COMPOSITIONS AND USES OF PSILOCYBIN AND PSILOCIN

FIELD OF THE INVENTION

[0001] This invention is directed to crystalline forms of a compound of psilocybin and psilocin and uses of the compounds in the treatment of mental health disorders and/or other central nervous system conditions and disorders.

BACKGROUND INFORMATION

[0002] Various natural plants and fungi contain psychedelic compounds which cause changes in brain function or activity (‘neuroactive’ compounds) and can also cause individuals to experience hallucinogenic or similar experiences (‘psychedelic’ compounds). Among the most common type of naturally occurring psychedelics is psilocybin, which has been found in over 180 species of mushroom globally. Psilocybin-containing mushrooms also have other neuroactive and psychedelic compounds, the most abundant of which is its primary metabolite, psilocin. In humans psilocybin is metabolized in the gastrointestinal system, primarily to psilocin, which is then absorbed and causes the psychedelic effects.

[0003] These psychedelic effects are dose-dependent and usually include changes in visual, auditory, and cognitive experiences. These psilocybin experiences may occur because both psilocybin and psilocin are similar in structure to the neurotransmitter serotonin (5- hydroxytryptamine, 5-HT), and both bind to a variety of 5-HT receptors.

[0004] In contrast to psilocybin, psilocin is chemically unstable and rapidly oxidizes when exposed to air. For this reason, there have been very few clinical or research studies on the impact of psilocin given alone or in companson to psilocybin. Pre-chmcal studies which have been carried out suggest that psilocin has a faster rate of absorption and a more rapid onset of action than psilocybin. Psilocin binds to a different range of 5-HT receptors and at different concentrations than psilocybin and as such has a different range of actions.

[0005] Because of the different properties of psilocybin and psilocin, a compound which combines both psilocybin and psilocin may be expected to have unique and beneficial properties compared to either compound alone.

[0006] It has been suggested that psychedelics such as psilocybin and psilocin may be useful in treating certain mental health disorders and central nervous system disorders. This is thought to be especially true for disorders that are resistant to currently available treatments such as post-traumatic stress disorder (PTSD) and treatment-resistant major depressive disorder, also called treatment-resistant depression (TRD).

[0007] There is a need for new treatments for wide variety of mental health disorders and central nervous system disorders. One such treatment maybe a crystalline form of psilocybin and psilocin, especially for use in disorders resistant to current treatments.

SUMMARY

[0009] The present disclosure provides several novel compositions, as well as their uses. comprising crystalline forms of psilocin and psilocybin: psilocin psilocybin

[0010] Described and identified herein as crystalline Forms A, B, and C. Crystalline Form A may be characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 10.1°. A XRPD pattern of crystalline Form A may additionally comprise a significant peak at 20 angle of about 19.16°. A XRPD pattern of cry stalline Form A may additionally comprise significant peaks at 20 angles of about 10.74°, about 25.3°, and about 24.07°. Yet further, a XRPD pattern of crystalline Form A may additionally comprise significant peaks at 20 angles of about 14.54°, about 16.5°, about 13.44°, about 23.42°, and about 8.62°. Crystalline Form B may be characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 18.54°. A XRPD pattern of crystalline Form B may additionally comprise a significant peak at 20 angle of about 8.54°. A XRPD pattern of crystalline Form B may additionally comprise significant peaks at 20 angles of about 22.78°, about 14.27°, and about 21.12°. Yet further, a XRPD pattern of crystalline Form B may additionally comprise significant peaks at 20 angles of about 14.12°, about 10.05°, about 9.94°, about 24.94°, and about 25.02°. Crystalline Form C may be characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 9.18°. A XRPD pattern of crystalline Form C may additionally comprise a significant peak at 20 angle of about 17.95°. A XRPD pattern of crystalline Form C may additionally comprise significant peaks at 20 angles of about 10.42°, about 24.22°, and about 18.38°. Yet further, a XRPD pattern of crystalline Form C may additionally comprise significant peaks at 20 angles of about 19.82°, about 17.46°, about 14.82°, about 22.38°, and about 14.06°.

[0011] In one embodiment, the present application provides pharmaceutical composition comprising a cr stalline form of psilocin and psilocybin and a pharmaceutically acceptable excipient. In one embodiment the crystalline form is crystalline Form A, crystalline Form B or cyrystaline Form C. In one embodiment, crystalline Form A is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 10. 1°; the XRPD pattern further comprises a significant peak at a 20 angle of about 19.16°; at a 29 angles of about 10.74°, about 25.3°, and about 24 07°; and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 14.54°, about 16.5°, about 13.44°, about 23.42°, and about 8.62°. In one embodiment, crystalline Fomi A has a ratio of psilocybin to psilocin of about 1: 1. In one embodiment, crystalline Form A has a chemical purity of about 95% or greater. In some embodiments, crystalline Form A contains not more than about 5 mol% of other solid forms. In some embodiments, the composition of crystalline Form A of psilocin and psilocybin comprises about 95 mol% crystalline Form A, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form A is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form A is a salt formed between psilocin and psilocybin.

[0012] In one embodiment, crystalline Form B is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 18.54°; the XRPD pattern further comprises a significant peak at a 20 angle of about 8.54°; the XRPD pattern further comprises a significant peak at a 29 angles of about 22.78°, about 14.27°, and about 21.12° and/or the XRPD pattern further comprises a significant peak at a 29 angles of about 14.12°, about 10.05°, about 9.94°, about 24.94°, and about 25.02°. In one embodiment, crystalline Form B has ratio of psilocybin to psilocin of about 1.3:1. In one embodiment, crystalline Form B has ratio of psilocybin to psilocin of about 1: 1. In one embodiment, crystalline Form B has a chemical purity of about 95% or greater. In some embodiments, crystalline Form B contains not more than about 5 mol% of other solid forms. In some embodiments, the composition of crystalline Form B of psilocin and psilocybin comprises about 95 mol% crystalline Form B, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form B is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form B is a salt formed between psilocin and psilocybin. [0013] In one embodiment, crystalline Form C is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 9. 18°; the XRPD pattern further comprises a significant peak at a 20 angle of about 17.95°; the XRPD pattern further comprises a significant peak at a 20 angles of about 10.42°, about 24.22°, and about 18.38°; and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 19.82°, about 17.46°, about 14.82°, about 22.38°, and about 14.06°. In one embodiment, crystalline Form C has ratio of psilocybin to psilocin of about 1 : 1. In one embodiment, crystalline Form C has a chemical purity of about 95% or greater. In some embodiments, crystalline Form C contains not more than about 5 mol% of other solid forms. In some embodiments, the composition of crystalline Form C of psilocin and psilocybin comprises about 95 mol% crystalline Form C, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form C is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form C is a salt formed between psilocin and psilocybin.

[0014] In one embodiment, the present invention provides a method of treating or ameliorating a disease or disorder by administering to a subject a composition comprising a crystalline form of psilocin and psilocybin thereby treating the disease or disorder

[0015] In one embodiment, the disease or disorder is a mental health disorder or a central nervous system (CNS) disorder. In one embodiment, the mental health disorder is selected from depressive disorders, anxiety disorders, post-traumatic stress disorder and addiction disorders. In one embodiment, the CNS disorder is selected from chronic pain disorders and cognitive disorders.

[0016] In one embodiment, the present invention provides a method of stimulating neurogenesis or neurite growth comprising administering to a subject a composition comprising a crystalline form of psilocin and psilocybin, thereby stimulating neurogenesis or neurite growth. In some embodiments, stimulating neurogenesis or neurite growth results in an increase in neuronal length.

DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 shows an X-ray powder diffractogram (XRPD) plot of crystalline Form A.

[0018] Figure 2 shows an XRPD plot of crystalline Form B.

[0019] Figure 3 shows an XRPD plot of crystalline Form C. [0020] Figure 4 shows the thermogravimetric mass spectroscopy (TGMS) thermogram plot of crystalline Form A.

[0021] Figure 5 shows the differential scanning calorimetry (DSC) thermogram plot of crystalline Form A.

[0022] Figure 6 shows the TGMS thermogram plot of crystalline Form B.

[0023] Figure 7 shows the DSC thermogram plot of crystalline Form B.

[0024] Figure 8 shows TGMS thermogram plot of crystalline Form C.

[0025] Figure 9 shows the DSC thermogram plot of crystalline Form C.

[0026] Figure 10 shows the 'H-NMR of crystalline Form A and psilocybin.

[0027] Figure 11 shows the H-NMR of crystalline Form A and psilocybin between 6 ppm and 7.5 ppm.

[0028] Figure 12 shows the H-NMR of crystalline Form A, crystalline Form B, and psilocybin.

[0029] Figure 13 shows the H-NMR of crystalline Form A, crystalline Form B, and psilocybin between 6 ppm and 7.5 ppm.

[0030] Figure 14 shows the dynamic vapor sorption (DVS) isotherm of crystalline Form A.

[0031] Figure 15 shows the DVS isotherm of crystalline Form B.

[0032] Figure 16 shows the DVS isotherm of crystalline Form C.

[0033] Figure 17 shows the HPLC chromatogram of cry stalline Form A.

[0034] Figure 18 shows the HPLC chromatogram of crystalline Form B.

[0035] Figure 19 shows the HPLC chromatogram of crystalline Form C.

[0036] Figure 20 shows the molecular structure and atom numbering scheme for Psilocin- Psilocybin pair obtained from simulated annealing based on the powder diffraction data of crystalline Form A.

[0037] Figure 21 shows the crystal packing and H-Bonds scheme in crystal of crystalline Form A along direction [1 00],

[0038] Figure 22 shows the graphical representation of the final cycles of Rietveld refinement for crystalline Form A.

[0039] Figure 23 shows the h5HT2a-dependent Ca2+ release. Compound activity was quantified as Ca ²⁺ release from CHO-K1 cells stably expressing h5HT2a and treated with 0.1 nM - 10 pM (left) isolated compounds; or (right) a 1 :1 equimolar combination of psilocin and psilocybin.

