Scalable synthetic routes to psilocin and psilocybin.

Abstract

This application relates to a novel, cost-effective, practical, and scalable synthetic route for the synthesis of psilocybin (compound of Formula I) and its major metabolite, psilocin (compound of Formula II), and their pharmaceutically acceptable salts. These compounds are used to treat or prevent mental health disorders, such as major depressive disorder, anxiety, and addictive disorders, among other CNS disorders. JPEG2023535742000100.jpg43132

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 056,058, filed July 24, 2020, the contents of which are incorporated herein by reference in their entirety.

[0002] Field of Disclosure The present invention relates to a scalable process for preparing psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine) and its major metabolite, psilocin (4-hydroxy-N,N-dimethyltryptamine), as well as pharmaceutically acceptable salts thereof. [Background technology]

[0003] Psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine, also known as psilocin phosphate) has the chemical formula C 12 H 17 It has N2O4P, an exact molecular weight of 284.092594 g / mol, three hydrogen bond donors, five hydrogen bond acceptors, and a polar surface area (PSA) or topological polar surface area (TPSA) of 85.8 Å. 2 (Formula I). When consumed via the oral route, psilocybin is rapidly dephosphorylated to psilocin by alkaline phosphatase in the intestine [Dinis-Oliveira, RJ Drug Metab. Rev. 2017, 49, 84-91]. Psilocin (4-hydroxy-N,N-dimethyltryptamine), also known as 4-OH-DMT, is the most prominent active metabolite of psilocybin and is capable of penetrating the blood-brain barrier (BBB) and targeting 5-HT, among other targets. 2A It interacts with receptors to produce hallucinogenic effects. Its chemical formula is C 12 H 16 N2O has an exact molecular weight of 204.126263 g / mol, two hydrogen bond donors, two hydrogen bond acceptors, and a 39.3 Å 2 (Formula II).

[0004] The chemical structures of psilocybin and psilocin are fairly simple and achiral, and are closely related to the neurotransmitter serotonin [5-hydroxytryptamine (5-HT)].

[0005] Psilocybin has been shown to have many potential pharmaceutical therapeutic values, for example, in hallucination-assisted therapy for psychiatric diseases and disorders, opioid use disorder (OUD), sleep disorders, anxiety disorders, major depressive disorder, and cancer-related psychiatric distress, and as a hallucinogen. Its therapeutic value, coupled with its increasing popularity worldwide as a recreational drug, warrants the need for more rigorous research with psilocybin. As a result, scientific research efforts are increasing to create more cost-effective and reproducible methods for the production of psilocybin. Specifically, there is a strong demand for investigational substances that are pure enough for human use and can be prepared in a relatively economical manner.

[0006] Currently, psilocybin can be produced using one of three different methods: (i) by a biological process such as fermentation, (ii) by extraction from magic mushrooms, or (iii) by organic synthesis.

[0007] Psilocybin is biosynthesized in certain mushrooms and can therefore be extracted from natural sources. However, this method relies on a supply of mushrooms, whose production can be inconsistent and difficult to control. Furthermore, because psilocybin readily decomposes into its main metabolite, psilocin, which has high polarity, it is extremely difficult to produce psilocybin in sufficient and pure quantities for use in human trials from naturally occurring mushroom extracts due to its stability. Therefore, this method is not a useful drug for production. Additionally, reported psilocybin yields from natural sources are typically very low, e.g., 0.85% of the dry mass for Psilocybe serbica, approximately 1.0% for P. semilanceata, and approximately 1.5-1.8% for P. azurescens (Tyls, T. et al. Eur. Neuropsychopharm. 2014, 24, 342-356; Hoffmeister, Chem. Eur. J. 2019, 25, 897-903). Therefore, this method is not a viable commercial production method for psilocybin for therapeutic use.

[0008] Separately, biosynthetic psilocybin production using enzymes derived from psilocybin-containing fungi has recently been identified and is outlined in four steps (Scheme 1) via biochemical conversion from L-tryptophan (A), involving several enzymatic reactions: decarboxylation yields tryptamine (B); dimethylation of the amine in the ethylamine side chain yields N,N-dimethyltryptamine (C); oxidative 4-hydroxylation yields 4-hydroxytryptamine, psilocin (II); and subsequent O-phosphorylation yields psilocybin (I) [Fricke, J. et al., Angew. Chem., Int. Ed. 2017, 56, 12352-12355].

[0009] [ka]

[0010] Recent analysis of isolated enzymes indicates that O-phosphorylation is the third step in P. cubensis. The sequence of intermediate enzymatic steps has been shown to involve four distinct enzymes (PsiD, PsiH, PsiK, and PsiM) in P. cubensis and P. cyanescens, although the biosynthetic pathway may differ between species. These enzymes are encoded in gene clusters in Psilocybe, Panaeolus, and Gymnopilus [Fricke J. et al. (Angew. Chem., 2017, 56(40): 12352-12355; Reynolds H, et al., Evolution Letters. 2018, 2 (2): 88-101].

[0011] In more recent work, another modular biosynthetic psilocybin production platform has been developed in the model microorganism, Escherichia coli. Efforts to optimize and improve pathway performance using multiple genetic optimization techniques were evaluated, resulting in a 3.2-fold improvement in psilocybin titer. Further enhancements to this genetically superior strain were achieved through fermentation optimization, ultimately resulting in a fed-batch fermentation study with a psilocybin production titer of 1.16 g / L for use in ongoing clinical trials [Adams et al. Metabolic Engineering, 2019, 56, 111-119].

[0012] Alternatively, the biosynthetic process may also begin with 4-hydroxy-L-tryptophan, which is decarboxylated via PsiD to yield 4-hydroxytryptamine, and psilocybin biosynthesis continues from this point as per Scheme 1 [Fricke, J. et al., Angew. Chem., Int. Ed. 56, 2017, 12352-12355].

[0013] Similarly, two enzymes from Psilocybe cubensis were recently reported to perform a two-step cascade that prepares psilocybin for oxidative oligomerization, leading to a blue product. The phosphatase PsiP removes the 4-O-phosphate group to give psilocin, while PsiL oxidizes its 4-hydroxyl group. The PsiL reaction occurs in situ. 13 The oxidative coupling of psilocybin residues, monitored by C-NMR spectroscopy, indicated that it occurred primarily via C-5. MS and IR spectroscopy demonstrated the formation of a heterogeneous mixture with a predominance of psilocylic 3- to 13-mers, suggesting multiple oligomerization pathways depending on oxidizing power and substrate concentration. This result also suggests that the phosphate ester of psilocybin serves a reversible protective function [Claudius L. et al., Angew. Chem. Int. Ed. 2020, 59, 1450-1454].

[0014] International Application Publication No. WO 2019 / 180309 discloses an enzymatic pathway for the biosynthesis of psilocybin. This pathway uses the amino acid L-tryptophan as the initial substrate. L-tryptophan is converted to tryptamine and CO in the first enzymatic reaction catalyzed by the PsiD enzyme. Tryptamine is converted to 4-hydroxytryptamine in the second enzymatic reaction catalyzed by the PsiH enzyme, where oxygen is used in the reaction to form a hydroxyl group. 4-hydroxytryptamine is converted to norbaeocystin in the third enzymatic reaction catalyzed by the PsiK enzyme, where adenosine triphosphate (ATP) is used as the phosphate group donor. Norbaeocystin is converted to baeocystin and ultimately psilocybin in the fourth and fifth enzymatic reactions catalyzed by the PsiM enzyme, where S-adenosylmethionine (SAM) is used as the methyl group donor. Psilocybin can be converted to psilocybin in a reaction catalyzed by natural host phosphatases, or spontaneously, and psilocin can be rephosphorylated by the PsiK enzyme to form psilocybin again.

[0015] Psilocybin can also be prepared synthetically. In fact, its original synthesis was reported by Hofmann and coworkers as shown in Scheme 2 [Hofmann, A. et al. Experientia 1958, 14, 397-399; Helv. Chim. Acta 1959, 42, 2073-2103]. The synthesis begins with benzylation of 4-hydroxyindole (E), resulting in benzyl-protected 4-hydroxyindole (F), which is treated with oxalyl chloride and dimethylamine to produce 2-(4-(benzyloxy)-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide (G). The product is subsequently reduced with lithium aluminum hydride to give benzyl-protected psilocin (H). Deprotection with palladium on carbon under hydrogen gas affords psilocin (II). This product is then phosphorylated with O,O-dibenzylphosphoryl chloride to give benzyl-protected psilocybin (IIa). Debenzylation under hydrogen gas using palladium on carbon gives the final product, psilocybin (I) [Hofmann, A. et al. Experientia 1958, 14, 397-399; Helv. Chim. Acta 1959. 42, 2073-2103].

[0016] [ka]

[0017] Hofmann's synthesis has since been modified by various groups, who have made numerous contributions to the process to improve overall purity and yield. For example, it was recently shown that during the synthesis of psilocybin (1), a by-product is formed by the spontaneous intramolecular migration of one of the benzyl groups on the O,O-dibenzylphosphate derivative (IIa) to form the corresponding zwitterionic N,O-dibenzylphosphate derivative (IIb) (Scheme 3) [Shirota, O. et al., J. Nat. Prod. 2003, 66, 885-887].

[0018] [ka]

[0019] An improved procedure for achieving O-phosphorylation of 4-hydroxy-N,N-dimethyltryptamine, psilocin (II), has been reported, utilizing the reaction between the O-lithium salt of the indole moiety, n-butyllithium as a base, and tetra-O-benzyl pyrophosphate. The O-benzyl group is then removed by catalytic hydrogenation over palladium / carbon to yield psilocybin. Given the difficulties encountered in the preparation of psilocybin, it has been suggested that 4-acetoxy-N,N-dimethyltryptamine may be a useful alternative for pharmacological studies. The latter has been obtained after catalytic O-debenzylation of 4-benzyloxy-N,N-dimethyltryptamine in the presence of acetic anhydride and sodium acetate [Nichols, D. et al., Synthesis 1999, 935-938].

[0020] A slightly improved process for the preparation of psilocybin (I) based on the same synthetic route developed by Hoffmann and coworkers, but using acetoxy-indole instead of benzyloxy-indole as the starting material, is disclosed in International Application Publication No. WO 2019 / 073379.

[0021] Similarly, a recent small-scale synthesis of psilocybin (I) in five steps without the use of chromatography or aqueous workup has been described, resulting in an overall yield of 23% [Sherwood AM Synthesis 2020, 52, 688-694].

[0022] A kilogram-scale synthesis of psilocin (I) has also been disclosed, in which psilocin is directly phosphorylated with phosphorus oxychloride without the use of protecting groups [Sherwood et al., ACS Omega 2020, 5, 27, 16959-16966].

[0023] Another synthesis of 4-hydroxyindole from indole via 4-iodoindole using thallium acetate has been reported, and this has been used to prepare psilocin (II) [Yamada, F. et al., M. Heterocycles 1998, 49, 451].

[0024] A three-step method for preparing silosin (II) from N-tert-butoxycarbonyl-2-iodo-3-methoxyaniline (J) without using thallium salts has also been reported (Scheme 4).

