_trunc genes were expressed in LTE individually or in a pair depending on the designed enzymatic reactions. The detailed procedure for preparation of theLTE lysate and feeding solution were described in Kovtun et al. Methods (2011) 55, 58-64 and Adaption of the Leishmania Cell-Free Expression System to High-Throughput Analysis of Protein Interactions | SpringerLink. https: / / link.springer.com / protocol / 10.1007 / 978-1-4939-9624- 7_19. Briefly, a culture of actively dividing Leishmania tarentolae cells was harvested by centrifugation and washed twice in Sucrose Elution Buffer (SEB: 45 mM HEPES (pH 7.6), 250 mM sucrose, 100 mM potassium acetate, and 3 mM magnesium acetate). Washed cells were resuspended in SEB and disrupted using a nitrogen cavitation device (70 bar N2, 45 min equilibration at 4 °C). The lysate was centrifuged at 30000 g and sucrose was removed via gel filtration on a PD-10 Superdex 25 column (GE Healthcare) and eluted in fresh elution buffer (EB: HEPES (pH 7.6), 250 mM sucrose, 100 mM potassium acetate, and 3 mM magnesium acetate). The lysate was then snap-frozen and stored at -80 °C. Prior to protein expression the LTE extract was supplemented with the following feeding solution: 8 mM HEPES-KOH (pH 7.6), ribonucleotide triphosphates (1.7 mM ATP, 0.635 mM GTP, 0.5 mM CTP, 0.5 mM UTP), 3.35 mM magnesium acetate, 0.24 mM spermidine, 2 mM dithiothreitol, 40 mM creatine phosphate, 1% (v / v) PEG 3350, 1X complete cocktail protease inhibitor (Roche), 0.133 mM each proteinogenic amino acid, 0.2 mM anti-splice leader DNA oligonucleotide, 1 U / μL creatine phosphokinase, and 2 μg / mL T7 RNA polymerase. [000189] Protein expression reactions were performed in 150 μL volume containing DNA templates (70 ng / μL), LTE extract (75 μL), feeding solution (30 μL of 5x concentrated feeding solution) and ultra-pure water free of DNases and RNases (ThermoFisher Australia Cat# 10977015). Reactions were started by adding plasmid DNA, and were incubated for 2.5 h at 25 °C. Cell-free in vitro biocatalytic conversion of chrysoeriol to Cfl15 or Cfl20 [000190] Conversion of chrysoeriol to Cfl15 or Cfl20 by CsPT3_trunc was performed using the LTE system. Cell free protein synthesis reactions were prepared containing pCellFree / CsPT3_trunc plasmid DNA, or an equivalent volume of water as a negative control. Chrysoeriol (0.1 mM), FPP or GGPP (0.2 mM) and magnesium chloride (10 mM) were added 2.5 hours after initiation of cell-free protein synthesis, and reactions were incubated at 25 °C with sampling at 0- 30-, 60-, and 120-min. Reactions were quenched by addition of 3 volumes of ice-cold ethyl acetate and thorough vortex mixing. Quenched samples were shaken at 37 °C for 1 h at 400 rpm followed by centrifugation at 16000 g for 15 min. The supernatant was then transferred to a new tube and dried under a stream of nitrogen gas. Dried samples were stored at -20 °C until subsequent analysis. Reaction products corresponding to the predicted mass of Cfl15 were observed for the reaction with FPP (Figure 12).In vitro biocatalytic conversion of luteolin to Cfl15 or Cfl20 using E. coli lysates [000191] Coding sequences for CsPT3 and CsPT3_trunc were cloned into the pET29b expression vector individually (Table 3). Table 3: E. coli expression plasmids.[000192] Plasmids were transformed into E. coli BL21(DE3). Cell lysates containing CsPT3 or CsPT3_trunc enzymes were prepared as follows: starter cultures in 5 mL LB medium (10 g bacto-tryptone / L, 5 g yeast extract / L, 10 g sodium chloride / L, glucose (0.2 %, w / v), and 50 mg kanamycin / L) were inoculated from glycerol stocks and incubated for 16 h at 37 °C with shaking at 180 rpm. Starter cultures were diluted 1 in 50 into 50 mL terrific broth medium (20 g bacto- tryptone / L, 24 g yeast extract / L, 0.4 % (v / v) glycerol, 17 mM potassium phosphate monobasic, 72 mM potassium phosphate dibasic) in conical flasks and incubated at 37 °C with shaking at 180 rpm. Upon reaching OD600 nm = 0.6, gene expression was induced by addition of isopropyl β-D-1-thiogalactopyranoside (1 mM). Cultures were transferred to 18 °C and incubated with shaking (180 rpm) for a further 18 h. Cells were sedimented by centrifugation and washed twice in phosphate buffered saline, before