Polysubstituted psilocybin derivatives and methods of use thereof

Abstract

The present disclosure relates to novel poly-substituted psilocybin derivative compounds and pharmaceutical or recreational drug formulations containing such compounds, which are produced by reacting a reactive psilocybin derivative with a substituent-containing compound.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 149,001, filed February 12, 2021, and U.S. Provisional Application No. 63 / 247,881, filed September 24, 2021, which are incorporated herein by reference in their entireties. [Technical Field]

[0002] The compositions and methods disclosed herein relate to the compound known as psilocybin. Additionally, the compositions and methods disclosed herein relate specifically to psilocybin derivatives having multiple substituents. [Background technology]

[0003] The following sections are provided as background to the present disclosure, but are not to be construed as an admission that anything discussed herein is part of the prior art or the knowledge of those skilled in the art.

[0004] Biochemical pathways in living cells can be classified as part of primary metabolism or part of secondary metabolism. Pathways that are part of a cell's primary metabolism are those involved in catabolism for energy production or anabolism for the production of cellular infrastructure. Secondary metabolism, on the other hand, is produced by cells without any apparent anabolic or catabolic function. Secondary metabolism has long been recognized as beneficial in many ways, including the production of therapeutic compounds.

[0005] Psilocybin is a secondary metabolite naturally produced by certain mushrooms, which can be classified as part of the Basidiomycota phylum within the fungal kingdom. Mushroom species capable of producing psilocybin include species of the genus Psilocybe, such as Psilocybe azurescens, Psilocybe semilansata, Psilocybe cervica, Psilocybe mexicana, and Psilocybe cyanescens. Interest in psilocybin is well established in the industry. Thus, for example, psilocybin is a psychotropic compound and therefore also used as a recreational drug. Furthermore, psilocybin has been used as a research tool in behavioral and neuroimaging studies of neurological disorders and has been evaluated for its clinical potential in the treatment of mental health conditions (Non-Patent Document 1), including the treatment of anxiety in terminal cancer patients (Non-Patent Document 2) and the alleviation of symptoms of treatment-resistant depression (Non-Patent Document 3). [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Daniel, J. et al., Mental Health Clin / , 2017;7(1): 24-28 [Non-patent document 2] Grob, C. et al., Arch. Gen. Psychiatry, 2011, 68(1) 71-78 [Non-patent document 3] Cathart-Harris, RL et al., Lancet Psychiatry, 2016, 3: 619-627 Summary of the Invention [Problem to be solved by the invention]

[0007] Although psilocybin has low toxicity, recreational users often experience side effects including panic attacks, paranoia, and psychotic states, sometimes collectively or individually referred to as "bad trips."

[0008] Thus, there is a need in the art for improved psilocybin compounds. [Means for solving the problem]

[0009] The following sections are intended to introduce the reader to further details, but are not intended to define or limit the subject matter claimed in this disclosure.

[0010] In one embodiment, the present disclosure relates to psilocybin and its derivative compounds.

[0011] In another aspect, the disclosure relates to psilocybin derivative compounds and methods of making and using such compounds.

[0012] In another aspect, the disclosure relates to poly-substituted psilocybin derivative compounds.

[0013] Thus, in one aspect, the present disclosure provides, in at least one embodiment, in accordance with the teachings herein, a compound of the following formula: [ka] (wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3cis a hydrogen atom or a carboxyl group), or a salt thereof.

[0014] In at least one embodiment of one aspect, at least three of R2, R4, R5, R6, or R7 may be a substituent selected from at least two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0015] In at least one embodiment of one aspect, at least three of R2, R4, R5, R6, or R7 may be a substituent selected from at least three of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0016] In at least one embodiment of one aspect, when R4 is not substituted with a substituent, R4 may be a hydrogen atom.

[0017] In at least one embodiment of one aspect, R4 and R5 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and R 2、 R6 and R7 may be hydrogen atoms.

[0018] In at least one embodiment of one aspect, R4 and R5 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and at least one of R4 and R5 may be a prenyl group or a hydrogen atom, and R 2、 R6 and R7 may be hydrogen atoms.

[0019] In at least one embodiment of one aspect, R4 and R6 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and R 2、 R4 and R7 may each be a hydrogen atom.

[0020] In at least one embodiment of one aspect, R4 and R6 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and at least one of R4 and R6 may be a prenyl group or a hydrogen atom, and R 2、 R4 and R7 may each be a hydrogen atom.

[0021] In at least one embodiment of one aspect, R4 and R7 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and R 2、 R5 and R6 may be hydrogen atoms.

[0022] In at least one embodiment of one aspect, R4 and R7 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and at least one of R4 and R7 may be a prenyl group or a hydrogen atom, and R 2、 R5 and R6 may be hydrogen atoms.

[0023] In at least one embodiment of one aspect, R5 and R6 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and R 2、 R4 and R7 may each be a hydrogen atom.

[0024] In at least one embodiment of one aspect, R5 and R7 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and at least one of R5 and R6 may be a prenyl group or a hydrogen atom, and R 2、 R4 and R6 may be hydrogen atoms.

[0025] In at least one embodiment of one aspect, R5 and R7 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and R 2、 R4 and R6 may be hydrogen atoms.

[0026] In at least one embodiment of one aspect, R5 and R7 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and at least one of R5 and R7 may be a prenyl group or a hydrogen atom, and R 2、 R4 and R6 may be hydrogen atoms.

[0027] In at least one embodiment of one aspect, R6 and R7 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and R 2、 R4 and R5 may be hydrogen atoms.

[0028] In at least one embodiment of one aspect, R6 and R7 may be selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and at least one of R6 and R7 may be a prenyl group or a hydrogen atom, and R 2、 R4 and R5 may be hydrogen atoms.

[0029] In at least one embodiment of one aspect, R2 may be hydrogen, in which case only two of R4, R5, R6, or R7 are substituted, and the first substituent is selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and the first and second substituents are different groups.

[0030] In at least one embodiment of one aspect, R2 may be hydrogen and only two of R4, R5, R6, or R7 are substituted, in which case the first substituent is a hydrogen atom and the second substituent is selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, and (viii) a nitrile group.

[0031] In at least one embodiment of one aspect, R2 may be hydrogen and only two of R4, R5, R6, or R7 are substituted, in which case the first substituent is a prenyl group and the second substituent is selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, and (viii) a nitrile group.

[0032] In at least one embodiment of one aspect, R2 may be hydrogen and only two of R4, R5, R6, or R7 are substituted, in which case the first substituent is a nitrile group and the second substituent is selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, and (viii) a prenyl group.

[0033] In at least one embodiment of one aspect, R2 may be hydrogen and only two of R4, R5, R6, or R7 are substituted, in which case the first substituent is selected from (i) a halogen atom, (ii) a prenyl group, and (iii) a nitrile group, and the second substituent is selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, or (vi) an aldehyde or ketone group.

[0034] In at least one embodiment of one aspect, R2 may be hydrogen and only two of R4, R5, R6, or R7 are substituted, in which case the first substituent is a hydrogen atom and the second substituent is selected from (i) an amino group or an N-substituted amino group, (ii) a nitrile group, (iii) a nitro group, (iv) a hydroxy group, and (v) a prenyl group.

[0035] In at least one embodiment of one aspect, R2 may be hydrogen and only two of R4, R5, R6, or R7 may be substituted, in which case the first substituent is a prenyl group and the second substituent is selected from (i) a carboxyl group or a carboxylic acid derivative, (ii) a halogen, and (iii) a hydroxy group, in which case R2 may be hydrogen and two of R4, R5, R6, or R7 may be substituted.

[0036] In at least one embodiment of one aspect, R2 may be hydrogen, and only two of R4, R5, R6, or R7 are substituted, in which case the first substituent is a halogen atom and the second substituent is a prenyl group.

[0037] In at least one embodiment of one aspect, R2 can be hydrogen and only two of R4, R5, R6, or R7 are substituted, in which case the first substituent is a nitrile group and the second substituent is an amino group or an N-substituted amino group.

[0038] In at least one embodiment of one aspect, R5 may be a carboxyl group or an acetyl group, and R7 may be an amino group, a nitrile group, a hydroxy group, or a halogen.

[0039] In at least one embodiment of one aspect, R5 may be an acetamidyl group and R7 may be an aldehyde group, a carboxyl group, or a carboxy ester.

[0040] In at least one embodiment of one aspect, R5 may be an acetamidyl group, R6 may be an amino group, a nitro group, or a halogen, and R7 may be an aldehyde group, a carboxyl group, or a carboxy ester.

[0041] In at least one embodiment of one aspect, R5 may be a carboxymethyl group, and R7 may be a nitro group, an amino group, or a halogen.

[0042] In at least one embodiment of one aspect, R4 may be a glycosyloxy group, R5 may be a carboxymethyl group or an amido group, and R7 may be a nitro group, an amino group, or a halogen.

[0043] In at least one embodiment of one aspect, compound (I) is represented by formula (IX), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI), (XXII), (XXIII), (XXIV), (XXV), (XXVI), (XXVII), (XXVIII), (XXIX), (XXX), (XXXI) , (XXXII), (XXXIII), (XXXIV), (XXXV), (XXXVI), (XXXVII), (XXXVIII), (XXXIX), (XL), (XLI), (XLII), (XLII I), (XLIV), (XLV), (XLVI), (XLVII), (XLVIII), (XLIX), (L), (LI), (LII), (LIII), (LIV), (LV), or (LXXVI): [ka] [ka] [ka] The compound may be selected from the following compounds:

[0044] In at least one embodiment of one aspect, compound (I) is any one of the compounds shown in Figures 13A, 13B, and 13C and designated herein as 13A-3, 13A-4, 13A-5, 13A-6, 13A-7, 13A-8, 13A-9, 13A-10, 13B-3, 13B-4, 13B-5, 13B-6, 13B-7, 13B-8, 13B-8, 13C-6, 13C-7, 13C-8, 13C-9, 13C-10, or 13C-11.

[0045] In another aspect, the present disclosure relates to pharmaceutical and recreational drug formulations containing psilocybin derivative compounds. Accordingly, in at least one embodiment of one aspect, the present disclosure provides a compound having the following formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group) or a salt thereof, together with a pharmaceutically acceptable excipient, diluent, or carrier.

[0046] In another aspect, the present disclosure relates to a method for treating a psychiatric disorder. Accordingly, in one embodiment, the present disclosure relates to a method for treating a psychiatric disorder, namely, a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group, and the pharmaceutical formulation is administered in an amount effective to treat the psychiatric disorder in the subject) or a salt thereof to a subject in need thereof.

[0047] In at least one embodiment of one aspect, the disorder is 5-HT 2A Receptor-mediated disorders or 5-HT 1A It may be a receptor-mediated disorder.

[0048] In at least one embodiment of one aspect, a dose of about 0.001 mg to about 5,000 mg is administered.

[0049] In another aspect, the present disclosure provides, in at least one embodiment, 5-HT 2A Receptor-mediated disorders or 5-HT 1A A method for modulating a receptor-mediated disorder, comprising administering to a subject a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group) or its salts with 5-HT 2A receptors or 5-HT 1A The method comprises contacting a receptor.

[0050] In at least one embodiment of one aspect, the reaction conditions may be in vitro reaction conditions.

[0051] In at least one embodiment of one aspect, the reaction conditions may be in vivo reaction conditions.

[0052] In another aspect, the present disclosure relates to a method of making a poly-substituted psilocybin derivative compound. Accordingly, in at least one embodiment of one aspect, the present disclosure relates to a method of making a poly-substituted psilocybin derivative compound having the following formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group) or a salt thereof, the method comprising: [ka] wherein one of R2, R4, R5, R6, or R7 is a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom or an alcohol group; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, a hydroxy group, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3cis a hydrogen atom or a carboxyl group) with a substituent-containing compound, wherein the substituent of the substituent-containing compound is selected from (i) halogen-containing compounds, (ii) hydroxy-containing compounds, (iii) nitro-containing compounds, (iv) glycosyloxy-containing compounds, (v) amino- or N-substituted amino-containing compounds, (vi) carboxyl- or carboxylic acid derivative-containing compounds, (vii) aldehyde- or ketone-containing compounds, (viii) prenyl-containing compounds, and (ix) nitrile-containing compounds, under reaction conditions sufficient to form a psilocybin substituent of formula (I) or a salt thereof.

[0053] In at least one embodiment of one aspect, the substituent of the reactive psilocybin derivative of formula (II) may be a nitro group, the substituent-containing compound may be a carboxylic acid derivative acetic anhydride (AcO), and the reactive psilocybin derivative and the substituent-containing compound may undergo a Friedel-Crafts acylation reaction to form a first psilocybin derivative of formula (I), wherein one of R, R, R, R, or R is a nitro group and at least one of R, R, R, R, R, or R is an acetyl group.

[0054] In at least one embodiment of one aspect, the first psilocybin derivative formed may be the compound shown in Figure 13A and representation 13A-3.

[0055] In at least one embodiment of one aspect, the formed first psilocybin derivative of formula (I) may be reacted to oxidize the acetyl group to form a second psilocybin derivative of formula (I), in which case one of R2, R4, R5, R6, or R7 may be a nitro group and at least one of R2, R4, R5, R6, or R7 may be a carboxyl group.

[0056] In at least one embodiment of one aspect, the second psilocybin derivative formed may be the compound shown in Figure 13A and representation 13A-4.

[0057] In at least one embodiment of one aspect, the formed second psilocybin derivative of formula (I) may react to reduce the nitro group to form an amino group and a third psilocybin derivative, wherein one of R2, R4, R5, R6, or R7 may be an amino group and at least one of R2, R4, R5, R6, or R7 may be a carboxyl group.

[0058] In at least one embodiment of one aspect, the third psilocybin derivative formed may be the compound shown in Figure 13A and designation 13A-5.

[0059] In at least one embodiment of one aspect, the formed tertiary psilocybin derivative of formula (I) may be reacted with a nitrite salt to convert the amino group in the diazonium salt to form an intermediate psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be a diazonium group and at least one of R2, R4, R5, R6, or R7 may be a carboxyl group.

[0060] In at least one embodiment of one aspect, the intermediate psilocybin derivative formed may be the compound shown in Figure 13A and representation 13A-6.

[0061] In at least one embodiment of one aspect, the formed intermediate psilocybin derivative of formula (I) may be reacted with a nitrile-containing compound to convert the diazonium group to form a fourth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be a nitrile group and at least one of R2, R4, R5, R6, or R7 may be a carboxyl group.

[0062] In at least one embodiment of one aspect, the quaternary psilocybin derivative formed may be the compound shown in Figure 13A and designation 13A-7.

[0063] In at least one embodiment of one aspect, the formed intermediate psilocybin derivative of formula (I) may react with water to convert the diazonium group to form a fifth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be a hydroxy group and at least one of R2, R4, R5, R6, or R7 may be a carboxyl group.

[0064] In at least one embodiment of one aspect, the fifth psilocybin derivative formed may be the compound shown in Figure 13A and designation 13A-8.

[0065] In at least one embodiment of one aspect, the formed intermediate psilocybin derivative of formula (I) may be reacted with a halogen-containing compound to convert the diazonium group to form a sixth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be a halogen atom and at least one of R2, R4, R5, R6, or R7 may be a carboxyl group.

[0066] In at least one embodiment of one aspect, the sixth psilocybin derivative formed may be the compound shown in Figure 13A and designations 13A-9 or 13A-10.

[0067] In at least one embodiment of one aspect, the substituent of the reactive psilocybin derivative of formula (II) may be a methoxycarbonyl group, the substituent-containing compound may be the halogen-containing compound N-halosuccinimide, and the reactive psilocybin derivative and the substituent-containing compound may react to form a first psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be a methoxycarbonyl group and at least one of R2, R4, R5, R6, or R7 may be a halogen atom.

[0068] In at least one embodiment of one aspect, the N-halosuccinimide may be N-chlorosuccinimide, and the first psilocybin derivative formed may be the compound shown in Figure 13B and representation 13B-3.

[0069] In at least one embodiment of one aspect, the substituent of the reactive psilocybin derivative of formula (II) may be a methoxycarbonyl group, the substituent-containing compound may be the nitro-containing compound nitronium tetrafluoroborate, and the reactive psilocybin derivative and the substituent-containing compound may react to form a second psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 is a methoxycarbonyl group and at least one of R2, R4, R5, R6, or R7 is a nitro group.

[0070] In at least one embodiment of one aspect, the second psilocybin derivative formed may be the compound shown in Figure 13B and designation 13B-4.

[0071] In at least one embodiment of one aspect, the formed first psilocybin derivative of formula (I) may be reacted with an acetylated glycosyl compound to form a third psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be a methoxycarbonyl group, at least one of R2, R4, R5, R6, or R7 may be a glycosyloxy group, and at least one of R2, R4, R5, R6, or R7 may be a halogen atom.

[0072] In at least one embodiment of one aspect, the third psilocybin derivative formed may be the compound shown in Figure 13B and designation 13B-5.

[0073] In at least one embodiment of one aspect, the formed second psilocybin derivative of formula (I) may be reacted with an acetylated glycosyl compound to form a fourth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be a methoxycarbonyl group, at least one of R2, R4, R5, R6, or R7 may be a glycosyloxy group, and at least one of R2, R4, R5, R6, or R7 is a nitro group.

[0074] In at least one embodiment of one aspect, the quaternary psilocybin derivative formed may be the compound shown in Figure 13B and designation 13B-6.

[0075] In at least one embodiment of one aspect, the formed third psilocybin derivative of formula (I) may be reacted with ammonia to convert the methoxycarbonyl group of the amide group to form a fifth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be an amide group, at least one of R2, R4, R5, R6, or R7 may be a glycosyloxy group, and at least one of R2, R4, R5, R6, or R7 may be a halogen atom.

[0076] In at least one embodiment of one aspect, the fifth psilocybin derivative formed may be the compound shown in Figure 13B and designation 13B-7.

[0077] In at least one embodiment of one aspect, the formed fourth psilocybin derivative of formula (I) may be reacted with ammonia to convert the methoxycarbonyl group of the amide group to form a sixth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be an amide group, at least one of R2, R4, R5, R6, or R7 may be a glycosyloxy group, and at least one of R2, R4, R5, R6, or R7 is a nitro group.

[0078] In at least one embodiment of one aspect, the sixth psilocybin derivative formed may be the compound shown in Figure 13B and designation 13B-8.

[0079] In at least one embodiment of one aspect, the sixth psilocybin derivative of formula (I) formed may react to reduce the nitro group to form an amino group and a seventh psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be an amide group, at least one of R2, R4, R5, R6, or R7 may be a glycosyloxy group, and at least one of R2, R4, R5, R6, or R7 may be a nitro group.

[0080] In at least one embodiment of one aspect, the seventh psilocybin derivative formed may be the compound shown in Figure 13B and designation 13B-9.

[0081] In at least one embodiment of one aspect, the substituent of the reactive psilocybin derivative of formula (II) may be an acetamidyl group, the substituent-containing compound may be the halogen-containing compound N-halosuccinimide, and the reactive psilocybin derivative and the substituent-containing compound may react to form a first psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be an acetamidyl group and at least one of R2, R4, R5, R6, or R7 may be a halogen atom.

[0082] In at least one embodiment of one aspect, the N-halosuccinimide may be N-bromosuccinimide (NBS), in which case the first psilocybin derivative formed may be the compound shown in Figure 13C and representation 13C-6.

[0083] In at least one embodiment of one aspect, the substituent of the reactive psilocybin derivative of Formula (II) may be an acetamidyl group, the substituent-containing compound may be dimethylformamide, and the reactive psilocybin derivative and the substituent-containing compound may react to form an intermediate psilocybin derivative of Formula (I), in which one of R2, R4, R5, R6, or R7 may be an acetamidyl group and at least one of R2, R4, R5, R6, or R77 may be a methanol group, and the intermediate psilocybin derivative may be further reacted to oxidize the methanol group to form a second psilocybin derivative of Formula (I), in which one of R2, R4, R5, R6, or R7 may be an acetamidyl group and at least one of R2, R4, R5, R6, or R7 may be a carboxy group.

[0084] In at least one embodiment of one aspect, the intermediate psilocybin derivative may be the compound shown in Figure 13C and representation 13C-5, and the second psilocybin derivative formed may be the compound shown in Figure 13C and representation 13C-7.

[0085] In at least one embodiment of one aspect, the formed second psilocybin derivative of formula (I) may be reacted with an alcohol to esterify the carboxy group to form an ester and a third psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be an acetamidyl group and at least one of R2, R4, R5, R6, or R7 may be a carboxyl ester.

[0086] In at least one embodiment of one aspect, the third psilocybin derivative formed may be the compound shown in Figure 13C and designation 13C-8.

[0087] In at least one embodiment of one aspect, the formed third psilocybin derivative of formula (I) may be reacted with a nitro-group-containing compound to form a fourth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be an acetamidyl group, at least one of R2, R4, R5, R6, or R7 may be a carboxyl ester, and at least one of R2, R4, R5, R6, or R7 is a nitro group.

[0088] In at least one embodiment of one aspect, the quaternary psilocybin derivative formed may be the compound shown in Figure 13C and designation 13C-9.

[0089] In at least one embodiment of one aspect, the fourth psilocybin derivative of formula (I) formed may undergo a reaction to reduce the nitro group to form an amino group and a fifth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 is an acetamidyl group, at least one of R2, R4, R5, R6, or R7 is a carboxyl ester, and at least one of R2, R4, R5, R6, or R7 is an amino group.

[0090] In at least one embodiment of one aspect, the fifth psilocybin derivative formed may be the compound shown in Figure 13C and designation 13C-10.

[0091] In at least one embodiment of one aspect, the formed fifth psilocybin derivative of formula (I) may react with ammonia to form an amide group and a sixth psilocybin derivative of formula (I), wherein one of R2, R4, R5, R6, or R7 may be an acetamidyl group, wherein at least one of R2, R4, R5, R6, or R7 is a carboxyl ester and at least one of R2, R4, R5, R6, or R7 may be an amide group.

[0092] In at least one embodiment of one aspect, the sixth psilocybin derivative formed may be the compound shown in Figure 13C and designation 13C-11.

[0093] In another aspect, the present disclosure relates to further methods of forming poly-substituted psilocybin derivatives. Accordingly, in one aspect, the present disclosure provides, in at least one embodiment, a method of making a poly-substituted psilocybin derivative, i.e., a compound represented by the following formula (LVII): [ka] wherein at least one of R2, R4, R5, R6, or R7 is a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and wherein each unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R3 is a hydrogen atom, -CH2-CHNH2COOH, or -CH2-CH2NH2, contacting a psilocybin derivative precursor compound of formula (I) with a catalytic amount of a psilocybin biosynthetic enzyme complement under reaction conditions that allow for enzyme-catalyzed conversion of the psilocybin derivative precursor compound to a compound of formula (I): [ka] (wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and when each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom, and R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group).

[0094] In at least one embodiment of one aspect, the reaction conditions may be in vitro reaction conditions.

[0095] In at least one embodiment of one aspect, the reaction conditions may be in vivo reaction conditions.

[0096] In at least one embodiment of one aspect, the psilocybin derivative precursor compound and the substituent-containing compound may be contacted with a psilocybin biosynthetic enzyme complement in a host cell, the host cell containing, as operably linked components: (i) a nucleic acid sequence that controls expression in a host cell; and (ii) The host cell may have a chimeric nucleic acid sequence that includes a nucleic acid sequence encoding a psilocybin biosynthetic enzyme complement, and the host cell may be grown to express the psilocybin biosynthetic enzyme complement and produce a multi-substituted psilocybin derivative compound.

