Intranasal delivery of cannabinoids

Abstract

The present invention relates to a method of treatment in which a composition comprising cannabinoid and an amphiphilic carbohydrate, such as GCPQ, is administered by intranasal administration. The treatment method is particularly suitable for treating central nervous system diseases. The invention also relates to said composition for therapeutic use when made into a pharmaceutical composition with other ingredients.

Description

Technical Field

[0001] The present invention relates to a composition and a treatment method, wherein a composition comprising an amphiphilic carbohydrate and a cannabinoid is nasally delivered to a human or animal body. Background Art

[0002] Phytocannabinoids found in the Cannabis sativa plant have been recognized as having therapeutic potential. Many of these compounds can bind to multiple biological targets of the endogenous cannabinoid system in the body. The endogenous cannabinoid system regulates cognition, pain sensation, appetite, memory, sleep, immune function, and mood. These effects are mainly mediated by two members of the G protein-coupled receptor family, cannabinoid receptor 1 and 2 (CB1 and CB2), where the CB1 receptor regulates the central and peripheral nervous systems, as described by Millar, S.A. et al. ("Towards Better Delivery of Cannabidiol (CBD)", Pharmaceuticals, 2020). In particular, as one of the main active phytocannabinoids, cannabidiol (CBD) shows potential due to its good safety and lack of psychoactive effects that can lead to drug abuse. CBD has shown promising prospects as an analgesic, anticonvulsant, muscle relaxant, anxiolytic, and antipsychotic, and it has neuroprotective, anti-inflammatory, and antioxidant activities, as well as other uses under investigation. It has been reported that CBD has beneficial therapeutic effects on patients with inflammatory, neurodegenerative, and autoimmune diseases, as well as patients with epilepsy and cancer. Millar et al. pointed out that the phase 3 clinical trial of the Epidiolex CBD product showed that the product had a clinically significant improvement in refractory epilepsy caused by Lennox-Gastaut syndrome and Dravet syndrome (two of the most difficult-to-treat epilepsy types). In 2018, Epidiolex was approved by the FDA and became the first CBD-based product on the US market for the treatment of these two rare epilepsies.

[0003] However, the therapeutic potential of CBD and its development as an effective drug in the pharmaceutical industry are limited by its inherent properties such as low bioavailability and low water solubility. Cannabinoids generally have very low solubility in water (especially CBD is only 0.7 μg / mL), and they are highly lipophilic. Therefore, CBD is not easily absorbed orally, and large doses are required to produce a pharmacological effect. Due to the high lipophilicity of CBD (LogP 6.3), it is usually provided in the form of oil or alcohol preparations. An oil suspension approved for oral and buccal mucosal administration routes, for example, orally delivered in an oil solution and Sativex as an oral mucosal spray. However, the limitation of oral CBD delivery is first-pass metabolism. One study showed that the 7-hydroxycannabidiol (7-OH-CBD) metabolite accounted for 40% of the orally delivered CBD. Therefore, first-pass metabolism is an important obstacle to improving the bioavailability of orally administered CBD (Millar et al.).

[0004] In recent years, nasal administration has been explored as a potential brain delivery route that can bypass the blood-brain barrier (BBB) (Wang et al., "I.F. Nose-to-Brain Delivery.", J Pharmacol Exp Ther, 370:593-601, 2019). Nasal administration of drugs has the potential to prevent peripheral degradation, eliminate the drawbacks associated with oral administration, and allow targeted delivery of the drug to the site of action (the brain) while reducing plasma exposure, thus eliminating peripheral side effects.

[0005] In addition to oral administration, a composition and method for delivering cannabinoids to the human body are needed. The present invention provides a cannabinoid composition for nose-to-brain delivery of cannabinoids. Summary of the Invention

[0006] According to a first aspect of the present invention, there is provided a method of treatment, wherein a composition comprising an amphiphilic carbohydrate and a cannabinoid is administered intranasally to a human or animal body.

[0007] More specifically, the present invention provides a method of treating a human central nervous system disease, comprising administering to a human a composition comprising a cannabinoid and an amphiphilic carbohydrate, wherein the composition is administered intranasally to a human or animal body.

[0008] According to a second aspect of the present invention, there is provided a pharmaceutical composition suitable for intranasal administration, which comprises an amphiphilic carbohydrate, a cannabinoid, and one or more pharmaceutically acceptable excipients.

[0009] In the present invention, when referring to a method of treatment, it also means that the present invention encompasses a composition comprising an amphiphilic carbohydrate and a cannabinoid for use in those methods or treatments. For example, the present invention thus provides a composition comprising an amphiphilic carbohydrate and a cannabinoid for use in a method of treatment, wherein the composition is administered intranasally to a human or animal body.

[0010] More particularly, the present invention provides a composition comprising a cannabinoid and an amphiphilic carbohydrate for use in a method of treating a human central nervous system disease by intranasal administration. The disease is typically epilepsy. In another embodiment, the treatment may be for pain, anxiety, and / or an autoimmune disease.

[0011] The use of the above-mentioned composition in the preparation of a medicament for treatment is also provided.

[0012] The treatment method according to the present invention can deliver a higher concentration of therapeutic cannabinoids to the brain than conventional oral administration methods, while reducing the loss of the dose to the lungs or stomach, solving some problems encountered in the prior art. The treatment method involves administering a therapeutically effective amount of cannabinoids to the brain through the nose. The preparation can effectively treat a series of central nervous system diseases, including epilepsy and pain disorders. Another advantage is that the intranasal administration method according to the present invention also has fewer disadvantages, such as peripheral degradation. The composition can be formulated in a form that is conveniently compatible with a nasal spray, facilitating simple and effective intranasal delivery directly to the brain through the olfactory nerve. The main side effects of orally administered cannabidiol include diarrhea, vomiting, and elevated liver enzymes, and these side effects can be alleviated by using a delivery method that bypasses the gastrointestinal tract. Brief Description of the Drawings

[0013] Figure 1 : Schematic process diagram for preparing the aqueous formulation of GCPQ-CBD.

[0014] Figure 2 : (A) Calibration curve (3 - 100 μg / mL) and peak overlay diagram, which shows the linear and quantitative range of CBD, and (B) representative integration diagram in the MET-CBD preparation sample, which shows the detection result at a retention time of about 4 min. Chromatographic overlay of peaks obtained from CBD solutions with increasing concentrations, as shown in the calibration curve diagram in the inset.

[0015] Figure 3 : Colloidal properties of the CBD-GCPQ preparation, where CBD and GCPQ are prepared in a ratio of 1:10 g / g -1 (A) and 1:5 g / g -1 (B), and the preparation is resuspended in water at a concentration of 1 mg / mL or 5 mg / mL (in terms of CBD equivalent), which shows the particle size distribution and particle size (Z-average), polydispersity (PDI), and zeta potential (ZP) parameters measured by a particle sizer. The data are expressed as the mean ± standard error of the mean (SEM) of 3 measurements. Each line corresponds to a different batch of the preparation.

[0016] Figure 4 : Freshly prepared CBD-GCPQ (1:5 g / g) at a concentration of about 5 mg / mL (in terms of CBD equivalent) -1) Colloidal parameters of the aqueous formulation (n = 3) before spray drying ("pre-SD") and comparison with the rehydrated powder at the same concentration (ca. 5 mg / mL), showing particle size distribution, particle size (Z-average), and polydispersity (PDI) parameters by the intensimeter. Data are presented as mean ± standard error of the mean (SEM) of 3 replicate batches (3 measurements per batch). Each line corresponds to a different batch of the formulation.

[0017] Figure 5 A: Chart showing the D 50 particle size distribution of spray-dried CBD-GCPQ microparticles stored at room temperature and 4 °C for 0, 7, 14, and 30 days.

[0018] Figure 5 B: Chart showing the percentage (%) of spray-dried CBD-GCPQ particles smaller than 10 μm stored at room temperature and 4 °C for 0, 7, 14, and 30 days.

[0019] Figure 6 A: Chart showing the mean zeta potential values after reconstitution in water of spray-dried CBD-GCPQ particles stored at room temperature and 4 °C for 0, 7, 14, and 30 days.

[0020] Figure 6 B: Chart showing the polydispersity after reconstitution in water of spray-dried CBD-GCPQ particles stored at room temperature and 4 °C for 0, 7, 14, and 30 days.

[0021] Figure 6 C: Chart showing the CBD concentration (in mg / mL) after reconstitution in water of spray-dried CBD-GCPQ particles stored at room temperature and 4 °C for 0, 7, 14, and 30 days.

[0022] Figure 7 : (A) Calibration curve (1 - 500 ng / mL) and peak overlay plot showing the linear relationship and quantification range of spiked CBD in the mobile phase, validating the MRM method in non-biological samples; and (B) representative integration plot showing the peak area (AUP) and signal-to-noise ratio (SNR) at the lowest concentration of 1 ng / mL. Overlay chromatograms of peaks obtained from CBD solutions with increasing concentrations, as shown in the calibration curve plot in the inset.

