WO2021159213A1

WO2021159213A1 - Method for producing an extract of mushroom and mushroom extract made therefrom

Info

Publication number: WO2021159213A1
Application number: PCT/CA2021/050154
Authority: WO
Inventors: Bhavna SOLECKI
Original assignee: Neonmind Biosciences Inc
Current assignee: Neonmind Biosciences Inc
Priority date: 2020-02-12
Filing date: 2021-02-12
Publication date: 2021-08-19
Anticipated expiration: 2022-08-12

Abstract

A method of manufacturing a mushroom extract, the mushroom extract made by the formulation, and pharmaceutical formulations comprising same.

Description

METHOD FOR PRODUCING AN EXTRACT OF MUSHROOM AND MUSHROOM

EXTRACT MADE THEREFROM

Field of the Invention

The invention relates to a method of manufacture for an extract of fungi, specifically, a mushroom extract, and the mushroom extract made therefrom. The mushroom extract may be used in a variety of pharmaceutical formulations.

Fungus are classified as a kingdom, separate from plants or animals, and include yeasts, molds, and mushrooms. One characteristic that places fungi in a different kingdom from plants and bacteria is that they have chitin in their cell walls. Strangely, they are genetically more closely relate to animals than to plants. Fungi are useful in the decomposition of organic matter, and are commonly used as food, in the form of mushrooms and truffles, as well as a leavening or fermentation agent.

Certain fungi are known to produce bioactive compounds, for example, mycotoxins such as alkaloids and polyketides, which, though toxic at certain levels, may prove to be valuable pharmaceutic agents. Psychotropic mushrooms are also known. Mushrooms are known to produce an enormous range of natural products with antimicrobial or other biological activities, and many species have long been used, or are being developed or investigated, as sources of antibiotics, vitamins, anti-cancer compounds, and cholesterol lowering drugs. Penicillin, for example, was originally sourced from Penicillum chrysogenum , a fungi. Other antibiotics such as ciclosporin and fusidic acid are also originally sourced from fungi. Lovastatin, an HMG-CoA reductase inhibitor (or statin) can be found as a naturally occurring ingredient of the oyster mushroom. Lentinan, sourced from shitake mushroom, is approved in certain jurisdictions as a cancer treatment. Polysaccharide K, derived from fungi, is an adjuvant used in cancer therapy. Certain mushrooms have also been used for centuries in traditional medicines, such as traditional Chinese medicine.

Psilocin, or 4-hydroxy-N,N-dimethyltryptamine, and its phosphorylated prodrug, psilocybin, were first isolated from the mushroom Psilocybe Mexicana in 1958. They have both since been synthetically produced. Structural analogs of psilocin, many having higher stability, are also known, such as 4-SH-dimethyltryptamine, 1-methylpsilocin, 4-fluoro-N,N-dimethyltryptamine, O-acetylpsilocin, 4-hydroxy-N-methyl-N-isopropyltryptamine, and 4-hydroxy-N-methyl-N- ethyltryptamine.

Psilocybin

Psilocin

Psilocin/psilocybin and its analogs are most commonly used as a psychoactive recreational drug, with a half-life of 1-3 hours. Interestingly, they are known as 5-HT_2A, 5-HT_2C and 5-HT_IA agonists or partial agonists. Psilocybin is present in most psychedelic mushrooms, and at “normal” dosages creates mind-altering effects including euphoria, visual and mental allucinations, changes in perception, a distorted sense of time, spiritual experiences, nausea, and panic attacks.

Psilocin has no approved or common medical use. However, so-called “microdoses” of psilocin (typically a fraction of the hallucination-inducing dosages) have been anecedotally reported to improve attention, and there have been numerous studies performed on its potential use for treating drug dependence, anxiety, mood disorders, and depression, with varying and largely inconclusive results to date. For example, the Heffter Research Institute (heffter.org) has published research indicating that psilocybin can relieve the symptoms of anxiety and depression often found in patients with a cancer diagnosis, as well as for treatment of addiction and obsessive-compulsive disorder. Usona Institute is currently in the planning phase for a phase 2/3 clinical research program on use pf psilocybin as a treatment for major depressive disorder (www.usonaclinicaltrials.org). Psylocin and psilocybin is also used recreationally, both in small amounts (microdosing) and in larger amounts that induce hallucinatory effects. Microdose Nature (BC, Canada, www.microdosenature.com. the entirety of the site incorporated herein by reference) teaches two typical microdosing protocols: that of Dr. James Fadiman, which teaches 50-100 mg of Psilocybin cubensis fruiting bodies every third day, ideally before noon; and that of Paul Stamets of 50 - 100 mg/day for five days, followed by two days off. A microdose is typically between 10 to 400 mg of fruiting bodies per day (or an equivalent amount of purified or synthetically produced psilocin or psilocybin), with amounts higher than 400 mg per day considered “creative”, “recreational”, or hallucinatory dosages. Doses of over 5 g are generally not recommended.