[0025] [ka]

[0026] The key step in the above method is the formation of the indole core via palladium-catalyzed cyclization. The two fragments required for cyclization are (J) and alkyne (K). Intermediate (J) was prepared from Boc-protected 3-methoxyaniline via directed lithiation and iodination according to literature procedures [Snieckus V. Chem. Rev. 1990, 90, 879]. Preparation of (K) was achieved from 3-butyn-1-ol as previously reported [Smith, AL GB23.28941, 1999]. Tosylation, displacement with N,N-dimethylamine, and treatment with n-butyllithium and trimethylsilyl chloride afforded the required alkyne. The key palladium-catalyzed cyclization step was achieved using Pd(OAc)2, triphenylphosphine, tetraethylammonium chloride, and N,N-diisopropylethylamine in DMF at 80 °C for 48 h.

[0027] To complete the synthesis of psilocin, the Boc and trimethylsilyl groups of (L) were cleaved by treatment with neat TFA to give (M), which was subjected to O-demethylation with boron tribromide to give psilocin (II) [Scammells PJ et al. Org. Lett., 5, 6, 2003].

[0028] The main drawbacks of this latter synthesis are the expensive and corrosive iodine intermediate and the handling of toxic and expensive butyllithium. Palladium acetate is very expensive. Also, the 3-butyn-l-ol used in this method is expensive and not readily available on a commercial scale. The purity of psilocybin's precursor, psilocin, is low and requires multiple column chromatography processes for purification. Therefore, this method is not viable for commercial production of psilocybin.

[0029] In view of the difficulties in commercializing the above-described methods for preparing psilocin and psilocybin, there is a need to develop a simple and economical method for the commercial production of psilocin and / or psilocybin. Summary of the Invention [Problem to be solved by the invention]

[0030] Applicants have developed a novel, cost-effective, practical, and scalable route for the preparation of psilocybin (compound of formula I), which is (4-phosphoryloxy-N,N-dimethyltryptamine), and its major metabolite, psilocin (compound of formula II), which is (4-hydroxy-N,N-dimethyltryptamine). [Means for solving the problem]

[0031] Thus, the present application includes a process for preparing psilocin (a compound of formula II).

[0032] [ka]

[0033] The process is: Unprotected 4-hydroxyindole (compound of formula III)

[0034] [ka]

[0035] with oxalyl chloride to provide a compound of formula (IV);

[0036] [ka]

[0037] reacting the compound of formula (IV) with dimethylamine (HN(CH3)2) to provide a compound of formula V;

[0038] [ka]

[0039] and The method comprises reducing the compound of formula (V) with a reducing agent to provide psilocin (compound of formula II).

[0040] The present application also provides a method for preparing psilocybin (compound of formula I), comprising:

[0041] [ka]

[0042] Psilocin (compound of formula II)

[0043] [ka]

[0044] in the presence of a base (a) di-tert-butyl phosphite

[0045] [ka]

[0046] by reacting with, or Silosin (b) chlorodi-tert-butylphosphite

[0047] [ka]

[0048] to provide a compound of formula (VI);

[0049] [ka]

[0050] and Hydrolyzing the compound of formula VI to provide psilocybin (the compound of formula I). Includes:

[0051] The present application also provides a method for preparing psilocin (a compound of formula II), comprising:

[0052] [ka]

[0053] Unprotected 4-hydroxyindole (compound of formula III)

[0054] [ka]

[0055] with oxalyl chloride to provide a compound of formula (IV);

[0056] [ka]

[0057] reacting a compound of formula (IV) with dimethylamine (HN(CH3)2) to provide a compound of formula V;

[0058] [ka]

[0059] and The method includes reducing a compound of formula (V) with a reducing reagent to provide psilocin (a compound of formula II).

[0060] Other features and advantages of the present application will become apparent from the following detailed description. However, it should be understood that the detailed description and specific examples, while indicating embodiments of the present application, are given for illustrative purposes only, and that the scope of the claims should not be limited by these embodiments, but should be accorded the broadest interpretation consistent with the description as a whole.

[0061] Embodiments of the present application will now be described in more detail with reference to the accompanying drawings. [Brief explanation of the drawings]

[0062] [Figure 1] FIG. 1 shows the HPLC chromatogram of the compound of formula IV. [Figure 2] FIG. 2 shows the HPLC chromatogram of the compound of formula V. [Figure 3] FIG. 3 shows the 1H-NMR spectrum of the compound of formula V. [Figure 4] FIG. 4 shows the 1H-NMR spectrum of psilocin (compound of formula II). [Figure 5] FIG. 5 shows the 1H-NMR spectrum of chlorodi-tert-butylphosphite. [Figure 6] FIG. 6 is an HPLC chromatogram of psilocybin (compound of formula I). [Figure 7] FIG. 7 is a mass spectrum of psilocybin (compound of formula I). [Figure 8] FIG. 8 shows the 1H-NMR spectrum of psilocybin (compound of formula I). DETAILED DESCRIPTION OF THE INVENTION

[0063] I. Definition Unless otherwise indicated, the definitions and embodiments set forth in this section and other sections are intended to be applicable to all embodiments and aspects of the application where they are appropriate and described herein, as understood by one of ordinary skill in the art.

[0064] As used herein, the term "method of the present application" and like terms refer to methods of preparing psilocybin, psilocin, acid salts of psilocybin, and / or acid salts of psilocin, as described herein.

[0065] As used herein, the term "and / or" means that the listed items are present or used individually or in any combination. In effect, the term means that "at least one of" or "one or more" of the listed items are used or present.

[0066] As used in this application, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, an embodiment including "a solvent" should be understood to present a particular embodiment with one type of solvent, or two or more additional solvents.

[0067] In embodiments that include an "additional" or "second" component, e.g., an additional solvent or second solvent, a second component, as used herein, is chemically distinct from the other or first component. A "third" component is distinct from the other, first, and second components, and further listed or "additional" components are similarly distinct.

[0068] The terms "comprising" (and including any of its forms such as "comprise" and "comprises"), "having" (and including any of its forms such as "have" and "has"), "including" (and including any of its forms such as "include" and "includes"), or "containing" (and including any of its forms such as "contain" and "contains") as used in this application and in the claims are inclusive or open-ended and do not exclude additional, unrecited elements or processes.

[0069] As used herein, the term "consisting of" and its derivatives are intended to be closed-ended terms specifying the presence of stated features, elements, components, groups, integers, and / or steps, and are also intended to exclude the presence of other, unstated features, elements, components, groups, integers, and / or steps.

[0070] As used herein, the term "consisting essentially of" is intended to specify the presence of stated features, elements, components, groups, integers, and / or steps, as well as the presence of features that do not materially affect the basic and novel characteristics of those features, elements, components, groups, integers, and / or steps.

[0071] As used herein, the terms "suitable" and "appropriate" mean that the selection of a particular compound or condition will depend on the specific synthetic operation being performed, the identity of the molecule being converted, and / or the specific use of the compound, but that the selection is well within the skill of one of ordinary skill in the art. All process / method steps described herein should be performed under conditions to provide the indicated product. Those of ordinary skill in the art will understand that, unless otherwise indicated, all reaction conditions, including, for example, reaction solvent, reaction time, reaction temperature, reaction pressure, reactant ratios, and whether the reaction should be performed under an anhydrous or inert atmosphere, can be varied to optimize the yield of the desired product, and that doing so is within the skill of the art.

[0072] As used herein, the terms "about," "substantially," and "approximately" refer to a reasonable amount of deviation from the modified term such that the end result is not significantly altered. These terms of degree should be interpreted as including a deviation of at least ±5% from the modified term if this deviation does not negate the meaning of the word.

[0073] This specification refers to numerous chemical terms and abbreviations used by those of ordinary skill in the art. Nevertheless, definitions of selected terms are provided for clarity and consistency.

[0074] The term "available" as in "available hydrogen atom" or "available atom" refers to an atom that would be known to one of ordinary skill in the art as being capable of being replaced by a substituent.

[0075] As used herein, the term "psilocybin" or "compound of Formula I" or "(I)" has the IUPAC name: 3-[2-(dimethylamino)ethyl]-1H-indol-4-yl dihydrogen phosphate or the chemical name 4-phosphoryloxy-N,N-dimethyltryptamine and has the following chemical formula:

[0076] [ka]

[0077] As used herein, the term "psilocin" or "compound of Formula II" or "(II)" has the IUPAC name: 4-hydroxy-N,N-dimethyltryptamine and has the following chemical formula:

[0078] [ka]

[0079] The term "reducing agent" as used herein means any compound or combination of compounds that reduces a desired functional group. A reducing agent results in the overall addition of electrons, or in the case of organic chemistry, the addition of hydrogen atoms to a functional group.

[0080] As used herein, the term "inert solvent" means a solvent that does not interfere with or otherwise inhibit the reaction. Thus, the identity of the inert solvent will vary depending on the reaction being performed. The selection of an inert solvent is within the skill of one of ordinary skill in the art.

[0081] The term "solvent" includes both a single solvent and a mixture containing two or more solvents.

[0082] As used herein, the term "protecting group" or "PG" and like terms refer to a chemical moiety that protects or masks a reactive portion of a molecule to prevent side reactions at such reactive portion of the molecule while a different portion of the molecule is being manipulated or reacted. After the manipulation or reaction is complete, the protecting group is removed under conditions that do not decompose or degrade other portions of the molecule. The selection of an appropriate protecting group can be made by one skilled in the art. See, for example, "Protective Groups in Organic Chemistry," McOmie, JFW Ed., Plenum Press, 1973, in Greene, TW and Wuts, PGM, "Protective Groups in Organic Synthesis," John Wiley & Sons, 3 rd Many conventional protecting groups are known in the art, as described in Kocienski, P., "Protecting Groups, 3rd Edition," 2003, Georg Thieme Verlag (The Americas), pp. 1999-2003.

[0083] As used herein, the term "Atherton-Todd reaction" refers to the reaction for converting dialkyl phosphites to dialkyl chlorophosphates through the reaction of tetrachloromethane (carbon tetrachloride) in the presence of a base, first described by F.R.A. Atherton, H.T. Openshaw, and A.R.T. Todd in 1945 (Journal of the Chemical Society, pp. 660-663). The base is typically a primary, secondary, or tertiary amine.

[0084] The products of the processes of the present application can be isolated according to known methods, for example, the compounds can be isolated by evaporation of the solvent, filtration, centrifugation, chromatography or other suitable method.

[0085] The term "pharmaceutically acceptable" means compatible with the treatment of a subject.

[0086] The term "pharmaceutically acceptable carrier" refers to a non-toxic solvent, dispersant, excipient, adjuvant or other substance that can be mixed with an active ingredient to produce a pharmaceutical composition, i.e., a dosage form that can be administered to a subject.

[0087] The term "pharmaceutically acceptable salt" means either an acid addition salt or a base addition salt that is suitable or compatible with the treatment of a subject.

[0088] An acid addition salt suitable for or compatible with the treatment of a subject is any non-toxic organic or inorganic acid addition salt of any basic compound.

[0089] As used herein, the term "subject" includes all members of the animal kingdom, including mammals, and suitably refers to humans.