[0023] Figure 8 A: Calibration curve of the AUC ratio of CBD in the brain versus concentration (1 - 100 ng / mL).

[0024] Figure 8 B: Calibration curve of the AUC ratio of CBD in plasma versus concentration (3 - 200 ng / mL).

[0025] Figure 9 : Colloidal stability and pH measurement results of the CBD-GCPQ (1:5) formulation prepared for in vivo administration, after rehydration at a concentration of 5 mg / mL (in terms of CBD equivalent), which show the initial pH (= 4.5 - 4.8) and the particle size distribution, particle size (Z-average), and polydispersity (PDI) parameters in terms of intensity after adjusting the pH to 5.5 - 5.6 with NaOH. Each curve represents a separate batch (n = 3).

[0026] Figure 10 A: Graph of CBD concentration over time in a rat in vivo pharmacokinetic study, which shows the CBD level in the brain within 2 hours (upper left).

[0027] Figure 10 B: Graph of CBD concentration over time in a rat in vivo pharmacokinetic study, which shows the CBD level in the olfactory bulb within 2 hours (upper right).

[0028] Figure 10 C: Graph of CBD concentration over time in a rat in vivo pharmacokinetic study, which shows the CBD level in plasma within 2 hours (bottom). Detailed Description

[0029] The present invention relates to compositions and methods of treatment, wherein a composition comprising an amphiphilic carbohydrate and a cannabinoid is delivered intranasally to a human or animal body.

[0030] In the compositions and methods of the present invention, the amphiphilic carbohydrate can form particulate aggregates. These aggregates can be formed by the aggregation of individual amphiphilic molecules and have an average particle size of 10 nm to 50 μm. The average particle size can be easily determined by microscopy or by using photon correlation spectroscopy, and is conveniently determined in an aqueous dispersion before filtration. The polymeric micelle aggregates can have a minimum average particle size of at least 10 nm, more preferably at least 30 nm, and a maximum average particle size preferably of 10 μm or less. The average particle size can be 10 nm to 50 μm, or 10 nm to 20 μm, or 10 nm to 5 μm, or 10 nm to 1 μm, or 10 nm to 500 nm, or 10 nm to 100 nm, or 20 nm to 50 μm, or 20 nm to 20 μm, or 20 nm to 5 μm, or 20 nm to 1 μm, or 20 nm to 500 nm, or 20 nm to 100 nm, or 50 nm to 50 μm, or 50 nm to 20 μm, or 50 nm to 5 μm, or 50 nm to 1 μm, or 50 nm to 500 nm, or 10 nm to 100 nm.

[0031] In the present invention, the amphiphilic carbohydrate is formulated together with the cannabinoid drug. The amphiphilic carbohydrate is capable of self-assembling into nanoparticles. The pharmaceutical composition of the present invention may comprise a nano-dispersion of nanoparticles of the amphiphilic carbohydrate and the drug.

[0032] The composition used in the method according to the present invention is preferably in the form of nanoparticles, which can be further processed to form a nano-in-microparticle composition by different methods - including spray drying (explained further below) or by granulation by addition to a powder, or by lyophilization. The nano-in-microparticle composition may be in the form of a dry powder and is preferably colloidally stable when reconstituted in an aqueous medium. Preferably, the nano-in-microparticle has a spherical morphology. The nano-in-microparticle may also have a hollow morphology, or may be oval or irregular in shape. Preferably, the average particle size of the nanoparticles in the nano-in-microparticle composition is less than 1000 nm, and even more preferably the average particle size of the nanoparticles is less than 500 nm. The average particle size may be from 10 nm to 1 μm, or from 10 nm to 750 nm, or from 10 nm to 500 nm, or from 10 nm to 250 nm, or from 20 nm to 1 μm, or from 20 nm to 750 nm, or from 20 nm to 500 nm, or from 20 nm to 250 nm, or from 50 nm to 1 μm, or from 50 nm to 750 nm, or from 50 nm to 500 nm, or from 50 nm to 250 nm.

[0033] The particle size distribution of the microparticles in the nano-in-microparticle composition can be measured according to D 10 、D 50 and D 90 . D 10 represents the point below which 10% of the particles on the distribution curve are; D 50 is the median particle size (or volume) distribution, representing the point below which 50% of the particles on the distribution curve are; D 90 represents the point below which 90% of the particles on the distribution curve are. The particle size distribution based on these parameters can also be calculated by laser scattering and / or microscopic methods such as scanning electron microscopy.

[0034] According to the present invention, the median volume distribution D 50 of the microparticles is preferably from 5 to 30 μm, more preferably from 10 to 25 μm. The D 10 particle size distribution of the microparticles can be less than 15 μm. The D 10 particle size distribution is preferably greater than 10 μm to avoid pulmonary deposition.

[0035] The median volume distribution D 50Typically 5 to 30 μm, preferably 10 to 25 μm. Usually, less than 10% of the particles are below 10 μm. In a further preferred embodiment, the polydispersity of the nanoparticles constituting the nano-microparticles is less than 0.5, preferably less than 0.2, more preferably less than 0.1. The polydispersity can be from 0.01 to 0.5, or from 0.01 to 0.3, or from 0.01 to 0.1, or from 0.05 to 0.5, or from 0.05 to 0.3, or from 0.05 to 0.1, or from 0.1 to 0.3, or from 0.1 to 0.2.

[0036] The amphiphilic carbohydrate in the composition is preferably a chitosan derivative, more preferably comprising units represented by the following general formula (I):

[0037]

[0038] where a + b + c + d = 1.000, and

[0039] a is between 0.01 and 0.970,

[0040] b is between 0.01 and 0.990,

[0041] c is between 0.001 and 0.970, and

[0042] d is between 0.01 and 0.990;

[0043] and wherein

[0044] X is a hydrophobic group;

[0045] R 1 、R 2 and R 3 are each independently selected from substituted or unsubstituted alkyl;

[0046] R 4 、R 5 、R 6 and R 10 are each independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted ether groups, or substituted or unsubstituted alkenyl;

[0047] R 7 may be present or absent, and when R 7 is present, it is an unsubstituted or substituted alkyl, unsubstituted or substituted amino group, or substituted or unsubstituted amide group;

[0048] R 8 and R 9 are each independently selected from hydrogen, and substituted or unsubstituted alkyl, substituted or unsubstituted ether groups, or substituted or unsubstituted alkenyl;

[0049] or a salt thereof.

[0050] In another embodiment of the general formula I above,

[0051] a is between 0.00 and 0.970,

[0052] b is between 0.01 and 0.990,

[0053] c is between 0.000 and 0.970, and

[0054] d is between 0.01 and 0.990; where a + b + c + d = 1.000.

[0055] In the general formula above, the a, b, c, and d units can be arranged in any order and can be ordered, partially ordered, or random. There can be more than one type of a, b, c, and d units - each type of a, b, c, and d unit includes a different R group. The * in the formula is used to indicate the continuation of the polymer chain.

[0056] In a preferred embodiment, the molar ratio of the d units is greater than 0.01 and more preferably at least 0.110, more preferably at least 0.120, more preferably at least 0.150 or in some embodiments at least 0.18. Generally, the molar ratio of the d units is 0.500 or less, and more preferably 0.350 or less.

[0057] Preferably, the molar ratio of the b units is from 0.010 to 0.800, and more preferably from 0.050 to 0.600.

[0058] Preferably, the molar ratio of the c units is from 0.0200 to 0.850, and more preferably from 0.05 to 0.550.

[0059] Preferably, the molar ratio of the a units is from 0.05 to 0.85 and more preferably from 0.10 to 0.75.

[0060] The d units provide a first portion of the monomer units, which are derivatized with a hydrophobic group; the b units provide a second portion of the monomer units, and they are derivatized with a quaternary nitrogen group. The a units provide a third set of groups of the monomer units, where the amine group is derivatized in a manner different from the first or second set. There can be more than one type of "a" group (e.g., different "a" groups can exist in terms of the R 8 and R 9 groups attached to the N atom).

[0061] The c units provide a fourth set of groups of the monomer units, where the amine group is not derivatized.

[0062] In the present invention, the hydrophobic group X is preferably selected from substituted or unsubstituted groups, which are alkyl groups (such as C 4-30(alkyl), alkenyl (such as C 4-30 alkenyl), alkynyl (such as C 4-30 alkynyl), aryl (such as C 5-20 aryl), polycyclic hydrophobic groups having more than one C 4 -C 8 ring structure (such as sterols (e.g., cholesterol)), polycyclic hydrophobic groups having more than one C 4 -C 8 heteroatom ring structure, polyoxo C 1 -C 4 alkylene groups (such as polyoxybutylene polymers), or hydrophobic polymer substituents (such as poly(lactic acid) groups, poly(lactic-co-glycolic acid) groups or poly(glycolic acid) groups). The X group can be a straight-chain, branched-chain or cyclic group. Any X group can be directly linked to the d unit (i.e., at the C2 position of the monomer unit), or linked through a functional group (such as an amino group, an acyl group or an amide group), thereby forming linkages that can be represented as X'-ring, X'-NH-, X'-CO-ring, X'CONH-ring, where X' is a hydrophobic group as defined above.