[0097] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, and SEQ ID NO:48, (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, and SEQ ID NO:49; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, and SEQ ID NO:49; and (g) may have at least one enzyme encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0098] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) a tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f), wherein in the psilocybin derivative precursor compound of formula (LVII), two of R2, R4, R5, R6, or R7 are substituents independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom, thereby forming a first polysubstituted psilocybin derivative compound of formula (I), and R 3c is a carboxyl group.

[0099] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LVIII): [ka] wherein R4 is a hydroxy group and R6 is a chlorine atom, and the first polysubstituted psilocybin derivative compound formed has the following formula (LV): [ka] It has.

[0100] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement decarboxylates the R—CH—CHNH—COOH group to form a compound of formula (I) (R 3a and R 3b and each is a hydrogen atom), the tryptophan decarboxylase being capable of reacting with the following: (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0101] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LVIII): [ka] (wherein R4 is a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom), and the first polysubstituted psilocybin derivative compound formed may have the following formula (LIX): [ka] wherein R4 is a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom, and the second polysubstituted psilocybin derivative has the following formula (XXII), (XXVI), (XXIX), (LII), or (LIV): [ka] It has the following characteristics.

[0102] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is represented by formula (I) (R3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0103] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LVIII): [ka] wherein R4 is an acetamidyl group, a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom, and the first polysubstituted psilocybin derivative compound formed has the following formula (LIX): [ka] wherein R4 is an acetamidyl group, a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom, and the second polysubstituted psilocybin derivative has the following formula (LX): [ka] and the third polysubstituted psilocybin derivative has the following formula (IX), (X), (XVIII), (XXI), (XXV), or (XXVIII): [ka] It has the following characteristics.

[0104] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement methylates the R3 amino group of R3 to form a compound of formula (I) (R 3a and R 3b are methyl groups, or R 3a is a hydrogen atom, and R 3b is a methyl group), and the N-methyltransferase may further comprise an N-methyltransferase that forms a fourth polysubstituted psilocybin derivative (wherein (a) SEQ ID NO: 11 and SEQ ID NO: 13; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0105] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LVIII): [ka] (wherein R4 is an amino group or a hydroxy group, and R6 is a chlorine atom, a nitrile group, or a bromine atom), and the first polysubstituted psilocybin derivative compound formed may have the following formula (LIX): [ka] wherein R4 is an amino group or a hydroxy group, and R6 is a chlorine atom, a nitrile group, or a bromine atom, and the second polysubstituted psilocybin derivative has the following formula (LX): [ka] and the quaternary polysubstituted psilocybin derivatives have the following formula (XXVII), (XL), or (LIII): [ka] It has the following characteristics.

[0106] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, and SEQ ID NO: 21; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, and SEQ ID NO: 22; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, and SEQ ID NO: 22; and (g) a prenyltransferase encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f), wherein in the psilocybin derivative precursor compound of formula (LVII), one of R2, R4, R5, R6, or R7 is a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is -CH2-CHNH2COOH, forming a first polysubstituted psilocybin derivative compound of formula (I), wherein at least one of R2, R4, R5, R6, or R7 is a prenyl group and R 3c is a halogen atom.

[0107] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXI): [ka] wherein R4 is a hydroxy group, and the first polysubstituted psilocybin derivative formed has the following formula (L): [ka] It has.

[0108] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement decarboxylates the R—CH—CHNH—COOH group to form a compound of formula (I) (R 3a and R 3b and each are a hydrogen atom), wherein the tryptophan decarboxylase is (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0109] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement methylates the R3 amino group of R3 to form a compound of formula (I) (R 3a and R 3b are methyl groups, or R 3a is a hydrogen atom, and R 3b is a methyl group), and the N-methyltransferase may further comprise an N-methyltransferase that forms a fourth polysubstituted psilocybin derivative (wherein (a) SEQ ID NO: 11 and SEQ ID NO: 13; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0110] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXI): [ka] wherein R4 is a propionyloxy group or an acetoxy group, and the first polysubstituted psilocybin derivative compound formed has the following formula (LIX): [ka] wherein R4 is a propionyloxy group or an acetoxy group, and R6 is a prenyl group, and the second polysubstituted psilocybin derivative has the formula: [ka] wherein R4 is a propionyloxy group or an acetoxy group, and R6 is a prenyl group, and the third polysubstituted psilocybin derivative has the following formula (XLI) or (XLII): [ka] It has the following characteristics.

[0111] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXII): [ka] wherein R5 is a chlorine or fluorine atom, and the first polysubstituted psilocybin derivative compound formed has the following formula (LXIII): [ka] wherein R5 is a chlorine or fluorine atom and R6 is a prenyl group, and the second polysubstituted psilocybin derivative formed has the following formula (XXXVI) or (XXXVIII): [ka] It has the following characteristics.

[0112] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is represented by formula (I) (R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0113] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXI): [ka] wherein R5 is a chlorine or fluorine atom, and the first polysubstituted psilocybin derivative compound formed has the following formula (LXIII): [ka] wherein R5 is a chlorine or fluorine atom and R6 is a prenyl group, and the second polysubstituted psilocybin derivative formed has the following formula (LXIV): [ka] wherein R5 is a chlorine or fluorine atom and R6 is a prenyl group, and the third polysubstituted psilocybin derivative formed has the following formula (XXXV) or (XXXVII): [ka] It has the following characteristics.

[0114] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) a tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f), wherein in the psilocybin derivative precursor compound of formula (LVIII), two of R2, R4, R5, R6, or R7 are substituents independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom, thereby forming a first polysubstituted psilocybin derivative compound of formula (I), and R 3c is a carboxyl group.

[0115] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXV): [ka] wherein R4 is a hydroxy group and R5 is a prenyl group, and the first polysubstituted psilocybin derivative compound formed has the following formula (LI): [ka] It has.

[0116] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement decarboxylates the R—CH—CHNH—COOH group to form a compound of formula (I) (R 3a and R 3b and each are a hydrogen atom), wherein the tryptophan decarboxylase is (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0117] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXV): [ka] (wherein R4 is a fluorine atom and R5 is a nitrile group), and the first polysubstituted psilocybin derivative compound formed may be a compound of the following formula (LXIX): [ka] wherein R4 is a fluorine atom and R5 is a nitrile group, and the second polysubstituted psilocybin derivative formed has the following formula (XIX): [ka] It has the following characteristics.

[0118] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is represented by formula (I) (R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0119] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXV): [ka] (wherein R4 is a fluorine atom and R5 is a hydroxy group or a nitrile group), and the first polysubstituted psilocybin derivative compound formed may be a compound of the following formula (LXIX): [ka] wherein R4 is a fluorine atom and R5 is a hydroxy group or a nitrile group, and the second polysubstituted psilocybin derivative formed has the following formula (LXXII): [ka] wherein R4 is a fluorine atom and R5 is a hydroxy group or a nitrile group, and the third polysubstituted psilocybin derivative formed has the following formula (XVII) or (XX): [ka] It has the following characteristics.

[0120] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) a tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f), wherein in the psilocybin derivative precursor compound of formula (LVIII), two of R2, R4, R5, R6, or R7 are substituents independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom, thereby forming a first polysubstituted psilocybin derivative compound of formula (I), and R 3c is a carboxyl group.

[0121] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVI): [ka] wherein R4 is a hydroxy group and R7 is a prenyl group, and the first polysubstituted psilocybin derivative compound formed has the following formula (XLIX): [ka] It has.

[0122] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement decarboxylates the R—CH—CHNH—COOH group to form a compound of formula (I) (R 3a and R 3b and each are a hydrogen atom), wherein the tryptophan decarboxylase is (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0123] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVI): [ka] (wherein R4 is a fluorine atom and R7 is a nitrile group), and the first polysubstituted psilocybin derivative compound formed may be a compound of the following formula (LXX): [ka] (wherein R4 is a fluorine atom and R7 is a nitrile group), and the second polysubstituted psilocybin derivative compound formed has the following formula (XXIV): [ka] It has.

[0124] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is represented by formula (I) (R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0125] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVI): [ka] (wherein R4 is a fluorine atom or a chlorine atom, and R7 is a prenyl group or a nitrile group), and the first polysubstituted psilocybin derivative compound formed may have the following formula (LXX): [ka] (wherein R4 is a fluorine atom or a chlorine atom, and R7 is a prenyl group or a nitrile group), and the second polysubstituted psilocybin derivative compound formed has the following formula (LXXIII): [ka] wherein R4 is a fluorine atom or a chlorine atom, and R7 is a prenyl group or a nitrile group, and the third polysubstituted psilocybin derivative compound formed has the following formula (XXIII) or (XX): [ka] It has.

[0126] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement methylates the R3 amino group of R3 to form a compound of formula (I) (R 3a and R 3b are methyl groups, or R 3a is a hydrogen atom, and R 3b is a methyl group), and the N-methyltransferase may further comprise an N-methyltransferase that forms a fourth polysubstituted psilocybin derivative (wherein (a) SEQ ID NO: 11 and SEQ ID NO: 13; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0127] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVI): [ka] (wherein R4 is a chlorine atom and R7 is a hydroxy group), and the first polysubstituted psilocybin derivative compound formed may have the following formula (LXX): [ka] (wherein R4 is a hydroxy group and R7 is a chlorine atom), and the second polysubstituted psilocybin derivative compound formed has the following formula (LXXIII): [ka] wherein R4 is a hydroxy group and R7 is a chlorine atom, and the third polysubstituted psilocybin derivative compound formed has the following formula (XXXIX): [ka] It has.

[0128] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) a tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from the nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f), wherein in the psilocybin derivative precursor compound of formula (LVIII), any two of R2, R4, R5, R6, or R7 are selected from the group consisting of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group, and (ix) a nitrile group, wherein R is a hydrogen atom, and a first polysubstituted psilocybin derivative compound of formula (I) is formed, wherein R is a carboxyl group, and the psilocybin biosynthetic enzyme complement removes the carboxyl group from the R-CH-CHNHCOOH group of the first polysubstituted psilocybin derivative compound to form a compound of formula (I) (R 3a and R 3band each is a hydrogen atom), wherein the tryptophan decarboxylase forms a second polysubstituted psilocybin derivative of: (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0129] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVII): [ka] (wherein R5 is a fluorine atom, a chlorine atom, or a nitrile group, and R6 is a fluorine atom, an amino group, or a prenyl group), and the first polysubstituted psilocybin derivative compound formed may be a compound of the following formula (LXIII): [ka] (wherein R5 is a fluorine atom, a chlorine atom, or a nitrile group, and R6 is a fluorine atom, an amino group, or a prenyl group), and the second polysubstituted psilocybin derivative compound formed has the following formula (XI), (XVI), (XXXVI), or (XXXVIII): [ka] It has.

[0130] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is represented by formula (I) (R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0131] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVII): [ka] (wherein R5 is a fluorine atom or a chlorine atom, and R6 is an amino group, an acetamidyl group, or a prenyl group), and the first polysubstituted psilocybin derivative compound formed may be a compound of the following formula (LXIII): [ka] (wherein R5 is a fluorine atom or a chlorine atom, and R6 is an amino group, an acetamidyl group, or a prenyl group), and the second polysubstituted psilocybin derivative compound formed has the following formula (LXXIV): [ka] wherein R5 is a fluorine atom or a chlorine atom, and R6 is an amino group, an acetamidyl group, or a prenyl group, and the third polysubstituted psilocybin derivative compound formed has the following formula (XIV), (XV), (XXXV), or (XXXVII): [ka] It has.

[0132] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement methylates the R3 amino group of R3 to form a compound of formula (I) (R 3a and R 3b are methyl groups, or R 3a is a hydrogen atom, and R 3b is a methyl group), and the N-methyltransferase may further comprise an N-methyltransferase that forms a fourth polysubstituted psilocybin derivative (wherein (a) SEQ ID NO: 11 and SEQ ID NO: 13; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0133] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVII): [ka] (wherein R5 is a chlorine atom and R6 is a prenyl group), and the first polysubstituted psilocybin derivative compound formed may have the following formula (LXIII): [ka] (wherein R5 is a chlorine atom and R6 is a prenyl group), and the second polysubstituted psilocybin derivative compound formed has the following formula (LXXIV): [ka] (wherein R5 is a chlorine atom and R6 is a prenyl group), and the third polysubstituted psilocybin derivative compound formed has the following formula (XLIV): [ka] It has.

[0134] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) a tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f); wherein in the psilocybin derivative precursor compound of formula (LVIII), two of R2, R4, R5, R6, or R7 are substituents independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom, thereby forming a first polysubstituted psilocybin derivative compound of formula (I), and R 3c is a carboxyl group, and the psilocybin biosynthetic enzyme complement decarboxylates the R-CH-CHNHCOOH group of the first polysubstituted psilocybin derivative compound to form a compound represented by formula (I) (R 3a and R 3b and each is a hydrogen atom), wherein the tryptophan decarboxylase forms a second polysubstituted psilocybin derivative of: (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0135] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVIII): [ka] (wherein R5 is a fluorine atom and R7 is a nitro group or a prenyl group), and the first polysubstituted psilocybin derivative compound formed may have the following formula (LXXI): [ka] wherein R5 is a fluorine atom and R7 is a nitro group atom or a prenyl group, and the second polysubstituted psilocybin derivative compound formed has the following formula (XIII) or (XXXIII): [ka] It has.

[0136] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is represented by formula (I) (R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0137] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXVIII): [ka] (wherein R5 is a fluorine atom and R7 is a nitro group or a prenyl group), and the first polysubstituted psilocybin derivative compound formed may have the following formula (LXXI): [ka] (wherein R5 is a fluorine atom and R7 is a nitro group atom or a prenyl group), and the second polysubstituted psilocybin derivative compound formed has the following formula (LXXV): [ka] wherein R5 is a fluorine atom and R7 is a nitro group or a prenyl group, and the third polysubstituted psilocybin derivative compound formed has the following formula (XII) or (XXXII): [ka] It has.

[0138] In at least one embodiment of one aspect, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, and SEQ ID NO: 21; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, and SEQ ID NO: 22; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, and SEQ ID NO: 22; and (g) a prenyltransferase encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f), wherein in the psilocybin derivative precursor compound of formula (LVIII), one of R2, R4, R5, R6, or R7 is a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom, such that a first polysubstituted psilocybin derivative compound of formula (I) is formed; 3c is a carboxy group or a halogen atom.

[0139] In at least one embodiment of one aspect, the psilocybin derivative precursor compound has the following formula (LXXVII): [ka] and the first polysubstituted psilocybin derivative compound has the following formula (LXXVI): [ka] It has.

[0140] In at least one embodiment of one aspect, the method may further comprise isolating the multi-substituted psilocybin derivative compound from the host cell and / or host cell medium.

[0141] In at least one embodiment of one aspect, the host cell may be a microorganism.

[0142] In at least one embodiment of one aspect, the host cell may be a bacterial cell or a yeast cell.

[0143] In at least one embodiment of one aspect, the host cell may be Escherichia coli or Saccharomyces cerevisiae.

[0144] In at least one embodiment of another aspect, the present disclosure provides a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group) or a salt thereof in the manufacture of a pharmaceutical or recreational drug formulation.

[0145] In at least one embodiment, the manufacturing can include formulating the compound with an excipient, diluent, or carrier.

[0146] In at least one embodiment of another aspect, the present disclosure provides a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group) or a salt thereof, is provided for use as a pharmaceutical or recreational drug formulation, together with an excipient, diluent, or carrier.

[0147] Other features and advantages will become apparent from the following detailed description, which, however, while indicating preferred embodiments of the present disclosure, is given by way of illustration only, since various changes and modifications within the spirit and scope of the present disclosure will become apparent to those skilled in the art from said detailed description. [Brief explanation of the drawings]

[0148] The present disclosure will be described, by way of example, in the sections provided below with reference to the accompanying drawings. The drawings provided herein are provided for a better understanding of the exemplary embodiments and also to more clearly show how various embodiments may be practiced. Such drawings are not intended to limit the present disclosure in any way.