[0090] II. Application Process Applicants have developed a novel, cost-effective, practical, and scalable synthetic route to psilocybin (compound of Formula I), which is 4-phosphoryloxy-N,N-dimethyltryptamine, and its major metabolite, 4-hydroxy-N,N-dimethyltryptamine, psilocin (compound of Formula II). The process of the present application provides a novel method for preparing highly pure psilocybin or psilocin, with a high degree of consistency in purity and yield, and can be easily adopted for commercial production. The prepared psilocybin or psilocin can then be converted to any appropriate pharmaceutically acceptable salt, such as an oxalate, succinate, or benzoate, for dosage form preparation.

[0091] The novel synthetic route to prepare psilocybin and psilocin of the present application has several advantages, including (i) mild reaction conditions, (ii) simple operation, (iii) readily available commercial raw materials, and (iv) suitability for multi-kilogram scale production. An additional advantage is the ability to produce psilocybin and psilocin with improved yield, purity, and reproducibility.

[0092] Starting from unprotected 4-hydroxytryptamine, psilocybin (Formula I) can be synthesized in a five-step procedure via psilocin (Formula II), beginning with the direct condensation of 4-hydroxyindole with oxalyl chloride. Applicants have surprisingly discovered that this condensation step can be accomplished without the need for a protecting group on the OH of 4-hydroxyindole; for example, oxalyl chloride can be directly reacted with unprotected 4-hydroxyindole in an organic solvent such as methyl tert-butyl ether (MTBE) or diethyl ether to yield the di-2-chloro-2-oxoacetyl intermediate (compound of Formula IV). Directly reacting unprotected 4-hydroxyindole with oxalyl chloride is advantageous in that it avoids the 4-hydroxyindole protection and subsequent deprotection steps used in art-known methods for synthesizing psilocin or psilocybin, such as those described above, and further avoids the generation of undesired by-products resulting from additional protection and deprotection steps. Furthermore, the compound of formula IV can be advantageously used in subsequent reaction steps without direct isolation or purification, i.e., the product of the condensation step (compound of formula IV) can be subjected to a subsequent reaction with dimethylamine to provide the oxoacetamide compound of formula V. The compound of formula V can then be reduced, for example with lithium aluminum hydride, to provide psilocin.

[0093] The present applicant has further surprisingly discovered that psilocin can be converted to psilocybin by a novel, optimized phosphorylation reaction using di-tert-butyl phosphite as the phosphorylating reagent to yield the di-tert-butyl phosphite compound of formula VI. For example, psilocin can be phosphorylated with di-tert-butyl phosphite in the presence of a base such as sodium hydroxide or sodium hydroxide in combination with 4-dimethylaminopyridine (DMAP) in an inert solvent such as a mixture of carbon tetrachloride and tetrahydrofuran (THF). Alternatively, psilocin can be phosphorylated with chlorodi-tert-butyl phosphite, which can be generated in situ by combining di-tert-butyl phosphite in the presence of, for example, N-chlorosuccinimide. The latter option has the added advantage of avoiding the use of carbon tetrachloride required in the Atherton-Todd reaction. The resulting di-tert-butyl phosphite compound of formula VI can then be simply hydrolyzed with acid to provide psilocybin. The di-tert-butyl phosphite compound of formula VI can be directly isolated and subsequently incorporated into the subsequent hydrolysis step. Furthermore, the use of di-tert-butyl phosphite as the phosphorylating reagent instead of the O,O-dibenzyl phosphite-type reagents used in the art (see, for example, Hofmann, A. et al., Experientia 1958, 14, 397-399; Helv. Chim. Acta 1959, 42, 2073-2103; and Shirota, O. et al., J. Nat. Prod. 2003, 66, 885-887) is advantageous in that it avoids the formation of the problematic zwitterionic N,O-dibenzyl phosphite derivative by-product (see IIb above) that is generated upon removal of the benzyl group introduced by the O,O-dibenzyl phosphite-type phosphorylating reagent. Alternatively, the di-tert-butyl groups introduced by the di-tert-butyl phosphite reagent as described herein are efficiently removed by acid hydrolysis.

[0094] The applicant, for example, 1 H-NMR, 13 The authenticity of the chemical structures of both psilocybin and its metabolite psilocin was confirmed by C-NMR, IR, HPLC, and / or LCMS.

[0095] The novel synthetic route of the present application has advantages such as improved yield, purity and reproducibility, mild reaction conditions, and environmental friendliness.

[0096] The present application includes a process for preparing psilocybin (compound of formula I), as applicants have discovered that psilocin can be converted to psilocybin via a novel, optimized phosphorylation reaction with di-tert-butyl phosphite.

[0097] [ka]

[0098] The process is: Unprotected psilocin (compound of formula II)

[0099] [ka]

[0100] in the presence of a base (a) di-tert-butyl phosphite

[0101] [ka]

[0102] by reacting with, or Unprotected silosin was treated with (b) chlorodi-tert-butylphosphite [ka]

[0103] to provide a compound of formula (VI);

[0104] [ka]

[0105] and Hydrolyzing the compound of formula VI to provide psilocybin (the compound of formula I). Includes:

[0106] The present application also includes a process for preparing psilocin (compound of formula II), since the applicant has discovered that psilocin can be prepared, for example, by directly reacting unprotected 4-hydroxyindole with oxalyl chloride.

[0107] [ka]

[0108] The process is: Unprotected 4-hydroxyindole (compound of formula III)

[0109] [ka]

[0110] with oxalyl chloride to provide a compound of formula (IV);

[0111] [ka]

[0112] reacting a compound of formula (IV) with dimethylamine (HN(CH3)2) to provide a compound of formula V;

[0113] [ka]

[0114] and The method comprises reducing the compound of formula (V) with a reducing agent to provide psilocin (a compound of formula II).

[0115] Thus, Applicant has discovered that starting from unprotected 4-hydroxytryptamine, psilocin (Formula I) can be synthesized via psilocin in a five-step procedure. Accordingly, the present application includes a process for preparing psilocybin (compound of Formula I).

[0116] [ka]

[0117] The process is: Unprotected 4-hydroxyindole (compound of formula III)

[0118] [ka]

[0119] with oxalyl chloride to provide a compound of formula (IV);

[0120] [ka]

[0121] reacting a compound of formula (IV) with dimethylamine (HN(CH3)2) to provide a compound of formula (V);

[0122] [ka]

[0123] reducing the compound of formula (V) with a reducing reagent to provide psilocin (compound of formula II);

[0124] [ka]

[0125] Silosin was treated with (a) di-tert-butyl phosphite in the presence of a base.

[0126] [ka]

[0127] by reacting with, or Silosin (b) chlorodi-tert-butylphosphite [ka]

[0128] to provide a compound of formula (VI);

[0129] [ka]

[0130] and Hydrolyzing the compound of formula (VI) to provide psilocybin (the compound of formula I). Includes:

[0131] In some embodiments, unprotected 4-hydroxyindole (compound of Formula III) is reacted with an excess of oxalyl chloride to provide a compound of Formula (IV). In some embodiments, unprotected 4-hydroxyindole (compound of Formula III) is reacted with an excess (e.g., about 2 to about 5 molar equivalents, about 2 to about 4 molar equivalents, about 2 to about 3 molar equivalents, about 3 to about 5 molar equivalents, about 3 to about 4 molar equivalents, or about 2.2 molar equivalents) of oxalyl chloride to provide a compound of Formula (IV). In some embodiments, unprotected 4-hydroxyindole (compound of Formula III) is reacted with an excess (e.g., about 2 to about 4 molar equivalents, about 3 molar equivalents, or about 2.2 molar equivalents) of oxalyl chloride to provide a compound of Formula (IV).

[0132] In some embodiments, reacting an unprotected 4-hydroxyindole (a compound of Formula III) with an excess of oxalyl chloride to provide a compound of Formula (IV) comprises adding an unprotected 4-hydroxyindole (a compound of Formula III) to an excess of oxalyl chloride in an inert solvent for a temperature and time suitable for reacting the unprotected 4-hydroxyindole with oxalyl chloride to provide a compound of Formula (IV). In some embodiments, reacting an unprotected 4-hydroxyindole (a compound of Formula III) with an excess of oxalyl chloride to provide a compound of Formula (IV) comprises adding an excess of oxalyl chloride to an unprotected 4-hydroxyindole (a compound of Formula III) in an inert solvent for a temperature and time suitable for reacting the unprotected 4-hydroxyindole with oxalyl chloride to provide a compound of Formula (IV). In some embodiments, the inert solvent is selected from ethers such as diethyl ether, methyl tert-butyl ether (MTBE) and tetrahydrofuran, esters such as ethyl acetate, hydrocarbon solvents such as toluene, and halogenated solvents such as methylene chloride and carbon tetrachloride, and mixtures thereof. In some embodiments, the inert solvent is selected from diethyl ether, methyl t-butyl ether, tetrahydrofuran, ethyl acetate, toluene, methylene chloride, and carbon tetrachloride, and mixtures thereof. In some embodiments, the inert solvent is diethyl ether or MBTE. In some embodiments, the inert solvent is MBTE. In some embodiments, reacting the unprotected 4-hydroxyindole (compound of Formula III) with an excess of oxalyl chloride to provide a compound of Formula (IV) comprises adding the unprotected 4-hydroxyindole (compound of Formula III) to an excess of oxalyl chloride in an inert solvent, such as diethyl ether or MTBE, at a temperature and for a time period for the reaction of the unprotected 4-hydroxyindole with oxalyl chloride to provide a compound of Formula (IV). In some embodiments, the inert solvent is diethyl ether.In some embodiments, reacting the unprotected 4-hydroxyindole (compound of formula III) with an excess of oxalyl chloride to provide the compound of formula (IV) comprises adding an excess of oxalyl chloride to the unprotected 4-hydroxyindole (compound of formula III) in diethyl ether at a temperature and for a time for the reaction of the unprotected 4-hydroxyindole with oxalyl chloride to provide the compound of formula (IV).

[0133] As a representative, non-limiting example, the temperature and time for reacting unprotected 4-hydroxyindole (compound of Formula III) with oxalyl chloride to provide a compound of Formula (IV) are from about 0° C. to about 15° C., from about 0° C. to about 10° C., from about 5° C. to about 10° C., or from about 0° C. to about 5° C., for about 6 hours to about 24 hours, from about 10 hours to about 20 hours, from about 12 hours to about 20 hours, from about 14 hours to about 18 hours, or from about 16 hours. In some embodiments, the temperature and time for reacting unprotected 4-hydroxyindole with oxalyl chloride to provide a compound of Formula (IV) are from about 0° C. to about 10° C., from about 5° C. to about 10° C., or from about 0° C. to about 5° C., for about 12 hours to about 20 hours, from about 14 hours to about 18 hours, or from about 16 hours.

[0134] Thus, in some embodiments, the method comprises reacting unprotected 4-hydroxyindole with about 3 to about 5 molar equivalents, about 3 to about 4 molar equivalents, or about 3 molar equivalents of oxalyl chloride in an inert solvent at about 0° C. to about 5° C. for about 14 to about 18 hours or about 16 hours to provide a compound of Formula (IV). In some embodiments, the method comprises reacting about 3 to about 5 molar equivalents, about 3 to about 4 molar equivalents, or about 3 molar equivalents of oxalyl chloride in diethyl ether at about 0° C. to about 5° C. for about 14 to about 18 hours or about 16 hours to provide a compound of Formula (IV).

