Nasal spray composition and use thereof

Abstract

The present invention relates to a nasal spray composition comprising CBD and one or more permeation enhancers that provide good sprayability, CBD solubility, composition stability, and enhanced bioavailability of the composition, so that the composition discloses a good spray plume during application. The present invention further discloses the use of such composition in the manufacture of a medicament for aiding in the treatment of anxiety, depression and chronic pain.

Description

[0001] "Composition of Nasal Spray and its Use"

[0002] FIELD OF APPLICATION

[0003]

[0001] The present invention discloses a nasal spray composition comprising cannabidiol (CBD) and one or more permeation enhancers which, when combined with each other and different organic solvents, even at high concentrations of water in the mixture, provide good sprayability, CBD solubility, bioavailability and stability to the composition, such that the new composition exhibits good plume during application. The present invention further discloses the use of such composition in the manufacture of a medicine to aid in the treatment of anxiety, depression and chronic pain.

[0004] DESCRIPTION OF THE STATE OF THE ART

[0005]

[0002] Cannabis sativa is a plant that contains more than 400 compounds, including 66 phytocannabinoids. Delta-9-Tetrahydrocannabinol (A9-THC) is the main compound responsible for the hallucinogenic effects, while cannabidiol (CBD) is the second most abundant compound in the plant.

[0006]

[0003] Cannabis is a genus of flowering plants in the family Cannabaceae, where one or more species are recognized, depending on the taxonomic system employed (Polio A, Cannabis Cannabinoid Res. 2016; 1(1): 234-238). Currently, at least three species of the genus are recognized: Cannabis sativa, Cannabis indica, and Cannabis ruderalis. The long coexistence between humanity and the domestication of the plant has led to the use of this botanical genus in a variety of applications, including the use of hemp fibers, edibles, and psychoactive substances for medicinal and recreational purposes. The usefulness of Cannabis is being enhanced by biotechnology for the use of bioderivatives in the manufacture of bioplastics, antibacterial agents, phytochemicals, among others.

[0007]

[0004] Among plants of the genus Cannabis, Cannabis sativa produces a unique class of secondary metabolites called cannabinoids. Originally, the term cannabinoid represented a range of substances derived from Cannabis sativa, but this concept has been expanded to encompass all classes of substances, both natural and synthetic, that have pharmacological activity on cannabinoid receptors present in the human nervous system. A total of 66 phytocannabinoids have already been identified, most belonging to subclasses of the following structural classes: cannabigerol (CBG), cannabichromene (CBC), cannabidiol (CBD), A9-THC, A8-THC, cannabicyclol (CBL), cannabiesolin (CBE), cannabinol (CBN), cannabinodiol (CBDL), and cannabitriol (CBTL) (grotenhermen F, 2003). Much of the understanding of cannabinoid chemical structures, as well as evidence of their therapeutic potential, was described in the 1960s.

[0008]

[0005] As a highlight and second most abundant metabolite, cannabidiol (CBD) is a phenolic compound that can make up around 40% of plant extracts. With better acceptance as a medicinal substance, CBD is widely recognized for its ability to modulate the central nervous system through the endocannabinoid system. When administered to the human body, CBD establishes interactions with the neurotransmitter system, such as CB1, CB2, adenosine, and serotonin receptors. The CB1 receptor is found mainly in the central nervous system, such as the brain and spinal cord. It plays a fundamental role in regulating various brain functions and is associated with the psychotropic effects of cannabinoid compounds, such as alterations in pain perception and feelings of euphoria.On the other hand, the CB2 receptor is predominantly located in immune system cells and other peripheral tissues, and plays an important role in the immune response, while also being related to the modulation of chronic pain. This complex molecular interaction results in beneficial effects related to anxiety, inflammation, cognition, motor control, pain perception, nausea, and appetite.

[0009]

[0006] The best-known mechanism of action of CBD is the non-competitive antagonistic effect on CB1 and CB2 receptors. Cannabidiol limpes the activation of these receptors and thereby inhibits the inflammatory response and pain transmission. Furthermore, studies on the efficacy of CBD have shown positive results in the treatment of epilepsy, anxiety, Parkinson's disease, Alzheimer's disease, and pain.

[0010]

[0007] In 2018, the Food and Drug Administration (FDA) in the United States approved the therapeutic use of CBD as a pure, pharmaceutical-grade extract derived from a plant source, known as Epidiolex. This approval was granted specifically for the treatment of two severe forms of epilepsy: Lennox-Gastaut syndrome and Dravet syndrome.

[0011]

[0008] In clinical trials and research studies, CBD has been administered orally, either in capsule form or dissolved in oil. However, CBD undergoes a significant first-pass effect in the liver, meaning the compound is metabolized by the liver before reaching systemic circulation. These characteristics significantly reduce its bioavailability by more than 30% when used orally, making nasal administration a more advantageous option for this type of composition.