[0040] Figure 24 shows the h5HT2a-dependent Parrestin2 recruitment. Compound activity was quantified as |3arrestin2 recruitment from CHO-K1 cells expressing h5HT2a and treated with 0.1 nM - 10 pM compounds.

[0041] Figure 25 shows the [ ³ H]ketanserin binding, compound affinity was estimated using membranes CHO-K1 cells stably expressing h5HT2a and incubated with 0.1 nM - 10 pM compounds for 2 h.

[0042] Figure 26 shows the physiological changes in C57BL/6 mice. Compound activity was quantified in C57BL/6 mice for catalepsy (10 min) (top), nociception in the tail flick latency test (15 min) (middle), and body temperature relative to baseline (20 min) (bottom).

[0043] Figure 27 shows the head-twitch response in C57BL/6 mice, modeled rodent hallucination was quantified in C57BL/6 mice from 0-10 min after compound administration (i.p.) with the time courses of head twitches observed per minute being presented for LSD (top), psilocin (middle), and psilocybin (bottom).

[0044] Figure 28 shows the Open field test in C57BL/6 mice, compound activity was quantified in C57BL/6 mice for the OFT 30 min after treatment for total distance travelled (top, left), velocity (middle, left), and time in the center quadrant (bottom, left) with representative movement traces from Ethovision in male C57BL/6 mice (right).

[0045] Figure 29 shows the EPM in C57BL/6 mice, compound activity was quantified in C57BL/6 mice for the EPM 30 min after treatment for total distance travelled and velocity (top, left), time in each arm of the EPM (bottom, left), number of arm entries (bottom, right). Representative movement traces from Ethovision in male C57BL/6 mice (top, right).

[0046] Figure 30 shows the pharmacokinetic quantification of psilocin in C57BL/6 mouse plasma. HPLC-MS/MS method validation to date has included internal standard selection (top left), voltage optimization (top right), mobile phase organize solvent optimization (upper middle), fragmentation and standard curve creation (lower middle) with an example chromatogram from mouse plasma is shown on the bottom panel.

[0047] Figure 31 shows the EdU+/NeuN+/EdU+ data after treatment with psilocybin and/or psilocin.

[0048] Figure 32 shows the Ki67+ data after treatment with psilocybin and/or psilocin. [0049] Figure 33 shows the Sox2 positive cell data after treatment with psilocybin and/or psilocin.

[0050] Figure 34 shows the neurite outgrowth results after treatment with psilocybin and/or psilocin.

DETAILED DESCRIPTION

[0051] The present invention is based on the seminal discovery that psilocybin and psilocin form crystalline forms and specifically, that crystalline forms of a composition comprising psilocybin and psilocin may be useful for treating certain mental health disorders and central nervous system disorders as well as stimulation neurogenesis.

[0052] Before the present compositions and methods are described, it is to be understood that this invention is not limited to the particular processes, formulations, compositions, or methodologies described, as these may vary. It is also to be understood that the terminology used in the description is for the purpose of describing the particular versions or embodiments only and is not intended to limit the scope of embodiments herein which will be limited only by the appended claims. Unless defined otherw ise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of embodiments herein, the preferred methods, devices, and materials are now described. All publications mentioned herein are incorporated by reference in their entirety. Nothing herein is to be construed as an admission that embodiments herein are not entitled to antedate such disclosure by virtue of prior invention.

[0053] It must also be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. [0054] The transitional term “comprising,” which is synonymous with “including,” “containing,” or “characterized by,” is inclusive or open-ended and does not exclude additional, unrecited elements or method steps.

[0055] As used herein, the term “consists of’ or “consisting of’ means that the composition, formulation or the method includes only the elements, steps, or ingredients specifically recited in the particular claimed embodiment or claim.

[0056] As used herein, the term “consisting essentially of’ or “consists essentially of’ means that the composition, formulation or the method includes only the elements, steps or ingredients specifically recited in the particular claimed embodiment or claim and may optionally include additional elements, steps or ingredients that do not materially affect the basic and novel characteristics of the particular embodiment or claim. For example, the only active ingredient(s) in the formulation or method that treats the specified condition (e.g., nutrient depletion) is the specifically recited therapeutic(s) in the particular embodiment or claim.

[0057] As used herein, two embodiments are “mutually exclusive” when one is defined to be something which is different from the other.

[0058] When ranges of values are disclosed, and the notation “from nl ... to n2” or “between nl ... and n2” is used, where nl and n2 are the numbers, then unless otherwise specified, this notation is intended to include the numbers themselves and the range between them. This range may be integral or continuous between and including the end values. By way of example, the range “from 1 °C to 3 °C,” which is intended to include 1 °C, 3 °C, and everything in between to any number of significant figures (e.g., 1.255 °C, 2.1 °C, 2.9999 °C, etc.).

[0059] As used herein, the term “about” means plus or minus 10% of the numerical value of the number with which it is being used. Therefore, about 50% means in the range of 45% to 55%. For example, “about 100 °C” means a temperature in the range of 90°C to 110 °C. For example, “a 20 angle of about 10°” means a 29 angle in the range of 9° to 11°.

[0060] The term “substantially free” or as used herein, alone or in combination, and is used interchangeably with, the term “substantially pure”, refers to a compound which is free from all other compounds within the limits of detection as measured by any means including nuclear magnetic resonance (NMR), gas chromatography/mass spectroscopy (GC/MS), or liquid chromatography/mass spectroscopy (LC/MS). In embodiments, substantially free may be less than about 1.0%, less than about 0.5%, less than about 0.4%, less than about 0.3%, less than about 0.2%, less than about 0.1%, less than about 0.05%, or less than about 0.01%.

Crystalline Forms

[0061] The present disclosure includes embodiments directed to a composition comprising a crystalline form of psilocin and psilocybin: psilocin psilocybin

[0062] In some embodiments, the crystalline form of the composition comprising psilocin and psilocybin is crystalline Form A.

[0063] In some embodiments, crystalline Form A is characterized by a XRPD pattern comprising a significant peak at a 29 angle of about 10.1°. In some embodiments, the XRPD pattern of crystalline Form A may additionally comprise a significant peak at 20 angle of about 19.16°. In some embodiments, the XRPD pattern of crystalline Form A may additionally comprise significant peaks at 20 angles of about 10.74°, about 25.3°, and about 24.07°. In some embodiments, the XRPD pattern of crystalline Form A may additionally comprise significant peaks at 20 angles of about 14.54°, about 16.5°, about 13.44°, about 23.42°, and about 8.62°. In some embodiments, crystalline Form A is characterized by an XRPD pattern of Figure 1.

[0064] In one embodiment, crystalline Form A has a TGMS thermograph corresponding substantially to the representative TGMS thermogram as depicted in Figure 4. In some embodiments, negligible weight loss is observed. Weight loss (1.3%) is observed between 40- 200 °C by TGMS for crystalline Form A.

[0065] In one embodiment, crystalline Form A has a DSC thermogram corresponding substantially as depicted in Figure 5. In certain embodiments, crystalline Form A is characterized by a DSC plot comprising a sharp endothermic event at a temperature of about 255 °C.

[0066] In some embodiments, crystalline Form A has ratio of psilocybin to psilocin of about 1: 1, about 1.1: 1, about 1.2: 1, about 1.3: 1, about 1.4: 1, about 1.5: 1, about 1.6: 1, about 1.7: 1, about 1.8: 1, about 1.9: 1, about 2: 1, about 1: 1.1, about 1: 1.2, about 1 : 1.3, about 1 :1.4, about 1: 1.5, about 1 :1.6, about 1: 1.7, about 1: 1.8, about 1 : 1.9, or about 1 :2. In some embodiments, crystalline Form A has ratio of psilocybin to psilocin of about 1: 1.

[0067] In some embodiments, crystalline Form A has a chemical purity of about 95% or greater, is about 96% or greater, is about 97% or greater, is about 98% or greater, is about 98.5% or greater, is about 99% or greater, is about 99.5% or greater, or is about 99.8% or greater. In some embodiments, crystalline Form A is substantially pure. [0068] In some embodiments, crystalline Form A contains not more than about 0.01 mol%, about 0.02 mol%. about 0.03 mol%, about 0.04 mol%, about 0.05 mol%, about 0.06 mol%, about 0.07 mol%, about 0.08 mol%, about 0.09 mol%, about 0.1 mol%, about 0.15 mol%, about 0.2 mol%, about 0.25 mol%, about 0.3 mol%, about 0.35 mol%, about 0.4 mol%, about 0.45 mol%, about 0.5 mol%, about 0.55 mol%, about 0.6 mol%, about 0.65 mol%, about 0.7 mol%, about 0.75 mol%, about 0.8 mol%, about 0.85 mol%, about 0.9 mol%, about 0.95 mol%, about 1 mol%, about 2 mol%, about 3 mol%, about 4 mol%, about 5 mol%, about 6 mol%, about 7 mol%, about 8 mol%, about 9 mol%, about 10 mol%, about 11 mol%, about 12 mol%, about 13 mol%, about 14 mol%, about 15 mol%, about 16 mol%, about 17 mol%, about 18 mol%, about 19 mol%, about 20 mol%, of other solid forms, e.g., amorphous form. In some embodiments, crystalline Form A of is substantially free of other solid forms.

[0069] In some embodiments, the composition of crystalline Form A of psilocin and psilocybin comprises about 99 mol% crystalline Form A, about 0.5 mol% psilocin, and about 0.5 mol% psilocybin; about 98 mol% crystalline Form A, about 1.0 mol% psilocin, and about

1.0 mol% psilocybin; about 97 mol% crystalline Form A, about 1.5 mol% psilocin, and about

1.5 mol% psilocybin; about 96 mol% crystalline Form A, about 2.0 mol% psilocin, and about

2.0 mol% psilocybin; about 95 mol% crystalline Form A, about 2.5 mol% psilocin, and about

2.5 mol% psilocybin; about 94 mol% crystalline Form A, about 3.0 mol% psilocin, and about

3.0 mol% psilocybin; about 93 mol% crystalline Form A, about 3.5 mol% psilocin, and about

3.5 mol% psilocybin; about 92 mol% crystalline Form A, about 4.0 mol% psilocin, and about

4.0 mol% psilocybin; about 91 mol% crystalline Form A, about 4.5 mol% psilocin, and about

4.5 mol% psilocybin; or about 90 mol% crystalline Form A, about 5.0 mol% psilocin, and about 5.0 mol% psilocybin.