[0063] Preferred examples of the X group include groups represented by the formula: CH 3 (CH 2 ) n -CO-NH- or CH 3 (CH 2 ) n -NH-, or an unsaturated acid CH 3 (CH 2 ) p -CH=CH-(CH 2 ) q -CO-NH-, where n is from 4 to 30, more preferably from 6 to 20; p and q can be the same or different and are from 4 to 16, more preferably 4 to 14. A particularly preferred class of X substituents is linked to the chitosan monomer unit through an amide group, for example, a group represented by the formula CH 3 (CH 2 ) n CO-NH-, where n is from 2 to 28. Examples of amide groups are produced by coupling a carboxylic acid with the amino group of chitosan. Preferred examples are fatty acid derivatives CH 3 (CH 2 ) n COOH, such as those based on capric acid (n = 8), lauric acid (n = 10), myristic acid (n = 12), palmitic acid (n = 14), stearic acid (n = 16) or arachidic acid (n = 18).

[0064] In the above formulae, R 1 、R 2 and R3 Preferably independently of each other selected from substituted or unsubstituted alkyl, such as C 1-10 alkyl. R 1 , R 2 and / or R 3 can be straight-chain or branched. Preferably, R 1 , R 2 and R 3 are independently of each other selected from methyl, ethyl or propyl.

[0065] In the above formula, R 8 and R 9 are preferably independently of each other selected from hydrogen and substituted or unsubstituted alkyl, such as C 1-10 alkyl. R 8 and / or R 9 can be straight-chain or branched. Preferably, R 8 and R 9 are independently of each other selected from methyl, ethyl or propyl.

[0066] In the above formula, R 4 , R 5 , R 6 and R 10 are independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted ether groups, or substituted or unsubstituted alkenyl. Preferred R 4 , R 5 , R 6 and R 10 groups are substituted by one or more hydroxyl groups or other nonionic hydrophilic substituents. Examples of the R 4 , R 5 , R 6 and R 10 groups are represented by the following general formula: -(CH 2 ) p -OH, where p is from 1 to 10, preferably from 2 to 4; or -(CH 2 ) p -CH(CH 2 -OH) 2 , where p is from 1 to 10; or -(CH 2 ) p -C(CH 2 -OH) r , where p is from 1 to 10 and r is 3; or -(CH 2 CH 2 OH) p , where p is from 1 to 300.

[0067] The R 7 group may or may not be present in the general formula. When R 7When the group is absent, the quaternary ammonium functional group is directly attached to the monomer unit of the chitosan backbone. When R 7 is present, it can be an unsubstituted or substituted alkyl group (such as C 1-10 alkyl), for example, a group represented by the formula -(CH 2 ) n -; an amino group, for example, a group represented by the formula -NH-(CH 2 ) n -; or an amide group, for example, a group represented by the formula -NH-CO-(CH 2 ) n -, where n is from 1 to 10, preferably from 1 to 4. R 7 N + R 1 R 2 R 3 Preferred examples of the substituents are provided by coupling betaine (-OOC-CH 2 -N-(CH 3 ) 3 ) with the amine substituent of the b unit to provide an amide group, for example, in the form of: -NH-CO-CH 2 -N + R 1 R 2 R 3 provided.

[0068] As mentioned above, some of the substituents described herein can be unsubstituted or substituted by one or more other substituents, which is well known to those skilled in the art. Common examples of substituents include: halogen; hydroxyl; ether (such as C 1-7 alkoxy); formyl; acyl (such as C 1-7 alkyl acyl, C 5-20 aryl acyl); acyl halide; carboxyl; ester group; acyloxy; amido; acylamido; thioamido; tetrazolyl; amino; nitro; nitroso; azide; cyano; isocyano; cyanato; isocyanato; thiocyano; isothiocyano; mercapto; thioether group (such as C 1-7 alkylthio); sulfonic acid; sulfonic acid group; sulfone group; sulfonyloxy; sulfinyloxy; sulfamino; sulfonamino; sulfinamino; sulfamyl; sulfonamido; C 1-7 alkyl (for example, including unsubstituted C 1-7 alkyl, C1-7 haloalkyl, C 1-7 hydroxyalkyl, C 1-7 carboxyalkyl, C 1-7 aminoalkyl, C 5-20 aryl-C 1-7 alkyl); C 3-20 heterocyclic group; and C 5-20 aryl (e.g., including C 5-20 carboaryl, C 5-20 heteroaryl, C 1-7 alkyl-C 5-20 aryl and C 5-20 haloaryl) groups.

[0069] As used herein, the term "ring structure" refers to a closed ring having 3 to 10 covalently linked atoms, more preferably 3 to 8 covalently linked atoms, and even more preferably 5 to 6 covalently linked atoms. The ring can be an alicyclic or aromatic ring. As used herein, the term "alicyclic" refers to a ring that is not an aromatic ring.

[0070] As used herein, the term "carbocyclic" refers to a ring in which all ring atoms are carbon atoms.

[0071] As used herein, the term "carboaromatic ring" refers to an aromatic ring in which all ring atoms are carbon atoms.

[0072] As used herein, the term "heterocyclic" refers to a ring in which at least one ring atom is a polyvalent heteroatom (e.g., nitrogen, phosphorus, silicon, oxygen, or sulfur, but more commonly nitrogen, oxygen, or sulfur). Preferably, the heterocyclic ring has 1 to 4 heteroatoms.

[0073] The above rings can be part of a "polycyclic group".

[0074] In one embodiment, the amphiphilic carbohydrate can include an additional group, i.e., acyl A:

[0075]

[0076] The group A can be present in an amount of 0.5 mole% to 30 mole%, and the contents of the remaining units can be adjusted accordingly such that the range of a is 0.05 mole% to 40 mole%, the range of b is 5 mole% to 20 mole%, the range of c is 0.05 mole% to 20 mole%, and the range of d is 5 mole% to 30 mole%.

[0077] The preferred amphiphilic carbohydrate of the present invention has the following formula:

[0078]

[0079] Wherein:

[0080] Unit a' corresponds to unit c in Formula I;

[0081] Units b' and e' together correspond to unit a in Formula I;

[0082] Unit c' corresponds to unit b in Formula I;

[0083] Unit d' corresponds to unit d in Formula I;

[0084] Unit f' corresponds to the above-mentioned unit A;

[0085] The ratio of each unit a'+b'+c'+d'+e'+f' = 1, and the preferred content of each unit is as described above;

[0086] or a salt thereof.

[0087] Preferably, the amphiphilic carbohydrate is quaternary ammonium palmitoyl glycol chitosan (GCPQ).

[0088] Preferably, the amphiphilic carbohydrate is N-palmitoyl-N-methyl-N,N-dimethyl-N,N,N-trimethyl-6-O-glycol chitosan, also known as quaternary ammonium palmitoyl glycol chitosan (GCPQ).

[0089] In this case, the palmitoylation level of GCPQ (corresponding to group d) is preferably 5-50% per monomer, for example 10-25% per monomer. In one embodiment, the d level is less than or equal to 31% per monomer, preferably 11% to 31%, more preferably 11% to 20%.

[0090] The quaternization level (b) is preferably 3-40% per monomer, preferably 10-30% per monomer. In one embodiment, the b level is less than or equal to 17% per monomer, preferably 8% to 17%.

[0091] The indicated percentage levels are mole percentages (mole%).

[0092] The molecular weight range of the amphiphilic carbohydrate (usually GCPQ) can be 1-40 kDa or 1-30 kDa, for example 5-30 kDa, 10-30 kDa or 8-20 kDa. More preferably, the molecular weight is about 10-15 kDa.

[0093] The method according to the present invention can be used to treat infectious and autoimmune diseases of the central nervous system.

[0094] The cannabinoid is preferably a non-psychoactive cannabinoid. A non-psychoactive cannabinoid refers to a compound derived from the cannabis plant that does not cause changes in perception, behavior, or psychological processes. For example, a non-limiting list of non-psychoactive cannabinoids suitable for the present invention includes: cannabidiol (CBD), cannabinol (CBG), cannabidiolic acid (CBDA), cannabidivarin (CBDV), and cannabichromene (CBC). Preferably, the non-psychoactive cannabinoid in the present invention is cannabidiol.

[0095] Cannabinoids can be used as analgesics, anticonvulsants, muscle relaxants, anxiolytics, or antipsychotics. They may also have neuroprotective, anti-inflammatory, and antioxidant activities. Therefore, the present invention can be used for any of the above uses.

[0096] The present invention is particularly suitable for delivering non-psychoactive cannabinoids to the brain. The non-psychoactive cannabinoids delivered according to the present invention can exert a therapeutic effect in the brain. Cannabinoids have been shown to be particularly effective against central nervous system diseases, especially infectious and autoimmune diseases of the central nervous system (CNS).

