[0001] The present disclosure relates to a method of preparing heteropolycyclic compounds such as substituted 1,2,3,4-tetrahydroisoquinoline compounds using biotransformation reactions. In particular embodiments, the heteropolycyclic compounds are prepared by Pictet-Spengler reaction using a Pictet-Spenglerase biocatalyst and the aldehyde substrate for the Pictet-Spenglerase biocatalyst reaction is produced in situ by a transaminase biocatalyst. The use of the heteropolycyclic compounds is also described.

Background of the Disclosure

[0002] The Pictet-Spengler reaction comprises the condensation of an arylalkylamine and a carbonyl containing compound, followed by an intramolecular cyclisation to produce a 1,2,3,4-tetrahydroisoquinoline compound. This reaction is an uncommon reaction for a biocatalyst to facilitate, but at least one biocatalyst, Norcoclaurine Synthase (NCS, EC.4.2.1.78) catalyses the Pictet-Spengler reaction between dopamine and

hydroxyphenylacetaldehyde to produce norcoclaurine and water. Norcoclaurine is an important intermediate in the synthesis of benzylisoquinoline alkaloids. While it has been shown that NCS can accept a wider variety of substrates, promiscuously catalysing the reaction between a number of aldehydes and amines to generate new products, the aldehydes used are either purchased or synthesized chemically prior to use.

[0003] Chemical synthesis of the aldehydes can require harsh chemical treatments, extractions and purifications. Often the aldehydes are unstable and as the complexity of the aldehyde increases, the low selectivity of chemical approaches and instability of the aldehydes leads to mixed products and low yields.

[0004] There is a need for a method of producing heteropolycyclic compounds such as tetrahydroisoquinoline compounds, utilizing aldehydes of varying complexity and that are produced in situ for immediate use in the Pictet-Spengler reaction. Summary of the Disclosure

[0005] The present disclosure is predicated at least in part by the discovery that transaminase biocatalysts can be used to produce aldehydes that can be further utilized in a Pictet-Spengler biotransformation reaction in situ, under ambient conditions and in an aqueous environment. The "one pot" method allows the production of aldehydes, including unstable aldehydes, that are rapidly used in the biocatalysed Pictet-Spengler reaction to produce 1,2,3,4-tetrahydroisoquinoline compounds under mild conditions and in good yield.

[0006] In a first aspect of the present disclosure, there is provided a method of preparing a l-substituted-2N-containing fused heteropolycyclic compound comprising: i) contacting an optionally substituted alkylamine compound with a transaminase biocatalyst in the presence of a carbonyl-containing co- substrate to provide an optionally substituted alkylaldehyde compound; and

ii) contacting the optionally substituted alkylaldehyde compound with a Pictet-Spenglerase biocatalyst in the presence of an optionally substituted heteroaryl or aryl alkylamine compound.

[0007] In another aspect of the disclosure, there is provided a use of a compound of formula (VII):

wherein Ar is an optionally substituted aryl group further substituted with an electron pair donor in a position β to the ring junction carbon atom labeled 4, or an optionally substituted heteroaryl group; Ri is selected from -CH ₃ and an optionally substituted aryl or heteroaryl group;

Each R ₂ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ;

Each R ₃ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl and heteroaryl;

Each R ₄ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl and -OR ₃ ;

Each R5 is independently selected from hydrogen and halogen;

Each R ₆ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ ,

-OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and

-OC(R ₅ ) ₃ ;

R ₇ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, -OH, -Oalkyl, -Oalkenyl and -Oalkynyl;

n is an integer from 1 to 6; and

p is an integer from 0 to 12;

as an intermediate in the synthesis of a biologically active compound.

[0008] In another aspect of the disclosure, there is provided an optionally substituted l-substituted-2N-containing fused heteropolycyclic compound prepared by the method of the disclosure.

[0009] In particular embodiments, the l-substituted-2N-containing fused

heteropolycyclic compound is a 1 -alkyl- or 1-arylalkyl- or l-heteroarylalkyl-1,2,3,4- tetrahydroisoquinoline compound.

[0010] In yet another aspect of the disclosure there is provided a compound of formula (X):

wherein Ar is an optionally substituted aryl or heteroaryl group; r is 0 or an integer from 1 to 4;

Each of R ₂ o, R ₂₂ and R ₂₃ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₂₈ , -CO2R28, -C(O)R ₂₈ , -alkyl-OH, -C(O)N(R ₂₈ ) ₂ , -N(R ₂₉ ) ₂ , -C(R ₃₀ ) ₃ and -OC(R ₃₀ ) ₃ ;

R ₂₁ is an electron pair donor;

Each of R ₂₄ and R ₂₅ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₂₈ , -CO ₂ R ₂₈ , -C(O)R ₂₈ , -alkyl-OH, -C(O)N(R ₂₈ ) ₂ , -N(R ₂₉ ) ₂ , -C(R ₃₀ ) and -OC(R ₃₀ ) ₃ ;

R ₂₆ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, -OH, -Oalkyl, Oalkenyl and -Oalkynyl;

Each R ₂₇ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₂₈ , -CO ₂ R _2S ,

-C(O)R ₂₈ , -alkyl-OH, -C(O)N(R ₂₈ ) ₂ , -N(R ₂₉ ) ₂ , -C(R ₃₀ ) ₃ and -OC(R ₃₀ ) ₃ ;

Each R ₂₈ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl and heteroaryl;

Each R ₂ 9 is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl and -OR ₂₈ ;

Each R ₃ o is independently selected from hydrogen and halogen; with the proviso that: when R ₂₀ , R23, R24, R25, R26 and R ₂₇ are hydrogen, R ₂ i and R ₂₂ are -OH and r is 1, Ar is not unsubstituted phenyl, 4-hydroxyphenyl, 3,4-dihydroxyphenyl or 3-thiophenyl.

Detailed Description of the Disclosure

Definitions

[0011] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, preferred methods and materials are described. For the purposes of the present disclosure, the following terms are defined below.

[0012] The articles "a" and "an" are used herein to refer to one or to more than one (i.e. to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0013] As used herein, the term "about" refers to a quantity, level, value, dimension, size, or amount that varies by as much as 30%, 25%, 20%, 15% or 10% to a reference quantity, level, value, dimension, size, or amount.

[0014] Throughout this specification, unless the context requires otherwise, the words "comprise", "comprises" and "comprising" will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.

[0015] 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 disclosure. The phrase allows for the presence of other non- recited elements which do not materially affect the characteristics of the disclosure but excludes additional unspecified elements which would affect the basic and novel characteristics of the disclosure. [0016] The term "invention" includes all disclosures, aspects, embodiments and examples described herein.

[0017] As used herein, the term "alkyl" refers to a straight chain or branched saturated hydrocarbon group having 1 to 10 carbon atoms. Where appropriate, the alkyl group may have a specified number of carbon atoms, for example, Ci_ ₆ alkyl which includes alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms in a linear or branched arrangement.

Examples of suitable alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, z ^' -butyl, i-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 4-methylbutyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 5-methylpentyl, 2-ethylbutyl, 3-ethylbutyl, heptyl, octyl, nonyl and decyl.

[0018] As used herein, the term "alkenyl" refers to a straight-chain or branched hydrocarbon group having one or more double bonds between carbon atoms and having 2 to 10 carbon atoms. Where appropriate, the alkenyl group may have a specified number of carbon atoms. For example, C ₂ -C ₆ as in "C ₂ -C ₆ alkenyl" includes groups having 2, 3, 4, 5 or 6 carbon atoms in a linear or branched arrangement. Examples of suitable alkenyl groups include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, butadienyl, pentenyl, pentadienyl, hexenyl, hexadienyl, heptenyl, octenyl, nonenyl and decenyl.