[0070] In some embodiments, crystalline Form A is anhydrous.

[0071] In some embodiments, crystalline Form A is a salt formed between psilocin and psilocybin.

[0072] In some embodiments, crystalline Form A is a co-crystal formed between psilocin and psilocybin.

[0073] Some embodiments are directed to a composition comprising a crystalline form of psilocin and psilocybin: psilocin psilocybin wherein crystalline Form A is characterized by unit cell parameters which are substantially equal to the following:

Unit cell dimensions: a = 9.3674(3) A b = 11.2660(6) A c = 24.2741(9) A a = 90 degrees

P = 90 degrees y = 90 degrees

Space group = P2i2i2i

Molecules/asymmetric unit = 1

[0074] In some embodiments, the unit cell parameters were measured about 296K.

[0075] In some embodiments, crystalline Form A has a geometry' of hydrogen bonds substantially as listed in Table A.

Table A

[0076] In some embodiments, crystalline Form A has atomic coordinates of nonhydrogen atoms substantially as listed in Table B.

Table B

[0077] In some embodiments, crystalline Form A has atomic coordinates of hydrogen atoms substantially as listed in Table C.

Table C

[0078] In some embodiments, the crystalline form of the composition comprising psilocin and psilocybin is crystalline Form B.

[0079] In some embodiments, crystalline Form B is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 18.54°. In some embodiments, crystalline Form B is characterized by a XRPD pattern of crystalline Form B may additionally comprise a significant peak at 20 angle of about 8.54°. In some embodiments, crystalline Form B is characterized by a XRPD pattern of crystalline Form B may additionally comprise significant peaks at 20 angles of about 22.78°, about 14.27°, and about 21.12°. In some embodiments, crystalline Form B is characterized by a XRPD pattern of crystalline Form B may additionally comprise significant peaks at 20 angles of about 14.12°, about 10.05°, about 9.94°, about 24.94°, and about 25.02°. In some embodiments, crystalline Form B is characterized by an XRPD pattern of Figure 2.

[0080] In one embodiment, crystalline Form B has a TGMS thermograph corresponding substantially to the representative TGMS thermogram as depicted in Figure 6. In some embodiments, negligible weight loss is observed. Weight loss (15.8%) is observed between 40- 200 °C by TGMS for crystalline Form B.

[0081] In one embodiment, crystalline Form B has a DSC thermogram corresponding substantially as depicted in Figure 7. In certain embodiments, crystalline Form B is characterized by a DSC plot comprising a broad endothermic event at a temperature of about 168.2 °C. [0082] In some embodiments, crystalline Form B has ratio of psilocybin to psilocin of about 1: 1, about 1.1: 1, about 1.2: 1, about 1.3: 1, about 1.4: 1, about 1.5: 1, about 1.6: 1, about 1.7: 1, about 1.8: 1, about 1.9: 1, about 2: 1, about 1: 1.1, about 1: 1.2, about 1 :1.3, about 1: 1.4, about 1: 1.5, about 1 :1.6, about 1: 1.7, about 1: 1.8, about 1 : 1.9, or about 1 :2. In some embodiments, crystalline Form B has ratio of psilocybin to psilocin of about 1.3: 1. In some embodiments, crystalline Form B has ratio of psilocybin to psilocin of about 1 :1.

[0083] In some embodiments, crystalline Form B has a chemical purity of about 95% or greater, is about 96% or greater, is about 97% or greater, is about 98% or greater, is about 98.5% or greater, is about 99% or greater, is about 99.5% or greater, or is about 99.8% or greater. In some embodiments, crystalline Form B is substantially pure.

[0084] In some embodiments, cry stalline Form B contains not more than about 0.01 mol%, about 0.02 mol%. about 0.03 mol%, about 0.04 mol%, about 0.05 mol%, about 0.06 mol%, about 0.07 mol%, about 0.08 mol%, about 0.09 mol%, about 0.1 mol%, about 0.15 mol%, about 0.2 mol%, about 0.25 mol%, about 0.3 mol%, about 0.35 mol%, about 0.4 mol%, about 0.45 mol%, about 0.5 mol%, about 0.55 mol%, about 0.6 mol%, about 0.65 mol%, about 0.7 mol%, about 0.75 mol%, about 0.8 mol%, about 0.85 mol%, about 0.9 mol%, about 0.95 mol%, about 1 mol%, about 2 mol%, about 3 mol%, about 4 mol%, about 5 mol%, about 6 mol%, about 7 mol%, about 8 mol%, about 9 mol%, about 10 mol%, about 11 mol%, about 12 mol%, about 13 mol%, about 14 mol%, about 15 mol%, about 16 mol%, about 17 mol%, about 18 mol%, about 19 mol%, about 20 mol%, of other solid forms, e.g., amorphous form. In some embodiments, crystalline Form B of is substantially free of other solid forms.

[0085] In some embodiments, crystalline Form B is anhydrous.

[0086] In some embodiments, crystalline Form B is a salt formed between psilocin and psilocybin.

[0087] In some embodiments, crystalline Form B is a co-crystal formed between psilocin and psilocybin.

[0088] In some embodiments, the crystalline form of the composition comprising psilocin and psilocybin is crystalline Form C.

[0089] In some embodiments, crystalline Form C is characterized by a XRPD pattern comprising a significant peak at a 29 angle of about 9.18°. In some embodiments, crystalline Form C is characterized by a XRPD pattern of crystalline Form C may additionally comprise a significant peak at 20 angle of about 17.95°. In some embodiments, crystalline Form C is characterized by a XRPD pattern of crystalline Form C may additionally comprise significant peaks at 20 angles of about 10.42°, about 24.22°, and about 18.38°. In some embodiments, crystalline Form C is characterized by a XRPD pattern of crystalline Form C may additionally comprise significant peaks at 20 angles of about 19.82°, about 17.46°, about 14.82°, about 22.38°, and about 14.06°. In some embodiments, crystalline Form C is characterized by an XRPD pattern of Figure 3.

[0090] In one embodiment, crystalline Form C has a TGMS thermograph corresponding substantially to the representative TGMS thermogram as depicted in Figure 8. In some embodiments, negligible weight loss is observed. Weight loss (13.5%) is observed between 40- 160 °C by TGMS for crystalline Form C.

[0091] In one embodiment, crystalline Form C has a DSC thennogram corresponding substantially as depicted in Figure 9. In certain embodiments, crystalline Form C is characterized by a DSC plot comprising a broad endothermic event at a temperature of about 25-140 °C, with a peak at 98.7 °C.

[0092] In some embodiments, crystalline Form C has ratio of psilocybin to psilocin of about 1: 1, about 1.1: 1, about 1.2: 1, about 1.3: 1, about 1.4: 1, about 1.5: 1, about 1.6: 1, about 1.7: 1, about 1.8: 1, about 1.9: 1, about 2: 1, about 1: 1.1, about 1: 1.2, about 1 : 1.3, about 1 :1.4, about 1: 1.5, about 1 :1.6, about 1: 1.7, about 1 : 1.8, about 1: 1.9, or about 1:2.. In some embodiments, crystalline Form C has ratio of psilocybin to psilocin of about 1 :1.

[0093] In some embodiments, crystalline Form C has a chemical purity of about 95% or greater, is about 96% or greater, is about 97% or greater, is about 98% or greater, is about 98.5% or greater, is about 99% or greater, is about 99.5% or greater, or is about 99.8% or greater. In some embodiments, crystalline Form C is substantially pure.

[0094] In some embodiments, crystalline Form C contains not more than about 0.01 mol%, about 0.02 mol%. about 0.03 mol%, about 0.04 mol%, about 0.05 mol%, about 0.06 mol%, about 0.07 mol%, about 0.08 mol%, about 0.09 mol%, about 0.1 mol%, about 0.15 mol%, about 0.2 mol%, about 0.25 mol%, about 0.3 mol%, about 0.35 mol%, about 0.4 mol%, about 0.45 mol%, about 0.5 mol%, about 0.55 mol%, about 0.6 mol%, about 0.65 mol%, about 0.7 mol%, about 0.75 mol%, about 0.8 mol%, about 0.85 mol%, about 0.9 mol%, about 0.95 mol%, about 1 mol%, about 2 mol%, about 3 mol%, about 4 mol%, about 5 mol%, about 6 mol%, about 7 mol%, about 8 mol%, about 9 mol%, about 10 mol%, about 11 mol%, about 12 mol%, about 13 mol%, about 14 mol%, about 15 mol%, about 1 mol%, about 17 mol%, about 18 mol%, about 19 mol%, about 20 mol%, of other solid forms, e.g., amorphous form. In some embodiments, crystalline Form C of is substantially free of other solid forms.

[0095] In some embodiments, crystalline Form C is anhydrous.

[0096] In some embodiments, crystalline Form C is a salt formed between psilocin and psilocybin.

[0097] In some embodiments, crystalline Form C is a co-crystal formed between psilocin and psilocybin.

Pharmaceutical Compositions

[0098] In one embodiment, the present application provides pharmaceutical composition comprising a cr stalline form of psilocin and psilocybin and a pharmaceutically acceptable excipient.

[0099] In one embodiment the crystalline form is crystalline Form A, crystalline Form B or From C. In one embodiment, crystalline Form A is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 10. 1°; the XRPD pattern further comprises a significant peak at a 20 angle of about 19.16°; at a 20 angles of about 10.74°, about 25.3°, and about 24.07°; and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 14.54°, about 16.5°, about 13.44°, about 23.42°, and about 8.62°. In one embodiment, crystalline Form A has a ratio of psilocybin to psilocin of about 1: 1. In one embodiment, crystalline Form A has a chemical purity of about 95% or greater. In some embodiments, the pharmaceutical composition of crystalline Form A contains not more than about 5 mol% of other solid forms. In some embodiments, the pharmaceutical composition of crystalline Form A of psilocin and psilocybin comprises about 95 mol% crystalline Form A, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form A is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form A is a salt formed between psilocin and psilocybin.