resuspending in lysis buffer (50 mM Tris-Cl, pH 7, 10 mg DNase / mL, and AEBSF protease inhibitor cocktail (1 mM, Thermo Scientific). Resuspended cells were lysed using a Constant Systems Cell Disruptor at 30 kpsi. Lysates were clarified by centrifugation at 20,000 g. Prenyltransferase assays were performed by incubating 0.2 mL or 0.5 mL lysate (containing ~0.05 mg total protein / mL) with chrysoeriol (0.2 mM), FPP or GGPP (0.2 mM) and magnesium chloride (5 mM) at 37 °C for 1 h. Preliminary cannflavin-15 analytical methods [000193] Metabolites were identified via liquid chromatography-mass spectrometry. No analytical standard is available for Cfl15 or Cfl20, so identification of product formation was based on expected parent ion mass. Analyses were performed using liquid chromatography–mass spectrometry (LC–MS) on a Shimadzu Nexera X2 UPLC with 8050 triple quadrupole detector (Shimadzu, Japan) equipped with a C18 column (Phenomenex, Kinetex 2.6 µm C18, 100 x 2.1mm). The mobile phase comprised ultrapure water and acetonitrile with 0.1% formic acid. Metabolites were eluted with a reverse-phase gradient mobile phase 65% acetonitrile increasing to 95% acetonitrile over 10 min at a flow rate of 0.5 mL / min. After 10 min the column was re- equilibrated in 65% acetonitrile for 3 min. The MS instrument was operated positive ionizationmode with selected ion monitoring (SIM) for the relevant characteristic parent ion masses (Cfl15, m / z = 505). Analytical methods - metabolite identification [000194] Metabolites were identified via HPLC (Shimadzu HPLC with diode array detector) and LC-MS (Shimadzu Nexera X2 UPLC with 8050 triple quadrupole detector). All liquid chromatography methods used a Phenomenex 5μm C18, 150 x 4.6 mm stationary phase column, mobile phase A consisting of water with 0.1 % formic acid, and mobile phase B consisting of acetonitrile with 0.1% formic acid. Due to substantial differences in product retention time, different elution gradients were used for each cannflavin product as follows. Cannflavin A: 0-15 min linear gradient from 60-80% acetonitrile, 15.01-20 min isocratic elution in 60% acetonitrile. Cannflavin B: 0-15 min isocratic elution in 50% acetonitrile. Cannflavin 15 and cannflavin 20: 0-15 min linear gradient from 65-95% acetonitrile, 15.01-20 min isocratic elution in 65% acetonitrile. Chrysoeriol, cannflavin A, and cannflavin B were validated with comparison to authentic standards. In LC-MS analysis, the MS was operated positive ionization mode with selected ion monitoring (SIM) for the relevant characteristic parent ion masses (chrysoeriol, m / z = 301; cannflavin A, m / z = 437; cannflavin B, m / z = 369; Cfl15, m / z = 505; Cfl20, m / z = 573). Prediction of physicochemical properties [000195] Predicted octanol-water partition coefficients and molecular volumes were predicted using ChemDraw version 22.2 (PerkinElmer) based on fragments reported previously. Preparative scale biosynthesis of cannflavin-15 [000196] A 20 mL seed culture in LB medium (10 g tryptone / L, 5 g yeast extract / L, 10 g sodium chloride / L) plus kanamycin (50 μg / mL) was inoculated with E. coli BL21(DE3) bearing pET29b / CsPT3_trunc. The seed culture was incubated for 16 h at 37 °C with shaking at 180 rpm.10 mL of seed culture was inoculated into 1 L of terrific broth (12 g tryptone / L, 24 g yeast extract / L, 9.4 g dipotassium hydrogen phosphate, 2.2 g potassium dihydrogen phosphate, 0.4% glycerol) plus kanamycin (50 μg / mL) and incubated at 37 °C for 2.5 h with agitation at 180 rpm. Expression was induced by the addition of isopropyl β-D-1-thiogalactopyranoside (300 μM) and incubated for a further 24 h at 18 °C, 180 rpm. The culture was harvested via centrifugation and the cell pellet was resuspended in 50 mL lysis buffer (50 mM Tris-Cl, pH 9.0, 10 mM sodium chloride, 1 cOmpleteTMmini EDTA-free protease inhibitor tablet (Roche, Merck catalogue number 04693159001). Cells were lysed with a single pass at 25 kpsi in a Constant Systems cell disruptor. The crude lysate was centrifuged at 20,000 g for 20 min at 4 °C.