[0019] As used herein, the term "alkynyl" refers to a straight-chain or branched hydrocarbon group having one or more triple bonds and having 2 to 10 carbon atoms. Where appropriate, the alkynyl group may have a specified number of carbon atoms. For example, C ₂ -C ₆ as in "C ₂ -C ₆ alkynyl" includes groups having 2, 3, 4, 5 or 6 carbon atoms in a linear or branched arrangement. Examples of suitable alkynyl groups include, but are not limited to ethynyl, propynyl, butynyl, pentynyl and hexynyl.

[0020] As used herein, the term "cycloalkyl" refers to a saturated cyclic hydrocarbon. The cycloalkyl ring may include a specified number of carbon atoms. For example, a 3 to 8 membered cycloalkyl group includes 3, 4, 5, 6, 7 or 8 carbon atoms. Examples of suitable cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. [0021] As used herein, the term "cycloalkenyl" refers to an unsaturated cyclic hydrocarbon. The cycloalkenyl ring may include a specified number of carbon atoms. For example, a 5 to 8 membered cycloalkenyl group includes 5, 6, 7 or 8 carbon atoms. The cycloalkenyl group has one or more double bonds and when more than one double bond is present, the double bonds may be unconjugated or conjugated, however the cycloalkenyl group is not aromatic. Examples of suitable cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, cycloheptatrienyl, cyclooctenyl, cyclooctadienyl and cyclooctatrienyl rings.

[0022] The term "heterocyclic" or "heterocyclyl" as used herein, refers to a cyclic hydrocarbon in which one to four carbon atoms have been replaced by heteroatoms independently selected from the group consisting of N, N(R), S, S(O), S(O) ₂ and O. A heterocyclic ring may be saturated or unsaturated. Examples of suitable heterocyclyl groups include azetidine, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, 2- oxopyrrolidinyl, pyrrolinyl, pyranyl, dioxolanyl, piperidinyl, 2-oxopiperidinyl, pyrazolinyl, imidazolinyl, thiazolinyl, dithiolyl, oxathiolyl, dioxanyl, dioxinyl, dioxazolyl, oxathiozolyl, oxazolonyl, piperazinyl, morpholino, thiomorpholinyl, 3-oxomorpholinyl, dithianyl, trithianyl and oxazinyl.

[0023] As used herein, the term "aryl" is intended to mean any stable, monocyclic, bicyclic or tricyclic carbon ring system of up to 7 atoms in each ring, wherein at least one ring is aromatic. Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, fluorenyl, phenanthrenyl, biphenyl and binaphthyl.

[0024] The term "heteroaryl" as used herein, represents a stable monocyclic, bicyclic or tricyclic ring of up to 7 atoms in each ring, wherein at least one ring is aromatic and at least one ring contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S. Heteroaryl groups within the scope of this definition include, but are not limited to, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, quinazolinyl, pyrazolyl, indolyl, isoindolyl, lH,3H-l-oxoisoindolyl, benzotriazolyl, furanyl, thienyl, thiophenyl, benzothienyl, benzofuranyl, benzodioxane, benzodioxin, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, imidazolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinolinyl, thiazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,5-triazinyl, 1,2,4-triazinyl, 1,2,4,5-tetrazinyl and tetrazolyl. Particular heteroaryl groups have 5- or 6-membered rings, such as pyrazolyl, furanyl, thienyl, oxazolyl, indolyl, isoindolyl, lH,3H-l-oxoisoindolyl, isoxazolyl, imidazolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, thiazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl and 1,2,4-oxadiazolyl and 1,2,4-thiadiazolyl.

[0025] The term "heteropolycyclic" as used herein, refers to a heterocyclic or heteroaryl group that has more than one ring and where the rings are fused to form a fused polycyclic ring system. The heteropolycyclic compound may have one or more rings which are aromatic, unsaturated or saturated or mixtures thereof.

[0026] As used herein, the term "halogen" or "halo" refers to fluorine (fluoro), chlorine (chloro), bromine (bromo) and iodine (iodo).

[0027] Each alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl heterocyclyl and heteroaryl whether an individual entity or as part of a larger entity may be optionally substituted with one or more optional substituents selected from the group consisting of Ci_ ₆ alkyl, oxo (=0), -OH, -SH, Ci_ ₆ alkylO-, Ci_ ₆ alkylS-, -CO ₂ H, -CO ₂ Ci_ ₆ alkyl, -CONH ₂ , -CONH(alkyl), -CON(alkyl) ₂ , -Ci_ ₆ alkylOH, -NH ₂ , -NH(Ci_ ₆ alkyl), -N(Ci_ ₆ alkyl) ₂ , -NH(phenyl), -N(phenyl) ₂ , -CN, -NO ₂ , -halogen, -CF ₃ , -OCF ₃ , -SCF ₃ , -CHF ₂ , -OCHF ₂ , -SCHF ₂ , -phenyl, benzyl, -Ophenyl, -C(O)phenyl and -C(O)Ci_ ₆ alkyl. Examples of suitable substituents include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, vinyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, methylthio, ethylthio, propylthio, isopropylthio, butylthio, hydroxy, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, fluoro, chloro, bromo, iodo, cyano, nitro, -CO ₂ H, -CO ₂ CH ₃ , trifluoromethyl, trifluoromethoxy, trifluoromethylthio, difluoromethyl, difluoromethoxy, difluoromethylthio, amino, methylamino, dimethylamino, phenyl, phenoxy,

phenylcarbonyl, benzyl and acetyl.

[0028] The term "electron pair donor" as used herein refers to an atom or group of atoms that is able to donate a pair of electrons to the carbon atom to which it is attached. Suitable electron pair donors include substituents including an oxygen or nitrogen atom that is able to donate the electron pair to the carbon atom to which it is attached. Suitable electron pair donor groups include, but are not limited to, -OH, -Oalkyl, -Oalkenyl, -Oalkynyl, -Ocycloalkyl, -Ocycloalkenyl, -Oaryl, -Oheterocyclyl, -Oheteroaryl, -OCF ₃ , -NH ₂ , -NH(alkyl), -NH(aryl), -NH(alkenyl), -NH(alkynyl), -NH(cycloalkyl),

-NH(cycloalkenyl), -NH(heterocyclyl), -NH(heteroaryl), -N(alkyl) ₂ , -N(aryl) ₂ ,

-N(alkenyl) ₂ , -N(alkynyl) ₂ , -N(cycloalkyl) ₂ , -N(cycloalkenyl) ₂ , -N(heteroaryl) ₂ ,

-N(heterocyclyl) ₂ , -N(alkyl)(aryl), -N(alkenyl)(aryl), -N(alkynyl)(aryl), -N(alkyl)(alkenyl), -N(alkyl)(alkynyl), -N(cycloalkyl)(aryl), -N(cycloalkenyl)(aryl), -N(alkyl)(heteroaryl), and the like.

[0029] It will also be recognised that compounds produced by the methods of the disclosure possess one or more asymmetric centres and are therefore capable of existing in more than one stereoisomeric form. The disclosure thus also relates to compounds in substantially pure isomeric form at one or more asymmetric centres eg., greater than about 90% ee, such as about 95% or 97% ee or greater than 99% ee, as well as mixtures, including racemic mixtures, thereof. Such isomers may be prepared by asymmetric synthesis, for example using chiral intermediates, biotransformation reactions or by chiral resolution.