[0149] [Figure 1] Figure 1 shows the chemical structure of psilocybin. [Figure 2]Figure 2 shows a specific prototypical structure for psilocybin and psilocybin derivatives, i.e., indole. Specific carbon and nitrogen atoms are referred to by reference to their position within the indole structure (i.e., N1, C2, C3, etc.). Appropriate atomic numbers are indicated. [Figures 3A-3L] 3A, 3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I, 3J, 3K, and 3L show the chemical structures of certain exemplary psilocybin derivatives, in particular, 2,5-di-S1,S2-psilocybin derivatives (FIG. 3A), 2,5-di-S2,S1-psilocybin derivatives (FIG. 3B), 2,6-di-S1,S2-psilocybin derivatives (FIG. 3C), 2,6-di-S2,S1-psilocybin derivatives (FIG. 3D), and 2,7-di-S1,S2-psilocybin derivatives. (Figure 3E), 2,7-di-S2,S1-psilocybin derivatives (Figure 3F), 5,6-di-S1,S2-psilocybin derivatives (Figure 3G), 5,6-di-S2,S1-psilocybin derivatives (Figure 3H), 5,7-di-S1,S2-psilocybin derivatives (Figure 3I), 5,7-di-S2,S1-psilocybin derivatives (Figure 3J), 6,7-di-S1,S2-psilocybin derivatives (Figure 3K), and 6,7-di-S2,S1-psilocybin derivatives (Figure 3L). Each of S1 and S2 above is selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of Figures 3A to 3L, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. [Figures 4A-4F]Figures 4A, 4B, 4C, 4D, 4E, and 4F show the chemical structures of certain exemplary psilocybin derivatives, in particular, a 2,5,6-tri-S1,S2,S3-psilocybin derivative (Figure 4A), a 2,5,6-tri-S1,S3,S2-psilocybin derivative (Figure 4B), a 2,5,6-tri-S3,S1,S2-psilocybin derivative (Figure 4C), a 2,5,6-tri-S2,S1,S3-psilocybin derivative (Figure 4D), a 2,5,6-tri-S3,S2,S1-psilocybin derivative (Figure 4E), and a 2,5,6-tri-S2,S3,S1-psilocybin derivative (Figure 4F). Each of S1, S2, and S3 is selected from the group consisting of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of Figures 4A to 4F, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. Figures 4G, 4H, and 4I show the chemical structures of certain exemplary psilocybin derivatives, specifically, the 2,5,6-tri-S1,S2,S2-psilocybin derivative (Figure 4G), the 2,5,6-tri-S2,S1,S2-psilocybin derivative (Figure 4H), and the 2,5,6-tri-S2,S2,S1-psilocybin derivative (Figure 4I). Each of S1 and S2 is selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of FIGS. 4G to 4I, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. [Figures 5A-5F]Figures 5A, 5B, 5C, 5D, 5E, and 5F show the chemical structures of certain exemplary psilocybin derivatives, in particular, a 2,5,7-tri-S1,S2,S3-psilocybin derivative (Figure 5A), a 2,5,7-tri-S1,S3,S2-psilocybin derivative (Figure 5B), a 2,5,7-tri-S3,S1,S2-psilocybin derivative (Figure 5C), a 2,5,7-tri-S2,S1,S3-psilocybin derivative (Figure 5D), a 2,5,7-tri-S3,S2,S1-psilocybin derivative (Figure 5E), and a 2,5,7-tri-S2,S3,S1-psilocybin derivative (Figure 5F). Each of S1, S2, and S3 is selected from the group consisting of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of Figures 5A-5F, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. Figures 5G, 5H, and 5I show the chemical structures of certain exemplary psilocybin derivatives, particularly the 2,5,7-tri-S1,S2,S2-psilocybin derivative (Figure 5G), the 2,5,7-tri-S2,S1,S2-psilocybin derivative (Figure 5H), and the 2,5,7-tri-S2,S2,S1-psilocybin derivative (Figure 5I). Each of S1 and S2 is selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of FIGS. 5A to 5I, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. [Figures 6A-6F]Figures 6A, 6B, 6C, 6D, 6E, and 6F show the chemical structures of certain exemplary psilocybin derivatives, in particular, a 2,6,7-tri-S1,S2,S3-psilocybin derivative (Figure 6A), a 2,6,7-tri-S1,S3,S2-psilocybin derivative (Figure 6B), a 2,6,7-tri-S3,S1,S2-psilocybin derivative (Figure 6C), a 2,6,7-tri-S2,S1,S3-psilocybin derivative (Figure 6D), a 2,6,7-tri-S3,S2,S1-psilocybin derivative (Figure 6E), and a 2,6,7-tri-S2,S3,S1-psilocybin derivative (Figure 6F). Each of S1, S2, and S3 is selected from the group consisting of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of Figures 6A to 6F, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. Figures 6G, 6H, and 6I show the chemical structures of certain exemplary psilocybin derivatives, particularly the 2,6,7-tri-S1,S2,S2-psilocybin derivative (Figure 6G), the 2,6,7-tri-S2,S1,S2-psilocybin derivative (Figure 6H), and the 2,6,7-tri-S2,S2,S1-psilocybin derivative (Figure 6I). Each of S1 and S2 is selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of FIGS. 6A to 6F, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. [Figures 7A-7F]Figures 7A, 7B, 7C, 7D, 7E, and 7F show the chemical structures of certain exemplary psilocybin derivatives, in particular, a 5,6,7-tri-S1,S2,S3-psilocybin derivative (Figure 7A), a 5,6,7-tri-S1,S3,S2-psilocybin derivative (Figure 7B), a 5,6,7-tri-S3,S1,S2-psilocybin derivative (Figure 7C), a 5,6,7-tri-S2,S1,S3-psilocybin derivative (Figure 7D), a 5,6,7-tri-S3,S2,S1-psilocybin derivative (Figure 7E), and a 5,6,7-tri-S2,S3,S1-psilocybin derivative (Figure 7F). Each of S1, S2, and S3 is selected from the group consisting of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of Figures 7A-7F, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. Figures 7G, ​​7H, and 7I show the chemical structures of certain exemplary psilocybin derivatives, specifically, the 5,6,7-tri-S1,S2,S2-psilocybin derivative (Figure 7G), the 5,6,7-tri-S2,S1,S2-psilocybin derivative (Figure 7H), and the 5,6,7-tri-S2,S2,S1-psilocybin derivative (Figure 7I). Each of S1 and S2 is selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. In each of FIGS. 7A to 7F, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. [Figures 8A-8G]Figures 8A, 8B, 8C, 8D, 8E, 8F, and 8G show the chemical structures of certain exemplary psilocybin derivatives, particularly O-alkylated psilocybin derivatives, specifically 4-O-methyl-5-chloropsilocybin derivatives (Figure 8A), 4-O-ethyl-5-chloropsilocybin derivatives (Figure 8B), 4-acetoxy-5-chloropsilocybin derivatives (Figure 8C), 4-propionyloxy-5-chloropsilocybin derivatives (Figure 8D), 4-hydroxy-5-chloropsilocybin derivatives (Figure 8E), 4-phospho-5-chloropsilocybin derivatives (Figure 8F), and 5-chloropsilocybin derivatives (Figure 8H). In each of Figures 8A-8G, R3a and R3b may each be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R3c may be a hydrogen atom or a carboxyl group. [Figure 9] FIG. 9 illustrates an exemplary chemical reaction for synthesizing nitrated psilocybin derivatives, specifically, reacting a 4-O-methyl psilocybin derivative with nitric acid in the presence of sulfuric acid to form a 4-O-methyl-5-nitro psilocybin derivative. [Figure 10] FIG. 10 shows exemplary chemical reactions for synthesizing glycosylated psilocybin derivatives, specifically, the reaction of a 4-hydroxypsilocybin derivative with a glycosyl compound to form a 4-glycosylpsilocybin derivative. [Figures 11A-11C] Figures 11A, 11B, and 11C show specific exemplary chemical reactions for synthesizing aminated psilocybin derivatives and the resulting N-substituted derivatives, specifically the reaction of 4-O-methyl-5-nitropsilocybin derivatives with hydrogen under palladium-on-charcoal catalysis to form 4-O-methyl-5-aminopsilocybin derivatives. The amino group formed at the 5-position can be substituted with various groups, such as acylation using acetic anhydride. The amino group can also be alkylated by condensation with an aldehyde (e.g., acetaldehyde) followed by reduction of the intermediate imine with borohydride (Figure 11B). Figure 11C shows a possible direct amination using HO and NH HO under catalysis. [Figures 12A-12D]Figures 12A, 12B, 12C, and 12D show exemplary chemical reactions, particularly the formation of a 4-hydroxy-5-carboxyl psilocybin derivative using a 4-hydroxy psilocybin derivative as a reactant (Figure 12A), the formation of a 4-hydroxy-7-carboxyl psilocybin derivative using an aryl halide psilocybin derivative as a reactant (Figure 12B), the formation of a carboxylated protected psilocybin derivative using a protected psilocybin derivative as a reactant (Figure 12C), and the formation of a sodium salt of a carboxylated psilocybin derivative (a), an ester via an OH-substituted carboxyl group (b), or an amide via an OH-substituted carboxyl group (c) using a carboxylated psilocybin derivative as a reactant (Figure 12D). [Figures 13A-13D]Figures 13A, 13B, 13C, and 13D show exemplary chemical reactions for forming psilocybin derivatives. Figure 13A shows the conversion of the initially synthesized 7-nitrated psilocybin derivative into another psilocybin derivative having two types of groups at the C5 and C7 atoms, where the C5 atom is a keto or carboxyl group and the C7 atom is nitro, amine, nitrile, hydroxyl, iodine, or fluorine, among others. Figure 13B shows the conversion of the initially synthesized 5-carboxy psilocybin derivative into another O-4-glycosylated psilocybin derivative having two types of groups at the C5 and C7 atoms, where the C5 atom is an ester or amide and the C7 atom is nitro or amine, among others. Figure 13C shows a further exemplary chemical conversion of the initially synthesized 5-nitrated psilocybin derivative into a 5,6,7 trisubstituted psilocybin derivative containing up to three types of groups at the C5, C6, and C7 carbon atoms. The group attached to the C5 carbon atom is a nitro, amino, or N-acetamido group; the group attached to the C6 carbon atom is a halide, nitro, or amino group; and the group attached to the C7 carbon atom is a formyl, carboxy, or amido group. Figure 13D shows the preparation of a hydroxypsilocybin derivative (Figure 13-D4), which may then be prenylated in vitro or in vivo using a prenyltransferase to form, for example, a C6 prenylated derivative of compound 13-D4. Note that specific compounds shown in Figures 13A-13D are labeled 13A-1, 13A-2, etc. (Figure 13A), 13B-1, 13B-2, etc. (Figure 13B), 13C-1, 13C-2, etc. (Figure 13C), and 13D-1, 13D-2, etc. (Figure 13D). [Figures 14A-14G]Figures 14A, 14B, 14C, 14D, 14E, 14F, and 14G illustrate specific routes for biochemically generating exemplary polysubstituted psilocybin derivative compounds, including, in particular, an exemplary compound having an S14 and S26 substituent (Figure 14A), an exemplary compound having an S14 and S26 prenyl group (Figure 14B), an exemplary compound having an S15 and S26 prenyl group (Figure 14C), an exemplary compound having an S14 and S25 substituent (Figure 14D), an exemplary compound having an S14 and S27 substituent (Figure 14E), an exemplary compound having an S15 and S26 substituent (Figure 14F), and an exemplary compound having an S15 and S27 substituent (Figure 14G). The Roman numerals adjacent to each exemplary compound in Figures 14A-14G refer to the exemplary compound so numbered in this disclosure. [Figures 15A-15H]15A, 15B, 15C, 15D, 15E, 15F, 15G, and 15H show various graphs obtained in experimental assays to evaluate the effects of exemplary prenylated psilocybin derivatives having formula (IX) described herein, in particular a cell viability assay for a multi-substituted psilocybin derivative having formula (IX) (FIG. 15A); a saturation binding assay for [H]ketanserin at the 5-HT receptor (FIG. 15B); a competition assay for thyroxine as a positive control (binding) (Panel A) and a competition assay for tryptophan as a negative control (non-binding) (Panel B) (FIG. 15C); a competition assay for a multi-substituted psilocybin derivative compound having formula IX (referred to as "IX") (FIG. 15D, which for clarity is shown on two different axes, i.e., Panels A and B). 15A), a cAMP assay with increasing concentrations of psilocin in +5HT1A and -5HT1A cells in the presence of a constant (4 μM) forskolin (Figure 15E), a cAMP assay with increasing concentrations of serotonin +5HT1A and -5HT1A cells in the presence of a constant (4 μM) forskolin and 10 μM serotonin (Figure 15F), a cAMP assay with increasing concentrations of tryptophan in +5HT1A and -5HT1A cells in the presence of a constant (4 μM) forskolin (Figure 15G), and a cAMP assay with increasing concentrations of a multi-substituted psilocybin derivative compound having formula IX (referred to as "IX") in +5HT1A and -5HT1A cells in the presence of a constant (4 μM) forskolin (Figure 15H). [Figures 16A-16C] Figures 16A, 16B, and 16C show various graphs obtained in experimental assays to evaluate the effects of exemplary prenylated psilocybin derivatives having formula (X) described herein, in particular a cell viability assay for a multi-substituted psilocybin derivative having formula (IX) (Figure 16A), a competition assay for a multi-substituted psilocybin derivative compound having formula IX (designated "IX") (Figure 16B), and a cAMP assay with increasing concentrations of a multi-substituted psilocybin derivative compound having formula X (designated "X") in +5HT1A and -5HT1A cells in the presence of a constant (4 μM) forskolin (Figure 16H). [Figures 17A-17B] 17A and 17B show chromatograms, and in particular mass spectrometry data in the form of a chromatogram (FIG. 17A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the formula (XII) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (FIG. 17B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the formula (XII) described herein. [Figures 18A-18B] Figures 18A and 18B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 18A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XIV) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 18B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XII) described herein. [Figures 19A-19B] Figures 19A and 19B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 19A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XV) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 19B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XV) described herein. [Figure 20] FIG. 20 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XVII) described herein. [Figures 21A-21B] Figures 21A and 21B show chromatograms, and in particular further mass spectrometry data in the form of a chromatogram (Figure 21A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XVIII) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 21B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XVIII) described herein. [Figures 22A-22B]Figures 22A and 22B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 22A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXI) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 22B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXI) described herein. [Figures 23A-23B] Figures 23A and 23B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 23A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the formula (XXIII) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 23B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the formula (XXIII) described herein. [Figure 24] FIG. 24 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXV) described herein. [Figures 25A-25B] Figures 25A and 25B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 25A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXVIII) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 25B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXVIII) described herein. [Figure 26] FIG. 26 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XX) described herein. [Figure 27] FIG. 27 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XIII) described herein. [Figure 28] FIG. 28 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XVI) described herein. [Figure 29] FIG. 29 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XIX) described herein. [Figure 30] FIG. 30 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXII) described herein. [Figure 31] FIG. 31 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXVI) described herein. [Figure 32A-32B] Figures 32A and 32B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 32A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXIX) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 32B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXIX) described herein. [Figure 33] FIG. 33 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXVII) described herein. [Figure 34A-34B]Figures 34A and 34B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 34A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXXIV) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 34B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXXIV) described herein. [Figure 35A-35B] Figures 35A and 35B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 35A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXXII) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 35B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXXII) described herein. [Figure 36] FIG. 36 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXXVI) described herein. [Figure 37] FIG. 37 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XLIX) described herein. [Figure 38] FIG. 38 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (L) described herein. [Figure 39] FIG. 39 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XLVII) described herein. [Figure 40]FIG. 40 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (LI) described herein. [Figure 41] FIG. 41 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXXVI) described herein. [Figure 42] FIG. 42 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXXV) described herein. [Figure 43A-43B] Figures 43A and 43B show chromatograms, and in particular, further mass spectrometry data in the form of a chromatogram (Figure 43A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXXVI) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 43B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XXXVI) described herein. [Figure 44] FIG. 44 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXXVIII) described herein. [Figure 45] FIG. 45 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XL) described herein. [Figure 46] FIG. 46 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXXIX) described herein. [Figure 47]FIG. 47 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (LII) described herein. [Figure 48] FIG. 48 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (LIII) described herein. [Figure 49] FIG. 49 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (LIV) described herein. [Figure 50] FIG. 50 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXX) described herein. [Figure 51] FIG. 51 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XXXI) described herein. [Figure 52] FIG. 52 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XLI) described herein. [Figure 53] FIG. 53 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XLII) described herein. [Figure 54] Figure 54 shows chromatograms, and in particular additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having formula (XLV) described herein. [Figure 55A-55B]Figures 55A and 55B show chromatograms, particularly further mass spectrometry data in the form of a chromatogram (Figure 55A) obtained in an experiment to synthesize an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XLIV) described herein, and further mass spectrometry data in the form of a mass spectrometry spectrum (Figure 55B) obtained in an experiment to identify an exemplary poly-substituted psilocybin derivative compound having the chemical formula (XLIV) described herein. [Figure 56] Figure 56 shows chromatograms, and in particular, additional mass spectrometry data in the form of chromatograms, obtained in experiments to synthesize exemplary poly-substituted psilocybin derivative compounds having the formula (LXVVI) described herein.

[0150] The drawings, taken in conjunction with the following detailed description, will make apparent to those skilled in the art how the present disclosure may be practiced in practice. BEST MODE FOR CARRYING OUT THE INVENTION

[0151] Various compositions, systems, or processes are described below to provide example embodiments of each claimed subject matter. None of the embodiments described below limit the present subject matter, and the claimed subject matter may feature a different process, composition, or system than those described below. The claimed subject matter is not limited to compositions, processes, or systems having all or any one of the features of the compositions, systems, or processes described below, nor is it limited to features common to more than one or all of the compositions, systems, or processes described below. It is possible that a composition, system, or process described below is not an embodiment of the claimed subject matter. Subject matter disclosed in the compositions, systems, or processes described below but not claimed herein may be subject to other protection, such as a continuing patent application, in which the applicant(s), inventor(s), or owner(s) do not intend to abandon, disclaim, or dedicate to the public any such subject matter by disclosure herein.

[0152] In this specification and claims, the singular forms "a" or "an" and the definite article "the" include the plural forms, and vice versa, unless the context clearly dictates otherwise. Throughout this specification, unless otherwise specified, the terms "have," "include," and "having" are used inclusively rather than exclusively, such that a stated integer or group of integers includes one or more of the unstated integers or groups of integers.

[0153] Various compositions, systems, or processes are described below to provide example embodiments of each claimed subject matter. None of the embodiments described below limit the present subject matter, and the claimed subject matter may feature a different process, composition, or system than those described below. The claimed subject matter is not limited to compositions, processes, or systems having all or any one of the features of the compositions, systems, or processes described below, nor is it limited to features common to more than one or all of the compositions, systems, or processes described below. It is possible that a composition, system, or process described below is not an embodiment of the claimed subject matter. Subject matter disclosed in the compositions, systems, or processes described below but not claimed herein may be subject to other protection, such as a continuing patent application, in which the applicant(s), inventor(s), or owner(s) do not intend to abandon, disclaim, or dedicate to the public any such subject matter by disclosure herein.

[0154] When ranges are used herein with respect to physical properties, such as molecular weight, or chemical properties, such as chemical formulas, all combinations and subcombinations of ranges and specific embodiments therein are intended to be encompassed therein. Numerical values ​​expressing amounts of ingredients or reaction conditions used herein should be understood to be modified in all instances by the term "about," except in the operating examples or unless otherwise specified. The term "about," when referring to a numerical value or range of values, indicates that the stated numerical value or range of values ​​is an approximation within experimental variation (or within statistical experimental error); thus, the numerical value or range of values ​​may vary by 1% to 15% of the stated numerical value or range of values, as readily discernible depending on the context. Furthermore, ranges of values ​​described herein specifically include the limits of such ranges and any intermediate values ​​or subranges within a given range, and all such intermediate values ​​and subranges are intended to be individually and specifically disclosed (e.g., a range of 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.90, 4, and 5). Similarly, other terms of degree, such as "substantially" and "approximately," used herein mean reasonable deviations in the modifying term without significant variation in the end result. Such terms of degree should be construed to include deviations from the modifying term, unless such deviations negate the meaning of the modifying term.

[0155] Unless otherwise specified, pharmaceutical-related scientific or technical terms used herein shall have the meanings commonly understood by those skilled in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined solely by the claims.

[0156] All publications, patents, and patent applications are incorporated herein by reference in their entirety and are incorporated by reference to the same extent as if each individual publication, patent, and patent application was specifically and individually indicated to be incorporated by reference in its entirety. (Terms and Definitions)

[0157] The term "psilocybin" refers to the compound having the structure shown in Figure 1.

[0158] The term "indole prototype structure" refers to the chemical structure shown in Figure 2. Note that specific carbon and nitrogen atoms of the indole prototype structure are numbered. References may be made herein to such carbon and nitrogen numbers (e.g., C2, C4, N1, etc.). Furthermore, references may be made to chemicals of the indole prototype structure attached according to the same numbering, e.g., R4 and R6 reference chemicals attached to the C4 and C6 atoms, respectively. In addition, R 3A and R 3B refers to the reference chemical that extends from the ethylamino group that extends from the C3 atom of the indole prototype structure.

[0159] As used herein, the term "psilocybin derivative" refers to compounds derivable from psilocybin, including compounds having the indole prototype structure and the following formula (LXXVIII): [ka] and C3 ethylamine or ethylamine-derived groups having the formula 3a and R 3b are each independently a halogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3cis a halogen atom or a carboxy group). Psilocybin derivatives include compounds having one or more substituents at each of C2, C4, C5, C6, and C7. Thus, in formula (LXXVIII), R2, R4, R5, R6, and R7 can each be, for example, any one of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, (ix) a nitrile group, (x) an O-alkyl group, (xi), (xii) an O-acyl group, (xiii) a phosphate group, or (xiv) a halogen atom.

[0160] The term "polysubstituted psilocybin derivative" refers to a psilocybin derivative compound in which two or more substitution moieties are attached to psilocybin or a psilocybin derivative. The specific carbon atom substituted may be referenced. Furthermore, the substitution moieties may be referred to as S1, S2, S3, or S4, where S1, S2, S3, and S4 each represent a different substitution moiety. For example, a 5,7-S1,S2-di-psilocybin derivative refers to a psilocybin derivative in which carbon atoms 5 and 7 (as specified by the indole prototypical structure) each have a different substitution moiety. Similarly, a 2,5,6-tri-S1,S2,S3-psilocybin derivative refers to a psilocybin derivative in which carbon atoms 2, 5, and 6 (as specified by the indole prototypical structure) each have a different substitution moiety (or at least two of the three substitution moieties are different). As another example, a 2,5,6-tri-S1,S2,S2-psilocybin derivative refers to a psilocybin derivative in which carbon atoms 2, 5, and 6 (as defined by the indole prototypical structure) each have a substituted moiety, and the substituted moieties at carbon atoms 5 and 6 are identical. It should also be noted that, herein, S1, S2, S3, and S4 may additionally have subscript numbers, such as S15, S36, and S47. When such numbers are included, reference is made to the numbered C atoms of the prototypical structure. Thus, for example, S15 is a substituted moiety extending from the C5 atom of the indole ring structure, and S37 is a substituted moiety extending from the C7 atom of the indole ring structure. The term polysubstituted psilocybin derivative refers to a compound represented by the following formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group). The term polysubstituted psilocybin derivatives refers to compounds having the following formula (IV): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and each unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group). The term further includes salts of poly-substituted psilocybin, such as sodium and potassium salts.

[0161] As used herein, the terms "halogen," "halogenated," and "halo" refer to the class of elements consisting of fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). Halogenated compounds may therefore be referred to as "fluorinated," "chlorinated," "brominated," or "iodinated" compounds.

[0162] As used herein, the term "phosphate group" or "phospho group" refers to a molecule containing one phosphorus atom covalently bonded to four oxygen atoms (three single bonds and one double bond). One of the four oxygen atoms may be a hydroxy group, and one of the non-hydroxylated oxygen atoms may be chemically bonded to another moiety.

[0163] As used herein, the terms "hydroxy group" and "hydroxy" refer to a molecule containing an oxygen atom (having the formula -OH) that is bonded to a hydrogen atom. The hydroxy group may be chemically bonded to another moiety through the oxygen atom.

[0164] As used herein, "nitro group" or "nitro" refers to a molecule containing a nitrogen atom bonded to two oxygen atoms (having the formula -NO2). The nitro group may be chemically bonded to another moiety through the nitrogen atom. Additionally, a moiety bonded to a nitro group may be referred to herein as a "nitrated" moiety (e.g., a nitrated psilocybin derivative is a psilocybin derivative having a nitro group).

[0165] As used herein, "amino group" or "amino" refers to a molecule containing one nitrogen atom bonded to a hydrogen atom (having the formula -NH2). An amino group is protonated and has the formula -NH3 +The amino group may have the formula: The amino group may be chemically bonded to another moiety through its nitrogen atom. Additionally, a group bonded to an amino group may be referred to herein as an "aminated" moiety (e.g., an aminated psilocybin derivative is a psilocybin derivative having an amino group or an N-substituted amino group).

[0166] As used herein, the term "N-substituted amino group" refers to an amino group in which at least one of the hydrogen atoms has been replaced with another atom or group, such as an alkyl group, an acyl group, an aryl group, a sulfonyl group, or the like. The N-substituted amino group may be protonated, and the amino group may be chemically bonded to another moiety through its nitrogen atom. Thus, an N-substituted amino group, as used herein, is defined as follows: [ka] Furthermore, the N-substituted amino group includes the following bond: That is, chemical substance (IV) (alkyl group, aryl group): [ka] Chemical (V): [ka] Chemical Substance (VI) (Acyl Group): [ka] Chemical Substance (VII) (Sulfonyl Group): [ka] and chemical substance (VIII) (sulfonic acid group): [ka]

[0167] As used herein, "carboxyl group," "carboxyl," and "carboxy" refer to a molecule containing one carbon atom attached to an oxygen atom and a hydroxy group and having the formula -COOH. A carboxyl group includes a deprotonated carboxyl group, i.e., a group of the formula -COO - In the deprotonated form, the carboxyl group can be dissolved in a carboxyl salt, such as a sodium or potassium carboxyl salt, or an organic carboxyl salt (all herein referred to as COO - M + (represented by the formula:). It should be further understood that a carboxyl group may be chemically bonded to another moiety through its carbon atom. Furthermore, herein, a moiety bonded to a carboxyl group may be referred to as a "carboxylated" moiety (e.g., a carboxylated psilocybin derivative is a psilocybin derivative having a carboxyl group or an OH-substituted carboxyl group).

[0168] As used herein, the term "carboxylic acid derivative" refers to a carboxylic acid group in which the hydroxy group of the carboxylic acid group has been replaced by another atom or group, such as an -OR" group or an -NR'R" group. Thus, a carboxylic acid derivative includes the following chemical group (III): [ka]

[0169] As used herein, the term "aldehyde" or "aldehyde group" refers to a group of the formula: JPEG2024506667000093.jpg11159 (or may be further represented as -CHO). In this specification, -CHO may also be referred to as a formyl group. The aldehyde may be chemically bonded to another moiety through that carbon atom.

[0170] As used herein, the term "ketone" or "ketone group" refers to a molecule containing two carbon atoms, the first carbon atom being double-bonded to an oxygen atom, the first carbon atom being further bonded to a second carbon atom, and the molecule having the formula JPEG2024506667000094.jpg14159, where R is any moiety or moieties that allows the carbon atom attached to R to achieve normal valence. Thus, for example, R may represent three hydrogen atoms, or R may represent two hydrogen atoms and a methyl group. The ketone may be chemically bonded through its first carbon atom to another moiety (e.g., an alkylene group, (C1-C6) alkylene).

[0171] As used herein, a "nitrile group" or "nitrile" refers to a group attached to a nitrogen atom and having the formula JPEG2024506667000095.jpg9159 refers to a molecule containing one carbon atom having a nitrile group. It should be understood that the nitrile group may be chemically bonded to another moiety through that carbon atom. Additionally, herein, a moiety bonded to a nitrile group may be referred to as a "nitrilated" moiety (e.g., a nitrilated psilocybin derivative is a psilocybin derivative having a nitrile group).

[0172] As used herein, "glycosylated" or "glycosyl" refers to a saccharide group, such as a monosaccharide group, disaccharide group, trisaccharide group, oligosaccharide group, or polysaccharide group, in the form of a pyranose or furanose, in the α- or β-configuration, that can be attached or is attached from the anomeric carbon. When attached to another moiety through an oxygen atom by the anomeric carbon, the attached saccharide group, including the oxygen atom, is referred to herein as a "glycosyloxy" group. Exemplary monosaccharide groups include, but are not limited to, pentosyl, hexosyl, or heptosyl groups. Glycosyloxy groups may be substituted with various groups. Such substitutions may include lower alkyl, lower alkoxy, acyl, carboxy, carboxyamino, amino, acetamido, halo, thio, nitro, keto, and phosphatyl groups, where such substitutions may occur at one or more positions on the saccharide. The term glycosyl includes additional stereoisomers, optical isomers, anomers, and epimers of glycosyloxy groups. Thus, a hexose group may be, for example, an aldose or ketose group, may be in the D or L configuration, may be in the α or β configuration, and may be dextrorotatory or levorotatory relative to plane-polarized light. Exemplary glycosyloxy groups include, but are not limited to, glucosyl, galactosyl, fucosyl, xylose, arabinose, and rhamnose groups.

[0173] As used herein, a "prenyl group" or "prenyl" refers to the following structure (LVI): [ka] means a chemical group having the following structure: [ka] Further included are polyprenyl compounds having the formula: wherein n is an integer having a value of 2 or greater, eg, 2, 3, 4, 5, etc. Furthermore, the term "prenyl compound" refers to a compound that is, is substantially, or has a reactive prenyl group, i.e., a prenyl group that can be accepted by another moiety. Prenyl compounds include, for example, geranyl pyrophosphate (GPP), dimethylallyl diphosphate (DMAPP), farnesyl pyrophosphate (FPP), and geranylgeranyl pyrophosphate (GGPP).

[0174] As used herein, the term "alkyl group" refers to a group having 1 to "p" carbon atoms ("C1-C p "-alkyl" refers to straight and / or branched chain, saturated alkyl radicals containing "-alkyl" and, depending on the identity of "p," includes methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t-butyl, 2,2-dimethylbutyl, n-pentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, n-hexyl, etc., where p is an integer representing the maximum number of carbon atoms in the alkyl radical. Alkyl groups further include those of the formula -C n H 2n+1 Also included are linear hydrocarbon groups having the formula: including, but not limited to, methyl (-CH), ethyl (-C2H5), propyl (-C3H7), and butyl (-C4H9), as well as cyclic alkyl groups including cyclopropane, cyclobutane, cyclopentane, cyclohexane, and cycloheptane.

[0175] The term "cycloalkyl" means a cyclic alkyl group (C3-C 20 ), (C3-C 10 ), and (C3-C6) cycloalkyl groups, and further includes cyclopropane, cyclobutane, cyclopentane, cyclohexane, and cycloheptane.

[0176] As used herein, the term "O-alkyl group" refers to a group of the formula -OC n H 2n+1O-alkyl groups include, but are not limited to, O-methyl (-O-CH), O-ethyl (-O-C2H5), O-propyl (-O-C3H7), and O-butyl (-O-C4H9).

