IL300307A - Compositions of cannabinoids and methods of using same - Google Patents

Description

GYNI-P-002-IL COMPOSITIONS OF CANNABINOIDS AND USE THEREOF FIELD OF THE INVENTION The present invention relates to compositions of cannabinoids, and use thereof, such as for inhibiting secretion of a pro-inflammatory cytokine, reducing cyclooxygenase expression or nitric oxide production, and for treatment or prevention of a disease, e.g., an inflammatory disease.

BACKGROUND Endometriosis, present in 10% of reproductive-age women, is a disease of the female reproductive system in which normal endometrial mucosa (glands and stroma) abnormally grow outside the uterus. Common symptoms include pelvic pain, heavy periods, pain with bowel movements, and infertility. Painful sexual intercourse, also known as dyspareunia, occurs in nearly half of women with endometriosis, while in 70%, pain occurs during menstruation. Painful periods or menstrual cramps are also known as dysmenorrhea. The main symptom of dysmenorrhea is pain concentrated in the lower abdomen or pelvis co-occurring with nausea and vomiting, diarrhea or constipation, headache, dizziness, disorientation, hypersensitivity to sound, light, smell and touch, fainting, and fatigue. The use of certain types of birth control pills can prevent the symptoms of dysmenorrhea because they stop ovulation from occurring. Other medications that may help also include nonsteroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen, hormonal birth control, and the intrauterine device (IUD) with progestogen. Surgery may be useful if certain underlying problems are present.

Although the pathogenesis of endometriosis is not clearly understood, dysregulation of the immune system in the endometriotic milieus has been considered to play a pivotal role. Macrophage recruitment and nerve fiber infiltration are the two major characteristics of this aberrant immune environment. The recruitment of macrophages within the endometriotic lesion has been demonstrated to facilitate the development and maintenance of endometriosis. The infiltration of nerve fibers and their abnormal GYNI-P-002-IL distribution has been shown to be involved in the generation of endometriosis-associated pain and inflammatory response.

As of today, endometriosis is still an unmet clinical need and the search for novel treatment options is challenging. The current treatment options for endometriosis are surgical removal of endometriotic lesions, drug therapy or a combination of both. The drug treatments for endometriosis are based on anti-hormonal drugs that suppress ovulation and the release of reproductive hormones, such as estrogen, the hormone that contributes to the growth of endometriotic lesions. Currently available drugs for endometriosis are not equally effective in patients with different forms of the disease, and due to their adverse effects, most therapies are recommended for a limited length of time. This is a major problem as these hormonal medications only temporarily suppress endometriosis symptoms and do not eradicate the disease. Therefore, the recurrence rate of pelvic pain is high, with estimation of 40–50% within five years after cessation of therapy. Painkillers, such as morphine sulphate tablets or other opioid painkillers, and anti-inflammatory drugs (e.g., NSAIDs such as diclofenac and pentoxifylline) are commonly used in conjunction with other therapy to provide pain control and to reduce inflammation.

Among potential drug candidates, endocannabinoids and modulators of the endocannabinoid system are emerging as potential pharmacological targets in women’s disease, as they affect specific mechanisms that are critical to disease establishment and maintenance, especially the level of pain.

PCT Publication No. WO2021240510A1 describes the use of a cannabinoid and its respective acid precursor, such as cannabidiol (CBD) and cannabidiolic acid (CBDA), or tetrahydrocannabinol (THC) and tetrahydrocannabinol acid (THCA), in treating a subject afflicted with an IL-6-related disease, a COX-related disease, or both, such as endometriosis, and endometriosis-related symptoms. Nonetheless, the use of specific flavonoids for treating these diseases, and particularly, the desired ratio between a cannabinoid and a flavonoid, for such treatments, has not been addressed so far.

GYNI-P-002-IL SUMMARY The present invention, in some embodiments, is based, at least in part, on the surprising finding that i n - v i t r o treatment of stimulated monocyte/macrophage cell line with a composition comprising a cannabinoid, such as cannabidiol (CBD) or cannabigerol (CBG), and a flavonoid, such as apigenin (e.g., in a molar ratio of 2:1), synergistically inhibits the inflammatory response, as compared to its respective mono-treatment (Fig. 3). Unexpectedly, the combination of a cannabinoid and a flavonoid, as disclosed herein, inhibited to a higher extent the pro-inflammatory response as compared to dual treatment with a cannabinoid and its respective acid precursor (e.g., CBD and CBDA, Fig. 3). The invention is further based on the surprising finding that a composition comprising CBG and the cannabinoid precursor CBDA (e.g., in a molar ratio of 1:1), was more effective in decreasing IL-6 secretion and NO production, as compared to a composition comprising CBG and the CBD (Fig. 3 and Fig. 6, respectively). Among the dual treatments, the combinations of: (i) CBD and apigenin, (ii) CBG and apigenin, and (iii) CBG and CBDA, were also found to a have a the most profound anti-inflammatory effect in the context of IL-1β secretion (Fig. 4), TNF-α secretion (Fig. 5), and nitic oxide (NO) production (Fig. 6). The present invention, in some embodiments, is further based, at least in part, on the finding that addition of a psychotropic cannabinoid, e.g., THC, THCA, and THCV, substantially diminished NO secretion (Fig. 6) and decreased cyclooxygenase (COX) expression, with higher specificity to COX-2 (Fig. 2). These results indicate an anti-inflammatory therapeutic potential for these compositions in pro-inflammatory diseases evolving macrophage activation, pro-inflammatory cytokines secretion and COX dysregulation, such as endometriosis, and endometriosis-related symptoms.

According to one aspect there is provided a pharmaceutical composition comprising: a cannabinoid selected from cannabidiol (CBD) or cannabigerol (CBG); and apigenin, wherein the CBD or CBG and the apigenin are present in the pharmaceutical composition in a weight per weight (w/w) or mole per mole (m:m) ratio ranging from 1:1 to 10:1.

GYNI-P-002-IL According to another aspect there is provided a pharmaceutical composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) a cannabinoid selected from CBD and CBG; and (ii) apigenin.

According to another aspect, there is provided a pharmaceutical composition comprising a first active ingredient and a carrier, wherein the first active ingredient consists essentially of CBDA and CBG.

In some embodiments, the cannabinoid is CBG.

In some embodiments, the CBD or CBG and the apigenin are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 10:1.

In some embodiments, the CBD or CBG and apigenin are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 4:1.

In some embodiments, the pharmaceutical composition further comprises cannabidiolic acid (CBDA).

In some embodiments, the CBD or CBG and the CBDA are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 2:1 to 1:2.

In some embodiments, the pharmaceutical composition being any one of: (i) the cannabinoid is CBD, and the pharmaceutical composition further comprises CBG; and (ii) the cannabinoid is CBG, and the pharmaceutical composition further comprises CBD.

In some embodiments, the CBD and the CBG are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 2:1 to 1:2.

In some embodiments, the pharmaceutical composition further comprises an additional compound selected from the group consisting of: tetrahydrocannabinol (THC), tetrahydrocannabinol acid (THCA), tetrahydrocannabivarin (THCV), and any combination thereof.

In some embodiments, the CBD or CBG and the additional compound are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 4:1.

In some embodiments, the THC and the THCA are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 2:1 to 2.5:1.

GYNI-P-002-IL In some embodiments, the CBDA and the CBG are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 1:10 to 10:1.

