WO2023130142A2 - Compositions and methods using cannabinoid compounds for treating urinary tract disorders - Google Patents

Abstract

Methods of treating disorders of the urinary tract system using cannabinoid compounds are described herein. The cannabinoid compounds can be selected from the group of cannabigerol ("CBG"), cannabigerovarinic acid ("CBGVA"), cannabidiol ("CBD"), (+)-cannabidiol ("(+)-CBD"), cannabigerovarin ("CBGV"), cannabicyclol ("CBL"), cannabichromene ("CBC"), cannabigerol butyl ("CBG-C4"), tetrahydrocannabivarin ("THCV"), and combinations thereof. Compositions and articles including the cannabinoid compounds are further disclosed.

Description

COMPOSITIONS AND METHODS USING CANNABINOID COMPOUNDS FOR TREATING URINARY TRACT DISORDERS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the priority benefit of U.S. Provisional Patent App. Serial No. 63/226,361, filed January 3, 2022, which is hereby incorporated by reference herein.

TECHNICAL FIELD

[0002] The present disclosure generally relates to the use of cannabinoid compounds and related products to alleviate symptoms and treat various urinary tract disorders.

BACKGROUND

[0003] The human urinary tract system includes the kidneys, ureters, bladder, and urethra. Medical disorders in the urinary tract system can cause pain, discomfort, and interfere with normal urination. Such medical disorders can include urinary tract infections (“UTIs”), interstitial cystitis, and bladder incontinence. Several types of UTIs are classified with infections in the lower urinary tract system specifically referred to as a bladder infection or cystitis, infections in the upper urinary tract as a kidney infection or pyelonephritis, and infections in the urethra as urethritis. UTIs occur when an infectious agent enters the urinary tract, typically through the urethra, and progresses to cause an infection. The most common infectious agent is Escherichia coli (“E. coli”). Related disorders can present similar symptoms as UTIs without an underlying infection including interstitial cystitis or painful bladder. Additionally, bladder incontinence can occur due to a UTI, interstitial cystitis, or can occur due to other conditions such as stress, constipation, and the consumption of various foods and drinks.

DETAILED DESCRIPTION

[0004] Disorders of the urinary tract system can be painful, debilitating, and embarrassing. For example, inflammation of the bladder, also called cystitis, typically presents as pelvic pressure, lower abdomen discomfort, frequent and painful urination, and as blood in urine. Similarly, bladder incontinence can cause a restriction in activities and severe embarrassment. The severity and lifestyle impacts of such urinary tract disorders often requires medical treatment. If the disorders are chronic, ongoing treatments can be required.

[0005] Although cystitis is most frequently caused by a UTI where an infectious agent spreads into the bladder typically through the urethra, cystitis can also occur without an infection. When cystitis occurs without an underlying infection, the disorder is called interstitial cystitis or painful bladder syndrome.

[0006] Treatment of cystitis varies depending upon the underlying cause. For example, cystitis caused by a UTI can be treated with antibiotics if the underlying infectious agent is susceptible. For example, a bladder infection caused by E. coli is commonly treated with antibiotics such as trimethoprim/sulfamethoxazole, fosfomycin, nitrofurantoin, cephalexin, or ceftriaxone.

[0007] As can be appreciated however, treatment of cystitis can be more difficult in certain circumstances including when the infectious agent is resistant to antibiotic treatment and particularly when the cystitis is interstitial cystitis. Interstitial cystitis does not have a clearly known underlying cause making both prevention and treatment of interstitial cystitis more difficult.

[0008] Similarly, bladder incontinence can be caused by a variety of underlying causes including UTIs, interstitial cystitis, stress, constipation, pregnancy, age, menopause, enlarged prostate or prostate cancer, obstructions, and neurological disorders. The diversity of causes of bladder incontinence makes treatment difficult particularly for transient causes or in situations where it is caused by a difficult to treat UTI or interstitial cystitis.

[0009] Accordingly, there is an unmet need for products to alleviate symptoms and treat multiple bladder conditions particularly when the underlying cause is undiagnosed or during the duration of time while the underlying cause is being treated. Compositions, articles, products, methods and treatments for both cystitis and bladder incontinence are described herein. In certain embodiments, the cannabinoid compounds described herein can alleviate symptoms of cystitis and bladder incontinence by one or more of reducing the concentration of prostaglandin E2 (“PGE2”) and by modulation of certain G-protein coupled receptors (“GPCR” or “GPC receptors”) in the bladder. In certain embodiments, the compositions, articles, products, methods, and treatments can both inhibit production of PGE2 and can modulate certain GPC receptors.

