Cannabidiol compositions for use in treating cardiac conditions

Abstract

A pharmaceutical composition containing an effective amount of cannabidiol (CBD) for use in the treatment or prevention of cardiac conditions, including heart failure, acute myocarditis, toxicity caused by anticancer therapy, acute pericarditis, cardiac sarcoidosis, inflammatory cardiomyopathy, and atherosclerosis, as well as related uses and methods. The present disclosure demonstrates that CBD is effective in (i) reducing cardiac hypertrophy; (ii) reducing cardiac fibrosis; (iii) reducing BNP levels; (iv) reducing levels of the cytokine IL1β; (v) reducing levels of the cytokine IL6; (vi) reducing levels of CD69; (vii) increasing levels of the cytokine IL10; and (viii) combinations thereof. The composition is preferably adapted for parenteral administration.

Description

[Technical Field]

[0001]

[0001] This application claims priority from U.S. Provisional Patent Application No. 62 / 926,066, filed October 25, 2019, the entire contents of which are incorporated herein by reference.

[0002]

[0002] Field

[0003] The present invention relates generally to pharmaceutical compositions containing cannabidiol (CBD) and the treatment or prevention of diseases and disorders using such compositions. [Background technology]

[0003]

[0004] background

[0005] Chronic heart failure (CHF) affects over 26 million people worldwide. Over 6 million adults in Canada and the United States suffer from chronic heart failure, and it remains the leading cause of death and hospitalization, with associated medical costs exceeding $30 billion annually in the United States alone.

[0004]

[0006] People with heart failure (HF) suffer from shortness of breath, rapid heart rate, edema, and decreased exercise capacity. They often struggle with simple daily activities and are frequently hospitalized. For many people, these symptoms significantly reduce their quality of life.

[0005]

[0007] Heart failure occurs when the heart is unable to pump enough blood to meet the body's needs. There are two types of heart failure: heart failure with reduced ejection fraction (HFrEF, also called systolic heart failure) and heart failure with preserved ejection fraction (HFpEF, formerly called diastolic heart failure). In heart failure with reduced ejection fraction (HFrEF), there is reduced contractility of the left ventricle such that not enough blood is pumped into the circulation with each contraction of the heart. In heart failure with preserved ejection fraction (HFpEF), the problem is primarily limited filling of the left ventricle during relaxation. Lesions cause the left ventricle to stiffen, and it does not relax normally. As a result, it cannot fill normally, and pressure begins to increase in the left heart chamber and lungs. Increased pressure in the lungs causes shortness of breath.

[0006]

[0008] HFpEF is a complex syndrome. The pathophysiological mechanisms are still incompletely understood, which is one of the reasons why there is no effective treatment to date. However, several pathological conditions are recognized to be frequently associated with HFpEF. Important among these are components of metabolic syndrome (impaired glucose tolerance, obesity, hypertension, and dyslipidemia). Of particular importance is obesity, which leads to a systemic inflammatory state and contributes to cardiac inflammation, which is associated with fibrosis and reduced ventricular compliance (relaxation). Hypertension (HTN) is also a major precursor to HFpEF and leads to increased left ventricular (LV) muscle thickness (LV hypertrophy) and reduced LV compliance. This inability of the left ventricle to fill normally is also referred to as left ventricular diastolic dysfunction (LVDD). Some consider HTN to be the most significant risk factor for the development of LVDD.

[0007]

[0009] The publication Glezeva et al., "Role of inflammation in the pathogenesis of heart failure with preserved ejection fraction and its potential as a therapeutic target," (Heart Fail Rev (2014) 19:681-694; DOI 10.1007 / s10741-013-9405-8), incorporated herein by reference, describes research showing that inflammation is an early and underlying trigger of ventricular remodeling in HTN and contributes to LVDD. Research suggests that inflammation is caused by elevated levels of endothelial adhesion molecules and increased production and release of inflammatory cytokines and chemokines within the tissue. The latter promotes the infiltration of activated inflammatory cells, particularly monocytes, into cardiac tissue. Increased monocyte infiltration is seen in the early and late stages of HTN and HFpEF. Once inside the tissue, monocytes are thought to differentiate into macrophages, promoting cardiac inflammation, tissue injury, and myocardial fibrosis. This mechanism is thought to be part of the progression to HFpEF. The authors conclude that myocardial inflammation plays a role in the pathophysiology of HFpEF and suggest that therapeutic approaches that intervene in inflammatory pathways should be utilized in addition to antihypertensive treatment in at-risk patients or those with LVDD and / or HFpEF. The authors conclude that a better understanding of the ongoing inflammatory response in HTN and LVDD, as well as more large-scale randomized clinical trials in patients with HFpEF, are needed.