[0030] As used herein the term "transaminase biocatalyst" refers to a biocatalyst that catalyses a transamination reaction in which an amino group on one compound is exchanged with a carbonyl group on another compound. The compound that transfers the amino group may have the amino group attached to a primary or secondary carbon atom. The compound that transfers the carbonyl group may be an aldehyde or a ketone, for example, the compound may be a ketocarboxylic acid (keto acid). When the compound that transfers the amino group has the amino group attached to a primary carbon atom, the compound produced is an aldehyde. When the compound that transfers the amino group has the amino group attached to a secondary carbon atom, the compound produced is a ketone. For example, if the amine group is attached to a primary carbon atom and the carbonyl group is part of an a-keto acid, the biotransformation results in an aldehyde and an a- amino acid as shown below.

[0031] As used herein the term "Pictet-Spenglerase biocatalyst" refers to a biocatalyst that catalyses a Pictet-Spengler reaction between a heteroaryl or aryl alkylamine compound and a carbonyl containing compound such as an aldehyde or a ketone, to provide an imine intermediate that undergoes ring closure to produce a nitrogen containing ring fused to an aryl or heteroaryl group. For example, when the heteroaryl or aryl alkylamine is a substituted or unsubstituted phenylethylamine and the carbonyl containing compound is an aldehyde, the product produced by the Pictet-Spenglerase biocatalyst is a substituted or unsubstituted 1,2,3,4-tetrahydroisoquinoline, such as shown below.

Biotransformation Reactions

[0032] In a first aspect of the present disclosure, there is provided a method of preparing a l-substituted-2N-containing fused heteropolycyclic compound comprising: i) contacting an optionally substituted alkylamine compound with a

transaminase biocatalyst in the presence of a carbonyl-containing co- substrate to provide an optionally substituted alkylaldehyde compound; and

ii) contacting the optionally substituted alkylaldehyde compound with a

Pictet-Spenglerase biocatalyst in the presence of an optionally substituted heteroaryl or aryl alkylamine compound. [0033] While it is possible to carry out the method of the disclosure in two separate steps, isolating the aldehyde for use in the second step, in some embodiments it is convenient for both steps of the method to be carried out in a single reaction vessel containing all components of the disclosure simultaneously.

[0034] The optionally substituted alkylamine compound may be any alkylamine compound that has an amine group bound to a primary carbon atom and that can be converted to an aldehyde. In some embodiments, the alkylamine compound is an unsubstituted alkylamine or an alkylamine in which the alkyl group of the alkylamine is substituted with an aryl or heteroaryl group. In some embodiments, the optionally substituted alkylamine is a compound of formula (I):

R ₁ -(CHR ₂ ) _m -CH ₂ -NH ₂ (I) wherein Ri is selected from -CH ₃ and an optionally substituted aryl or heteroaryl group; each R ₂ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and -OC(R ₅ ) ;

Each R ₃ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl and heterocyclyl;

Each R ₄ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heterocyclyl and -OR ₃ ;

Each R5 is independently selected from hydrogen and halogen; and

m is an integer from 0 to 12, such as 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

[0035] The transaminase reaction produces an optionally substituted alkylaldehyde compound and in some embodiments, the optionally substituted alkylaldehyde compound is a compound of formula (II):

R ₁ -(CHR ₂ ) _m -C(O)-H (II) wherein Ri, R ₂ and m are as defined for formula (I).

[0036] In some particular embodiments of formula (I) and (II), one or more of the following applies: Ri is selected from -CH ₃ , phenyl, naphthyl, tetrahydronaphthyl, pyridinyl, pyrimidinyl, pyrazinyl, pyrrolyl, pyrazolyl, imidazolyl, indolyl, benzofuranyl, benzothiophenyl, indolinyl, isoindolyl, benzimidazolyl, quinalinyl, isoquinolinyl, quinazolinyl, quinoxalinyl and naphthyridinyl, each of which may be optionally substituted, especially optionally substituted phenyl, naphthyl, pyridinyl, quinolinyl and isoquinolinyl;

each R ₂ is independently selected from hydrogen, alkyl, halogen, -OH, -Oalkyl, -CO ₂ H, -CO ₂ alkyl, -alkyl-OH, -C(O)N(R ₃ ) ₂ , -NH ₂ , -CF ₃ and -OCF ₃ , especially hydrogen, hydroxyl or -CO ₂ H, more especially hydrogen or hydroxyl;

m is an integer from 1 to 10, especially 1 to 8 or 1 to 6, more especially 1 to 4, most especially 1 or 2.

[0037] In some particular further embodiments, the aryl or heteroaryl substituted alkylamine is selected from 3-amino-l-phenyl-propanol, 3-hydroxy-4-methoxy- phenylethylamine, 6-hydroxydopamine, 3-methoxytyramine, phenylethylamine, 3-phenyl- 1 -propylamine, tryptamine, dopamine, serotonin, octapamine, tyramine, norepinephrine, 2- (l-naphthyl)ethylamine and 3-(l-naphthyl)propylamine.

[0038] The transaminase biocatalyst may be any biocatalyst that performs a transaminase reaction and may be selected to accept specific alkylamine substates, for example a specific aryl or heteroaryl alkylamine substrate to provide a desired aldehyde. In some embodiments, including but not limited to any of the embodiments described above in paragraphs [0034], [0035], [0036] and [0037], the transaminase biocatalyst is a transaminase biocatalyst from a Pseudomonas spp. described in AU2013902128. In some embodiments, the transaminase biocatalyst is a polypeptide (i) of any one of SEQ ID NOs: l to 18; or (ii) an amino acid sequence which is at least 25% identical to any one or more of SEQ ID NOs: 1 to 18; or a biologically active fragment of (i) or (ii).

SEQ ID NO : 1 :

[0039] In some embodiments, the polypeptide is a polypeptide of any one of SEQ ID NOs: 1 to 18. In other embodiments, the polypeptide has at least 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90% or 95% sequence identity with one or more of SEQ ID NOs: 1 to 11, provided the polypeptide has activity as a transaminase biocatalyst.

[0040] In particular embodiments, the transaminase biocatalyst is a substantially purified and/or recombinant polypeptide having any one of the amino acid sequences of SEQ ID NOs: 1 to 4, especially one of SEQ ID NOs: 1 or 4.

[0041] The carbonyl containing co-substrate is any carbonyl containing compound that is able to exchange its carbonyl group for an amino group in the biocatalysed transamination reaction. In some embodiments, the transamination reaction produces an aldehyde together with an amine that are both substrates for the Pictet-Spenglerase biocatalyst. Without wishing to be bound by theory, it is possible that the Pictet- Spenglerase biocatalyst prefers one pair of substrates over a second possible pair of substrates. For example:

While compounds 1 and 2 appear to be substrates for the Pictet-Spenglerase biocatalyst, the reaction does not proceed in the absence of the transaminase biocatalyst suggesting that compounds 3 and 4 are the substrates for the Pictet-Spenglerase biocatalyst.

[0042] In some embodiments, a co-substrate is added to the reaction and the co- substrate and the products produced from the co-substrate in the transaminase reaction are not substrates for the Pictet-Spenglerase biocatalyst.