Various sources of mushroom may be used; each has different potencies. One study has noted the following concentrations of psilocybin in mushrooms: Psilocybe Azurescens: 1.78%; Psilocybe Bohemica: 1.34%; Psilocybe Semilianceata: 0.98 %; Psilocybe Baeocystis 0.85%; Psilocybe Cyanescens: 0.85 %; Psilocybe Tampanensis: 0.68 %; Psilocybe Cubensis: 0.63%; Psilocybe Weilii: 0.61%, all wt/wt. There are 120 different species of mushroom within the Psilocybe genus; other genera which may contain psilocybin include Conocybe, Copelandia, Galerina, Gymnopilus, Inocybe, Mycena, Panaeolus, Pholiotina, and Pluteus.

One of the challenges in extracting active pharmaceutical ingredients from mushrooms is the cell wall. Mushroom cell walls contain glucans as well as chitin, a long chain polymer of N- acetylgluco samine .

Chitin is a relatively tough material, typically degraded by biological enzymes such as chitinase or chitinase-like proteins. Chitin creates a challenge when manufacturing mushroom extracts at an industrial scale.

Summary of the Invention

According to one aspect of the invention is provided a method for manufacturing an extract from mushrooms, comprising: obtaining a mushroom; cleaning the mushroom; drying the mushroom; chopping the mushroom; heating chopped, dry mushroom in aqueous solvent to form an aqueous fraction and a solid fraction; decanting the aqueous fraction from the solid fraction; filtering the aqueous fraction; concentrating the filtered aqueous fraction or alternatively the pre-filtered aqueous fraction; adding an alcohol solvent to the solid fraction and heating to form an alcohol fraction and a solid waste product; decanting the alcohol fraction; filtering the alcohol fraction; concentrating the filtered alcohol fraction or alternatively the pre-filtered alcohol fraction; adding the filtered alcohol fraction to the filtered aqueous fraction to obtain the mushroom oil extract.

According to certain embodiments, the chopping is to sections of about 0.01-8 cm. According to certain embodiments, the chopping is to sections of about 1.5mm.

According to certain embodiments, the aqueous solvent is purified water.

According to certain embodiments, the alcohol solvent is ethanol.

According to certain embodiments, the method further comprises wet milling and/or sonicating the chopped, dry mushroom in aqueous solvent prior to heating.

According to certain embodiments, the filtration is through a filter with a pore size of 25 microns.

According to a further aspect of the invention is provided a mushroom extract manufactured by the method hereindescribed.

According to a further aspect of the invention is provided a dry powder mushroom extract manufactured by: (a) obtaining a mushroom extract manufactured by the hereindescribed method and (b) spray drying or freeze drying said mushroom extract to obtain the dry powder mushroom extract.

According to a further aspect of the invention is provided a pharmaceutical preparation for oral administration, comprising: a mushroom extract of claim 8 or a dry powder mushroom extract as hereindescribed, and a pharmaceutically acceptable carrier or excipient.

Detailed Description

Disclosed herein is a unique mushroom extract which may have unique medicinal properties, as well as a method of manufacturing the mushroom extract from mushrooms, for example, Psilocybe mexicana, and formulations and emulsions containing said mushroom extract. The mushroom extract may be in the form of an aqueous solution, or it may be a freeze or spray dried powder. The mushroom extract produced by this method can be used advantageously in therapeutic products, for a variety of purposes, and a variety of administration methods, as detailed and exemplified further below.

The mushroom extract is made as follows.

Mushrooms, preferably fresh mushrooms, are obtained. Preferably, the entire mushroom is used, though just the mushroom cap, or just the mushroom stalk, may be used in certain embodiments.

The mushrooms are thoroughly cleaned, to remove soil, pesticide, fertilizer, or other impurities, by dry shaking, mechanical manipulation, and/or cold or room temperature water spray. In certain embodiments, for example, if the mushroom is obtained straight out of the ground, a high pressure room temperature or cold water washer can be used to wash away most of the dirt and impurities. In certain other embodiments, the mushroom may be obtained from the grower/manufacturer in a dry, washed form.

In certain embodiments, the mushrooms may be washed and sterilized in a vegetable washing machine, such as a modified Sormac GWM 2500, wherein the mushrooms are intensely washed in the machine’s wash tank in a solution of 1.5% H2O2 in water, and then mixed and transported along the length of the wash tank as a result of the turbulent water flow. The mushrooms are then over-flumed out of the wash tank, onto a vibratory de-watering screen where they are sprayed off with fresh water, then gently transported forward, draining the excess water. In certain embodiments, water and moisture are then removed, using an air knife dryer at the end of the wash and drainage cycle, which uses forced air to dry the mushrooms.

In certain embodiments, the mushrooms are then thoroughly dried, preferably at room temperature and pressure, but alternatively, if a more rapid drying is desired, a dehydrator utilizing air circulation, vacuum and/or heat can be used. A heat of up to 60 degrees Celsius may be used without affecting the desired chemical properties of the extract, with even higher heat potentially possible. Preferably, the mushroom is dried to a moisture content of less than 5%.