[0135] In some embodiments, the method includes reacting unprotected 4-hydroxyindole with, for example, about 2 to about 4 molar equivalents, about 2 to about 3 molar equivalents, or about 2.2 molar equivalents of oxalyl chloride in an inert solvent such as MTBE at a temperature of about 0° C. to about 10° C. or about 5° C. to about 10° C. for about 1 hour to about 4 hours or about 2 hours to about 3 hours to provide a compound of Formula (IV). In some embodiments, the method includes reacting unprotected 4-hydroxyindole with about 2 to about 3 molar equivalents, or about 2.2 molar equivalents of oxalyl chloride in an inert solvent such as MTBE at a temperature of about 5° C. to about 10° C. or about 5° C. for about 2 hours to about 3 hours to provide a compound of Formula (IV).

[0136] In some embodiments, dimethylamine (N(CH3)2) is generated in situ by reacting an acid salt of dimethylamine, such as dimethylamine hydrochloride, with a base. Thus, in some embodiments, a compound of Formula (IV) is reacted with an acid salt of dimethylamine in the presence of a base to provide a compound of Formula (V). In some embodiments, a compound of Formula (V) is prepared by reacting dimethylamine hydrochloride (N(CH3)2·HCl) in the presence of a base to provide a compound of Formula (V). In some embodiments, the base is an organic amine base. In some embodiments, the organic amine base is selected from pyridine, triethylamine, trimethylamine, triphenylamine, tripropylamine, tripentylamine, tert-butylamine, cyclohexylamine, cyclooctylamine, pentylamine, or octylamine. In some embodiments, the organic amine base is triethylamine or pyridine. In some embodiments, the organic amine base is triethylamine. In some embodiments, the organic amine base is pyridine.

[0137] In some embodiments, a compound of Formula (IV) is reacted with an excess amount of a dimethylamine acid salt in the presence of a base to provide a compound of Formula (V). In some embodiments, a compound of Formula (IV) is reacted with about 2 to about 6 molar equivalents, about 3 to about 5 molar equivalents, about 3 molar equivalents, about 4 molar equivalents, about 5 molar equivalents, or about 6 molar equivalents of a dimethylamine acid salt in the presence of a base to provide a compound of Formula (V). In some embodiments, a compound of Formula (IV) is reacted with about 3 to about 5 molar equivalents, about 3 molar equivalents, about 4 molar equivalents, or about 5 molar equivalents of a dimethylamine acid salt in the presence of a base to provide a compound of Formula (V). In some embodiments, a compound of Formula (IV) is reacted with about 3 to about 5 molar equivalents, or about 5 molar equivalents of a dimethylamine acid salt in the presence of a base to provide a compound of Formula (V).

[0138] In some embodiments, a compound of Formula (IV) is reacted with an excess amount of a dimethylamine acid salt in the presence of a base in an inert solvent at a temperature and for a time such that the compound of Formula (IV) reacts with the dimethylamine acid salt in the presence of the base to provide a compound of Formula (V). In some embodiments, the inert solvent is selected from ethers such as diethyl ether and tetrahydrofuran, esters such as ethyl acetate, hydrocarbon solvents such as toluene, and halogenated solvents such as methylene chloride and carbon tetrachloride, and mixtures thereof. In some embodiments, the inert solvent is diethyl ether. In some embodiments, the base is an organic amine base that is liquid at room temperature, such as pyridine. In one embodiment, the base is present in an amount that neutralizes the dimethylamine acid salt, i.e., converts the acid salt to a free base. In some embodiments, the base is present in an excess amount relative to both the compound of Formula (IV) and the dimethylamine acid salt. In some embodiments, the base is used as a solvent or co-solvent.

[0139] As a representative, non-limiting example, the temperature and time for reacting a compound of formula (IV) with a dimethylamine acid salt in the presence of a base to provide a compound of formula (V) are about 18°C to about 25°C, about 20°C to about 25°C, or room temperature, for about 15 minutes to about 2 hours, about 15 minutes to about 1 hour, about 15 minutes to about 45 minutes, about 30 minutes to about 45 minutes, or about 30 minutes.

[0140] Thus, in some embodiments, a compound of formula (IV) is reacted with about 3 to about 5 molar equivalents, or about 5 molar equivalents, of a salt of dimethylamine in the presence of a base at room temperature for about 30 minutes to about 45 minutes, or about 30 minutes, to provide a compound of formula (V).

[0141] In some embodiments, the compound of Formula (IV) that is reacted with an acid salt of dimethylamine in the presence of a base to provide a compound of Formula (V) is crude or not purified, or the compound of Formula (V) is not isolated prior to reacting with an acid salt of dimethylamine in the presence of a base to provide a compound of Formula (V).

[0142] In some embodiments, a compound of formula (IV) is reacted with dimethylamine to provide a compound of formula (V).

[0143] In some embodiments, the compound of Formula (IV) is reacted with an excess of dimethylamine in the presence of a base to provide a compound of Formula (V). In some embodiments, the compound of Formula (IV) is reacted with about 2 to about 6 molar equivalents, about 2 to about 4 molar equivalents, about 2 molar equivalents, about 3 molar equivalents, about 4 molar equivalents, or about 5 molar equivalents, or about 3.2 molar equivalents of dimethylamine in the presence of a base to provide a compound of Formula (V). In some embodiments, the compound of Formula (IV) is reacted with about 3 to about 4 molar equivalents, or about 3.2 molar equivalents of dimethylamine in the presence of a base to provide a compound of Formula (V).

[0144] In some embodiments, a compound of Formula (IV) is reacted with an excess amount of dimethylamine in the presence of an excess amount of base (e.g., about 2 to about 4 molar equivalents, about 2 molar equivalents, about 3 molar equivalents, about 4 molar equivalents, or about 5 molar equivalents, or about 3.2 molar equivalents) at a temperature and for a time period sufficient to react the compound of Formula (IV) with the dimethylamine to provide a compound of Formula (V). In some embodiments, the base is an organic amine base that is liquid at room temperature, such as triethylamine or pyridine. In some embodiments, the base is used as a solvent or cosolvent. In some embodiments, the base is triethylamine (TEA). In some embodiments, the base is TEA, and the TEA is used as a solvent or cosolvent. In some embodiments, the dimethylamine and base are used in equal amounts.

[0145] Thus, in some embodiments, a compound of formula (IV) is reacted with an excess (e.g., 3 to about 4 molar equivalents, or about 3.2 molar equivalents) of dimethylamine in the presence of an excess (e.g., about 3 to about 4 molar equivalents, or about 3.2 molar equivalents) of triethylamine to provide a compound of formula (V).

[0146] In some embodiments, a compound of Formula (IV) is reacted with an excess of dimethylamine in the presence of an excess of base, such as TEA, to provide a compound of Formula (V) by combining dimethylamine with a base to form a solution of dimethylamine in the base, adding the solution to a compound of Formula (IV) at a temperature of about 0°C to about 15°C, about 0°C to about 10°C, or about 5°C to about 10°C, or about 5°C to form a reaction mixture, and then warming the reaction mixture to about 18°C to about 25°C, about 20°C to about 25°C, or room temperature, and stirring the reaction mixture for about 2 hours to about 5 hours, about 3 hours to about 5 hours, about 3 hours to about 4 hours, or about 3 hours. Thus, in some embodiments, a compound of Formula (IV) is reacted with an excess of dimethylamine in the presence of an excess of base, such as TEA, at a temperature of about 0°C to about 10°C, or about 5°C to about 10°C to form a reaction mixture, warming the reaction mixture to room temperature, and stirring the reaction mixture for about 3 hours to about 5 hours to provide a compound of Formula (V).

[0147] In some embodiments, the compound of formula (IV) that is reacted with dimethylamine to provide the compound of formula (V) is crude or unpurified, or the compound of formula (IV) is not isolated prior to reacting with dimethylamine to provide the compound of formula (V).

[0148] In some embodiments, the reducing agent that provides psilocin (II) from the compound of formula (V) is any suitable reducing agent that reduces the ketone group of the compound of formula (V) to an alkane.

[0149] In some embodiments, the compound of formula (V) is reduced with a reducing agent to provide psilocin (a compound of formula II) using any suitable conditions known in the art for reducing a compound of formula (V) with a reducing agent to provide psilocin.

[0150] In some embodiments, the compound of Formula (V) is reduced with an excess (e.g., about 1.5 to about 3 molar equivalents, or about 1.5 to about 2 molar equivalents, or about 2 molar equivalents) of reducing agent in an inert solvent at a temperature and for a time such that the compound of Formula (V) is reduced with the reducing agent to provide silosin. In some embodiments, the inert solvent is selected from ethers, such as diethyl ether and tetrahydrofuran, hydrocarbon solvents, such as toluene, and mixtures thereof. In some embodiments, the inert solvent is selected from diethyl ether and toluene. In some embodiments, the inert solvent is tetrahydrofuran. As representative, non-limiting examples of temperatures and reaction times, the reaction is heated to boiling (reflux) and maintained at boiling for about 1 hour to about 6 hours, about 2 hours to about 6 hours, about 3 hours to about 5 hours, about 4 hours to about 5 hours, or about 4 hours.

[0151] In some embodiments, a suitable reducing agent for reducing the compound of Formula (V) is a metal hydride. In some embodiments, the metal hydride is selected from lithium borohydride, sodium borohydride, and lithium aluminum hydride. In some embodiments, the metal hydride is lithium aluminum hydride. In some embodiments, the lithium aluminum hydride is provided as a solution in an inert solvent. Thus, in some embodiments, the lithium aluminum hydride is a lithium aluminum hydride solution. In some embodiments, the lithium aluminum hydride is a lithium aluminum hydride solution in THF. In some embodiments, the lithium aluminum hydride is a 1.0 M lithium aluminum hydride solution in THF.

[0152] Thus, in some embodiments, the compound of formula (V) is reacted with about 1.5 to about 2 molar equivalents, or about 2 molar equivalents, of lithium aluminum hydride in THF at reflux temperature for about 3 hours to about 5 hours, about 4 hours to about 5 hours, or about 4 hours to provide psilocin (II).

[0153] In some embodiments, silosin is reacted with di-tert-butyl phosphite in the presence of a base under modified Atherton-Todd reaction conditions to provide a compound of Formula (VI). In some embodiments, silosin is reacted with a slight excess (e.g., about 1.1 to about 1.5 molar equivalents) of di-tert-butyl phosphite in the presence of an excess of base (e.g., about 1.5 to about 2.5 molar equivalents or about 2 molar equivalents) in an inert solvent at a temperature and for a time sufficient for the silosin to react with the di-tert-butyl phosphite to provide a compound of Formula (VI). In some embodiments, the inert solvent is selected from ethers such as diethyl ether and tetrahydrofuran, esters such as ethyl acetate, hydrocarbon solvents such as toluene, halogenated solvents such as methylene chloride and carbon tetrachloride, and mixtures thereof. In some embodiments, the inert solvent is selected from tetrahydrofuran and carbon tetrachloride, and mixtures thereof. In some embodiments, the inert solvent is a mixture of tetrahydrofuran and carbon tetrachloride. In some embodiments, the inert solvent is a 1:1 v / v mixture of tetrahydrofuran and carbon tetrachloride. In some embodiments, the base is a weak base. In some embodiments, the base is selected from (1,4-diazabicyclo[2.2.2]octane) (DABCO); 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU); 4-dimethylaminopyridine (DMAP); alkylamines such as triethylamine and diethylamine; carbonates such as sodium carbonate and potassium carbonate; bicarbonates such as sodium bicarbonate and potassium bicarbonate; and hydroxides such as sodium hydroxide and potassium hydroxide, and combinations thereof. In some embodiments, the base is sodium hydroxide. In some embodiments, the base is sodium hydroxide in combination with DMAP. In some embodiments, representative non-limiting examples of temperatures and reaction times include reacting silosin with di-tert-butyl phosphite at about 18°C to about 50°C, about 20°C to about 5°C, or room temperature for about 12 hours to about 20 hours, about 14 hours to about 18 hours, or about 16 hours.