[0043] In some embodiments, including any of those described in paragraphs [0034], [0035], [0036], [0037] and [0038], the co-substrate is a ketone or aldehyde or a keto acid, especially a ketone or keto acid. In some embodiments, the keto acid may be an a-keto acid, a β-keto acid or a γ-keto acid, especially an a-keto acid. Suitable co-substrates include, but are not limited to, pyruvate, acetone, oxaloacetic acid, a-ketoglutarate, glyoxylic acid, acetoacetic acid and levulinic acid, especially pyruvate, acetone, oxaloacetic acid, glyoxylic acid and a-ketoglutarate, more especially pyruvate and acetone. In these embodiments, the co-substrate produces a by-product in the transaminase reaction. For example, when the co-substrate is a keto-acid, the by-product produced is an amino acid. A particular example is pyruvate as the co-substrate, which produces alanine as a byproduct.

[0044] The Pictet-Spenglerase biocatalyst is any biocatalyst that catalyses a Pictet- Spengler reaction. Examples of Pictet-Spenglerase biocatalysts include Norcoclaurine Synthase (NCS, EC 4.2.1.78) and Strictosidine Synthase (STR, EC 4.3.3.2). Different isoforms of these enzymes may be derived from different plant species. For example, different isoforms of Strictosidine Synthase are found in Catharanthus roseus, Rauvolfia serpentine, Rauvolfia mannii, Rauvolfia verticillata, Ophiorhiza pumila and Ophiorhiza japonica. Different isoforms of Norcoclaurine Synthase are derived from Thalictrum flavum and Coptis japonica. These biocatalysts may be obtained recombinantly. In particular embodiments, the Pictet-Spenglerase biocatalyst is Norcoclaurine Synthase, especially NCS having the polypeptide sequence SEQ ID NO: 12:

MQKLILTGRPFLHHQGIINQVSTVTKVIHHELEVAASADDIWTVYSWPGLAKHLPD LLPGAFEKLEIIGDGGVGTILDMTFVPGEFPHEYKEKFILVDNEHRLKKVQMIEGG YLDLGVTYYMDTIHVVPTGKDSCVIKSSTEYHVKPEFVKIVEPLITTGPLAAMADA IS KLVLEHKS KS NS DEIE A AIIT VLE

[0045] The optionally substituted heteroaryl or aryl alkylamine is any heteroaryl or aryl alkylamine that is a substrate for the Pictet-Spenglerase biocatalyst.

[0046] In some embodiments, the optionally substituted heteroaryl or aryl alkylamine is a compound of formula (III):

Ar-(CHR ₆ ) _n -NH-R ₇ (III)

wherein Ar is an optionally substituted aryl group futher substituted with an electron pair donor in a position β to the attachment to the alkylene chain -(CHR ₆ )n-, or an optionally substituted heteroaryl group; each R ₆ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , - C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and -OC(R ₅ ) ;

[0047] R ₇ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OH, -Oalkyl, -Oalkenyl and -Oalkynyl; n is an integer from 1 to 6, such as 1, 2, 3, 4, 5, or 6; and

R ₃ , R4 and R5 are as defined for formula (I).

[0048] In some embodiments, Ar is an optionally substituted heteroaryl group in which at least one optional substituent is an electron pair donor.

[0049] In some embodiments, including any of those described in paragraphs [0034], [0035], [0036], [0037], [0038] and [0043], the heteroaryl or aryl alkylamine is a compound of formula (IV):

wherein each R ₃ , R ₄ , R5, R ₆ and R ₇ and n are as defined for formula (III); one of V, W, X, Y and Z is -0-, -S- or -NH- and the others are selected from -C= and - N=, or each of V, W, X, Y and Z are independently selected from -C= and -N=; q is 0 or 1, one of R ₈ and R ₁₂ is hydrogen and the other is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , - OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; [0050] Each R9 and R ₁₁ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR3, -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or when all of V, W, X, Y and Z are -C=, one of R9 and R ₁₁ is an electron pair donor and the other is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; R ₁₀ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, - C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or R ₁₀ together with one or R ₉ and R ₁₁ and the atoms to which R ₁₀ and R9 or R ₁₁ are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; or

R ₈ and R9 together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; provided that R ₁₂ is hydrogen; or R ₁₁ and R ₁₂ together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; provided that R ₈ is hydrogen; provided that:

(i) when any of V, W, X, Y or Z is -O-, -S-, -NH- or -N=, the respective substituent R ₁₂ , R ₁₁ , R ₁₀ , R ₉ or R ₈ is absent; and

(ii) when R ₈ is hydrogen, Z is -C= or when R ₁₂ is hydrogen, V is -C=.

[0051] In yet other embodiments, the compound of formula (IV) is an aryl compound of formula (V):

wherein each R ₆ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , - CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ;

R ₇ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OH, -Oalkyl, -Oalkenyl and -Oalkynyl; one of R ₈ and R ₁₂ is hydrogen and the other is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , - OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; one of R9 and R ₁₁ is an electron pair donor and the other is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ;

R ₁₀ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, - C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or R ₁₀ together with one or R9 and R ₁₁ , whichever is not the electron pair donor, and the atoms to which R ₁₀ and R9 or R ₁₁ are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; or

R ₈ and R9 together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; provided that R ₁₂ is hydrogen and R ₁₁ is the electron pair donor; or R ₁₁ and R ₁₂ together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; provided that R ₈ is hydrogen and R9 is the electron pair donor; n is an integer of 1 to 6, such as 1, 2, 3, 4,5, or 6; and

R ₃ , R ₄ and R5 are as defined for formula (I).

[0052] In other embodiments, the heteroaryl or aryl alkylamine compound is a heteroaryl compound of formula (VI):

wherein one of V, W, X, Y and Z is selected from -0-, -S-, -NH- and -N= and the others are selected from -C= and -N=; q is 0 or 1, one of R ₈ and R ₁₂ is hydrogen and the other is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , - OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ;

Each of R9, R ₁₀ and R ₁₁ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , - CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or R ₁₀ together with one or R9 and R ₁₁ and the atoms to which R ₁₀ and R9 or R ₁₁ are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; or R ₈ and R9 together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; provided that R ₁₂ is hydrogen; or

R ₁₁ and R ₁₂ together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; provided that R ₈ is hydrogen;

R ₆ , R7 and n are as defined for formula (III) and R ₃ , R4 and R5 are as defined for formula

(I);

provided that:

(i) when any of V, W, X, Y or Z is -0-, -S-, -NH- or -N=, the respective substituent R ₁₂ , R ₁₁ , R ₁₀ , R9 or R ₈ is absent; and

(ii) when R ₈ is hydrogen, Z is -C= or when R ₁₂ is hydrogen, V is -C=.

[0053] In some embodiments of Formula (III), (IV), (V) and (VI), one or more of the following applies:

Each R ₆ is independently selected from hydrogen, alkyl, -OH, -Oalkyl, -CO ₂ H, -CO ₂ alkyl, -CONH ₂ , -alkylOH, -CN, -NO ₂ , halogen, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, - OH, -Oalkyl, -CO ₂ H, -CONH ₂ , -CO ₂ alkyl, halogen, -CF ₃ and -OCF ₃ , more especially hydrogen, -OH, -Oalkyl, -CO ₂ H and -CO ₂ alkyl;

R ₇ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl and aryl, especially hydrogen and alkyl, more especially hydrogen and methyl; n is an integer from 1 to 4, especially 1 to 3, more especially 1 or 2, most especially 2;

One of R ₈ and R ₁₂ is hydrogen and the other is selected from hydrogen, alkyl, cycloalkyl, cycloalkenyl, aryl, halogen, -OH, -Oalkyl, -Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , - N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, halogen, -OH, -Oalkyl, - NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen and -OH, most especially hydrogen;

All of V, W, X, Y and Z are -C= an one of R9 and R ₁₁ is an electron pair donor selected from -OH, -Oalkyl, -Oalkenyl, -Oalkynyl, -Ocycloalkyl, -Ocycloalkenyl, -Oaryl,