After drying, the mushrooms are roughly chopped, to pieces with widely varying size up to about 5 cm. Preferably, the mushrooms are is cut to about 1.5mm, which may be done in a commercial food processor such as the Hobart FP400i, which offers a continuous use hopper feeding system for high volume production.

Water is added to the dry, chopped material in a ratio of about 6 litres of water for about 500 grams of dry mushrooms.

The mushrooms are then sonicated and optionally wet milled into a slurry mixture in water. For example, an Ultrasonic UP400St, with a Sonotrode S24d7 with a 7mm tip, or a Sonotrode S24d22 with a 22mm tip, can be utilized for simultaneous wet milling and sonication (sonic cavitation). For commercial applications, the Ultrasonic can be replaced with a Digital Ultrasonic Device UlPlOOOhd (Hielscher), utilizing for example a Sonotrode BS2d22 (22 mm tip) or a Sonotrode BS2d34 (34mm tip). In one embodiment, 250 g of dessicated mushrooms is combined with 500 ml of distilled water, and the cells in the mushrooms are disrupted with the ultrasound for a period of about 5 minutes. The use of sonication and the resultant cell disruption has been found to increase the quantity of recoverable desirable active compounds recovered from the mushrooms.

The mixture is then heated to 100°C for 6-48 hours, preferably more than 24 hours, preferably without agitation.

The aqueous fraction is then decanted, filtered through a 25 micron filter, and concentrated by boiling. In certain embodiments, the concentrate may be spray or freeze dried to obtain a powder extract. In certain, preferred, embodiments, the solid fraction is then ethanol extracted by adding excess ethanol (for example, 6 litres of ethanol where 6 litres of water were used in the aqueous extraction) and heating for 2-48 hours, preferably about 6 hours, at a temperature of between 60 and 100 degrees Celcius. The ethanol fraction was then decanted, filtered through a 25 micron filter, and concentrated by boiling. The ethanol fraction was then combined with the aqueous fraction to obtain the desired mushroom extract. In certain embodiments, the combined ethanol/aqueous solution may be spray or freeze dried to obtain a powder extract.

The combined ethanol and water dual extraction provides a mushroom extract with advantageous concentrations of active ingredients, and advantageous properties.

The mushroom extract produced by this method has unique and desirable pharmaceutical properties, and can be combined with other ingredients for additional benefits, and with known pharmaceutically acceptable additives, carriers, fillers and excipients for oral, inhaled, subcutaneous, intravenous, intramuscular, vaginal, rectal, or topical administration. In particular, the mushroom extract produced by this method can be used advantageously as an orally administered microdose of psilocybin for treatment of depression, treatment of addiction, or for promotion of weight loss, especially in individuals suffering from compulsive eating disorder.

The mushroom extract or dry powder can be ingested, either alone or in an edible formulation, inhaled, for example in a vaporized form, or used as a topical cream, ointment, balm, gel, or spray. Alternatively, the mushroom extract can be injected in a suitable carrier, either intravenously, subcutaneously or intramuscularly; the mushroom extract may be administered using known skin patch technologies, rectally for example in a suppository, intravaginally, or through other high permeability areas such as the nasal cavity, the inside of the cheek, or sublingually.

Claims

Claims:

1. A method for manufacturing an extract from mushrooms, comprising: a. Obtaining a mushroom; b. Cleaning the mushroom; c. Drying the mushroom; d. Chopping the mushroom; e. Heating chopped, dry mushroom in aqueous solvent to form an aqueous fraction and a solid fraction; f. Decanting the aqueous fraction from the solid fraction; g. filtering the aqueous fraction; h. Concentrating the filtered aqueous fraction; i. Adding an alcohol solvent to the solid fraction and heating to form an alcohol fraction and a solid waste product; j. Decanting the alcohol fraction; k. filtering the alcohol fraction; l. Concentrating the filtered alcohol fraction; m. adding the filtered alcohol fraction to the filtered aqueous fraction to obtain the mushroom oil extract.

2. The method of claim 1 wherein the chopping is to sections of about 0.01-8 cm.

3. The method of claim 2 wherein the chopping is to sections of about 1.5mm.

4. The method of claim 1 wherein the aqueous solvent is purified water.

5. The method of claim 1 wherein the alcohol solvent is ethanol.

6. The method of any one of the preceding claims further comprising wet milling and/or sonicating the chopped, dry mushroom in aqueous solvent prior to heating.

7. The method of any one of the preceding claims wherein the filtration is through a filter with a pore size of 25 microns.

8. A mushroom extract manufactured by the method of any preceding claim.

9. A dry powder mushroom extract manufactured by: (a) obtaining a mushroom extract manufactured by the method of any preceding claim and (b) spray drying or freeze drying said mushroom extract to obtain the dry powder mushroom extract.

10. A pharmaceutical preparation for oral administration, comprising: a. A mushroom extract of claim 8 or a dry powder mushroom extract of claim 9, and b. A pharmaceutically acceptable carrier or excipient.