[0097] The compositions according to the present invention can be used to treat a variety of diseases, including epilepsy, Dravet syndrome, Bell's palsy, cerebral palsy, Alzheimer's disease, dementia, motor neuron disease, multiple sclerosis, Parkinson's disease, long COVID, neurofibromatosis, shingles, sciatica, pain, and sleep disorders (such as migraine), Lennox-Gastaut syndrome, mental diseases, neurodegenerative diseases, and brain cancer.

[0098] According to the present invention, the preferred therapeutic indications are epilepsy, pain, anxiety, and autoimmune diseases.

[0099] The treatment method of the present invention involves intranasal delivery of a composition comprising a cannabinoid and an amphiphilic carbohydrate to an animal or human body. Intranasal administration of the drug can enhance targeted delivery to the brain while eliminating common harmful side effects that may be caused by other administration routes (such as the oral route). By bypassing the blood-brain barrier (BBB), the drug is delivered to the top of the nasal cavity and transported along the olfactory nerve, thereby achieving targeted delivery to the brain.

[0100] By using an amphiphilic carbohydrate, a cannabinoid composition can be developed and appropriately formulated into nanoparticles, or preferably nano-microparticles, to be compatible with a nasal drug delivery device for intranasal administration of the compound. Preferably, the drug is cannabidiol (CBD) and the amphiphilic carbohydrate is GCPQ. The composition may be suitable for a nasal spray. By using this specific method, the dose lost to the lungs or stomach during intranasal administration is significantly reduced.

[0101] The device can work in the manner described below.

[0102] The formulations of the present invention can be dispensed from a fluid dispenser, where the fluid is in particular a gas carrying solid or liquid particles. The dispenser has: a container for the fluid; a chamber for receiving the particles, which is in fluid communication with the container in use; means for moving the fluid from the container to the chamber; means necessary for bringing the fluid into contact with the particles contained in the chamber to (in the case of particle aggregation) de-aggregate them and agitate them into a turbulent flow so as to produce a flowing fluid containing the particles; an outlet that can be arranged in fluid communication with the chamber; and release means for releasing the flowing fluid from the dispenser through the outlet. The term "turbulent flow" as used herein includes swirling or eddy currents - both of which are preferred forms of turbulent flow. The dispenser can be used and operated in any orientation, including upright, inverted or lying flat. Another advantage is that it solves the problems of particle de-aggregation and / or fluidization using relatively simple and potentially inexpensive technology. The particles are typically solid particles and can be used to dispense particulate compositions including drugs in a metered manner, especially in the case where the particles are solid particles that tend to cake during storage and / or transportation. The properties of any particles can be any one of a variety of properties compatible with the function of the device of the present invention, such as their density, particle size, specific surface area, required dose, etc. Preferably, the particles have a narrow particle size distribution and similar shapes.

[0103] The fluid in the container can be any such liquid: which is a sufficiently flowing fluid to agitate the particles into a turbulent flow, and which is stable during storage, and which is inert to the particles and the dispensing target.

[0104] The fluid can be a gas, such as air, or, if the particles cannot be inert to air for a long time, then it can be nitrogen, a conventional optionally fluorinated lower hydrocarbon propellant such as hydrofluorocarbon (HFC) or carbon dioxide; a liquid, such as water, or, if the particles cannot be inert to water for a long time, then it can be (usually pressurized) a conventional optionally fluorinated lower hydrocarbon propellant (such as butane or HFC), a hydrofluoroalkane (HFA) propellant or any compatible combination thereof.

[0105] Fluid communication between the container and the chamber generally includes at least one channel, and preferably at least a pair of channels, which extend between the container and the chamber. Any channel extending between the container and the chamber can take the form of a conduit, a duct, or a tube. The shape and size of any channel for agitating particles by fluid can be arbitrarily selected within a wide range compatible with this function. These can depend on the specific properties of the particles, such as their density, particle size, specific surface area, required dose, etc., and can have a circular cross-section and / or any other regular curved cross-section, such as generally an elliptical, semi-circular or semi-elliptical cross-section. However, usually each channel has a linear cross-section, such as a channel with a trough-shaped, triangular, square or rectangular cross-section. For an amount of 0.5 - 35 mg of the composition including the drug provided in the fully loaded dispenser, the cross-sectional area of each channel is usually 0.03 to 3.0 mm 2 , particularly 1.0 to 1.5 mm 2 . The channel can have a variety of shapes varying along its length, such as curved, but is usually straight.

[0106] When there is at least a pair of channels, all channels generally have similar or even the same size and configuration. Of course, the channels in a pair of channels can each have the opposite handedness when appropriate.

[0107] The container is a pressurized fluid container, so when the release device is triggered, the pressurized fluid is pushed from the container into the chamber under its own pressure. In this case, the release device may be the same component as the device for allowing the pressurized fluid to enter the chamber and contact the particles. The pressurized fluid container has advantages in the dispenser of the present invention: it can provide faster and / or stronger fluid discharge into the chamber. For (in the case of particle aggregation) de-aggregating the particles contained in the chamber and agitating them into a turbulent flow to produce a flowing fluid containing particles, especially in the case where the particles are quite prone to caking during storage and / or transportation, this may be ideal.

[0108] Amphiphilic carbohydrates (such as GCPQ) are biocompatible polymers, which can encapsulate the contained drug to form a protective molecular encapsulation. Drugs (such as the non-psychoactive cannabinoids preferred in the present invention) can be efficiently loaded into the resulting stable nanoparticles.

[0109] The nanoparticle composition can be synthesized by common nanoparticle synthesis methods known in the art. This includes sol-gel method, chemical vapor deposition (CVD), atomic layer deposition (ALD), molecular layer deposition (MLD), high-pressure homogenization and other microfluidization techniques, probe sonication, simply shaking the drug in an aqueous medium in the presence of an amphiphilic carbohydrate, grinding, and rehydrating after thin film preparation. Preferably, the method includes dissolving the cannabinoid in an amphiphilic carbohydrate solution, then evaporating to form a thin film, and then further rehydrating. The ratio of non-psychoactive cannabinoid to amphiphilic carbohydrate is usually 0.5:10 to 5:10 g g -1 The ratio of cannabinoid to amphiphilic carbohydrate can be 0.5:10 to 5:10 g g -1 or 0.5:10 to 4:10 g g -1 or 0.5:10 to 3:10 g g -1 or 0.5:10 to 2:10 g g -1 or 0.5:10 to 1:10 g g -1 or 1:10 to 5:10 g g -1 or 1:10 to 4:10 g g -1 or 1:10 to 3:10 g g -1 or 1:10 to 2:10 g g -1 , and the ratio can be 1:1 or 1:2 or 1:3 or 1:4 or 1:5 or 1:6 or 1:7 or 1:8 or 1:9 or 1:10 g g -1 , more preferably, the ratio is 1:5 g g -1 .

[0110] In a preferred embodiment, the weight ratio of cannabinoid to amphiphilic carbohydrate is higher than 1:0.5, preferably 1:1 to 1:10, more preferably 1:5 to 1:3. This ratio is usually used in the case of GCPQ as the amphiphilic carbohydrate, and GCPQ has the above-mentioned preferred levels of palmitoylation and quaternization. Thus, in another aspect of the present invention, there is provided a composition comprising GCPQ and a cannabinoid, wherein the palmitoylation level of GCPQ is 11-31 mol%, the quaternization level is 8-17 mol%, and the weight ratio of GCPQ to cannabinoid is 1:1 to 1:10. Preferably, the composition is a pharmaceutical composition comprising GCPQ and a non-psychoactive cannabinoid and one or more pharmaceutically acceptable excipients.

[0111] The resulting nanoparticle composition can be converted into a nano-microparticle composition by further processing. For example, the nano-microparticle composition can be prepared by spray drying or freeze-drying the nanoparticle composition. The nano-microparticle composition can also be prepared by adding the nanoparticle composition to a powder to form granules. The nano-microparticle composition can be in the form of a dry powder.

[0112] The nano-microparticle composition is particularly suitable for reliably and selectively delivering large micron-sized particles to the top of the nasal cavity, causing the micron-sized particles to disintegrate into smaller particles and effectively penetrate the mucus to facilitate mucosal absorption.