[0010] Reduction of PGE2 and modulation of the GPCR system in the bladder can alleviate the symptoms of multiple bladder disorders because these systems are both known mechanisms that cause the symptoms of bladder disorders. For example, multiple studies have suggested that the body’s response to an UTI is activation of the cyclooxygenase-2 (“COX-2”)/PGE2 signaling pathway. It is theorized that inhibition of this signaling pathway would alleviate the symptoms of a UTI and provide relief from the UTI. Additional studies have found that PGE2 levels also rise in individuals with Hunner’s lesions, a form of interstitial cystitis. Cannabinoid compounds discovered to act as PGE2 signaling inhibitors include cannabigerol (“CBG”) and cannabigerovarinic acid (“CBGVA”).

[0011] Additional studies provide evidence that the compounds used in the regulation of the COX- 2/PGE2 signaling pathway can also provide relief for other bladder disorders including bladder incontinence. For example, studies have found that muscarinic acetylcholine receptor Ml (“CHRMi”) antagonists can reduce bladder contractility. CHRMi antagonists include CBG, cannabidiol (“CBD”), (+)-cannabidiol (“(+)-CBD”), cannabigerovarin (“CBGV”), cannabicyclol (“CBL”), and cannabichromene (“CBC”). As can be appreciated, reducing bladder contractility can decrease the severity of bladder incontinence.

[0012] In addition to the CHR i receptor, studies have also indicated that endocannabinoid receptor 2 (“CB2R”), but not endocannabinoid receptor 1 (“CBiR”), is linked to cystitis including interstitial cystitis. Specifically, in various studies, interstitial cystitis was induced in mice through intravesical instillation of acrolein or through lipopolysaccharide and then evaluated against various CBiR and CB2R modulators. The studies found that CB2R agonists, but not CBiR agonists, decreased the severity of interstitial cystitis. Cannabinoid compounds which can act as CB2R agonists include CBC, (+)-CBD, cannabigerol-C4 (“CBG-C4”), and tetrahydrocannabivarin (“THCV”).

[0013] Yet other studies have suggested the potential for cannabinoid compounds to alleviate and treat bladder disorders. For example, one study compared treatment of a UTI with THC and a cannabis extract and found that cannabis extract performed better than THC alone. Although a cause of action was not determined, the study suggested that cannabinoid compounds of relatively high abundance in cannabis extract can be effective in alleviating and treating bladder disorders.

[0014] Cannabinoid compounds discovered to alleviate symptoms and treat bladder disorders include CBG, CBGVA, CBD, (+)-CBD, CBGV, CBL, CBC, CBG-C4, and THCV. However, it has also been found that certain cannabinoid compounds do not have a known mechanism to reduce bladder disorders are not expected to be effective in reducing the severity of any bladder disorders. Examples of such cannabinoid compounds include cannabinol (“CBN”), cannabidiolic acid (“CBDA”), cannabigerolic acid (“CBGA”), cannabidivarin (“CBDV”), cannabidiethanol (“CBD-C2”), cannabidiorcol (“CBD-C1”), cannabidivarinic acid (“CBDVA”), cannabigerolic acid butyl (“CBGA-C4”), cannabichromenic acid (“CBCA”), and cannabicyclolic acid (“CBLA”).

[0015] Prior to the present discovery, it was not appreciated which specific cannabinoid compounds could be used to treat bladder disorders. For example, prior attempts at addressing bladder disorders did not focus on specific cannabinoid compounds, or focused only on THC, and did not test individual cannabinoid compounds as described in the present disclosure. By selecting only cannabinoid compounds that have mechanisms of action, the methods, compositions, articles and products described herein can provide improved treatment of bladder conditions with targeted specificity.

[0016] The present discovery was facilitated by the Applicant’s unique methods of producing hereto rare cannabinoid compounds in appreciable quantities including through chemical synthesis reactions and growth in yeast cultures. Prior to the Applicant’s research, the lack of viable production of individual cannabinoid compounds obviated the ability to treat gastric acid diseases with specific cannabinoid compounds. Additional details about the production of producing rare cannabinoid compounds are described in PCT Patent Application Nos. WO 2020/069142 Al, WO 2020/069214 A2, WO 2021/05597 Al; and WO 2020/236789 Al, each of which is incorporated herein by reference.

