阅读说明：本技术 稳定的蛋白结合的***素组合物 (Stable protein-bound cannabinoid compositions ) 是由 什穆埃尔·科恩 威廉·Z·莱维内 希蒙·莱希特 于 2018-03-09 设计创作，主要内容包括：一种生产脂肪酸—大麻素—血浆蛋白(FCP)组合物的方法,所述方法包含：将血浆蛋白或其肽部分与包含至少一种脂肪酸的补充性脂肪酸组合物接触；以及,将血浆蛋白或其部分与包含至少一种大麻素的大麻素组合物接触,从而制备了结合组合物,所述结合组合物包含FCP复合物,在所述FCP复合物中所述至少一种脂肪酸和所述至少一种大麻素与所述血浆蛋白或其部分结合。(A method of producing a fatty acid-cannabinoid-plasma protein (FCP) composition, said method comprising: contacting a plasma protein or peptide portion thereof with a supplemental fatty acid composition comprising at least one fatty acid; and contacting the plasma protein or portion thereof with a cannabinoid composition comprising at least one cannabinoid, thereby preparing a binding composition comprising an FCP complex in which the at least one fatty acid and the at least one cannabinoid are bound to the plasma protein or portion thereof.)

1. A method of producing a fatty acid-cannabinoid-plasma protein (FCP) composition, said method comprising:

Contacting a plasma protein or peptide portion thereof with a supplemental fatty acid composition comprising at least one fatty acid; and

Contacting the plasma protein or a fraction thereof with a cannabinoid composition comprising at least one cannabinoid,

thereby preparing a binding composition comprising an FCP complex in which the at least one fatty acid and the at least one cannabinoid are bound to the plasma protein, or portion thereof.

2. The method of claim 1, wherein the at least one fatty acid comprises linoleic acid.

3. The method of claim 1, wherein the at least one fatty acid consists of linoleic acid.

4. The method of any of the preceding claims, wherein the cannabinoid composition is substantially free of fatty acids.

5. The method of claim 1, further comprising: purifying the conjugate composition to retain the plasma protein or portion thereof and to remove fatty acids or cannabinoids not conjugated to the plasma protein or portion thereof such that at least 80% of the fatty acids and at least 80% of the cannabinoids in the conjugate composition are conjugated to the plasma protein or portion thereof.

6. The method of claim 5, wherein in the conjugate composition substantially all of the fatty acid and substantially all of the cannabinoid are conjugated to the plasma protein or portion thereof.

7. The method of claim 5, wherein the binding of the fatty acid and/or the cannabinoid to the plasma protein or portion thereof is non-covalent.

8. The method of claim 7, wherein the non-covalent binding is a protein-ligand interaction.

9. The method of claim 7, wherein the non-covalent binding is characterized by non-specific lipophilic and polar interactions with the hydrophobin pocket in the plasma protein or portion thereof.

10. the method of claim 1, wherein contacting the plasma protein or portion thereof with the supplemental fatty acid comprises, mixing a composition comprising the plasma protein or portion thereof with the supplemental acid composition, and incubating the mixture.

11. The method of claim 10, wherein the incubation is performed at a temperature between 25 ℃ and 40 ℃.

12. The method of claim 10, wherein the incubation is performed in a hydroalcoholic medium.

13. The method of claim 12, wherein the alcohol contained in the hydroalcoholic medium is ethanol.

14. The method of claim 12, wherein the initial alcohol concentration in the hydroalcoholic medium is between 2% and 10%.

15. The method of claim 1, wherein contacting the plasma protein or portion thereof with the cannabinoid composition comprises mixing a composition comprising the plasma protein or portion thereof with the cannabinoid composition, and incubating the mixture.

16. The method of claim 15, wherein the incubation is performed at a temperature between 25 ℃ and 40 ℃.

17. The method of claim 15, wherein the incubation is performed in a hydroalcoholic medium.

18. The method of claim 17, wherein the alcohol contained in the hydroalcoholic medium is ethanol.

19. the method of claim 17, wherein the initial alcohol concentration in the hydroalcoholic medium is between 16% and 30%.