[0100] In one embodiment, crystalline Form A is characterized by unit cell parameters which are substantially equal to the following:

Unit cell dimensions: a = 9.3674(3) A b = 11.2660(6) A c = 24.2741(9) A a = 90 degrees P = 90 degrees y = 90 degrees Space group = P2i2i 2i Molecules/asymmetric unit = 1

[0101] In one embodiment, the unit cell parameters were measured about 296K. In one embodiment, crystalline Form A has a geometry of hydrogen bonds substantially as listed in Table A. In one embodiment, crystalline Form A has atomic coordinates of non-hydrogen atoms substantially as listed in Table B. In one embodiment, crystalline Form A has atomic coordinates of hydrogen atoms substantially as listed in Table C. In one embodiment, crystalline Form A is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form A is a salt formed between psilocin and psilocybin.

[0102] In one embodiment, crystalline Form B is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 18.54°; the XRPD pattern further comprises a significant peak at a 20 angle of about 8.54°; the XRPD pattern further comprises a significant peak at a 20 angles of about 22.78°, about 14.27°, and about 21.12° and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 14.12°, about 10.05°, about 9.94°, about 24.94°, and about 25.02°. In one embodiment, crystalline Form B has ratio of psilocybin to psilocin of about 1.3:1. In one embodiment, crystalline Form B has ratio of psilocybin to psilocin of about 1: 1. In one embodiment, crystalline Form B has a chemical purity of about 95% or greater. In some embodiments, the pharmaceutical composition of crystalline Form B contains not more than about 5 mol% of other solid forms. In some embodiments, the pharmaceutical composition of crystalline Form B of psilocin and psilocybin comprises about 95 mol% crystalline Form B, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form B is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form B is a salt formed between psilocin and psilocybin.

[0103] In one embodiment, crystalline Form C is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 9. 18°; the XRPD pattern further comprises a significant peak at a 20 angle of about 17.95°; the XRPD pattern further comprises a significant peak at a 20 angles of about 10.42°, about 24.22°, and about 18.38°; and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 19.82°, about 17.46°, about 14.82°, about 22.38°, and about 14.06°. In one embodiment, crystalline Form C has ratio of psilocybin to psilocin of about 1 : 1. In one embodiment, crystalline Form C has a chemical purity of about 95% or greater. In some embodiments, the pharmaceutical composition of crystalline Form C contains not more than about 5 mol% of other solid forms. In some embodiments, the pharmaceutical composition of crystalline Form C of psilocin and psilocybin comprises about 95 mol% crystalline Form C, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form C is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form C is a salt formed between psilocin and psilocybin.

[0104] As used herein, “pharmaceutical composition” refers to a formulation comprising an active ingredient, and optionally a pharmaceutically acceptable carrier, diluent or excipient. The term “active ingredient” can interchangeably refer to an “effective ingredient”, and is meant to refer to any agent that is capable of inducing a sought-after effect upon administration. Examples of active ingredient include, but are not limited to, chemical compound, drug, therapeutic agent, small molecule, etc. In one embodiment, the active ingredient is a crystalline form of psilocybin and psilocin, such as crystalline Form A, crystalline Form B or From C described herein.

[0105] The terms “excipient” and “pharmaceutically acceptable excipient” as used herein are intended to be generally synonymous, and is used interchangeably with, the terms “carrier,” “pharmaceutically acceptable carrier,” “diluent,” “pharmaceutically acceptable diluent.” By “pharmaceutically acceptable” it is meant the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof, nor to the activity of the active ingredient of the formulation. Pharmaceutically acceptable carriers, excipients or stabilizers are well known in the art, for example Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and may include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (for example, Zn-protein complexes); and/or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). Examples of carrier include, but are not limited to, liposome, nanoparticles, ointment, micelles, microsphere, microparticle, cream, emulsion, and gel. Examples of excipient include, but are not limited to, anti-adherents such as magnesium stearate, binders such as saccharides and their derivatives (sucrose, lactose, starches, cellulose, sugar alcohols and the like) protein like gelatin and synthetic polymers, lubricants such as talc and silica, and preservatives such as antioxidants, vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium sulfate and parabens. Examples of diluent include, but are not limited to, water, alcohol, saline solution, glycol, mineral oil and dimethyl sulfoxide (DMSO).

[0106] In one embodiment, the pharmaceutically acceptable excipient is selected from phosphate buffer; citrate buffer; ascorbic acid; methionine; octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol alcohol; butyl alcohol; benzyl alcohol; methyl paraben; propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; m-cresol; low molecular weight (less than about 10 residues) polypeptides; serum albumin; gelatin; immunoglobulins; polyvinylpyrrolidone glycine; glutamine; asparagine; histidine; arginine; lysine; monosaccharides; disaccharides; glucose; mannose; dextrins; EDTA; trehalose, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose, including eutectic solvents, eutectic-based ionic liquids, ionic liquids, sorbitol; sodium; saline; metal surfactants; non-ionic surfactants; polyethylene glycol (PEG); magnesium stearate; water; alcohol; saline solution; glycol; mineral oil and dimethyl sulfoxide (DMSO).

[0107] The active compound can be effective over a wide dosage range and can be generally administered in a therapeutically effective amount. It will be understood, however, that the amount of the compound actually administered will usually be determined by a physician, according to the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like. Method of Treatment

[0108] In one embodiment, the present invention provides a method of treating or ameliorating a disease or disorder by administering to a subject a composition comprising a crystalline form of psilocin and psilocybin thereby treating the disease or disorder.

[0109] In one embodiment, the disease or disorder is selected from a mental health disorder or a central nervous system (CNS) disorder. In one embodiment, the mental health disorder is selected from depressive disorders, anxiety disorders, post-traumatic stress disorder and addiction disorders. In one embodiment, the CNS disorder is selected from chronic pain disorders and cognitive disorders.

[0110] In one embodiment, the present invention provides a method of stimulating neurogenesis or neurite growth comprising administering to a subject a composition comprising a crystalline form of psilocin and psilocybin, thereby stimulating neurogenesis or neurite growth. In some embodiments, stimulating neurogenesis or neurite growth results in an increase in neuronal length.

[OHl] In some embodiments, stimulating neurogenesis or neurite growth results in an improvement of an impairment in cognitive or other neurological performance associated with a condition of the central nervous system. In some embodiments, the condition of the central nervous system is selected from Alzheimer's Disease, Parkinson's Disease, Huntington's Chorea, senile dementia, Pick's disease, parkinsonism dementia syndrome, progressive subcortical gliosis, progressive supranuclear palsy, thalamic degeneration syndrome, hereditary aphasia, myoclonus epilepsy, sleep deprivation, depression, stroke, ischemia, brain trauma, psychological and physical trauma, chronic pain from any cause, cognitive disorders, and cognitive deterioration.

[0112] In one embodiment the crystalline form is cr stall ine Form A, crystalline Form B, or crystalline Form C. In one embodiment, crystalline Form A is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 10.1°; the XRPD pattern further comprises a significant peak at a20 angle of about 19. 16°; at a 20 angles of about 10.74°, about 25.3°, and about 24.07°; and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 14.54°, about 16.5°, about 13.44°, about 23.42°, and about 8.62°. In one embodiment, crystalline Form A has a ratio of psilocybin to psilocin of about 1: 1. In one embodiment, cry stalline Form A has a chemical purity of about 95% or greater. In some embodiments, crystalline Form A contains not more than about 5 mol% of other solid forms. In some embodiments, the composition of crystalline Form A of psilocin and psilocybin comprises about 95 mol% crystalline Form A, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form A is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form A is a salt formed between psilocin and psilocybin.

[0113] In one embodiment, crystalline Form A is characterized by unit cell parameters which are substantially equal to the following:

Unit cell dimensions: a = 9.3674(3) A b = 11.2660(6) A c = 24.2741(9) A a = 90 degrees P = 90 degrees y = 90 degrees Space group = P2i2i2i Molecules/asymmetric unit = 1

[0114] In one embodiment, the unit cell parameters were measured about 296K. In one embodiment, crystalline Form A has a geometry of hydrogen bonds substantially as listed in Table A. In one embodiment, crystalline Form A has atomic coordinates of non-hydrogen atoms substantially as listed in Table B. In one embodiment, crystalline Form A has atomic coordinates of hydrogen atoms substantially as listed in Table C. In one embodiment, crystalline Form A is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form A is a salt formed between psilocin and psilocybin.

[0115] In one embodiment, crystalline Form B is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 18.54°; the XRPD pattern further comprises a significant peak at a 20 angle of about 8.54°; the XRPD pattern further comprises a significant peak at a 20 angles of about 22.78°, about 14.27°, and about 21.12° and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 14.12°, about 10.05°, about 9.94°, about 24.94°, and about 25.02°. In one embodiment, crystalline Form B has ratio of psilocybin to psilocin of about 1.3:1. In one embodiment, crystalline Form B has ratio of psilocybin to psilocin of about 1: 1. In one embodiment, crystalline Form B has a chemical purity of about 95% or greater. In some embodiments, crystalline Form B contains not more than about 5 mol% of other solid forms. In some embodiments, the composition of crystalline Form B of psilocin and psilocybin comprises about 92 mol% crystalline Form B, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form B is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form B is a salt formed between psilocin and psilocybin.

[0116] In one embodiment, crystalline Form C is characterized by a XRPD pattern comprising a significant peak at a 20 angle of about 9. 18°; the XRPD pattern further comprises a significant peak at a 20 angle of about 17.95°; the XRPD pattern further comprises a significant peak at a 20 angles of about 10.42°, about 24.22°, and about 18.38°; and/or the XRPD pattern further comprises a significant peak at a 20 angles of about 19.82°, about 17.46°, about 14.82°, about 22.38°, and about 14.06°. In one embodiment, crystalline Fomi C has ratio of psilocybin to psilocin of about 1 : 1. In one embodiment, crystalline Form C has a chemical purity of about 95% or greater. In some embodiments, crystalline Form C contains not more than about 5 mol% of other solid forms. In some embodiments, the composition of crystalline Form C of psilocin and psilocybin comprises about 95 mol% crystalline Form C, about 2.5 mol% psilocin, and about 2.5 mol% psilocybin. In one embodiment, crystalline Form C is a co-crystal formed between psilocin and psilocybin. In one embodiment, crystalline Form C is a salt formed between psilocin and psilocybin.