[0008]

[0010] In addition to hypertension, several risk factors are associated with the development of HFpEF. The first is diabetes, a common risk factor for not only heart failure but also coronary atherosclerosis, the usual cause of heart attacks. Other major risk factors are obesity and aging. Interestingly, aging, diabetes, and obesity are all conditions associated with increased inflammation throughout the body. Adipocytes are the primary producers of inflammatory cytokines that circulate to other tissues. The combination of hypertension, diabetes, and obesity is extremely common in HFpEF, and each contributes to the inflammatory environment that many believe is the cause of this syndrome.

[0009]

[0011] Cannabinoids are compounds naturally occurring in the plant Cannabis sativa. The major cannabinoid components are cannabidiol (CBD) and tetrahydrocannabinol (THC). THC is known to increase pulse rate, cause conjunctival redness, and exert antipsychotic (mood-altering) effects, such as euphoria. In contrast to THC, CBD is non-addictive. Therefore, CBD is more commonly used as a medicinal ingredient to treat a wide range of conditions, including epilepsy, chronic pain, anxiety, and insomnia. The anti-inflammatory effects of CBD have been noted in several publications. See, for example, International Publication No. WO 2014 / 117999 A1 (chronic inflammation and inflammatory diseases), International Publication No. WO 2017 / 191630 A1 (liver inflammation), U.S. Patent No. 9,549,906 (ocular inflammation), and International Publication No. WO 2018148152 (neuroinflammation).

[0010]

[0012] Cannabidiol (CBD) is lipid-soluble and virtually water-insoluble. Furthermore, it is prone to inactivation in the liver due to first-pass metabolism. This high first-pass metabolism results in low effective blood levels and an overall bioavailability of less than 10% when taken orally. Summary of the Invention [Problem to be solved by the invention]

[0011]

[0013] There is a continuing need to provide improved compositions containing CBD that avoid first-pass hepatic metabolism to optimize and maintain blood levels of the drug. There is also a continuing need to better understand the pathophysiological mechanisms of cardiac conditions, including heart failure, to provide new therapies for treating and / or preventing the same. [Means for solving the problem]

[0012]

[0014] overview

[0015] The present inventors have found that parenterally, e.g., subcutaneously administered CBD is effective in reducing cardiac hypertrophy and myocardial fibrosis and beneficially modulating the levels of molecular markers of remodeling and inflammation in an in vivo non-ischemic (hypertension-induced) mouse model of heart failure. Surprisingly, these effects were observed at very low doses. The present inventors also found that CBD is effective in preventing cell enlargement (hypertrophy) in an in vitro model of cardiomyocyte hypertrophy using H9c2 cells. Furthermore, the present inventors have conducted studies showing that CBD is effective in reducing the expression of CD69 (a marker of inflammation) by human lymphocytes and monocytes in vitro. These findings, and those in the published literature, support a beneficial role for CBD in the treatment or prevention of cardiac conditions, including heart failure (e.g., HFpEF), acute myocarditis, toxicity caused by certain anti-cancer therapies (e.g., doxorubin, checkpoint inhibitors), acute pericarditis, cardiac sarcoidosis, certain dilated cardiomyopathies (inflammatory cardiomyopathies), and even atherosclerosis (coronary artery disease), each of which has inflammation as a prominent component and involves varying degrees of hypertrophy or fibrosis.

[0013]

[0016] Thus, according to a first aspect, the present invention provides the use of an effective amount of CBD for treating or preventing a cardiac condition selected from the group consisting of heart failure (e.g., HFpEF), acute myocarditis, toxicity caused by anti-cancer therapy (e.g., doxorubicin, checkpoint inhibitors), acute pericarditis, cardiac sarcoidosis, inflammatory cardiomyopathy, and atherosclerosis.