-Oheterocyclyl, -Oheteroaryl, -OCF ₃ , -NH ₂ , -NH(alkyl), -NH(alkenyl), -NH(alkynyl), -NH(cycloalkyl), -NH(cycloalkenyl), -NH(aryl), -NH(heterocyclyl), -NH(heteroaryl), -N(alkyl) ₂ , -N(cycloalkyl) ₂ , -N(cycloalkenyl) ₂ -N(aryl) ₂ , -N(alkyl)(aryl),

-N(alkyl)(cycloalkyl), -N(alkyl)(cycloalkenyl), -N(aryl)(cycloalkyl) and

N(aryl)(cycloalkenyl), especially -OH, -Oalkyl, -OCF ₃ , -NH ₂ , -NH(alkyl) and -N(alkyl) ₂ , more especially -OH, -OCH ₃ , -OCF ₃ , -NH ₂ , -NH(CH ₃ ) and -N(CH ₃ ) ₂ , most especially -OH, -OCH ₃ and -NH ₂ , and the other is selected from hydrogen, alkyl, cycloalkyl, aryl, halogen, -OH, -Oalkyl, -Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , -N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, halogen, -OH, -Oalkyl, -NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen and -OH, most especially hydrogen; or one of V, W, X, Y and Z is -0-, -S-, -NH- or -N= and the others are independently selected from -C= and -N=, an R9 and R11 are independently selected from hydrogen, alkyl, cycloalkyl, aryl, halogen, -OH, -Oalkyl, -Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , -N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, halogen, -OH, -Oalkyl, -NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen and -OH, most especially hydrogen and R ₁₀ is selected from hydrogen, alkyl, cycloalkyl, cycloalkenyl, aryl, halogen, -OH, -Oalkyl, -Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , -N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF3, especially hydrogen, alkyl, halogen, -OH, -Oalkyl, -NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen, -OH, -OCH ₃ and -OCF ₃ .

[0054] In some embodiments, in step (i) the optionally substituted alkylamine is selected from 3-amino-l-phenyl-propanol, 3-hydroxy-4-methoxy-phenylethylamine, 6- hydroxydopamine, 3-methoxytyramine, phenylethylamine, 3 -phenyl- 1 -propylamine, tryptamine, dopamine, serotonin, octapamine, tyramine, norepinephrine, 2-(l- naphthyl)ethylamine and 3-(l-naphthyl)propylamine and the optionally substituted heteroaryl or aryl alkylamine of step (ii) is selected from D/L-DOPA, 3-hydroxy-4- methoxyphenylethylamine, N-methylhomoveratylamine, 3-methoxytyramine, epinephrine, phenylethylamine, tryptamine, dopamine, serotonin, octapamine, tyramine and

norepinephrine. In further embodiments thereof, the carbonyl-containing co-substrate of step (ii) is a ketovacid,such as pyruvate, acetone, oxaloacetic acid, a-ketoglutarate, glyoxylic acid, acetoacetic acid and levulinic acid. [0055] In particular embodiments, the compound of formula (III) is selected from D/L- DOPA, 3-hydroxy-4-methoxyphenylethylamine, N-methylhomoveratylamine, 3- methoxytyramine, epinephrine, phenylethylamine, tryptamine, dopamine, serotonin, octapamine, tyramine and norepinephrine.

[0056] The steps i) and ii) of the disclosure may be carried out in any reaction medium and at any pH that solubilizes the biocatalysts, substrates and co-substrate used in the reactions. In some embodiments, the reaction medium is an aqueous reaction medium, for example, water or buffer, especially buffer. The buffer may have a pH that optimises one or both of the biocatalytic reactions or is suitable for both biocatalysts to function.

Suitable pH is in the range of 6 to 12, especially 7 to 10, more especially 7 to 9, for example, about 8. Suitable buffers to achieve these pH levels are known to those in the art and include, but are not limited to, Tris buffer, phosphate buffer and bicarbonate buffer.

[0057] In some embodiments, the reaction medium further comprises an antioxidant. The antioxidant may be derived from the compositions of the optionally substituted alkyl amine and/or aryl or heteroaryl alkylamine used to prepare the reaction mixture. The presence of an antioxidant in the compositions of optionally substituted alkylamine or aryl or heteroaryl alkylamine may assist in maintaining stability. The antioxidant may be any antioxidant that does not inhibit the activity of the biocatalysts or react with the substrates or co-substrate. Suitable antioxidants include sodium metabisulfite and ascorbic acid. The antioxidant may be present in the reaction mixture of biocatalyst reactions in an amount of 0.1 to 1.5% w/v.

[0058] The reaction mixture in which the optionally substituted alkyl amine, aryl or heteroaryl alkylamine and biocatalyst are contacted may be mixed by any suitable means, such as shaking or stirring for a time suitable for completion of the reaction to occur. The reaction may be followed by periodic sampling and analysis by chromatographic methods such as thin layer chromatography (TLC), Liquid Chromatography-Mass Spectrometry (LC-MS) or High Performance Liquid Chromatography (HPLC) to determine the extent of consumption of the substrates for each biocatalyst and appearance of product.

[0059] In some embodiments, the disclosure is most conveniently carried out at ambient temperature, especially between 15 °C and 30 °C. [0060] The compounds produced by the methods of the present disclosure include at least one new asymmetric centre at the position adjacent to the newly formed ring junction carbon atom, for example, the 1 position of an isoquinoline group. The new asymmetric centre produced may be in the R and/or S configuration depending on the Pictet- Spenglerase biocatalyst used. In some embodiments, the new asymmetric centre produced is in the S configuration. In other embodiments, the new asymmetric centre produced is in the R configuration. In yet other embodiments, the new asymmetric centre has a proportion of both R and S configuration, for example, a racemic mixture.

[0061] In some embodiments, it may be practical to recover one or both of the biocatalysts after performing the methods of the disclosure so that they can be reused. Recovery of the biocatalyst may be achieved as known in the art, for example, by immobilising the biocatalyst to a solid inert support such as glass or alginate beads or another inert matrix. One or both of the biocatalysts may then be recovered from the product mixture, for example, by filtration, and recycled into further biotransformation reactions. Advantageously, removal of the biocatalysts from the reaction mixture also simplifies recovery of the desired product from the reaction mixture.

[0062] If necessary, the compounds produced by the methods of the disclosure may be purified to provide a substantially pure product. Purification may be achieved by methods known in the art for separating organic molecules such as l-substituted-2N-containing fused heteropolycyclic compounds and the amino acid byproduct, if produced in the transamination reaction. For example, the products may be purified by acidification and solvent extraction, crystallization and/or chromatographic separation as known in the art.

Compounds Produced by the Methods

[0063] In some embodiments, the compounds produced by the methods of the disclosure are compounds of formula (VII):

wherein Ar is an optionally substituted aryl group further substituted with an electron pair donor in a position β to the ring junction carbon atom labeled 4, or an optionally substituted heteroaryl group;

Ri is selected from -CH ₃ and an optionally substituted aryl or heteroaryl group;

Each R ₂ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ;

Each R ₃ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl and heteroaryl;

Each R ₄ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl and -OR ₃ ;

Each R5 is independently selected from hydrogen and halogen;

Each R ₆ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ ,

-OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and

-OC(R ₅ ) ₃ ;

R ₇ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, -OH, -Oalkyl, -Oalkenyl and -Oalkynyl;

n is an integer from 1 to 6, such as 1, 2, 3, 4, 5, or 6; and

p is an integer from 0 to 12, such as 1,2,3,4, 5, 6, 7, 8, 9, 10, 11 or 12. [0064] In some embodiments, the Ar group is an optionally substituted heteroaryl group in which at least one substituent is an electron pair donor.