[0113] The nanoparticles and nano-microparticle compositions according to the present invention are also capable of being stored for at least 30 days without a significant decrease in their stability and properties (such as particle size and morphology). The formulations of the present invention should also remain stable for a long time, as measured by the stability of the API (especially cannabinoids) over time. After storing the formulations of the present invention at 4 °C or 25 °C for 4 weeks, the recovery rate of the API is at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 99%. After storing the formulations of the present invention at 40 °C for 4 weeks, the recovery rate of the API is at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 99%. After storing the formulations of the present invention at 25 °C for 4 weeks, the recovery rate of the API is about 50% to about 99%, or about 50% to about 90%, or about 50% to about 80%, or about 50% to about 70%, or about 50% to about 60%, or about 60% to about 99%, or about 60% to about 90%, or about 60% to about 80%, or about 60% to about 70%, or about 70% to about 99%, or about 70% to about 90%, or about 70% to about 80%, or about 80% to about 99%, or about 80% to about 90%, or about 90% to about 99%. After storing the formulations of the present invention at 40 °C for 4 weeks, the recovery rate of the API is about 50% to about 99%, or about 50% to about 90%, or about 50% to about 80%, or about 50% to about 70%, or about 50% to about 60%, or about 60% to about 99%, or about 60% to about 90%, or about 60% to about 80%, or about 60% to about 70%, or about 70% to about 99%, or about 70% to about 90%, or about 70% to about 80%, or about 80% to about 99%, or about 80% to about 90%, or about 90% to about 99%.

[0114] The compositions of the present invention can be further formulated to form pharmaceutical compositions particularly suitable for nasal delivery.

[0115] Other ingredients that may be included in the pharmaceutical composition include: tonicity enhancer, preservative, solubilizer, non-toxic excipient, demulcent, chelating agent, pH regulator, co-solvent, and thickening agent.

[0116] The pharmaceutical composition can be formulated into a powder or a liquid dispersant. The delivery of the nasal spray can be achieved through a spraying device.

[0117] The dosage can be determined according to age, body weight, administration time, administration method, drug combination, severity of the clinical condition, or the actual condition of the patient receiving treatment, and other factors. Although the daily dosage may vary depending on the patient's condition and body weight, type of drug or active ingredient, and administration route, in the case of oral use, the daily dosage may be about 0.1 mg - 10.0 g / person / day, or 0.1 mg to 5.0 g / person / day, or 0.1 mg to 1.0 g / person / day, or 0.1 mg to 2.5 g / person / day, or 0.1 mg to 1.0 g / person / day, or 0.1 mg to 500 mg / person / day, or 0.5 mg to 5 g / person / day, or 0.5 mg to 2.5 g / person / day, or 0.5 mg to 1.0 g / person / day, or 0.5 mg to 0.5 g / person / day, or 0.5 mg to 100 mg / person / day, or 1 mg to 2.5 g / person / day, or 1 mg to 1.0 g / person / day, or 1 mg to 0.5 g / person / day, or 1 mg to 1.0 g / person / day, or 1 mg to 500 mg / person / day, or 10 mg to 2.5 g / person / day, or 10 mg to 1.0 g / person / day, or 10 mg to 0.5 g / person / day, or 10 mg to 0.1 g / person / day, or 10 mg to 50 mg / person / day, or 0.5 - 5000 mg / person / day, or 5 - 500 mg / person / day, or 10 - 250 mg / person / day, or 25 - 200 mg / person / day.

[0118] The present invention will be illustrated by the following examples.

[0119] Example

[0120] Example 1 - Synthesis of GCPQ-CBD Nanoparticles

[0121] Materials and Methods

[0122] CBD-GCPQ nanoparticles were synthesized at two different concentration ratios: 1:5 and 1:10 (CBD:GCPQ).

[0123] Cannabidiol (CBD > 99%; THC Pharm) was dissolved in methanol at a concentration of 1 mg mL -1 for both ratios. Meanwhile, GCPQ GC17.5 P 18 Q 13 Dissolve them separately in methanol at concentrations of 5 mg / mL -1 and 10 mg / mL -1 respectively. Then mix the solutions together as follows: Drop the CBD solution into the GCPQ solution while gently shaking, and then vortex for a few seconds. Incubate the mixture at 150 rpm for 1 h at room temperature. Next, evaporate the CBD-GCPQ preparation at 45 °C for 2 h using a rapid vacuum concentrator, and rehydrate the formed film in Milli-Q water at concentrations of 5 mg / mL -1 (1:5 ratio) and 10 mg / mL -1 (1:10 ratio). Figure 1 A schematic diagram of this method is provided.

[0124] Characterization method

[0125] The synthesized CBD-GCPQ nanoparticles were then characterized for colloidal stability and CBD content quantification.

[0126] Colloidal stability:

[0127] Characterization was performed using dynamic light scattering (DLS) technology. The samples were diluted 30-fold with Milli-Q water, and particle size, polydispersity, and zeta potential parameters were measured using a folded capillary zeta potential sample cell on a Malvern Zetasizer Nano S / N. The measurement methods for particle size and zeta potential included: Each sample was equilibrated for 30 s before data acquisition, and the measurements were repeated three times.

[0128] CBD quantification:

[0129] High-performance liquid chromatography (HPLC) was used to detect and quantify CBD in non-biological samples of the MET-CBD preparation, performed using an Agilent HPLC-UV system 1220 Infinity LC with a mobile phase of methanol:water 85:15% v / v. All samples submitted for HPLC were diluted 100-fold with methanol. Data analysis was performed using Agilent Chemstation software.

[0130] Results

[0131] Based on adding known concentrations of CBD in methanol, the HPLC results for quantifying CBD generated a linear calibration curve in the range of 3 to 100 μg / mL( Figure 2 A). Additionally, a distinct single CBD peak was formed in the CBD-GCPQ preparation at a retention time of approximately 4 minutes.

[0132] At 1 mg / mL-1 At the CBD equivalent concentration, the formulation showed good colloidal stability parameters: particle size less than 200 nm, low polydispersity (about 0.1) and a highly positive surface charge of about +50 mV( Figure 3 ). For in vivo applications, higher CBD concentrations are required, so attempts were made to use 5 - 10 mg mL -1 CBD equivalents in the CBD-GCPQ nanoparticle formulation. 10 mg mL -1 could not be achieved because the formulation could not be fully dispersed at such high concentrations and still had high viscosity and solid residues despite prolonged vigorous vortexing and sonication bath treatment. This was especially true for the formulation with a CBD:GCPQ ratio of 1:10 g g -1 because this formulation had a high level of GCPQ and thus a high solid content. The CBD-GCPQ formulation was successfully prepared at 5 mg mL -1 CBD in water, and for both CBD:GCPQ ratios (1:10 and 1:5 g g -1 ), colloidal stable nanoparticles were prepared with a unimodal particle population distribution and no signs of precipitation or aggregation. Compared to the CBD-GCPQ with a 1:10 g g -1 ratio, the CBD-GCPQ formulation with a 1:5 g g -1 ratio produced better particle size parameters, i.e., compared to 551 nm (CBD-GCPQ = 1:10 g g -1 ), the nanoparticles had a particle size of approximately 412 nm (CBD-GCPQ = 1:5 g g -1 ), and the polydispersity index (PDI) was slightly lower, at 0.14 (CBD-GCPQ = 1:5 g g -1 ), compared to 0.2 (CBD-GCPQ = 1:10 g g -1 ).

[0133] Example 2 - MET-CBD nano-micron particle powder

[0134] Materials and methods

[0135] According to the method of Example 1, CBD-GCPQ nanoparticles were prepared using CBD (60 mg) and GCPQ (300 mg) in a ratio of 1:5, and then the CBD-GCPQ nanoparticles were resuspended in 12 mL of Milli-Q water before spray drying. The nanoparticle dispersion was spray dried to obtain CBD-GCPQ nano-microparticles (Büchi micro spray dryer B290, Büchi Labortechnik AG, Switzerland), and the parameters were set as follows: inlet temperature = 180 °C, outlet temperature = 120 °C, aspirator % = 85%, pump % = 5%, ultrasonic controller = 1.8 (temperature < 50 °C).

[0136] Characterization methods

[0137] The synthesized CBD-GCPQ nanoparticles were then characterized for microparticle morphology and particle size analysis, colloidal stability, and quantification of CBD content in nanoparticles in abiotic and biological samples. Some spray-dried CBD-GCPQ powders were stored at room temperature for 30 days, while other samples were stored at 4 °C for 30 days. Samples were taken at different time points (0, 7, 14, and 30 days) for characterization.

[0138] Microparticle size:

[0139] The microparticle size of the spray-dried MET-CBD nano-microparticles was determined by laser scattering using a Malvern Mastersizer 3000. A (approx. 10 mg) powder sample was applied to the sample feed tray. Air was used as the dispersion medium for the microparticles entering the sample cell from the sample feed tray. The D 10 、D 50 and D 90 parameters were used to characterize the microparticle size distribution.

[0140] Microparticle morphology:

[0141] The morphology of the microparticles was determined by scanning electron microscopy (SEM) imaging. A double-sided carbon tape was placed on the SEM sample stage, and the CBD-GCPQ powder was spread over the surface of the tape, and the loose microparticles were removed with compressed air. The sample was covered with a 20 nm sputtered gold layer before measurement. SEM images of the sample were generated using a Phenom Pro Benchtop SEM.

[0142] Colloidal stability:

[0143] Samples were prepared and characterized for colloidal stability according to the method described in Example 1.

