Manufacturing process for glucosinolate

Abstract

The present invention relates to a manufacturing process for glucosinolate, or a derivative, analog, stereoisomer or salt thereof, comprising flowing a stream comprising a thiolated compound from a first reactor into a solution comprising a halo oxime in a second reactor in order to form a thiohydroximate compound, and reacting the thiohydroximate compound with pyridine sulfur trioixde in the second reactor in order to form a sulfated thiohydroximate compound.

Description

[0001] Manufacturing Process for Glucosinolate

[0002] Technical Field

[0003] The present invention relates, in general terms, to a manufacturing process for glucosinolate.

[0004] Background

[0005] Glucosinolates are a group of naturally occurring compounds found in plants, particularly in the Brassicaceae family, which includes vegetables such as broccoli, cabbage, cauliflower, and kale. These compounds are responsible for the distinctive flavours and health benefits associated with these vegetables.

[0006] Glucosinolates are composed of a glucose molecule attached to a sulfur-containing side chain and a variable R-group. When the plant tissue is damaged, such as during chewing or cutting, the enzyme myrosinase is released and catalyses the breakdown of glucosinolates into various bioactive compounds, including isothiocyanates, nitriles, and thiocyanates. Isothiocyanates, in particular, have been shown to possess anti-cancer, anti-inflammatory, and antioxidant properties, and may play a role in reducing the risk of certain chronic diseases, such as cancer and cardiovascular disease.

[0007] Glucosinolates and their breakdown products have also been studied for their potential in agriculture, as they can act as natural pesticides and herbicides, contributing to the plant's defence against pests and pathogens.

[0008] However, glucosinolates are currently directly extracted from plants, or produced via biosynthesis. Such processes are complex and costly.

[0009] While some laboratory scale synthesis of glucosinolate exist, transitioning a laboratory process to industrial-scale production presents a complex array of technical, economic, and operational challenges. A fundamental prerequisite for successful scale-up is a comprehensive understanding of the synthesis process to facilitate optimisation, selection of appropriate industrial equipment capable of replicating laboratory conditions and scheduling of tasks to be performed on each corresponding equipment in an optimal manner. These technical and operational hurdles are non-trivial.

[0010] It would be desirable to overcome or ameliorate at least one of the above-describedproblems.

[0011] Summary

[0012] The present disclosure concerns a method of manufacturing glucosinolate, or a derivative, analog, stereoisomer or salt thereof, comprising:

[0013] a) flowing a stream comprising a compound of Formula (I) from a first reactor into a solution comprising a compound of Formula (II) in a second reactor in order to form a compound of Formula (III); and

[0014]

[0015] b) reacting the compound of Formula (III) with pyridine sulfur trioxide in the second reactor in order to form a compound of Formula (IV);

[0016]

[0017] wherein

[0018] Ri is selected from H;

[0019] each R2 is independently selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted acyl, and optionally substituted silyl;

[0020] Rs is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; and

[0021] Xi is halo.

[0022] In some embodiments, a mole ratio of compound of Formula (I) to compound of Formula (II) is about 1:1.02 to about 1:1.1.

[0023] In some embodiments, step a) further comprises adding ethyl acetate to the second reactor before flowing the stream comprising a compound of Formula (I) into the second reactor.In some embodiments, step a) further comprises adding a carbonate to the second reactor in order to precipitate the compound of Formula (III).

[0024] In some embodiments, the compound of Formula (III) is concentrated to less than about 0.5 times its initial volume.

[0025] In some embodiments, the compound of Formula (III) is filtered, washed, and / or dried.

[0026] In some embodiments, step a) further comprises reacting a compound of Formula (V) with sodium sulfite (sulphite) in the first reactor in order to form the compound of Formula (I):

[0027]

[0028] wherein F is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl.

[0029] In some embodiments, a mole ratio of compound of Formula (V) and sodium sulfite is about 1:1 to about 1:1.5.

[0030] In some embodiments, the reaction is performed for a duration of at least 1 h.

[0031] In some embodiments, reaction is performed in a biphasic mixture of an aqueous medium and ethyl acetate

[0032] In some embodiments, the ethyl acetate is flowed into the second reactor.

[0033] In some embodiments, the method further comprises a step before step a) of synthesising the compound of Formula (V) in the first reactor.

[0034] In some embodiments, the compound of Formula (V) is synthesised by reacting a compound of Formula (VI) with thiourea of Formula (VII):

[0035]

[0036] wherein X2 is halo.

[0037] In some embodiments, a mole ratio of compound of Formula (VI) to thiourea of Formula (VII) is about 1:1 to about 1:2.

[0038] In some embodiments, the reaction is performed at a temperature of about 60 °C to about 90 °C for a duration of about 0.5 h to about 5 h in order to precipitate compound of Formula (V).

[0039] In some embodiments, the compound of Formula (V) is filtered, washed and / or dried.

[0040] In some embodiments, the first reactor is characterized by a volume of at least 100L.

[0041] In some embodiments, step a) further comprises reacting a compound of Formula (VIII) with N-chlorosuccinimide in order to form compound of Formula (II);

[0042]

[0043] In some embodiments, the reaction is performed at ambient temperature for a duration of about 1 h to about 5 h.

[0044] In some embodiments, the reaction is performed in ethyl acetate.

[0045] In some embodiments, the method further comprises a step before step a) of synthesising the compound of Formula (VIII) in the second reactor.

[0046] In some embodiments, the reaction is performed at ambient temperature for a duration of about 1 h to about 5 h.

[0047] In some embodiments, the reaction is distilled in order to precipitate the compound of Formula (VIII).In some embodiments, the compound of Formula (VIII) is filtered, washed and / or dried.

[0048] In some embodiments, the step of step of synthesising the compound of Formula (V) in the first reactor is concurrent with the step of synthesising the compound of Formula (VIII) in the second reactor.

[0049] In some embodiments, the second reactor is characterized by a volume of at least two times the volume of the first reactor.

[0050] In some embodiments, a mole ratio of compound of Formula (III) to pyridine sulfur trioxide is about 1:1.4 to about 1:2.

[0051] In some embodiments, step b) further comprises adding pyridine.

[0052] In some embodiments, a mole ratio of compound of Formula (III) to pyridine is about 1:0.3 to about 1:0.7.

[0053] In some embodiments, the reaction is performed at a temperature of about 60 °C to about 90 °C for a duration of about 0.5 h to about 5 h.

[0054] In some embodiments, step b) further comprises exchanging a cation associated with compound of Formula (IV).

[0055] In some embodiments, the compound of Formula (IV) is purified by adding an aqueous medium.

[0056] In some embodiments, step b) further comprises precipitating the compound of Formula (IV).

[0057] In some embodiments, the compound of Formula (IV) is filtered, washed, dried and / or recrystallized.

[0058] In some embodiments, the method further comprises a step after step b) of hydrolysing compound of Formula (IV) such that R2 is H.