[0012]

[0009] The nasal route has been gaining attention as an alternative route for systemic drug delivery, especially drugs that undergo pre-systemic metabolism, with parts being metabolized, for example, by the liver, before reaching the bloodstream. Furthermore, the nasal route has advantageous characteristics such as a large surface area, the presence of a porous endothelial membrane, high blood flow, and ease of access. The epithelium with microvilli, vascularization, and porous basement membrane facilitates the absorption of drugs into the systemic circulation, especially lipophilic drugs. This allows for reduced doses, decreased adverse effects, and improved adherence to treatment.

[0013]

[0010] It is possible to didactically divide the respiratory tract into upper and lower. At rest, a human performs inspiration and expiration cycles between 12 and 15 times per minute, moving about 500 ml of air in each cycle (about 6-8 L / min) (COLOMBO, 2013). The respiratory tract has a large surface area that includes around 2.5 cm 2 area in the trachea and 11,800 cm 2 area in the alveoli, which is attractive as a therapeutic route (PAUDEL et al., 2010).

[0014]

[0011] Accumulated pharmaceutical knowledge reveals that the inhalation of an active substance must be in heterodispersed particles to reach the entire respiratory tract. Particles around 6 µm accumulate in the nasopharyngeal region, while particles < 3 µm reach the alveolar region (MAITI; BIDINGER, 1981). The efficiency in delivering active substances to the respiratory tract also involves other variables, such as: inspired volume, airflow, and cycle time. Particles can deliver the substance by inertial impact or sedimentation in the upper airways, and by Brownian diffusion in the bronchioles and alveoli. Humidity, temperature, and airway geometry also add complexity to the therapeutic strategy. The physicochemical nature of the particles can contribute to post-inhalation modifications, e.g.: hypertonic aqueous particles can increase in size with airway moisture; hygroscopic particles can merge and increase in size along the pathway; hypotonic particles can lose volume and decrease in size (SATALOFF; JOHNS; KOST, 2019).

[0015]

[0012] The pulmonary epithelium has a mucus layer that varies between 1-10 µm in thickness, and the alveolar surface has a thin layer of surfactants (0.1 - 0.2 µm thick). This organization directly influences the absorption dynamics in the respiratory tract. Metabolism is not as active as in the liver, and absorption by the alveoli is rapid. Solid particles can persist for 70 to 700 days in the lungs of rats and humans, respectively. Non-solid particles < 3 µm are phagocytosed in less than 12 h. Molecules < 6 nm can pass through the junction of blood capillary cells, and larger proteins end up being captured by phagocytic cells (COLOMBO, 2013).

[0013] The respiratory tract mucosa can move mucus at about 5 mm / min, promoting the elimination of foreign bodies in about 15 minutes. Active molecules can cross the mucosal surface by transcellular or paracellular transport.In general, lipophilic substances cross via transcellular pathways and polar substances via paracellular pathways. Paracellular transport has a size limitation due to intercellular junctions (~10 nm), creating a size cutoff around 1 kDa. These specifications directly affect the bioavailability of substances. Polar molecules have low bioavailability via the nasal route, e.g., <10% for morphine and <1% for insulin. However, lipophilic substances can reach 100% bioavailability (PAUDEL et al., 2010). Therefore, developing an inhalable pharmaceutical formulation is a challenge requiring multiplex strategies.

[0016]

[0014] There are several therapeutic options that utilize the nasal route. Many are already on the market, and some technological options are still in the testing phase. The strategic decision for the presentation and pharmaceutical technology of each drug or related product available on the market involves more than just the formulation. It is possible to find products based on the device, the composition, the dosage, and a mixture of these three. There are complex devices, propelled by propellants; others with intelligent mechanisms for dosimetry; others simpler with spray jets, and some that combine these specifications (SATALOFF; JOHNS; KOST, 2019).

[0017]

[0015] In this sense, document BR 11 2019 006106 1 provides compositions, preferably in the form of thermodynamically stable microemulsions, which can be administered intranasally, loaded with at least one cannabinoid comprising at least one oil, at least one hydrophilic surfactant, at least one co-surfactant, and / or a co-solvent and being devoid of water. One aspect of the present invention is to provide compositions that improve the solubilization of cannabinoids that exhibit low selectivity and poor bioavailability, impacting their therapeutic effects, without the use of nanotechnology, which implies altering the toxicity response of the cannabidiol carried in this type of composition.Another aspect of the present invention is to provide compositions characterized by generating an aerosol plume upon application, with substantially uniform and stable droplet size, typically on the micrometer scale, allowing for greater stability in droplet size so as not to impair the release of the cannabinoid. The composition is intended for the treatment of ailments including disorders associated with pain (as an analgesic), anxiety (as an anxiolytic), psychosis (as an antipsychotic), seizures (antiepileptic or antispasmodic), sleep (as an anti-insomnia treatment), post-traumatic stress disorder, among others.