20. The method of claim 1, wherein the binding composition is provided in an aqueous medium.

21. The method of claim 20, wherein the aqueous medium consists of water or a salt solution.

22. The method of claim 20, further comprising drying or lyophilizing the binding composition.

23. The method of claim 1, wherein the at least one cannabinoid is one of the group consisting of tetrahydrocannabinolic acid, tetrahydrocannabinol, cannabidiolic acid, cannabidiol, cannabinol, cannabigerol, tetrahydrocannabidivarin, cannabigerolic acid, cannabichromene, HU-210, WIN55,212-2, and JWH-133, or a combination of two or more thereof.

24. The method of claim 1, wherein the plasma protein or portion thereof is selected from the group consisting of albumin, lipoprotein, glycoprotein, and alpha, beta, and gamma globulin, and mixtures of one or more thereof.

25. The method of claim 1, wherein the plasma protein or portion thereof is recombinant.

26. An FCP composition produced according to the process of claim 1.

27. A method for improving the stability of plasma protein-bound cannabinoids comprising producing a FCP composition according to the method of claim 1.

Background

Cannabinoids are compounds that are active at cannabinoid receptors and other receptors in the human body. Cannabis of plant origin, also known as phytocannabinoids, is abundant in cannabis plants. A known cannabinoid present in Cannabis at higher concentrations is tetrahydrocannabinolic acid (THCA) or its decarboxylation product Tetrahydrocannabinol (THC). Tetrahydrocannabinol has been found to have psychoactive, analgesic and antioxidant effects, as well as an increase in appetite.

Although it has proven beneficial in certain indications to ingest cannabinoids for medical use, there are disadvantages. However, the amount of active ingredient in the plant parts that are ingested may vary from plant to plant. Thus, patients treated with cannabis may lack control over the appropriate administration of active cannabinoids.

Another disadvantage of smoking cannabis for medical use is that some of the components in the cannabis smoke have a negative impact. In addition to the required cannabinoids, smoke from plant matter may also contain carcinogens, and the use of cannabis in large quantities by smoking has been associated with accelerated lung decline.

Aqueous cannabinoid compositions will enable the preparation of pharmaceutical compositions of controlled quality, precise administration for human administration and rapid absorption.

Disclosure of Invention

Embodiments of the present invention provide improved compositions comprising cannabinoids. Methods of making such compositions are also provided. In particular, the compositions described herein below are stable when in aqueous compositions. Stabilization of the composition can be determined by observing the stability of the color of the composition and/or the rate of degradation of the cannabinoid and/or the maintenance of biological activity.

As used herein, an "aqueous" composition is a composition of a liquid medium having at least 50% water. As used herein, an "aqueous" solution is a solution in which the solvent is at least 50% water.

According to one aspect of the present disclosure, there is provided a method of preparing a stable fatty acid-cannabinoid-plasma protein composition ("FCP" composition), the method comprising: contacting the plasma protein with a supplemental fatty acid composition comprising at least one fatty acid; and contacting the plasma protein with a cannabinoid composition comprising at least one cannabinoid. Optionally, the fatty acids included in the supplemental fatty acid composition provide fatty acids other than those that may be included in the cannabinoid composition or that have been complexed with plasma proteins. In embodiments of the present disclosure, at least a portion, a majority, or substantially all of the plasma proteins in the FCP composition prepared according to the above method are non-covalently associated with the fatty acid and cannabinoid, resulting in the formation of an FCP complex (FCP complex). Optionally, the non-covalent binding is a protein-ligand interaction. Optionally, the non-covalent binding is characterized by non-specific lipophilic and polar interactions, e.g., with the hydrophobin pocket in plasma proteins.