In some embodiments, the pharmaceutical composition further comprises a second active ingredient, wherein the second active ingredient consists essentially of apigenin.

In some embodiments, the CBDA and the CBG are present in the composition in a w/w or m:m ratio ranging from 1:2 to 2:1.

In some embodiments, the CBDA or CBG and the apigenin are present in the pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 10:1.

In some embodiments, the pharmaceutical composition is for use in the treatment or prevention of an interleukin-6 (IL-6)-related disease, an IL-1β- related disease, a tumor necrosis factor α (TNF-α)-related disease, a cyclooxygenase (COX)-related disease, a nitric oxide (NO)-related disease, or any combination thereof.

In some embodiments, the disease is selected from the group consisting of: an autoimmune disease, an inflammatory disease, endometriosis, dysmenorrhea, and dyspareunia.

In some embodiments, COX is COX2.

Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

GYNI-P-002-IL In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by reference to the study of the following detailed description.

BRIEF DESCRIPTION OF THE FIGURES Fig. 1 includes a vertical bar graph showing cyclooxygenase 1 (COX1) and COXexpression in activated RAW 246.7 cells pre-treated with cannabinoids. Expression of COX1 and COX2 was examined by Western blot and was normalized to β actin. The graph exhibits COX1 and COX2 expression as compared to a positive control sample (100%); activated cells pre-treated with vehicle alone. Non-activated cells without any pre-treatment served as a negative control sample. THC (tetrahydrocannabinol); THCA (tetrahydrocannabinolic acid); CBD (cannabidiol); THCV (tetrahydrocannabivarin); CBG (cannabigerol); CBC (cannabichromene); (β-CP) β caryophyllene; apigenin.

Fig. 2 includes a vertical bar graph showing COX1 and COX2 expression in activated RAW 246.7 cells pre-treated with different ratios of THC:THCA. Expression of COX1 and COX2 was examined by Western blot and was normalized to β actin. The graph exhibits COX1 and COX2 expression as compared to a positive control sample (100%); activated cells pre-treated with only vehicle. Non-activated cells without any pre-treatment served as a negative control sample.

Fig. 3 includes a vertical bar graph showing IL-6 secretion from activated RAW 246.7 cells pre-treated with different combinations of cannabinoids and/or flavonoids as described in the X axis of the graph. The number given in brackets indicates the concentration in µM. Graph depicts the % of IL-6 secretion in each sample from positive LPS activated control cells. Red lines depict 100% and 50% as reference.

Fig. 4 includes a vertical bar graph showing interleukin 1β (IL-1β) secretion from activated RAW 246.7 cells pre-treated with different combinations of cannabinoids and/or flavonoids as described in the X axis of the graph. The number given in brackets indicates the concentration in µM. Graph depicts the % of IL-1β secretion in each sample from positive LPS activated control cells. Red lines depict 100% and 50% as reference.

GYNI-P-002-IL Fig. 5 includes a vertical bar graph showing tumor necrosis factor α (TNF-α) secretion from activated RAW 246.7 cells pre-treated with different combinations of cannabinoids and/or flavonoids as described in the X axis of the graph. The number given in brackets indicates the concentration in µM. Graph depicts the % of TNF-α secretion in each sample from positive LPS activated control cells. Red lines depict 100% and 50% as reference.

Fig. 6 includes a vertical bar graph showing nitric oxide (NO) secretion from activated RAW 246.7 cells pre-treated with different combinations of cannabinoids and/or flavonoids as described in the X axis of the graph. The number given in brackets indicates the concentration in µM. Graph depicts the % of NO secretion in each sample from positive LPS activated control cells. Red lines depict 100% and 50% as reference.

DETAILED DESCRIPTION Compositions According to some embodiments, there is provided a composition comprising a cannabinoid or a precursor acid thereof, and a flavonoid, wherein the cannabinoid or a precursor thereof, and the apigenin are present in the composition in a weight per weight (w/w) or mole per mole (m:m) ratio ranging from 1:1 to 100:1. In some embodiments, the flavonoid comprises apigenin. In some embodiments, the cannabinoid is a non-psychotropic cannabinoid.

As used herein, the terms “non-psychotropic” and “non-psychoactive” are used interchangeably and are well known in the art. A non-psychotropic cannabinoid encompasses a cannabinoid that does not affect a subject’s behavior, mood, thoughts, perception, or any combination thereof, when consumed by the subject and reaches his central nervous system (CNS). In some embodiments, a non-psychotropic cannabinoid is selected from: cannabidiol (CBD), cannabigerol (CBG), cannabinol (CBN), cannabichromene (CBC), cannabidivarin (CBDV), a precursor acid thereof, or any combination thereof. In some embodiments, a non-psychotropic cannabinoid is CBD, CBG, or both.

In some embodiments, a precursor comprises an acid precursor of a cannabinoid.

GYNI-P-002-IL In some embodiments, there is provided a composition comprising a cannabinoid and apigenin, being present in the composition in a weight per weight (w/w) or mole per mole (m:m) ratio ranging from 1:1 to 100:1.

In some embodiments, there is provided a composition comprising a precursor of a cannabinoid and apigenin, being present in the composition in a weight per weight (w/w) or mole per mole (m:m) ratio ranging from 1:1 to 100:1, wherein the cannabinoid precursor is an acid.

In some embodiments, the composition comprises apigenin and a cannabinoid selected from: CBD or CBG.

In some embodiments, the composition comprises apigenin and a cannabinoid precursor selected from: CBDA or CBGA.

In some embodiments, the weight per weight (w/w) or molar (molar/molar) ratio of CBD or CBG to apigenin within the composition as described herein is in the range of 1:10 to 10:1. In some embodiments, the w/w or molar/molar ratio of CBD or CBG to apigenin within the composition is in the range of 1:10 to 10:1, 1:9 to 9:1, 1:8 to 8:1, 1:7 to 7:1, 1:6 to 6:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, or 1:2 to 2:1. Each possibility represents a separate embodiment of the invention.

In some embodiments, the w/w or m:m ratio of CBD or CBG to apigenin within the composition as described herein is in the range of 1:1 to 10:1. In some embodiments, the w/w or m:m ratio of CBD or CBG to apigenin within the composition is in the range of 1:to 10:1, 1:1 to 9:1, 1:1 to 9:1, 1:1 to 8:1, 1:1 to 7:1, 1:1 to 6:1, 1:1 to 5:1, 1:1 to 4:1, or 1:to 3:1. Each possibility represents a separate embodiment of the invention.

In some embodiments, the w/w or m:m ratio of CBD or CBG to apigenin within the composition as described herein is in the range of 1:1 to 4:1. In some embodiments, the w/w or m:m ratio of CBD or CBG to apigenin within the composition is about 2:1.

In some embodiments, the composition comprises CBD and apigenin. In some embodiments, CBD and apigenin are present in the composition in a w/w or m:m ratio ranging from 1:1 to 4:1. In some embodiments, CBD and apigenin are present in the composition in a w/w or m:m ratio of about 2:1.

GYNI-P-002-IL In some embodiments, the composition comprises CBG and apigenin. In some embodiments, CBG and apigenin are present in the composition in a w/w or m:m ratio ranging from 1:1 to 4:1. In some embodiments, CBG and apigenin are present in the composition in a w/w or m:m ratio of about 2:1.