[000197] Clarified lysate was diluted to a final volume of 87 mL with 50 mM Tris-Cl, pH 9.0, 10 mM sodium chloride, and addition of magnesium chloride (5 mM), chrysoeriol (500 μM), and trans,trans-FPP (150 μM). The reaction was incubated in a water bath at 37 °C for 2 h, and then quenched by addition of formic acid to a final concentration of 1%. [000198] Reaction products were extracted by vigorous mixing with an equal volume of ethyl acetate. The solvent phase was separated via centrifugation at 3000 g for 10 min. After removing the solvent phase, the ethyl acetate extraction was repeated and all ethyl acetate fractions were pooled and dried under a continuous stream of nitrogen. [000199] The dried sample was resuspended in a total of 3.6 mL acetonitrile and separated on a Phenomenex Kinetix 5 μM C18250 x 4.6 mm stationary phase column with the following mobile phase gradient: 0-15 min linear gradient from 65-95% acetonitrile with 0.1 % formic acid, 15.01-20 min isocratic elution in 65% acetonitrile with 0.1 % formic acid. A preliminary 10 µL injection indicated the presence of a cannflavin-15 peak with an elution time of approximately 11.5-12 min. This fraction was collected in multiple sample injections, pooled, and dried under a continuous stream of nitrogen. The recovered product mass was estimated at 1.1 mg by comparison of a reinjected sample with a cannflavin B standard curve (Figure 14). Characterisation of cannflavin-15 structure via nuclear magnetic resonance [000200] NMR experiments were recorded on a Bruker 600 Avance III HD spectrometer equipped with a BBO-Probe (5 mm) with z-gradient (1H: 600.13 MHz;13C: 150.90 MHz) using standard Bruker pulse sequences:1H (zg30),13C (DEPTQ135; deptqgpsp), COSY (cosygpmfqf), TOCSY (mlevphpp), HSQC (hsqcedetgpsisp2.3), HMBC (hmbcetgpl3nd) and NOESY (noesygpphpp). Cannflavin samples were prepared in acetone-d6 and spectra were recorded at 25 °C. Chemical shifts (δ) are reported as parts per million (ppm) relative to the residual solvent signals and coupling constants are stated in Hertz (Hz). Signal multiplicities are quoted as follows: s = singlet, bs = broad singlet, d = doublet, t = triplet, q = quartet, p = pentet and m = multiplet. NMR spectra were analysed in Bruker TopSpin 4.0.6. Results and discussion Biocatalytic synthesis of novel cannflavins from chrysoeriol [000201] In preliminary experiments, a product compound corresponding to the predicted Cfl15 molecular weight (505 g / mol) was identified in incubations of FPP and chrysoeriol with CsPT3_trunc, both when the enzyme was expressed in the Leishmania cell free protein expression system and in lysates of E. coli that expressed CsPT3_trunc (Figure 13). Productformation was not observed in negative controls where DNA encoding the CsPT3_trunc enzyme was omitted from either expression system. [000202] Further incubations of chrysoeriol with CsPT3_trunc and FPP or GGPP confirmed product formation with both alternative prenyl diphosphate donors, with molar masses corresponding to new cannflavin molecules with 15- or 20-carbon prenyl tails (Figure 4). [000203] Extension of the prenyl tail to 15 or 20 carbons significantly alters the volume of the molecule and net hydrophobicity described by the octanol-water partition coefficient (Table 4). This change in the logPoct / water can enable different modes of delivery and different tissue distribution as shown by the properties of comparison drugs having similar molecular masses and and logPoct / watervalues in Table 4. Table 4: Properties of natural and novel cannflavins compared with drugs having similar molecular masses and logPoct / water values.[000204] Reaction and substrate promiscuity is recognised among flavone prenyltransferase enzymes, and the degree of promiscuity varies widely between individual enzymes. The GuA6DT prenyltransferase from Glycyrrhiza uralensis catalyses 6-prenylation of several flavones including apigenin, luteolin, and chrysoeriol with DMAPP. GuA6DT also accepts GPP as a prenyl diphosphate donor but with catalytic rates less than 1% of that observed withDMAPP, and no prenylation reactions were observed when incubated with FPP or GGPP. A highly promiscuous prenyltransferase from Streptomyces sp. CL190 (NphB) catalyses prenylation of a wide range of flavone substrates at multiple positions with GPP but not with DMAPP. Substrates of NphB include naringenin which it can geranylate at two positions, resulting