[0059] In some embodiments, the method is configured to produce at least 5 kg of compound of Formula (IV).In some embodiments, the method is characterised by a duration of about 30 h.

[0060] The present disclosure also concerns a system for manufacturing glucosinolate, or a derivative, analog, stereoisomer or salt thereof comprising:

[0061] a) a first reactor;

[0062] b) a second reactor in fluid communication with the first reactor via a first one way valve;

[0063] c) filtration means in fluid communication with the first reactor via a second one way valve and separately in fluid communication with the second reactor via a third one way valve;

[0064] d) drying means in fluid communication with the filtration means and separately in fluid communication with the first and second reactors; and

[0065] e) a spray dryer in fluid communication with the first reactor.

[0066] Brief description of the drawings

[0067] Embodiments of the present invention will now be described, by way of non-limiting example, with reference to the drawings in which:

[0068] Figure 1 shows an exemplary process for synthesising glucosinolate.

[0069] Figure 2 shows step 1 of a block flow diagram for synthesising an oxime intermediate.

[0070] Figure 3 shows step 2 of a block flow diagram for synthesising a glucose isothiourea intermediate.

[0071] Figure 4 shows step 3 of a block flow diagram for synthesising a thiohydroximate intermediate.

[0072] Figure 5 shows step 4 of a block flow diagram for synthesising a sulphated thiohydroximate intermediate.

[0073] Figure 6 shows step 5 of a block flow diagram for synthesising glucosinolate.

[0074] Figure 7 shows a Gantt Chart for synthesising glucosinolate.

[0075] Figure 8 shows a process flow diagram for synthesising glucosinolate.

[0076] Detailed description

[0077] To overcome at least one of the technical and / or operational hurdles, a chemical and process engineering approach is employed, involving the development of block flow diagram (BFD) and process flow diagram (PFD) for the laboratory-scale GSL production process, calculation of process mass balance and matching tasks with equipment basedon the production recipe. A thorough evaluation of both capital and operational costs is conducted to ensure the economic viability of the scale-up endeavour. Several process improvements have also been incorporated into the synthesis.

[0078] In certain embodiments, the scaling up a laboratory-scale production of glucosinolate (GSL) using the oxime synthesis pathway from ten grams to an industrial kilogram scale is demonstrated, with a target production of 2000 kg per year. This was achieved through an assessment of both technical and economic aspects of the process.

[0079] Advantageously, the glucosinolate may be prepared as a spray-dried powder, or as an aqueous solution.

[0080] The present disclosure concerns a method of manufacturing glucosinolate, or a derivative, analog, stereoisomer or salt thereof, comprising:

[0081] a) flowing a stream comprising a compound of Formula (I) from a first reactor into a solution comprising a compound of Formula (II) in a second reactor in order to form a compound of Formula (III); and

[0082]

[0083] (I) (II) (HI) b) reacting the compound of Formula (III) with pyridine sulfur trioxide in the second reactor in order to form a compound of Formula (IV);

[0084]

[0085] wherein

[0086] Ri is selected from H;

[0087] each R2 is independently selected from H, optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted acyl;

[0088] R3 is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; and

[0089] Xi is halo.As used herein, a "derivative" is a compound that is derived from a similar compound by a chemical reaction. In this regard, the method may further comprise additional step(s) in order to derivatise the compound.

[0090] As used herein, an "analog" is a compound which shares the same parent molecule but differ by the addition or substitution of other atoms.

[0091] The stream from the first reactor comprising compound of Formula (I) may flow into the second reactor via a one-way valve. The second reactor is also appropriately sized such that it has sufficient volume to contain the stream from the first reactor. It was found that the manufacturing method may be completed using a combination of two reactors.

[0092] In some embodiments, each R2 is independently selected from H, and optionally substituted acyl. In some embodiments, each R2 is independently selected from a protecting moiety.

[0093] A protective group is a reversibly formed derivative of an existing functional group in a molecule. The protective group is temporarily attached to decrease reactivity so that the protected functional group does not react under synthetic conditions to which the molecule is subjected in one or more subsequent steps. As an example, whereas amines are nucleophiles and react with electrophiles, the amino group is no longer nucleophilic after being converted to a carbamate. Protecting an amine as a carbamate therefore enables other functional groups to undergo selective reactions with electrophiles whereby the carbamate (protected amino group) is left intact. However, two additional synthetic steps are needed to achieve this protection: the step to form the protected intermediate and a deprotection once the additional selective synthetic steps have been completed. In addition, the nature of the protective group must be chosen carefully to ensure adequate stability throughout all the intermediary synthesis steps. Moreover, the conditions for the protection and deprotection steps and the nature of the protective group itself mustn't interfere with other functional groups present in the molecule. If more than one functional group of the same type is present in a molecule, subtle differences in reactivity - for example caused by steric effects - can help to achieve the selective protection of just one functional group while another such functional group remains unprotected. Alternatively, a second such functional group could be protected with a different protecting group that has a different reactivity profile. Another opportunity is to build a larger molecule from subunits in which similar or identical functional groups have been differently protected beforehand. For example, a Boc-protected amino group can be deprotected in acidic media, whereas a Fmoc-protected amino group can be deprotected under basic conditions. The presence of both protective groups in the same molecule therefore enables selective deprotection of one protected amino group for a further reaction while the second protected amino group remains untouched.

[0094] In some embodiments, R2 is a hydroxy protecting group. By way of non-limiting example, hydroxyl protecting groups include methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxy methyl, bis(2-chloroethoxy) methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetra hydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, l-[(2-ch loro-4- methyl) pheny I] -4-methoxypi peridin-4-y I (CTMP), l,4-dioxan-2-yl, tetra hydrofuranyl, tetra hydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, l-(2-chloroethoxy)ethyl, 1-methyl-l-methoxyethyl, 1-methyl-l-benzyloxyethyl, l-methyl-l-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl, p,p'-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, o-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophtha I imidophenyl) methyl, 4, 4', 4"-tris( levu li noy loxy phenyl) methyl, 4,4' ,4"-tris( benzoyloxy pheny I) methyl, 3-( imidazol-l-y I) bis(4',4"-di methoxy pheny I) methyl, l,l-bis(4-methoxyphenyl)-l'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, l,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-l-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl) benzoate, 2-formyl benzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-( methylthiomethoxy) butyrate, 2-( methylthio methoxy methyl) benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2, 6-d ichloro-4-( 1, 1,3, 3-tetramethylbuty I) phenoxyacetate, 2,4-bis(l,l-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, (E)-2-methyl-2-butenoate, o-(methoxycarbonyl)benzoate, o-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

[0095] For example, hydroxyl may be protected using ether, silyl ether, ester moieties. For example, R2 may be tetrahydropyranyl, trimethylsilyl, methoxymethyl, acetyl (Ac), benzoyl (Bz), or pivaloyl (Piv). In some embodiments, each R2 is independently selected from acetyl (Ac), benzoyl (Bz), and pivaloyl (Piv).