[0117] The terms "treat," "treated," "treating", or “treatment” as used herein refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder or disease, or to obtain beneficial or desired clinical results. For the purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total, whether induction of or maintenance of), whether detectable or undetectable, or enhancement or improvement of the condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment. Treatment may also be preemptive in nature, i.e., it may include prevention of disease. Prevention of a disease may involve complete protection from disease, for example as in the case of prevention of infection with a pathogen, or may involve prevention of disease progression. For example, prevention of a disease may not mean complete foreclosure of any effect related to the diseases at any level, but instead may mean prevention of the symptoms of a disease to a clinically significant or detectable level. Prevention of diseases may also mean prevention of progression of a disease to a later stage of the disease and prolonging disease- free survival as compared to disease-free survival if not receiving treatment and prolonging disease-free survival as compared to disease-free survival if not receiving treatment.

[0118] The term "treatment" is used interchangeably herein with the term "therapeutic method" and refers to both 1) therapeutic treatments or measures that cure, slow down, lessen symptoms of, and/or halt progression of a diagnosed pathologic conditions or disorder, and 2) and prophylactic/ preventative measures. Those in need of treatment may include individuals already having a particular medical disorder as well as those who may ultimately acquire the disorder (i.e., those needing preventive measures).

[0119] The terms “therapeutically effective amount”, “effective dose,” “therapeutically effective dose”, “effective amount,” or the like refer to that amount of the subject compound that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. Generally, the response is either amelioration of symptoms in a patient or a desired biological outcome (e.g., amelioration of symptoms of PTSD). The effective amount can be determined as described herein.

[0120] The terms “administration of’ and or “administering” should be understood to mean providing a pharmaceutical composition in a therapeutically effective amount to the subject in need of treatment. Administration routes can be enteral, topical or parenteral. As such, administration routes include but are not limited to intracutaneous, subcutaneous, intravenous, intraarterial, intraorbital, intracardiac, intradermal, transdermal, subarachnoid, intraspinal, oral, sublingual, buccal, rectal, nasal administrations, as well infusion. Suitable unit dosage forms, include, but are not limited to powders, tablets, pills, capsules, lozenges, suppositories, patches, nasal sprays, injectables, implantable sustained-release formulations, lipid complexes, etc.

[0121] The term “patient” is generally synonymous with the term “subject” and includes all mammals including humans. Examples of patients include humans, livestock such as cows, goats, sheep, pigs, and rabbits, and companion animals such as dogs, cats, rabbits, and horses. Preferably, the patient is a human.

[0122] Mental health disorders, or mental illness, refer to a wide range of disorders that include, but are not limited to, mood disorders, depressive disorders, mood instability disorders, mixed anxiety and depressive disorder, bipolar disorders, anxiety disorders, generalized anxiety disorders, social anxiety disorders, phobic disorders, panic attack disorders, schizophrenia, psychosis, schizoaffective disorders, eating disorders, anorexia nervosa, bulimia nervosa, substance misuse disorders, addiction disorders, alcohol abuse or dependence, opiate abuse or dependence, cocaine abuse or dependence, mixed drug abuse or dependence, behavioral addictions, gambling disorder, trauma-related disorders, post-traumatic stress disorders, grief-related disorders, loss-related disorders, end-of-life related disorders, cancer disorders, attention deficit disorder (ADD), attention deficit and hyperactivity disorder (ADHD), personality disorders, and obsessive compulsive disorders, and any combinations of these. The full list of currently accepted mental health disorders is given in the Diagnostic and Statical Manual, 5th edition (DSM5) which is incorporated by reference herein in its entirety. The central nervous system (CNS) disorders include, but are not limited to, both psychological and physical trauma following bums, injury, or any other physical cause, post-amputation, post brain-injury, post-stroke, post heart attack, post-diabetes, chronic pain disorders, migraines, chronic headaches, chronic back pain, chronic arthritic pain, chronic pain from any cause, cognitive disorders, cognitive deterioration, Alzheimer’s Disorder, dementia at any stage, dementia from any cause, age-associated memory impairment, neuropsychiatric disorders, fetal alcohol spectrum (FAS) disorders, autism spectrum disorders, multiple sclerosis, Parkinson’s disease, Fragile X, Down’s syndrome, fatigue, chronic fatigue, insomnia from any cause, and all post-covid syndromes. The severity of symptoms varies such that some individuals experience debilitating disease that precludes normal social function, while others suffer with intermittent repeated episodes across their lifespan. Although the presentation and diagnostic criteria among mental illness and CNS conditions are distinct in part, there are common endophenotypes of note across the diseases, and often comorbidities exist.

[0123] Specifically, there exist phenotypic endophenotypes associated with alterations in mood, cognition and behavior. Interestingly, many of these endophenotypes extend to neurological conditions as well. For example, attentional deficits are reported in patients with attention deficit disorder, attention deficit hyperactivity disorder, eating disorders, substance use disorders, schizophrenia, depression, obsessive compulsive disorder, traumatic brain injury, Fragile X, Alzheimer’s disease, Parkinson’s disease and frontotemporal dementia.

[0124] Depressive disorders are characterized by low mood, feeling sad or hopeless, increased irritability, sleep disturbance, lowered energy and feeling tired, poor concentration, lowered self-esteem or feeling worthless, lack of interest or pleasure in things, slowed movement, thinking they would be better off dead and self-harm, and actively considering and attempting suicide, and include major depressive disorder, bipolar depression, treatment resistant depression, and dysthymic disorder.

[0125] Anxiety disorders are characterized by feeling nervous, anxious, having difficulty breathing, avoiding people or activities, worrying excessively, physical symptoms including chest pain, palpitations of the heart, sweating, nausea, stomach ache, headache, neckache, poor sleep, poor concentration, sweating, and dizziness and include generalized anxiety disorder, social anxiety disorder, panic disorder, agoraphobia, phobias, and separation anxiety disorder. [0126] Post-traumatic stress disorder (PTSD) is characterized by flashbacks, nightmares, involuntary memories, emotional distress or physical reactions after being reminded of the trauma, avoidance of anything that can trigger such remembrances, avoiding activities, social isolation, increased anger or aggression, increased irritability, participating in risky behavior, poor concentration, problems sleeping, and an increased startle response, which can further be described as those with dissociated experiences.

[0127] Substance abuse and Addiction disorders are identified when and individual has recurrent use of alcohol or drugs which cause significant impairment in issues negatively impacting health, work, school, and home, with the severity depending upon the type and frequency of substance use, social and interpersonal problems related to this use, impact on family, friends, work, school, and relationships, withdrawal symptoms, tolerance, unsuccessful attempts to stop, physical and mental health problems from this use, and cravings, and include alcohol use disorder, cannabis use disorder, hallucinogen use disorder, inhalant use disorder, opioid use disorder, sedative hypnotic or anxiolytic use disorder, stimulant use disorder, tobacco use disorder, unknown substance use disorder, with these disorders occurring either alone or in combination.

[0128] Chronic pain disorders are characterized by pain in one or more regions of the body including headache, low back pain, neck pain, arthritis pain, pain in context of cancer, neurogenic pain, psychogenic pain, and include conditions such as chronic arthritis, osteoarthritis, chronic fatigue syndrome, endometriosis, fibromyalgia, inflammatory bowel disease, interstitial cystitis, temporomandibular joint dysfunction, vulvodynia, cluster headaches, migraine, herpes zoster, frozen shoulder, complex regional pain syndrome, gout, post-surgical pain syndrome, prolapsed intervertebral disc, sciatica, and trigeminal neuralgia. [0129] Cognitive disorders are characterized by poor functioning of the brain such that memory and other functions deteriorate including movement, speech, concentration and focus, problem-solving, analyzing sleep, tremors, tiredness, dizziness, headaches, hallucinations and confabulation, and include age-associated memory' impairment, Alzheimer’s disease, Picks disease, vascular dementia, post-stroke dementia, multi-infarct dementia, Parkinson’s disease, frontal-lobe dementia, alcohol-induced dementia, and amnesia disorders including Korsakoff’s syndrome.

[0130] Neurogenesis is a vital process in the brains of animals and humans, whereby new nerve cells are continuously generated throughout the life span of the organism. The newly bom cells are able to differentiate into functional cells of the central nervous system and integrate into existing neural circuits in the brain. Stimulating neurogenesis or neurite growth may improve cognitive performance associated with various conditions of the central nervous system such as depression and anxiety.

[0131] In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, cry stalline Form B, crystalline Form C or combinations thereof, for use in treating post-traumatic stress disorder (PTSD). In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof, for use in treating treatment resistant PTSD. In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A for use in treating treatment resistant PTSD. In one embodiment, there is provided a crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatment resistant PTSD. In one embodiment, there is provided a crystalline form of psilocybin and psilocin crystalline Form C for use in treating treatment-resistant PTSD. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form A for use in treating treatment-resistant PTSD. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatmentresistant PTSD. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin cry stalline Form C for use in treating treatment-resistant PTSD. [0132] In one embodiment, there is provided a method of treating PTSD comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, cry stalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatment-resistant PTSD comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatmentresistant PTSD comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin crystalline Form A. In one embodiment, there is provided a method of treating treatment-resistant PTSD comprising administering to a subject in need thereof an effective dose of a cry stalline form of psilocybin and psilocin crystalline Form B. In one embodiment, there is provided a method of treating treatment-resistant PTSD comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin cry stalline Form C.