in 6-geranyl naringenin and 7-O-geranyl naringenin, where the 7-O-geranyl naringenin is the primary product. In incubations with chrysoeriol and GPP, NphB also produces 6-geranyl chrysoeriol (cannflavin A) as a minor product while the major product is presumably 7-O- geranyl chrysoeriol. Preparative scale synthesis of Cannflavin-15 [000205] Reaction conditions were optimised with the aim of producing Cfl15 for structural characterisation. Product formation Cfl15 product formation peaked after 120 min incubation with CsPT3_trunc and chrysoeriol (Figure 5). Reaction rates and total product formation were greater when reactions were incubated with mixed isomers of synthetic FPP than when pure trans,trans-FPP was used, suggesting that multiple isomers of FPP may be accepted by CsPT3_trunc. To synthesise a pure product for structural characterisation, trans,trans-FPP was used. [000206] A single preparative scale incubation was undertaken where CsPT3_trunc (from a 1 L E. coli culture lysed and concentrated) was combined with 5 mg trans,trans-FPP and 13 mg chrysoeriol in a total volume of 87 mL. A single reaction product was identified and approximately 1.1 mg material was isolated via preparative HPLC (Figure 6). Cannflavin-15 structural characterisation [000207] The structure of cannflavin 15 (Figure 7) was confirmed by NMR spectroscopy. Full structural elucidation was performed on the basis of1H and13C (DEPTQ135) spectroscopy, and 2D correlation spectroscopy (COSY, TOSCY, HSQC, HMBC, NOESY). Spectral assignments were supported by comparison with experimental and published spectroscopic data for cannflavin A (Table 5). [000208]1H NMR (600.13 MHz, acetone-d6) δ = 13.30 (s, 1H), 7.61 (d, 2.2 Hz, 1H), 7.58 (dd, 8.3, 2.2 Hz, 1H), 7.00 (d, 8.3 Hz, 1H), 6.69 (s, 1H), 6.62 (s, 1H), 5.30 (tq, 7.3,1.2 Hz, 1H), 5.09 (tq, 7.1,1.3 Hz,, 1H), 5.04 (tp, 7.1, 1.4 Hz, 1H), 3.99 (s, 3H), 3.38 (d, 7.3 Hz, 2H), 2.08 (m, 2H), 1.98 (m, 2H), 1.96 (m, 2H), 1.89 (m, 2H), 1.81 (d, 1.1 Hz, 3H), 1.59 (d, 1.1 Hz, 3H), 1.56 (s, 3H), 1.54 (s, 3H).13C NMR (150.90 MHz, acetone-d6) δ = 183.10, 164.63, 162.41, 160.23, 156.60, 151.25, 148.82, 135.45, 135.18, 131.5, 125.19, 124.93, 123.77, 123.26, 121.24, 116.30,112.29, 110.48, 105.2, 104.47, 94.18, 56.59, 40.49, 40.45, 27.41, 27.13, 25.83, 21.97, 17.68, 16.25, 16.11. Table 5: Chemical structure of cannflavin-15 and table of NMR assignmentsa: Identified from HMBC data;*:Cannflavin A assignments from Rea, K. A. et al., Biosynthesis of cannflavins A and B from Cannabis sativa L., Phytochemistry, 164, 162–171 (2019).[000209] The1H spectrum of cannflavin-15 (Figures 8–9) was analogous to that of cannflavin A, with extra resonances observed in the vinylic and aliphatic regions due to the farnesyl side chain (rather than the geranyl chain of cannflavin A). The five resonances in the aromatic region (Figure 8) are consistent with the A, B, C ring substitution pattern in cannflavin 15. Signals at 7.61, 7.58 and 7.00 ppm exhibited J-coupling indicative of the 1,3,5-substitution pattern in ring B, supported by COSY, and were assigned to H-2’, H-6’ and H-5’ respectively. The substitution pattern in ring B was further confirmed by HMBC and NOESY spectroscopy, with O-CH3 (3.99 ppm) correlating with H-2’ in the latter (Figures 5, 6). [000210] Direct observation of the13C resonances in the multiplicity edited DEPTQ135 spectrum (Figure 10) was achieved for all C sites, despite low signal to noise, except for C-4a and C-11” which were each clearly identified by several HMBC correlations. Assignment of the13C resonances was aided by the multiplicity edited HSQC spectrum. [000211] The singlets at 6.69 and 6.62 ppm were assigned to H-3 and H-8 respectively, on the basis of weak long-range correlation of H-8 with H-1” in the TOCSY spectrum, clear NOESY interactions between H-3 and H-2’ and 6’ of ring B, and a range of HMBC correlations (Figures 11, 12). [000212] In the low field region of the1H spectrum (Figure 3), resonances due to the methyl groups of the farnesyl side chain are evident at 1.81 (H-15”), 1.59 (H-12”), 1.56 (H-14”) and 1.54 (H-13”) ppm. While these approximate singlets, under closer observation they exhibit weak J-coupling consistent with long range coupling in the tri-substituted