[0096] In some embodiments, R3 is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl. In some embodiments, R3 is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl. In some embodiments, R3 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted methylsulfonylalkyl, optionally substituted methylsulfinylalkyl, and optionally substituted methylsulfenylalkyl, In some embodiments, R3 is phenyl.

[0097] In some embodiments, Xi is selected from F, Cl, Br, and I. In some embodiments, Xi is Br.Step a) is a step of forming a compound of Formula (III), which is a thiohydroximate compound.

[0098] In some embodiments, a mole ratio of compound of Formula (I) to compound of Formula (II) is about 1:1.02 to about 1:1.1. In other embodiments, the mole ratio is about 1:1.02 to about 1:1.09, about 1:1.02 to about 1:1.08, about 1:1.02 to about 1:1.07, about 1:1.02 to about 1:1.06, or about 1:1.02 to about 1:1.05.

[0099] In some embodiments, step a) further comprises adding a solvent to the second reactor before flowing the stream comprising a compound of Formula (I) into the second reactor. In some embodiments, the solvent is a polar aprotic solvent. The solvent may have a low to medium polarity. For example, the solvent may be pyridine, diethylamine, aniline, ethylacetate, dioxane, acetone, dichloroethane, tetrahydrofuran, dicholoromethane, chloroform, diethyl ether or a combination thereof. In some embodiments, the solvent is ethyl acetate. This avoids the reaction material becoming a solid mass, while maintaining the compound of Formula (III) as a slurry. It was found that high polar solvents will dissolve the compound of Formula (III), which is not desirable as side products are formed.

[0100] In some embodiments, a weight ratio of the additional solvent to the compound of Formula (I) is about 2: 1 to about 20:1. In other embodiments, the weight ratio is about 2:1 to about 18:1, about 2:1 to about 16:1, about 2:1 to about 14:1, about 2:1 to about 12:1, about 2:1 to about 10:1, about 2:1 to about 8:1, about 2:1 to about 6:1, or about 2: 1 to about 5:1.

[0101] In some embodiments, a volume ratio of the additional solvent to the solution in the second reactor is about 1:1 to about 1:5. In some embodiments, a volume ratio of the additional solvent to the solution in the second reactor is about 1:2.

[0102] In some embodiments, step a) further comprises adding a carbonate to the second reactor in order to precipitate the compound of Formula (III). The carbonate may be a metal carbonate. For example, potassium carbonate may be used.

[0103] The carbonate may be added in excess of the compound of Formula (III). In some embodiments, a mole ratio of compound of Formula (II) to carbonate is about 1:1.05 to about 1:1.5. In some embodiments, the mole ratio is about 1:1.2.In some embodiments, the carbonate is reacted with compound of Formula (III) at ambient temperature. In some embodiments, the temperature is about 10 °C to about 40 °C. In some embodiments, the carbonate is reacted with compound of Formula (III) for a duration of about 0.5 h to about 5h. In some embodiments, the carbonate is reacted with compound of Formula (III) for a duration of about 1 h.

[0104] In some embodiments, the compound of Formula (III) is concentrated. This may be done by concentrating a solution comprising compound of Formula (III) in a drying means, such as a rotary evaporator. The concentration may be performed until the solution is reduced to less than about 0.5 times its initial volume, or less than about 0.3 times its initial volume.

[0105] In some embodiments, the compound of Formula (III) is filtered. This may be performed in a filtration means, such as a filter press.

[0106] In some embodiments, the compound of Formula (III) is washed. The washing may be performed in a non-solvent. The non-solvent may be a polar solvent. For example, an aqueous medium and / or ethyl acetate may be used.

[0107] The term 'aqueous medium' used herein refers to a water based solvent or solvent system, and which comprises of mainly water. Such solvents can be either polar or nonpolar, and / or either protic or aprotic. Solvent systems refer to combinations of solvents which resulting in a final single phase. Both 'solvents' and 'solvent systems' can include, and is not limited to, pentane, cyclopentane, hexane, cyclohexane, benzene, toluene, dioxane, chloroform, diethylether, dichloromethane, tetra hydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, nitromethane, propylene carbonate, formic acid, butanol, isopropanol, propanol, ethanol, methanol, acetic acid, ethylene glycol, diethylene glycol or water. Water based solvent or solvent systems can also include dissolved ions, salts and molecules such as amino acids, proteins, sugars and phospholipids. Such salts may be, but not limited to, sodium chloride, potassium chloride, ammonium acetate, magnesium acetate, magnesium chloride, magnesium sulfate, potassium acetate, potassium chloride, sodium acetate, sodium citrate, zinc chloride, HEPES sodium, calcium chloride, ferric nitrate, sodium bicarbonate, potassium phosphate and sodium phosphate.

[0108] In some embodiments, the compound of Formula (III) is dried. This may be performed in a drying means, such as a drying oven. The drying may be performed at a temperature of about 50 °C to about 200 °C.Glucose Isothiourea Synthesis

[0109] The compound of Formula (I) may be synthesised in the first reactor. For example, a glucose isothiourea compound may be synthesized in the first reactor, following which the isothiourea moiety may be reduced to thiol using sodium sulfite (sulphite) and reacted with compound of Formula (II).

[0110]

[0111] In some embodiments, step a) further comprises reacting a compound of Formula (V) er to form the compound of Formula (I):

[0112]

[0113] wherein F is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl.

[0114] In some embodiments, F is selected from H, optionally substituted alkyl. In some embodiments, F is H.

[0115] The compound of Formula (V) may be in its salt form, such as hydrobromide or hydrochloride salt.

[0116] In some embodiments, the reaction is performed by adding the compound of Formula (V) into the first reactor containing sodium sulfite. The sodium sulfite may be dissolved in a biphasic mixture, comprising an aqueous medium and an organic solvent with a low dielectric constant. For example, ethyl acetate may be used.

[0117] In some embodiments, a mole ratio of compound of Formula (V) and sodium sulfite is about 1 : 1 to about 1:1.5. In some embodiments, a mole ratio of compound of Formula (V) and sodium sulfite is about 1:1. More sodium sulfite may be added if the glucose isothiourea is wet and acid still remains.In some embodiments, the reaction is performed in the absence of oxygen or under oxygen free conditions. In some embodiments, the biphasic mixture of sodium sulfite is purged with an inert gas (nitrogen and / or argon) before reacting with compound of Formula (V).

[0118] In some embodiments, the reaction is performed for a duration of at least 1 h.

[0119] In some embodiments, the organic solvent of the biphasic mixture is flowed into the second reactor. This may be done by allowing the biphasic mixture to phase separate and extracting only the organic phase. The compound of Formula (I) is contained within the organic phase.

[0120] The compound of Formula (V) (glucose isothiourea compound) may be synthesized from basic starting materials such as glucose.

[0121]

[0122] In some embodiments, the method further comprises a step before step a) of synthesising the compound of Formula (V) in the first reactor. The compound of Formula (V) may be synthesized using glucose, or a derivative, analog, stereoisomer, solvate or salt thereof and thiourea.