In an embodiment of the present disclosure, contacting the plasma protein with the supplemental fatty acid composition comprises mixing the composition comprising the plasma protein with the supplemental acid composition and incubating the mixture. Optionally, the incubation is performed at between 25 ℃ and 40 ℃, about 25 ℃, about 30 ℃, about 37 ℃, or about 40 ℃. Optionally, the duration of incubation is 30 minutes to 2 hours, about 30 minutes, about 1 hour, or about 2 hours. Optionally, the incubation is performed in a hydroalcoholic medium in which the initial alcohol concentration of the mixture is from 2% to 10% or about 5%. Optionally, the alcohol is ethanol.

In an embodiment of the disclosure, combining the plasma protein with the cannabinoid composition comprises mixing the composition comprising the plasma protein with the cannabinoid composition and incubating the mixture. Optionally, the incubation is performed at between 25 ℃ and 40 ℃, about 25 ℃, about 30 ℃, about 37 ℃, or about 40 ℃. Optionally, the duration of incubation is 10 to 24 hours and 16 to 18 hours. Optionally, the incubation is performed in a hydroalcoholic medium in which the initial alcohol concentration of the mixture is 16% to 30% or about 20%. Optionally, the alcohol is ethanol.

In an embodiment of the present disclosure, the method further comprises retaining the plasma protein fraction from the filtering or sieving of the FCP composition, wherein the retained plasma protein fraction comprises at least one fatty acid and at least one cannabinoid.

in embodiments of the present disclosure, at least 80%, 90%, or substantially all of the fatty acid-cannabinoid-plasma protein composition comprises tetrahydrocannabinol after filtering or sieving, or substantially all of the fatty acid comprised in the fatty acid-cannabinoid-plasma protein composition is bound to the plasma protein.

In embodiments of the present disclosure, FCP compositions are provided in the form of solutions comprising water and optionally a water-miscible non-aqueous solvent. In embodiments of the present disclosure, the FCP composition is lyophilized or dried.

In an embodiment of the present disclosure, the supplemental fatty acid composition comprises at least one fatty acid, a water-miscible non-aqueous solvent, and optionally water. Optionally, the non-aqueous solvent is ethanol. Optionally, the fatty acid composition consists of at least one fatty acid.

In an embodiment of the disclosure, the cannabinoid composition comprises a cannabis extract. The cannabis extract may be a purified cannabis extract lacking non-cannabinoid extract components such as proteins, lipids, or fatty acids. Optionally, the purified cannabis extract is enriched in cannabinoids. In an embodiment of the disclosure, a cannabinoid composition is a solution comprising at least one cannabinoid, a water-miscible, non-aqueous solvent, and optionally water.

In an embodiment of the present disclosure, the water miscible non-aqueous solvent is ethanol.

Fatty acids are carboxylic acids with hydrocarbon chains. In an embodiment of the present disclosure, the at least one fatty acid comprises one of the following or a mixture of one or two of the following: long chain fatty acids (13 or more carbons), short chain fatty acids (up to 13 carbons); a triglyceride; polyisoprenoids, or surfactants. In an embodiment of the present disclosure, the at least one fatty acid comprises linoleic acid. In another embodiment, the at least one fatty acid consists of linoleic acid.

Optionally, the at least one cannabinoid comprises one or a combination of two or more cannabinoids selected from the group consisting of: tetrahydrocannabinolic acid (THCA) or its decarboxylation product Tetrahydrocannabinol (THC), cannabidiolic acid (CBDA) or its decarboxylation product Cannabidiol (CBD), cannabidiolic acid (CBDA), Cannabinol (CBN), Cannabigerol (CBG), Tetrahydrocannabidivarin (THCV), cannabigerolic acid (CBGA), and cannabichromene (CBC). Alternatively or additionally, the group may comprise synthetic cannabinoids, such as HU-210, WIN55,212-2 and JWH-133.

as used herein, plasma proteins refer to proteins known to act as transport carriers in the bloodstream, e.g., as transport carriers for various hydrophobic compounds (including fatty acids). The plasma protein may be derived from or purified from plasma, or a recombinant protein produced in plants, bacteria, cell lines, or other microorganisms. The plasma protein is optionally selected from: albumin, lipoprotein, glycoprotein, alpha, beta and gamma globulin, and mixtures thereof. The plasma protein may comprise a whole plasma protein or a peptide comprising a portion of a plasma protein. The plasma protein fraction may comprise a cannabinoid-adsorbing fraction. The plasma protein may be a variant of a native plasma protein.