double bonds of the prenyl residues. The vinylic protons of the farnesyl chain are observed at 5.30 (H-2”), 5.09 (H-6”) and 5.04 (H-10”) ppm. The methylene resonance of H-1” is well-resolved at 3.38 ppm, while those in the region 2.10 – 1.86 ppm (H-5”, H-4”, H-9”, H-8”) are partially overlapped, with H-5” obscured by the residual solvent peak. Full assignment of the farnesyl side-chain was facilitated by COSY and TOCSY, with the TOCSY spectrum clearly differentiating each prenyl residue. NOESY spectroscopy confirmed the connectivity in the chain and stereochemistry of the vinylic groups (Figure 12). [000213] The 5-OH is observed as a sharp peak at 13.30 ppm in the1H spectrum, consistent with the phenol undergoing intramolecular H-bonding with the neighbouring carbonyl oxygen. Distinct peaks are not observed for 4’-OH and 7-OH, presumably due to broadening associated with H-bonding with adventitious water in the NMR solvent. A very broad resonance at 8.2-10.0 ppm was tentatively assigned to these phenols, but it was not possible to confirm this assignment through correlation experiments. NOESY indicated chemical exchange between the broad signal and water, which is consistent with the phenol assignment but not conclusive. Similarly, 4’-OHand 7-OH are not reported in the published spectra of cannflavins A and B1,2,3 but we observe an identical broad signal in our experimental data for cannflavin A. [000214] The substitution pattern around rings A and C was confirmed by a host of HMBC correlations (Figure 5). Positioning of the farnesyl chain at C-6 was supported by correlations between 5-OH and C-6, as well as H-1” and C-6. Similarly, correlations between H-1” and both C-5 and C-7, but not C-8 or C-8a, confirms substitution at C-6 rather than C-8. [000215] The NOESY spectrum indicates a short-range interaction between H-8 and water in solution (2.81 ppm), presumably due to long-lived (on the NMR timescale) hydrogen bonding interactions between the water and 7-OH and / or O-1 in ring C. In addition to short-rangeinteractions, NOESY spectroscopy also provides information on chemical and conformational exchange. In the NOESY spectrum of cannflavin 15, 5-OH undergoes proton exchange with water. Similarly, the broad resonance at 8.2-10.0 ppm, tentatively assigned to 4’-OH and 7-OH, shows proton exchange with water. Virtual Prostaglandin E Synthase 2 docking [000216] Three dimensional structures of the the ligands: cannflavin A, cannflavin B, cannflavin 15, and cannflavin 20, were prepared by manually building the two dimensional structures in ChemDoodle (v12.7.0, iChemLabs) and exporting them as SDF files. The resulting three dimensional structures were manually inspected in Biovia Discovery Studio Visualizer (v21.1.0.20298) and errors that occurred during the conversion to SDF format were corrected where necessary (for example, misplacement of double bonds that occurred in the conversion process). [000217] Docking was performed using GPU-enabled GNINA (v1.1.0) installed as a module on a GPU cluster. Whole-protein mode was used to cover the entirety of each structure with no bounding-box restrictions. All ligands were, singly, docked into Prostaglandin E Synthase 2. The docked structures of all four (superimposed) ligands are shown in Figure 16. The ligands are coloured blue, and the reactive heme site of Prostaglandin E Synthase 2 is shown in tan. The docking and affinity scores for each of the ligands are set out in table 6 below. Table 6: CNN docking scores for prostaglandin synthase E2.[000218] Although the invention has been described with reference to specific examples, it will be appreciated by those skilled in the art that the invention may be embodied in many other forms. In particular, features of any one of the various described examples may be provided in any combination in any of the other described examples. Various modifications and alterations to this invention will become apparent to those skilled in the art without departing from the scope and spirit of this invention. It should be understood that this invention is not intended to be unduly limited by the illustrative embodiments and examples set forth herein and that such examples and embodiments are presented by way of example only.