[0123] In some embodiments, the compound of Formula (V) is synthesised by reacting a compound of Formula (VI) with thiourea of Formula (VII):

[0124]

[0125] wherein X2 is halo.

[0126] In some embodiments, the compound of Formula (V) is synthesised by reacting a compound of Formula (Vl-i) with thiourea of Formula (VII):

[0127]

[0128] wherein X2 is halo.

[0129] When R2 is acetyl, the a anomer of tetraacetyl-D-glucopyranosyl halide of Formula (VI) gives the p-product as shown in Formula (V). Tetraacetyl-D-glucopyranosyl halide undergoes a spontaneous SNl-like loss of X', followed by internal reaction with the ester group at C2 to form an oxonium ion. Since the acetate at C2 is on the bottom of the glucose ring, the C-0 bond also forms from the bottom. Backside SN2 displacement of the oxonium ion then occurs with the usual inversion of configuration, yielding a p- product and regenerating the acetate at C2.

[0130]

[0131] In some embodiments, X2 is selected from Cl and Br. In some embodiments, X2 is Br.

[0132] In some embodiments, a mole ratio of compound of Formula (VI) to thiourea of Formula (VII) is about 1:1 to about 1:2. In some embodiments, a mole ratio of compound of Formula (VI) to thiourea of Formula (VII) is about 1:1.

[0133] In some embodiments, the reaction is performed at a temperature of about 60 °C to about 90 °C. In some embodiments, the reaction is performed at a temperature of about 80 °C.In some embodiments, the reaction is performed for a duration of about 0.5 h to about 5 h in order to precipitate compound of Formula (V). In some embodiments, the reaction is performed for a duration of about 1 h.

[0134] In some embodiments, the compound of Formula (V) is filtered. This may be performed in a filtration means, such as a filter press.

[0135] In some embodiments, the compound of Formula (V) is washed. The washing may be performed in a non-solvent. The non-solvent may be a polar solvent. For example, an aqueous medium and / or ethyl acetate may be used.

[0136] In some embodiments, the compound of Formula (V) is dried. This may be performed in a drying means, such as a drying oven. The drying may be performed at a temperature of about 50 °C to about 200 °C.

[0137] In some embodiments, when R2 is not H, compound of Formula (VI) is synthesized in the first reactor. The compound of Formula (VI) may be synthesized starting from glucose or a derivative, analog, stereoisomer, solvate or salt thereof.

[0138] In some embodiments, glucose is reacted with a protecting group precursor and hydrogen halide in order to form compound of Formula (VI). In some embodiments, the protecting group precursor is selected from acetic anhydride, dihydropyran, methoxymethyl chloride and trimethylsilyl chloride. The hydrogen halide may be HCI or HBr,. The protecting group precursor and / or hydrogen halide may be in excess of glucose. For example, the protecting group precursor and / or hydrogen halide may be in excess of glucose by about 0.5 times, 1 times, 1.5 times, 2 times or 4 times.

[0139] In some embodiments, the reaction is performed at a temperature of about 10 °C to about 60 °C. Cooling means may be required as the reaction is exothermic.

[0140] In some embodiments, the reaction is performed for a duration of about 1 h to about 8 h. In some embodiments, the reaction is performed for a duration of about 3 h.

[0141] If an excess of hydrogen halide is used, a neutralising agent may be added to neutralise excess hydrogen halide. For example, sodium acetate may be added. Sodium bromide may precipitate and be removed.

[0142] In some embodiments, the first reactor is characterized by a volume of at least IDOL.Oxime synthesis

[0143] The compound of Formula (II) may be synthesized in the second reactor. For example, compound of Formula (II) may be synthesized from an oxime (compound of Formula (VIII)). For example, when Xi is Cl, the reaction may be:

[0144] "

[0145]

[0146] In some embodiments, step a) further comprises reacting a compound of Formula (VIII) with N-chlorosuccinimide in order to form compound of Formula (II).

[0147] In some embodiments, this step is performed in situ in the second reactor.

[0148] In some embodiments, a mole ratio of compound of Formula (VIII) to N-chlorosuccinimide is about 1:1 to about 1.2:1. In some embodiments, a mole ratio of compound of Formula (VIII) to N-chlorosuccinimide is about 1.05:1.

[0149] In some embodiments, the reaction is performed at ambient temperature. In some embodiments, the reaction is performed at about 10 °C to about 60 °C, or about 10 °C to about 50 °C. Cooling may be required as the reaction is exothermic.

[0150] In some embodiments, the reaction is performed for a duration of about 1 h to about 5 h, or about 1 h to about 3 h. In some embodiments, the reaction is performed for a duration of about 1 h.

[0151] In some embodiments, the reaction is performed in ethyl acetate. Additional solvent may be added to help regulate the temperature in the reactor.

[0152] The compound of Formula (VIII) (oxime) may be synthesized from starting materials in the second reactor. > >

[0153]

[0154] In some embodiments, the reaction is

[0155] > >

[0156]

[0157] The oxime may be synthesised as either the E- or Z-form, which may equilibrate in solution to an approx. 1:1 mixture. The subsequent reaction to form a compound of Formula (II) involves a chlorination and deprotonation sequence, to give a nitrile oxide which does not have a stereochemical double bond, indicating that the E / Z isomer identity of the oxime does not influence the stereochemistry of later steps.

[0158] In some embodiments, the method further comprises a step before step a) of synthesising the compound of Formula (VIII) in the second reactor. For example, a suitable aldehyde (such as 3-phenylpropionaldehyde) and hydroxylamine hydrochloride may be reacted. In some embodiments, the method further comprises flowing an aldehyde into a solution comprising hydroxylamine hydrochloride and a base. The base may be sodium hydroxide. The aldehyde may be 3-phenylpropionaldehyde.

[0159] In some embodiments, a mole ratio of aldehyde to hydroxylamine hydrochloride is about 0.7:1 to about 1:1.

[0160] In some embodiments, the reaction is performed in an aqueous medium. The aqueous medium may be aqueous methanol. The volume ratio of water to methanol may be about 1:1 to about 1:4, or about 1:2 to about 1:3. In some embodiments, the volume ratio is about 1:2.5.

[0161] In some embodiments, the reaction is performed at ambient temperature. In some embodiments, the reaction is performed at about 10 °C to about 60 °C, or about 10 °C to about 50 °C. Cooling may be required as the reaction is exothermic.

[0162] In some embodiments, the reaction is performed for a duration of about 1 h to about 5 h. In some embodiments, the reaction is performed for a duration of about 2 h.In some embodiments, the reaction is processed by distilling the solvent in order to precipitate the compound of Formula (VIII).

[0163] In some embodiments, the compound of Formula (VIII) is filtered. This may be performed in a filtration means, such as a filter press.

[0164] In some embodiments, the compound of Formula (VIII) is washed. The washing may be performed in a non-solvent. The non-solvent may be a polar solvent. For example, an aqueous medium and / or ethyl acetate may be used.