In some embodiments, the composition further comprises cannabidiolic acid (CBDA). In some embodiments, the composition comprises CBD, apigenin, and CBDA. In some embodiments, the composition comprises CBG, apigenin, and CBDA.

In some embodiments, the w/w or m:m ratio of CBD or CBG to CBDA within the composition as described herein is in the range of 10:1 to 1:10. In some embodiments, the w/w or m:m ratio of CBD or CBG to CBDA within the composition is in the range of 1:to 10:1, 1:9 to 9:1, 1:8 to 8:1, 1:7 to 7:1, 1:6 to 6:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, or 1:to 2:1. Each possibility represents a separate embodiment of the invention. In some embodiments, the w/w or m:m ratio of CBD or CBG to CBDA within the composition is in the range of 2:1 to 1:2. In some embodiments, the w/w or m:m ratio of CBD or CBG to CBDA within the composition is in the range of about 1:1.

In some embodiments, the cannabinoid is CBD, and the composition further comprises CBG. In some embodiments, the cannabinoid is CBG, and the composition further comprises CBD. In some embodiments, the composition comprises CBD, CBG, and a flavonoid. In some embodiments, the composition comprises CBD, CBG, and apigenin. In some embodiments, the composition comprises CBD, CBG, CBDA, and apigenin.

In some embodiments, the w/w or m:m ratio of CBD to CBG within the composition as described herein is in the range of 10:1 to 1:10. In some embodiments, the w/w or m:m ratio of CBD to CBG within the composition is in the range of 10:1 to 1:10, 1:to 9:1, 1:8 to 8:1, 1:7 to 7:1, 1:6 to 6:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, or 1:2 to 2:1. Each possibility represents a separate embodiment of the invention. In some embodiments, the w/w or m:m ratio of CBD to CBG within the composition is in the range of 2:1 to 1:2. In some embodiments, the w/w or m:m ratio of CBD to CBG within the composition is about 1:1.

In some embodiments, the composition disclosed herein further comprises a psychotropic cannabinoid.

GYNI-P-002-IL As used herein, the terms “psychotropic” and “psychoactive” are interchangeable and encompass a cannabinoid that affects a subject’s behavior, mood, thoughts, perception, or any combination thereof, when consumed by the subject and consumed by the subject and reaches his CNS. In some embodiments, a psychotropic cannabinoid is selected from: tetrahydrocannabinol (THC, or ∆9THC), tetrahydrocannabinol acid (THCA), tetrahydrocannabivarin (THCV), ∆8THC, or any combination thereof. In some embodiments, a psychotropic cannabinoid is selected from: THC, THCA, THCV, or any combination thereof.

In some embodiments, the composition further comprises an additional compound selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the composition comprises CBD, apigenin, and an additional compound selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the composition comprises CBG, apigenin, and an additional compound selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the composition comprises CBD, CBG, apigenin, and an additional compound selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the composition comprises CBD, CBG, apigenin, CBDA and an additional compound selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the additional compound disclosed herein comprises one compound selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the additional compound disclosed herein comprises two compounds selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the additional compound comprises THC, THCA and THCV.

In some embodiments, the w/w or m:m ratio of CBD or CBG to the additional compound as disclosed herein is in the range of 1:1 to 10:1. In some embodiments, the w/w or m:m ratio of CBD or CBG to at least one of: THC, THCA, and THCV, within the composition as described herein is in the range of 1:1 to 10:1. In some embodiments, the w/w or m:m ratio of CBD or CBG to at least one of: THC, THCA, and THCV, within the composition is in the range of 1:1 to 10:1, 1:1 to 9:1, 1:1 to 8:1, 1:1 to 7:1, 1:1 to 6:1, 1:1 to 5:1, 1:1 to 4:1, or 1:1 to 3:1. Each possibility represents a separate embodiment of the invention. In some embodiments, the w/w or m:m ratio of CBD or CBG to at least one of: THC, THCA, and THCV, within the composition is in the range of 1:1 to 4:1. In some GYNI-P-002-IL embodiments, the w/w or m:m ratio of CBD or CBG to at least one of: THC, THCA, and THCV, within the composition is in the range of 2:1.

In some embodiments, the w/w or m:m ratio of apigenin to at least one of: THC, THCA, and THCV, within the composition as described herein is in the range of 1:10 to 10:1. In some embodiments, the w/w or m:m ratio of apigenin to at least one of: THC, THCA, and THCV, within the composition is in the range of 1:10 to 10:1, 1:9 to 9:1, 1:8 to 8:1, 1:7 to 7:1, 1:6 to 6:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, or 1:2 to 2:1. Each possibility represents a separate embodiment of the invention. In some embodiments, the w/w or m:m ratio of apigenin to at least one of: THC, THCA, and THCV, within the composition is in the range of 1:2 to 2:1. In some embodiments, the w/w or m:m ratio of apigenin to at least one of: THC, THCA, and THCV, within the composition is about 1:1.

In some embodiments, the w/w or m:m ratio of THC to THCA within the composition disclosed herein is in the range of 1:1 to 15:1, 1:1 to 14:1, 1:1 to 13:1, 1:1 to 12:1, 1:1 to 11:1, 1:1 to 10:1, 1:1 to 9:1, 1:1 to 8:1, 1:1 to 7:1, 1:1 to 6:1, 1:1 to 5:1, 1:1 to 4:1, 1:1 to 3:1, or 1:1 to 2:1. Each possibility represents a separate embodiment of the invention. In some embodiments, the w/w or m:m ratio of THC to THCA within the composition is in the range of 1:1 to 4:1, 1:1 to 3:1, or 1:1 to 2:1. In some embodiments, the w/w or m:m ratio of THC to THCA within the composition is in the range of 2:1 to 2.5:1. In some embodiments, the w/w or m:m ratio of THC to THCA within the composition disclosed herein is 7:3.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) a cannabinoid selected from CBD, CBG, or both, and (ii) apigenin.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) CBD, and (ii) apigenin.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) CBG, and (ii) apigenin.

GYNI-P-002-IL The term "consists essentially of" denotes that a given compound or substance constitutes the vast majority of the active ingredient's portion or fraction of the composition.

In some embodiments, consists essentially of means that the combination of: (i) CBD, CBG, or both, and (ii) apigenin constitutes at least 95%, at least 98%, at least 99%, or at least 99.9% by weight, of the active ingredient(s) of the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) a cannabinoid selected from CBD, CBG, or both, (ii) apigenin; and (iii) CBDA.

In some embodiments, consists essentially of means that the combination of (i) a cannabinoid selected from CBD, CBG, or both, (ii) apigenin; and (iii) CBDA, constitutes at least 95%, at least 98%, at least 99%, or at least 99.9% by weight, of the active ingredient(s) of the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) a cannabinoid selected from: CBD, CBG, or both (ii) apigenin; and (iii) an additional compound selected from: THC, THCA, THCV, or any combination thereof.

In some embodiments, consists essentially of means that the combination of (i) a cannabinoid selected from CBD, CBG, or both, (ii) apigenin; and (iii) an additional compound selected from: THC, THCA, THCV, or any combination thereof, constitutes at least 95%, at least 98%, at least 99%, or at least 99.9% by weight, of the active ingredient(s) of the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) a cannabinoid selected from CBD, CBG, or both, (ii) apigenin, (iii) CBDA; and (iv) an additional compound selected from: THC, THCA, THCV, or any combination thereof.