[0144] CBD quantification:

[0145] Prepare samples according to the method described in Example 1 and characterize them for the quantification of CBD in non-biological samples.

[0146] Results

[0147] SEM characterization showed that the spray-dried particles were mostly spherical, while the morphology of the individual CBD powder was more irregular. Table 1 below provides the average particle size of the particles (including SEM error values):

[0148] Table 1: Average particle size, D 10 , D 50 , D 90 and percentage of particles smaller than 10 μm (%)

[0149] <![CDATA[D 10 (μm)]]> <![CDATA[D 50 (μm)]]> <![CDATA[D 90 (μm)]]> % < 10 μm Average particle size 14.59±2.62 22.09±3.13 34.37±4.18 7.51±2.18

[0150] The spray-dried particles were rehydrated in water to a CBD concentration of 5.4 mg mL -1 (by HPLC analysis), generating nanoparticles with a particle size of approximately 500 nm, which exhibited a unimodal particle size distribution with low polydispersity (PDI ≤ 0.2). Compared with the nanoparticles before spray drying, the particle size of the rehydrated nanoparticles seemed to increase slightly. However, this was most likely attributed to the difference in the actually measured CBD concentration, which in turn led to different GCPQ contents in the two CBD-GCPQ formulations, i.e., 5.43 mg mL -1 for the formulation after spray drying, while the original formulation was 4.93 mg mL -1 , as Figure 4 shown. This may have caused a slight increase in particle size, as this is a common trend for concentrated formulations. The polydispersity before and after spray drying remained at approximately 0.19 ( Figure 4 ).

[0151] During storage at room temperature and 4 °C for 30 days, the spray-dried CBD-GCPQ micron particles maintained a spherical morphology at multiple test time points, namely day 0, day 7, day 14, and day 30. Tables 2 and 3 below provide all the particle size parameters of the spray-dried CBD-GCPQ powder stored at room temperature and 4 °C for 0, 7, 14, and 30 days, respectively.

[0152] Table 2: Particle size distribution of nano-micron particles stored at room temperature for 0, 7, 14, and 30 days

[0153] <![CDATA[D 10 (μm)]]> <![CDATA[D 50 (μm)]]> <![CDATA[D 90 (μm)]]> % < 10 μm Day 0 18.71±5.06 41.56±7.69 67.16±13.08 4.10±3.72 Day 7 11.94±2.82 28.48±7.98 57.99±20.88 13.58±5.06 Day 14 18.33±2.70 40.57±1.87 67.58±1.98 4.67±5.06 Day 30 12.13±2.49 22.41±5.43 42.56±14.14 13.04±5.06

[0154] Table 3: Particle size distribution of nano-micron particles stored at 4 °C for 0, 7, 14, and 30 days

[0155] <![CDATA[D 10 (μm)]]> <![CDATA[D 50 (μm)]]> <![CDATA[D 90 (μm)]]> % < 10 μm Day 0 18.71±5.06 41.56±7.69 67.16±13.08 4.10±3.72 Day 7 13.06±2.94 30.94±4.58 71.56±12.82 10.12±4.21 Day 14 21.85±4.14 33.07±6.03 49.64±7.97 3.29±3.28 Day 30 10.42±1.83 23.69±5.85 43.01±9.45 15.44±3.12

[0156] During storage at two different temperatures for 30 days, the median volume distribution (D 50 ) and the percentage of particles smaller than 10 mm (%) did not show statistically significant differences.

[0157] Then, the spray-dried CBD-GCPQ powder was rehydrated to 5 mg / mL CBD equivalent, and the colloidal stability and CBD concentration of the nanoparticles were analyzed and compared with the parameters on day 0. As Figure 5 shown, no statistically significant differences in particle size were observed over time. Similarly, no significant differences were observed in polydispersity and zeta potential at room temperature or 4 °C for up to 30 days ( Figure 6 A and 6B). No CBD degradation was observed in the CBD-GCPQ nanoparticle formulation at either temperature for up to 30 days ( Figure 6 C), although the refrigerated powder unexpectedly showed significantly higher CBD concentrations during this period (on days 14 and 30). In contrast, Mazzetti et al. reported in the Scientific Report of Nature Research (2020, 10:3697) that commercially available CBD oil-based e-liquid degraded by an average of 13% when exposed to light at room temperature for 30 days. Therefore, compared with the prior art, the present invention shows significant stability advantages by forming a stable and effective non-psychoactive cannabinoid - amphiphilic carbohydrate composition.

[0158] Example 3 - In Vivo Pharmacokinetic Study of Intranasal Administration of MET-CBD in Rats

[0159] Materials and Methods

[0160] Animals:

[0161] Male Sprague Dawley rats (Charles River, UK) were used, five rats per cage, and they were housed in an air-conditioned control unit (temperature 20 - 22 °C, relative humidity 50 - 60%), allowing free access to standard rodent feed and water. The lighting was controlled in a 12-hour cycle (lights on at 7:00 and lights off at 19:00). Before the experiment, the animals were acclimated to the environment for 7 days and to the experimental procedure room for 1 hour before the test. The body weight of the rats before dosing was 200 - 230 g, and the rats were grouped so that the average body weight of each experimental group was matched.

[0162] In Vivo Application and Intranasal Administration of the CBD-GCPQ Formulation:

[0163] Before administering the drug to rats, the CBD-GCPQ preparation prepared according to Example 2 was dispersed in Milli-Q water at a CBD concentration of 5 mg / mL. -1 The pH value of the preparation was measured and adjusted to 5.5 - 5.6 by adding NaOH. The rats were divided into 5 experimental groups with 6 rats in each group. Four of the groups were administered CBD-GCPQ and sacrificed at 4 different post-administration time points (10 min, 30 min, 1 h, 2 h), and one group was not administered. The CBD-GCPQ was administered intranasally using a Smiths Medical Portex fine pore polyethylene tube (inner diameter / outer diameter 0.28 / 0.61 mm, length 15 mm) connected to a 0.3 mL insulin syringe with a 30G needle at a dose of 2 mg / kg - 1 CBD equivalent). The animals were anesthetized by inhaling approximately 4% isoflurane for a few minutes before administration.

[0164] Tissue extraction and blood collection:

[0165] At the time points of 10 min, 30 min, 1 h, and 2 h, the rats after administration were sacrificed by carbon dioxide asphyxiation, and death was confirmed by cervical dislocation. Immediately after sacrifice, blood samples were collected by cardiac puncture and collected into microtubes containing K3EDTA anticoagulant. The blood samples were subsequently used for plasma separation.

[0166] The plasma was separated from the blood cells by centrifugation, the plasma was aspirated into 1.5 mL Eppendorf microtubes, and stored frozen at -50 °C until LC-MS analysis could be performed.

[0167] The brain was extracted and rapidly frozen in liquid nitrogen, and then stored frozen at -80 °C until LC-MS analysis could be performed. The brain homogenate and plasma were subsequently used as biological matrices for compound extraction and LC-MS analysis.

[0168] Characterization

[0169] The synthesized CBD-GCPQ nanoparticles were then characterized for the CBD content in biological samples (rat brain and plasma).

[0170] CBD quantification:

[0171] Quantify the CBD content in biological samples using liquid chromatography - mass spectrometry (LC - MS). Measure CBD and its internal standard deuterated CBD (CBD - d3, molecular weight = 318 Da) in brain and plasma matrices using an Agilent 6400 Series Triple Quad instrument. First, analyze abiotic samples containing known concentrations of the target compound CBD and the internal standard CBD - d3 added by LC - MS scanning. This process involves a three - step workflow: 1) perform a full tandem mass spectrometry (MS2) scan to identify the precursor ions of the two compounds; 2) perform a product ion scan, where the selected precursor ions are fragmented to form and identify product ions; and 3) based on the selected product ions, perform a multiple reaction monitoring (MRM) scan mode for the target compound CBD to quantify the target compound in the sample. The mobile phase of the binary pump is water:methanol, 95:5% v / v at 0 minutes and changes to 15:85% v / v at 2 minutes. The source parameters are set with a gas temperature of 300 - 350 °C.

[0172] Once quantifying analytes introduced into a biological matrix, the sensitivity of the LC - MS method usually decreases, and this phenomenon is commonly referred to as the "matrix effect". Therefore, before analyzing the brain and plasma in in - vivo studies, the MRM method was optimized by changing the mobile phase composition (i.e., with different polarities and organic solvents as well as isocratic and gradient compositions).

[0173] Results

[0174] CBD quantification in in - vivo pre - clinical studies:

[0175] The CBD quantification method showed that in abiotic samples, a distinct CBD peak was observed at a retention time of 1 minute by the isocratic method. In the biological matrix, a distinct peak was also observed at a retention time of 4.4 minutes by the gradient method.