[0154] Thus, in some embodiments, silosin is reacted with a slight excess (e.g., about 1.1 to about 1.5 molar equivalents) of di-tert-butyl phosphite in a mixture of tetrahydrofuran and carbon tetrachloride in the presence of an excess (e.g., about 2 molar equivalents) of sodium hydroxide or a combination of about 0.1 molar equivalents of 4-dimethylaminopyridine (DMAP) in combination with an excess (e.g., about 2 molar equivalents) of sodium hydroxide at room temperature for about 12 hours to about 20 hours, about 14 hours to about 18 hours, or about 16 hours to provide a compound of formula VI.

[0155] In some embodiments, silosin is reacted with chlorodi-tert-butylphosphite to provide a compound of formula (VI). In some embodiments, chlorodi-tert-butylphosphite is prepared in situ by reacting di-tert-butylphosphite with N-chlorosuccinimide (NCS) to provide chlorodi-tert-butylphosphite. Thus, in some embodiments, the process comprises reacting silosin with di-tert-butylphosphite and N-chlorosuccinimide (NCS) to provide a compound of formula (VI). Thus, in some embodiments, the step of reacting silosin with di-tert-butylphosphite and N-chlorosuccinimide to provide a compound of formula (VI) comprises: reacting di-tert-butyl phosphite with N-chlorosuccinimide (NCS) to provide chlorodi-tert-butyl phosphite; and reacting silosin with chlorodi-tert-butylphosphite to provide a compound of formula (VI); Includes:

[0156] In some embodiments, di-tert-butyl phosphite is reacted with a slight excess (e.g., 1.1 to about 2 molar equivalents, or about 1.2 molar equivalents) of NCS in an inert solvent such as THF at a temperature and for a time to provide chlorodi-tert-butyl phosphite. As a representative, non-limiting example of temperature and reaction time, in some embodiments, tert-butyl phosphite is reacted with a slight excess (e.g., about 1.1 to about 2 molar equivalents, or about 1.2 molar equivalents) of NCS at about 0°C to about 15°C, about 5°C to about 10°C, or about 10°C for about 1 hour to about 4 hours, about 2 hours to about 4 hours, about 2 hours to about 3 hours, or about 2 hours.

[0157] In some embodiments, reacting silosin with chlorodi-tert-butylphosphite to provide a compound of Formula (VI) comprises combining silosin with a slight excess (e.g., about 1.1 to about 2 molar equivalents, or about 1.1 to about 1.5 molar equivalents) of chlorodi-tert-butylphosphite at about 0° C. to about 15° C., or about 5° C. to about 10° C., or 5° C., for about 2 hours to about 8 hours, about 2 hours to about 6 hours, or about 3 hours to about 6 hours to provide a compound of Formula (VI).

[0158] In some embodiments, psilocybin (I) is prepared using any suitable conditions known in the art for hydrolyzing a compound of formula (VI) to provide psilocybin.

[0159] In some embodiments, the compound of Formula (VI) is hydrolyzed using an acid to provide psilocybin. Thus, in some embodiments, the process includes hydrolyzing the compound of Formula (VI) with an acid to provide psilocybin. In some embodiments, the acid is hydrochloric acid. In some embodiments, the compound of Formula (VI) is reacted with an excess amount of acid (e.g., about 1.1 to about 1.5 molar equivalents) in a suitable solvent at a temperature and for a time to provide psilocybin. As representative, non-limiting examples, the temperature and time for hydrolyzing the compound of Formula (VI) with an acid to provide psilocybin are about 18°C to about 25°C, about 20°C to about 25°C, or room temperature, for about 1 hour to about 4 hours, about 1 hour to about 3 hours, about 1.5 hours to about 2.5 hours, or about 2 hours.

[0160] Thus, in some embodiments, the compound of formula (VI) is hydrolyzed with hydrochloric acid in acetone at room temperature for about 1 hour to about 3 hours or about 2 hours to provide psilocybin.

[0161] In some embodiments, the compound of formula (VI) that is hydrolyzed to provide psilocybin is crude or unpurified, or the compound of formula (IV) is not isolated prior to hydrolysis to provide psilocybin.

[0162] In some embodiments, the compound of formula (VI) is hydrolyzed with an acid in a suitable solvent at a temperature and for a time to form a reaction mixture containing psilocybin, and the psilocybin is isolated from the reaction mixture. Thus, in some embodiments, the process comprises hydrolyzing the compound of formula (VI) to provide a reaction mixture containing psilocybin, and isolating the psilocybin.

[0163] In some embodiments, psilocybin is isolated from the reaction mixture using any suitable conditions known in the art for isolating a product, such as psilocybin, from a reaction mixture.

[0164] In some embodiments, the step of isolating psilocybin comprises: Adjusting the pH of the reaction mixture to pH 6-7; Extracting impurities from the reaction mixture using a suitable first solvent: extracting the psilocybin from the reaction mixture with a suitable second solvent to provide a psilocybin solution; Concentrating the psilocybin solution and crystallizing psilocybin from the psilocybin solution; and Separating psilocybin from a crude psilocybin solution to provide psilocybin. Includes:

[0165] In some embodiments, the pH of the reaction mixture is adjusted using a base.

[0166] In some embodiments, a suitable first solvent is isopropyl alcohol, 2-methyltetrahydrofuran, or water, a mixture thereof.

[0167] In some embodiments, a suitable second solvent is water or heptane.

[0168] In some embodiments, the first solvent is isopropyl alcohol and the second solvent is water. In some embodiments, the first solvent is 2-methyltetrahydrofuran and the second solvent is heptane. In some embodiments, the first solvent is water and the second solvent is heptane.

[0169] In some embodiments, the step of concentrating the psilocybin solution is by distillation or rotary evaporation.

[0170] In some embodiments, the separation of psilocybin from the psilocybin solution is by filtration. In some embodiments, the separation of psilocybin from the psilocybin solution is by filtration using a filter aid. In some embodiments, the filter aid is diatomaceous earth (DE), perlite, cellulose, and combinations thereof.

[0171] In some embodiments, the process further comprises purifying the psilocybin, hi some embodiments, the psilocybin is purified using any suitable conditions known in the art for purifying products such as psilocybin.

[0172] In some embodiments, purifying psilocybin comprises: dissolving psilocybin in water to provide an aqueous psilocybin solution; Neutralizing the aqueous psilocybin solution; extracting psilocybin from the aqueous psilocybin solution using a suitable solvent to form a pure psilocybin solution; Concentrating the pure psilocybin solution and crystallizing pure psilocybin from the pure psilocybin solution; and Separating pure psilocybin from a pure psilocybin solution to provide pure psilocybin Includes:

[0173] In some embodiments, the neutralization of the aqueous psilocybin solution is with an acid.

[0174] In some embodiments, the step of concentrating the pure psilocybin solution is by distillation or rotary evaporation.

[0175] In some embodiments, separating the pure psilocybin from the pure psilocybin solution is by filtration.

[0176] In some embodiments, the processes of the present application provide intermediates comprising a desired polymorphic form of psilocin.

[0177] In some embodiments, crystallizing pure psilocybin provides a desired polymorphic form of psilocybin.

[0178] In some embodiments, crystallizing pure psilocybin provides pure psilocybin in a consistent polymorphic form for administration to a human subject.

[0179] In some embodiments, the methods provide intermediates, including but not limited to psilocin, various polymorphic forms of psilocybin, and prodrugs and analogs thereof, and formulations thereof for use in medicine.

[0180] In an exemplary embodiment of the process of the present application, the present application includes a process for preparing psilocybin (the compound of Formula I).

[0181] [ka]

[0182] The process is: Unprotected 4-hydroxyindole (compound of formula III)

[0183] [ka]

[0184] with an excess of oxalyl chloride to provide a compound of formula (IV);

[0185] [ka]

[0186] reacting a compound of formula (IV) with an excess of dimethylamine hydrochloride (HN(CH3)2·HCl) in the presence of a base to provide a compound of formula (V);

[0187] [ka]

[0188] reducing the compound of formula (V) with a reducing reagent to provide psilocin (compound of formula II);

[0189] [ka]

[0190] Silosin and di-tert-butyl phosphite

[0191] [ka]

[0192] in the presence of a base to provide a compound of formula VI;

[0193] [ka]

[0194] and Hydrolyzing the compound of formula VI to provide psilocybin (the compound of formula I). Includes:

[0195] In some embodiments, unprotected 4-hydroxyindole (compound of formula III) is reacted with oxalyl chloride in diethyl ether to provide a compound of formula (IV).

[0196] In some embodiments, unprotected 4-hydroxyindole (compound of formula III) is reacted with oxalyl chloride in methyl tert-butyl ether (MTBE) to provide a compound of formula (IV).

[0197] In some embodiments, the compound of formula IV is not isolated prior to reaction with excess dimethylamine hydrochloride in the presence of a base.

[0198] In some embodiments, the compound of formula (IV) is reacted with an excess of dimethylamine hydrochloride (HN(CH3)2·HCl) in the presence of pyridine to provide the compound of formula VI.

[0199] In some embodiments, silosin is reacted with di-tert-butyl phosphite in a mixture of tetrahydrofuran and carbon tetrachloride in the presence of sodium hydroxide, or a combination of sodium hydroxide in combination with 4-dimethylaminopyridine (DMAP) to provide a compound of formula (VI).

[0200] In some embodiments, the compound of formula VI is not isolated prior to hydrolysis with acid to provide psilocybin.

[0201] In an exemplary embodiment of the process of the present application, the present application includes a process for preparing psilocybin (the compound of Formula I).

[0202] [ka]

[0203] The process is: Unprotected 4-hydroxyindole (compound of formula III)

[0204] [ka]

[0205] with an excess of oxalyl chloride to provide a compound of formula (IV);

[0206] [ka]

[0207] reacting said compound of formula (IV) with an excess of dimethylamine to provide a compound of formula (V);

[0208] [ka]

[0209] reducing the compound of formula (V) with a reducing reagent to provide psilocin (compound of formula II);

[0210] [ka]

[0211] Silosin with chlorodi-tert-butylphosphite

[0212] [ka]

[0213] to provide a compound of formula VI;

[0214] [ka]

[0215] and hydrolyzing the compound of formula VI to provide psilocybin (the compound of formula I). Includes:

[0216] In some embodiments, chlorodi-tert-butylphosphite is prepared in situ by reacting di-tert-butylphosphite with N-chlorosuccinimide (NCS) to provide chlorodi-tert-butylphosphite. Thus, in some embodiments, the process comprises reacting psilocin with di-tert-butylphosphite and N-chlorosuccinimide (NCS) to provide a compound of formula (VI). Thus, in an exemplary embodiment of the process of the present application, the application includes a process for preparing psilocybin (a compound of formula I).