[0018]

[0016] Document BR 11 2018 075073-5 provides high-viscosity or semi-solid compositions containing cannabinoids for nasal application, for the treatment of diseases or relief of symptoms such as epilepsy, pain, anxiety, spasticity, depression, among others. The document provides a nasal application composition, avoiding oral administration due to the low solubility of CBD in water, hindering absorption by the gastrointestinal system and impairing pharmacokinetics, in addition to the low bioavailability that the component presents in oleic-based compositions. The appearance of the aerosol plume formed in these compositions is not explored in this document.

[0019]

[0017] Document BR 112021024423-9 discloses thermogelling cannabinoid compositions, as well as their preparation method, suitable for transmucosal administration of the cannabinoid. The document provides compositions that are liquid at room temperature and transform into a gel when their temperature is increased to around body temperature. The document also refers to compositions that have a longer duration of intimate contact with mucous surfaces, intensifying the therapeutic effects of the active ingredient. The composition comprises an emulsion containing a cannabinoid and a viscosity modifier that ensure a liquid form at room temperature and a solid or semi-solid form around body temperature.

[0020]

[0018] The state of the art includes compositions containing CBD in pharmaceutical forms such as nasal sprays, microemulsions, hydrogels, viscous oil bases, aqueous bases, among others. However, the use of an aqueous base compromises the stability of the composition, since CBD is a lipophilic molecule that has low solubility in aqueous media. Additionally, it undergoes accelerated degradation in solvent systems with a high dielectric constant.

[0021]

[0019] On the other hand, existing oil-based compositions have impaired bioavailability when administered orally, given that CBD undergoes a significant first-pass effect in the liver.

[0022]

[0020] Moreover, prior art compositions generally have high viscosity, which prevents them from being administered via a common nasal device and from providing good spray with low weight variation in each application and adequate aerosol plume formation.

[0023]

[0021] Previous tests with compositions based on self-emulsifying drug delivery systems (SEDDS), a well-established strategy to improve the absorption and bioavailability of pharmaceutical compositions, have shown that, due to the high viscosity of some components of these systems, such as the surfactant Tween 80 and lipophilic vehicles with viscosity greater than 200 cP, an adequate plume is not formed when applied as a nasal spray. This makes their use as an alternative to low-bioavailability oral administration unfeasible.

[0024]

[0022] BENJAMIN and FENG, 2022 demonstrated that computational fluid dynamics can simulate nasal drug administration to (1) examine the effects on drug deposition when various external magnetic fields are applied to charged particles, (2) comprehensively study the effects of multiple parameters (i.e., aerodynamic particle diameter; magnitude of injection velocity, angle and position; magnetic force and direction), and (3) determine how to achieve optimal delivery efficiency to the olfactory epithelium. Reinforcing the importance of physicochemical parameters for greater efficiency of nasal spray-based medications, such as, for example, an ideal cone with up to a 30° angle at the actuation point. Complemented by SEIFELNASR et al., 2023, who demonstrated the best nasal delivery procedure.

[0025]

[0023] BR 102019025446-7 A2 demonstrates that the pharmaceutical composition in the form of a nasal spray, balanced in viscosity and density, when administered through a specific actuator, produces a particular and constant plume spray pattern, with a Gaussian droplet size distribution. This ensures greater efficacy in local topical nasal treatment without exceeding the nasopharyngeal region. In this way, the Physical Properties of the Formulation and a peculiar Spray Applicator Pump ensure that the product has a constant Plume Spray Pattern, and that it does not exceed the Larynx or Pharynx. Ensuring a localized topical effect.

[0026]

[0024] In this sense, the state of the art would benefit from the invention of a nasal-administered drug, composed of CBD, with low viscosity, high stability, solubility and bioavailability, and good sprayability in simple packaging devices, to be administered in the treatment of diseases such as anxiety, depression and chronic pain.

[0027] OBJECTIVES OF THE INVENTION

[0028]

[0025] One of the objectives of the present invention is to provide a nasal spray composition containing CBD and one or more permeation enhancers, in low concentration, allowing, with the use of lipophilic solvent mixtures, the obtaining of a composition with low viscosity, high stability, and consequently, good spraying and adequate plume formation.

[0029]

[0026] Another objective of the present invention is to provide a composition comprising CBD and permeation enhancers which, when combined with each other, exhibit a synergistic effect of increasing stability and providing good spreading capacity and solubility of CBD and, when combined with a lipophilic solvent, cause an increase in the bioavailability of the composition, also enabling the dilution of the lipophilic base mixture with water or saline solution, maintaining the stability and spreading capacity of the compositions obtained without obtaining a nanostructured system.

[0030]

[0027] Another objective of the present invention is the use of said composition in the manufacture of a medicament to assist in the treatment of anxiety, depression and chronic pain in the form of a nasal solution with increased bioavailability for cannabidiol.

[0031] SUMMARY OF THE INVENTION

[0032]

[0028] In order to achieve the aforementioned objectives, the present invention provides a nasal spray composition in the form of a pharmaceutical solution, comprising CBD, one or more permeation enhancers and one or more organic solvents, wherein the one or more permeation enhancers consist of lipophilic compounds which, combined, exhibit a synergistic effect providing good solubility, bioavailability of CBD and stability to said composition, and wherein a solvent combined with one or more permeation enhancers unusual for intranasal compositions, exhibit a synergistic effect providing the bioavailability of said composition and good spraying capacity.