[0176] The first MS2 scan identified the most abundant CBD precursor ion with a mass-to-charge ratio (m / z) of 315.2, and the m / z of the CBD-d3 ion was 318.2, which was consistent with the literature values provided by McRae, G et al. (Quantitative determination and validation of 17 cannabinoids in cannabis and hemp using liquid chromatography-tandem mass spectrometry. Analytical and Bioanalytical Chemistry, 412:7381-7393, 2020). Subsequently, this CBD precursor ion was selected for fragmentation in the second workflow to produce product ions. The product ion scan showed that the CBD precursor ion was completely fragmented into product ions. The m / z = 193 of the CBD product ion and the m / z = 196.3 of the CBD-d3 product ion were also confirmed by comparison with the study of McRae G et al. and were selected for the third workflow.

[0177] Based on the mass-to-charge ratios identified in the MS2 and product ion scans, a MRM method for the quantification of CBD and CBD-d3 was established, and calibration curves were generated in abiotic samples, showing very good linearity in the quantification range of 1 - 500 ng / mL, as Figure 7 shown. The distinct CBD peak in the MRM scan confirmed the accuracy of the LC-MS quantification method and the high sensitivity of the method (the lower limit of quantification (LLOQ) was at least 1 ng / mL).

[0178] Calibration curves were generated in brain tissue and plasma matrices, as Figure 8 shown, showing good linearity in the quantification ranges of 1 - 100 ng / mL (brain, R 2 = 0.999) and 3 - 200 ng / mL (plasma, R 2 = 0.995), respectively. The LC-MS method showed high sensitivity in CBD quantification, with LLOQs of 1 ng / mL and 3 ng / mL in brain and plasma matrices, respectively.

[0179] Colloidal stability, pH value, and particle size distribution:

[0180] Before dosing the rats, for 5 mg mL -1The colloidal stability and pH value of the nanoparticles were characterized. The nanoparticles formed a monodisperse (PDI < 0.1) population of nanoparticles with a particle size of less than 300 nm and a pH value of 4.5 - 4.8. This pH value is lower than the physiological pH value in the nasal cavity - according to the research report by England et al., the physiological pH in the nasal cavity is 5.5 - 5.6 (Nasal pH measurement: a reliable and repeatable parameter. Clin Otolaryngol Allied Sci, 24(1): 67 - 8, 1999). Since this may cause discomfort in animals and adverse reactions such as sneezing, the pH value was adjusted to this range before drug administration. The increase in pH value had a minimal effect on the colloidal stability of the nanoparticles; the particle size of the nanoparticles only slightly increased to about 320 nm, and the polydispersity index remained at about 0.1. Figure 9 The particle size distribution and polydispersity before and after pH adjustment were compared.

[0181] In vivo pharmacokinetic study of intranasal administration:

[0182] Overall, the intranasal administration of CBD - GCPQ was very well tolerated in the animals administered. No sneezing, distress, or adverse reactions were reported; the rats seemed calm and relaxed. As reported by Galaj et al. (Possible Receptor Mechanisms Underlying Cannabidiol Effects on Addictive - like Behaviors in Experimental Animals. Int. J. Mol. Sci., 22(1), 134, 2021), CBD can bind to certain receptors in the brain, release serotonin, and thus produce a sense of well - being; while THC has a psychostimulant effect and may induce addictive behaviors. Therefore, the observed calm behavior of the rats after drug administration indicates that CBD reached the brain quite rapidly.

[0183] Figure 10 Shows administration of 2 mg kg -1CBD levels in the brain, olfactory bulb, and plasma of test rats at 0 minutes (untreated control group), 10 minutes, 30 minutes, 60 minutes, and 120 minutes. These results confirm that significant levels of CBD are present in the brain, olfactory bulb, and plasma within 10 minutes after administration. For all curves, the maximum concentration is reached at 30 minutes, and the concentration gradually decreases after 60 minutes. Even 120 minutes after administration, a certain level of CBD remains in the brain and olfactory bulb. In plasma, the maximum concentration of CBD is 40-fold lower than in the brain, indicating very low systemic exposure after intranasal administration. These results are superior to the CBD levels reported in the literature. For example, Hozek et al. (Pharmacokinetic and behavioural profile of THC, CBD, and THC+CBD combination after pulmonary, oral, and subcutaneous administration in rats and confirmation of conversion in vivo of CBD to THC. European Neuropsychopharmacology, 27, 1223-1237, 2017) administered by oral inhalation at 10 mg kg -1 and 20 mg kg -1 and the CBD levels in the brain only reached approximately 200-300 ng g -1 , significantly lower than the 2,400 ng g -1 reported in the present invention (as shown in Figure 10 A). Therefore, the intranasal administration method of the present invention achieves a CBD concentration 10 times higher than that of the prior art, even when using a much lower dose (2 mg kg -1 ). Therefore, the present invention allows the use of a lower clinical dose of the amphiphilic carbohydrate-non-psychoactive cannabinoid composition.

[0184] Example 4: Preparation and Characterization of Further Compositions

[0185] The aim of this example was to prepare compositions comprising GCPQ with different amounts of palmitoylation (d unit in formula I) and quaternization (b unit). The effects of different cannabinoid to GCPQ ratios were also tested.

[0186] Materials and Methods

[0187] The CBD (THC Pharm, UK) and GCPQ formulations were prepared by thin-film evaporation. Several GCPQ polymers with different molar percentages of palmitoylation (%P) and molar percentages of quaternary ammonium groups (Q%) were studied, where the levels of palmitoylation and quaternary ammonium groups were expressed by the following polymer designations: GCP11Q11; GCP19Q8; GCP20Q13; GCP20Q17; GCP31Q13. In these designations, for example, GCP11Q11 refers to a polymer with 11 mol% palmitoylation and 11 mol% quaternary ammonium groups.

[0188] In a round-bottom flask, CBD and MET were dissolved in MeOH at CBD to MET ratios of 1:0.5, 1:1, 1:3, 1:5, and 1:10, with a final CBD concentration of 1 mg / mL. The MeOH mixture was stirred at room temperature for 1 h. Subsequently, a thin film was obtained by rotary evaporation under vacuum in a 45 °C water bath, and the film was rehydrated with Milli-Q water to a final CBD concentration of 5 mg / mL. -1 The dispersion was sonicated in an ultrasonic bath at 30 °C for 10 minutes.

[0189] Dynamic light scattering and Zeta potential

[0190] Dynamic light scattering (DLS) and zeta (ζ) potential measurements were performed using a Zeta-sizer Ultra (Malvern Instruments Ltd). The dispersion was diluted 30-fold with Milli-Q water and measured after equilibration at 25 °C for 120 s. The measurements were repeated three times. The software used CONTIN analysis to obtain the intensity distribution and cumulative analysis to obtain the hydrodynamic diameter (DH) and polydispersity index (PI).

[0191] Quantification of CBD content by high-performance liquid chromatography (HPLC)

[0192] The CBD content in the MET-CBD formulations was quantified using HPLC with the following settings:

[0193] HPLC method:

[0194] Agilent HPLC-UV system 1260 Infinity II

[0195] Column: Agilent Eclipse Plus C-18 4.6 mm × 150 mm, 3.5 μm

[0196] Mobile phase: methanol: water (85:15%, v / v)

[0197] Detection wavelength: 220 nm

[0198] Flow rate: 1.0 mL min -1

[0199] Injection volume: 10 μL

[0200] Run time: 6 min

[0201] Retention time: approximately 4 min

[0202] A linear calibration curve (R 2 = 0.9996) was generated in the range of CBD concentration from 1 to 150 μg / mL in methanol. A distinct single CBD peak was found in the MET-CBD formulation, with the same retention time as that of the individual CBD (approximately 4 min). All samples submitted for HPLC were diluted 100-fold with methanol. Data analysis was performed using Agilent Chemstation software.

[0203] Results

[0204] The prepared formulations were characterized in terms of particle size, polydispersity index (PI), ζ-potential, and CBD content (%). The stability of these formulations depends on the type of polymer used (in terms of %P and %Q) and the ratio of CBD to the polymer. Their stability was tracked over time, up to 8 hours or until the formation of a suspension / precipitate. In all cases, precipitation was observed after 24 hours.

[0205] Table 4: Summary of results of CBD:GCP11Q11 over time (Z-average, PI, ζ-potential, and CBD content)

[0206] CBD:GCP11Q11

[0207]

[0208]

[0209] Table 5: Summary of results of CBD:GCP19Q8 over time (Z-average, PI, ζ-potential, and CBD content) (MW 12 kDa)

[0210] CBD:GCP19Q8

[0211]

[0212] Table 6: Summary of results of CBD:GCP20Q13 over time (Z-average, PI, ζ-potential, and CBD content)

[0213] CBD:GCP20Q13

[0214]

[0215]

[0216] 1 Multi-modal particle size distribution

[0217] Table 7: Summary of the results of CBD:GCP20Q17 over time (Z-average, PI, ζ potential, and CBD content) (MW = 11.9 kDa)

[0218] CBD:GCP20Q17

[0219]

[0220]

[0221] Table 8: Summary of the results of CBD:GCP31Q13 over time (Z-average, PI, ζ potential, and CBD content)

[0222] CBD:GCP31Q13

[0223]

[0224]

[0225] From the above results, it can be seen that GCPQ polymers with a palmitoylation molar percentage (%P) less than or equal to 31%, typically 11% to 20%, are preferred. In addition, GCPQ polymers with a quaternary ammonium group molar percentage (%Q) less than or equal to 17%, typically 8 to 17%, are preferred.