[0133] In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, cry stalline Form B, crystalline Form C or combinations thereof, for use in treating depression. In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof, for use in treating treatment-resistant depression. In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A for use in treating treatmentresistant depression. In one embodiment, there is provided a crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatment-resistant depression. In one embodiment, there is provided a crystalline form of psilocybin and psilocin crystalline Form C for use in treating treatment- resistant depression. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form A for use in treating treatment-resistant depression. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatment-resistant depression. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form C for use in treating treatment-resistant depression.

[0134] In one embodiment, there is provided a method of treating depression compnsmg administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, cry stalline Form B, cry stalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatment-resistant depression comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatment-resistant depression comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin crystalline Form A. In one embodiment, there is provided a method of treating treatment-resistant depression comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin crystalline Form B. In one embodiment, there is provided a method of treating treatment-resistant depression comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin crystalline Form C.

[0135] In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof, for use in treating anxiety. In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof, for use in treating treatment-resistant anxiety. In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A for use in treating treatmentresistant anxiety. In one embodiment, there is provided a crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatment-resistant anxiety. In one embodiment, there is provided a cry stalline form of psilocybin and psilocin crystalline Form C for use in treating treatment-resistant anxiety. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form A for use in treating treatmentresistant anxiety. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatment-resistant anxiety. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form C for use in treating treatment-resistant anxiety.

[0136] In one embodiment, there is provided a method of treating anxiety comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, cry stalline Form B, cry stalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatment-resistant anxiety comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatmentresistant anxiety comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin crystalline Form A. In one embodiment, there is provided a method of treating treatment-resistant anxiety comprising admimstenng to a subject in need thereof an effective dose of a cry stalline form of psilocybin and psilocin crystalline Form B. In one embodiment, there is provided a method of treating treatment-resistant anxiety comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin cry stalline Form C.

[0137] In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof, for use in treating pain. In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof, for use in treating treatment-resistant pain. In one embodiment, there is provided a crystalline form of psilocybin and psilocin, crystalline Form A for use in treating treatment-resistant pain. In one embodiment, there is provided a crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatment-resistant pain. In one embodiment, there is provided a crystalline form of psilocybin and psilocin crystalline Form C for use in treating treatmentresistant pain. In one embodiment, there is provided a high purity cry stalline form of psilocybin and psilocin crystalline Form A for use in treating treatment-resistant pain. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form B for use in treating treatment-resistant pain. In one embodiment, there is provided a high purity crystalline form of psilocybin and psilocin crystalline Form C for use in treating treatmentresistant pain.

[0138] In one embodiment, there is provided a method of treating pain comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, cry stalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatment-resistant pain comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin, crystalline Form A, crystalline Form B, crystalline Form C or combinations thereof. In one embodiment, there is provided a method of treating treatmentresistant pain comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin crystalline Form A. In one embodiment, there is provided a method of treating treatment-resistant pain comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin crystalline Form B. In one embodiment, there is provided a method of treating treatment-resistant pain comprising administering to a subject in need thereof an effective dose of a crystalline form of psilocybin and psilocin cry stalline Form C.

[0139] In some embodiments, the composition is administered by intracutaneous, subcutaneous, intravenous, intraarterial, intradermal, transdermal, oral, sublingual, buccal, or nasal route of administration.

[0140] In some embodiments, the method further comprises the administration of a second pharmaceutically active compound.

[0141] In some embodiments, the second pharmaceutically active compound is one or more of an anti-anxiety drug, an anti-depressive drug, or an anti-pain drug.

[0142] In some embodiments, the second pharmaceutically active compound is a psychoactive alkaloid. In some embodiments the psychoactive compound or alkaloid is psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin, aeruginascin, bufotenin, bufotenidine, 5-MeO-DMT (5-methoxy-N.Ndimethyltryptamine), N,N-dimethyltryptamine (DMT), 4-hydroxytryptamine, N,N,N-trimethyl-4-hydroxytryptamine ergine (LSA), ergonovine, ergometrine, muscimol, ibotenic acid, lysergic acid hydroxyethylamide (LSH), elymoclavine, ergometrinine, or chanoclavine, or any combination thereof.

[0143] In some embodiments, the second pharmaceutically active compound is administered simultaneously with the composition.

[0144] In some embodiments, the second pharmaceutically active compound is administered sequentially with the composition.

[0145] In some embodiments, the method further comprises the administration of guided therapy.

[0146] In some embodiments, the guided therapy is administered simultaneously with the composition.

[0147] In some embodiments, the guided therapy is administered sequentially with the composition.

[0148] In some embodiments, the guided therapy is administered simultaneously with the second pharmaceutically active compound. [0149] In some embodiments, the guided therapy is administered sequentially with the second pharmaceutically active compound.

[0150] The following examples are provided to further illustrate the embodiments of the present invention, but are not intended to limit the scope of the invention. While they are typical of those that might be used, other procedures, methodologies, or techniques known to those skilled in the art may alternatively be used.

Examples

Example 1

Salt Screen [0151] Lyophilization/recrystallization: Physical mixtures of psilocin and psilocybin in a

1: 1 ratio were be prepared in Water: Acetonitrile (60:40 v/v)+0.1% formic acid. The solution was split over 11 vials to achieve 30 mg of psilocin content in each vial. The solvent was removed by freeze-drying. The obtained amorphous material was then resuspended in 10 solvents and the samples were heated to 50°C and cooled to 5°C with a constant heating rate of 10°C/h and variable cooling rates of -20°C/h, -10°C/h and -5°C/h from the first to the third cycle. After the temperature profile, the solids were harvested ambient and vacuum dried and analyzed by XPRD. All the solids were then exposed to 2 days at 40°C/75% RH (AAC) and remeasured by XRPD. Results are shown in Table 1.

Table 1 pc = poor crystalline

[0152] Slurry Conversion: Phy sical mixtures of Psilocin and Psilocybin in a molar ratio were prepared in organic solvent mixtures to form suspensions. Samples were heated to 50°C and cooled to 5°C with a constant heating rate of 10°C/h and variable cooling rates of -20°C/h, -10°C/h and -5°C/h from the first to the third cycle. Afterwards, the samples were aged at RT for 72 hours. Upon completion of the aging time, the solids were separated from the liquid phases by centrifugation and dried under ambient conditions and under vacuum before being analyzed by HT-XRPD. All solids were exposed to 2 days at 40°C/75% RH (AAC) and remeasured by XRPD. Results are shown in Table 2.

Table 2 pc = poor crystalline

Example 2 XRPD Analysis

[0153] XRPD patterns were obtained using a Ardena T2 high-throughput XRPD set-up. The plates were mounted on a Bruker General Area Detector Diffraction System (GADDS) equipped with a VANTEC-500 gas area detector corrected for intensity and geometric variations. The calibration of the measurement accuracy (peaks position) was performed using NIST SRM1976 standard (Corundum).

[0154] Data collection was carried out at room temperature using monochromatic Cu Ka radiation in the 20 region between 1.5° and 41.5°, which is the most distinctive part of the XRPD pattern. The diffraction pattern of each well was collected in two 20 ranges (1.5°< 20 < 21.5° for the first frame, and 19.5°< 20 < 41.5° for the second) with an exposure time of 90s for each frame. No background subtraction or curve smoothing was applied to the XRPD patterns.

[0155] Cell parameters as well as crystal system were obtained using LSI-Index (Coelho, 2003; Coelho & Kem, 2005) indexing program. The space group was selected on reflections condition and density of the crystal. The cell parameters, purity as well as instrument parameters were refined using Whole Powder Pattern Decomposition method.

[0156] For Rietveld calculation the cell parameters, crystal system as well as atom positions were taken from the single crystal file (cif). During the refinement the following parameters were refined: cell constants, background, instrument geometry, zero shift, and absorption.

[0157] Nor atom positions neither thermal motion parameters were refined during whole process. The following criteria of fit were used:

Yo.m. and Yc.m. are the observed and calculated data, respectively at data point m,

M the number of data points,

P the number of parameters, win the weighting given to data point m which for counting statistics is given by win = 7/o (Yo,m)2 where <s(Yo,m) is the error in Yo.m.

[0158] The XRPD of cry stalline Form A from Exp ID TCS2 is shown in Figure 1 with the peak table shown in Table 3.

Table 3

[0159] The XRPD of crystalline Form B from Exp ID TCS24 is shown in Figure 2 with the peak table shown in Table 4.

Table 4

[0160] The XRPD of crystalline Form C from Exp ID TCS26 is shown in Figure 3 with the peak table shown in Table 5.

Table 5

Example 3 TGMS and DSC Analysis

[0161] Mass loss due to solvent or water loss from the crystals was determined by TGA/DSC. Monitoring the sample weight, during heating in a TGA/DSC 3+ STARe system (Mettler-Toledo GmbH, Switzerland), resulted in a weight vs. temperature curve and a heat flow signal. The TGA/DSC 3+ was calibrated for temperature with samples of indium and aluminum. Samples (circa 2 mg) were weighed into 100 pL aluminum crucibles and sealed. The seals were pin-holed, and the crucibles heated in the TGA from 25 to 300°C at a heating rate of 10°C min-1. Dry N2 gas was used for purging. [0162] The gases coining from the TGA samples were analyzed by a mass spectrometer

Omnistar GSD 350 (Pfeiffer Vacuum GmbH, Germany). The latter is a quadrupole mass spectrometer, which analyzes masses in the temperature range of 0-200 amu.

[0163] Thermal events were obtained from DSC thermograms, recorded with a heat flux

DSC3+ STARe system (Mettler-Toledo GmbH, Switzerland). The DSC3+ was calibrated for temperature and enthalpy with a small piece of indium (m.p. = 156.6°C; 5Hf = 28.45 J/g) and zinc (m.p. = 419.6°C; 5Hf = 107.5 J/g). Samples (circa 2 mg) were sealed in standard 40 pL aluminum pans, pin-holed and heated in the DSC from 25°C to 300°C, at a heating rate of 10°C/min. Dry N2 gas, at a flow rate of 50 mL/min was used to purge the DSC equipment during measurement.