GYNI-P-002-IL In some embodiments, consists essentially of means that the combination of (i) a cannabinoid selected from CBD, CBG, or both, (ii) apigenin, (iii) CBDA; and (iv) an additional compound selected from: THC, THCA, THCV, or any combination thereof, constitutes at least 95%, at least 98%, at least 99%, or at least 99.9% by weight, of the active ingredient(s) of the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the additional compound disclosed herein comprises one compound selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the additional compound comprises two compounds selected from: THC, THCA, THCV, or any combination thereof. In some embodiments, the additional compound comprises THC, THCA and THCV.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of: (i) a cannabinoid selected from CBD, CBG, or both; and (ii) CBDA. In some embodiments, the active ingredient consists essentially of CBD and CBDA. In some embodiments, the active ingredient consists essentially of CBG and CBDA.

In some embodiments, consists essentially of means that the combination of: (i) a cannabinoid selected from CBD, CBG, or both; and (ii) CBDA, constitutes at least 95%, at least 98%, at least 99%, or at least 99.9% by weight, of the active ingredient(s) of the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of a first active ingredient, being at least one of: CBD, CBG, CBDA, or any combination thereof, and a second active ingredient being apigenin.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of a first active ingredient being CBD and CBDA, and a second active ingredient being apigenin.

In some embodiments, there is provided a composition comprising an active ingredient and a carrier, wherein the active ingredient consists essentially of a first active ingredient being CBG and CBDA, and a second active ingredient being apigenin.

GYNI-P-002-IL In some embodiments, the composition disclosed herein further comprises a terpen. In some embodiments, the terpen is selected from: β caryophyllene (BCP), β myrcene, or any combination thereof.

In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

In some embodiments, the composition is formulated for dermal administration. In some embodiments, the composition is formulated for vaginal administration. In some embodiments, the composition is formulated for systemic administration. In some embodiments, the composition is formulated for rectal administration. In some embodiments, the composition is formulated for abdominal administration. In some embodiments, the composition is formulated for subcutaneous administration. In some embodiments, the composition is formulated for intra-peritoneal administration. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition is formulated for administration to a subject.

The term "pharmaceutically acceptable" means suitable for administration to a subject, e.g., a human. For example, the term "pharmaceutically acceptable" can mean approved by a regulatory agency of the Federal or a state government or listed in the U. S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic compound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates or GYNI-P-002-IL phosphates. Antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; and agents for the adjustment of tonicity such as sodium chloride or dextrose are also envisioned.

The route of administration of the composition will depend on the disease or condition to be treated. Suitable routes of administration include, but are not limited to, parenteral injections, e.g., intradermal, intravenous, intramuscular, intralesional, subcutaneous, intrathecal, and any other mode of injection as known in the art. Although the bioavailability of the active ingredients to be administered by other routes can be lower than when administered via parenteral injection, by using appropriate compositions it is envisaged that it will be possible to administer the compositions of the invention via transdermal, oral, rectal, vaginal, topical, nasal, inhalation and ocular modes of treatment. In addition, it may be desirable to introduce the pharmaceutical composition of the invention by any suitable route, including intraventricular and intrathecal injection; intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer.

In some embodiments, the composition is a pharmaceutical composition.

In some embodiments, the composition is for use in the treatment of an inflammatory disease. In some embodiments, the inflammatory disease is characterized by or comprises a high level of a pro-inflammatory mediator. In some embodiments, a pro-inflammatory mediator comprises a pro-inflammatory cytokine. In some embodiments, a pro-inflammatory mediator is selected from: interleukin 6 (IL-6), IL-1β, tumor necrosis factor α (TNF-α), nitric oxide (NO), cyclooxygenase (COX), or any combination thereof. In some embodiments, expression and/or secretion of the pro-inflammatory mediator is increased. In some embodiments, the composition is for use in the treatment of an IL-6-related disease, an IL-1β-related disease, a TNF-α-related disease, a COX-related disease, a NO- related disease, or any combination thereof, in a subject in need thereof.

In some embodiments, COX is COX1, COX2, or both. In some embodiments, COX comprises COX2. In some embodiments, COX is COX2.

GYNI-P-002-IL Methods of use In some embodiments, there is provided a method for reducing or inhibiting a pro-inflammatory mediator expression and/or secretion by an immune cell, comprising contacting the immune cell with an effective amount of the composition of the invention, thereby reducing or inhibiting the pro-inflammatory mediator expression and/or secretion by the immune cell. In some embodiments, the pro-inflammatory mediator comprises a pro-inflammatory cytokine. In some embodiments, the pro-inflammatory cytokine comprises a cytokine selected from: IL-6, IL-β, TNF-α, or any combination thereof. In some embodiments, the pro-inflammatory mediator is selected from: IL-6, IL-β, TNF-α, nitric oxide (NO), COX, or any combination thereof. In some embodiments, COX comprises COX1, COX2, or both.

As used herein, the term “pro-inflammatory mediator” encompasses any specialized substance, such as a signaling molecule, an enzyme, a vasoactive amine or peptide, an eicosanoid, a proinflammatory cytokine, and an acute-phase protein, that mediates the inflammatory process.

As used herein, the term "immune cell" refers to any cell of the host defense system within an organism which protects against disease, pathogens, other pathological agents or abnormalities, or divergence from homeostasis.

In some embodiments, the immune cell is obtained or derived from a subject.

As used herein, the term “subject” refers to any subject, including a mammalian subject, for whom therapy is desired, for example, a human.

In some embodiments, the subject is a human subject.

In some embodiments, the immune cell is selected from: a monocyte a macrophage, a lymphocyte, or a mast cell.

In some embodiments, the lymphocyte is a B lymphocyte, a T lymphocyte, or both.

In some embodiments, contacting is contacting in vivo, ex-vivo or in vitro. In some embodiments, ex-vivo or in vitro comprises or is in a test tube or in a plate.

GYNI-P-002-IL Methods for determining expression and/or secretion levels of a pro-inflammatory mediator, e.g., IL-6, IL-β, TNF-α, NO, and COX, are common and would be apparent to one of ordinary skill in the art. Non-limiting examples for methods of determining expression and/or secretion levels of a pro-inflammatory mediator include, but are not limited to, immunoassays, such as enzyme-linked immunosorbent assay (ELISA), western blot, dot-blot, MS-MS, or others.

In some embodiments, there is provided a method for treating a subject afflicted with an IL-6-related disease, an IL-1β-related disease, a TNF-α-related disease, a COX-related disease, a NO- related disease, or any combination thereof, comprising administering to the subject a therapeutically effective amount of the composition of the invention, thereby treating an IL-6-related disease, an IL-1β-related disease, a TNF-α-related disease, a COX-related disease, a NO- related disease, or any combination thereof, in the subject.

In some embodiments, the method further comprises a step of determining the expression and/or secretion level of IL-6, IL-1β, TNF-α, COX, NO, or any combination thereof, wherein expression and/or secretion level any one of IL-6, IL-1β, TNF-α, COX, NO, or any combination thereof, above a predetermined threshold, indicates the subject is suitable for treatment with the composition of the invention.

In some embodiments, expression and/or secretion level any one of IL-6, IL-1β, TNF-α, COX, NO, or any combination thereof, below or equal to a predetermined threshold, indicates the subject is not suitable for treatment with the composition of the invention.