[0065] In some embodiments, the compound of formula (VII) is a compound of formula (VIII):

wherein Ri, R ₂ , R ₃ , R ₄ , R5, R ₆ , R7, n and p are as defined in formula (VII); one of W, X, Y and Z is selected from -0-, -S-, -NH- and -N= and the others are independently selected from -C= and -N= or each of W, X, Y and Z are independently selected from -C= and -N=; q is 0 or 1 ;

Each R ₈ , R9, R ₁₀ and R ₁₁ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or when all of W, X, Y and Z are -C=, R9 is an electron pair donor and each of R ₈ , R ₁₀ and R ₁₁ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or R ₁₀ and R ₁₁ or R ₁₀ and R9 or R ₈ and R9 together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; with the proviso that (i) When any of W, X, Y and Z are -0-, -S-, -NH- or -N=, their substituent R ₁₁ , R ₁₀ , R9 and R ₈ respectively is absent.

[0066] In particular embodiments, the compound of formula (VIII) is an aryl compound of formula (IX):

wherein Ri, R ₂ , R ₃ , R ₄ , R5, R ₆ , R7, n and p are defined as for formula (VIII);

R ₈ , R ₁₀ and R ₁₁ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and -OC(R ₅ ) ; and

R9 is an electron pair donor.

[0067] In particular embodiments of compounds of formula (VII), (VIII) and (IX), one or more of the following applies:

Ri is selected from -CH ₃ , phenyl, naphthyl, tetrahydronaphthyl, pyridinyl, pyrimidinyl, pyrazinyl, pyrrolyl, pyrazolyl, imidazolyl, indolyl, benzofuranyl, benzothiophenyl, indolinyl, isoindolyl, benzimidazolyl, quinalinyl, isoquinolinyl, quinazolinyl, quinoxalinyl and naphthyridinyl, especially phenyl, naphthyl, pyridinyl, quinolinyl and isoquinolinyl; each R ₂ is independently selected from hydrogen, alkyl, halogen, -OH, -Oalkyl, -CO ₂ H, -CO ₂ alkyl, -CONH ₂ , -alkyl-OH, -NH ₂ , -CF ₃ and -OCF ₃ , especially hydrogen, hydroxyl or -CO ₂ H, more especially hydrogen or hydroxyl; p is an integer from 1 to 10, especially 1 to 8 or 1 to 6, more especially 1 to 4, most especially 1 or 2; Each R ₆ is independently selected from hydrogen, alkyl, -OH, -Oalkyl, -CO ₂ H, -CO ₂ alkyl, -CONH ₂ ,-alkylOH, -CN, -NO ₂ , halogen, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, -OH, -Oalkyl, -CO ₂ H, -CO ₂ alkyl, -CONH ₂ , halogen, -CF ₃ and -OCF ₃ , more especially hydrogen, -OH, -Oalkyl, -CO ₂ H and -CO ₂ alkyl;

R ₇ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl and aryl, especially hydrogen and alkyl, more especially hydrogen and methyl; n is an integer from 1 to 4, especially 1 to 3, more especially 1 or 2, most especially 2;

One of W, X, Y and Z is -0-, -S-, -NH- or -N= and the others are independently selected from -C= and -N= or W, X, Y and Z are all -C=; q is 0 or 1 ;

R ₈ is selected from hydrogen, alkyl, cycloalkyl, cycloalkenyl, aryl, halogen, -OH, -Oalkyl, -Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , -N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, halogen, -OH, -Oalkyl, -NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen and -OH, most especially hydrogen;

R9 is selected from hydrogen, alkyl, cycloalkyl, aryl, halogen, -OH, -Oalkyl, -Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , -N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, halogen, -OH, -Oalkyl, -NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen and -OH, most especially hydrogen; or when each of W, X, Y and Z are all -C=, R9 is an electron pair donor selected from -OH, Oalkyl, -Oalkenyl, -Oalkynyl, -Ocycloalkyl, -Ocycloalkenyl, -Oaryl, -Oheterocyclyl, -Oheteroaryl, -OCF ₃ , -NH ₂ , -NH(alkyl), -NH(alkenyl), -NH(alkynyl), -NH(cycloalkyl), -NH(cycloalkenyl), -NH(aryl), -NH(heterocyclyl), -NH(heteroaryl), -N(alkyl) ₂ ,

-N(cycloalkyl) ₂ , -N(cycloalkenyl) ₂ -N(aryl) ₂ , -N(alkyl)(aryl), -N(alkyl)(cycloalkyl), -N(alkyl)(cycloalkenyl), -N(aryl)(cycloalkyl) and N(aryl)(cycloalkenyl), especially -OH, -Oalkyl, -OCF ₃ , -NH ₂ , -NH(alkyl) and -N(alkyl) ₂ , more especially -OH, -OCH ₃ , -OCF ₃ , -NH ₂ , -NH(CH ₃ ) and -N(CH ₃ ) ₂ , most especially -OH, -OCH ₃ and -NH ₂ ; R ₁₀ is selected from hydrogen, alkyl, cycloalkyl, cycloalkenyl, aryl, halogen, -OH, -Oalkyl, -Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , -N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, halogen, -OH, -Oalkyl, -NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen, -OH, -OCH ₃ and -OCF ₃ ; and

Rii is selected from hydrogen, alkyl, cycloalkyl, aryl, halogen, -OH, -Oalkyl,

-Ocycloalkyl, -Oaryl, -NH ₂ , -NHCH ₃ , -N(CH ₃ ) ₂ , -alkylOH, -CF ₃ and -OCF ₃ , especially hydrogen, alkyl, halogen, -OH, -Oalkyl, -NH ₂ , -CF ₃ and -OCF ₃ , more especially hydrogen and -OH, most especially hydrogen.

[0068] In yet another aspect of the disclosure there is provided a compound of formula

(X):

wherein Ar is an optionally substituted aryl or heteroaryl group; r is 0 or an integer from 1 to 4, such as 1, 2, 3, or 4;

Each of R ₂₀ , R ₂₂ and R ₂₃ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₂₈ , -CO ₂ R ₂₈ , -C(O)R ₂₈ , -alkyl-OH, -C(O)N(R ₂₈ ) ₂ , -N(R ₂₉ ) ₂ , -C(R ₃₀ ) ₃ and -OC(R ₃₀ ) ₃ ;

R ₂₁ is an electron pair donor;

Each of R ₂₄ and R ₂ 5 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₂₈ , -CO ₂ R ₂₈ , -C(O)R ₂₈ , -alkyl-OH, -C(O)N(R ₂₈ ) ₂ , -N(R ₂₉ ) ₂ , -C(R ₃₀ ) ₃ and -OC(R ₃₀ ) ₃ ;

R ₂ 6 is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, -OH, -Oalkyl, Oalkenyl and -Oalkynyl; Each R27 is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR28, -CO2R28,

-C(O)R ₂₈ , -alkyl-OH, -C(O)N(R ₂₈ ) ₂ , -N(R ₂₉ ) ₂ , -C(R ₃₀ ) ₃ and -OC(R ₃₀ ) ;

Each R ₂₈ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl and heteroaryl;

Each R29 is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl and -OR2 ₈ ;

Each R ₃ o is independently selected from hydrogen and halogen; with the proviso that: when R20, R23, R24, R25, R26 and R27 are hydrogen, R21 and R22 are -OH and r is 1, Ar is not unsubstituted phenyl, 4-hydroxyphenyl, 3,4-dihydroxyphenyl or 3-thiophenyl.