In embodiments of the present disclosure, the FCP composition comprises at least one fatty acid in an amount sufficient to substantially prevent a color change in the stabilized composition for at least 1 hour or at least 24 hours. In a more preferred embodiment, the prevention lasts at least 4 months.

In an embodiment of the present disclosure, a method comprises: (1) pretreating a plasma protein with a supplemental fatty acid composition to produce a fatty acid-plasma protein composition; and (2) mixing the fatty acid-plasma protein composition with the cannabinoid composition to produce the FCP composition.

Optionally, the method comprises: (1) pretreating a plasma protein with a cannabinoid composition to produce a cannabinoid-plasma protein composition; and (2) mixing the cannabinoid-plasma protein composition with the supplemental fatty acid composition to produce the FCP composition.

Optionally, the method comprises: (1) mixing the supplemental fatty acid composition with the cannabinoid composition to produce a supplemental fatty acid-cannabinoid composition; and (2) contacting the plasma protein with the supplemental fatty acid-cannabinoid composition to produce the FCP composition.

According to an embodiment of the disclosure, the at least one cannabinoid is not derived from a cannabis plant.

According to an embodiment of the present disclosure, at least a portion of the at least one fatty acid is not derived from a cannabis plant.

In the discussion, unless otherwise specified, adjectives such as "substantially" and "about" modifying a condition or relational characteristic of a feature of an embodiment of the invention are understood to mean that the condition or characteristic is defined to be within a tolerance range acceptable for operation of the embodiment in its intended use. Unless otherwise indicated, the term "or" in the specification and claims is to be taken as an inclusive "or" rather than an exclusive "or" and means at least one of, or any combination of, the elements to which it is attached.

This summary is provided to introduce a selection of concepts in a simplified form that are further described below in the detailed description. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.

drawings

Non-limiting examples of embodiments of the present invention are described below with reference to the figures listed after this paragraph. Identical structures, elements or parts that appear in more than one figure are labeled with the same reference number in all figures in which they appear.

Figure 1 shows a flow diagram showing the preparation of a stabilized cannabinoid composition according to an embodiment of the disclosure;

Figure 2 shows a flow diagram showing the preparation of a stabilized cannabinoid composition according to an embodiment of the disclosure;

Fig. 3A shows a chromatogram of an FCP composition (+ L composition) prepared from tetrahydrocannabinol, albumin, and linoleic acid produced in accordance with the disclosed embodiments;

Figure 3B shows a chromatogram of a cannabinoid-plasma protein (-L composition) prepared from tetrahydrocannabinol and albumin (without linoleic acid) as a control;

FIG. 4A shows a chromatogram of the + L composition shown in FIG. 3A after storage for 4 months;

FIG. 4B shows a chromatogram of the-L composition shown in FIG. 3B after storage for 4 months;

FIG. 5A shows the results of NF- κ B luciferase reporter assay to measure tetrahydrocannabinol bioactivity in + L and-L compositions after one week of storage; and

Figure 5B shows the results of the NF- κ B luciferase reporter assay to measure thc bioactivity in the + L and-L compositions after one month storage.

Detailed Description

It has long been recognized that cannabis tobacco and the herbal cannabis plant lose potency as a source of cannabinoids during storage, the loss of potency being primarily due to the loss (by degradation or otherwise) of cannabinoids (e.g., tetrahydrocannabinol).