In some embodiments, the determining step is performed before the administering step. In some embodiments, the method further comprises a determining step being performed after the administering step. In some embodiments, the method comprises at least a first determining step being performed before the administering step and at least a second determining step being performed after the administering step.

In some embodiments, there is provided a method for selecting a subject being suitable for treatment with the composition of the invention, comprising the steps of: (a) determining the expression and/or secretion level of IL-6, IL-1β, TNF-α, COX, NO, or any combination thereof, wherein expression and/or secretion level of any one of the IL-6, IL-1β, TNF-α, COX, NO, or any combination thereof, above a predetermined threshold GYNI-P-002-IL indicates the subject is suitable for treatment with the composition of the invention, and (b) administering to a subject determined to be suitable for treatment according to step (a) a therapeutically effective amount of the composition of the invention.

In some embodiments, the determining step is performed in the subject or in a sample derived or obtained from the subject. In some embodiments, the sample comprises any bodily fluid, cell, tissue, biopsy, organ, or a combination thereof, derived or obtained from the subject. In some embodiments, the determining step is performed in vivo, ex vivo, or in vitro.

Methods and means for in vitro and/or ex vivo assays, as described herein, are common and would be apparent to one of ordinary skill in the art.

In some embodiments, ex vivo or in vitro comprises or is in a test tube or in a plate.

In some embodiments, an IL-6-related disease, an IL-1β-related disease, a TNF-α-related disease, a COX-related disease, a NO- related disease, or any combination thereof, comprises or is an immune disease, including, but not limited to inflammation.

In some embodiments, an IL-6-related disease, an IL-1β-related disease, a TNF-α-related disease, a COX-related disease, a NO- related disease, or any combination thereof, comprises or is an immune disease, including, but not limited to inflammation.

In some embodiments, an IL-6-related disease, an IL-1β-related disease, a TNF-α-related disease, a COX-related disease, a NO- related disease, or any combination thereof, comprises or is a disease involving angiogenesis pathology, increased growth factor expression and/or secretion, endothelial dysfunction, endothelial cell migration and/or proliferation, or any combination thereof.

As used herein, the term “IL-6-related disease" refers to any pathology or a disease wherein IL-6 expression, secretion, or both, initiates, propagates, involved, promotes, enhances, triggers, or any combination thereof, a pathological condition, or a disease or a symptom thereof. In some embodiments, IL-6-related disease is any disease or condition involving IL-6 expression, secretion, or both, in the pathogenesis, pathophysiology, or both, of the disease or condition.

GYNI-P-002-IL As used herein, the term “IL-1β-related disease" refers to any pathology or a disease wherein IL-1β expression, secretion, or both, initiates, propagates, involved, promotes, enhances, triggers, or any combination thereof, a pathological condition, or a disease or a symptom thereof. In some embodiments, IL-1β-related disease is any disease or condition involving IL-1β expression, secretion, or both, in the pathogenesis, pathophysiology, or both, of the disease or condition.

As used herein, the term “TNF-α-related disease" refers to any pathology or a disease wherein TNF-α expression, secretion, or both, initiates, propagates, involved, promotes, enhances, triggers, or any combination thereof, a pathological condition, or a disease or a symptom thereof. In some embodiments, TNF-α-related disease is any disease or condition involving TNF-α expression, secretion, or both, in the pathogenesis, pathophysiology, or both, of the disease or condition.

As used herein, the term “COX-related disease" refers to any pathology or a disease wherein COX expression, secretion, or both, initiates, propagates, involved, promotes, enhances, triggers, or any combination thereof, a pathological condition, or a disease or a symptom thereof. In some embodiments, COX-related disease is any disease or condition involving COX expression, secretion, or both, in the pathogenesis, pathophysiology, or both, of the disease or condition. In some embodiments, COX-related disease is COX1-, COX2-, or both- related disease. In some embodiments, COX-related disease comprises COX2-related disease. In some embodiments, COX-related disease is COX2 related disease.

As used herein, the term “NO-related disease" refers to any pathology or a disease wherein NO expression, secretion, or both, initiates, propagates, involved, promotes, enhances, triggers, or any combination thereof, a pathological condition, or a disease or a symptom thereof. In some embodiments, NO-related disease is any disease or condition involving NO expression, secretion, or both, in the pathogenesis, pathophysiology, or both, of the disease or condition.

In some embodiments, at least one of: IL-6-related disease, IL-1β-related disease, and TNF-α-related disease, is selected from: endometriosis, dysmenorrhea, dyspareunia, rheumatoid arthritis, juvenile rheumatoid arthritis, systemic onset juvenile rheumatoid arthritis, psoriatic arthritis, ankylosing spondilitis, gastric ulcer, seronegative arthropathies, GYNI-P-002-IL asteoarthritis, inflammatory bowel disease, ulcerative colitis, systemic lupus erythematosis, antiphospholipid syndrome, iridocyclitis/uveitis/optic neuritis, idiopathic pulmonary fibrosis, systemic vasculitis/Wegener's granulomatosis, sarcoidosis, orchitis/vasectomy reversal procedures, allergic/atopic diseases, asthma, allergic rhinitis, eczema, allergic contact dermatitis, allergic conjunctivitis, hypersensitivity pneumonitis, transplants, organ transplant rejection, graft-versus-host disease, systemic inflammatory response syndrome, sepsis syndrome, gram positive sepsis, gram negative sepsis, culture negative sepsis, fungal sepsis, neutropenic fever, urosepsis, meningococcemia, trauma/hemorrhage, bums, ionizing radiation exposure, acute pancreatitis, adult respiratory distress syndrome, rheumatoid arthritis, alcohol-induced hepatitis, chronic inflammatory pathologies, sarcoidosis, Crohn's pathology, sickle cell anemia, diabetes, nephrosis, atopic diseases, hypersensitivity reactions, allergic rhinitis, hay fever, perennial rhinitis, conjunctivitis, asthma, urticaria, systemic anaphylaxis, dermatitis, pernicious anemia, hemolytic disease, thrombocytopenia, graft rejection of any organ or tissue, kidney transplant rejection, heart transplant rejection, liver transplant rejection, pancreas transplant rejection, lung transplant rejection, bone marrow transplant (BMT) rejection, skin allograft rejection, cartilage transplant rejection, bone graft rejection, small bowel transplant rejection, fetal thymus implant rejection, parathyroid transplant rejection, xenograft rejection of any organ or tissue, allograft rejection, anti-receptor hypersensitivity reactions, Grave’s disease, Raynaud's disease, type B insulin-resistant diabetes, asthma, myasthenia gravis, antibody meditated cytotoxicity, type III hypersensitivity reactions, systemic lupus erythematosus, POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes syndrome), polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, skin changes syndrome, antiphospholipid syndrome, pemphigus, scleroderma, mixed connective tissue disease, idiopathic Addison's disease, diabetes mellitus, chronic active hepatitis, primary biliary cirrhosis, vitiligo, vasculitis, post-M cardiotomy syndrome, type IV hypersensitivity, contact dermatitis, hypersensitivity pneumonitis, allograft rejection, granulomas due to intracellular organisms, drug sensitivity, metabolic/idiopathic, Wilson's disease, hemochromatosis, alpha-1-antitrypsin deficiency, diabetic retinopathy, Hashimoto's thyroiditis, osteoporosis, hypothalamic-pituitary-adrenal axis evaluation, primary biliary cirrhosis, thyroiditis, encephalomyelitis, cachexia, cystic fibrosis, neonatal GYNI-P-002-IL chronic lung disease, chronic obstructive pulmonary disease (COPD), familial hematophagocytic lymphohistiocytosis, dermatologic conditions, psoriasis, alopecia, nephrotic syndrome, nephritis, glomerular - nephritis, acute renal failure, hemodialysis, uremia, toxicity, preeclampsia, okt3 therapy, anti-cd3 therapy, cytokine therapy, chemotherapy, radiation therapy (e.g., including but not limited to asthenia, anemia, cachexia, and the like), chronic salicylate intoxication, sleep apnea, obesity, heart failure, sinusitis, and inflammatory bowel disease.