[0226] The above results also show that the preferred CBD:GCPQ ratio is higher than 1:0.5, and ideally 1:1 to 1:10.

Claims

1. A method of treatment, comprising administering to a human or an animal a composition comprising a cannabinoid and an amphiphilic carbohydrate, wherein the composition is administered intranasally to the human or animal body.

2. A composition comprising a cannabinoid and an amphiphilic carbohydrate for use in a method of treating a human or an animal by intranasal administration.

3. The method or composition for use according to claim 1 or 2, wherein the treatment is for a central nervous system disease, preferably for the treatment of epilepsy.

4. The method or composition for use according to any one of claims 1 to 3, wherein the treatment is for the treatment of pain, anxiety or an autoimmune disease.

5. The method or composition for use according to any of the preceding claims, wherein the composition is in the form of nanoparticles.

6. The method or composition for use according to claim 5, wherein the nanoparticles are processed to form nano-microparticles.

7. The method or composition for use according to claim 6, wherein the composition is prepared by a method of forming nano-microparticles selected from spray drying, freeze drying or adding nanoparticles to a powder to form particles, preferably wherein the method is spray drying.

8. The method or composition for use according to claim 6 or 7, wherein the average particle size of the nanoparticles in the nano-microparticle composition is less than 1000 nm, preferably wherein the particle size of the nanoparticles is less than 500 nm.

9. The method according to any one of claims 6 to 8 or the composition for said use, wherein the median volume distribution D of the micron particles in the nano-micron particle composition 50 is from 5 to 30 μm, preferably from 10 to 25 μm.

10. The method or composition for use according to claim 9, wherein less than 10% of the particle sizes are below 10 μm.

11. The method or composition for use according to any one of claims 6 to 10, wherein the polydispersity of the nanoparticles in the nano-microparticles is less than 0.5, preferably less than 0.2, more preferably less than 0.

1.

12. The method or composition for use according to any one of claims 6 to 11, wherein the nanoparticles in the nano-microparticles are in a colloidally stable formulation.

13. The method or composition for use according to any one of claims 6 to 12, wherein the nano-microparticles are in dry powder form.

14. The method or composition for use according to claim 13, wherein the nano-microparticles are compatible with a nasal spray device.

15. The method or composition for use according to any one of claims 6 to 14, wherein the nano-microparticles have a spherical, hollow or irregular morphology.

16. The method according to any one of the preceding claims or the composition for said use, wherein the ratio of cannabinoid to amphiphilic carbohydrate is from 0.5:10 to 5:10 g / g -1 .

17. The method according to claim 16 or the composition for the use, wherein the ratio of cannabinoid to amphiphilic carbohydrate is from 1:10 to 3:10 g / g -1 , preferably wherein said ratio is about 1:5 g / g -1 .

18. The method or composition for use according to any of the preceding claims, wherein the cannabinoid is delivered to the brain of the human or animal body.

19. The method or composition for use according to any of the preceding claims, wherein the cannabinoid is a non-psychoactive cannabinoid, preferably cannabidiol.

20. The method or composition for use according to any of the preceding claims, wherein the amphiphilic carbohydrate is a chitosan derivative.

21. The method according to any one of the preceding claims or the composition for the use, wherein the amphiphilic carbohydrate comprises the following general formula: where a + b + c + d = 1.000, and a is between 0.01 and 0.970, b is between 0.01 and 0.990, c is between 0.0001 and 0.970, and d is between 0.01 and 0.990; and wherein X is a hydrophobic group; R 1 、R 2 and R 3 are each independently selected from substituted or unsubstituted alkyl groups; R 4 、R 5 、R 6 and R 10 are each independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted ether group, or substituted or unsubstituted alkenyl; R 7 may be present or absent. When R 7 is present, it is an unsubstituted or substituted alkyl group, an unsubstituted or substituted amino group, or a substituted or unsubstituted amide group; R 8 and R 9 are each independently selected from hydrogen, and substituted or unsubstituted alkyl, substituted or unsubstituted ether group, or substituted or unsubstituted alkenyl; or a salt thereof.

22. The method according to claim 21 or the composition for the use, wherein the amphiphilic carbohydrate is quaternary ammonium palmitoyl glycol chitosan (GCPQ).

23. A pharmaceutical composition suitable for intranasal administration, comprising an amphiphilic carbohydrate, a non-psychoactive cannabinoid, and one or more pharmaceutically acceptable excipients.

24. The pharmaceutical composition according to claim 23, wherein the cannabinoid is selected from cannabidiol (CBD), cannabinol (CBG), cannabidiolic acid (CBDA), cannabidivarin (CBDV), cannabichromene (CBC), and combinations thereof.

25. The pharmaceutical composition according to claim 24, wherein the cannabinoid is cannabidiol (CBD).

26. The pharmaceutical composition according to any one of claims 23 to 25, wherein the excipient is selected from osmotic promoters, preservatives, solubilizers, non-toxic excipients, emollients, chelating agents, pH regulators, co-solvents, thickeners, and combinations thereof.

27. The pharmaceutical composition according to any one of claims 23 to 26, wherein the amphiphilic carbohydrate is GCPQ.

28. The pharmaceutical composition according to any one of claims 23 to 27, wherein the composition is in the form of nanoparticles.

29. The pharmaceutical composition according to claim 28, wherein the nanoparticles are processed to form nano-microparticles.

30. The pharmaceutical composition according to claim 28 or 29, wherein the average particle size of the nanoparticles in the nano-microparticle composition is less than 1000 nm, preferably wherein the particle size of the nanoparticles is less than 500 nm.

31. The pharmaceutical composition according to any one of claims 28 to 30, wherein the median volume distribution D 50 of the micron-sized particles in the nano-micron particle composition is 5 to 30 μm, preferably 10 to 25 μm.

32. The pharmaceutical composition according to claim 31, wherein less than 10% of the particle size is below 10 μm.

33. The pharmaceutical composition according to any one of claims 29 to 32, wherein the polydispersity of the nanoparticles in the nano-microparticles is less than 0.5, preferably less than 0.2, more preferably less than 0.

1.

34. The pharmaceutical composition according to any one of claims 29 to 33, wherein the nanoparticles in the nano-microparticles are in a colloidally stable formulation.

35. The pharmaceutical composition according to any one of claims 29 to 33, wherein the nano-microparticles are in dry powder form.

36. The pharmaceutical composition according to claim 35, wherein the nano-microparticles are compatible with a nasal spray device.

37. The pharmaceutical composition according to any one of claims 29 to 36, wherein the nano-microparticles have a spherical morphology.

38. The pharmaceutical composition according to any one of claims 23 to 37, wherein the ratio of cannabinoid to amphiphilic carbohydrate is from 0.5:10 to 5:10 g / g -1 .

39. The pharmaceutical composition according to claim 38, wherein the ratio of cannabinoid to amphiphilic carbohydrate is from 1:10 to 3:10 g / g -1 , preferably wherein said ratio is about 1:5 g / g -1 .

40. The pharmaceutical composition according to any one of claims 23 to 39, wherein cannabidiol is stable for at least 4 weeks at 4°C.

41. The pharmaceutical composition according to any one of claims 23 to 40, wherein cannabidiol is stable for at least 4 weeks at 25 °C.

42. The pharmaceutical composition according to any one of claims 23 to 41, wherein the recovery rate of cannabidiol is at least 99% after storage at 4 °C for 4 weeks.

43. A pharmaceutical composition comprising GCPQ, cannabinoids, and one or more pharmaceutically acceptable excipients.

44. The pharmaceutical composition according to claim 43, wherein the palmitoylation level of GCPQ is 11 - 31 mole %.

45. The pharmaceutical composition according to claim 43 or 44, wherein the quaternization level of GCPQ is 8 - 17 mole %.

46. The pharmaceutical composition according to any one of claims 43 to 45, wherein the ratio of GCPQ to cannabinoids is from 1:1 to 1:

10.

47. The pharmaceutical composition according to claim 46, wherein the cannabinoids are selected from cannabidiol (CBD), cannabinol (CBG), cannabidiolic acid (CBDA), cannabidivarin (CBDV), cannabichromene (CBC), and combinations thereof.

48. The pharmaceutical composition according to claim 47, wherein the cannabinoid is cannabidiol (CBD).

49. A composition comprising GCPQ and cannabinoids, wherein the palmitoylation level of GCPQ is 11 - 31 mole %, the quaternization level of GCPQ is 8 - 17 mole %, and the ratio of GCPQ to cannabinoids is from 1:1 to 1:10.

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