[0177] As used herein, the term "aryl group" means a hydrocarbon group arranged in an aromatic ring, e.g., C-C 14 -Aryl, C6-C 10 -aryl, etc. The aryl group further includes a phenyl group, a naphthyl group, a tetrahydronaphthyl group, a phenanthrenyl group, a biphenylenyl group, an indanyl group, a tolyl group, a xylyl group, an indenyl group, etc.

[0178] As used herein, the term "acyl group" refers to a carbon atom double-bonded to oxygen and single-bonded to an alkyl group. Such a carbon atom may be further bonded to another moiety. An acyl group has the chemical formula -C(=O)-C n H 2n+1 It can be described by

[0179] As used herein, the term "O-acyl group" refers to an acyl group in which a carbon atom is single-bonded to an additional oxygen atom. The additional oxygen atom may be bonded to another moiety. An O-acyl group is represented by the formula -OC(=O)-C n H 2n+1 Additionally, depending on the carbon chain, length-specific O-acyl groups may be referred to as acetoxy groups (n=1), propanoyloxy (n=2), butyryloxy (n=3), pentanoyloxy (n=4), etc.

[0180] As used herein, the term "alcohol group" or "hydroxyl alkyl" refers to a group having the chemical formula C n H n+1It refers to a hydrocarbon group arranged in a chain with OH. Depending on the carbon chain, the length-specific group may be called a methanol group (n=1) or hydroxymethyl, an ethanol group (n=2) or hydroxyethyl, a propanol group (n=3) or hydroxypropyl, a butanol group (n=4) or hydroxybutyl, etc.

[0181] As used herein, "5-HT 2A The term "5-HT receptor" refers to a subclass of the receptor family for the neurotransmitter and peripheral signal mediator serotonin. 2A The receptor can mediate multiple central and peripheral physiological functions of serotonin. Central nervous system effects include mediating the hallucinogenic effects of hallucinogenic compounds.

[0182] As used herein, "5-HT 2A The term "receptor modulation" refers to the 2A It refers to the ability of the compounds disclosed herein to alter the function of a 5-HT receptor. 2A Receptor modulators include 5-HT 2A It stimulates receptor activity and 5-HT 2A Depending on the concentration of the compound that contacts the receptor, 5-HT 2A It can stimulate or inhibit the activity of 5-HT receptors 2A Such activation or inhibition may occur upon the occurrence of a specific event, e.g., activation of a signal transduction pathway, and / or may be expressed only in specific cell types. 2A The term "modulation of 5-HT receptors" refers to 2A 5-HT by increasing or decreasing the probability that a complex will form between the receptor and its natural binding partner, promoting the formation of multimers. 2A It also means altering the function of the 5-HT receptor. 2A Receptor modulators include 5-HT 2A It may increase the probability of complex formation between the receptor and its natural binding partner, or it may increase the probability of 5-HT 2ADepending on the concentration of the compound that contacts the receptor, 5-HT 2A They may increase or decrease the probability of complex formation between the receptor and its natural binding partner, or they may increase or decrease the probability of 5-HT 2A In some cases, prenylated psilocybin derivatives may decrease the likelihood of complex formation between the receptor and its natural binding partner. 2A 5-HT by functioning as an agonist or antagonist of the receptor 2A and that the prenylated psilocybin derivatives of the present disclosure may alter the function of 5-HT 2A 5-HT receptors by interacting with or binding to them, or by interacting indirectly through one or more other molecular components. 2A Note that it may also alter receptor function.

[0183] As used herein, "5-HT 2A The term "receptor-mediated disorders" refers to 2A It is a disorder characterized by abnormal activity of 5-HT receptors. 2A Receptor-mediated disorders include 5-HT 2A It may be fully or partially regulated by receptor modulation, especially 5-HT 2A Receptor-mediated disorders include 5-HT 2A Receptor modulation may have some effect on the underlying disorder (e.g., 5-HT 2A administration of a receptor modulator results in some improvement in at least some of the treated subjects.

[0184] As used herein, "5-HT 1A The term "5-HT receptor" refers to a subclass of the receptor family for the neurotransmitter and peripheral signal mediator serotonin. 1A The receptors can mediate multiple central and peripheral physiological functions of serotonin. Many hallucinogenic compounds are 5-HT 1A Although known to modulate receptors and result in complex physiological responses, 5-HT1A Ligand activity of is not generally associated with hallucinogenesis (Inserra et al., 2020, Pharmacol Rev 73:202).

[0185] As used herein, "5-HT 1A The term "receptor modulation" refers to the 1A It refers to the ability of the compounds disclosed herein to alter the function of a 5-HT receptor. 1A Receptor modulators include 5-HT 1A It stimulates receptor activity and 5-HT 1A Depending on the concentration of the compound that contacts the receptor, 5-HT 1A It can stimulate or inhibit the activity of 5-HT receptors 1A Such activation or inhibition may occur upon the occurrence of a specific event, e.g., activation of a signal transduction pathway, and / or may be expressed only in specific cell types. 1A The term "modulation of 5-HT receptors" refers to 1A 5-HT by increasing or decreasing the probability that a complex will form between the receptor and its natural binding partner, promoting the formation of multimers. 1A It also means altering the function of the 5-HT receptor. 1A Receptor modulators include 5-HT 1A It may increase the probability of complex formation between the receptor and its natural binding partner, or it may increase the probability of 5-HT 1A Depending on the concentration of the compound that contacts the receptor, 5-HT 1A They may increase or decrease the probability of complex formation between the receptor and its natural binding partner, or they may increase or decrease the probability of 5-HT 1A In some cases, prenylated psilocybin derivatives may decrease the likelihood of complex formation between the receptor and its natural binding partner. 1A 5-HT by functioning as an agonist or antagonist of the receptor 1A and that the prenylated psilocybin derivatives of the present disclosure may alter the function of 5-HT 1A5-HT receptors by interacting with or binding to them, or by interacting indirectly through one or more other molecular components. 1A Note that it may also alter receptor function.

[0186] As used herein, "5-HT 1A The term "receptor-mediated disorders" refers to 1A It is a disorder characterized by abnormal activity of 5-HT receptors. 1A Receptor-mediated disorders include 5-HT 1A It may be fully or partially regulated by receptor modulation, especially 5-HT 1A Receptor-mediated disorders include 5-HT 1A Receptor modulation may have some effect on the underlying disorder (e.g., 5-HT 1A administration of a receptor modulator results in some improvement in at least some of the treated subjects.

[0187] As used herein, the term "reactive psilocybin derivative compound" refers to a psilocybin derivative compound that can react in a synthetic or biosynthetic reaction to form another psilocybin derivative compound, and generally includes an indole structure-containing reactant.

[0188] As used herein, the term "psilocybin derivative precursor compound" refers to a compound that serves as a precursor compound in the synthesis or biosynthesis of polysubstituted psilocybin derivatives, including compounds having the indole prototype structure, such as tryptophan and tryptamine, and further having the following formula (LVII): [ka] (wherein at least one of R2, R4, R5, R6, or R7 is a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and when unsubstituted R2, R5, R6, or R7 is each a hydrogen atom, and R4 is not substituted with any of the above substituents, then R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R3 is a hydrogen atom, -CH2-CHNH2COOH, or -CH2-CH2NH2.

[0189] As used herein, the term "psilocybin biosynthetic enzyme complement" refers to one or more polypeptides that, alone or in combination, facilitate the chemical conversion of psilocybin derivative precursor compounds to form polysubstituted psilocybin derivative compounds. The psilocybin biosynthetic enzyme complement includes, for example, one or more of tryptophan synthase B polypeptide, tryptophan decarboxylase, N-acetyltransferase, N-methyltransferase, and prenyltransferase.

[0190] As used herein, the term "tryptophan synthase B polypeptide" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence constituting any tryptophan synthase B polypeptide described herein (e.g., SEQ ID NO: 2), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any tryptophan synthase B polypeptide described herein.

[0191] As used herein, the term "tryptophan decarboxylase" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence making up any of the tryptophan decarboxylase polypeptides described herein (e.g., SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any of the tryptophan decarboxylases described herein.

[0192] As used herein, the term "N-acetyltransferase" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence making up any of the acetyltransferase polypeptides described herein (e.g., SEQ ID NO: 10), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any of the nucleic acid sequences encoding any of the acetyltransferases described herein.

[0193] As used herein, the term "N-methyltransferase" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence making up any of the N-methyltransferase polypeptides described herein (e.g., SEQ ID NO: 12 and SEQ ID NO: 14), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any of the N-methyltransferases described herein.

[0194] As used herein, the term "prenyltransferase" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence making up any of the prenyltransferase polypeptides described herein (e.g., SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, and SEQ ID NO:22), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any of the prenyltransferases described herein.

[0195] As used herein, the term "PsiH" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence constituting any PsiH polypeptide described herein (e.g., SEQ ID NO: 24), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any PsiH described herein.

[0196] As used herein, the term "CPR" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence constituting any CPR polypeptide described herein (e.g., SEQ ID NO: 26), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any CPR described herein.

[0197] As used herein, the term "PsiK" refers to any and all enzymes having (i) a sequence of amino acid residues substantially identical to the amino acid sequence constituting any PsiK polypeptide described herein (e.g., SEQ ID NO: 49), or (ii) a sequence of amino acid residues encoded by a nucleic acid sequence that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding any PsiK described herein.

[0198] The terms "nucleic acid sequence encoding a tryptophan synthase B polypeptide" and "nucleic acid sequence encoding a tryptophan synthase B polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences that encode a tryptophan synthase B polypeptide (e.g., SEQ ID NO: 1). A nucleic acid sequence encoding a tryptophan synthase B polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to a tryptophan synthase B polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize under at least moderately stringent hybridization conditions with any of the tryptophan synthase B polypeptide nucleic acid sequences described herein, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize under at least moderately stringent hybridization conditions.

[0199] The terms "nucleic acid sequence encoding a tryptophan decarboxylase" and "nucleic acid sequence encoding a tryptophan decarboxylase polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences that encode a tryptophan decarboxylase polypeptide (e.g., SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7). A nucleic acid sequence encoding a tryptophan decarboxylase polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to a tryptophan decarboxylase polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize under at least moderately stringent hybridization conditions with any of the tryptophan decarboxylase polypeptide nucleic acid sequences described herein, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize under at least moderately stringent hybridization conditions.

[0200] The terms "nucleic acid sequence encoding an N-acetyltransferase" and "nucleic acid sequence encoding an N-acetyltransferase polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences that encode an N-acetyltransferase polypeptide (e.g., SEQ ID NO: 9). A nucleic acid sequence encoding an N-acetyltransferase polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to an N-acetyltransferase polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize with any N-acetyltransferase polypeptide nucleic acid sequence described herein under at least moderately stringent hybridization conditions, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize with it under at least moderately stringent hybridization conditions.

[0201] The terms "nucleic acid sequence encoding an N-methyltransferase" and "nucleic acid sequence encoding an N-methyltransferase polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences that encode an N-methyltransferase polypeptide (e.g., SEQ ID NO: 11 and SEQ ID NO: 13). A nucleic acid sequence encoding an N-methyltransferase polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to an N-methyltransferase polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize with any N-methyltransferase polypeptide nucleic acid sequence described herein under at least moderately stringent hybridization conditions, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize with it under at least moderately stringent hybridization conditions.

[0202] The terms "nucleic acid sequence encoding a prenyltransferase" and "nucleic acid sequence encoding a prenyltransferase polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences that encode a prenyltransferase polypeptide (e.g., SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, and SEQ ID NO: 21). A nucleic acid sequence encoding a prenyltransferase polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to a prenyltransferase polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize with any of the prenyltransferase polypeptide nucleic acid sequences described herein under at least moderately stringent hybridization conditions, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize with it under at least moderately stringent conditions.

[0203] The terms "nucleic acid sequence encoding PsiH" and "nucleic acid sequence encoding a PsiH polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences encoding PsiH (e.g., SEQ ID NO: 23). A nucleic acid sequence encoding a PsiH polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to a PsiH polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize with any PsiH polypeptide nucleic acid sequence described herein under at least moderately stringent hybridization conditions, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize with it under at least moderately stringent hybridization conditions.

[0204] The terms "nucleic acid sequence encoding a CPR" and "nucleic acid sequence encoding a CPR polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences encoding a CPR (e.g., SEQ ID NO: 25). A nucleic acid sequence encoding a CPR polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to a CPR polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize with any CPR polypeptide nucleic acid sequence described herein under at least moderately stringent hybridization conditions, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize with it under at least moderately stringent hybridization conditions.

[0205] The terms "nucleic acid sequence encoding PsiK" and "nucleic acid sequence encoding a PsiK polypeptide" may be used synonymously herein and refer to any and all nucleic acid sequences encoding PsiK (e.g., SEQ ID NO: 48). A nucleic acid sequence encoding a PsiK polypeptide further includes (i) any and all nucleic acid sequences that encode a polypeptide substantially identical to a PsiK polypeptide sequence described herein, or (ii) any and all nucleic acid sequences that hybridize with any PsiK polypeptide nucleic acid sequence described herein under at least moderately stringent hybridization conditions, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize with it under at least moderately stringent hybridization conditions.

[0206] As used herein, the terms "nucleic acid" or "nucleic acid sequence" refer to a sequence of nucleoside or nucleotide monomers consisting of naturally occurring bases, sugars, and intersugar (backbone) linkages. The terms also include modified or substituted sequences containing non-naturally occurring monomers or portions thereof. Nucleic acids of the present disclosure may be deoxyribonucleic acids (DNA) or ribonucleic acids (RNA) and include naturally occurring bases, including adenine, guanine, cytosine, thymidine, and uracil. Examples of such modified bases include aza- and deaza-adenine, guanine, cytosine, thymidine, uracil, xanthine, and hypoxanthine. A sequence of nucleotide or nucleoside monomers may be referred to as a polynucleotide sequence, nucleic acid sequence, nucleotide sequence, or nucleoside sequence.

[0207] The term "polypeptide," as used herein in connection with a reference SEQ ID NO, means any and all polypeptides containing (i) a sequence of amino acid residues that is substantially identical to the amino acid sequence making up the polypeptide having such reference SEQ ID NO, or (ii) a sequence of amino acid residues that, in the absence of synonymous codon usage, is capable of hybridizing under at least moderately stringent conditions to any nucleic acid sequence encoding a polypeptide having such reference SEQ ID NO. A sequence of amino acid residues may be referred to as an amino acid sequence or a polypeptide sequence.

[0208] The term "nucleic acid sequence encoding a polypeptide," as used herein in reference to a reference SEQ ID NO, refers to any and all nucleic acid sequences that encode the polypeptide having such reference SEQ ID NO. A nucleic acid sequence encoding a polypeptide in reference to a reference number further includes any and all nucleic acid sequences that encode a polypeptide substantially identical to the polypeptide having such reference SEQ ID NO, or (ii) any and all nucleic acid sequences that hybridize with the nucleic acid sequence encoding the polypeptide having such reference SEQ ID NO under at least moderately stringent hybridization conditions, or, in the absence of synonymous codon usage, any and all nucleic acid sequences that hybridize with it under at least moderately stringent conditions.

[0209] The term "substantially identical" means that two amino acid sequences are preferably at least 70% identical, more preferably at least 85% identical, and most preferably at least 95% identical (e.g., 96%, 97%, 98%, or 99% identical). To determine the percent identity between two amino acid sequences, the two amino acid sequences are aligned using, for example, the alignment method of Needleman and Wunsch (J. Mol. Biol. 1970, 48:443), as revised by Smith and Waterman (Adv. Appl. Math. 1981, 2:482), so that the best match is obtained between the two sequences and the number of identical amino acids between the two sequences is determined. Methods for calculating percent identity between two amino acid sequences are generally recognized techniques, and include, for example, the method described by Carillo and Lipton (SIAM J. Applied Math., 1988, 48:1073) and the method described by Lesk, in Computational Molecular Biology, ed., Oxford University Press, New York, 1988, Biocomputing: Informatics and Genomics Projects. Such calculations generally utilize computer programs. Computer programs used in such methods include, but are not limited to, GCG (Devereux et al., Nucleic Acid Pres., 1984, 12:387), BLASTP, BLASTN, and FASTA (Altschul et al., J. Mol. Biol., 1990, 215:403).Particularly preferred methods for determining percent identity between two polypeptides include the Clustal W algorithm (Thompson, JD, Higgines, DG and Gibson TJ, 1994, Nucleic Acid Pres 22(22):4673-4680), in conjunction with the BLOSUM 62 scoring matrix (Henikoff S & Henikoff, JG, 1992, Proc. Natl. Acad. Sci. USA 89: 10915-10919), using a gap opening penalty of 10 and a gap extension penalty of 0.1, so as to align at least 50% of the entire length of one of the two sequences to obtain the best match between the two sequences.

[0210] The term "at least moderately stringent hybridization conditions" refers to conditions selected to promote selective hybridization between two complementary nucleic acid molecules in solution. Hybridization may occur across the entire nucleic acid sequence molecule or across a portion thereof. The hybridized portion is typically at least 15 (e.g., 20, 25, 30, 40, or 50) nucleotides in length. Those skilled in the art will recognize that the stability of a nucleic acid duplex or hybrid is determined by its Tm, which is a function of sodium ion concentration and temperature in a sodium-containing buffer (Tm = 81.5°C - 16.6 (Log10[Na+]) + 0.41 (%(G+C) - 600 / L or similar equation). Thus, the washing condition parameters that determine hybrid stability are sodium ion concentration and temperature. To identify molecules that are similar but not identical to a known nucleic acid molecule, a 1% mismatch may be assumed to result in a decrease in Tm of approximately 1°C. For example, if a nucleic acid molecule with greater than 95% identity is desired, In this case, the final wash temperature will be reduced by about 5°C. Those skilled in the art will be able to easily select appropriate hybridization conditions based on such considerations. In a preferred embodiment, stringent hybridization conditions are selected. For example, to achieve stringent hybridization conditions, the following conditions may be used: hybridization with 5x sodium chloride / sodium citrate (SSC) / 5x Denhardt's solution / 1.0% SDS at Tm-5°C (based on the above equation), followed by washing with 0.2x SSC / 0.1% SDS at 60°C. Moderately stringent hybridization conditions include a wash step with 3x SSC at 42°C. However, equivalent stringency can be achieved with alternative buffers, salts, and It should be understood that this can also be achieved using temperature. Additional guidance regarding hybridization conditions may be found in Current Protocols in Molecular Biology, John Wiley & Sons, NY, 1989, 6.3.1-6.3.6 and Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory Press, 1989, Vol. 3.

[0211] The term "functional variant," as used herein in reference to a polynucleotide or polypeptide, refers to a polynucleotide or polypeptide capable of performing the same function as the reference polynucleotide or polypeptide of interest. Thus, for example, a functional variant of the polypeptide set forth in SEQ ID NO: 2 refers to a polypeptide capable of performing the same function as the polypeptide set forth in SEQ ID NO: 2. Functional variants also include modified polypeptides, where such modifications include the substitution, deletion, or addition of one or more amino acids. In some embodiments, a substitution replaces an amino acid with an amino acid of similar characteristics. Such substitutions include, but are not limited to, (i) glutamic acid and aspartic acid, (ii) alanine, serine, and threonine, (iii) isoleucine, leucine, and valine, (iv) asparagine and glutamine, and (v) tryptophan, tyrosine, and phenylalanine. Functional variants further include polypeptides that retain or exhibit improved psilocybin biosynthetic biological activity.

[0212] The term "chimera," as used herein in reference to nucleic acids, refers to two linked nucleic acids that are not naturally linked. Chimeric nucleic acids include linked nucleic acids from different natural sources. For example, a nucleic acid comprising a microbial promoter linked to a nucleic acid encoding a plant polypeptide is considered a chimera. Chimeric nucleic acids also include nucleic acids from the same natural source that are not naturally linked. For example, a nucleic acid comprising a promoter from a particular cell type is linked to a nucleic acid encoding a polypeptide from the same cell type that is not normally linked to the nucleic acid comprising the promoter. Chimeric nucleic acids also include nucleic acids having any naturally occurring nucleic acid linked to any nucleic acid that does not naturally occur.

[0213] As used herein, the term "pharmaceutical formulation" means a preparation of an active ingredient (including psychoactive ingredients) in a form that allows it to provide effective therapy, but without any other ingredients that would cause excessive toxicity, allergic reaction, early inflammatory reaction, or other adverse reaction that is commensurate with a reasonable risk / benefit ratio. A pharmaceutical formulation may also include other pharmaceutical ingredients, such as excipients, carriers, diluents, or adjuvants.

[0214] As used herein, the term "recreational drug formulation" refers to a preparation containing psychoactive ingredients in a form that allows for effective recreational drug administration, but that does not contain any other ingredients that would cause excessive toxicity, allergic reaction, early inflammatory reaction, or other adverse effects that are commensurate with a reasonable risk / benefit ratio. Recreational drug formulations may also include other ingredients, such as excipients, carriers, diluents, or adjuvants.

[0215] As used herein, the term "effective for administration as a recreational drug" refers to a formulation (typically self-administered) that is administered non-medically by a subject at will to induce psychoactive effects, which may include altered states of consciousness, gratification, pleasure, ecstasy, altered perceptions, or hallucinations.

[0216] As used herein, the term "effective amount" refers to an amount of an active ingredient, pharmaceutical formulation, or recreational drug formulation sufficient to induce a desired biological or therapeutic effect, including preventative and psychoactive effects. Such effects include effects on the signs, symptoms, or causes of a disorder or disease, or any other desired alteration of a biological system. Effective amounts vary depending, for example, on the health status, stage of injury, stage of disease, or disorder, weight or sex of the subject being treated, timing of administration, method of administration, and age of the subject, all of which can be determined by one of ordinary skill in the art.

[0217] As used herein, the terms "treat," "treatment," and the like mean to obtain a desired physiological, pharmacological, or biological effect, including prophylactic or therapeutic treatment. Such an effect may result in the suppression, alleviation, amelioration, or reversal of signs, symptoms, or causes of a disorder or disease attributable to such disorder or disease. Clinical evidence of prevention or treatment may vary depending on the disease or disorder, the subject, and the treatment selected.

[0218] As used herein, the term "pharmaceutically acceptable" means a substance, including excipients, carriers, diluents, or adjuvants, that are compatible with the substance in a pharmaceutical or recreational drug formulation and that, within the scope of sound medical judgment, can be used in contact with a subject without excessive toxicity, allergic response, early inflammatory response, or other adverse reaction commensurate with a reasonable risk / benefit ratio.

[0219] The terms "substantially pure" and "isolated," when used interchangeably herein, refer to a compound (e.g., a psilocybin derivative) that has been separated from components that naturally coexist with it. Generally, a compound is substantially pure when at least 60%, more preferably at least 75%, more preferably at least 90%, 95%, 96%, 97%, or 98%, and most preferably at least 99% of the total material in a sample (based on volume, wet or dry weight, mole percent, or mole fraction) is the compound of interest. Purity can be measured by any appropriate method, such as chromatography, gel electrophoresis, or HPLC analysis.

[0220] The term "recovered," as used herein in connection with an enzyme, protein, or compound, means a substantially pure form of such enzyme, protein, or compound.

[0221] The term "in vivo," as used herein in connection with methods for making poly-substituted psilocybin derivative compounds, refers to a method comprising the step of converting a psilocybin derivative precursor compound to a poly-substituted psilocybin derivative compound by contacting the psilocybin derivative precursor compound with an enzyme capable of converting the psilocybin derivative precursor compound, e.g., in cells (e.g., cells or microorganisms) cultured in a growth medium. Such cells typically express a psilocybin biosynthetic enzyme complex, including heterologously expressed tryptophan synthase B polypeptide, tryptophan decarboxylase, N-acetyltransferase, N-methyltransferase, and prenyltransferase.