[0164] The TGMS analysis of crystalline Form A showed a mass loss of 1.3% between 40 and 200°C, due to residual ethanol/ water. Thermal decomposition started above 200°C. The DSC trace showed a single endothermic event at T _pea k of 255.0°C (Tonset 252.5°C) which corresponds to the melting of crystalline Form A. The TGMS thermogram of crystalline Form A is shown in Figure 4 and the DSC thermogram of crystalline Form A is shown in Figure 5.

[0165] The TGMS analysis of crystalline Form B showed a mass loss of 15.8% between 40 and 200°C, due to TFE and toluene based on the MS signal. Thermal decomposition started above 200°C. The DSC trace showed a broad endothermic event at T _pea k of 168.2°C (T _onS et 156. 1°C) which corresponds to the desolvation of crystalline Form B. The TGMS thermogram of crystalline Form B is shown in Figure 6 and the DSC thermogram of crystalline Form B is shown in Figure 7.

[0166] The TGMS analysis of crystalline Form C showed a mass loss of 13.5% between 40 and 160°C, due to water based on the MS signal. Thermal decomposition started above 160°C. The DSC trace showed a broad endothermic event between 25-140°C (T _pea k 98.7°C, Tonset 84.7°C) which corresponds to the dehydration of crystalline Form C. The TGMS thermogram of crystalline Form B is shown in Figure 8 and the DSC thermogram of crystalline Form B is shown in Figure 9.

Example 4 NMR Spectroscopy

[0167] ¹ H-NMR spectroscopy in D2O was used for compound integrity characterization of Psilocybin and Psilocybin-Psilocin polymorphs (crystalline Form A and crystalline Form B). The spectra were recorded at room temperature on a 500 MHz instrument (Bruker BioSpin GmbH) using standard pulse sequences. The data was processed with ACD Labs software Spectrus Processor 2016.2.2 (Advanced Chemistry Development Inc. Canada).

[0168] Figure 10 overlays the H-NMR of crystalline Form A and psilocybin while Figure 11 shows the 'H-NMR of crystalline Form A and psilocybin between 6 ppm and 7.5 ppm. Figure 12 overlays the 'H-NMR of crystalline Form A, crystalline Form B, and psilocybin while Figure 13 overlays the 'H-NMR of crystalline Form A, crystalline Form B, and psilocybin between 6 ppm and 7.5 ppm.

Example 5 DVS Analysis

[0169] Moisture sorption isotherms were collected on a DVS Adventure system from Surface Measurement Systems (London, UK). Sample size was circa 10 mg of solid material. A full sorption and desorption isotherm was recorded by varying the relative humidity from 40 - 95 - 0 - 40% in steps of 10% at a constant temperature of 25°C. Weight equilibration per step was set at dm/dt of 0.002%/min. Afterwards the sample was measured by HT-XRPD.

[0170] Crystalline Form A: The total mass difference between 0 - 95% RH was 3.6%, and the mass uptake at 80% RH was 2.4%, indicating that the material is moderately hygroscopic according to the EP hygroscopicity classification. The solid recovered after DVS was analyzed by XRPD analysis which confirmed that PSY1 is physically stable after the sorption-desorption cycle. The DVS isotherm of crystalline Form A is depicted in Figure 14.

[0171] Crystalline Form B: The total mass difference between 0 - 95% RH was 8.0%, and the mass uptake at 80% RH was 4.3%, indicating that the material is moderately hygroscopic according to the EP hygroscopicity classification. The solid recovered after DVS was analyzed by XRPD analysis which confirmed that PSY5 is physically stable after the sorption-desorption cycle. The DVS isotherm of crystalline Form B is depicted in Figure 15.

[0172] Crystalline Form C: The total mass difference between 0 - 95% RH was 16.4%, and the mass uptake at 80% RH was 1.8%, indicating that the material is slightly hygroscopic according to the EP hygroscopicity classification. The solid recovered after DVS was analyzed by XRPD which confirmed that crystalline Form C is physically stable after sorptiondesorption cycle. The gradual water uptake between 20-95% RH was 5.2% which corresponds to 1.5 molecules of water. The DVS isotherm of crystalline Form B is depicted in Figure 16.

Example 6

UPLC Analysis

[0173] UPLC analytical method

UPLC System:

• UPLC: Agilent 1290

• Detector 1 : UV detector set at 267 nm

• Detector 2: MSD XT in Positive Scan Mode UPLC Conditions:

Auto sampler temp. : RT

Column: ACE Avanor C18-PVP (100 x 2. 1mm; 1.7pm)

Column temp: 40°C

Gradient:

Mobile phase A: 0.7 gram ammonium formate 0.1% formic acid in water

Mobile phase B: 0.1% formic acid in methanol

Flow: 0.27 mL/min

Run time: 25 minutes

Gradient: Time [min] : Eluent A: Eluent B: 0 95% 5% 4 95% 5% 16.5 30% 70% 20 30% 70% 20.1 95% 5% 25 95% 5%

Sample preparation:

Concentration: ca. LO mg/mL

Solvent: Water: Acetonitrile (60:40 v/v) +0. 1% formic acid

Injection volume: 1 pl

[0174] Crystalline Form A: Reference solutions of the psilocin were made in concentrations of about 1 mg/ml. A solution of about 2 mg/mL of crystalline Form A was made in ACN/water 40/60 %v/%v + 0.01% formic acid. The stoichiometry (psilocybin: psilocin) of crystalline Form A was determined as 1 : 1. The HPLC chromatogram is shown in Figure 17.

[0175] Crystalline Form B: Reference solutions of the psilocin were made in concentrations of about 1 mg/ml. A solution of about 2 mg/mL of crystalline Form B was made in ACN/water 40/60 %v/%v + 0.01% formic acid. Applying the equation presented below (1), the stoichiometry (psilocybimpsilocin) of crystalline Form B was determined as 1.3: 1. The HPLC chromatogram is shown in Figure 18.

[0176] Crystalline Form C: Reference solutions of the psilocin were made in concentrations of about 1 mg/ml. A solution of about 2 mg/mL of crystalline Form C was made in ACN/water 40/60 %v/%v + 0.01% formic acid. The stoichiometry (psilocybin: psilocin) of crystalline Form C was determined as 1 :1 . The HPLC chromatogram is shown in Figure 19.

Example 7

Crystal Structure

[0177] High Resolution X-Ray Powder Diffraction (HR-XRPD): The HR-XRPD data of crystalline Form A were collected on D8 Advance diffractometer using Cu Kai radiation (1.54056 A) with germanium monochromator at RT. Diffraction data were collected in the 26 range 2. 15 - 41.5°. Detector scan on solid state LynxEye detector was performed using 0.0157° per step with 5 sec/step scan speed. The samples were measured in 8 mm long glass capillary with 0.3 mm outer diameter. Before transferring into the capillary the analyzed solids (from Example 1 Exp ID TCS2) was grounded in agate mortar till all of the visible crystals disappeared and material became fine powder.

[0178] Cell parameters as well as crystal system were obtained using LSI-Index indexing program. The space group was selected on reflections condition and density of the crystal. The cell parameters, purity as well as instrument parameters were refined using Whole Powder Pattern Decomposition method.

[0179] The structure was solved using simulated annealing techniques as implemented in TOPAS6. For both components Psilocin and Psilocybin the Z Matrix model was created using cif files from Cambridge Structural DataBase, CCDC1238288 for Psilocin and CCDC2128419 for Psilocybin.

[0180] Rietveld refinement was carried out using TOPAS6 (Bruker, 2017). All bond distances and angles were fixed and except for following torsion angles (C6-O5-P4-O2; C16- C15-C13-C14; C25-C24-N23-C22; C26-C25- C24-N23) only the rotation and translation of the molecules were allowed.

[0181] The following criteria of fit were used:

• Fo,mandFc,»iare the observed and calculated data, respectively at data point m

• M the number of data points

• P the number of parameters

• w’Mthe weighting given to data point m which for counting statistics is given by

[0182] The structure was determined using simulated annealing method with the rigid body allowing both molecules free rotate and translate within a unit cell. Additionally, torsion angles of phosphate as well as both amine groups were treated as variables. After obtaining chemically reasonable solution, the model was refined using Rietveld algorithm. [0183] Crystalline Form A crystallizes in orthorhombic primitive space group based on the reflections conditions and the final results of simulated annealing P212121 space group was chosen. Table 6 presents final crystal and refinement data for chosen model.

Table 6: Crystal data and final Rietveld parameters for solid for Psilocin-Psilocybm polymorph crystalline Form A.

[0184] The solution revealed that there is one Psilocin-Psilocybin pair in the asymmetric unit with no additional solvent or API molecules. All x, y, z positions of atoms can be found. Figure 20 presents molecular structure of Psilocin-Psilocybin in crystalline Form A. As it can be seen the phosphate group of Psilocybin directly interacts with N amine atom from the Psilocin. The crystal is held by extensive network of intermolecular hydrogen bonds in which every non protonated O or N atom serves as an acceptor, while every N-H and O-H acts as a donor. Table 7 presents geometry of all hydrogen bonds found in the analyzed crystal. Table 7: Hydrogen bonds found in the crystal of Psilocin-Psilocybin in crystalline Form A

Symmetry transformations: (i) 1-x, y-0.5, 1.5-z; (ii) x-1, y, z; (iii) -x, 0.5+y, 0.5-z; (iv) 0.5-x, -y, 0.5+z; (v) 0.5-x, 1-y, 0.5+z

[0185] The hydrogen bonds network of crystalline Form A forms a layered structure with the alternate layers built from Psilocin and Psilocybin molecules. These layers are parallel to the ab plane as it is presented in Figure 21.

[0186] Atomic coordinates and equivalent isotropic displacement parameters (A ^2x 10 ³ ) for non-hydrogen atoms of From A are shown in Table 8. U(eq) is defined as one third of the trace of the orthogonalized U’J tensor.

Table 8

[0187] Hydrogen coordinates and isotropic displacement parameters (A ^2x 10 ³ ) of From A are shown in Table 9. U(eq) is defined as one third of the trace of the orthogonalized U' ¹ tensor. Table 9

[0188] Figure 22 shows the graphical representation of the final cycles of Rietveld refinement which confirms that the model fits very well with collected diffractogram for crystalline Form A.