[0017] Prior to the Applicant’s process of isolating specific and unique cannabinoid compounds from non-horti cultural sources, cannabinoid compounds were extracted and isolated only from naturally grown marijuana plants which drastically limited the volume of the rarer cannabinoid compounds available for research or use. Thus, these non-horticulturally-derived cannabinoid compounds offer benefits in regard to the treatment of gastric acid buildup not previously contemplated. As used herein, non-horticulturally derived cannabinoid compounds refers to cannabinoid compounds not grown in plants (e.g., not through horticulture or agriculture).

[0018] Additionally, isolated cannabinoid compounds extracted from marijuana plants can also suffer from purity issues as certain unavoidable containments (such as other natural marijuana plant compounds, irremovable amounts of other cannabinoid compounds, etc.) can remain present in isolated cannabinoid compounds extracted from marijuana plants. Such unavoidable containments can impact the quality of the data or even alter the apparent functioning of the cannabinoid compounds. Compositions and methods of alleviating bladder disorders that use horticulturally derived cannabinoid compounds may not exhibit the same effects as compositions and methods using purer cannabinoid compounds such as the cannabinoid compounds contemplated herein. As can be appreciated however, horticulturally derived cannabinoid compounds can be used in certain embodiments of the disclosure if the horticulturally extracted cannabinoid compounds are sufficiently pure and/or if any containments are sufficiently well understood.

[0019] By inhibiting the production of PGE2 or modulating the CHRMi or CB2R receptors, the cannabinoid compounds described herein can reduce or eliminate the symptoms caused by various urinary tract disorders, such as bladder disorders. For example, the severity of bladder discomfort can be lessened or normal urination frequency restored. In certain embodiments, the compositions or products containing the herein described cannabinoid compounds can be administered or used on a predetermined schedule to treat chronic bladder disorders. Additionally, or alternatively, such compositions or products can be administered or used on an as-needed schedule when bladder issues occur. In certain embodiments, the cannabinoid compounds described herein can be used in conjunction with other treatments, such as antibiotic treatment, to provide relief of symptoms while the underlying cause of the bladder disorder is treated.

[0020] Generally, treatment of a bladder disorder can be accomplished by treatment with a therapeutically effective amount of one or more of CBG, CBGVA, CBD, (+)-CBD, CBGV, CBL, CBC, CBG-C4, and THCV. In certain embodiments, each of CBG, CBGVA, CBD, (+)-CBD, CBGV, CBL, CBC, CBG-C4, and THCV can be included in a composition, article or product while in other embodiments, various combinations and subsets of CBG, CBGVA, CBD, (+)-CBD, CBGV, CBL, CBC, CBG-C4, and THCV can be included in such compositions, articles and products.

[0021] For example, in certain embodiments, only one cannabinoid compound, such as CBG, can be included. As can be appreciated however, any combination of CBG, CBGVA, CBD, CBC, and THCV can be effective including the use of just a single cannabinoid compound selected from the foregoing cannabinoid compounds.

[0022] A therapeutically effective amount of the one or more cannabinoid compounds can vary depending on factors such as the desired effect of treatment, the severity of the bladder disorder, the duration of treatment, or the method of delivering the cannabinoid compounds to the subject. For example, severe pain caused by interstitial cystitis may require a greater amount of the one or more cannabinoid compounds than bladder incontinence.

[0023] In certain embodiments, a therapeutically effective amount for internal use (e.g., an oral or injectable composition) can be about 100 mg of the cannabinoid compounds or less; in certain embodiments, about 75 mg of the cannabinoid compounds or less; in certain embodiments, about 50 mg of the cannabinoid compounds or less; in certain embodiments, about 20 mg of the cannabinoid compounds or less; in certain embodiments, about 10 mg of the cannabinoid compounds or less; in certain embodiments, about 5 mg of the cannabinoid compounds or less; in certain embodiments, about 1 mg of the cannabinoid compounds or less; in certain embodiments, about 500 pg of the cannabinoid compounds or less; in certain embodiments, about 100 pg of the cannabinoid compounds or less; and in certain embodiments, about 500 pg of the cannabinoid compounds or less.