[0165] In some embodiments, the compound of Formula (VIII) is dried. This may be performed in a drying means, such as a drying oven. The drying may be performed at a temperature of about 50 °C to about 200 °C.

[0166] In some embodiments, the step of step of synthesising the compound of Formula (V) in the first reactor is concurrent with the step of synthesising the compound of Formula (VIII) in the second reactor.

[0167] In some embodiments, the second reactor is characterized by a volume of at least two times the volume of the first reactor. In some embodiments, the second reactor is characterized by a volume of at least 200L.

[0168] Sulfation

[0169] Step b) of the method involves sulphating the thiohydroximate.

[0170]

[0171] In some embodiments, a mole ratio of compound of Formula (III) to pyridine sulfur trioxide is about 1:1.4 to about 1:2. In some embodiments, the mole ratio is about 1:1.5.

[0172] In some embodiments, step b) further comprises adding pyridine.

[0173] In some embodiments, a mole ratio of compound of Formula (III) to pyridine is about1:0.3 to about 1:0.7. In some embodiments, the mole ratio is about 1:0.5.

[0174] In some embodiments, the reaction is performed at a temperature of about 60 °C to about 90 °C. In some embodiments, the reaction is performed at a temperature of about 80 °C. In some embodiments, the reaction is performed at reflux. This means that the reaction is performed at a temperature at or above the boiling point of the solvent.

[0175] In some embodiments, the solvent is a polar aprotic solvent. In some embodiments, the solvent is acetonitrile.

[0176] In some embodiments, the reaction is performed for a duration of about 0.5 h to about 5 h. In some embodiments, the reaction is performed for a duration of about 1 h.

[0177] In some embodiments, step b) further comprises exchanging a cation associated with compound of Formula (IV). In some embodiments, the cation associated with compound of Formula (IV) is exchanged in the presence of a metal carbonate. For example, potassium carbonate in water may be added and stir until effervescence stops.

[0178] In some embodiments, the compound of Formula (IV) is purified. An aqueous medium may be added and removed, which also removes the carbonate salt and other impurities.

[0179] In some embodiments, step b) further comprises precipitating the compound of Formula (IV). In some embodiments, step b) further comprises crystallising the compound of Formula (IV). In some embodiments, the compound of Formula (IV) is recrystallised.

[0180] In some embodiments, the compound of Formula (IV) is filtered. This may be performed in a filtration means, such as a filter press.

[0181] In some embodiments, the compound of Formula (IV) is washed. The washing may be performed in a non-solvent. The non-solvent may be a polar solvent. For example, an aqueous medium and / or ethyl acetate may be used.

[0182] In some embodiments, the compound of Formula (IV) is dried. This may be performed in a drying means, such as a drying oven. The drying may be performed at a temperature of about 50 °C to about 200 °C.Deprotection

[0183] If the compound of Formula (I) comprises protecting groups, the compound of Formula (IV) may be de-protected. In this regard, the method may further comprise a step after step b) of hydrolysing compound of Formula (IV) such that R2 is H.

[0184]

[0185] For example, if the protecting moiety is acyl such as acetyl, compound of Formula (IV) may be deprotected using a base. For example, KOH in methanol may be used.

[0186] The final product may be further purified using resin and / or charcoal. The final product may provided as a solution or as a dry powder.

[0187] In some embodiments, the method further comprises a step of dissolving the compound of Formula (IV) in an aqueous medium. In some embodiments, the method further comprises a step of spray drying the compound of Formula (IV).

[0188] In some embodiments, the method is configured to produce at least 5 kg of compound of Formula (IV).

[0189] In some embodiments, the method is characterised by a duration of about 30 h. This may be optimised by performing reactions concurrently in the first and second reactors.

[0190] As shown in Figure 7, the synthesis of glucosinolate may be completed in five steps. In general, this may occur by synthesizing the glucose isothiourea in the first reactor:

[0191]

[0192] (VI) (VII) (V)Concurrently, the oxime may be synthesised in the second reactor.

[0193] 1 eq. o

[0194] < > "

[0195]

[0196] The glucose thiohydroximate is synthesized by forming the thioglucose in the first reactor and forming the compound of Formula (II) in the second reactor, and then flowing the thioglucose into the second reactor.

[0197] >

[0198]

[0199] The sulphated thiohydroximate is synthesised in the second reactor.

[0200]

[0201] The glucosinolate is produced through deprotection.

[0202]

[0203] In some embodiments, the method of manufacturing glucosinolate, or a derivative, analog, stereoisomer or salt thereof, comprises:

[0204] i) reacting a compound of Formula (V) with sodium sulfite in a first reactor in order to form the compound of Formula (I):

[0205]

[0206] wherein F is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;

[0207] ii) reacting a compound of Formula (VIII) with N-chlorosuccinimide in a second reactor in order to form compound of Formula (II):

[0208]

[0209] iii) flowing a stream comprising a compound of Formula (I) from a first reactor into a solution comprising a compound of Formula (II) in a second reactor in order to form a compound of Formula (III); and

[0210]

[0211] iv) reacting the compound of Formula (III) with pyridine sulfur trioixde in the second reactor in order to form a compound of Formula (IV);

[0212]

[0213] wherein

[0214] Ri is selected from H;

[0215] each R2 is independently selected from H, optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted acyl;

[0216] R3 is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;

[0217] Xi is halo; and

[0218] v) deprotecting the compound of Formula (IV) to form the glucosinolate.

[0219] In some embodiments, step i) further comprises synthesising the compound of Formula (V) in the first reactor by reacting a compound of Formula (VI) with thiourea of Formula (VII):

[0220]

[0221] wherein X2 is halo.

[0222] In some embodiments, step i) and step ii) are performed concurrently.

[0223] In some embodiments, the method further comprises a step after step iv) of exchanging a cation associated with compound of Formula (IV).

[0224] The compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., enantiomers and / or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared byasymmetric syntheses. The invention additionally encompasses compounds described herein as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers. "Optically-enriched," as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments the compound of the present invention is made up of at least about 90% by weight of a preferred stereoisomer. In other embodiments the compound is made up of at least about 95%, 98%, or 99% by weight of a preferred stereoisomer. Preferred stereoisomers may be isolated from racemic mixtures by any method known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts or prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ, of Notre Dame Press, Notre Dame, IN 1972).

[0225] The glucosinolate of the present disclosure has 5 chiral centers and E / Z isomerism at the C=N double bond (as shown below).

[0226]

[0227] The present disclosure also concerns a system for manufacturing glucosinolate, or a derivative, analog, stereoisomer or salt thereof, comprising:

[0228] a) a first reactor;

[0229] b) a second reactor in fluid communication with the first reactor via a first one way valve;

[0230] c) filtration means in fluid communication with the first reactor via a second one way valve and separately in fluid communication with the second reactor via a third one way valve;

[0231] d) drying means in fluid communication with the filtration means and separately in fluid communication with the first and second reactors; and

[0232] e) a spray dryer in fluid communication with the first reactor.