[0033]

[0029] This pharmaceutical composition in solution form can be further diluted with water or saline solution between 0.1 and 1% w / v, while still remaining a clear solution, ensuring the stability, solubility and bioavailability of cannabidiol without the formation of nanostructured systems, microemulsions, hydrogels or other types of modified-release compositions.

[0034]

[0030] The present invention further discloses the use of a nasal spray composition in the preparation of a medicament that assists in the treatment of anxiety, depression and chronic pain.

[0035] BRIEF DESCRIPTION OF THE DRAWINGS

[0036]

[0031] The subject matter of this invention will become fully clear in its technical aspects from the detailed description that will be made based on the figures below, in which:

[0037] Fig. 1: The figure presents the analysis of the plume formed during the activation of the device with composition FP2.

[0038] Fig. 2: The figure presents the analysis of the plume formed during the activation of the device with composition FP3.

[0039] Fig. 3: The figure shows the weight variation in grams for each actuation of the device with the FP2 composition. The graph shows the number of actuations on the horizontal axis and the weight in grams of the expelled product on the vertical axis.

[0040] Fig. 4: The figure shows the weight variation in grams for each actuation of the device with composition FP3. The graph shows the number of actuations on the horizontal axis and the weight in grams of the expelled product on the vertical axis.

[0041] Fig. 5: The figure shows the variation in weight in grams for each activation of the device with the placebo (PEG 400 solution). The graph shows the number of activations on the horizontal axis and the weight in grams of the expelled product on the vertical axis.

[0042] Fig. 6: the figure shows the weight variation in grams for each activation of the device with "composition a"; Fig. 7: the figure shows the weight variation for the ethanol / PEG 200 1:1 mixture.

[0043] Fig. 8: The figure presents the in vitro cell viability assay according to ISO 10993 / c5 / AC. Cultured cells are exposed to the samples of interest. After incubation, dead cells are quantified and the graph shows the percentage of live cells in each condition. Each bar represents the average of triplicates (n=3). Statistical analysis confirms significance with p < 0.05 for CBD in placebo solution and in the FP3 composition (5 pg / mL) when compared to controls.

[0044] Fig. 9 shows the Dynamic Light Scattering (DLS) analysis of the spray obtained by diluting CBD at 5 pg / mL in a 0.9% w / v saline solution, "composition a". The analysis results reveal that the composition does not contain particles within the analyzed size range.

[0045] Fig. 10: the figure presents the analysis of the plume formed during activation of the device with “composition c”.

[0046] Fig. 11: the figure presents the analysis of the plume formed during the activation of the device with “composition d”.

[0047] DETAILED DESCRIPTION OF THE INVENTION

[0048]

[0032] In order to achieve the aforementioned objectives, the present invention provides a nasal spray composition in solution form, comprising CBD, using pharmaceutically acceptable non-aqueous solvents combined with lipophilic permeation enhancers, so as to provide a composition with improved sprayability, improved CBD solubility, greater stability and enhanced bioavailability by using a combination of two permeation enhancers at low concentrations, in this case, ethoxydiglycol (transcutol) and caprylocaproyl polyoxylglycerides 8 (labrasol).

[0033] In general, the combination of permeation enhancers and organic solvents in pharmaceutical compositions leads to obtaining solutions or suspensions of high viscosity.This high viscosity makes it difficult to spray them in the form of a nasal spray, for example, and also determines a greater difficulty in releasing the active ingredient due to the low partitioning of the same between the composition vehicle and the biological membrane to be permeated, which in the case of a nasal spray, would be the nasal mucosa.

[0049]

[0034] The aforementioned composition comprises lipophilic solvents and permeation enhancers that allow spraying in devices commonly used for intranasal administration, and has increased permeability, and consequently, bioavailability when compared to compositions that carry CBD only with solvents, usually PEG and ethanol. This improved bioavailability was obtained by the ideal mixture of transcutol / Labrasol permeation enhancers, below the amount usually used in pharmaceutical compositions, combined with solvents such as PEG, which, although not usually chosen as a vehicle for nasal solutions, has surfactant action and low viscosity, enabling the preparation under study with improved viscosity and permeation.

[0050]

[0035] With the aim of increasing absorption and reducing vehicle viscosity, a composition was devised containing two different concentrations of a permeation enhancer (transcutol, used up to 15% w / w in pharmaceutical compositions as a solubilizing agent) and a pharmaceutically acceptable vehicle for this type of composition, PEG - polyethylene glycol. Transcutol is normally used as a permeation enhancer in topical and transdermal compositions. Nasal permeation enhancers are generally found in the group of saponins, Laureth-9 (surfactant); fusidic acid derivatives, bile salts; oleic acid, caprylate, laurate (fatty acids); EDTA, salicylic acid (chelating agents); phospholipids, the use of transcutol being unusual. This ingredient is used as a desolubilizing agent, and there are no positive reports of its use as a permeation enhancer for intranasal administration [57,58].