Example 8 Ca ²⁺ release assay

[0189] AequoScreen CHO-K1 cells stably-expressing h5HT2a contain an aequorin protein that, when bound to Ca ²⁺ , is able to catalyze the breakdown of the molecule coelenterazine to produce light. Emitted light levels are proportional to endoplasmic reticulum Ca ²⁺ release. CHO-K1 cells were treated with between 0.1 nM - 10 pM of each compound for 1 min before chemiluminescence is recorded. Resulting data are then fit to a three parameter non-linear regression to determine potency (EC50) and efficacy (Emax). Results are shown in Figure 23. Data were fit to a three-parameter non-linear regression in GraphPad (v. 9). n = 3- 29 independent experiments performed in duplicate or triplicate. Data are expressed as mean ± SEM EC 50 and Emax are reported in Table 10.

Table 10

[0190] Psilocin is a potent activator of Ca ²⁺ release. Psilocybin is demonstrably less potent and efficacious. A 1 :1 equimolar mixture of psilocin and psilocybin produces a Ca ²⁺ release response intermediate between either compound alone.

Example 9

Bai restin2 recruitment assay [0191] CHO-K1 cells express h5HT2a-smBiT and |3arrestin2-lgBiT fusion proteins. When h5HT2a and |3arrestin2 come into contact with one another the smBiT and IgBiT peptides complement each other to produce a complete NanoBit luciferase enzyme. When complete, the enzyme is able to catalyze the breakdown of the molecule coelenterazine to produce light. Emitted light levels are proportional to interactions between h5HT2a and Parrestin2. CHO-K1 cells were treated with between 0.1 nM - 10 pM of each compound for 2 h, during which period chemiluminescence is recorded. Area under the curve data for cumulative chemiluminescence are then fit to a three-parameter non-linear regression to determine potency (EC50) and efficacy (Emax). Results are shown in Figure 24. Data were fit to a three-parameter non-linear regression in GraphPad (v. 9). n = 3-13 independent experiments performed in duplicate or triplicate. Data are expressed as mean ± SEM. ECso and Emax are reported in Table 11.

Table 11

[0192] Psilocin is an activator of Parrestin2 recruitment. Psilocybin is not potent or efficacious. The potency of psilocin and psilocybin are greater in the Ca ²⁺ release assay than the (3arrestin2 recruitment assay.

Example 10

Barrestin recruitment assay

[0193] CHO-K1 cells stably-expressing h5EIT2a are grown, collected, and frozen at - 80°C. Cells are thawed as needed and subjected to pressure cavitation using a Dounce homogenizer to rupture cell membranes. Cell membranes are then collected by subjecting cell lysates to high-speed centrifugation at 20,000 x g for 30 min at 4°C. Homogenization and centrifugation are repeated to enrich the sample for receptor-containing cell membranes. CHO- KI cell membranes are the co-incubated with 1 nM [ ³ H]ketanserin and between 0.1 nM - 10 pM of each compound for 2 h. Samples are then pipetted onto filter paper before being rinsed and washed by vacuum filtration three times, dried, and incubated overnight in scintillation fluid. Remaining bound radioactivity is then read on a liquid scintillation counter and normalized to maximum LSD response (i.e. 0%). Resulting data are then fit to a three- parameter non-linear regression to determine affinity (Ki). Results are shown in Figure 25. Data were fit to a three-parameter non-linear regression in GraphPad (v. 9). n = 3-6 independent experiments. Data are expressed as mean ± SEM. EC 50 and Emax are reported in Table 12. Table 12

Example 11 Physiology: Catalepsy, Body temperature, and Nociception

[0194] C57BL/6 mice were treated with increasing doses of compounds as indicated via i.p. injection. Tests were as follows: Body temperature is taken before administration of the first dose of compound for baseline. Catalepsy - the bar holding assay, 10 min after compound administration. Mice are placed such that their forepaws clasp a 0.7 cm ring clamp positioned 4.5 cm above the surface of the testing space. The length of time the ring is held is recorded (sec). The trial is ended if the mouse turns its head or body, or makes 3 consecutive escape attempts, to a maximum of 60 sec. Nociception -15 min after compound administration the mouse’s tail is placed in 52°C water. Latency to remove the tail from water is recorded to a maximum of 20 sec. Body temperature - rectal thermometer 20 min after compound administration. Results are shown in Figure 26. Data were plotted in GraphPad (v. 9). n > 6 animals per dose. Data are expressed as mean ± SEM.

[0195] Psilocin and psilocybin do not produce catalepsy. Neither drug or a combination would be expected to produce freezing behavior or spasicity. Psilocin and psilocybin produce dose-dependent hypothermia. Finally, psilocin significantly reduces pain perception peaking at 1 mg/kg.

Example 12

Rodent modelling of hallucination: Head-twitch response (HTR)

[0196] C57BL/6 mice were treated with increasing doses of compounds as indicated via i.p. injection. Mice were placed inside a clear cylinder and video recorded for 10 min immediately after compound administration. Head-twitch was recorded by an individual blinded to treatment as a lateral, rhythmic shaking of the head distinct from grooming or vertical head nod. Results are shown in Figure 27. Data were plotted in GraphPad (v. 9). n > 3 animals per dose. Data are expressed as mean ± SEM.

[0197] Psilocin and psilocybin produce a time-dependent head-twitch response as a model of rodent hallucinator or dissociation. At several tested doses the peak effect appeared to occur 3-4 min post-compound administration. In contrast, the peak effect of LSD appeared to be around 7 min post-administration.

Example 13

Locomotor Effects: Open Field Task (OFT)

[0198] C57BL/6 mice were treated with increasing doses of compounds as indicated via i.p. injection. Mice are placed in the open field 30 min after compound administration. The open field measures 1 m x 1 m and the test lasts for 5 min. Total distance travelled, velocity, and time spent in the central quadrant of the field are recorded and analyzed using EthoVision XT (Noldus, v. 17). Results are shown in Figure 28. Data were plotted in GraphPad (v. 9). n > 4 animals per dose. Data are expressed as mean ± SEM.

[0199] Psilocin and psilocybin produce a dose-dependent decrease in movement, velocity, and time in the center of the open field with significant effects occurring for movement and velocity - but not time in center - at the highest doses tested (i. e. 7 or 10 mg/kg).

Example 14

Cognitive performance and anxiety: Elevated plus maze (EPM)

[0200] C57BL/6 mice were treated 3 mg/kg of compounds via i.p. injection. Mice are placed in the EPM 30 min after compound administration. The EPM measures 1 m x 1 m and the test lasts for 5 min. Total distance travelled, velocity, arm entries, and time in each arm are recorded and analyzed using EthoVision XT (Noldus, v. 17). Results are shown in Figure 29. Data were plotted in GraphPad (v. 9). n = 3-5 animals per treatment. Data are expressed as mean ± SEM.

[0201] From preliminary evidence that requires additional animals, 3 mg/kg i.p. psilocin significantly reduces closed arm time (p=0.029, unpaired /-test). but not open arm time (p=0.12, unpaired /-test): and no significant differences in arm entries. The data show a trend toward reduced overall movement and velocity, consistent with OFT data.

Example 15 Pharmacokinetics

[0202] Psilocin D-10 is prepared as an internal standard for the quantification of psilocin in mouse plasma samples (Figure 30, top left), compound quantification is optimized on the high performance liquid chromatography tandem mass spectroscopy (HPLC-MS/MS) (AB SciEx 4000 QTRAP®) for voltage (Figure 30, top right), organic solvent (80% methanol+0.1% formic acid, 90% methanol+0.1% formic acid, 100% methanol+0.1% formic acid, 90% acetonitrile+0. 1% formic acid, or 100% methanol+0.1% formic acid) (Figure 30, upper middle left), injection volume and flow rate, and creating a standard curve (Figure 30, upper middle right). The organic solvent chosen is 100% acetonitrile+0. 1% formic acid (Figure 30, upper middle right red line) because it displays the greatest signal-to-noise and most-narrow peak; and the best-performing injection volume and flow rate are 5 pL at 0.2 mL/min. Fragmentation patters and mass-to-charge ratios (m/z) were determined for psilocin D-10 (Figure 30, lower middle left). A standard curve was constructed for psilocin detection in mouse plasma with upper and lower quality controls set at 2.5 ng/mL and 400 ng/mL, respectively (Figure 30, lower middle right).

Example 16

Neurogenesis and plasticity in healthy brain organoids

[0203] The effects of 16, 50, 160, 500 and 1600 nM psilocybin, psilocin or psilocybin + psilocin in a 1 : 1 molar ratio after one exposure (6 hours, then washout) across three experiments were measured. Control brain organoids were used to start, with results validated in patient- derived organoids.

[0204] 3-month-old human brain organoids was exposed to a pulse of EdU 6 hours after final drug treatment. Newly bom neurons were quantified (e.g., EdU+/NeuN+/EdU+) to determine the extent of neurogenesis. Ki67+/Sox2+ co-label was used to study changes in the progenitor pool. EdU+/NeuN+/EdU+ data is shown in Figure 31, Ki67+ data is shown in Figure 32, and Sox2 positive cell data is shown if Figure 33. [0205] 3-month-old organoids were exposed to psilocybin or psilocin or psilocybin + psilocin in a 1 : 1 molar ratio for 6 hours and then spliced to small pieces and plate on Matrigel™ for 2 days. TUJ1 was used to label neurite outgrowth, which was quantified for length (pm) and number of branches. Results of the neurite outgrowth are shown in Figure 34. [0206] It was found that treatment with the psilocybin + psilocin in a 1 : 1 molar ratio combination resulted in an approximate 20% increase in neurogenesis from single dosing over six hours with a matching decrease in proliferation. Neuronal length two days after administration was observed to be greatest with the psilocybin + psilocin in a 1:1 molar ratio combination, approximately doubling at the highest dose and greater than psilocybin alone. [0207] Although the invention has been described with reference to the above examples, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention. Accordingly, the invention is limited only by the following claims.