[0217] [ka]

[0218] The process is: Unprotected 4-hydroxyindole (compound of formula III)

[0219] [ka]

[0220] with an excess of oxalyl chloride to provide a compound of formula (IV);

[0221] [ka]

[0222] reacting said compound of formula (IV) with an excess of dimethylamine to provide a compound of formula (V);

[0223] [ka]

[0224] reducing the compound of formula (V) with a reducing reagent to provide psilocin (compound of formula II);

[0225] [ka]

[0226] Silosin and di-tert-butyl phosphite

[0227] [ka]

[0228] and N-chlorosuccinimide (NCS) to provide a compound of formula VI;

[0229] [ka]

[0230] and hydrolyzing the compound of formula VI to provide psilocybin (the compound of formula I). Includes:

[0231] In some embodiments, unprotected 4-hydroxyindole (compound of formula III) is reacted with oxalyl chloride in methyl tert-butyl ether (MTBE) to provide a compound of formula (IV).

[0232] In some embodiments, the compound of formula IV is not isolated prior to reaction with an excess of dimethylamine.

[0233] In some embodiments, a compound of formula (IV) is reacted with an excess of dimethylamine in the presence of a base to provide a compound of formula VI. In some embodiments, a compound of formula (IV) is reacted with an excess of dimethylamine in the presence of triethylamine to provide a compound of formula VI. In some embodiments, triethylamine is a solvent.

[0234] In some embodiments, the compound of formula VI is not isolated prior to hydrolysis with acid to provide psilocybin.

[0235] In an exemplary embodiment of the process of the present application, the present application includes a process for preparing each of psilocybin of formula (I) and its active metabolite, psilocin of formula (II).

[0236] [ka]

[0237] The process is: Unprotected 4-hydroxyindole of formula III

[0238] [ka]

[0239] with about 3 equivalents of oxalyl chloride at about 0° C. for about 16 hours to provide a compound of formula (IV);

[0240] [ka]

[0241] reacting the crude compound of formula (IV) with excess dimethylamine hydrochloride in diethyl ether in the presence of pyridine at room temperature for about 30 minutes to provide a compound of formula (V);

[0242] [ka]

[0243] reducing the compound of formula (V) with excess lithium aluminum hydride in a 1.0 M solution of pure psilocybin in tetrahydrofuran (THF) at reflux for about 4 hours to provide a compound which is psilocin of formula (II); Reacting silosin with about 1.1 equivalents of di-tert-butyl phosphite in carbon tetrachloride (CCl4) and tetrahydrofuran (THF) in a ratio of CCl4 to THF of 1:1 (v / v) in the presence of a weak base at about room temperature for about 6 hours to provide a di-tert-butyl compound of formula (VI):

[0244] [ka]

[0245] hydrolyzing the compound of formula (VI) with about 6 M hydrogen chloride in acetone to form a reaction mixture, and stirring the reaction mixture at room temperature for about 2 hours; Neutralize the reaction mixture with a base to a pH of 6-7; extracting the liberated impurities from the reaction mixture with a first suitable organic solvent; extracting the crude psilocybin from the reaction mixture with a second suitable organic solvent to form a psilocybin solution; concentrating the psilocybin solution to crystallize crude psilocybin; isolating crude psilocybin by filtration; dissolving crude psilocybin in water to form an aqueous psilocybin solution, and neutralizing the aqueous psilocybin solution with a weak acid; extracting pure psilocybin base from the aqueous psilocybin solution with a suitable solvent to form a pure psilocybin solution; concentrating the pure psilocybin solution to crystallize pure psilocybin from the pure psilocybin solution; and and isolating the pure psilocybin from the pure psilocybin solution by filtration.

[0246] In some embodiments, the present application also includes novel process synthetic routes for preparing each of psilocybin of formula (I) and its active metabolite, psilocin of formula (II).

[0247] [ka]

[0248] The process is: (i) reacting an unprotected 4-hydroxyindole of formula (III);

[0249] [ka]

[0250] (ii) adding about 3 equivalents of oxalyl chloride to a solution of unprotected 4-hydroxyindole in an organic solvent at a temperature of about 0° C. for about 16 hours to obtain 3-(2-chloro-2-oxoacetyl)-1H-indol-4-yl 2-chloro-2-oxoacetate of formula (IV);

[0251] [ka]

[0252] (iii) adding excess dimethylamine hydrochloride to a diethyl ether / pyridine solution of crude 3-(2-chloro-2-oxoacetyl)-1H-indol-4-yl 2-chloro-2-oxoacetate of formula (IV) at room temperature for about 30 minutes to provide exclusively 2-(4-hydroxy-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide of formula V;

[0253] [ka]

[0254] (iv) adding a 1.0 M solution of lithium aluminum hydride in excess tetrahydrofuran (THF) to 2-(4-hydroxy-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide of formula V in refluxing THF for about 4 hours to provide 3-(2-(dimethylamino)ethyl)-1H-indol-4-ol (psilocin) of formula II; Adding 1.1 equivalents of di-tert-butyl phosphite to a weak base in a solution of carbon tetrachloride (CCl4) and tetrahydrofuran (THF) in a CCl4 to THF ratio of 1:1 (v / v) at room temperature for about 6 hours to obtain crude di-tert-butyl(3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate of formula (VI);

[0255] [ka]

[0256] (vii) adding about 6 M hydrogen chloride in acetone to crude di-tert-butyl(3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate; (viii) stirring the reaction mass at room temperature and maintaining the reaction mass at room temperature for 2 hours; (ix) Neutralize with a base to pH 6-7; (x) removing liberated impurities from the reaction mixture by extraction into an organic solvent; (xi) extracting the crude psilocybin base into an organic solvent; (xii) distilling the solvent to form a crude psilocybin solution in the organic solvent; (xiii) isolating the crude psilocybin solution by filtration; (xiv) dissolving crude psilocybin in an aqueous medium and neutralizing with a weak acid; (xv) extracting pure psilocybin base into a solvent; (xvi) partial distillation of the solvent and crystallization of psilocybin base from the same solvent; and (xvii) isolating the pure psilocybin base by filtration.

[0257] In some embodiments, the reaction temperature in step (i) is 0 to 100°C, more preferably 0 to 5°C.

[0258] In some embodiments, the organic solvent in step (i) is diethyl ether. In some embodiments, the organic solvent in step (i) is methyl tert-butyl ether.

[0259] In some embodiments, the excess dimethylamine hydrochloride used in step (ii) is 3 equivalents, preferably 5 equivalents.

[0260] In some embodiments, the organic solvent used in step (i), (ii), or (iii) is selected from an inert solvent such as tetrahydrofuran, a hydrocarbon solvent such as toluene, and preferably toluene.

[0261] In some embodiments, the excess lithium aluminum used in step (iii) is 2 equivalents.

[0262] In some embodiments, the weak base used in step (iv) is selected from (1,4-diazabicyclo[2.2.2]octane) (DABCO), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), or alkylamines such as triethylamine and diethylamine, carbonates or bicarbonates, preferably sodium or potassium carbonate or sodium or potassium hydroxide, preferably sodium hydroxide, preferably sodium hydroxide.

[0263] In some embodiments, the organic solvent used in the hydrolysis in step (iv) is selected from ketones, preferably acetone.

[0264] In some embodiments, the solvent used in the purification in step (iv) is selected from alcohols such as methanol, ethanol, isopropanol, ketones such as acetone, methyl ethyl nitriles such as acetonitrile, in combination with or without water.

[0265] In some embodiments, purification provides a method for crystallizing a desired polymorphic form of psilocybin.

[0266] In some embodiments, crystallization provides chemically pure psilocybin of consistent polymorphic form for administration to human subjects.

[0267] In some embodiments, the crude psilocybin obtained by the methods of the present application starting with 4-hydroxyindole (e.g., psilocybin prior to purification steps) has a purity of greater than about 95%, wherein any one impurity is less than about 1.5%. In some embodiments, the crude psilocybin obtained by the processes of the present application (e.g., psilocybin prior to purification steps) has a purity of greater than about 90%, greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, or greater than about 98%, wherein any one impurity is less than about 1.5%.

[0268] In some embodiments, the crude psilocybin obtained by the present methods contains less than about 5% psilocin, less than about 4% psilocin, less than about 3% psilocin, less than about 2% psilocin, or less than about 1% psilocin.

[0269] In some embodiments, the purity of pure psilocybin obtained by the methods of the present application starting from 4-hydroxyindole is greater than about 98.0%, where any one impurity is less than about 0.2%, or about 0.15%, and the purity of psilocybin is greater than about 98.0%, or greater than about 99%, with total impurities being less than about 0.15%, or about 0.10%.

[0270] In some embodiments, the pure psilocybin obtained by the present methods contains less than about 1% psilocin.

[0271] In some embodiments, the present application also includes HPLC methods for the analysis and assay of psilocybin, psilocin, and intermediate compounds of formulas (III), (IV), (V), and (VI). In some embodiments, the present application includes an HPLC method for assaying psilocybin, comprising: Dissolve the psilocybin sample in a dilution of 0.1% H3PO4 in acetonitrile:water (1:1); Inject the sample solution (approximately 10 μl) into a 100 mm × 4 mm, 3 μm RP-18 HPLC column; Eluting the sample from the column at 1 ml / min using a mixture of acetonitrile (28 vol%) and ammonium format buffer (72 vol%, 0.005 M, pH-4) as the eluent; and Measure the psilocybin content of relevant samples using a UV detector at a wavelength of 245 nm.

[0272] Those skilled in the art will appreciate that further manipulation of substituents on the intermediate and final compounds in the above schemes can be carried out using known chemistry to provide alternative compounds of the present application.

[0273] Psilocybin and psilocin can be used in the form of pharmaceutically acceptable salts. Those skilled in the art will recognize examples of salts that the psilocybin and psilocin of the present invention can form. Examples of such psilocybin and psilocin compounds are described herein with reference to possible salts. Such references are for illustrative purposes only. Pharmaceutically acceptable salts can be used with psilocybin and psilocin to treat subjects. However, non-pharmaceutical salts may be useful in preparing psilocybin and psilocin intermediates. The term "pharmaceutically acceptable salt" refers to salts (including internal salts such as zwitterions) that have the same efficacy as the parent compound and are not biologically or otherwise undesirable (e.g., not toxic or harmful to the recipient). Thus, embodiments of the present invention provide pharmaceutically acceptable salts of psilocybin and / or psilocin. As used herein, the term "salt" refers to both acidic salts formed with inorganic and / or organic acids and basic salts formed with inorganic and / or organic bases.

[0274] Salts of psilocybin and / or psilocin can be formed by methods known to those skilled in the art, such as by reacting psilocybin and / or psilocin with an amount, e.g., an equal amount, of an acid or base in a medium such that the salt precipitates or in an aqueous medium, followed by lyophilization.