[0024] As can be appreciated, the relative concentration of the cannabinoid compounds can vary in different compositions and products. For example, a beverage containing the cannabinoid compounds can have a smaller concentration of the cannabinoid compounds than a pill or capsule. In certain embodiments however, the total amount of the cannabinoid compounds can be the same between such two compositions and articles. In other embodiments, both the concentration and amount of cannabinoid compounds can vary between different compositions and articles. [0025] In certain embodiments, the relative amounts of each of CBG, CBGVA, CBD, (+)-CBD, CBGV, CBL, CBC, CBG-C4, and THCV can vary in the compositions and articles described herein. For example, each individual cannabinoid compound (CBG, CBGVA, CBD, (+)-CBD, CBGV, CBL, CBC, CBG-C4, and THCV) can vary from each other cannabinoid compound by about 1,000: 1 to about 1 : 1,000. As can be appreciated, the amount and ratios of each of the cannabinoid compounds can be selected based on factors such as the method of delivery and individual factors such as the body weight of person consuming the cannabinoid compounds.

[0026] In certain embodiments, the compositions, articles, and methods described herein can be substantially or entirely free of cannabinoid compounds other than of C CBG, CBGVA, CBD, (+)- CBD, CBGV, CBL, CBC, CBG-C4, and THCV. For example, the compositions, articles, products and methods can be substantially or entirely free of tetrahydrocannabinol (“THC”). As used herein, substantially free can mean less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, less than about 0.5%, less than about 0.1%, or less than about 0.01%. In certain embodiments, the cannabinoid compounds can be produced using non-horticulturally- derived methods such as through chemical synthesis (e.g., organic synthesis reactions) or through modification of yeast and/or bacterial cells to produce the cannabinoid compounds in high purity. However, in certain embodiments, cannabinoid compounds can also be a natural product, e.g., an extract of a cannabis plant if sufficiently pure. In certain embodiments, substantially pure means that the isolated cannabinoid compound, when added, includes about 3% or less of contaminants, about 2% or less of contaminants, about 1% or less of contaminants, about 0.5% or less of contaminants, about 0.1% or less of contaminants, or about 0.01% or less of contaminants.

[0027] In certain embodiments, the compositions, articles, products and methods described herein can be utilized on a predetermined schedule (e.g., nightly, twice daily, etc.) or can be utilized on an as-needed basis. In certain embodiments, the predetermined schedule can be based on the halflife of the cannabinoid compounds as well as the release dynamics of the cannabinoid compounds. As can be appreciated, it can be useful in certain embodiments, to release the cannabinoid compounds described herein using a delayed release mechanism, such as a delayed release pill, to regulate the bioavailable amounts of the cannabinoid compounds. [0028] In certain embodiments, the cannabinoid compounds described herein can be included in a composition, article or product to partially or fully alleviate diseases or symptoms caused by a bladder disorder. The composition, article or product can be consumed by, or be applied onto, a person to alleviate the urinary tract disorder Generally, the exact nature of the composition, article or product can vary widely.

[0029] For example, the cannabinoid compounds can be included in pills or capsules that can be taken quickly and efficiently on a predetermined basis or as-needed basis (daily, with meals, etc.). Use of an oral pill or capsule can be useful in the treatment of a bladder disorder. As can be appreciated, pills and capsules can contain a number on inactive ingredients as known in the art such as dicalcium phosphate dehydrate, microcrystalline cellulose, stearic acid, silicon dioxide, croscarmellose sodium, magnesium stearate, and pharmaceutical glaze. Other known pills and capsules are also contemplated herein. As an additional example, a compressed chewable tablet can include a water-disintegrable, compressible carbohydrate (such as mannitol, sorbitol, maltitol, dextrose, sucrose, xylitol, lactose and mixtures thereof), a binder (such as cellulose, cellulosic derivatives, polyvinyl pyrrolidone, starch, modified starch and mixtures thereof), the cannabinoid compounds and, optionally, a lubricant (such as magnesium stearate, stearic acid, talc, and waxes), sweetening, coloring and flavoring agents, a surfactant, a preservative, and other ingredients. All of the ingredients, including the one or more cannabinoid compounds, are dry blended and compressed into a tablet.

[0030] In certain embodiments, the cannabinoid compounds can alternatively be administered to individuals via food products and other comestibles. By way of illustration and not as a limitation, the selected cannabinoid compounds can be incorporated into a beverage, a “smoothie” (fruit, vegetable, nut oil, or yogurt based), a frozen desert (e.g., ice cream or sorbet), a food bar, a nutrition bar, a dressing, a snack, into a flour- or flour-alternative-based product, a rice-based product, pastes, gels, powders, gums, etc. Incorporation into food products can facilitate consumption of the cannabinoid compounds and increase palatability.