Claims

Claims:

1. A compound having the structure of Formula (I):, or a pharmaceutically acceptable salt and / or hydrate or solvate thereof; wherein: R1is H or OCH3; R2is tri-prenyl, tetra-prenyl, penta-prenyl, hexa-prenyl, hepta-prenyl, or octa-prenyl; and R3is H.

2. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of claim 1, wherein R1is OCH3.

3. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of claim 1 or 2, wherein: R2is tri-prenyl, tetra-prenyl, penta-prenyl, or hexa-prenyl.

4. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 3, wherein: R2is tri-prenyl, or tetra-prenyl.

5. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 4, which has a structure selected from the following:,, and pharmaceutically acceptable salts and / or hydrates or solvates thereof.

6. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 5, which has the following structure:, or a pharmaceutically acceptable salt and / or hydrate or solvate thereof.

7. A pharmaceutical composition comprising the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 6 and a pharmaceutically acceptable excipient.

8. The pharmaceutical composition of claim 7, wherein the composition is suitable for topical administration.

9. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 6, or the pharmaceutical composition of claim 7 or claim 8 for use in treating inflammation in a subject in need thereof, optionally wherein the inflammation is treatable by modulation of prostaglandin E2 synthetase 1 (mPGES-1) and / or arachidonic acid 5-lipoxygenase (5-LO).

10. Use of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 6, or the pharmaceutical composition of claim 7 or claim 8, for the manufacture of a medicament for treating inflammation in a subject in need thereof, optionally wherein the inflammation is treatable by modulation of prostaglandin E2 synthetase 1 (mPGES- 1) and / or arachidonic acid 5-lipoxygenase (5-LO).