[0069] In some particular embodiments of formula (X), one or more of the following applies:

Ar is selected from phenyl, indole, furan,benzofuran, pyrrole, benzothiophenyl, pyridine, quinolone, isoquinoline and thiophenyl, each of which may be optionally substituted with one or more of -OH, -Oalkyl, -Oalkenyl, -Oalkynyl, halo, -CN-, -NO ₂ , -CF ₃ and -OCF ₃ ;

R20 is hydrogen, -OH or -Oalkyl, especially hydrogen;

R21 is an electron pair donor selected from -OH, -Oalkyl, -Oalkenyl, -Oalkynyl, - Ocycloalkyl, -Ocycloalkenyl, -Oaryl, -Oheterocyclyl, -Oheteroaryl, -OCF ₃ , -N¾,

-NH(alkyl), -NH(alkenyl), -NH(alkynyl), -NH(cycloalkyl), -NH(cycloalkenyl), -NH(aryl), -NH(heterocyclyl), -NH(heteroaryl), -N(alkyl) ₂ , -N(cycloalkyl) ₂ , -N(cycloalkenyl) ₂ , -N(aryl) ₂ , -N(alkyl)(aryl), -N(alkyl)(cycloalkyl), -N(alkyl)(cycloalkenyl),

-N(aryl)(cycloalkyl) and N(aryl)(cycloalkenyl), especially -OH, -Oalkyl, -OCF , -NH ₂ , -NH(alkyl) and -N(alkyl) ₂ , more especially -OH, -OCH ₃ , -OCF ₃ , -NH ₂ , -NH(CH ₃ ) and -N(CH ₃ ) ₂ , most especially -OH, -OCH ₃ and -NH ₂ ;

R22 is selected from hydrogen, -OH, -Oalkyl, -Oalkenyl, -Oalkynyl, halogen, -CN, -NO2, - NH2 -CF ₃ and -OCF ₃ , especially hydrogen, -OH, -Oalkyl, halogen, -OCF ₃ and -N¾, more especially hydrogen, hydroxyl, -OCH ₃ and -OCF ₃ ; R23 is hydrogen, -OH or -Oalkyl, especially hydrogen;

R ₂₄ is selected from hydrogen, -OH, -Oalkyl, -Oalkenyl, -Oalkynyl, halogen, -CN, -NO ₂ , - NH ₂ -CF ₃ and -OCF ₃ , especially hydrogen, -OH, -Oalkyl, halogen, -OCF ₃ and -NH ₂ , more especially hydrogen, hydroxyl, -OCH ₃ and -OCF ₃ ;

R ₂₅ is selected from hydrogen, -OH, -Oalkyl, -CO ₂ H, -CO ₂ alkyl, -C(O)alkyl, -C(O)H, - OCF ₃ , -NH ₂ and -CONH ₂ ; especially hydrogen, OH, -CO ₂ H, -CONH ₂ and -NH ₂ , more especially hydrogen, -OH and -CO ₂ H;

R ₂ 6 is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl and aryl, especially hydrogen and alkyl, more especially hydrogen and methyl;

R27 is selected from hydrogen, -OH, -Oalkyl, -Oalkenyl, -Oalkynyl, halogen, -CN, -NO ₂ , - NH ₂ -CF ₃ and -OCF ₃ , especially hydrogen, -OH, -Oalkyl, halogen, -OCF ₃ and -NH ₂ , more especially hydrogen, hydroxyl, -OCH ₃ and -OCF ₃ ; r is selected from 0, 1, 2 or 3, especially 1 or 2.

[0070] In some particular embodiments, the compound of formula (X) is selected from: l-[(indol-3-yl)methyl]-l,2,3,4-tetrahydroisoquinoline-6,7-di ol (compound A); l-[(5-hydroxyindol-3-yl)methyl]-l,2,3,4-tetrahydroisoquinoli ne-6,7-diol (compound B);

1 -[2-phenylethyl] -1,2,3 ,4-tetrahydroisoquinoline-6,7-diol (compound C) ; l-[(3-hydroxy-4-methoxyphenyl)methyl]-l,2,3,4-tetrahydroisoq uinoline-6,7-diol

(compound D); l-[(3,4-dihydroxyphenyl)methyl]-l,2,3,4-tetrahydroisoquinoli ne-6,7-diol (compound E); l-[2-hydroxy-2-phenylethyl]-l,2,3,4-tetrahydroisoquinoline-6 ,7-diol (compound F); l-[(4-hydroxy-3-methoxyphenyl)methyl]-l,2,3,4-tetrahydroisoq uinoline-6,7-diol

(compound G); l-[(3,4-dihydroxyphenyl)methyl]-7-hydroxy-6-methoxy-l,2,3,4- tetrahydroisoquinoline (compound H); l-[(4-hydroxyphenyl)methyl]-l,2,3,4-tetrahydroisoquinoline-4 ,6,7-triol (compound I); 1 - [(3 ,4-dihydroxyphenyl)methyl] - 1 ,2,3 ,4-tetrahydroisoquinoline-4,6,7-triol (compound J) ; l-[(5-hydroxyindol-3-yl)methyl]-l,2,3,4-tetrahydroisoquinoli ne-4,6,7-triol (compound K); l-[(indol-3-yl)methyl]-l,2,3,4-tetrahydroisoquinoline-4,6,7- triol (compound L); l-[2-phenylethyl]-l,2,3,4-tetrahydroisoquinoline-4,6,7-triol (compound M); l-[phenylmethyl]-l,2,3,4-tetrahydroisoquinoline-4,6,7-triol (compound N); l-[(3-hydroxy-4-methoxyphenyl)methyl]-l,2,3,4-tetrahydroisoq uinoline-4,6,7-triol (compound O); l-[2-hydroxy-2-phenylethyl]-l,2,3,4-tetrahydroisoquinoline-4 ,6,7-triol (compound P); l-[(4-hydroxy-3-methoxyphenyl)methyl]-l,2,3,4-tetrahydroisoq uinoline-4,6,7-triol (compound Q);

[l-(4-hydroxyphenylmethyl)-6,7-dihydroxy-l,2,3,4-tetrahyd roisoquinolin-3-yl] carboxylic acid (compound R);

[l-(4-hydroxy-3-methoxyphenylmethyl)-6,7-dihydroxy-l,2,3, 4-tetrahydroisoquinolin-3-yl] carboxylic acid (compound S);

[l-(3-hydroxy-4-methoxyphenylmethyl)-6,7-dihydroxy-l,2,3, 4-tetrahydroisoquinolin-3-yl] carboxylic acid (compound T);

[l-(indol-3-ylmethyl)-6,7-dihydroxy-l,2,3,4-tetrahydroiso quinolin-3-yl] carboxylic acid (compound U);

[l-(5-hydroxyindol-3-ylmethyl)-6,7-dihydroxy-l,2,3,4-tetr ahydroisoquinolin-3-yl] carboxylic acid (compound V);

[l-(phenylmethyl)-6,7-dihydroxy-l,2,3,4-tetrahydroisoquin olin-3-yl] carboxylic acid (compound W);

[l-(2-phenylethyl)-6,7-dihydroxy-l,2,3,4-tetrahydroisoqui nolin-3-yl] carboxylic acid (compound X);