[0051]

[0036] Sometimes, permeation enhancers can irritate the nasal mucosa and produce toxicity, which can be minimized by using suitable permeation enhancers at the appropriate concentration. Furthermore, the enhancer improves drug permeation in the vascular region, which often leads to serious peripheral side effects requiring adjustment of the amount and type of enhancer to be used. Considering the toxicity profile of transcutol (2-(2-ethoxyethoxy)ethanol), which is shown to be safe (up to 200g maximum daily intake) and minimally irritating to mucous membranes, this ingredient was chosen as the permeation enhancer for the target composition of this invention.

[0052]

[0037] With the target mixture of this invention, it is possible to obtain solutions that do not behave as nano-self-emulsifying systems or produce nanostructured systems, systems that have a different toxicity profile from that presented in simple solutions, an inventive principle explored here, allowing the use of cannabidiol via intranasal administration without the need for additional toxicity studies, which would be required in any other modified or nanostructured delivery system.

[0053]

[0038] Among the various molar mass range options available for polyethylene glycol (PEG), the one with the lowest viscosity, i.e., PEG 200, was chosen for the present invention. This choice was confirmed by plume formation tests using a placebo containing PEG 200 (Fig. 2) and PEG 400 (Fig. 1), in which only the use of PEG 200 as the main organic solvent promoted the formation of a consistent plume, as demonstrated in Figures 1 and 2, without significant weight variation with each application. Fig. 1 refers to the plume test applied to formulation FP2 and Fig. 2 refers to the plume test applied to formulation FP3. In both figures, the plume after activation can be observed on the left, and the representation of the physical parameters analyzed for regulatory purposes and affecting the product's efficiency is shown on the right. The product is the sum of the physical characteristics of the composition and the device.

[0054]

[0039] Furthermore, the surfactant action of PEG 200 aids the permeation of cannabidiol into the nasal mucosa, which, in association with Labrasol (a PEG derivative of medium-chain caprylic / capric fatty acid triglyceride), allows for an ideal balance between active ingredient partitioning between vehicle and nasal mucosa, viscosity, and adequate sprayability. A preservative, an antioxidant, and ethanol as an additional solubilizing agent were included in the composition. Labrasol had already been described as an excipient capable of increasing the bioavailability of cannabidiol in compositions for oral administration. In this sense, the use of the Labrasol-transcutol combination at the concentrations protected here was proposed as an ideal system for increasing nasal permeation of cannabidiol, aiming to increase its stability.

[0055] EXAMPLE OF IMPLEMENTATION

[0056]

[0040] The following table lists components and their concentration ranges present in the composition of the present invention. The concentrations of transcutol / Labrasol were varied in order to determine the ideal concentration of the mixture of components of the permeation-enhancing system.

[0057] Table 1: Preferred composition of nasal spray containing CBD. Raw material / Concentration range / component (%)

[0058] CBD 5 - 20

[0059] BHT 0.1 - 1

[0060] Transcutol 2 - 10

[0061] Labrasol 1 - 15

[0062] Tween 80 0.01 - 1

[0063] Cherry aroma 0.01 - 1

[0064] Vitamin E 0.01 - 1

[0065] Ethanol 1 - 15

[0066]

[0067] PEG 200 QSQ

[0041] For the evaluation of the pharmaceutical behavior of the compositions studied, 2 representative compositions were tested, composition a and composition b. The components and quantities are listed in tables 2 and 3 below:

[0068] Table 2: Preferred composition of nasal spray containing CBD - Formula a

[0069] Excipient / API Percentage (%) Amount (mg)

[0070] BD 5.0 ​​5000 BHT 0.2 200

[0071] Vitamin E 0.2 200 Transcutol 6.0 6000

[0072] Labrasol 10.0 10000 Tween (80) 0.04 40

[0073] Cherry aroma 0.01 10

[0074] Ethanol 8 8000 PEG 200 Qsp lOO Qsp lOO Total

[0075]

[0076] 100.0 100.0

[0077] Table 3: Preferred composition of nasal spray containing CBD formula b

[0078] Excipient / API Percentage (%) Amount (mg)

[0079]

[0080] CBD 10.0 5000 BHT 0.2 200

[0081] Vitamin E 0.2 200 Transcutol 6.0 6000

[0082] Labrasol 10.0 10000 Tween (80) 0.04 40 Cherry aroma 0.01 10

[0083] PEG 200 Qsp 100 Qsp 100 Total 100.0 100.0

[0084]

[0042] The process for preparing the composition includes the following steps:

[0085] A: Complete dissolution of BHT in ethanol and transcutol followed by the addition of CBD under agitation;

[0086] B: Complete dissolution of Tween 80 and benzalkonium chloride in ethanol under agitation; and

[0087] C: Mixing the solutions resulting from steps A and B, followed by the addition of PEG 200 to the mixture.