[0222] The term "in vitro," as used herein in connection with methods for making poly-substituted psilocybin derivative compounds, refers to a method comprising the step of contacting a psilocybin derivative precursor compound outside a cell, e.g., in a microwell plate, tube, flask, beaker, tank, reactor, etc., with an enzyme capable of converting such psilocybin derivative precursor compound to a poly-substituted psilocybin derivative compound. (General Practice)

[0223] As mentioned above, the present disclosure relates to psilocybin derivatives. In particular, the present disclosure provides novel polysubstituted psilocybin derivatives. The compositions provided herein generally exhibit functional properties that deviate from those of psilocybin. Thus, for example, polysubstituted psilocybin derivatives may exhibit pharmacological properties that deviate from those of psilocybin. Furthermore, polysubstituted psilocybin derivatives may exhibit physicochemical properties different from those of psilocybin. Thus, for example, polysubstituted psilocybin derivatives may exhibit excellent solubility in solvents (e.g., aqueous solvents). In this respect, polysubstituted psilocybin derivatives are useful in the formulation of pharmaceutical and recreational drug formulations. In one embodiment, the polysubstituted psilocybin derivatives of the present disclosure can be easily produced chemically and / or biosynthetically. Such a method avoids the process of extracting psilocybin from mushrooms and then performing a chemical reaction to obtain the polysubstituted derivative. Furthermore, the growth of mushrooms is avoided, thus limiting dependence on weather and climate and limiting potential legal and social challenges associated with cultivating mushrooms containing psychoactive compounds. Such methods allow for the efficient production of vast quantities of polysubstituted psilocybin derivatives.

[0224] In the following, selected embodiments are described with reference to the drawings.

[0225] First, exemplary poly-substituted psilocybin derivatives are described, followed by methods of using and making the poly-substituted psilocybin derivatives.

[0226] Thus, in one aspect, the present disclosure provides derivatives of the compound known as psilocybin, the chemical structure of which is shown in Figure 1. The derivatives provided herein are, in particular, multi-substituted derivatives, including psilocybin derivatives having two or more substituted moieties.

[0227] Thus, in at least one embodiment of one aspect, the present disclosure provides, in accordance with the teachings herein, a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group).

[0228] Thus, referring to a compound of formula (I), it is first noted that in one embodiment thereof, at least two of R2, R4, R5, R6, or R7 are substitution moieties selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group. Thus, it should be understood that in one embodiment of the present disclosure, at least two of R2, R4, R5, R6, or R7 are substitution moieties. The replacement moieties are each independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, wherein if two (but not more than two) replacement moieties are selected, they are non-identical, and if more than two replacement moieties are selected, at least two of the selected replacement moieties are non-identical.

[0229] In one embodiment of one aspect of the present specification, at least three of R2, R4, R5, R6, or R7 may be a substituent selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0230] In one embodiment of one aspect of the present specification, at least three of R2, R4, R5, R6, or R7 may be a substituent selected from at least three of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0231] Continuing with reference to compounds of formula (I), in a further embodiment herein, R4 may be a phosphate group or a hydrogen atom.

[0232] Continuing with reference to compounds of formula (I), in a further embodiment herein, R 3a and R 3b may be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group. 3a and R 3b may be a hydrogen atom, or R 3a and R 3b may each be an alkyl group, e.g., methyl, ethyl, propyl, or a longer chain alkyl group, or R 3a and R 3b may each be an acyl group, or R 3a and R 3b may each be an aryl group. 3a and R 3b One of the atoms is a hydrogen atom, and R 3a and R3b One of R may be an alkyl group. 3a and R 3b One of the atoms is a hydrogen atom, and R 3a and R 3b One of R may be an acyl group. 3a and R 3b One of the atoms is a hydrogen atom, and R 3a and R 3b One of R may be an aryl group. 3a and R 3b One of the groups is an alkyl group, and R 3a and R 3b One of R may be an aryl group. 3a and R 3b One of the groups is an alkyl group, and R 3a and R 3b One of R may be an acyl group. 3a and R 3b One of the groups is an acyl group, and R 3a and R 3b One of the groups may be an aryl group.

[0233] Continuing with reference to compounds of formula (I), in a further embodiment herein, each unsubstituted R, R, R, or R group is a hydrogen atom. Additionally, as noted above, R may also be a hydrogen atom.

[0234] In accordance with the present disclosure, in one embodiment, polysubstituted psilocybin derivatives have two or three substituents. Each of these examples is discussed below with reference to selected figures. In particular, examples of polysubstituted psilocybin derivatives having two substituents are discussed with reference to Figures 3A-3L, and examples of polysubstituted psilocybin derivatives having three substituents are discussed with reference to Figures 4A-4I, 5A-5I, 6A-6I, and 7A-7I. For clarity, note that in each of these figures, designations such as S15, S25, and S17 refer to a first substituent S1 at the 5-position (C5), a second substituent S2 at the 5-position (C5), and a first substituent S1 at the 7-position (C7), respectively. Similarly, within a particular figure, designations such as S22 and S26 within the same compound refer to a second substituent S2 at the 2-position (C2) and the same second substituent at the 6-position (C6) of the same compound (see, e.g., Figure 6I).

[0235] Thus, referring now to Figures 3A-3L (examples of multiply substituted psilocybin derivatives according to the present invention in which two of R2, R4, R5, R6, or R7 are substituents), a 2,5-di-S1,S2-psilocybin derivative is shown in Figure 3A, a 2,5-di-S2,S1-psilocybin derivative is shown in Figure 3B, a 2,6-di-S1,S2-psilocybin derivative is shown in Figure 3C, a 2,6-di-S2,S1-psilocybin derivative is shown in Figure 3D, and a 2,7-di-S1,S2-psilocybin derivative is shown in Figure 3E. 2,7-di-S2,S1-psilocybin derivatives are shown in FIG. 3F, 5,6-di-S1,S2-psilocybin derivatives are shown in FIG. 3G, 5,6-di-S2,S1-psilocybin derivatives are shown in FIG. 3H, 5,7-di-S1,S2-psilocybin derivatives are shown in FIG. 3I, 5,7-di-S2,S1-psilocybin derivatives are shown in FIG. 3J, 6,7-di-S1,S2-psilocybin derivatives are shown in FIG. 3K, and 6,7-di-S2,S1-psilocybin derivatives are shown in FIG. 3L. As is apparent from the foregoing, in each of Figures 3A to 3L, S1 and S2 are selected from two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0236] Thus, for example, with reference to a compound of formula (I), in one exemplary embodiment, two of R4, R5, R6, or R7 may be substituents, one of which may be selected from (i) a halogen atom, (ii) a prenyl group, and (iii) a nitrile group, and one of R4, R5, R6, or R7 may be a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, provided that the two substituents are non-identical and R2 and the unsubstituted R4, R5, R6, or R7 are hydrogen atoms. 3A-3L, in such exemplary embodiments, S1 may be selected from (i) a halogen atom, (ii) a prenyl group, and (iii) a nitrile group, and S2 may be selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; or conversely, S2 may be selected from (i) a halogen atom, (ii) a prenyl group, and (iii) a nitrile group, and S1 may be selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0237] Thus, for example, referring to a compound of formula (I), in one further exemplary embodiment, two of R4, R5, R6, or R7 may be substituents, one of which is a halogen atom, and one of R4, R5, R6, or R7 may be a substituent selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, and (viii) a nitrile group, and R2 and unsubstituted R4, R5, R6, or R7 are hydrogen atoms. 3A-3L, in such exemplary embodiments, S1 may be a halogen atom and S2 may be selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, and (viii) a nitrile group; or conversely, S2 may be a halogen atom and S1 may be (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, and (viii) a nitrile group.

[0238] Thus, for example, with reference to compounds of Formula (I), in one further exemplary embodiment, two of R, R, R, or R may be substituents, one of which is a halogen atom, and one of R, R, R, or R may be a substituent selected from (i) an amino group, (ii) a nitrile group, (iii) a nitro group, (iv) a hydroxy group, and (vii) a prenyl group, and R and the unsubstituted R, R, R, or R are hydrogen atoms. Thus, with reference to Figures 3A-3L, in such exemplary embodiments, S may be a halogen atom and S may be selected from (i) an amino group, (ii) a nitrile group, (iii) a nitro group, (iv) a hydroxy group, and (vii) a prenyl group, or conversely, S may be a halogen atom and S may be (i) an amino group, (ii) a nitrile group, (iii) a nitro group, (iv) a hydroxy group, and (v) a prenyl group.

[0239] Thus, for example, with reference to compounds of Formula (I), in one further exemplary embodiment, two of R, R, R, or R may be substituents, one of which is a halogen atom, and one of R, R, R, or R may be a prenyl group, and R and the unsubstituted R, R, R, or R are hydrogen atoms. Thus, with reference to Figures 3A-3L, in such exemplary embodiments, S may be a halogen atom and S may be a prenyl group, or conversely, S may be a prenyl group and S may be a halogen atom.

[0240] Thus, for example, referring to a compound of formula (I), in one further exemplary embodiment, two of R4, R5, R6, or R7 may be substituents, one of which is a prenyl group, and one of R4, R5, R6, or R7 may be a substituent selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a halogen atom, and (viii) a nitrile group, and R2 and unsubstituted R4, R5, R6, or R7 are hydrogen atoms. 3A-3L, in such exemplary embodiments, S1 may be a prenyl group and S2 may be selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a halogen atom, and (viii) a nitrile group; or conversely, S2 may be a prenyl group and S1 may be (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a halogen atom, and (viii) a nitrile group.

[0241] Thus, for example, with reference to compounds of Formula (I), in one further exemplary embodiment, two of R4, R5, R6, or R7 may be substituents, one of which is a prenyl group, and one of R4, R5, R6, or R7 may be a substituent selected from (i) a carboxyl group or a carboxylic acid derivative, (ii) a halogen atom, and (iii) a hydroxy group, and R2 and the unsubstituted R4, R5, R6, or R7 are hydrogen atoms. Thus, with reference to Figures 3A-3L, in such exemplary embodiments, S1 may be a prenyl group and S2 may be selected from (i) a carboxyl group or a carboxylic acid derivative, (ii) a halogen atom, and (iii) a hydroxy group, or conversely, S2 may be a prenyl group and S1 may be (i) a carboxyl group or a carboxylic acid derivative, (ii) a halogen atom, and (iii) a hydroxy group.

[0242] Thus, for example, referring to a compound of formula (I), in one further exemplary embodiment, two of R4, R5, R6, or R7 may be substituents, one of which is a nitrile group, and one of R4, R5, R6, or R7 may be a substituent selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, or (viii) a halogen atom, and R2 and unsubstituted R4, R5, R6, or R7 are hydrogen atoms. 3A-3L, in such exemplary embodiments, S1 may be a nitrile group and S2 may be selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, or (viii) a halogen atom; or conversely, S2 may be a nitrile group and S1 may be (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, or (viii) a halogen atom.

[0243] Thus, for example, with reference to compounds of Formula (I), in one further exemplary embodiment, two of R4, R5, R6, or R7 may be substituents, one of which is a nitrile group, and one of R4, R5, R6, or R7 may be an amino group or an N-substituted amino group, and R2 and the unsubstituted R4, R5, R6, or R7 are hydrogen atoms. Thus, with reference to Figures 3A-3L, in such exemplary embodiments, S1 may be a nitrile group and S2 may be an amino group or an N-substituted amino group, or conversely, S2 may be a nitrile group and S1 may be an amino group or an N-substituted amino group.

[0244] Next, referring to Figures 4A-4F, 4G-4I, 5A-5F, 5G-5I, 6A-6F, 6G-6I, 7A-7F, and 7G-7I, examples of polysubstituted psilocybin derivatives according to the present specification are shown, in which three of R2, R5, R6, or R7 are substituents. Note that in the examples shown in Figures 4A-4F, 5A-5F, 6A-6F, 6G-6I, and 7A-7F, the three substituents are three different substituents, S1, S2, and S3. In Figures 4G-4I, 5G-5I, 6G-6I, and 7G-7I, the three substituents are selected so that two of the three substituents are identical (i.e., S1, S2, and S2). In this regard, for example, among others, 2,5,6-tri-S1,S2,S3-psilocybin derivatives are shown in Figure 4A, 2,5,6-tri-S1,S3,S2-psilocybin derivatives are shown in Figure 4B, 2,5,6-tri-S3,S1,S2-psilocybin derivatives are shown in Figure 4C, 2,5,6-tri-S2,S1,S3-psilocybin derivatives are shown in Figure 4D, and 2,5,6-tri-S2,S3,S1-psilocybin derivatives are shown in Figure 4F. As is clear from the above description, in each of Figures 4A to 4F, S1, S2, and S3 are selected from three of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0245] Further examples in which three of R2, R5, R6, or R7 are substituents are shown in Figures 4G, 4H, and 4I, particularly a 2,5,6-tri-S1,S2,S2-psilocybin derivative shown in Figure 4G, a 2,5,6-tri-S2,S1,S2-psilocybin derivative shown in Figure 4H, and a 2,5,6-tri-S2,S2,S1-psilocybin derivative shown in Figure 4I. As is clear from the foregoing, in each of Figures 4G-4I, S1 and S2 are selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0246] Further examples in which three of R2, R5, R6, or R7 are substituents are shown in Figures 5A, 5B, 5C, 5D, 5E, and 5F, and in particular, a 2,5,7-tri-S1,S2,S3-psilocybin derivative is shown in Figure 5A, a 2,5,7-tri-S1,S3,S2-psilocybin derivative is shown in Figure 5B, a 2,5,7-tri-S3,S1,S2-psilocybin derivative is shown in Figure 5C, a 2,5,7-tri-S2,S1,S3-psilocybin derivative is shown in Figure 5D, a 2,5,7-tri-S3,S2,S1-psilocybin derivative is shown in Figure 5E, and a 2,5,7-tri-S2,S3,S1-psilocybin derivative is shown in Figure 5F. As is clear from the foregoing description, in each of Figures 5A to 5I, S1, S2, and S3 are selected from three of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0247] Further examples in which three of R2, R5, R6, or R7 are substituents are shown in Figures 5G, 5H, and 5I, particularly a 2,5,7-tri-S1,S2,S2-psilocybin derivative shown in Figure 5G, a 2,5,7-tri-S2,S1,S2-psilocybin derivative shown in Figure 5H, and a 2,5,7-tri-S2,S2,S1-psilocybin derivative shown in Figure 5I. As is clear from the foregoing, in each of Figures 5G-5I, S1 and S2 are selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0248] Further examples in which three of R2, R5, R6, or R7 are substituents are shown in Figures 6A, 6B, 6C, 6D, 6E, and 6F, and in particular, a 2,6,7-tri-S1,S2,S3-psilocybin derivative is shown in Figure 6A, a 2,6,7-tri-S1,S3,S2-psilocybin derivative is shown in Figure 6B, a 2,6,7-tri-S3,S1,S2-psilocybin derivative is shown in Figure 6C, a 2,6,7-tri-S2,S1,S3-psilocybin derivative is shown in Figure 6D, a 2,6,7-tri-S3,S2,S1-psilocybin derivative is shown in Figure 6E, and a 2,6,7-tri-S2,S3,S1-psilocybin derivative is shown in Figure 6F. As is clear from the above description, in each of Figures 6A to 6F, S1, S2, and S3 are selected from three of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0249] Further examples in which three of R2, R5, R6, or R7 are substituents are shown in Figures 6G, 6H, and 6I, particularly a 2,6,7-tri-S1,S2,S2-psilocybin derivative shown in Figure 6G, a 2,6,7-tri-S2,S1,S2-psilocybin derivative shown in Figure 6H, and a 2,6,7-tri-S2,S2,S1-psilocybin derivative shown in Figure 6I. As is clear from the foregoing, in each of Figures 6A-6I, S1 and S2 are selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0250] Further examples in which three of R2, R5, R6, or R7 are substituents are shown in Figures 7A, 7B, 7C, 7D, 7E, and 7F, and in particular, a 5,6,7-tri-S1,S2,S3-psilocybin derivative is shown in Figure 7A, a 5,6,7-tri-S1,S3,S2-psilocybin derivative is shown in Figure 7B, a 5,6,7-tri-S3,S1,S2-psilocybin derivative is shown in Figure 7C, a 5,6,7-tri-S2,S1,S3-psilocybin derivative is shown in Figure 7D, a 5,6,7-tri-S3,S2,S1-psilocybin derivative is shown in Figure 7E, and a 5,6,7-tri-S2,S3,S1-psilocybin derivative is shown in Figure 7F. As is clear from the above description, in each of Figures 7A to 7F, S1, S2, and S3 are selected from three of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0251] Further examples in which three of R2, R5, R6, or R7 are substituents are shown in Figures 7G, ​​7H, and 7I, particularly a 5,6,7-tri-S1,S2,S2-psilocybin derivative shown in Figure 7G, a 5,6,7-tri-S2,S1,S2-psilocybin derivative shown in Figure 7H, and a 5,6,7-tri-S2,S2,S1-psilocybin derivative shown in Figure 7I. As is clear from the foregoing, in each of Figures 7G-7I, S1 and S2 are selected from two of: (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0252] Furthermore, in the exemplary embodiments shown in 3A-3L, 4A-4I, 5A-5I, 6A-6I, and 7A-7I, R 3a and R 3b Note that R may be a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group. 3a and R 3b may be a hydrogen atom, or R 3a and R 3b may each be an alkyl group, e.g., methyl, ethyl, propyl, or a longer chain alkyl group, or R 3a and R 3b may each be an acyl group, or R 3a and R 3b may each be an aryl group. 3a and R 3b One of the atoms is a hydrogen atom, and R 3a and R 3b One of R may be an alkyl group. 3a and R 3b One of the atoms is a hydrogen atom, and R 3a and R 3b One of R may be an acyl group. 3a and R 3b One of the atoms is a hydrogen atom, and R3a and R 3b One of R may be an aryl group. 3a and R 3b One of the groups is an alkyl group, and R 3a and R 3b One of R may be an aryl group. 3a and R 3b One of the groups is an alkyl group, and R 3a and R 3b One of R may be an acyl group. 3a and R 3b One of the groups is an acyl group, and R 3a and R 3b One of the groups may be an aryl group.

[0253] In each of the exemplary embodiments set forth in 3A-3L, 4A-4I, 5A-5I, 6A-6I, and 7A-7I in further aspects of the present specification, R 3c Note that may be a halogen atom or a carboxy group.

[0254] In further aspects of the present specification, in each of the exemplary embodiments shown in 3A-3L, 4A-4I, 5A-5I, 6A-6I, and 7A-7I, it is noted that R4 can be a halogen atom, an O-alkyl group, an O-acyl group, or a phosphate group.

[0255] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (IX): [ka] It may also be a compound of the formula:

[0256] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (X): [ka] It may also be a compound of the formula:

[0257] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XI): [ka] It may also be a compound of the formula:

[0258] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XII): [ka] It may also be a compound of the formula:

[0259] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XIII): [ka] It may also be a compound of the formula:

[0260] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XIV): [ka] It may also be a compound of the formula:

[0261] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XV): [ka] It may also be a compound of the formula:

[0262] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XV): [ka] It may also be a compound of the formula:

[0263] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XVII): [ka] It may also be a compound of the formula:

[0264] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XVIII): [ka] It may also be a compound of the formula:

[0265] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XIX): [ka] It may also be a compound of the formula:

[0266] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XX): [ka] It may also be a compound of the formula:

[0267] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXI): [ka] It may also be a compound of the formula:

[0268] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXII): [ka] It may also be a compound of the formula:

[0269] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXIII): [ka] It may also be a compound of the formula:

[0270] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXIV): [ka] It may also be a compound of the formula:

[0271] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXV): [ka] It may also be a compound of the formula:

[0272] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XVI): [ka] It may also be a compound of the formula:

[0273] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXVII): [ka] It may also be a compound of the formula:

[0274] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXVIII): [ka] It may also be a compound of the formula:

[0275] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXIX): [ka] It may also be a compound of the formula:

[0276] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXX): [ka] It may also be a compound of the formula:

[0277] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXI): [ka] It may also be a compound of the formula:

[0278] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXII): [ka] It may also be a compound of the formula:

[0279] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXIII): [ka] It may also be a compound of the formula:

[0280] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXIV): [ka] It may also be a compound of the formula:

[0281] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXV): [ka] It may also be a compound of the formula:

[0282] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXVI): [ka] It may also be a compound of the formula:

[0283] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXVII): [ka] It may also be a compound of the formula:

[0284] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXVIII): [ka] It may also be a compound of the formula:

[0285] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XXXIX): [ka] It may also be a compound of the formula:

[0286] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XL): [ka] It may also be a compound of the formula:

[0287] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLI): [ka] It may also be a compound of the formula:

[0288] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLII): [ka] It may also be a compound of the formula:

[0289] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLIII): [ka] It may also be a compound of the formula:

[0290] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLIV): [ka] It may also be a compound of the formula:

[0291] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLV): [ka] It may also be a compound of the formula:

[0292] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLVI): [ka] It may also be a compound of the formula:

[0293] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLVII): [ka] It may also be a compound of the formula:

[0294] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLVIII): [ka] It may also be a compound of the formula:

[0295] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (XLIX): [ka] It may also be a compound of the formula:

[0296] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (L): [ka] It may also be a compound of the formula:

[0297] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (LI): [ka] It may also be a compound of the formula:

[0298] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (LII): [ka] It may also be a compound of the formula:

[0299] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (LIII): [ka] It may also be a compound of the formula:

[0300] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (LIV): [ka] It may also be a compound of the formula:

[0301] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (LV): [ka] It may also be a compound of the formula:

[0302] Furthermore, in one exemplary embodiment, the poly-substituted psilocybin derivatives of the present disclosure have the following formula (LXXVI): [ka] It may also be a compound of the formula:

[0303] It should also be noted that the polysubstituted psilocybin derivatives of the present disclosure include salts thereof, including pharmaceutically acceptable salts. Thus, the nitrogen atom of the ethylamino group extending from the C3 atom may be protonated and its positive charge may be balanced with, for example, a chloride ion or a sulfate ion to form a chloride salt or a sulfate salt. Furthermore, in compounds where R4 is a phosphate group, the phosphate group may be deprotonated and its negative charge may be balanced with, for example, a sodium ion or a potassium ion to form a sodium salt or a potassium salt.

[0304] Furthermore, when R4 is a phosphate group, the term prenylated psilocybin derivative refers to a compound represented by the following formula (IV): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and each unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c Note that R is a hydrogen atom or a carboxyl group. 3c is a carboxy group, the following formula (Iva): [ka] Further included are salts (e.g., sodium salts, potassium salts, etc.) of the prenylated psilocybin derivatives of formula (IV) and (IVa).

[0305] Thus, briefly summarized, the present disclosure provides polysubstituted psilocybin derivatives, particularly those having the following formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group).

[0306] (C1-C 20 ) In another embodiment, R 3a and R 3b is a hydrogen atom, (C1-C 20 ) alkyl group, (C6-C 14 ) aryl group, or -C(=O)(C1-C 20 ) alkyl group. In another embodiment, R 3a and R 3b is a hydrogen atom, (C1-C 10 ) alkyl group, (C6-C 10 ) aryl group, or -C(=O)(C1-C 10 ) alkyl group. In another embodiment, R 3a and R 3b is a hydrogen atom, a (C1-C6) alkyl group, a phenyl group, or a -C(=O)(C1-C6) alkyl group. 3a and R 3b is a hydrogen atom, a methyl group, an ethyl group, a propyl group, a phenyl group, -C(=O)-CH3, -C(=O)-CH2CH3, or -C(=O)-CH2CH2CH3.