[0233] The first reactor and second reactor are set up such that fluid may only flow from the first reactor to the second reactor. Fluid from the first reactor and second reactor may also only flow into the filtration means via the second and third one way valverespectively.

[0234] As illustrated in Figure 7, in use, the compound of Formula (VIII) (oxime) is synthesised in the second reactor (200L). The compound of Formula (VIII) is passed into the filtration means for purification and then optionally into the drying means. While compound of Formula (VIII) is being synthesized, the compound of Formula (V) (glucose isothiourea) is synthesised in the first reactor (100L). The duration for synthesising the compound of Formula (V) may be modulated or longer such that the use of the filtration means and / or drying means may be sequential. For example, after the compound of Formula (VIII) has been filtered in the filtration means and transferred into the drying means, a mixture comprising compound of Formula (V) may flow into the filtration means to remove impurities and retain compound of Formula (V) as the precipitate. During this time, the compound of Formula (VIII) is flowed back to the second reactor. The compound of Formula (V) may then be optionally dried in the drying means for purification. The duration of these two steps happening concurrently may be about 10 h.

[0235] The compound of Formula (V) (glucose isothiourea) is reacted with sodium sulfite in the first reactor in order to form the compound of Formula (I). Concurrently, the compound of Formula (VIII) is reacted with N-chlorosuccinimide in a second reactor in order to form compound of Formula (II). The compound of Formula (I) dissolved in ethyl acetate is then flowed into the second reactor for reaction with the compound of Formula (II), which is dissolved in ethyl acetate. This reaction solution is then filtered via the filtration means and dried via the drying means to give the compound of Formula (III) (thiohydroximate). The duration of this step may be about 5 h.

[0236] The compound of Formula (III) is reacted with pyridine sulfur trioixde in the second reactor in order to form a compound of Formula (IV) (sulfated thiohydroximate). This may involve flowing the reaction mixture to the filtration means to isolate a crude product, transferring the solid residue back to the second reactor in order to recrystallise the compound of Formula (IV), and flowing the compound of Formula (IV) through the filtration means in order to isolate the product. The compound of Formula (IV) may then be dried in the drying means. The duration of this step may be about 9 h.

[0237] If desired, the compound of Formula (IV) may be deprotected to form the glucosinolate product. Exchange resin may be added to the second reactor, the solution flowed into the filtration means and then into the first reactor for concentration. If desired, the glucosinolate product may be flowed to a spray dryer to obtain the glucosinolate productas a solid.

[0238] Depending on the solvents and impurities, there may be a need to replace the filter.

[0239] Examples

[0240] Laboratory-scale to industrial-scale of glucosinolate (GSL) synthesis

[0241] Combination of block flow diagram (BFD), process flow diagram (PFD) and mass balance calculations have been employed for scale up study of laboratory process. Using the oxime pathway, GSL synthesis was increased from a gram scale to a kilogram scale.

[0242] The overall process of GSL synthesis involves five steps, involving fabrication of oxime, glucose isothiourea, thiohydroximate, sulfated thiohydroximate, and glucosinolate (GSL) (Figure 1). The procedure is suitable for a variety of GSLs, however we used 2-phenylethyl glucosinolate, known also as gluconasturtiin as an exemplary study. It should be highlighted that other GSL derivatives and / or analogs may be synthesized using this process.

[0243] A BFD is constructed for each step of the synthesis. Figure 2 shows step 1, the formation of oxime. Figure 3 shows step 2, the formation of glucose isothiourea. Figure 4 shows step 3, the formation of thiohydroximate. Figure 5 shows step 4, the formation of sulphated thiohydroximate. Figure 6 shows step 5, the formation of glucosinolate.

[0244] Oxime Synthesis:

[0245] • Vacuum distillation of 3-phenylpropionaldehyde (b.p. 222 °C, ca. 107 °C at 20 mmHg) - optional step

[0246] • Prepare sodium hydroxide (11.06 g, 0.277 mol) in aqueous methanol (100 mL water, 250 mL methanol)

[0247] • Add hydroxylamine hydrochloride (20.83 g, 0.300 mol)

[0248] • Add 3-phenylpropionaldehyde (33.58 g, 0.250 mol) (mild exotherm)

[0249] • React for 2 hours (1. Reaction of Figure 2)

[0250] • Distill to remove methanol (ca. 200 grams) - this precipitates the product from solution (2. Distillation of Figure 2)

[0251] • Filter to isolate product (3. Filtration of Figure 2)

[0252] • Wash filter cake with water (2x 25 mL)

[0253] • Dry solid oxime (36.2 g, 97%) - optional step (4. Drying of Figure 2)

[0254] Glucose isothiourea synthesis:• Combine glucose (50.04 g, 0.278 mol), acetic anhydride (140 mL, 1.48 mol) and hydrogen bromide (33% (w / w) in acetic acid, 80 mL, 0.444 mol) - highly exothermic. Initial reaction at r.t. gives mild heating, but accelerates. Use cooling to keep exotherm below 55 °C. (5. Reaction of Figure 3)

[0255] • Once all glucose has dissolved, stir for 3 hours at 35 °C

[0256] • Add sodium acetate (13.64 g, 0.166 mol) to neutralize excess hydrogen bromide • Filter to remove sodium bromide precipitate - intermediate is in solution (6.

[0257] Filtration of Figure 3)

[0258] • Add thiourea (21.12 g, 0.277 mol) to filtrate (7. Reaction of Figure 3)

[0259] • Heat at 80 °C for 1 hour - this precipitates the product from solution. Requires additional solvent to aid in mixing

[0260] • Filter to isolate product (8. Filtration of Figure 3)

[0261] • Wash filter cake with ethyl acetate (80 mL) to give approx. 175 grams wet product (53% solids).

[0262] • Dry solid isothiourea (93.35 g, 68%) - if previous step removes sufficient acetic acid, this is optional (9. Drying of Figure 3)

[0263] Thiohydroximate synthesis:

[0264] First reactor

[0265] • Combine sodium sulfite (12.61 g, 0.100 mol), water (50 mL) and ethyl acetate (50 mL) - more sodium sulfite is required to neutralise any acetic acid remaining in the glucose material (if wet) (10. Reaction of Figure 4)

[0266] • Pass stream of nitrogen or argon through SOLUTION to remove oxygen

[0267] • Add glucose isothiourea (48.86 g, 0.100 mol)

[0268] • React for 1 hour until all solid has dissolved

[0269] • Separate organic fraction (upper layer) (11. Separation of Figure 4)

[0270] Second reactor

[0271] • Combine oxime (15.66 g, 0.105 mol), N-chlorosuccinimide (13.38 g, 0.100 mol), ethyl acetate (100 mL) and HCI (0.4 mL, 0.05 mol) (12. Reaction of Figure 4) • Stir at room temperature for 1 hr - CAUTION - mild exotherm

[0272] • Add extra ethyl acetate (50 mL)

[0273] • Add organic fraction from Vessel A (step S28)