[0088] TEST AND RESULTS

[0089]

[0043] For the accelerated stability study, after the development and characterization of the compositions, the compositions aeb packaged in amber glass bottles with polyethylene screw caps were stored in a climate chamber under accelerated stability conditions (40 ± 2°C / 75 ± 5% RH). After 30, 60, and 90 days of stability, the compositions were evaluated for CBD content, pH, and density.

[0090]

[0044] Analysis of the CBD composition content at dosages of 10 mg / mL and 100 mg / mL after 30, 60 and 90 days of stability in a climate chamber (40° ± 2°C / 75 ± 5% RH) are presented in Tables 4 and 5 below.

[0091] Table 4 a - c: Analysis of the content of the compositions at 50 mg / mL and 100 mg / mL at time zero and after 30, 60 and 90 days of stability (40 ± 2°C / 75 ± 5% RH)

[0092] Composition a

[0093] Sample Time zero 30 days 60 days 90 days 1 99.23 95.78 98.43 97.46 2 99.20 98.47 103.26 99.67 3 99.89 97.40 97.49 95.87 4 100.45 101.71 101.20 98.32 5 97.46 101.80 101.70 94.61

[0094]

[0095] 6 100.91 103.37 101.64 97.77Average 99.52 99.76 100.62 97.28 SD 1.21 2.97 2.20 1.91 DPR

[0096]

[0097] 1.22 2.98 2.18 1.95 Density of CBD-containing compositions evaluated in the stability study - Composition a _

[0098] Analysis Time (days) Composition Density

[0099]

[0100] 0 1.07

[0101] 30 1.09

[0102] 60 1.10

[0103] 90 1.11

[0104] pH of the CBD-containing compositions evaluated in the stability study - Composition a

[0105] Analysis Time (days) pH

[0106]

[0107] 0 6.27

[0108] 30 6.31

[0109] 60 6.40

[0110] 90 6.38

[0111] Table 5 a - c: Analysis of the content of the compositions at 50 mg / mL and 100 mg / mL at time zero and after 30, 60 and 90 days of stability (40 ± 2°C / 75 ± 5% RH)

[0112] Composition b

[0113] Sample Time zero 30 days 60 days 90 days 1 99.98 96.18 95.13 93.10 2 99.20 97.57 96.16 91.36 3 99.02 98.40 97.04 92.47 4 100.01 99.21 98.29 95.52 5 98.76 98.43 97.01 93.31 6 100.02 97.32 95.33 90.97 Average 99.49 97.85 96.49 92.78 SD 0.78 0.83 0.96 1.49

[0114]

[0115] DPR 0.79 0.84 0.97 1.50 Density of CBD-containing compositions evaluated in the stability study - Composition b

[0116] Analysis Time (days) Composition Density

[0117]

[0118] 0 1.06

[0119] 30 1.02

[0120] 60 1.06

[0121] 90 1.02

[0122] pH of the CBD-containing compositions evaluated in the stability study - Composition b

[0123] Analysis Time (days) pH

[0124]

[0125] 0 6.21

[0126] 30 6.29

[0127] 60 6.43

[0128] 90 6.51

[0129]

[0045] Content assessment showed that the new compositions are stable and suitable for manufacturing cannabis-based medicines in the form of a nasal application solution.

[0130]

[0046] In order to evaluate the feasibility of applying the compositions of the present invention in devices normally used for nasal administration, two different types of devices were tested with the two compositions under test, with a placebo containing PEG 400, with a control in an ethanol / PEG 2001:1 mixture and with “composition a”, which uses corn oil and BHA in its composition. The devices used were the following: Aptar brand, Classic Line 140 uL model;

[0131] Aptar brand, Classic Line model, 50 uL; and RBR brand - standard model - 140 uL.

[0132]

[0047] It was found that in both prepared compositions, a and b (Fig. 3 and Fig. 4) the weight variation was much smaller than that observed for the use of PEG 400-placebo (Fig. 5) as well as for the composition containing vegetable oil (Fig. 6), indicating that PEG 200 was the ideal vehicle (excipient) to obtain a nasal spray composition without the use of special devices, with the weight variation observed for the control being statistically equal to that observed for compositions a and b.

[0133]

[0048] Thus, it was concluded that the aeb compositions are capable of forming a suitable plume without weight variation compared to commercial oil-based compositions, and that the results of these compositions, which contain PEG 200 and transcutol (high viscosity vehicle), permeation enhancers capable of improving the bioavailability of the compositions, are comparable to a low viscosity control sample, namely, the ethanol / PEG 200 1:1 mixture (Fig. 7). These results show the inventiveness of the composition and an unexpected effect in relation to the state of the art.

[0134]

[0049] In vivo model tests were performed by administering cannabidiol-containing compositions via nasal instillation to male Wistar rats weighing an average of 200g. Subsequently, blood samples were collected by cardiac puncture and plasma was isolated. The following permeation results were obtained for the compositions under study (FP2 and FP3), as shown in Tables 4 and 5.