Plasma proteins are known to act as transport carriers for a variety of hydrophobic compounds, including fatty acids, in the blood. For example, Human Serum Albumin (HSA) is the major transporter protein for fatty acid delivery to tissues, with many fatty acid binding sites. We have previously found that binding cannabinoids to plasma proteins may improve the stability of cannabinoids in aqueous compositions. Furthermore, when the source of cannabinoids is a plant material, this association also helps to separate the cannabinoids from undesirable impurities carried from the plant material into the extract used to prepare the associated cannabinoids. Generally, the binding process involves incubating the cannabinoid and plasma protein in solution under conditions to produce a cannabinoid-protein complex, then concentrating the cannabinoid-protein complex and washing the complex to remove undesirable materials not bound to the protein, thereby forming an aqueous composition enriched in the cannabinoid-protein complex.

However, such aqueous compositions have been found to change in coloration over time, indicating instability and degradation of the composition. This degradation may be associated with an undesirable decrease in efficacy of the dissolved cannabinoids, similar to the above-identified loss in efficacy of tetrahydrocannabinol, and may further be associated with plasma protein degradation.

We have surprisingly found that aqueous compositions comprising plasma proteins and cannabinoids are more stable when cannabis extracts (e.g. cannabis smoke extract or cannabis vapour extract) are used to provide cannabinoids rather than purified cannabinoids. It was initially hypothesized that fatty acids naturally present in cannabis extracts, but which are normally depleted in purified cannabinoid preparations, contribute to improved stability. We have found that the addition of a fatty acid, for example linoleic acid, during the manufacture of an FCP composition comprising a plasma protein and a cannabinoid, as described below, can stabilise the resulting composition and reduce degradation of the cannabinoid when stored in an aqueous medium. For example, the improvement in stability is demonstrated by reducing degradation of cannabinoids, reducing the appearance of cannabinoid degradation products, reducing precipitation, and reducing flocculation of aqueous FCP compositions. The improved stability is also evidenced by the controlled reduction in the degree of cannabinoid bioactivity in aqueous FCP compositions. The stable formulations are suitable, for example, for pharmaceutical formulations such as, but not limited to, solutions for parenteral or intranasal delivery.

Fig. 1 outlines the steps in a method 100 of preparing an FCP composition according to an embodiment of the present disclosure. The method 100 comprises: mixing at least one plasma protein with at least one fatty acid to produce a fatty acid-plasma protein mixture (block 110); and, mixing the fatty acid-plasma protein mixture with at least one cannabinoid (block 120).

Alternatively, the cannabinoid can be mixed with the plasma protein prior to mixing the cannabinoid-plasma protein mixture with the fatty acid.

Fig. 2 outlines the steps in a method 200 of preparing an FCP composition according to an embodiment of the present disclosure. The method 200 comprises the following steps: preparing a pre-treated plasma protein solution by adding fatty acids to the plasma protein solution (block 210); and, mixing the pre-treated plasma protein solution with a cannabinoid solution comprising a cannabinoid and a non-aqueous solvent (block 220). The FCP composition may be in the form of an aqueous composition, or may be freeze-dried or dried to a powder or lyophilized. In some embodiments, the dried FCP composition is suspended in a medium (such as a saline solution) for injection or intranasal administration, e.g., for use as an analgesic.

Cannabinoids are usually provided in the form of cannabis plant extracts. Such extracts may contain varying amounts of fatty acids depending, at least in part, on whether or how much of the extract is purified to remove non-cannabinoid compounds. However, according to embodiments of the present disclosure, supplemental fatty acids are added in addition to any fatty acids already contained in the cannabis extract for stabilizing the fatty acid-plasma protein-cannabinoid mixture. The plasma protein preparation may also contain varying amounts of fatty acids that have been complexed therewith, depending on whether or how much of the plasma protein has been purified to remove other molecules. However, according to embodiments of the present disclosure, supplemental fatty acids are added for stabilization of the cannabinoid-plasma protein complex in addition to any fatty acids that may have complexed with the plasma protein. Furthermore, the stabilizing effect of the supplemental fatty acids persists even after purification of the fatty acid-plasma protein-cannabinoid mixture to remove "loose" fatty acids not bound to plasma proteins.