[l-(2-hydroxy-2-phenylethyl)-6,7-dihydroxy-l,2,3,4-tetrah ydroisoquinolin-3-yl] carboxylic acid (compound Y); l-[(5-hydroxyindol-3-yl)methyl]-6-hydroxy-7-methoxy-l,2,3,4- tetrahydroisoquinoline (compound Z); l-[(indol-3-yl)methyl]-6-hydroxy-7-methoxy-l,2,3,4-tetrahydr oisoquinoline (compound

AA); l-[(3-hydroxy-4-methoxyphenyl)methyl]-6-hydroxy-7-methoxy- 1,2,3,4- tetrahydroisoquinoline (compound AB); l-[(indol-3-yl)methyl]-2N-methyl-l,2,3,4-tetrahydroisoquinol ine-4,6,7-triol (compound

AC); and l-[2-phenylethyl]-2N-methyl-l,2,3,4-tetrahydroisoquinoline-4 ,6,7-triol (compound AD). Uses of the Compounds made by the Methods

[0071] Pictet-Spenglerase biocatalyst mediated reactions are important in the synthesis of many alkaloid based scaffolds (Stockigt et al., Angew. Chem. Int. Ed., 2011, 50, 8538- 8564). The methods of the disclosure can be used to prepare useful compounds efficiently and under mild conditions and on an industrially useful scale. The compounds produced may be an intermediate product that can be sold and/or used as an intermediate in the production of further more complex industrially useful compounds such as

pharmaceuticals. In other cases, the compounds produced may be biologically active and therefore pharmaceutically useful in their own right.

[0072] In another aspect of the disclosure there is provided a use of a compound of formula (VII)

wherein Ar is an optionally substituted aryl group further substituted with an electron pair donor in a position β to the ring junction carbon atom labeled 4, or an optionally substituted heteroaryl group;

Ri is selected from -CH ₃ and an optionally substituted aryl or heteroaryl group;

Each R ₂ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and -OC(R ₅ ) ;

R ₃ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl and heteroaryl;

Each R ₄ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl and -OR ₃ ;

Each R5 is independently selected from hydrogen and halogen;

Each R ₆ is independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and -OC(R ₅ ) ;

R ₇ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, -OH, -Oalkyl, -Oalkenyl and -Oalkynyl;

n is an integer from 1 to 6, such as 1, 2, 3, 4, 5, or 6; and p is an integer from 0 to 12, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; as an intermediate in the synthesis of a biologically active compound.

[0073] In some embodiments, Ar is an optionally substituted heteroaryl group in which at least one optional substituent is an electron pair donor.

[0074] In some embodiments, the compound of formula (VII) is a compound of formula (VIII):

wherein R ₁ , R ₂ , R ₃ , R ₄ , R5, R ₆ , R7, n and p are as defined in formula (VII); one of W, X, Y and Z is selected from -0-, -S-, -NH- and -N= and the others are independently selected from -C= and -N= or each of W, X, Y and Z are independently selected from -C= and -N=; q is 0 or 1 ;

Each R ₈ , R9, R ₁₀ and R ₁₁ is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or when all of W, X, Y and Z are -C=, R9 is an electron pair donor and each of R ₈ , R ₁₀ and R ₁₁ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) ₃ and -OC(R ₅ ) ₃ ; or R ₁₀ and R ₁₁ or R ₁₀ and R9 or R ₈ and R9 together with the atoms to which they are attached form an optionally substituted carbocyclic or heterocyclic saturated, unsaturated or aromatic ring; with the proviso that

When any of W, X, Y and Z are -0-, -S-, -NH- or -N=, their substituent R ₁₁ , R ₁₀ , R9 and R ₈ respectively is absent.

[0075] In some particular embodiments, the compound of formula (VIII) is a compound of formula (IX):

wherein Ri, R ₂ , R ₃ , R ₄ , R5, R ₆ , R7, n and p are defined as for formula (VIII);

R ₈ , R ₁₀ and R ₁₁ are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, halogen, -CN, -NO ₂ , -OR ₃ , -SR ₃ , -CO ₂ R ₃ , -C(O)R ₃ , -alkyl-OH, -C(O)N(R ₃ ) ₂ , -N(R ₄ ) ₂ , -C(R ₅ ) and -OC(R ₅ ) ; and

R9 is an electron pair donor.

[0076] The biologically active compound that is derived from the compounds produced by the methods of the disclosure may be alkaloids such as isoquinoline derived alkaloids. For example, ?-reticuline is a synthetic precursor in the preparation of morphine and other related alkaloids and may be prepared by the method of the present disclosure using 3-hydroxy-4-methoxyphenylethylamine and N-methyl-3-hydroxy-4-methoxyphenyl ethylamine in a single one pot reaction in aqueous medium and at ambient temperature.

[0077] The compounds produced by the methods of the disclosure may also have useful biological activity in humans or animals without further modification.

[0078] In a further aspect of the disclosure there is provided a l-substituted-2N- containing fused heteropolycyclic compound, especially an optionally substituted alkyl- 1,2,3,4-tetrahydroisoquinoline compound prepared by the method of the disclosure.

[0079] Norcoclaurine, also known as Higenamine or l-[(4-hydroxyphenyl)methyl]- l,2,3,4-tetrahydroisoquinoline-6,7-diol, has been found to have β-2- adrenergic receptor agonist activity and has been investigated for its vasodilatory, cardiotonic and

bronchodilatory activity in humans and animals. The method of the present disclosure is able to produce Norcoclaurine from commercially available amine compounds dopamine and tyramine in a single one pot reaction.

[0080] Some embodiments of the disclosure will now be further described by the following Examples, which are included for the purpose of illustration only and are not to be construed as limiting the generality hereinbefore described.

EXAMPLES

Example 1: General Method of Synthesis of 1,2,3,4-tetrahydroisoquinolines

[0081] 100 μΐ _^ of Substrate 1 (arylalkyamine compound) (50 mM stock solution in 1% metabisulfite solution), 100 μΐ _^ of Substrate 2 (substituted aryl alkylamine compound) (50 mM stock solution in 1% metabisulfite solution) and 100 μΐ _^ of pyruvate (50 mM aqueous solution was combined in Tris buffer (100 mM, pH 8) with a Pictet-Spenglerase (NCS from Thalictrum flavum (~ 1 μg) and a transaminase biocatalyst (~ 1 μg) (SEQ ID NO: 1 or SEQ ID NO: 4 from Pseudomonas spp.).

[0082] If the transaminase biocatalyst accepts a co-substrate other than pyruvate, the pyruvate is interchangeable with other co-substrates.

[0083] The mixture was left shaking overnight at room temperature and then assayed by liquid chromatography-mass spectrometry (LC-MS). LC-MS was carried out using an Astec Chirobiotic™ T chiral CPLC column (5 μιη by 25 cm x 4.6 mm) using a mobile phase of methanol : water : formic acid (70 : 30 : 0.02% w/w) at 1 niL/minute for 20 minutes.

[0084] Products were identified by TOF mass spectrometry on an Agilent 1100 system.

[0085] The results for varying substrates are shown below. The reactions shown below indicate that the transaminase reaction used a biocatalyst with SEQ ID NO:4. However, the same reactions were performed with a biocatalyst of SEQ ID NO:2 and the same results were obtained.



Substrate 1: 3-Hydroxy-4-methoxyphenylethylamine or Dopamine Substrate 2: 3-Hydroxy-4-methoxyphenylethylamine or Dopamine TOF-MS: M ^-1 315

Only one compound was prepared but it was not possible to distinguish between the two possible products by mass spectrometry.

some embodiments the following conditions set out in the table below