[0135] Table 6: Plasma concentration and ASCo-o, 5h of cannabidiol for three compositions (control, FP25%, and FP35%) using the nasal instillation model in rats (N=3). Concentration ASCo-o, 5h Ratio Composition

[0136]

[0137] (pg / mL) (pgh / mL)

[0138] Control (50

[0139] 1.97 (±1.06) 0.49

[0140] mg / mL) 100%

[0141] 154%

[0142] Fa 10% 3.06 (± 1.83) 0.76

[0143] Fb 5% 3.13 (±1.36) 0.78 159%

[0144]

[0050] Thus, it was concluded that the aeb compositions present a higher concentration of cannabidiol in plasma compared to the control containing cannabidiol dissolved only in the PEG 200 / ethanol mixture, which indicates the superior bioavailability of said compositions, demonstrating the unexpected effect obtained with the new combination of permeation enhancers in the protected concentration range. The AUC (area under the curve) value from 0 to 30 minutes after administration, for the aeb compositions, was higher than the others, as seen in column 3 of table 6, confirming an increased bioavailability.

[0145]

[0051] The local irritability of these new compositions was evaluated using the RPMI 2650 cell line (ATCC® CCL-30) that simulates the nasal mucosa, evaluating pure CBD diluted in PEG 200 and composition a, with samples diluted in culture medium supplemented with fetal bovine serum (10%) and prepared at a concentration of 5 pg / mL. The sample representing the blank / vehicle and placebo was prepared using 20 pL of pure DMSO and 1980 pL of culture medium (representing 1% DMSO) and 20 pL of placebo and 1980 pL of culture medium, respectively.

[0146]

[0052] Immortalized human nasal epithelium cell line RPMI 2650 (ATCC® CCL-30), obtained from the Rio de Janeiro cell bank (BCRJ - 0412), was used for cytotoxicity evaluation. This cell line was cultured in EMEM culture medium supplemented with 10% fetal bovine serum and 1% streptomycin. The culture was maintained in a humidified incubator, 95% compressed air and 5% CO2 atmosphere. The culture was performed using 25 and 75 cm² flasks in supplemented EMEM medium, which was replaced every 48 hours. Once confluence was around 80%, cell passage was performed using a 0.25% trypsin-EDTA solution and divided into volumes of 5 or 8 ml of supplemented EMEM according to the respective flasks used. Cytotoxicity was assessed using the MTT (thiazolyl blue tetrazolium bromide) colorimetric method, using a 96-well plate and a cell density between 5-8 x 10 4cells / well. In each 96-well plate, the cells were distributed at a density of 8.0 x 10 4 Cells / well in a volume of 200 pL. The plates were kept in an incubator at 37°C containing 5% CO2 and 95% atmospheric air for 24h. In the next step, the culture medium of each cell was replaced with the test samples at a concentration of 5 ug / mL for both pure CBD and the test composition.

[0147]

[0053] After 24 h of incubation, the medium was replaced with 100 pL of MTT in PBS pH= 7.4 at a concentration of 0.5 mg / mL per well. The plates were covered with aluminum foil and incubated at 37°C containing 5% CO2 and 95% atmospheric air for 3 h. Then, the MTT solution was removed and 100 pL of pure, filtered DMSO was added to disrupt the cell membrane and solubilize the formazan crystals, generating a violet coloration of varying intensity depending on the degree of cell viability. Well readings were performed using a Microplate Absorbance Reader iMARK at 562 nm after vigorous shaking for 60 seconds.

[0148]

[0054] For the calculation of cell viability (Equation 1):

[0149] (Sample absorbance - blank absorbance) x 100 Viability (%) ~ ,■ ,< - > - — : — j - ; — f - r - z - > - — ~ 7 - ç

[0150]

[0151] to the negative two-absorbency white control)

[0152]

[0055] The variation in results was observed through statistical analysis applying ANOVA followed by Tukey's test for multiple comparisons using GraphPad Prism 5™ with a significance level of p < 0.05.

[0056] According to ISO 10993-5, samples can be considered non-cytotoxic with cell viability values ​​equal to or greater than 70% (ISO, 2009). Thus, we can observe in the cytotoxicity assay by the MTT colorimetric method, CBD and composition a, at a concentration of 5 pg / mL (Figure 8), are considered non-cytotoxic in relation to the negative control, confirming the tolerability of the permeation enhancer mixture used in the target compositions of the invention described herein.

[0153]

[0057] The aeb compositions were tested in terms of their ability to undergo dilution with water or 0.9% w / v saline solution in order to increase their biocompatibility and patient acceptance for chronic use. The aeb compositions were then modified by removing PEG 200 and diluting with 20% 0.9% w / v saline solution in order to assess whether the new compositions obtained would remain as true solutions with improved stability and bioavailability. The same composition of the permeation enhancer mixture was maintained in the ced compositions described in Tables 7 and 8.