[0307] In another embodiment, R 3a and / or R 3b is (C1-C 20 ) cycloalkyl group, or (C1-C 10 ) cycloalkyl group, (C1-C 10 ) cycloalkyl group, or (C1-C 10 ) cycloalkyl group. In one embodiment, R 3a and / or R 3b is a cyclopropane group, a cyclobutane group, a cyclopentane group, or a cyclohexane group.

[0308] In one embodiment, the alkyl group (including O-alkyl groups) in any one of the formula definitions of the present disclosure is C-C 20 In another embodiment, the alkyl group is C-C 10 In another embodiment, the alkyl group is C1-C6 alkyl. In another embodiment, the alkyl group is methyl, ethyl, propyl, butyl, or pentyl.

[0309] In one embodiment, the acyl group (including O-acyl group) in any one of the formula definitions of the present disclosure is C1-C 20 Acyl (or C1-C 20 In another embodiment, the alkyl group is C-C 10 Acyl (or C1-C 10 In another embodiment, the alkyl group is C1-C6 acyl (or C1-C6 acyl-O). In another embodiment, the acyl group is an O-acyl group, methanoyl, ethanoyl, propanoyl, butanoyl, or pentanoyl.

[0310] In one embodiment, the aryl group in any one of the formula definitions of the present disclosure is an optionally substituted C-C 14 In another embodiment, the aryl group is an optionally substituted C-C 10 In another embodiment, the aryl group is phenyl, naphthyl, tetrahydronaphthyl, phenanthrenyl, biphenylenyl, indanyl, indenyl, and the like.

[0311] The poly-substituted psilocybin derivatives of the present disclosure may be used in the preparation of pharmaceutical or recreational drug formulations. Accordingly, in one embodiment of another aspect, the present disclosure further provides pharmaceutical or recreational drug formulations having poly-substituted psilocybin derivatives. Accordingly, in a further embodiment of one aspect, the present disclosure provides a compound having the following formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group).

[0312] Pharmaceutical or recreational drug formulations may be formulated as liquids, tablets, capsules, microcapsules, nanocapsules, transdermal patches, gels, foams, aerosols, nanoparticles, powders, creams, emulsions, micellar systems, films, sprays, ovules, infusions, teas, decoctions, suppositories, and the like, including pharmaceutically acceptable salts or solvates of nitrated psilocybin derivatives containing excipients. The term "excipient" refers to any ingredient other than the compounds of the present disclosure. As will be readily apparent to those skilled in the art, the selection of an excipient may be based on factors such as the particular method of administration and the effect of the excipient on the solubility of the compounds of the present disclosure, and preparation methods will be apparent to those skilled in the art. Such compositions and formulation methods may be found, for example, in "Remington's Pharmaceutical Sciences," 22nd Edition (Pharmaceutical Press and Philadelphia College of Pharmacy at the University of the Sciences, 2012).

[0313] Dosages for use of the compounds of the present disclosure can vary within wide limits and can be tailored to the individual circumstances of each individual case, as is customary and known to those skilled in the art. The dosage will depend, for example, on the nature and severity of the disease being treated, the condition of the patient, the compound being applied, whether acute or chronic treatment is being performed, whether prophylaxis is being performed, the method of compound delivery, and whether additional active compounds are administered in addition to the compounds of the present disclosure. Representative dosages of the present disclosure include, but are not limited to, about 0.001 mg to about 5000 mg, about 0.001 mg to about 2500 mg, about 0.001 mg to about 1000 mg, about 0.001 mg to about 500 mg, about 0.001 mg to about 250 mg, about 0.001 mg to about 100 mg, about 0.001 mg to about 50 mg, and about 0.001 mg to about 25 mg. Representative dosages of the present disclosure include, but are not limited to, about 0.0001 to about 1,000 mg, about 10 to about 160 mg, about 10 mg, about 20 mg, about 40 mg, about 80 mg, or about 160 mg. Multiple dosages may be administered daily, e.g., 2, 3, or 4 dosages, particularly when relatively large amounts are deemed necessary. It may be necessary to vary the dosages set forth herein upward or downward, depending on the subject's condition and as deemed appropriate by the patient's physician or caregiver.

[0314] Pharmaceutical or recreational drug formulations containing the disclosed polysubstituted psilocybin derivatives may be administered orally. Oral administration may involve swallowing, which allows the compound to enter the gastrointestinal tract, or buccal or sublingual administration, which allows the compound to enter the bloodstream directly from the mouth. Formulations suitable for oral administration include both solid and liquid formulations.

[0315] Solid formulations include tablets, capsules (containing particles, liquids, microcapsules, or powders), lozenges (including liquid-filled lozenges), chewables, multiparticulates or nanoparticles, gels, solid solutions, liposomal formulations, microencapsulated formulations, creams, films, ovules, suppositories, and sprays.

[0316] Liquid formulations include suspensions, solutions, syrups and elixirs.Such formulations can be used as fillers for soft or hard capsules, and generally contain carriers such as water, ethanol, polyethylene glycol, propylene glycol, methylcellulose or suitable oils, and one or more emulsifiers and / or suspending agents.Liquid formulations can also be prepared by reconstituting solids, for example, from sachets.

[0317] Binders are commonly used to impart cohesive properties to tablets. Suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose, and hydroxypropyl methylcellulose.

[0318] Tablets may also contain diluents, such as lactose (monohydrate, spray-dried monohydrate, anhydrous, etc.), mannitol, xylitol, glucose, sucrose, sorbitol, microcrystalline cellulose, starch, and dibasic calcium phosphate dihydrate.

[0319] Tablets may optionally have a surfactant such as sodium lauryl sulfate, polysorbate 80, etc. If a surfactant is used, it may comprise 0.2% (w / w) to 5% (w / w) of the tablet.

[0320] Tablets may further comprise a lubricant such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl furamate, or a mixture of magnesium stearate and sodium lauryl sulfate. Lubricants generally comprise from 0.25% (w / w) to 10% (w / w), or from 0.5% (w / w) to 3% (w / w) of the tablet.

[0321] In addition to the poly-substituted psilocybin derivative, the tablet may contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropylcellulose, starch, pregelatinized starch, and sodium alginate. Disintegrants typically comprise 1% (w / w) to 25% (w / w), or 5% (w / w) to 20% (w / w) of the dosage form.

[0322] Other possible adjuvants include antioxidants, colorants, fragrances, preservatives, and flavor enhancers.

[0323] With respect to tablet dosage forms, the compounds of the present disclosure may comprise from 1% (w / w) to 80% (w / w) of the dosage form, more typically from 5% (w / w) to 60% (w / w) of the dosage form, depending on the desired effective amount of the compound.

[0324] An exemplary tablet has about 80% (w / w) or less of the compound, about 10% (w / w) to about 90% (w / w) of a binder, about 0% (w / w) to about 85% (w / w) of a diluent, about 2% (w / w) to about 10% (w / w) of a disintegrant, and about 0.25% (w / w) to about 10% (w / w) of a lubricant.

[0325] Tablet formation is discussed in "Pharmaceutical Dosage Forms: Tablets," Vol. 1-Vol. 3, by CPC Press (2008).

[0326] Pharmaceutical or recreational drug formulations containing the disclosed polysubstituted psilocybin derivatives may be administered directly into the bloodstream, muscle, or internal organs. Thus, pharmaceutical or recreational drug formulations may be administered parenterally (e.g., by subcutaneous, intravenous, intraarterial, intrathecal, intraventricular, intracerebral, intramuscular, or intraperitoneal injection). Parenteral formulations are generally aqueous solutions (in one embodiment, pH 3 to 9) containing excipients (salts, carbohydrates, buffers, etc.), but may more suitably be formulated as sterile non-aqueous solutions or as a dry form for use with suitable excipients, such as sterile water, depending on the application.

[0327] Formulations containing the disclosed polysubstituted psilocybin derivatives for parenteral administration may be formulated for immediate and / or modified release. Modified release formulations include delayed-release, sustained-release, pulsed-release, controlled-release, targeted-release, and programmed-release. Thus, the disclosed compounds may be formulated as solids, semisolids, or thixotropic liquids for administration as implanted depots that provide modified release of the active compound. Examples of such formulations include drug-coated stents and poly(dl-lactic-coglycolic) acid (PGLA) microspheres.

[0328] The pharmaceutical or recreational drug formulations of the present disclosure may be administered topically to the skin or mucosa, i.e., transdermally. Exemplary pharmaceutical or recreational drug formulations for such purposes include gels, hydrogels, lotions, liquids, creams, ointments, dusting powders, cosmetics, oils, eye drops, wound dressings, foams, films, skin patches, wafers, implants, sponges, fibers, bandages, and microemulsions. Liposomes may also be used. Exemplary carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. Penetration enhancers may also be used (see, e.g., Finnin, B. and Morgan, TM, 1999 J. Pharm. Sci., 88(10), 955-958).

[0329] Other methods of topical administration include delivery by electroporation, iontophoresis, phonophoresis, sonophoresis, and microneedle or needle-free (eg, Powderject™, Bioject™, etc.) injection.

[0330] Pharmaceutical or recreational drug formulations for inhalation or aeration include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid pharmaceutical compositions may contain suitable pharmaceutically acceptable excipients. In some embodiments, pharmaceutical compositions are administered by the oral or nasal respiratory route for local or systemic effect. Pharmaceutical compositions in pharmaceutically acceptable solvents may be nebulized using an inert gas. Nebulized solutions may be inhaled directly from a nebulizing device, or the nebulizing device may be attached to a face mask tent or intermittent positive pressure breathing machine. Solution, suspension, or powder pharmaceutical compositions may be administered orally or nasally, for example, from a device that delivers the formulation in an appropriate manner.

[0331] In further embodiments in which the polysubstituted psilocybin derivatives of the present application are used as recreational drugs, the compounds may be included in compositions such as foods or food products, beverages, condiments, personal care products, e.g., cosmetics, perfumes, bath oils, or oils (such as topical application as massage oils or oils for burning or aerosolization).

[0332] It is therefore apparent that poly-substituted psilocybin derivative compounds may be used as pharmaceutical or recreational drugs. Accordingly, in at least one embodiment of another aspect herein, the present disclosure provides a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group).

[0333] Pharmaceutical formulations having compounds of the present disclosure may be used to treat a subject, particularly to treat a psychiatric disorder in a subject. Thus, in a further embodiment, the present invention includes a method for treating a psychiatric disorder, the method comprising administering to a subject a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3cis a hydrogen atom or a carboxyl group) to a subject in need thereof.

[0334] Psychiatric disorders treated include, for example, neurodevelopmental disorders such as intellectual disability, global developmental delay, communication disorders, autism spectrum disorders, and attention-deficit hyperactivity disorder (ADHD); bipolar disorders and related disorders such as manic depression; anxiety disorders such as generalized anxiety disorder (GAD); stress disorders such as agoraphobia, social anxiety disorder, specific phobias (of natural phenomena, medical conditions, animals, situations, etc.), panic disorder, separation anxiety disorder, acute stress disorder, adjustment disorder, post-traumatic stress disorder (PTSD), and reactive attachment disorder; dissociative disorders such as dissociative amnesia, dissociative identity disorder, and depersonalization / derealization syndrome; somatic symptom disorder, illness anxiety disorder, and conversion disorder. somatoform disorders such as sexual disorders and factitious disorder; eating disorders such as anorexia nervosa, anorexia nervosa, rumination disorder, pica, and binge eating disorder; sleep disorders such as narcolepsy, insomnia disorder, hypersomnia, breathing-related sleep disorder, parasomnia, and restless legs syndrome; disruptive behavior disorders such as kleptomania, pyromania, intermittent explosive disorder, post-traumatic stress disorder, and oppositional defiant disorder; depressive disorders caused by another medical condition (e.g., psychosocial and existential distress within life-threatening cancer symptoms) (ACS) Substance-related disorders include alcohol-related disorders, cannabis-related disorders, inhalant-related disorders, stimulant use disorders, and tobacco use disorders; neurocognitive disorders such as delirium; schizophrenia; obsessive-compulsive disorders such as obsessive-compulsive disorder (OCD), body dysmorphic disorder, hoarding disorder, trichotillomania, substance- or medication-induced obsessive-compulsive disorder, and obsessive-compulsive disorder associated with another medical condition; and personality disorders such as antisocial personality disorder, avoidant personality disorder, borderline personality disorder, dependent personality disorder, histrionic personality disorder, narcissistic personality disorder, obsessive-compulsive personality disorder, paranoid personality disorder, schizotypal personality disorder, and schizotypal personality disorder.

[0335] In one embodiment, the compounds of the present disclosure are 5-HT 2A , thereby contacting 5-HT2A Such contact may involve modulation of receptors, e.g., 5-HT 2A Receptor-containing sample, purified 5-HT 2A Samples containing receptors, or 5-HT 2A The compounds of the present disclosure and 5-HT are combined by introducing the compounds into a sample containing cells having the receptor. 2A The contacting step includes combining a compound of the invention with a 5-HT receptor under in vitro conditions. The in vitro conditions further include those described in Example 1 herein. The contacting step includes combining a compound of the invention with a 5-HT receptor under in vivo conditions. 2A Such in vivo conditions include administering a pharmaceutically effective amount of the compound to an animal or human subject when the compound is formulated with a pharmaceutically active carrier, diluent, or excipient, as described above, and treating the subject. 2A By contacting the receptor, 5-HT 2A Receptor activation or 5-HT 2A It blocks receptors.

[0336] Thus, in a further embodiment, the condition treated in accordance with the present invention is any 5-HT 2A The disorder may be a receptor-mediated disorder, including, but not limited to, schizophrenia, psychotic disorders, attention deficit hyperactivity disorder, autism, and bipolar disorder.

[0337] In one embodiment, the compounds of the present disclosure are 5-HT 1A , thereby contacting 5-HT 1A Such contact may involve modulation of receptors, e.g., 5-HT 1A Receptor-containing sample, purified 5-HT 1A Samples containing receptors, or 5-HT 1A The compounds of the present disclosure and 5-HT are combined by introducing the compounds into a sample containing cells having the receptor. 1A The contacting step includes combining a compound of the invention with a 5-HT receptor under in vitro conditions. The in vitro conditions further include those described in Example 1 herein. The contacting step includes combining a compound of the invention with a 5-HT receptor under in vivo conditions.1A Such in vivo conditions include administering a pharmaceutically effective amount of the compound to an animal or human subject when the compound is formulated with a pharmaceutically active carrier, diluent, or excipient, as described above, and treating the subject. 1A By contacting the receptor, 5-HT 1A Receptor activation or 5-HT 1A It blocks receptors.

[0338] Thus, in a further embodiment, the condition treated in accordance with the present invention is any 5-HT 1A The disorder may be a receptor-mediated disorder, including, but not limited to, schizophrenia, psychotic disorders, attention deficit hyperactivity disorder, autism, and bipolar disorder.

[0339] In some embodiments, the compound is 5-HT 1A Receptors and 5-HT 2A When exposed to the receptor, 5-HT 1A Modulates receptors (e.g., 5-HT 1A receptors), but at the same time 2A It may not regulate receptors.

[0340] In some embodiments, the compound is 5-HT 2A Receptors and 5-HT 1A When exposed to the receptor, 5-HT 2A Modulates receptors (e.g., 5-HT 2A receptors), but at the same time 1A It may not regulate receptors.

[0341] With regard to methods for making the polysubstituted psilocybin derivatives of the present invention, it should first be noted that the polysubstituted psilocybin derivatives of the present invention may be prepared by any suitable method, including any organic chemical synthetic method, biosynthetic method, or a combination thereof. Next, exemplary methods for chemically making the polysubstituted psilocybin derivatives of the present disclosure will first be discussed. Subsequently, exemplary biosynthetic methods for making the polysubstituted psilocybin derivatives will be discussed.

[0342] One suitable method for making the disclosed polysubstituted psilocybin derivatives includes selecting and obtaining or preparing a reactive psilocybin derivative compound and selecting and obtaining or preparing a substituent-containing compound, followed by chemically or biochemically reacting the reactive psilocybin derivative with the substituent-containing compound to obtain the polysubstituted psilocybin derivative compound. In embodiments herein in which the reactive psilocybin derivative compound does not already possess at least one substituent, such a reactive, unsubstituted psilocybin derivative compound (i.e., an indole structure-containing reactant in which R2, R5, R6, and R7 are each hydrogen atoms) can generally be reacted sequentially with at least two substituent-containing compounds. Examples are shown in Figures 9, 10, 11A-11C, and 12A-12D (showing exemplary reactions to form a first psilocybin derivative having a single substituent) and 13A-13C (showing exemplary subsequent reactions to form polysubstituted psilocybin derivatives having two or more substituents). In other embodiments, the reactive psilocybin derivative compound (as shown in Figures 8A-8G) has at least one substituent and is reacted with at least one additional substituent-containing compound.

[0343] Thus, in at least one embodiment of one aspect, the present disclosure provides a compound of formula (I): : [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group) or a salt thereof, the method comprising: [ka] wherein one of R2, R4, R5, R6, or R7 is a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom or an alcohol group; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, a hydroxy group, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3cis a hydrogen atom or a carboxyl group) with a substituent-containing compound, wherein the substituent of the substituent-containing compound is selected from (i) halogen-containing compounds, (ii) hydroxy-containing compounds, (iii) nitro-containing compounds, (iv) glycosyloxy-containing compounds, (v) amino- or N-substituted amino-containing compounds, (vi) carboxyl- or carboxylic acid derivative-containing compounds, (vii) aldehyde- or ketone-containing compounds, (viii) prenyl-containing compounds, and (ix) nitrile-containing compounds, under reaction conditions sufficient to form a psilocybin substituent of formula (I) or a salt thereof.

[0344] The reactive psilocybin derivative compound of formula (II) includes multiple compounds. Generally, the reactive psilocybin derivative compound of formula (II) can be selected by first identifying the desired multi-substituted psilocybin derivative compound, determining its substituents, and then selecting an appropriate reactive psilocybin derivative compound having formula (II). Thus, for example, if an S14,S26 multi-substituted psilocybin derivative is desired to be prepared, an S14 reactive psilocybin derivative compound can be selected and reacted with a compound containing an S2 substituent to form the desired S14,S26 multi-substituted psilocybin derivative; alternatively, if an S15,S26 multi-substituted psilocybin derivative is desired to be prepared, an S15 reactive psilocybin derivative compound can be selected and reacted with a compound containing an S2 substituent to form the desired S15,S26 multi-substituted psilocybin derivative. Thus, it can be more generally stated that carrying out chemical reactions to make compounds of the present disclosure involves substitution at different atoms (i.e., R2, R4, R5, R6, and / or R7 atoms).

[0345] Thus, in one exemplary embodiment, S15 is a chlorine atom and R2, R6, and R7 are S22, S2 6, To form a polysubstituted psilocybin derivative, such as S27, R4 is an O-alkyl group, R2, R6, and R7 are hydrogen atoms, R5 is a chlorine atom, and R 3a and R 3bThe reactive psilocybin derivative may be selected to result in a compound in which is a hydrogen atom, an alkyl group, an acyl group, or an aryl group.

[0346] In a further exemplary embodiment, S15 is a chlorine atom, and R2, R6, and R7 are S22, S2 6, To form a polysubstituted psilocybin derivative, such as S27, R4 is an O-acyl group, R2, R6, and R7 are hydrogen atoms, R5 is a chlorine atom, and R 3a and R 3b The reactive psilocybin derivative may be selected to result in a compound in which is a hydrogen atom, an alkyl group, an acyl group, or an aryl group.

[0347] In a further exemplary embodiment, S15 is a chlorine atom, and R2, R6, and R7 are S22, S2 6, To form a polysubstituted psilocybin derivative, such as S27, R4 is a hydroxyl group, R2, R6, and R7 are hydrogen atoms, R5 is a chlorine atom, and R 3a and R 3b The reactive psilocybin derivative may be selected to result in a compound in which is a hydrogen atom, an alkyl group, an acyl group, or an aryl group.

[0348] In a further exemplary embodiment, S15 is a chlorine atom, and R2, R6, and R7 are S22, S2 6, To form a polysubstituted psilocybin derivative, such as S27, R4 is a phosphate group, R2, R6, and R7 are hydrogen atoms, R5 is a chlorine atom, and R 3a and R 3b The reactive psilocybin derivative may be selected to result in a compound in which is a hydrogen atom, an alkyl group, an acyl group, or an aryl group.

[0349] In a further exemplary embodiment, S15 is a chlorine atom, and R2, R6, and R7 are S22, S2 6,To form a polysubstituted psilocybin derivative, such as S27, R4 is a hydrogen atom, R2, R6, and R7 are hydrogen atoms, R5 is a chlorine atom, and R 3a and R 3b The reactive psilocybin derivative may be selected to result in a compound in which is a hydrogen atom, an alkyl group, an acyl group, or an aryl group.

[0350] The reactive psilocybin derivative compounds may be provided in a substantially chemically pure form, e.g., in the form of a psilocybin derivative preparation having a purity of at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.9%. Psilocybin derivatives may be chemically synthesized or obtained from fine chemical manufacturers, e.g., Sigma-Aldrich® (St. Louis, MO, USA).

[0351] The substituent-containing compound may be any compound capable of reacting with a selected reactive psilocybin derivative compound and having a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group.

[0352] The substituent-containing compounds may be provided in a substantially chemically pure form, e.g., at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.9% pure. The nitrile-containing compounds may be chemically synthesized or purified, or may be obtained from fine chemical manufacturers, e.g., Sigma-Aldrich® (St. Louis, MO, USA).

[0353] For example, the examples shown in Figures 9, 10, 11A-11C, 12A-12D, and 13A-13C are exemplary reactions for initially forming nitrated (Figure 9), glycosylated (Figure 10), aminated (Figures 11A-11C), and carboxylated (Figures 12A-12C) psilocybin derivatives. The resulting compounds may then be reacted with additional substituent-containing compounds, as shown in the examples in Figures 13A-13C, respectively.

[0354] Therefore, referring to Figure 13A, this shows an exemplary chemical transformation for converting the initially synthesized 7-nitrated psilocybin derivative (see compound 13A-2) to other 5,7-disubstituted psilocybin derivatives containing two types of groups at the C5 and C7 carbon atoms. For example, Friedel-Crafts acylation of compound 13A-2 regioselectively adds an acyl group to the C5 carbon, providing compound 13A-3, which can be further oxidized to the corresponding 5-carboxy derivative (compound 13A-4) via an iodoform reaction. The 7-nitro group can then be reduced to the corresponding 7-amino psilocybin derivative (compound 13A-5) via palladium-mediated reduction using formic acid as the hydrogen source. The 7-amino group can then be further converted to a reactive and versatile diazonium salt (compound 13A-6) using sodium nitrite as a reagent under acidic conditions. The reactive 7-diazonium salt intermediate compound 13A-6 may give the corresponding 5-carboxypsilocybin derivative further containing 7-nitrile (compound 13A-7), 7-hydroxy (compound 13A-8), 7-fluorine (compound 13A-9), or 7-iodine (compound 13A-10) groups.

[0355] In one exemplary embodiment of this aspect, the first psilocybin derivative formed may be a compound having an acetyl group (e.g., the compound shown in Figure 13A, designation 13A-3). In other examples, R2, R4, or R6, or R7 may have an acetyl group.