[0275] In some embodiments, the pharmaceutically acceptable salt is an acid addition salt or a base addition salt. The selection of an appropriate salt will be made by one skilled in the art (see, for example, SM Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19).

[0276] Acid addition salts suitable for or compatible with the treatment of subjects include any non-toxic organic or inorganic acid addition salts of any basic compound. Because both compounds (I) and (II) contain amine groups, acid addition salts can be formed. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, as well as acid metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include mono-, di-, and tri-carboxylic acids. Examples of such organic acids include acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, mandelic acid, salicylic acid, 2-phenoxybenzoic acid, p-toluenesulfonic acid, and other sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, and 2-hydroxyethanesulfonic acid. In some embodiments, mono- or di-acid salts are formed, and such salts exist in hydrated, solvated, or substantially anhydrous forms. In general, acid addition salts are more soluble in water and various hydrophilic organic solvents and generally exhibit higher melting points than their free base forms. Criteria for selecting an appropriate salt are known to those skilled in the art. Other pharmaceutically unacceptable salts, such as, but not limited to, oxalates, can be used, for example, in isolating the compounds of the present application for laboratory use, or for subsequent conversion to pharmaceutically acceptable acid addition salts. Exemplary acid addition salts include acetate, ascorbate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphorate, camphorsulfonate, fumarate, hydrochloride, hydrobromide, hydroiodide, lactate, maleate, methanesulfonate ("mesylate"), naphthalenesulfonate, nitrate, oxalate, phosphate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, and toluenesulfonate (also known as tosylate).Suitable salts include, for example, acid addition salts that can be formed by mixing a solution of compound with a solution of pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, acetic acid, trifluoroacetic acid or benzoic acid.In addition, acids that are generally considered suitable for forming pharmaceutically useful salts from basic pharmaceutical compounds are discussed in, for example, P. Stahl et al., Camille G. (eds.) and Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley VCH; S. Berge et al., Journal of Pharmaceutical Sciences 1977 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and The Orange Book (Food & Drug Administration, Washington, DC on their website).These disclosures are incorporated herein by reference.

[0277] All such acid salts are intended to be pharmaceutically acceptable salts within the scope of the present invention, and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the present invention. Furthermore, it is understood that the psilocybin and psilocin of the present invention contain both acidic and basic moieties, and thus the psilocybin of the present invention may exist in a zwitterionic form, possessing both an anionic and a cationic center within the same psilocybin, resulting in a net neutral charge. Such zwitterions are encompassed by the present invention.

[0278] The formation of solvates varies depending on the compound and the solvate. Generally, solvates are formed by dissolving the compound in an appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. Selection of appropriate conditions for forming a particular solvate can be performed by one of ordinary skill in the art. Examples of suitable solvents are ethanol, water, etc. When water is the solvent, the molecule is called a "hydrate." The formation of solvates of the compounds of the present application varies depending on the compound and the solvate. Generally, solvates are formed by dissolving the compound in an appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. Selection of appropriate conditions for forming a particular solvate can be performed by one of ordinary skill in the art.

[0279] Isotopically enriched compounds of the present application and pharmaceutically acceptable salts, solvates and / or prodrugs thereof can be prepared without undue experimentation by conventional techniques well known to those skilled in the art, or by processes analogous to those described in the schemes and examples herein, using appropriate isotopically enriched reagents and / or intermediates.

[0280] It should be understood that throughout the processes described herein, suitable protecting groups are added to, and subsequently removed from, various reactants and intermediates, where appropriate, in a manner readily understood by one of ordinary skill in the art. Conventional procedures for using such protecting groups, as well as examples of suitable protecting groups, are described, for example, in "Protective Groups in Organic Synthesis" (TW Green, PGM Wuts, Wiley-Interscience, New York, (1999)). It should also be understood that any intermediate or final product on the synthetic route toward the final product can be subjected to conversion of a group or substituent to another group or substituent by chemical manipulation, where the types of conversion possible are limited only by the inherent incompatibility of other functional groups carried by the molecule at that stage with the conditions or reagents used in the conversion. Such inherent incompatibilities and ways to circumvent them by carrying out the appropriate conversions and synthetic steps in the appropriate order will be readily apparent to one of ordinary skill in the art. While exemplary conversions are presented herein, it should be understood that the described conversions are not limited to only the generic groups or substituents for which the conversions are exemplified. References and descriptions of other suitable transformations can be found in "Comprehensive Organic Transformations - A Guide to Functional Group Preparations" (RC Larock, VHC Publishers, Inc. (1989)). References and descriptions of other suitable reactions can be found in organic chemistry textbooks, such as "Advanced Organic Chemistry" (March, 4th ed. McGraw Hill (1992)) or "Organic Synthesis" (Smith, McGraw Hill, (1994)). Techniques for purification of intermediates and final products include, for example, straight and reversed phase chromatography on columns or spinning plates, recrystallization, distillation, and liquid-liquid or solid-liquid extraction, and will be readily apparent to those skilled in the art.

[0281] III. Compounds and Compositions The present application provides methods for preparing psilocybin and psilocin. Accordingly, the present application includes psilocybin prepared by the process of the present application described above. The present application also includes psilocin prepared by the process of the present application described above.

[0282] The present application also includes compounds of formula IV.

[0283] [ka]

[0284] The present application also includes compounds of formula VI.

[0285] [ka]

[0286] In some embodiments, psilocybin, psilocin, and intermediates thereof, or pharmaceutically acceptable salts thereof, are exemplified below.

[0287] [ka]

[0288] Psilocybin and psilocin can be used in the form of pharmaceutically acceptable salts as described above.

[0289] And / or psilocin may further exist in various polymorphic forms, and any polymorphs or mixtures thereof are contemplated to be included within the scope of this application.

[0290] Psilocybin and / or psilocin may further be radiolabeled, and therefore all radiolabeled versions of psilocybin and / or psilocin are included within the scope of this application. Psilocybin and / or psilocin also include those having one or more radioactive atoms incorporated within their structure.

[0291] The psilocybin and / or psilocin of the present application are suitably formulated into compositions using one or more carriers in a conventional manner. The psilocybin and psilocin of the present application are suitably formulated into pharmaceutical compositions for administration to a subject in a biologically compatible form suitable for in vivo administration. In embodiments of the present application, the pharmaceutical compositions are used to treat any disease, disorder, or condition, such as mental health disorders, including major depressive disorder, anxiety, and addictive disorders, among other CNS disorders. [Example]

[0292] The following non-limiting examples are illustrative of the present application.

[0293] General Method All starting materials used herein are either commercially available or have been previously described in the literature. 1 H-NMR spectrum and 13 The C-NMR spectrum 1 H-NMR was recorded on either a Bruker 300, Bruker DPX400, or Varian +400 spectrometer operating at 300, 400, and 400 MHz, respectively, in deuterated chloroform as solvent, using TMS or the residual solvent signal as internal reference, unless otherwise indicated. All reported chemical shifts are in ppm on the delta scale, and fine divisions of signals as they appear in the recordings are generally indicated, for example, as follows: s: singlet, br s: broad singlet, d: doublet, t: triplet, q: quartet, m: multiplet. Unless otherwise indicated, in the following tables: 1 H-NMR data was obtained at 400 MHz using CDCl3 as the solvent.

[0294] Product purification was performed using Chem Elut Extraction Columns (Varian, Cat. No. 1219-8002), Mega BE-SI (Bond Elut Silica) SPE Columns (Varian, Cat. Nos. 12256018; 12256026; 12256034), or by flash chromatography in silica-packed glass columns.

[0295] Example 1: Exemplary Preparation of Psilocin (II) and Psilocybin (I)

[0296] [ka]

[0297] Step 1: 3-(2-chloro-2-oxoacetyl)-1H-indol-4-yl 2-chloro-2-oxoacetate (compound of formula IV) Oxalyl chloride (2.1 equiv.) was dissolved in 2 V of methyl tert-butyl ether (MTBE) and cooled to 5°C. Unprotected 4-hydroxyindole (1 equiv.) was dissolved in 8 V of MTBE and slowly added to the cooled oxalyl chloride solution via a dropwise addition column. The reaction mixture was stirred for 2-3 h. An in-process control (IPC) sample was taken by withdrawing 0.05 ml and diluting it in 20 ml of HPLC-grade acetonitrile. The IPC passed with less than 5% 4-hydroxyindole (HPLC, see Figure 1). The reaction mixture was loaded onto Step 2.

[0298] Step 2: 2-(4-hydroxy-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide (compound of formula V) A solution of 3.2 equivalents of dimethylamine (2.0 M in THF) and 3.2 equivalents of triethylamine was placed in a dropwise column. The dimethylamine solution was slowly added to the crude Step 1 reaction mixture with stirring at 5°C. The exotherm was controlled (liquid temperature <10°C). The reaction mixture was warmed to 20°C and stirred for 3 hours. IPC showed less than 5% of the remaining Step 1 intermediate. n-Heptane (13.3 V) was slowly added, cooled to 5°C, and stirred for approximately 1 hour. The slurry was filtered, and any retained solids were redissolved in 4 V of 2-MeTHF based on the wet cake mass. The 2-MeTHF / product solution was filtered to remove salts and then concentrated to 5 V. The organic layer was washed with 5 V of 5% brine to purge the tetramethyloxalamide impurity, followed by 5 V of HP water. The organic layer was then dried over sodium sulfate. n-Heptane (10 V) was slowly added to precipitate the product. The product was filtered and dried in a vacuum oven to yield intermediate (V).

[0299] HPLC: see Figure 2; 1 H-NMR (300 MHz, DMSO-D6): See Figure 3

[0300] Step 3: 3-(2-(dimethylamino)ethyl)-1H-indol-4-ol (psilocin, compound of formula II) To a THF solution of 2-(4-hydroxy-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide (V), 2 equivalents of a 1.0 M lithium aluminum hydride (LiAlH4) solution in tetrahydrofuran (THF) was added under reflux in THF for 4 hours to give 3-(2-(dimethylamino)ethyl)-1H-indol-4-ol (psilocin).

[0301] 1 H-NMR (300 MHz, DMSO-D6): See Figure 4

[0302] Step 4: Di-tert-butyl(3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate (compound of formula VI) 1 g of di-tertbutylphosphite was dissolved in 5 V of THF and added to 1.2 equivalents of N-chlorosuccinimide (NCS) / 5 V of THF at 5° C. to generate chlorodi-tertbutylphosphite in situ.

[0303] 1 H-NMR (300 MHz, CDCl); see Figure 5

[0304] Treatment of silosin with lithium diisopropylamide and a catalytic amount of DMAP in THF at 0° C., followed by dropwise addition of crude chlorodi-tert-butylphosphite, affords the corresponding di-tert-butyl(3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate.

[0305] Step 5: Psilocybin 6M hydrogen chloride (HCl) in acetone is added to crude di-tert-butyl (3-(2-(dimethylamino)ethyl)-1H-indol-4-yl) phosphate; (ii) the reaction mass is stirred and maintained at room temperature for 2 hours; (iii) the reaction mass is then neutralized with a base to a pH of 6-7; (iv) free impurities are removed from the reaction mass by extraction into an organic solvent; (v) crude psilocybin base is extracted into the organic solvent; (vi) the solvent is distilled to provide a crude psilocybin solution in the organic solvent; the crude psilocybin solution is isolated by filtration; (vii) the crude psilocybin is dissolved in an aqueous medium and neutralized with a weak acid; (viii) pure psilocybin base is extracted into the solvent; (ix) the solvent is partially distilled and psilocybin is crystallized from the same solvent; and (x) pure psilocybin is isolated by filtration.