In some embodiments, at least one of: COX-related disease and NO-related disease, is selected from: endometriosis, dysmenorrhea, dyspareunia, rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, systemic lupus erythematosus, juvenile arthritis, asthma, bronchitis, menstrual cramps, tendinitis, bursitis, psoriasis, eczema, burns, dermatitis, inflammatory bowel disease, Crohn's disease, gastritis, irritable bowel syndrome, ulcerative colitis, cancer, such as colorectal cancer, vascular disease, migraine headaches, periarteritis nodosa, thyroiditis, aplastic anemia, Hodgkin's disease, sclerodoma, rheumatic fever, type I diabetes, myasthenia gravis, multiple sclerosis, sarcoidosis, nephrotic syndrome, Behcet's syndrome, polymyositis, gingivitis, hypersensitivity, swelling occurring after injury, myocardial ischemia, retinitis, retinopathies, conjunctivitis, uveitis, ocular photophobia, acute injury to the eye tissue, pulmonary inflammation, such as that associated with viral infections and cystic fibrosis, cortical dementias including Alzheimer's disease, allergic rhinitis, respiratory distress syndrome, endotoxin shock syndrome, atherosclerosis and central nervous system damage resulting from stroke, ischemia and trauma.

In some embodiments, treating comprises one or more of: increasing immune cell viability, reducing NO production, reducing COX expression, reducing cytokine secretion, or any combination thereof, in the subject.

In some embodiments, "reduce" or "reducing" is at least a: 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 100% reduction, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, "reduce" or "reducing" is by 5-100%, 5-90%, 5-80%, 5-70%, 5-60%, 5-50%, 5-40%, 5-30%, 5-25%, 5-20%, 5-10%, 10-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 20-100%, GYNI-P-002-IL -90%, 20-80%, 20-70%, 20-60%, 20-50%, 20-40%, 20-30%, 30-100%, 30-90%, 30-80%, 30-70%, 30-60%, 30-50%, 30-40%, 40-100%, 40-90%, 40-80%, 40-70%, 40-60%, 40-50%, 50-100%, 50-90%, 50-80%, 50-70%, 50-60%, 60-100%, 60-90%, 60-80%, 60-70%, 70-100%, 70-90%, 70-80%, 80-100%, 80-90%, or 90-100%. Each possibility represents a separate embodiment of the invention.

In some embodiments, "increase" or "increasing" is at least a: 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, or 1,000% increase, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, "increase" or "increasing" is by 5-10%, 5-50%, 5-100%, 10-50%, 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 50-200%, 50-300%, 50-400%, 50-500%, 100-200%, 100-300%, 100-400%, 100-500%, 100-600%, 100-700%, 100-800%, 100-900%, or 100-1000%. Each possibility represents a separate embodiment of the invention.

In some embodiments, "reduce" or "reducing" is compared to a control. In some embodiments, "increase" or "increasing" is compared to a control. In some embodiments, the control comprises a control subject. In some embodiments, the control subject comprises the subject in need thereof, prior treatment with the method disclosed herein. In some embodiments, the control subject comprises a subject afflicted with the same disease as the subject disclosed herein, not being treated with the method disclosed herein. In some embodiments, the control subject comprises a subject afflicted with the same disease as the subject disclosed herein, being treated with at least one of: an immunomodulatory therapy, an immunosuppressive therapy, a nonsteroidal anti-inflammatory drug (NSAID), an anti-hormonal drug, or any combination thereof.

The terms "reduce", "reducing", "inhibit" and "inhibiting" are used interchangeably.

Methods for determining immune cell viability, nitric oxide production, COX expression, and IL-6 secretion level, are common and would be apparent to one of ordinary skill in the art. Non-limiting examples for such methods, include, but are not limited to, immunoassays, such as enzyme-linked immunosorbent assay (ELISA), western blot, dot- GYNI-P-002-IL blot, MS-MS, RT-PCR, TUNEL apoptosis assay, trypan blue stain, acridine orange/ethidium bromide, real-time RT-PCR, or others, some of which are disclosed hereinbelow.

In some embodiments, the disease is selected from: an inflammatory disease, endometriosis, dysmenorrhea, or dyspareunia. In some embodiments, an inflammatory disease comprises or is an inflammatory lung disease or a viral-induced inflammatory disease.

In some embodiments, the disease is a women’s health related disease or condition.

As used herein, the phrase "women's health related disease or condition" comprises or consists of any health related disease or condition in women that results from, induced by, or involves inflammation, or any combination thereof.

In some embodiments, a women's health related disease is induced by inflammation in a woman's reproductive organ and/or a neighboring tissue thereto. In some embodiments, a neighboring tissue covers or underlies the reproductive organ. In some embodiments, a neighboring tissue comprises a skin tissue, a connective tissue, or both.

In some embodiments a woman's reproductive organ is selected from: breast, uterus, vulva, vagina, clitoris, ovary, cervix, and fallopian tube.

In some embodiments, a women's health related disease is induced by inflammation in the endometrium, myometrium, perimetrium, or any combination thereof. In some embodiments, a women's health related disease is induced by inflammation in the endometrium.

In some embodiments, a women's health related disease is induced by inflammation in a woman's urethra.

The phrases "induced by", "initiated by", "enhanced by", "propagated by", and "involves" are interchangeable.

In some embodiments, a women’s health related disease or condition is selected from: osteoporosis, vaginal atrophy and dryness, hypogonadism, skin atrophy, connective tissue disease, breast, endometrial, ovarian or uterine cancer, hot flashes, physical symptoms of menopause, or any combination thereof.

GYNI-P-002-IL In some embodiments, administering is topically administering, orally administering, or both.

As used herein, the terms “administering”, “administration”, and like terms refer to any method which, in sound medical practice, delivers a composition containing an active agent to a subject in such a manner as to provide a therapeutic effect. One aspect of the present subject matter provides for topical administration, oral administration, or both, of a therapeutically effective amount of a composition of the present subject matter to a patient in need thereof. Other suitable routes of administration can include parenteral, subcutaneous, intravenous, intramuscular, or intraperitoneal.

The term "therapeutically effective amount" refers to an amount of a drug effective to treat a disease or disorder in a mammal. The term “a therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result. The exact dosage form and regimen would be determined by the physician according to the patient's condition.

As used herein, the terms “treatment” or “treating” of a disease, disorder, or condition encompasses alleviation of at least one symptom thereof, a reduction in the severity thereof, or inhibition of the progression thereof. Treatment need not mean that the disease, disorder, or condition is totally cured. To be an effective treatment, a useful composition herein needs only to reduce the severity of a disease, disorder, or condition, reduce the severity of symptoms associated therewith, or provide improvement to a patient or subject’s quality of life.