[0356] Therefore, with further reference to Figure 13A and the compound of Formula (I), it will be apparent that the first poly-substituted psilocybin derivative compound formed may further react to form additional poly-substituted psilocybin derivative compounds. Thus, for example, the first poly-substituted psilocybin derivative formed may be the compound shown in Figure 13A, Designation 13A-3, or a compound in which other combinations of R2, R4, R5, R6, and R7 have nitro and acetyl groups. The first psilocybin derivative formed may react to oxidize the acetyl group to form a second psilocybin derivative of Formula (I) (wherein one or at least one of R2, R4, R5, R6, or R7 is a nitro group and one or at least one of R2, R4, R5, R6, or R7 is a carboxyl group), forming a second psilocybin derivative, for example, the compound shown in Figure 13A, Designation 13A-4.

[0357] With further reference to Figure 13A and the compound of Formula (I), the second psilocybin derivative formed may react to reduce the nitro group to form an amino group and a third psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an amino group and one or at least one of R2, R4, R5, R6, or R7 is a carboxyl group). The third psilocybin derivative formed may be the compound shown in Figure 13A, designation 13A-5.

[0358] With further reference to Figure 13A and the compound of Formula (I), the formed tertiary psilocybin derivative may be reacted with a nitrite salt to convert the amino group of the diazonium salt to form an intermediate psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is a diazonium group and one or at least one of R2, R4, R5, R6, or R7 is a carboxyl group). The formed intermediate psilocybin derivative may be, for example, the compound shown in Figure 13A, designation 13A-6.

[0359] With further reference to Figure 13A and the compound of Formula (I), the intermediate psilocybin derivative formed may be reacted with a nitrile-containing compound to convert the diazonium group and form a quaternary psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is a nitrile group and one or at least one of R2, R4, R5, R6, or R7 is a carboxyl group), such as the compound shown in Figure 13A, designation 13A-7.

[0360] With further reference to Figure 13A and the compound of Formula (I), the intermediate psilocybin derivative formed may react with water to convert the diazonium group and form a fifth psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is a hydroxy group and one or at least one of R2, R4, R5, R6, or R7 is a carboxyl group), such as the compound shown in Figure 13A, designation 13A-8.

[0361] With further reference to Figure 13A and the compound of Formula (I), the intermediate psilocybin derivative formed may be reacted with a halogen-containing compound to convert the diazonium group and form a sixth psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is a halogen atom and one or at least one of R2, R4, R5, R6, or R7 is a carboxyl group), such as the compounds shown in Figure 13A, Designations 13A-9 and 13A-10.

[0362] Referring now to Figure 13B, an exemplary chemical transformation is shown here, using the initially synthesized 5-carboxy psilocybin derivative (see compound Figure 13B-1) to convert it to other O-4-glycosylated psilocybin derivatives containing two types of groups at the C5 and C7 carbon atoms. Thus, starting with compound 13B-1, the 5-carboxy group may be selectively esterified in methanol under Fischer esterification conditions to provide compound 13B-2, which may then be used to synthesize the corresponding 7-chloro (compound 13B-3) and 7-nitro (compound 13B-4) derivatives, respectively. Both compounds 13B-3 and 13B-4 may be glycosylated at the O-4 position using per-O-acetylated glycosyl bromide to give the corresponding 4-O-glycosylated derivatives, compounds 13A-5 and 13A-6, after removal of all O-acetates. The 5-ester group in both compounds can be further converted to a 5-amide group via aminolysis to provide the 4-O-glycosylated psilocybin derivatives 13B-7 and 13B-8, respectively. The 7-nitro group in 13B-8 can be further reduced to provide compound 13B-9 via palladium-mediated reduction using formic acid as the hydrogen source.

[0363] With further reference to Figure 13B and the compound of Formula (I), it will be apparent that a reactive psilocybin derivative compound having a substituent may be reacted with a substituent-containing compound to form an initial poly-substituted psilocybin derivative compound, and that this first compound may be used to form additional poly-substituted psilocybin derivative compounds. Thus, for example, the substituent of the reactive psilocybin derivative may be a methoxycarbonyl group and the substituent-containing compound may be the halogen-containing compound N-halosuccinimide, and the reactive psilocybin derivative and the substituent-containing compound may react to form a first psilocybin derivative, e.g., the compound shown in Figure 13B, Designation 13B-3 (wherein one of R2, R4, R5, R6, or R7 is a methoxycarbonyl group and one or at least one of R2, R4, R5, R6, or R7 is a halogen atom).

[0364] With further reference to Figure 13B and the compounds of formulas (I) and (II), the reactive psilocybin derivative may have a methoxycarbonyl group, and the substituent-containing compound may be the nitro-containing compound nitronium tetrafluoroborate, and the reactive psilocybin derivative and the substituent-containing compound may react to form a second psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is a methoxycarbonyl group and one or at least one of R2, R4, R5, R6, or R7 is a nitro group), and the second psilocybin derivative formed may be the compound shown in 13B, designation 13B-4.

[0365] With further reference to Figure 13B and the compound of Formula (I), the first psilocybin derivative formed may be reacted with an acetylated glycosyl compound to form a third psilocybin (wherein one or at least one of R2, R4, R5, R6, or R7 is a methoxycarbonyl group, one or at least one of R2, R4, R5, R6, or R7 is a glycosyloxy group, and one or at least one of R2, R4, R5, R6, or R7 is a halogen atom), e.g., the compound shown in Figure 13B, designation 13B-5.

[0366] With further reference to Figure 13B and the compound of Formula (I), the second psilocybin derivative formed may be reacted with an acetylated glycosyl compound to form a fourth psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is a methoxycarbonyl group, one or at least one of R2, R4, R5, R6, or R7 is a glycosyloxy group, and one or at least one of R2, R4, R5, R6, or R7 is a nitro group). For example, the fourth psilocybin derivative formed may be the compound shown in Figure 13B, designation 13B-6.

[0367] With further reference to Figure 13B and the compound of Formula (I), the formed third psilocybin derivative may react with ammonia to convert the methoxycarbonyl group in the amide group to form a fifth psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an amide group, one or at least one of R2, R4, R5, R6, or R7 is a glycosyloxy group, and one or at least one of R2, R4, R5, R6, or R7 is a halogen atom), e.g., the compound shown in Figure 13B, designation 13B-7.

[0368] With further reference to Figure 13B and the compound of Formula (I), the formed third psilocybin derivative may react with ammonia to convert the methoxycarbonyl group in the amide group to form a sixth psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an amide group, one or at least one of R2, R4, R5, R6, or R7 is a glycosyloxy group, and one or at least one of R2, R4, R5, R6, or R7 is a nitro group), e.g., the compound shown in Figure 13B, designation 13B-8.

[0369] With further reference to Figure 13B and the compound of formula (I), the sixth psilocybin derivative formed may react to reduce the nitro group to form an amino group and a seventh psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an amide group, one or at least one of R2, R4, R5, R6, or R7 is a glycosyloxy group, and one or at least one of R2, R4, R5, R6, or R7 is a nitro group), e.g., the compound shown in Figure 13B, designation 13B-9.

[0370] Thus, referring to Figure 13C, this shows further exemplary chemical transformations for converting the synthesized 5-nitrated psilocybin derivative (see compound 13C-2) to 5,6,7-tri-substituted psilocybin derivatives containing up to three types of substituents. For example, the 5-nitro group may be reduced through palladium-mediated reduction in the presence of formic acid as a hydrogen source to produce the 5-amino derivative (compound 13C-3). Chemoselective N-acetylation in methanol may be carried out to provide the corresponding 5-acetamino psilocybin derivative 13C-4, which may then be formylated at carbon atom C7 using DMF-POCl3 as a reagent to provide the 7-aldehyde compound 13C-5. Further bromination using N-bromosuccinimide may provide the 5-acetamido-6-bromo-7-formyl psilocybin derivative 13C-6. Alternatively, compound 13C-5 can be used as a substituent and silver carbonate can be used as a reagent to oxidize the 7-formyl group under mild conditions to provide compound 13C-7, which can then be esterified using Fisher esterification conditions to provide compound 13C-8. 13C-8 can be reacted with nitrosonium tetrafluoroborate to introduce a nitro group at carbon atom C6 to provide compound 13C-9, which can then be reduced to form compound 13C-10, which can then be converted to an amide via aminolysis to provide novel 5,6,7-trisubstituted psilocybin derivative compound 13C-11.

[0371] With further reference to Figure 13C and the compounds of Formulae (I) and (II), it will be apparent that a reactive psilocybin derivative compound having a substituent may be reacted with a substituent-containing compound to form an initial poly-substituted psilocybin derivative compound, and that such a first compound may be used to form additional poly-substituted psilocybin derivative compounds. Thus, for example, the substituent of the reactive psilocybin derivative may be an acetamidyl group and the substituent-containing compound may be a halogen-containing compound, N-halosuccinimide (e.g., N-bromosuccinimide (NBS)), and the reactive psilocybin derivative and the substituent-containing compound may react to form a first psilocybin derivative, e.g., the compound shown in Figure 13C, Designation 13C-6 (wherein one or at least one of R2, R4, R5, R6, or R7 is an acetamidyl group and one or at least one of R2, R4, R5, R6, or R7 is a halogen atom).

[0372] With further reference to FIG. 13C and the compounds of Formulae (I) and (II), the reactive psilocybin derivative may have an acetamidyl group, the substituent-containing compound may be dimethylformamide, and the reactive psilocybin derivative and the substituent-containing compound may react to form an intermediate psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an acetamidyl group and one or at least one of R2, R4, R5, R6, or R7 is a methanol group), which may react to oxidize the methanol group to form a second psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an acetamidyl group and one or at least one of R2, R4, R5, R6, or R7 is a carboxy group). For example, the intermediate psilocybin derivative may be the compound shown in 13C, designation 13C-5, and the second psilocybin derivative formed may be the compound shown in 13C, designation 13C-7.

[0373] With further reference to Figure 13C and the compound of Formula (I), the formed second psilocybin derivative of Formula (I) may be reacted with an alcohol to esterify the carboxy group to form an ester and a third psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an acetamidyl group and one or at least one of R2, R4, R5, R6, or R7 is a carboxyl ester), such as the compound shown in Figure 13C, designation 13C-8.

[0374] With further reference to Figure 13C and the compound of Formula (I), the formed third psilocybin derivative may be reacted with a nitro-group-containing compound to form a fourth psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an acetamidyl group, one or at least one of R2, R4, R5, R6, or R7 is a carboxyl ester, and one or at least one of R2, R4, R5, R6, or R7 is a nitro group), e.g., the compound shown in Figure 13C, designation 13C-9.

[0375] With further reference to Figure 13C and the compound of Formula (I), the formed fourth psilocybin derivative may react to reduce the nitro group to form an amino group and a fifth psilocybin derivative (wherein one or at least one of R2, R4, R5, R6, or R7 is an acetamidyl group, one or at least one of R2, R4, R5, R6, or R7 is a carboxyl ester, and one or at least one of R2, R4, R5, R6, or R7 is an amino group), e.g., the compound shown in Figure 13C, designation 13C-10.

[0376] With further reference to Figure 13C and the compounds of formulas (I) and (II), the fifth psilocybin derivative formed may react with ammonia to form an amide group and a sixth psilocybin derivative of formula (I) (wherein one or at least one of R2, R4, R5, R6, or R7 is an acetamidyl group, one or at least one of R2, R4, R5, R6, or R7 is a carboxyl ester, and one or at least one of R2, R4, R5, R6, or R7 is an amide group), e.g., the compound shown in 13C, designation 13C-11.

[0377] Thus, generally, a reactive psilocybin derivative is provided, and the reactive psilocybin derivative is utilized to react in a chemical reaction that results in the formation of a poly-substituted psilocybin derivative.

[0378] Reactions such as the exemplary reactions shown in Figures 9, 10, 11A-11C, 12A-12D, and 13A-13C may be carried out in any suitable reaction vessel (e.g., tube, bottle). Suitable solvents used may be, for example, water, alcohols (e.g., methanol, ethanol), tetrahydrofuran (THF), dichloromethane, acetone, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), or combinations of such solvents. Suitable temperatures range, for example, from 20°C to 100°C. Furthermore, reaction times may vary. As would be readily apparent to one skilled in the art, reaction conditions may be optimized by, for example, preparing several psilocybin derivative reactant preparations, reacting them in different reaction vessels under different reaction conditions (e.g., different temperatures, use of different solvents, use of different catalysts, etc.), evaluating the resulting polysubstituted psilocybin derivative reaction products, adjusting the reaction conditions, and selecting desirable reaction conditions.

[0379] Turning now to biosynthetic methods for making the disclosed multi-substituted compounds, such methods generally involve enzymatically catalyzing the conversion of a psilocybin derivative precursor compound using a psilocybin biosynthetic enzyme complement to form a multi-substituted psilocybin derivative. The enzymes involved in the psilocybin biosynthetic enzyme complement can vary, as discussed below with reference to certain exemplary enzymes and exemplary compounds shown in Figures 14A, 14B, and 14C.

[0380] Thus, in one aspect, the present disclosure further provides a method for making a polysubstituted psilocybin derivative, the method comprising: [ka] wherein at least one of R2, R4, R5, R6, or R7 is a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and wherein each unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R3 is a hydrogen atom, -CH2-CHNH2COOH, or -CH2-CH2NH2, contacting a psilocybin derivative precursor compound of formula (I) with a catalytic amount of a psilocybin biosynthetic enzyme complement under reaction conditions that allow for enzyme-catalyzed conversion of the psilocybin derivative precursor compound to a compound of formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and R 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group.

[0381] The reaction conditions may be in vitro reaction conditions, in vivo reaction conditions, or a combination thereof. (In vitro synthesis)

[0382] In vitro synthesis generally involves first providing reagents, including precursor psilocybin derivatives and other reagents, in substantially pure form. Thus, the reactants may be provided as particles in substantially pure form, or may be dissolved in substantially pure form in a suitable solvent or diluent, such as water or a buffer solution. The reagents, typically preferably in liquid form (prepared with a diluent, such as water or a buffer solution, if necessary), are then combined and contacted in a suitable reaction vessel (such as a tube, beaker, flask, or, for larger scale use, a tank or reactor). The combined reagents may generally be gently stirred and mixed using a suitable stirring or mixing device, such as a laboratory-scale magnetic stirrer (e.g., manufactured by Fischer Scientific®) or a hand mixer, or an industrial mixer, to form a mixture. The relative and absolute amounts of the reagents may be selected as needed. Absolute amounts are generally based on the desired reaction scale, e.g., laboratory scale (e.g., less than 1 L, less than 100 mL, less than 10 mL, or less than 1 mL) or commercial production scale (e.g., greater than 100 L, greater than 1,000 L, or greater than 10,000 L). Relative and absolute amounts may vary. Thus, for example, in one embodiment, stoichiometric amounts of each precursor psilocybin derivative and substituent-containing compound may be mixed with a catalytic amount of enzyme. If desired, non-stoichiometric amounts of reagents may be selected, e.g., the molar ratio of precursor psilocybin derivative to substituent-containing compound may be 1:0.95, 1:0.9, 1:0.75, 1:1.05, 1:1.1, or 1:1.25.

[0383] As will be understood by those skilled in the art, in terms of molar amounts, enzymes function as catalysts, and because, unlike precursor psilocybin derivatives and substituent-containing compounds, enzymes are not consumed in the reaction, even small amounts of enzyme are sufficient to drive the reaction. Therefore, generally, a catalytic amount can be considered to be at least the minimum amount necessary to convert precursor psilocybin derivatives and substituent-containing compounds to form the desired amount of poly-substituted psilocybin derivatives. Thus, for example, 0.1 to 1,000 units (enzyme units) (e.g., 0.1 unit, 1 unit) of an enzyme may be used. The reaction mixture may contain 10 units, 50 units, 100 units, 250 units, 500 units, or 1,000 units (wherein, as known to those skilled in the art, 1 unit is the amount of enzyme that catalyzes 1 micromole of substituent (i.e., psilocybin precursor derivative) per minute). Furthermore, in vitro reaction conditions may be varied, e.g., temperature may range from about 18°C ​​to about 37°C, and pH may range from about 5.0 to about 8.5. Furthermore, other components, such as diluents (e.g., water or buffer), salts, and pH adjusters, may be included to promote catalysis. In vitro reaction conditions may be adjusted and optimized, for example, by preparing multiple samples and reacting them under different operating conditions (e.g., different temperatures, different pHs, different enzyme amounts, various relative amounts of reagents, etc.) and detecting the polysubstituted psilocybin derivatives formed. (in vivo synthesis)

[0384] In one embodiment, the psilocybin derivative precursor compound and the substituent-containing compound may be contacted with a psilocybin biosynthetic enzyme complement in a host cell, wherein the host cell contains, as operably linked components, the following: (Ii) a nucleic acid sequence that controls expression in a host cell, and (Iii) a chimeric nucleic acid sequence comprising a nucleic acid sequence encoding a psilocybin biosynthetic enzyme complement, wherein the host cell is grown to express the psilocybin biosynthetic enzyme complement and produce a multi-substituted psilocybin derivative compound.

[0385] Suitable chimeric nucleic acid sequences include any nucleic acid sequence having a nucleic acid sequence that controls expression in a host cell operably linked to a sequence encoding one or more of the psilocybin biosynthetic enzyme complement, e.g., tryptophan synthase subunit B polypeptide, tryptophan decarboxylase, N-acetyltransferase, N-methyltransferase, and prenyltransferase, as described in more detail herein below.

[0386] Nucleic acid sequences usable herein that can control the expression of a nucleic acid sequence encoding a biosynthetic enzyme complement in a host cell include any transcription promoter capable of controlling the expression of a polypeptide in a host cell. Generally, for example, when a bacterial host cell is selected according to the present disclosure, a promoter derived from a bacterial cell is used; when a fungal host cell is selected, a fungal promoter is used; and when a plant cell is selected, a plant promoter is used. For example, specific examples usable for expression in yeast cells include galactose-inducible promoters such as the Gal10 / Gal1 promoter, and examples usable for expression in E. coli cells include the beta-galactosidase promoter. Nucleic acid elements capable of controlling expression in a host cell include transcription terminators, enhancers, and the like, all of which may be included in the chimeric nucleic acid sequences of the present disclosure.

[0387] The chimeric nucleic acid sequence may be incorporated into a recombinant expression vector that ensures good expression in a host cell (i.e., is suitable for expression in a host cell). The term "suitable for expression in a host cell" means that the recombinant expression vector contains the chimeric nucleic acid sequence linked to genetic elements necessary for achieving expression in the cell. In this regard, genetic elements contained in an expression vector include a transcription termination region, one or more nucleic acid sequences encoding marker genes, one or more origins of replication, etc. In a preferred embodiment, the expression vector further contains genetic elements necessary for integration of the vector or a portion thereof into the genome of the host cell. When plant cells are used, it also contains T-DNA left and right border sequences that facilitate integration into the nuclear genome of the plant.

[0388] According to the present disclosure, the expression vector may further contain a marker gene. Marker genes used in accordance with the present disclosure include all genes that allow transformed cells to be distinguished from untransformed cells (including all selectable and screenable marker genes). The marker gene may be a disease resistance marker, such as an antibiotic resistance marker for kanamycin or ampicillin, or an auxotrophic marker, such as a Leu marker (Sikorski and Hieter, 1989, Genetics 122(1): 19-27) or a Ura marker (Rose and Winston, 1984, Mol. Gen. Genet. 193(3): 557-560). Screenable markers used to identify transformants by visual inspection include β-glucuronidase (GUS) (U.S. Patent Nos. 5,268,463 and 5,599,670) and green fluorescent protein (GFP) (Niedz et al., 1995, Plant Cell Rep., 14:403).

[0389] A variety of host cells can be used in accordance with the present disclosure. The selected host cell may be capable of naturally producing the psilocybin compound or its derivatives, or the cell may not be capable of naturally producing the psilocybin compound or its derivatives. The host cell may be capable of heterologously expressing a psilocybin biosynthetic enzyme complement through the introduction of a chimeric nucleic acid sequence.

[0390] In some embodiments, the host cell is a microbial cell, such as a bacterial cell or a yeast cell. An exemplary bacterial cell that can be used in accordance with the present invention is an Escherichia coli cell. An exemplary yeast cell that can be used in accordance with the present invention is a Saccharomyces cerevisiae cell or a Yarrowia lipolytica cell.

[0391] In further embodiments, the host cell is a plant cell or an algal cell.

[0392] A variety of techniques and methods exist for engineering host cells to introduce and express nucleic acid sequences (including expression vectors carrying the chimeric nucleic acid sequences of the present disclosure) into cells and are well known to those skilled in the art. Such methods include, for example, cation-based methods (e.g., lithium ion- or calcium ion-based methods), electroporation, gene guns, and glass bead-based methods. As is well known to those skilled in the art, depending on the host cell selected, various methods for introducing nucleic acid components into host cells may be used, and such methods may be optimized for uptake of the nucleic acid components by the host cell, for example, by comparing uptake of the nucleic acid components using various conditions. Detailed guidance may be found, for example, in Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory Press, 2012, Fourth Ed. Chimeric nucleic acids are chimeric nucleic acid sequences that do not occur in nature and may be said to be heterologous to the host cell.

[0393] One exemplary host cell that can be conveniently used is Escherichia coli. Preparation of E. coli vectors may be accomplished using commonly known techniques, such as restriction digestion, ligation, gel electrophoresis, DNA sequencing, polymerase chain reaction (PCR), and other methods. A variety of cloning vectors are available for carrying out the steps necessary to prepare recombinant expression vectors. Among vectors with replication systems, those effective in E. coli include vectors such as pBR322, pUC series vectors, M13 mp series vectors, and pBluescript. Suitable promoter sequences for use in E. coli include, for example, the T7 promoter, T5 promoter, tryptophan (trp) promoter, lactose (lac) promoter, tryptophan / lactose (tac) promoter, lipoprotein (lpp) promoter, and lambda phage PL promoter. Cloning vectors generally contain a marker that allows for selection of transformed cells, e.g., an antibiotic resistance marker such as ampicillin or kanamycin resistance. Nucleic acid sequences may be introduced into such vectors, which may be introduced into E. coli by preparing competent cells, by electroporation, or by other methods known to those skilled in the art. The E. coli are grown in an appropriate medium, such as Luria-Broth medium, and harvested. The recombinant expression vector can be easily recovered from the cells by harvesting and lysing the cells.

[0394] Another exemplary host cell that can be conveniently used is a yeast cell. Exemplary yeast host cells that can be used are yeast cells belonging to the genera Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia, Hansenula, and Yarrowia. In certain exemplary embodiments, the yeast cells are Saccharomyces cerevisiae cells, Yarrowia lipolytica cells, and Pichia pastoris cells.

[0395] There are many vectors for expressing recombinant proteins in yeast host cells. Examples of vectors that can be used in yeast host cells include Yip type vectors, Yep type benders, Yrp type benders, Ycp type benders, pGPD-2, pAO815, pGAPZ, pGAPZα, pHIL-D2, pHIL-S1, pPIC3.5K, pPIC9K, pPICZ, pPICZα, pPIC3K, pHWO10, pPUZZLE, and 2μm plasmid. Such vectors are well known in the art and are described, for example, in Cregg et al., Mol Biotechnol. (2000) 16(1): 23-52. Promoter sequences suitable for use in yeast host cells are also well known and are described, for example, in Mattanovich et al., 1992, Methods Mol. Biol. 2012, 824:329-58 and Romanoest al., 1991, Yeast 8: 423-488. Examples of promoters that can be used in yeast host cells include promoters of glycolytic enzymes, such as triosephosphate isomerase (TPI), phosphoglycerate kinase (PGK), glycerol-3-phosphate dehydrogenase (GAPDH or GAP) and variants thereof, lactase (LAC), galactosidase (GAL), Pichia yeast glucose-6-phosphate isomerase promoter (PPGI), 3-phosphoglycerate kinase promoter (PPGK), glycerolaldehyde phosphate dehydrogenase promoter (PGAP), translation elongation factor promoters, and the like. Marker genes for yeast host cells include promoters of yeast enzymes such as PTEF, Saccharomyces cerevisiae enolase (ENO-1), Saccharomyces cerevisiae galactokinase (GAL1), Saccharomyces cerevisiae alcohol dehydrogenase / glyceraldehyde-3-phosphate dehydrogenase (ADH1, ADH2 / GAP), Saccharomyces cerevisiae triosephosphate isomerase (TPI), Saccharomyces cerevisiae metallothionein (CUP1), Saccharomyces cerevisiae phosphoglycerate kinase (PGK), and maltase gene promoter (MAL). Marker genes for use in yeast host cells are also well known in the art.Thus, antibiotic resistance markers such as ampicillin resistance markers may also be used in yeast, as well as marker genes that provide gene function for essential nutrients, such as leucine (LEU2), tryptophan (TRP1 and TRP2), uracil (URA3, URA5, URA6), histidine (HIS3), etc. Methods for introducing vectors into yeast host cells may be found, for example, in Kawai et al., 2010, Bioeng. Bugs 1(6): 395-403.