[0306] Example 2: Exemplary Preparation of Psilocin (II) and Psilocybin (I)

[0307] [ka]

[0308] Step 1: 3-(2-chloro-2-oxoacetyl)-1H-indol-4-yl 2-chloro-2-oxoacetate (compound of formula IV) 3.0 equivalents of oxalyl chloride was added to the diethyl ether solution of unprotected 4-hydroxyindole and stirred at 0° C. for 16 hours to give 3-(2-chloro-2-oxoacetyl)-1H-indol-4-yl 2-chloro-2-oxoacetate (IV) (same as in FIG. 1, see HPLC).

[0309] Step 2: 2-(4-hydroxy-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide (compound of formula V) An excess (5.0 equiv.) of dimethylamine hydrochloride was added to a diethyl ether / pyridine solution of crude 3-(2-chloro-2-oxoacetyl)-1H-indol-4-yl 2-chloro-2-oxoacetate (2) at room temperature for 30 min to provide exclusively 2-(4-hydroxy-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide (V).

[0310] HPLC: same as Figure 2; 1 H-NMR (300 MHz, DMSO-D6): Same as in Figure 3

[0311] Step 3: 3-(2-(dimethylamino)ethyl)-1H-indol-4-ol (psilocin, compound of formula II) To a THF solution of 2-(4-hydroxy-1H-indol-3-yl)-N,N-dimethyl-2-oxoacetamide (V), 2 equivalents of a 1.0 M solution of lithium aluminum hydride (LiAlH) in tetrahydrofuran (THF) was added under reflux in THF for 4 hours to give 3-(2-(dimethylamino)ethyl)-1H-indol-4-ol (psilocin).

[0312] 1 H-NMR (300 MHz, DMSO-D6): Same as in Figure 4

[0313] Step 4: Di-tert-butyl(3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate (compound of formula VI) 1.1 equivalents of di-tert-butyl phosphite was added to sodium hydroxide (2.0 equivalents) in a 1:1 v / v solution of carbon tetrachloride (CCl4) and tetrahydrofuran (THF) at room temperature for 16 hours to give crude di-tert-butyl(3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate.

[0314] Step 5: Psilocybin 6M hydrogen chloride (HCl) in acetone is added to crude di-tert-butyl (3-(2-(dimethylamino)ethyl)-1H-indol-4-yl) phosphate; (ii) the reaction mass is stirred and maintained at room temperature for 2 hours; (iii) the reaction mass is then neutralized with a base to a pH of 6-7; (iv) free impurities are removed from the reaction mass by extraction into an organic solvent; (v) crude psilocybin base is extracted into the organic solvent; (vi) the solvent is distilled to provide a crude psilocybin solution in the organic solvent; the crude psilocybin solution is isolated by filtration; (vii) the crude psilocybin is dissolved in an aqueous medium and neutralized with a weak acid; (viii) pure psilocybin base is extracted into the solvent; (ix) the solvent is partially distilled and psilocybin is crystallized from the same solvent; and (x) pure psilocybin is isolated by filtration.

[0315] Product from step (vi): HPLC: same as Figure 6; Mass spectrum: same as Figure 7; 1 H-NMR (300 MHz, CDCl): Same as in Figure 8.

[0316] Example 3: Optimization of the preparation of di-tert-butyl (3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate in the presence of a base

[0317] [Table 1]

[0318] Reaction of silosin with di-tert-butyl phosphite in the presence of various bases. Table 1 shows that the reaction of silosin with di-tert-butyl phosphite in the presence of sodium hydroxide with DMAP or sodium hydroxide in a solvent mixture of carbon tetrachloride and tetrahydrofuran (THF) provided (3-(2-(dimethylamino)ethyl)-1H-indol-4-yl)phosphate as the major product.

[0319] Example 4: Exemplary phosphorylation of indole using chlorodi-tert-butylphosphite Di-tert-butyl phosphite was dissolved in 5 V of THF and added to 1.2 equivalents of N-chlorosuccinimide (NCS) in 5 V of THF at 5 °C to generate chlorodi-tert-butyl phosphite in situ. Unprotected 4-hydroxyindole was treated with lithium diisopropylamide and a catalytic amount of DMAP in THF at 0 °C, followed by the dropwise addition of crude chlorodi-tert-butyl phosphite to give the corresponding di-tert-butyl 1H-indol-4-yl phosphate.

[0320] Although this application has been described with reference to examples, it should be understood that the claims should not be limited to the embodiments set forth in the examples, but should be accorded the broadest interpretation consistent with the description as a whole.

[0321] All publications, patents, and patent applications are incorporated by reference herein to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety. In the event that a term in this application is found to be defined differently in a document incorporated by reference herein, the definition provided herein controls for that term. Exemplary embodiments of the present invention are described below. <1> 1. A process for preparing psilocin (compound of formula II), comprising: [ka] The process is: Unprotected 4-hydroxyindole (compound of formula III)

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Claims

1. 1. A process for preparing psilocybin (compound of formula I), comprising: 【Chemical 1】 The process comprises: Psilocin (compound of formula II) 【Chemistry 2】 in the presence of a base, (a) di-tert-butyl phosphite 【Chemistry 3】 by reacting with, or Silosin (b) chlorodi-tert-butylphosphite 【Chemistry 4】 to provide a compound of formula (VI); 【Chemistry 5】 and hydrolyzing the compound of formula (VI) to provide psilocybin (the compound of formula I).

2. 2. The process of claim 1, wherein the silosin is reacted with 1.1 to 1.5 molar equivalents of the di-tert-butyl phosphite in the presence of an excess of base in an inert solvent at a temperature and for a time to react the silosin with the di-tert-butyl phosphite to provide the compound of formula (VI).

3. 3. The process of claim 2, wherein the base is selected from sodium hydroxide and sodium hydroxide in combination with dimethylaminopyridine (DMAP).

4. The process of claim 3, wherein the silosin is reacted with di-tertbutyl phosphite at 18°C to 50°C for 2 to 8 hours.

5. The process of claim 2, wherein the inert solvent is a mixture of tetrahydrofuran and carbon tetrachloride.

6. reacting silosin with chlorodi-tert-butylphosphite to provide a compound of formula (VI); The chlorodi-tert-butylphosphite is prepared in situ by reacting di-tert-butylphosphite with N-chlorosuccinimide (NCS); the process comprising reacting silosin with di-tert-butyl phosphite and N-chlorosuccinimide (NCS) to provide a compound of formula (VI), 10. The process of claim 1.

7. The process of claim 1, further comprising preparing the psilocin (compound of formula II): 【Chemistry 6】 The process comprises: Unprotected 4-hydroxyindole (compound of formula III) 【Chemistry 7】 with oxalyl chloride to provide a compound of formula (IV); 【Chemistry 8】 reacting said compound of formula (IV) with dimethylamine (HN(CH 3 ) 2 ) to provide a compound of formula (V); 【Chemistry 9】 and reducing the compound of formula (V) with a reducing reagent to provide psilocin (compound of formula II); 【Chemistry 10】 Reacting the silosin includes: Silosin is reacted with the di-tert-butyl phosphite in the presence of the base. 【Chemistry 11】 to provide the compound of formula (VI), process.

8. The process of claim 7, wherein the silosin is reacted with di-tert-butyl phosphite in a mixture of tetrahydrofuran and carbon tetrachloride in the presence of sodium hydroxide or a combination of 4-dimethylaminopyridine (DMAP) and sodium hydroxide to provide a compound of formula (VI).

9. The process of claim 1, further comprising preparing psilocin (a compound of formula II): 【Chemistry 12】 The process comprises: Unprotected 4-hydroxyindole (compound of formula III) 【Chemistry 13】 with an excess of oxalyl chloride to provide a compound of formula (IV); 【Chemistry 14】 reacting said compound of formula (IV) with an excess of dimethylamine to provide a compound of formula (V); 【Chemistry 15】 and reducing the compound of formula (V) with a reducing reagent to provide psilocin (compound of formula II); 【Chemistry 16】 Reacting the silosin includes: The silosin was reacted with chlorodi-tert-butylphosphite 【Chemistry 17】 to provide a compound of formula (VI), process.

10. The method of claim 1, wherein the chlorodi-tert-butylphosphite is prepared in situ by reacting di-tert-butylphosphite with N-chlorosuccinimide (NCS); the process comprising reacting silosin with di-tert-butyl phosphite and N-chlorosuccinimide (NCS) to provide a compound of formula (VI), 10. The process of claim 9.

11. The process of any one of claims 7 to 10, wherein the unprotected 4-hydroxyindole (compound of formula III) is reacted with oxalyl chloride in an inert solvent at a temperature and for a time for the reaction of the unprotected 4-hydroxyindole with oxalyl chloride to provide the compound of formula (IV).

12. The process of claim 11, wherein the unprotected 4-hydroxyindole (compound of formula III) is reacted with 2 to 3 molar equivalents of the oxalyl chloride, and the temperature and time for reacting the unprotected 4-hydroxyindole (compound of formula III) with the oxalyl chloride to provide the compound of formula (IV) is 0°C to 15°C and 6 hours to 24 hours.

13. The process of claim 11, wherein the unprotected 4-hydroxyindole (compound of formula III) is reacted with 2 to 3 molar equivalents of the oxalyl chloride in MBTE, and the temperature and time for reacting the unprotected 4-hydroxyindole (compound of formula III) with the oxalyl chloride to provide the compound of formula (IV) are 0°C to 10°C and 2 hours to 3 hours to provide the compound of formula (IV).

14. A method for producing dimethylamine (HN(CH 3 ) 2 ) in situ by reacting a dimethylamine acid salt with a base; The process of any one of claims 7 to 10, wherein the compound of formula (IV) is reacted with the acid salt of dimethylamine in the presence of the base to provide the compound of formula (V).

15. The method of claim 14, wherein the compound of formula (IV) is reacted with the compound of formula (V) at a temperature and for a time to provide the compound of formula (V).

15. The process of claim 14, wherein the compound of formula (IV) is reacted with an excess of the acid salt of dimethylamine in the presence of the base in an inert solvent.

16. The process of claim 15, wherein the temperature and time for reacting the compound of formula (IV) with the acid salt of dimethylamine in the presence of the base to provide the compound of formula (V) is 18°C to 25°C and 15 minutes to 2 hours.

17. The process of claim 7, wherein the dimethylamine is dimethylamine free base.

18. The process of claim 17, wherein the compound of formula (IV) is reacted with 2 to 6 molar equivalents of the dimethylamine and 2 to 4 molar equivalents of a base to provide the compound of formula (V).

19. The process of any one of claims 7 to 10, wherein the compound of formula (V) is reacted with 1.5 to 3 molar equivalents of the reducing reagent in an inert solvent at a temperature and for a time to reduce the compound of formula (V) with the reducing reagent to provide the psilocin (compound of formula II).

20. A compound of formula (VI). 【Chemistry 18】

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