[0154] Table 7: Preferred composition of CBD-containing nasal spray - Formula c

[0155] Percentage Amount Excipient / API

[0156] (%) (mg)

[0157]

[0158] CBD 5.0 ​​5000

[0159] Vitamin E 0.2 200

[0160] BHT 0.2 200

[0161] Transcutol 6.0 6000

[0162] Labrasol 10.0 10000

[0163] Tween (80) 0.04 40

[0164] Cherry flavor 0.01 10 Saline solution 0.9%

[0165] 25.0 25000

[0166] p / v

[0167] Ethanol q.s.p. 100 q.s.p. 100

[0168] Total 100.0 100.0

[0169] Table 8: Preferred composition of nasal spray containing CBD formula d

[0170] Percentage Amount Excipient / API

[0171] (%) (mg)

[0172] CBD 10.0 5000

[0173] Vitamin E 0.2 200

[0174] BHT 0.2 200

[0175] Transcutol 6.0 6000

[0176] Labrasol 10.0 10000

[0177] Tween (80) 0.04 40

[0178] Aroma of ce

[0179]

[0180] reja 0,01 10

[0181] 0.9% saline solution

[0182] 20.0 20000

[0183] p / v

[0184] Ethanol q.s.p. 100 q.s.p. 100

[0185] Total 100.0 100.0

[0186]

[0058] After dilution, the ced compositions remain as true solutions, which was confirmed in the determination of the droplet size distribution using a dynamic light scattering -DLS instrument, with no formation of nanostructured systems observed as a result of this dilution (Figure 9).

[0187]

[0059] The new spray obtained had its plume-forming capacity evaluated, and in both compositions the weight variation proved to be low, less than 10 pg per application, with a more uniform plume than that obtained with the undiluted compositions, as observed in Fig. 10 and Fig. 11. Fig. 10 and Fig. 11 demonstrate the plume formed using composition c and composition d, respectively. In both figures, on the left, the plume after one activation can be observed, and on the right, the representation of the physical parameters analyzed for regulatory purposes and that affect the product's efficiency. The product is the sum of the physical characteristics of the composition and those of the device.

[0188]

[0060] Table 9 summarizes the physical parameters demonstrating the importance of the physicochemical characteristics of the formulation combined with the type of actuator / valve / device, which directly affect the actuation result (as shown in Figures 1, 2, 10 and 11). Table 9: Response of the combination of the formulation with the nasal spray device containing CBD

[0189] Angle Angle Diameter Diameter Height Height Formulation of 1 o of 2 o from 1 o of 2 o from 1 o of 2 o cone cone cone cone cone cone FP2 20° N / A 9 cm N / A 27 cm N / A FP3 10° 40° 0.5 cm 9 cm 14 cm 14 cm Fc 40° N / A 9.8 cm N / A 21 cm N / A Fd 20° N / A 6.5 cm N / A 18 cm N / A

[0190]

[0191]

[0061] The stability of the CBD compositions was evaluated at 40°C / 75% RH, with a content decay of less than 10% relative to the initial value for both. The permeation study was conducted for a composition containing 5% CBD, following the same experimental procedure previously used to evaluate the undiluted compositions, with the observed concentration value of 3.72 pg / mL (±1.12) very close to the highest value obtained for the undiluted compositions, 3.13 (±1.36), confirming the improved bioavailability promoted by the new protected permeation enhancement system in this invention.

[0192]

[0062] It should be understood that the present description does not limit the application to the details described herein and that the innovation is capable of other embodiments and of being practiced or performed in a variety of ways, within the scope of the claims. Although specific terms have been used, such terms should be interpreted in a generic and descriptive sense, and not for the purpose of limitation.

Claims

MODIFIED CLAIMS Received by the International Secretariat on April 24, 2026 (24.04.2026) 1. NASAL SPRAY COMPOSITION, characterized by being in the form of a solution and comprising cannabidiol, a combination of: - polyethylene glycol 200, ethanol or saline solution of 0.1 to 1.5% w / v; - between 2 and 20%, by weight, of cannabidiol; - between 0.1 and 1%, by weight, of Butyl Hydroxytoluene; - between 0.1 and 1%, by weight, of Vitamin E; - between 1 and 10%, by weight, of anionic surfactant Polyoxyethylene (20) Sorbitan Monooleate; - between 0.01 and 1%, by weight, of Benzalkonium HCl; - between 1 and 20%, by weight, of Ethanol; - between 0.01 and 1%, by weight, of aroma; - between 0.1% and 15% by weight of caprylocaproyl macrogol-8 glycerides; - between 0.1% and 20% by weight of ethoxydiglycol; and - between 0.1% and 25% by weight of 0.9% w / v saline solution.

2. “USE OF A COMPOSITION”, as defined in claim 1, characterized by being for preparing a medicament that assists in the treatment of anxiety / stress, depression, chronic pain, inflammation, sleep disorders, epilepsy and neuroprotection. MODIFIED SHEET (ARTICLE 19)

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