[0396] Further, preparation of expression vectors and their introduction into host cells including E. coli cells, yeast cells, and other host cells may be found, for example, in Sambrook et al. Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory Press, 2012, Fourth Ed.

[0397] Referring now to Figures 14A-14G, exemplary biosynthetic pathways for producing the polysubstituted psilocybin derivative compounds of the present application are shown. In particular, for example, Figure 14A illustrates an exemplary method for the biosynthetic production of an S14,S26 polysubstituted psilocybin derivative. Figure 14B illustrates, by way of further example, a method for the biosynthetic production of an additional exemplary S14,S26 polysubstituted psilocybin derivative. Figure 14C illustrates, by way of further example, a method for the biosynthetic production of an additional exemplary S15,S26 polysubstituted psilocybin derivative. Figure 14D illustrates, by way of example, a method for the biosynthetic production of an exemplary S14,S25 polysubstituted psilocybin derivative. Figure 14E illustrates, by way of example, a method for the biosynthetic production of an exemplary S14,S27 polysubstituted psilocybin derivative. Figure 14F illustrates, by way of example, a method for the biosynthetic production of an exemplary S15,S26 polysubstituted psilocybin derivative. Figure 14G illustrates, by way of further example, a biosynthetic method for the preparation of exemplary S15,S27 polysubstituted psilocybin derivatives. In addition to the pathways, enzymes, and compounds shown in Figures 14A-14G, other biosynthetic pathways are available for producing other polysubstituted psilocybin derivative compounds (wherein two or more of the carbon atoms R2, R4, R5, R6, or R7 have a substituent selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group), such as the polysubstituted compounds shown in Figures 3A-3L, 4A-4I, 5A-5I, 6A-6I, and 7A-7F, as well as compounds of Formula (I): [ka] wherein at least two of R2, R4, R5, R6, or R7 are substituents independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and each of unsubstituted R2, R5, R6, or R7 is a hydrogen atom; and when R4 is not substituted with any of the above substituents, R4 is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group; and 3a and R 3b are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group, and R 3c is a hydrogen atom or a carboxyl group). Those skilled in the art will be able, with reference to the teachings herein, including the examples shown in Figures 14A-14G, to select suitable psilocybin precursor compounds and psilocybin biosynthetic enzyme complements for biosynthetically producing the poly-substituted psilocybin derivative compounds of the present application.

[0398] Thus, with reference to FIG. 14A and the psilocybin derivative precursor compound (LVII), the psilocybin biosynthetic enzyme complement may include, for example, the following: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) A tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from the nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f). A psilocybin derivative precursor compound of formula (I) (wherein two of R2, R4, R5, R6, or R7 are independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom) may be used. A first polysubstituted psilocybin derivative compound of formula (I) (wherein R 3c is a carboxy group). For example, a psilocybin derivative precursor compound may be formed having the formula (LVIII): [ka] (wherein R4 is a hydroxy group and R6 is a chlorine atom), and may be a compound of the following formula (LV): [ka] The first-formed poly-substituted psilocybin derivative compound may be formed in an in vivo or in vitro reaction catalyzed by a tryptophan synthase subunit B polypeptide.

[0399] With further reference to FIG. 14A, the psilocybin biosynthetic enzyme complement decarboxylates the R—CH—CHNHCOOH group of the first polysubstituted psilocybin derivative compound to form the compound of formula (I) (R 3a and R 3b and each is a hydrogen atom), the tryptophan decarboxylase being capable of reacting with the following: (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0400] With further reference to FIG. 14A, in one exemplary embodiment, the psilocybin derivative precursor compound has the formula (LVIII): [ka] (wherein R4 is a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom), and may be a compound of the following formula (LIX): [ka] (wherein R4 is a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom), the first polysubstituted psilocybin may be decarboxylated to form a second polysubstituted psilocybin, for example, a compound of the following formula (XXII), (XXVI), (XXIX), (LII), or (LIV): [ka] A second polysubstituted psilocybin derivative may be formed having the formula:

[0401] The psilocybin biosynthetic enzyme complement can be, for example, a compound represented by formula (I) (R 3a is a hydrogen atom, and R3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I). (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0402] With further reference to FIG. 14A, in one exemplary embodiment, the psilocybin derivative precursor compound has the formula (LVIII): [ka] wherein R4 is an acetamidyl group, a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom, and the first polysubstituted psilocybin derivative compound may be a compound represented by the following formula (LIX): [ka] (wherein R4 is an acetamidyl group, a fluorine atom, an amino group, or a hydroxy group, and R6 is a fluorine atom, an amino group, a nitrile group, or a bromine atom) may be formed. The first polysubstituted psilocybin derivative may then be decarboxylated to form a compound of the following formula (LX): [ka] The second polysubstituted psilocybin derivative may then be acetylated to form a third polysubstituted psilocybin derivative, for example, having the following formula (IX), (X), (XVIII), (XXI), (XXV), or (XXVIII): [ka] A third polysubstituted psilocybin derivative of the formula:

[0403] The psilocybin biosynthetic enzyme complement according to the present invention methylates the R3 amino group of R3 to form a psilocybin analog of formula (I) (R 3a and R 3b are methyl groups, or R 3a is a hydrogen atom, and R 3b The psilocybin derivative may further comprise an N-methyltransferase that forms a fourth polysubstituted psilocybin derivative (wherein N-methyltransferase is a methyl group). (a) SEQ ID NO: 11 and SEQ ID NO: 13; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0404] With further reference to FIG. 14A, in one further embodiment, the psilocybin derivative precursor compound has the formula (LVIII): [ka] (wherein R4 is an amino group or a hydroxy group, and R6 is a chlorine atom, a nitrile group, or a bromine atom), and may be a compound of the following formula (LIX): [ka] wherein R4 is an amino group or a hydroxy group, and R6 is a chlorine atom, a nitrile group, or a bromine atom. The first polysubstituted psilocybin derivative compound may be decarboxylated to form a second polysubstituted psilocybin derivative having the following formula (LX): [ka] It has. The third polysubstituted psilocybin derivative may be methylated to produce a fourth polysubstituted psilocybin derivative, for example, one of the following formulas (XXVII), (XL), or (LIII): [ka] quaternary polysubstituted psilocybin derivative compounds of the formula:

[0405] In accordance with the present specification, in further exemplary embodiments, the psilocybin biosynthetic enzyme complement is: (a) SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, and SEQ ID NO: 21; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, and SEQ ID NO: 22; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, and SEQ ID NO: 22; and (g) The prenyltransferase may be encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0406] Referring now to FIG. 14B, in a further embodiment, the psilocybin derivative precursor compound has the formula (LXI): [ka] (wherein R4 is a hydroxy group), a first poly-substituted psilocybin derivative compound may be formed, the first poly-substituted psilocybin derivative compound having, for example, the following formula (L): [ka] may have

[0407] With further reference to FIG. 14B, in a further embodiment, the psilocybin biosynthetic enzyme complement decarboxylates the R—CH—CHNH—COOH group to form a psilocybin analog of formula (I) (R 3a and R 3b and each are a hydrogen atom), the tryptophan decarboxylase forming a second polysubstituted psilocybin derivative comprising: (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0408] In a further exemplary embodiment, the psilocybin biosynthetic enzyme complement methylates the R3 amino group of R3 to form a psilocybin analog of formula (I) (R 3a and R 3b are methyl groups, or R 3a is a hydrogen atom, and R 3b The psilocybin derivative may further comprise an N-methyltransferase that forms a fourth polysubstituted psilocybin derivative (wherein N-methyltransferase is a methyl group). (a) SEQ ID NO: 11 and SEQ ID NO: 13; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 14; and (g) Encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0409] With further reference to FIG. 14B, in one exemplary embodiment, the psilocybin derivative precursor compound has the following formula (LXI): [ka] (wherein R4 is a propionyloxy group or an acetoxy group), and may be a compound of the following formula (LIX): [ka] The first polysubstituted psilocybin derivative compound is decarboxylated to form a compound of the formula: [ka] where R4 is a propionyloxy or acetoxy group and R6 is a prenyl group) may be formed. The second polysubstituted psilocybin derivative is methylated and has the following formula (XLI) or (XLII): [ka] A third polysubstituted psilocybin derivative of the formula:

[0410] With further reference to Figure 14B, dimethylallyl pyrophosphate (DMAPP) may be used as a substituent-containing compound to prenylate the S14 psilocybin derivative precursor compound. Other prenyl-containing compounds, such as geranyl pyrophosphate (GPP), farnesyl pyrophosphate (FPP), and geranylgeranyl pyrophosphate (GGPP), may alternatively be used as the substituent-containing compound. As further shown in Figure 14B, DMAPP itself may optionally be formed biosynthetically (in vitro or in vivo) from dimethylallyl alcohol (DMAOH) using phosphatase and isopentenyl phosphate kinase as catalytic enzymes, as further described in Couillaud et al., 2019, ACS Omega, 4, 7838-7859.

[0411] Referring now to FIG. 14C, in a further embodiment, the psilocybin derivative precursor compound has the formula (LXII): [ka] (wherein R5 is a chlorine or fluorine atom), and may be a compound of the following formula (LXIII): [ka] The first polysubstituted psilocybin derivative may be decarboxylated to form a first polysubstituted psilocybin derivative compound of the following formula (XXXVI) or (XXXVIII): [ka] A second polysubstituted psilocybin derivative compound of the formula:

[0412] In a further exemplary embodiment, the psilocybin biosynthetic enzyme complement is represented by the formula (I) (R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0413] With further reference to FIG. 14C, in a further embodiment, the psilocybin derivative precursor compound has formula (LXII): [ka] (wherein R5 is a chlorine or fluorine atom), and may be a compound of the following formula (LXIII): [ka] wherein R5 is a chlorine or fluorine atom and R6 is a prenyl group, the first polysubstituted psilocybin derivative compound may be decarboxylated to form a compound of the following formula (LXIV): [ka] wherein R5 is a chlorine or fluorine atom and R6 is a prenyl group, the second polysubstituted psilocybin derivative compound may be acetylated to form a compound of the following formula (XXXV) or (XXXVII): [ka] A third polysubstituted psilocybin derivative of the formula:

[0414] Further referring to Figure 14C, similar to the biosynthetic pathway shown in Figure 14B, dimethylallyl pyrophosphate (DMAPP) may be used as a substituent-containing compound to prenylate the S15 psilocybin derivative precursor compound shown in Figure 14C.

[0415] Referring now to FIG. 14D and the psilocybin derivative precursor compound of formula (LVII), the psilocybin biosynthetic enzyme complement may include, for example, the following: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) a tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0416] A first psilocybin derivative precursor compound of formula (I) (wherein two of R2, R4, R5, R6, or R7 are substituents independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom) may be used. A first polysubstituted psilocybin derivative compound of formula (I) (wherein R 3c is a carboxyl group). For example, a psilocybin derivative precursor compound may be formed having the following formula (LXV): [ka] (wherein R4 is a hydroxy group and R5 is a prenyl group), and may be a compound of the following formula (LI): [ka] A first polysubstituted psilocybin derivative compound of the formula:

[0417] The psilocybin biosynthetic enzyme complement decarboxylates the R-CH-CHNHCOOH group to form the compound of formula (I) (R 3a and R 3b and each are a hydrogen atom), wherein the tryptophan decarboxylase is (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0418] With further reference to FIG. 14D, for example, a psilocybin derivative precursor compound may be represented by the following formula (LXV): [ka] (wherein R4 is a fluorine atom and R5 is a nitrile group), and may be a compound of the following formula (LXIX): [ka] The first poly-substituted (psilocybin) derivative compound may then be decarboxylated to form a first poly-substituted psilocybin derivative compound of the following formula (XIX): [ka] A second polysubstituted psilocybin derivative compound of the formula:

[0419] In a further embodiment, the psilocybin biosynthetic enzyme complement is represented by formula (I)(R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0420] With further reference to FIG. 14D, for example, a psilocybin derivative precursor compound may be represented by the following formula (LXV): [ka] (wherein R4 is a fluorine atom and R5 is a hydroxy group or a nitrile group), and may be a compound of the following formula (LXIX): [ka] wherein R4 is a fluorine atom and R5 is a hydroxy group or a nitrile group. The first polysubstituted psilocybin derivative compound may be decarboxylated to form a compound of the following formula (LXXII): [ka] where R4 is a fluorine atom and R5 is a hydroxy group or a nitrile group. A second polysubstituted psilocybin derivative compound is then acetylated and has the following formula (XVII) or (XX): [ka] A third polysubstituted psilocybin derivative of the formula:

[0421] Referring now to FIG. 14E, the psilocybin biosynthetic enzyme complement is as follows: (a) SEQ ID NO: 1; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO:2; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 2; and (g) a tryptophan synthase subunit B polypeptide encoded by a nucleic acid selected from a nucleic acid sequence that hybridizes under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f), wherein in the psilocybin derivative precursor compound of formula (LVIII), two of R2, R4, R5, R6, or R7 are substituents independently selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, and R3 is a hydrogen atom, thereby forming a first polysubstituted psilocybin derivative compound of formula (I), and R 3c is a carboxyl group. For example, a psilocybin derivative precursor compound may be represented by the following formula (LXVI): [ka] (wherein R4 is a hydroxy group and R7 is a prenyl group), and may be a compound of the following formula: XLIX): [ka] A first polysubstituted psilocybin derivative compound of the formula:

[0422] In a further embodiment, the psilocybin biosynthetic enzyme complement decarboxylates the R—CH—CHNH—COOH group to form a psilocybin analog of formula (I) (R 3a and R 3b and each are a hydrogen atom), wherein the tryptophan decarboxylase is (a) SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7; (b) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a); (c) a nucleic acid sequence that is substantially identical to any one of the nucleic acid sequences of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to any one of the nucleic acid sequences of (a); (e) a nucleic acid sequence encoding a polypeptide having any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; (f) a nucleic acid sequence encoding a functional variant of any one of the amino acid sequences set forth in SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:8; and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0423] With further reference to FIG. 14E, the psilocybin derivative precursor compound has the following formula (LXVI): [ka] (wherein R4 is a fluorine atom and R7 is a nitrile group), and may be a compound of the following formula (LXX): [ka] The first polysubstituted psilocybin derivative compound may be decarboxylated to form a compound of the following formula (XXIV): [ka] A second polysubstituted psilocybin derivative compound of the formula:

[0424] In a further embodiment, the psilocybin biosynthetic enzyme complement is represented by formula (I)(R 3a is a hydrogen atom, and R 3b is an acetyl group) to form a third polysubstituted psilocybin of formula (I), wherein the N-acetyltransferase is (a) SEQ ID NO: 9; (b) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a); (c) a nucleic acid sequence that is substantially identical to the nucleic acid sequence of (a) without alterations of the genetic code; (d) a nucleic acid sequence complementary to the nucleic acid sequence of (a); (e) a nucleic acid sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10; (f) a nucleic acid sequence encoding a functional variant of the amino acid sequence set forth in SEQ ID NO: 10, and (g) may be encoded by a nucleic acid sequence selected from nucleic acid sequences that hybridize under stringent conditions to any one of the nucleic acid sequences set forth in (a), (b), (c), (d), (e), or (f).

[0425] With further reference to FIG. 14E, the psilocybin derivative precursor compound has the following formula (LXVI): [ka] (wherein R4 is a fluorine atom or a chlorine atom, and R7 is a prenyl group or a nitrile group), and may be a compound of the following formula (LXX): [ka] The first polysubstituted psilocybin derivative compound may be decarboxylated to form a polysubstituted psilocybin derivative compound of the following formula (LXXIII): [ka] After acetylation, a second polysubs...

Claims

1. A compound of formula (I) or a salt thereof: 【Chemistry 1】 R ２ , R ４ , R ５ , R ６ , or R ７ at least two of R are independently selected from at least two of (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group; and unsubstituted R ２ , R ５ , R ６ , or R ７ Each of R is a hydrogen atom; ４ When R is not substituted by any of the above substituents, ４ is a hydrogen atom, an O-alkyl group, an O-acyl group, or a phosphate group, and R ３ａ and R ３ｂ are each independently a hydrogen atom, an alkyl group, a cycloalkyl group, an acyl group, or an aryl group; R ３ｃ is a hydrogen atom or a carboxyl group A compound or a salt thereof.

2. A compound according to claim 1, wherein R 2 is hydrogen, and only two of R 4 , R 5 , R 6 , or R 7 are substituted, the first substituent is selected from (i) a halogen atom, (ii) a prenyl group, and (iii) a nitrile group, and the second substituent is selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, preferably selected from (i) a halogen atom, (ii) a prenyl group, (iii) a hydroxy group, (iv) a nitro group, (v) an amino group or an N-substituted amino group, and (vi) a carboxyl group or a carboxylic acid derivative, and the first and second substituents are different groups.

3. A compound according to claim 1 or 2, wherein R2 is hydrogen and only two of R4, R5, R6, or R7 are substituted, and the first substituent is a halogen atom and the second substituent is selected from (i) an amino group or an N-substituted amino group, (ii) a nitrile group, (iii) a nitro group, (iv) a hydroxy group, and (v) a prenyl group, or the first substituent is a prenyl group and the second substituent is selected from (i) a carboxyl group or a carboxylic acid derivative, (ii) a halogen, and (iii) a hydroxy group, or the first substituent is a nitrile group and the second substituent is an amino group or an N-substituted amino group.

4. A compound according to any one of claims 1 to 3, wherein R 2 is hydrogen, and only two of R 4 , R 5 , R 6 , or R 7 are substituted, the first substituent is a halogen atom, and the second substituent is selected from (i) a halogen atom, (ii) a hydroxy group, (iii) a nitro group, (iv) a glycosyloxy group, (v) an amino group or an N-substituted amino group, (vi) a carboxyl group or a carboxylic acid derivative, (vii) an aldehyde or ketone group, (viii) a prenyl group, and (ix) a nitrile group, preferably selected from (i) a prenyl group, and (ii) an aldehyde or ketone group, and the first and second substituents are not identical.

5. A compound according to claim 1, wherein R 2 is hydrogen, and only two of R 4 , R 5 , R 6 , or R 7 are substituted, and the first and second substituents are selected from (i) a halogen atom, (ii) a prenyl group, and (iii) an aldehyde or ketone group, respectively, and the first and second substituents are not identical.

6. A compound according to claim 1, wherein R 4 and R 6 are selected from two of (i) a halogen atom, (ii) a prenyl group, and (iii) an aldehyde or ketone group, the first and second substituents are not identical, and R 2, R 5, and R 7 are hydrogen atoms.

7. A compound described in any one of claims 1 to 6, wherein R 5 and R 6 are selected from two of (i) a halogen atom, (ii) a prenyl group, and (iii) an aldehyde or ketone group, the first and second substituents are not identical, and R 2, R 4, and R 7 are hydrogen atoms.

8. A compound according to any one of claims 1, 2, 4, 5 and 6, wherein R 4 and R 6 are (i) a halogen atom and an aldehyde or ketone group, respectively, and R 2 , R 5 and R 7 are hydrogen atoms.

9. The compound according to claim 1, wherein R 5 and R 6 are (i) a halogen atom and a prenyl group, respectively, and R 2 , R 4 , and R 7 are hydrogen atoms.

10. The compound according to claim 1, wherein the halogen atom is a fluorine atom.

11. A compound described in any one of claims 1 to 10, wherein R 3a and R 3b are hydrogen atoms and acyl groups, and R 3c is a hydrogen atom, or R 3a and R 3b are independently selected from alkyl groups such as methyl groups, ethyl groups, propyl groups, or longer chain alkyl groups, and hydrogen atoms, and R 3c is a hydrogen atom.

12. The compound according to claim 1, wherein compound (I) is represented by the following chemical formula (IX), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXXI), (XXII), (XXIII), (XXIV), (XXV), (XXVI), (XXVII), (XXVIII), (XXIX), (XXX), (XXXI), (XXXII), (XXXIII), (XXXIV), (XXXV), (XXXVI), (XXXVII), (XXXVII), (XXVII), (XXIX), (XXX), (XXXX), (XXXII), (XXXIII), (XXXIV), (XXXV), (XXXVI), (XXXVII), (XXXVII), (XX ... XXXVIII), (XXXIX), (XL), (XLI), (XLII), (XLIII), (XLIV), (XLV), (XLVI), (XLVIII), (XLVIII), (XLIX), (L), (LI), (LII), (LIII), (LIV) : 【Chemistry 2-1】 【Chemistry 2-2】 【Chemistry 2-3】 【Chemistry 2-4】 【Chemistry 2-5】 【Chemistry 2-6】 The compound is selected from the group consisting of the compounds:

13. A compound according to claim 1, wherein R2 is hydrogen, and only two of R4, R5, R6, or R7 are substituted, the first substituent is a halogen atom, and the second substituent is selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, (vi) an aldehyde or ketone group, (vii) a prenyl group, and (viii) a nitrile group.

14. A compound according to claim 1, wherein R 2 is hydrogen and only two of R 4 , R 5 , R 6 , or R 7 are substituted, the first substituent is selected from (i) a halogen atom, (ii) a prenyl group, and (iii) a nitrile group, and the second substituent is selected from (i) a hydroxy group, (ii) a nitro group, (iii) a glycosyloxy group, (iv) an amino group or an N-substituted amino group, (v) a carboxyl group or a carboxylic acid derivative, and (vi) an aldehyde or ketone group.

15. The compound according to claim 1, wherein R 2 is hydrogen, only two of R 4 , R 5 , R 6 , or R 7 are substituted, the first substituent is a halogen atom, and the second substituent is a prenyl group.

16. A compound according to any one of claims 1 to 15, wherein the compound is at least about 95% (w / w) pure, preferably at least about 97% (w / w) pure, and more preferably at least about 99% (w / w) pure.

17. 1. A pharmaceutical formulation comprising: A composition comprising an effective amount of a compound according to any one of claims 1 to 16 together with a pharma- ceutically acceptable excipient, diluent, or carrier. Pharmaceutical preparations.

18. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 15 for use in the treatment of a psychiatric disorder, The pharmaceutical formulation is administered to a subject in need thereof in an effective amount to treat the psychiatric disorder in the subject. Pharmaceutical preparations.

19. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 15 for use in the treatment of a psychiatric disorder, The disorder is 5-HT ２Ａ Receptor-Mediated Disorders or 5-HT １Ａ It is a receptor-mediated disorder. Pharmaceutical preparations.

20. A compound according to any one of claims 1 to 15 for use in a pharmaceutical formulation for the treatment of a psychiatric disorder, said pharmaceutical formulation being administered to a subject in need thereof in a dosage of from about 0.001 mg to about 5,000 mg.

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