As used herein, the term “prevention” of a disease, disorder, or condition encompasses the delay, prevention, suppression, or inhibition of the onset of a disease, disorder, or condition. As used in accordance with the presently described subject matter, the term "prevention" relates to a process of prophylaxis in which a subject is exposed to the presently described compositions or composition prior to the induction or onset of the disease/disorder process. This could be done where an individual has a genetic pedigree indicating a predisposition toward occurrence of the disease/disorder to be prevented. For example, this might be true of an individual whose ancestors show a predisposition toward certain types of, for example, inflammatory disorders. The term "suppression" is used to GYNI-P-002-IL describe a condition wherein the disease/disorder process has already begun but obvious symptoms of the condition have yet to be realized. Thus, the cells of an individual may have the disease/disorder, but no outside signs of the disease/disorder have yet been clinically recognized. In either case, the term prophylaxis can be applied to encompass both prevention and suppression. Conversely, the term "treatment" refers to the clinical application of active agents to combat an already existing condition whose clinical presentation has already been realized in a patient.

All scientific and technical terms used herein have meanings commonly used in the art unless otherwise specified. The definitions provided herein are to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

Before specific aspects and embodiments of the invention are described in detail, it is to be understood that this invention is not limited to particular methods, and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

In the discussion unless otherwise stated, adjectives such as “substantially” and “about” modifying a condition or relationship characteristic of a feature or features of an embodiment of the invention, are understood to mean that the condition or characteristic is defined to within tolerances that are acceptable for operation of the embodiment for an application for which it is intended. Unless otherwise indicated, the word “or” in the specification and claims is considered to be the inclusive “or” rather than the exclusive or, and indicates at least one of, or any combination of items it conjoins.

It should be understood that the terms “a” and “an” as used above and elsewhere herein refer to “one or more” of the enumerated components. It will be clear to one of ordinary skill in the art that the use of the singular includes the plural unless specifically stated otherwise. Therefore, the terms “a”, “an” and “at least one” are used interchangeably in this application.

GYNI-P-002-IL For purposes of better understanding the present teachings and in no way limiting the scope of the teachings, unless otherwise indicated, all numbers expressing quantities, percentages or proportions, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

In the description and claims of the present application, each of the verbs, “comprise”, “include” and “have” and conjugates thereof, are used to indicate that the object or objects of the verb are not necessarily a complete listing of components, elements or parts of the subject or subjects of the verb.

Other terms as used herein are meant to be defined by their well-known meanings in the art.

Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive.

Throughout this specification and claims, the word “comprise”, or variations such as “comprises” or “comprising,” indicate the inclusion of any recited integer or group of integers but not the exclusion of any other integer or group of integers.

As used herein, the term “consists essentially of”, or variations such as “consist essentially of” or “consisting essentially of” as used throughout the specification and claims, indicate the inclusion of any recited integer or group of integers, and the optional inclusion of any recited integer or group of integers that do not materially change the basic or novel properties of the specified method, structure or composition.

As used herein, the terms "comprises", "comprising", "containing", "having" and the like can mean "includes", "including", and the like; "consisting essentially of or "consists essentially" likewise has the meaning ascribed in U.S. patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel GYNI-P-002-IL characteristics of that which is recited is not changed by the presence of more than that which is recited, but excludes prior art embodiments. In one embodiment, the terms "comprises", "comprising", "having" are/is interchangeable with "consisting".

Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination of the following examples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.

EXAMPLES Generally, the nomenclature used herein, and the laboratory procedures utilized in the present invention include chemical, molecular, biochemical, and cell biology techniques. Such techniques are thoroughly explained in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed. (1994); "Cell Biology: A Laboratory Handbook", Volumes I-III Cellis, J. E., ed. (1994); The Organic Chemistry of Biological Pathways by John McMurry and Tadhg Begley (Roberts and Company, 2005); Organic Chemistry of Enzyme-Catalyzed Reactions by Richard Silverman (Academic Press, 2002); Organic Chemistry (6th Edition) by Leroy "Skip" G Wade; Organic Chemistry by T. W. Graham Solomons and, Craig Fryhle.

EXAMPLE The effect of cannabinoids and flavonoids on cyclooxygenase isoforms 1 and 2 (COXand COX2) expression in murine monocytes RAW 264.7 cells The enzymes cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) catalyze the conversion of arachidonic acid to prostaglandin (PG) H2, the precursor of PGs and thromboxane. These lipid mediators play important roles in inflammation and pain. Specifically, endometriotic lesions are known to express high levels of COX2 and COX2-derived prostaglandin E2 (PGE2), compared with the normal endometrium. Elevated COX-levels lead to high cell proliferation, low level of apoptosis, high invasion, angiogenesis, GYNI-P-002-IL endometriosis-related pain and infertility. Thus, the inventors initially evaluated the effect of mono-treatments with cannabinoids, terpenes, and a flavonoid, on COX1 and COXexpression in lipopolysaccharide (LPS)- activated macrophages. The murine cell line RAW 264.7 was used as a model to evaluate COX1 and COX2 expression in the cultured stimulated macrophages.

As provided in Figure 1, treatment of the LPS-stimulated RAW 264.7 cells with either tetrahydrocannabinol (THC), cannabidiol (CBD), or cannabigerol (CBG), strongly reduced the expression of both isoforms of COX with a higher reduction of COX-expression. Interestingly, when comparing between the CBD effect to the CBG effect, CBD was found to reduce both COX-1 and COX-2 levels, whereas CBG showed more selectivity towards COX-2 isoform. Interestingly, though the flavonoid apigenin induced COX-expression, it showed a very strong and highly specific reduction of the COX-2 isoform.

Examination of treatment of LPS- activated RAW 246.7 cells with different ratios of THC and THCA (Figure 2), revealed that the molar ratio 70:30 of THC to THCA was most effective in reducing COX expression, and particularly in selectivity toward COX-2.

EXAMPLE The i n - v i t r o effect of cannabinoids and flavonoids on pro-inflammatory cytokines and nitric oxide (NO) secretion Next the inventors examined the effect of different treatment combinations, each consisting between a single compound to 7 compounds, on the inflammatory activity, as manifested by secretion of the pro-inflammatory cytokines IL-6, IL-1β, and TNF-α, as well as NO production, by LPS-stimulated RAW 264.7 cells. Overall, twenty-seven combinations, including 1-4 cannabinoids/apigenin in 2 doses (4 µM or 8 µM for cannabinoids, and 2 µM or 4 µM for apigenin) were evaluated.

Interleukin (IL)-6 levels are known to be increased in peritoneal fluid (PF) of patients with endometriosis. Hyperactivated macrophages in the PF are thought to contribute to the pathogenesis of endometriosis by secreting growth factors and pro-inflammatory cytokines, mainly IL-6, IL-1β and tumor necrosis factor α (TNF-α). The inventors examined the effect of different treatment combinations, each consisting between a single compound GYNI-P-002-IL to 7 compounds, on the inflammatory activity, as manifested by secretion of the pro-inflammatory cytokines IL-6, IL-1β, and TNF-α, as well as NO production, by RAW 264.cells. Overall, twenty-seven combinations, including 1-4 cannabinoids/apigenin in 2 doses (4 µM or 8 µM for cannabinoids, and 2 µM or 4 µM for apigenin) were evaluated.