[0274] • Add potassium carbonate (16.63 g, 0.120 mol) in water (40 mL) solution

[0275] • Stir 1 hr - NOTE - this precipitates the product from solution

[0276] • Filter to isolate product (13. Filtration of Figure 4)

[0277] • Wash with water (50 mL) and ethyl acetate (50 mL)• Dry product (37.94 g, 72%, with ca. 2.3% (w / w) succinimide) (14. Drying of Figure 4)

[0278] • Optional: concentrate organic phase to ca. 50 grams, repeat last 3 steps to obtain ca. 10% more product (3.73 g, 7%, with ca. 4.8% (w / w) succinimide)

[0279] Sulfated thiohydroximate synthesis:

[0280] • Combine thiohydroximate (36.51 g, 69.7 mmol, with ca. 2.3% (w / w) succinimide), acetonitrile (35 mL), pyridine (2.8 mL, 35 mmol), pyridine sulfur trioxide (16.63 g, 104 mmol) (15. Reaction of Figure 5)

[0281] • Stir at reflux (ca. 80 °C) for 1 hr

[0282] • Cool to ca 60 °C - NOTE - product may crystallise if allowed to cool further • Add ethyl acetate (35 mL)

[0283] • SLOWLY add potassium carbonate (11.61 g, 84 mmol) in water (35 mL) solution and stir until effervescence stops

[0284] • Add water (35 mL)

[0285] • Remove aqueous phase (16. Separation of Figure 5)

[0286] • Concentrate organic phase to remove ethyl acetate, acetonitrile and crystallise product*

[0287] • Filter to isolate product

[0288] • Wash successively with water (50 mL) and ethyl acetate (50 mL)**

[0289] • Recrystallise product from ethyl acetate and water (17. Distillation & Crystallisation of Figure 5)

[0290] • Filter to isolate product (18. Filtration of Figure 5)

[0291] • Wash successively with water and ethyl acetate

[0292] • Dry product (33.46 g, 76%) (18. Drying of Figure 5)

[0293] * If concentrated to dryness, gives 44.1 grams.

[0294] **If concentrated to dryness after 1st crystallization, gives 36.3 grams.

[0295] Glucosinolate synthesis:

[0296] • Combine sulfated thiohydroximate (16.39 g, 26 mmol), methanol (78 mL) • Add 2 molL'1potassium hydroxide in methanol solution (0.65 mL, 1.3 mmol) (20. Reaction of Figure 6)

[0297] • Stir for 3 hours

[0298] • Add Amberlite MAC-H or other weak acidic resin (carboxylic acid functionality, ca. 1.3 g: 1 gram / mmol potassium hydroxide)

[0299] • Add charcoal to decolourise solution (50 mg), stir for 10 minutes

[0300] • Filter to remove Amberlite MAC-H and charcoal (21. Filtration of Figure 6)Concentrate solution to remove methanol and obtain product (11.76 g, 98%)*

[0301] *Replace this step with solvent swap add water, remove methanol. Provide final product as 50% (w / w) solution in water

[0302] Based on the developed BFDs and the evaluated material flow across different stages, we used two reactors to minimise the process duration to 30 hours per batch. To achieve an annual production capacity of 2000 kg GSL, 250 batches will be required, each producing 8 kg GSL. Mass balance calculations indicated that a jacketed glass reactor should have a capacity of 200 L, while the other glass reactor requires 100 L. The task scheduling outlined in the Gantt Chart (Figure 7) highlights that a total of five pieces of equipment are needed to meet the production timeline.

[0303] A PFD for the GSL manufacturing process was developed as displayed in Figure 8.

[0304] Possible process modifications

[0305] It was found that several aspects of the process may be modified to further improve the process.

[0306] 1. Additional solvent may be used in process steps 7, 8 and 12 (input streams S17, S19 and S30) to aid in solids handling.

[0307] 2. Distillation of the aldehyde before oxime formation is not required, despite use of this technique in a typical lab-scale procedure. Impurities in the aldehyde (94% purity) did not adversely affect the oxime or thiohydroximate reactions, with no significant difference in yield.

[0308] 3. Sulfated thiohydroximate may be directly crystallised using process step 17 (input stream S43), avoiding a lengthy drying process step.

[0309] 4. Sulfated thiohydroximate may not require drying after this crystallisation, with direct recrystallisation to give 2 rounds of purification. Overall this means the isolation of this material using improvements 3. and 4. now requires just one drying cycle, down from three. These changes provide an approximate 25% time savings across the entire process.

[0310] Several process steps may be eliminated from the BFD.

[0311] Process step 4: Use of wet oxime for the subsequent thiohydroximate formation is possible, with no appreciable reduction in product yield. Nevertheless, the preferred process described above retains this drying step, which does not affect the overall process duration or the number of equipment units required.Process step 9: Use of wet glucose isothiourea for the subsequent thiohydroximate formation is possible, however additional quantities of reagents may be required to account for the quantity of acetic acid still present within the wet crystals.

[0312] Process step 15: Use of chlorosulfonic acid as an alternative to expensive pyridine sulfur trioxide provided similar yield for this step (63% vs. 70%).

[0313] Process steps 22 and 23: These involve isolation of pure GSL from a solution in methanol, in which the majority may be removed via distillation, followed by spray drying of a highly concentrated solution. Freeze-drying is typically not compatible with solvents such as methanol, therefore we investigated the use of a solvent swap to provide an aqueous solution of GSL. This may be the final product (provided it is stable), or it may be freeze-dried. Given the larger energy requirements for removing water, the preferred process described above utilises spray dryer, without a water solvent swap or freeze-drying.

[0314] Stability studies:

[0315] Process step 10: The thiosugar obtained from this step is susceptible to dimerization under oxidative conditions. Degassing (specifically, deoxygenation) of the reaction mixture is preferred. However stability studies indicate the likely loss in thiohydroximate yield, due to a thiosugar disulfide byproduct is only up to 3%.

[0316] Process step 12: The activated oxime is prone to rapid dimerization after treatment with potassium carbonate. In the absence of thiosugar (process step 10, output stream S28) studies indicated that approx. 40% of the material had dimerized just five minutes after potassium carbonate addition. Addition of thiosugar solution from process step 10 (input stream S28) should occur BEFORE the addition of input stream S30 to avoid reduced thiohydroximate yield. Further improvements may be expected if the potassium carbonate is added portionwise or over a period of time.

[0317] Thermal hazard testing:

[0318] Process steps 5 and 12 were noted to be exothermic during lab-based studies.

[0319] Process step 5:

[0320] This step exhibited a significant exotherm, which was performed only up to 1 mol scale (ca. 2% of recommended batch size). A temperature below 50 °C is recommended, to avoid degradation of the sugar. Yet the rate of reaction is slow at room temperature, requiring careful temperature control within the range of ca. 30-50 °C by partial cooling with an ice bath. Given the sensitivity of this process, thermal hazard testing was conducted, indicating an exotherm of ca. 10,900 kJ at the chosen manufacturing scale.Process step 12:

[0321] This step exhibited a significant exotherm heating the reaction to ca. 60 °C, when performed at up to 300 mmol (ca. 1% of recommended batch size). Given higher scales would likely lead to boiling of the solvent, thermal hazard testing was conducted, indicating an exotherm of ca. 4,800 kJ at the chosen manufacturing scale.