[0031] As can be appreciated, the exact nature of the food article can influence the bioavailability of the cannabinoid compounds. For example, a cannabinoid compound included in a large food article may take more time to become bioavailable than the same amount of cannabinoid compound in a single pill or capsule. Generally, the remainder of the composition, article or product can constitute any suitable non-bioactive component such as filler, food, or water.

[0032] In certain embodiments, the compositions, articles or products including the cannabinoid compounds described herein can include indicia and/or packaging to convey to end users the amount of the cannabinoid compounds contained therein. For example, a small nutrient bar may be individually labeled and packaged to express to the end user that only a single bar should be consumed.

[0033] As will be appreciated, a wide variety of different compositions, articles and products can be prepared which include the one or more cannabinoid compounds of the present disclosure including compositions, articles and products not listed here. All such compositions, articles and products are contemplated herein as they are within the ordinary skill of artisans based on the guidance provided in the present disclosure.

[0034] Generally, all of the compositions, articles and products described herein can be manufactured and produced as known in the art. For example, in certain embodiments, the cannabinoid compounds can be dissolved in a suitable solvent such as an alcohol or oil and then added to the composition, article or product.

Examples

[0035] Abbreviations used herein include the following:

DMEM Dulbecco’s modified Eagle’s medium

EGF Epidermal growth factor

ELISA Enzymelinked immunosorbent assay

FCS Fetal calf serum

GAM Goat anti-mouse

IL Interleukin

MIC Minimum inhibitory concentration

MW Molecular weight

NHEK Normal human epidermal keratinocytes

NHDF Normal human dermal fibroblasts OD Optical density

PBS Phosphate buffered saline

PE Pituitary extract

PGE2 Prostaglandin E2

PM A Phorbol myristate acetate

ROS Reactive oxygen species

RT Room temperature

SFM Serum free medium

TGF Transforming growth factor

UV Ultraviolet

Xem Emission wavelength

Xex Excitation wavelength

[0036] Example 1: Inhibition of PGE2

[0037] Example 1 demonstrates the effects of Compounds A, B, C, D, E and L-Ascorbic on inhibiting the release of PGE2. As noted in table below for test compounds, Compound A is cannabigerol (CBG); Compound B is cannabidiol (CBD); Compound C is cannabigerovarinic acid (CBGVA); Compound D is cannabidiolic acid (CBDA); and Compound E is cannabigerolic acid (CBGA).

[0038] PGE2 release was measured on normal human epidermal keratinocytes (NHEK) by PMA- stimulated NHEK using specific ELISA kits, and reactive oxygen species (ROS) production by UV-irradiated NHEK. The quantification of ROS was performed using a fluorescent probe (2,7- dichlorodihydrofluorescein diacetate).

[0039] The effects of the test compounds on normal human dermal fibroblasts (NHDF) were measured by assessing: procollagen I synthesis in “aged” fibroblasts obtained by replicative senescence according to the Hayflick model using a specific ELISA kit, IL-8 release by IL- la- stimulated NHDF using a specific ELISA kit.

[0040] Materials and Methods Biological models

Human dermal fibroblasts:

Test Compounds:

[0041] PGE2 release by PMA-stimulated NHEK

Culture and treatments

[0042] Keratinocytes were seeded in 96-well plates and cultured for 24 hours in culture medium. The culture medium was then replaced by assay medium 1 containing or not (control) the test compounds or the reference compound (indomethacin tested at 1 pM for PGE2 release) and the cells were pre-incubated for 24 hours. After pre-incubation, the medium was removed and replaced by assay medium 1 containing or not (stimulated control) the compounds or the reference compound and containing the inducer (PMA tested at 0.5 pg/ml). The cells were then incubated for 24 hours. In parallel, a non-stimulated control condition was performed.

[0043] All experimental conditions were performed in n=3. At the end of incubation, the culture supernatants were collected for PGE₂ quantifications. [0044] Enzyme-Linked Immunosorbent Assay (ELISA)

[0045] PGE2 released in the culture supernatants were measured using specific ELISA kits according to the supplier’s instructions.