As shown in Figure 3, mono-treatments with one cannabinoid or flavonoid selected from: CBD, CBDA, CBG, THCV, and apigenin, had a moderate effect on IL-secretion, wherein the most profound effect among the mono-treatments was demonstrated for 4 µM apigenin, which reduced IL-6 secretion by 48% as compared to untreated LPS- stimulated cells. Surprisingly, all dual-treatments with one of the combinations: (i) 8 µM CBD and 4 µM apigenin, (ii) 8 µM CBDA and 8 µM CBG, and (iii) 8 µM CBG and 4 µM apigenin, reduced IL-6 secretion by more than 60% compared to untreated LPS- stimulated cells (63%, 79%, and 85%, respectively), whereas dual-treatments with 8 µM CBD and µM CBDA, or with 8 µM CBD and 8 µM CBG had a very moderate effect on IL-6 secretion (7% and 29% respectively). In fact, the combination of CBG and apigenin was found to synergistically affect IL-6 secretion, as compared to the CBG mono-treatment and to the apigenin mono-treatment. Moreover, the abovementioned dual-treatments with either CBD and apigenin, CBG and apigenin, or CBDA and apigenin, where much more potent, in the context of IL-6 secretion, as compared to a treatment with the three psychoactive compounds: 4µM THC, 4µM THCA and 8 µM THCV, which was found to reduce IL-secretion only by 18% (Figure 3).

The next step was to examine the different combinations of the cannabinoids and the flavonoid apigenin on IL-1β secretion by LPS- stimulated murine RAW 264.7 cells. As shown in Figure 4, a synergetic effect was observed for the dual-treatments with either CBD and apigenin, or with CBG and apigenin, which reduced IL-1β secretion by 84% and 57%, respectively.

Similar results were obtained when TNF-α secretion, by LPS-stimulated murine RAW 264.7 cells, was examined (Figure 5). All dual-treatments with one of: (i) CBD and apigenin, (ii) CBDA and CBG, and (iii) CBG and apigenin, had the most profound effect on GYNI-P-002-IL inhibition of TNF-α secretion (reduced TNF-α secretion by 85%, 45%, and 57%, respectively).

Examination of NO production by LPS- stimulated murine RAW 264.7 cells, treated with different cannabinoids and apigenin combinations revealed a similar trend (Figure 6), in which the dual-treatment with CBD and apigenin, or with CBDA and CBG, was more effective in reducing NO production as compared to the dual treatment with either CBD and CBDA, or with CBD and CBG. It was also surprisingly found that addition of a psychoactive compound has an ameliorating effect in the context of NO production, as the most powerful effect was observed for a combination treatment comprising the compounds CBD, CBDA, CBG, apigenin, THC, THCA, and THCV, which reduced NO production by 92%, as compared to LPS-stimulated untreated cells. These results indicate an increased anti-inflammatory effect for compositions comprising the cannabinoid CBD or CBG and the flavonoid apigenin, as well as to the composition comprising CBG and CBDA, as compared to a composition comprising cannabinoid and its perspective acid precursor (e.g., CBD and CBDA), in inflammatory diseases mediated by macrophages activation, pro-inflammatory cytokine secretion, NO production and COX induction, such as endometriosis, and its related symptoms.

While the present invention has been particularly described, persons skilled in the art will appreciate that many variations and modifications can be made. Therefore, the invention is not to be construed as restricted to the particularly described embodiments, and the scope and concept of the invention will be more readily understood by reference to the claims, which follow.

Claims (25)

GYNI-P-002-IL CLAIMS What is claimed is:

1. A pharmaceutical composition comprising: a. a cannabinoid selected from cannabidiol (CBD) or cannabigerol (CBG); and b. apigenin, wherein said CBD or CBG and said apigenin are present in said pharmaceutical composition in a weight per weight (w/w) or mole per mole (m:m) ratio ranging from 1:1 to 10:1.

2. The pharmaceutical composition of claim 1, wherein said CBD or CBG and apigenin are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 4:1.

3. The pharmaceutical composition of claim 1 or 2, further comprising cannabidiolic acid (CBDA).

4. The pharmaceutical composition of claim 3, wherein said CBD or CBG and said CBDA are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 2:1 to 1:2.

5. The pharmaceutical composition of any one of claims 1 to 4, wherein any one of: (i) said cannabinoid is CBD, and said pharmaceutical composition further comprises CBG; and (ii) said cannabinoid is CBG, and said pharmaceutical composition further comprises CBD.

6. The pharmaceutical composition of claim 5, wherein said CBD and said CBG are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 2:1 to 1:2.

7. The pharmaceutical composition of any one of claims 1 to 6, further comprising an additional compound selected from the group consisting of: tetrahydrocannabinol (THC), tetrahydrocannabinol acid (THCA), tetrahydrocannabivarin (THCV), and any combination thereof. GYNI-P-002-IL

8. The pharmaceutical composition of claim 7, wherein said CBD or CBG and said additional compound are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 4:1.

9. The pharmaceutical composition of claim 7 or 8, wherein said THC and said THCA are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 2:1 to 2.5:1.

10. The pharmaceutical composition of any one of claims 1 to 9, wherein said cannabinoid is CBG.

11. A pharmaceutical composition comprising an active ingredient and a carrier, wherein said active ingredient consists essentially of: (i) a cannabinoid selected from CBD and CBG; and (ii) apigenin.

12. The pharmaceutical composition of claim 11, wherein said CBD or CBG and said apigenin are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 10:1.

13. The pharmaceutical composition of claim 11 or 12, further comprising CBDA.

14. The pharmaceutical composition of claim 13, wherein said CBD or CBG and said CBDA are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 2:1 to 1:2.

15. The pharmaceutical composition of any one of claims 11 to 14, further comprising an additional compound selected from the group consisting of: THC, THCA, and THCV.

16. The pharmaceutical composition of claim 15, wherein said CBD or CBG and said additional compound is present in said pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 4:1.

17. The pharmaceutical composition of any one of claims 11 to 16, wherein said cannabinoid is CBG. GYNI-P-002-IL

18. A pharmaceutical composition comprising a first active ingredient and a carrier, wherein said first active ingredient consists essentially of CBDA and CBG.

19. The pharmaceutical composition of claim 18, wherein said CBDA and said CBG are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 1:10 to 10:1.

20. The pharmaceutical composition of claim 18 or 19, further comprising a second active ingredient, wherein said second active ingredient consists essentially of apigenin.

21. The pharmaceutical composition of any one of claims 18 to 20, wherein said CBDA and said CBG are present in said composition in a w/w or m:m ratio ranging from 1:2 to 2:1.

22. The pharmaceutical composition of claim 20 or 21, wherein said CBDA or CBG and said apigenin are present in said pharmaceutical composition in a w/w or m:m ratio ranging from 1:1 to 10:1.

23. The pharmaceutical composition of any one of claims 1 to 22, for use in the treatment or prevention of an interleukin-6 (IL-6)-related disease, an IL-1β- related disease, a tumor necrosis factor α (TNF-α)-related disease, a cyclooxygenase (COX)-related disease, a nitric oxide (NO)-related disease, or any combination thereof.

24. The pharmaceutical composition for use according to claim 23, wherein said disease is selected from the group consisting of: an autoimmune disease, an inflammatory disease, endometriosis, dysmenorrhea, and dyspareunia.

25. The pharmaceutical composition for use according to claim 23 or 24, wherein said COX is COX2.