[0322] Both process steps exhibited significant heat, in which the reaction conditions tested for thermal hazards produce up to 2x more heat than the reactor can remove. Ensuring the reaction temperatures are approx. 20 °C is expected to be suitable for both, with reaction time is expected to only marginally increase.

[0323] It will be appreciated that many further modifications and permutations of various aspects of the described embodiments are possible. Accordingly, the described aspects are intended to embrace all such alterations, modifications, and variations that fall within the spirit and scope of the appended claims.

[0324] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0325] Throughout this specification and the claims which follow, unless the context requires otherwise, the phrase "consisting essentially of", and variations such as "consists essentially of" will be understood to indicate that the recited element(s) is / are essential i.e. necessary elements of the invention. The phrase allows for the presence of other non-recited elements which do not materially affect the characteristics of the invention but excludes additional unspecified elements which would affect the basic and novel characteristics of the method defined.

[0326] The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

Claims

1. Claims1. A method of manufacturing glucosinolate, or a derivative, analog, stereoisomer or salt thereof, comprising:a) flowing a stream comprising a compound of Formula (I) from a first reactor into a solution comprising a compound of Formula (II) in a second reactor in order to form a compound of Formula (III); andb) reacting the compound of Formula (III) with pyridine sulfur trioixde in the second reactor in order to form a compound of Formula (IV);whereinRi is selected from H;each R2 is independently selected from H, optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted acyl;Rs is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; andXi is halo.

2. The method according to claim 1, wherein a mole ratio of compound of Formula (I) to compound of Formula (II) is about 1:1.02 to about 1:1.1.

3. The method according to claim 1 or 2, wherein step a) further comprises adding ethyl acetate to the second reactor before flowing the stream comprising a compound of Formula (I) into the second reactor.

4. The method according to any one of claims 1 to 3, wherein step a) further comprises adding a carbonate to the second reactor in order to precipitate the compound of Formula (III).

5. The method according to any one of claims 1 to 4, wherein the compound of Formula (III) is concentrated to less than about 0.5 times its initial volume.

6. The method according to any one of claims 1 to 5, wherein the compound of Formula (III) is the compound of Formula (III) is filtered, washed, and / or dried.

7. The method according to any one of claims 1 to 6, wherein step a) further comprises reacting a compound of Formula (V) with sodium sulfite (sulphite) in the first reactor in order to form the compound of Formula (I):wherein F is selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl.

8. The method according to claim 7, wherein a mole ratio of compound of Formula (V) and sodium sulfite is about 1:1 to about 1:1.5.

9. The method according to claim 7 or 8, wherein the reaction is performed for a duration of at least 1 h.

10. The method according to any one of claims 7 to 9, wherein reaction is performed in a biphasic mixture of water and ethyl acetate.

11. The method according to 10, wherein the ethyl acetate phase is flowed into the second reactor.

12. The method according to any one of claims 7 to 11, wherein the method further comprises a step before step a) of synthesising the compound of Formula (V) in the first reactor.

13. The method according to claim 12, wherein the compound of Formula (V) is synthesised by reacting a compound of Formula (VI) with thiourea of Formula (VII):wherein X2 is halo.

14. The method according to claim 12 or 13, wherein a mole ratio of compound of Formula (VI) to thiourea of Formula (VII) is about 1:1 to about 1:2.

15. The method according to any one of claims 12 to 14, wherein the reaction is performed at a temperature of about 60 °C to about 90 °C for a duration of about 0.5 h to about 5 h in order to precipitate compound of Formula (V).

16. The method according to any one of claims 12 to 15, wherein the compound of Formula (V) is filtered, washed and / or dried.

17. The method according to any one of claims 1 to 16, wherein the first reactor is characterized by a volume of at least 100L.

18. The method according to any one of claims 1 to 17, wherein step a) further comprises reacting a compound of Formula (VIII) with N-chlorosuccinimide in order to form compound of Formula (II);19. The method according to claim 18, wherein the reaction is performed at ambient temperature for a duration of about 1 h to about 5 h.

20. The method according to claim 18 or 19, wherein the reaction is performed in ethyl acetate.

21. The method according to any one of claims 18 to 20, wherein the method further comprises a step before step a) of synthesising the compound of Formula (VIII) in the second reactor.

22. The method according to claim 21, wherein the reaction is performed at ambient temperature for a duration of about 1 h to about 5 h.

23. The method according to claim 21 or 22, wherein the reaction is distilled in order to precipitate the compound of Formula (VIII).

24. The method according to any one of claims 21 to 23, wherein the compound of Formula (VIII) is filtered, washed and / or dried.

25. The method according to any one of claims 13 to 24, wherein the step of synthesising the compound of Formula (V) in the first reactor is concurrent with the step of synthesising the compound of Formula (VIII) in the second reactor.

26. The method according to any one of claims 1 to 25, wherein the second reactor is characterized by a volume of at least two times the volume of the first reactor.

27. The method according to any one of claims 1 to 26, wherein a mole ratio of compound of Formula (III) to pyridine sulfur trioxide is about 1:1.4 to about 1:2.

28. The method according to any one of claims 1 to 27, wherein step b) further comprises adding pyridine.

29. The method according to claim 28, wherein a mole ratio of compound of Formula (III) to pyridine is about 1:0.3 to about 1:0.7.

30. The method according to any one of claims 1 to 29, wherein step b) is performed at a temperature of about 60 °C to about 90 °C for a duration of about 0.5 h to about 5 h.

31. The method according to any one of claims 1 to 30, wherein step b) further comprises exchanging a cation associated with compound of Formula (IV).

32. The method according to any one of claims 1 to 31, wherein step b) further comprises precipitating the compound of Formula (IV).

33. The method according to any one of claims 1 to 32, wherein the compound of Formula (IV) is filtered, washed, dried and / or recrystallized.

34. The method according to any one of claims 1 to 33, wherein the method further comprises a step after step b) of hydrolysing compound of Formula (IV) such that F is H.

35. The method according to any one of claims 1 to 34, wherein the method is configured to produce at least 5 kg of compound of Formula (IV).

36. The method according to any one of claims 1 to 35, wherein the method is characterised by a duration of about 30 h.

37. A system for manufacturing glucosinolate, or a derivative, analog, stereoisomer or salt thereof based on the method according to any one of claims 1 to 36, comprising: a) a first reactor;b) a second reactor in fluid communication with the first reactor via a first one way valve;c) filtration means in fluid communication with the first reactor via a second one way valve and separately in fluid communication with the second reactor via a third one way valve;d) drying means in fluid communication with the filtration means and separately in fluid communication with the first and second reactors; ande) a spray dryer in fluid communication with the first reactor.

Images