[0046] ROS production in UV-irradiated NHEK

Culture and treatments

[0047] Keratinocytes were seeded in 96-well plates and cultured for 24 hours in culture medium and then in assay medium 2 for a further 24 hours. The culture medium was then removed and replaced by assay medium 2 containing or not (irradiated control) the test compounds or the reference compound (Vitamin E tested at 100 pM) and the cells were pre-incubated for 24 hours. After pre-incubation, the medium was removed andreplaced by assay medium 2 containing the fluorescent probe, 2, 7-di chlorodihydrofluorescein diacetate (2,7-DCDHF-DA in assay medium) and the cells were incubated for 30 minutes at 37°C. The cells were then washed and recovered with a PBS solution. The cells were then irradiated with UVB (+ UVA) - 100 mJ/cm² (+0.7 J/cm²). The lamp used was a SOL500 Sun Simulator equipped with an H2 filter (Dr. Hbnle, AG). After irradiation, the medium was replaced by a PBS solution containing or not (irradiated control) the test compounds or the reference, and the cells were incubated for 30 minutes. A nonirradiated control and a condition with no probe (background noise) were performed in parallel. All experimental conditions were performed in n=3.

Quantification of ROS production

[0048] The emitted fluorescence intensity (+ex = 485 nm, Xem = 538 nm) was measured using a Synergy Hl (BioTek) microplate reader. The fluorescence intensity of the metabolized probe was proportional to the formation of ROS. The production of ROS was therefore expressed as relative fluorescence intensity. [0049] Procollagen I synthesis in “aged” fibroblasts

Culture and treatment

[0050] The “aged” (P18-F) fibroblasts were seeded in 96-well plates and cultured for 24 hours in culture medium. The culture medium was then replaced by assay medium containing or not (control) the test compounds or the reference compound (TGF-P tested at 10 ng/ml) and the cells were incubated for 72 hours.

[0051] All experimental conditions were performed in n=3. At the end of incubation, the culture supernatants were collected for procollagen I quantification.

[0052] Enzyme-Linked Immunosorbent Assay (ELISA)

[0053] Procollagen I released in the culture supernatants was measured using a specific ELISA kit according to the supplier’s instructions.

IL-8 release by IL- la-stimulated NHDF

Culture and treatment

[0054] Fibroblasts were seeded in a 96-well plate and cultured for 24 hours in culture medium. The culture medium was then replaced by assay medium containing or not (stimulated control) the test compounds or the reference compound (IKK Inhibitor X tested at 10 pM) and the cells were pre-incubated for 24 hours. After pre-incubation, the culture medium was removed and replaced by assay medium containing or not (stimulated control) the test compounds or the reference compound and containing the inducer (IL-la tested at 0.1 ng/ml). The cells were then incubated for 24 hours. In parallel, a non-stimulated control was performed.

[0055] All experimental conditions were performed in n=3. At the end of incubation, the culture supernatants were collected for IL-8 quantification. [0056] Enzyme-Linked Immunosorbent Assay (ELISA)

[0057] IL-8 released in the culture supernatants was measured using a specific ELISA kit according to the supplier’s instructions.

Data management

[0058] Raw data were analyzed using Microsoft Excel® software and GraphPad PRISM® software.

[0059] The inter-group comparisons were performed by an unpaired Student’s t-test. The statistical analysis can be interpreted if n>5, however for n<5 the statistical values are for information only.

[0060] Results

[0061] Effect on PGE2 release by PMA-stimulated keratinocytes

[0062] The effects of compounds A, B, C, D, and E and of L-ascorbic acid on PGE2 release are summarized in Table 1.

Table 1: Effect of Compounds A, B, C, D, E and L-Ascorbic acid on PGE2 release by PMA-stimulated keratinocytes

[0063] In the non-stimulated control condition, a low basal level of PGE2 was secreted by NHEK (62 pg/ml). The treatment of NHEK with PMA at 0.5 pg/ml for 24 hours resulted in a significant stimulation of PGE2 release (96 pg/ml) and this effect was completely inhibited by the reference compound indomethacin, tested at 1 pM (>169% of relative inhibition). These results were expected and validated the assay.

[0064] Compounds A (CBG), C (CBGVA) and L-Ascorbic acid strongly and significantly inhibited PGE2 release by PMA-stimulated keratinocytes (about 160%, 160% and 145% of relative inhibition, respectively). Regarding compound A (CBG), the inhibitory effect was significant at all tested concentrations and equivalent when tested at 1 and 3 pM. Compound C (CBGVA) had a significant effect only when tested at the highest concentration. Finally, L- Ascorbic acid induced overall the same effect at all tested concentrations.