11. A method of treating inflammation in a subject in need thereof, said method comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptablesalt and / or hydrate or solvate thereof of any one of claims 1 to 6, or the pharmaceutical composition of claim 7 or claim 8, to the subject, optionally wherein the inflammation is treatable by modulation of prostaglandin E2 synthetase 1 (mPGES-1) and / or arachidonic acid 5-lipoxygenase (5-LO).

12. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 6, or the pharmaceutical composition of claim 7 or claim 8 for use in treating cancer in a subject in need thereof, optionally wherein the cancer is treatable by modulation of tropomyosin receptor kinase B (TrkB).

13. Use of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 6, or the pharmaceutical composition of claim 7 or claim 8, for the manufacture of a medicament for treating cancer in a subject in need thereof, optionally wherein the cancer is treatable by modulation of tropomyosin receptor kinase B (TrkB).

14. A method of treating cancer in a subject in need thereof, said method comprising administering a therapeutically effective amount of the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 6, or the pharmaceutical composition of claim 7 or claim 8, to the subject, optionally wherein the cancer is treatable by modulation of tropomyosin receptor kinase B (TrkB).

15. The compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of claim 12, use of claim 13, or method of claim 14, wherein the cancer is selected from the group consisting of transitional cell carcinoma, bladder, brain, breast, Kaposi sarcoma, leukemia, lung, melanoma, ovarian, pancreatic, colon, and prostate cancer.

16. A method of producing the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof of any one of claims 1 to 6, said method comprising the following steps: preparing a reaction mixture comprising: chrysoeriol or apigenin, a poly-prenyl pyrophosphate, a prenyltransferase or a nucleic acid encoding a prenyltransferase; and incubating the reaction mixture under conditions suitable for the prenyltransferase to catalyse prenylation of the chrysoeriol or apigenin using the poly-prenyl pyrophosphate as a prenyl donor to thereby produce the compound or pharmaceutically acceptable salt and / or hydrate or solvate thereof.

17. The method of claim 16, wherein the prenyltransferase catalyses prenylation of chrysoeriol or apigenin at the 6-position thereof.

18. The method of claim 16 or claim 17, wherein the reaction mixture further comprises a cell lysate.

19. The method of claim 18, wherein the cell lysate is derived from cells engineered to express the prenyltransferase.

20. The method of claim 18 or claim 19, wherein the reaction mixture comprises a nucleic acid encoding the prenyltransferase.

21. The method of claim 20, wherein the cell lysate is a cell-free protein expression lysate.

22. The method of claim 20 or claim 21, wherein the reaction mixture comprises RNA encoding the prenyltransferase, and wherein the reaction mixture is incubated under conditions suitable for the RNA to be translated to produce the prenyltransferase.

23. The method of claim 20 or claim 21, wherein the reaction mixture comprises DNA encoding the prenyltransferase, and wherein the reaction mixture is incubated under conditions suitable for the DNA to be transcribed to RNA and for the RNA to be translated to produce the prenyltransferase.

24. The method of any one of claims 18 to 23, wherein the cell lysate is an E. coli cell lysate or a Leishmania cell lysate.

25. The method of any one of claims 16 to 24, wherein the prenyltransferase comprises the sequence set forth in SEQ ID NO: 1, or a sequence having at least about 80% identity to SEQ ID NO:

1.

26. The method of any one of claims 16 to 25, wherein the prenyltransferase lacks a chloroplast transit peptide sequence.

27. The method of claim 26 wherein the chloroplast transit peptide sequence comprises the sequence set forth in SEQ ID NO: 3, or a sequence having at least about 80% identity to SEQ ID NO:

3.

28. The method of claim 26 or claim 27, wherein the prenyltransferase is between about 300 and 350 amino acids in length.

29. The method of any one of claims 26 to 28, wherein the prenyltransferase comprises the sequence set forth in SEQ ID NO: 2, or a sequence having at least about 80% identity to SEQ ID NO: 2.

30. The method of any one of claims 16 to 29, wherein the prenyltransferase comprises one or more mutations at a leader peptide cleavage site.

31. The method of any one of claims 16 to 30, wherein the prenyltransferase is Cannabis sativa PT3 or a variant thereof.

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