[0065] On the other hand, Compounds B (CBD) and E (CBGA) overstimulated PGE2 release by PMA-stimulated keratinocytes reaching respectively 185% and 182% of the control when tested at the highest concentration. Finally, Compound D (CBDA) did not modulate PGE2 release by PMA-stimulated keratinocytes.

[0066] Example 2: GPCR Testing

[0067] To evaluate CHRMi antagonistic activity and CB2R agonist activity of the cannabinoid compounds, a GPCR reactivity assay was performed. The evaluated cannabinoid compounds were: (+)-CBD, CBD, CBDV, CBD-C2, CBD-C1, CBG, CBG-C4, CBGV, CBC, CBL, CBN, THCV, CBDA, CBDVA, CBGA, CBGA-C4, CBGVA, CBCA, and CBLA. Sauvagine was used as the control for CHRMi while CP55940 (a synthetic THC analogue) was used as the control for CB2R.

[0068] To perform the GPCR reactivity assay, a commercial GPCR assay, PathHunter® P- Arrestin from Eurofins DiscoverX Products (Fremont, CA), was used. In the PathHunter® P- Arrestin GPCR assay, an inactive peptide fragment is fused to the targeted GPCR receptor and a complementary peptide fragment is fused to P-arrestin. At activation of the GPCR receptor and recruitment of P-arrestin, complementation of the peptide fragments occurs and restores P- galactosidase activity. The amount of P-galactosidase activity is then measurable using chemiluminescent reagents.

[0069] A single assay was run for each of CHRMi and CB2R. For CHRMi, in each run, PathHunter® cell lines were removed from a freezer stock and seeded at a volume of 20 pL into white walled, 384-well microplates and incubated at 37 °C. Each cell was pre-incubated with the cannabinoid compound followed by an agonist challenge at the EC80 concentration. Cells were then diluted to generate a 5x sample in assay buffer. 5 pL of the 5x sample was then added to cells and incubated at 37 °C for 30 minutes. Finally, 5 pL of 6x EC80 agonist in assay buffer were added to the cells and incubated at 37 °C for 90 minutes or 180 minutes. The assay signal was generated through addition of 12.5 pL or 15 pL (50% V/V) of a detection reagent cocktail followed by a one hour incubation time at room temperature. Microplates were read following signal generation with a PerkinElmer Envision™ (PerkinElmer Inc., Waltham, MA) for chemiluminescent signal detection. Reactivity was analyzed using the CBIS data analysis suite (Chemlnnovation, CA) where RLU refers to the raw measured values. The percent inhibition was calculated using the formula: % Inhibition = 100% * (1 - mean RLU of test sample - mean RLU of control) / (mean RLU of EC80 control - mean RLU of vehicle control).

[0070] For CB2R, PathHunter® cell lines were removed from a freezer stock and seeded at a volume of 20 pL into white walled, 384-well microplates and incubated at 37 °C. Each cell was incubated with a sample to induce a response and then diluted to generate a 5x sample in assay buffer. 5 pL of the 5x sample was then added to cells and incubated at 37 °C for 90 or 180 minutes. The final assay concentration was 1%. The assay signal was generated through addition of 12.5 pL or 15 pL (50% V/V) of a detection reagent cocktail followed by a one hour incubation time at room temperature. Microplates were read following signal generation with a PerkinElmer Envision™ (PerkinElmer Inc., Waltham, MA) for chemiluminescent signal detection. Reactivity was analyzed using the CBIS data analysis suite (Chemlnnovation, CA) where RLU refers to the raw measured values. The percent activity was then calculated using the formula: % Activity = 100% * (mean RLU of test sample - mean RLU of control) / (mean max control ligand - mean RLU of vehicle control).

[0071] The results of the GCPR reactivity screen are depicted in Table 2. The percent inhibition is depicted for CHRMi antagonism and the percent activation for CB2R agonism are depicted.

TABLE 2

TABLE 2 (Continued)

[0072] As depicted in Table 2, CBD, (+)-CBD, CBGV, CBL, and CBC inhibited CHRMi and would be expected to reduce bladder contractions while CBC, (+)-CBD, CBG-C4, and THCV activated CB2R and would be expected to reduce the symptoms of cystitis.

[0073] The dimensions and values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value.

[0074] It should be understood that every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written herein. Every minimum numerical limitation given throughout this specification will include every higher numerical limitation, as if such higher numerical limitations were expressly written herein. Every numerical range given throughout this specification will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein.

[0075] Every document cited herein, including any cross-referenced or related patent or application, is hereby incorporated herein by reference in its entirety unless expressly excluded or otherwise limited. The citation of any document is not an admission that it is prior art with respect to any invention disclosed or claimed herein or that it alone, or in any combination with any other reference or references, teaches, suggests, or discloses any such invention. Further, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in the document shall govern.

[0076] The foregoing description of embodiments and examples has been presented for purposes of description. It is not intended to be exhaustive or limiting to the forms described. Numerous modifications are possible in light of the above teachings. Some of those modifications have been discussed and others will be understood by those skilled in the art. The embodiments were chosen and described for illustration of various embodiments. The scope is, of course, not limited to the examples or embodiments set forth herein, but can be employed in any number of applications and equivalent articles by those of ordinary skill in the art. Rather it is hereby intended the scope be defined by the claims appended hereto.

[0077] It should be understood that certain aspects, features, structures, or characteristics of the various embodiments can be interchanged in whole or in part. Reference to certain embodiments mean that a particular aspect, feature, structure, or characteristic described in connection with certain embodiments can be included in at least one embodiment and may be interchanged with certain other embodiments. The appearances of the phrase “in certain embodiments” in various places in specification are not necessarily all referring to the same embodiment, nor are certain embodiments necessarily mutually exclusive of other certain embodiments. It should also be understood that the steps of the methods set forth herein are not necessarily required to be performed in the orders described, and the order of the steps of such methods should be understood to be merely exemplary. Likewise, additional steps can be included in such methods, and certain steps may be omitted or combined, in methods consistent with certain embodiments.

Claims

WHAT IS CLAIMED IS:

1. A composition for treating disorders of the urinary tract system, the composition comprising one or more cannabinoid compounds.

2. The composition of claim 1, wherein the composition therapeutically treats one or more of a urinary tract infection (“UTT’), interstitial cystitis, and bladder incontinence.

3. The composition of claim 1, wherein the one or more cannabinoid compounds are selected from the group of cannabigerol (“CBG”), cannabigerovarinic acid (“CBGVA”), cannabidiol (“CBD”), (+)-cannabidiol (“(+)-CBD”), cannabigerovarin (“CBGV”), cannabicyclol (“CBL”), cannabichromene (“CBC”), cannabigerol butyl (“CBG-C4”), and tetrahydrocannabivarin (“THCV”), and combinations thereof.

4. The composition of any preceding claim, wherein the one or more cannabinoid compounds exhibit one or more of: inhibition of the cyclooxygenase-2 (“COX-2”)/PGE2 signaling pathway; antagonistic activity against the muscarinic acetylcholine receptor Ml (“CHRMi”); and agonist activity against endocannabinoid receptor 2 (“CB2R”).

5. The composition of claim 1 comprises about 100 mg or less of the one or more cannabinoid compounds.

6. The composition of claim l is a liquid.

7. The composition of claim l is a food or beverage.

8. The composition of claim 1 is a pill or capsule.

9. The composition according to claim 8 is a delayed-release pill or capsule.

10. The composition of claim 1, wherein the one or more cannabinoid compounds are non- horticulturally derived cannabinoid compounds.

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11. An article comprising the composition of any of the preceding claims.

12. A method of treating a disorder of the urinary tract system, the method comprising administering a therapeutically effective amount of one or more cannabinoid compounds, wherein the one or more cannabinoid compounds are selected from the group of cannabigerol (“CBG”), cannabigerovarinic acid (“CBGVA”), cannabidiol (“CBD”), (+)-cannabidiol (“(+)-CBD”), cannabigerovarin (“CBGV”), cannabicyclol (“CBL”), cannabichromene (“CBC”), cannabigerol butyl (“CBG-C4”), tetrahydrocannabivarin (“THCV”), and combinations thereof.

13. The method of claim 12, wherein the disorder comprises one or more of a urinary tract infection (“UTT’), interstitial cystitis, and bladder incontinence.

14. The method of claims 12 to 13, further comprising the step of administering an antibiotic medicine.

15. The method of claims 12 to 13, wherein the one or more cannabinoid compounds are administered on a predetermined schedule.

16. The method of any of claims 12 to 13, wherein the one or more cannabinoid compounds are administered on an as-needed schedule.

17. The method of claims 12 to 13, wherein the one or more cannabinoid compounds are non- horticulturally derived cannabinoid compounds.