[0014]

[0017] In certain embodiments, the use of CBD may be associated with the following effects in a subject: (a) Reduces cardiac hypertrophy; (b) whether it reduces cardiac fibrosis; (c) whether it reduces BNP levels; (d) whether it reduces levels of the cytokine IL1β; (e) whether it reduces IL6 levels; (f) whether it reduces the level of CD69; (g) increasing the level of the cytokine IL10; or (h) resulting in any combination of the above It is effective for this purpose.

[0015]

[0018] Those skilled in the art will recognize that if a reduction in blood levels of BNP is observed, cardiac function, including cardiac dilation, will improve in subjects with heart failure. Furthermore, if the levels of cytokines IL1β, IL6, and CD69 are reduced or the level of cytokine IL10 is increased, this indicates a reduction in inflammation, which is known to underlie the above-mentioned inflammatory cardiac pathologies.

[0016]

[0019] In selected embodiments, CBD is used to effect at least two, three, four, five, six, or all of (a)-(g) above in a subject.

[0017]

[0020] According to a second aspect of the present invention, CBD can be present in a parenteral composition, which comprises, consists essentially of, or consists of an effective amount of CBD and an effective amount of at least one pharmaceutically acceptable solvent for solubilizing the CBD in the composition. Parenteral administration of CBD will avoid first-pass liver metabolism and thus optimize and maintain blood levels of the drug. The composition is adapted to be administered by any parenteral route, including intramuscular (IM), intravenous (IV), intraperitoneal (IP), and subcutaneous (SC) routes of injection. profit Preferably, the composition is adapted to be administered by SC and IM injection.

[0018]

[0021] The present invention also contemplates the use of CBD in combination with one or more additional anti-inflammatory pharmaceutically active agents, such as beta-caryophyllene (BCP), methotrexate (MTX), and cyclosporine (CsA). These other agents may be administered simultaneously or sequentially with CBD, or (depending on the agent) may be administered in the same composition with CBD. BCP and CsA are agents that can be included in the same composition as CBD. In some embodiments, the pharmaceutically active agent in the composition consists of CBD. In other embodiments, the pharmaceutically active agent consists of CBD and BCP. Furthermore, the present uses, compositions, and methods may complement existing therapies and methods for treating cardiac conditions and improving patient outcomes.

[0019]

[0022] The at least one pharmaceutically acceptable solvent may be selected from the group consisting of natural and synthetic medium-chain (C6-C12) triglycerides (MCTs), structured lipids, propylene glycol di(caprylate / caprate), vegetable oils, N-methyl-2-pyrrolidone, polyoxyethylene castor oil derivatives, dimethylacetamide (DMA), ethanol, glycerin, PEG 300, PEG 400, polyoxyethylene 20 sorbitan monooleate, propylene glycol (PG), polyglycol mono- and diesters of 12-hydroxystearic acid, solvents sold under the trademark Kolliphor™, and mixtures thereof. These solvents are useful for solubilizing lipophilic compounds such as CBD, BCP, and CsA. On the other hand, other (hydrophilic) solvents may be used for non-lipophilic MTX.

[0020]

[0023] In some embodiments, the at least one pharmaceutically acceptable solvent is selected from the group consisting of natural and synthetic medium-chain (C6-C12) triglycerides (MCTs). Further, at least about 95% of the MCTs may consist of C8 triglycerides, C10 triglycerides, or mixtures thereof.

[0021]

[0024] The pharmaceutically active agents are present in an "effective amount," as defined below. In embodiments in which the pharmaceutically active agents consist of CBD or CBD and BCP, each pharmaceutically active agent may be present in an amount of at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 mg / mL of composition. Alternatively, or additionally, each pharmaceutically active agent may be present in an amount up to about 350, 300, 250, 200, or 150 mg / mL of composition. Typically, about 0.1, 0.5, 1, 1.5, 2, 3, or 4 to about 30, 25, 20, 15, 10, 9, 8, 7, 6, or 5 mg of each pharmaceutically active agent is administered per kg of body weight. Preferably, each pharmaceutically active agent is administered in an amount of about 0.1 to about 10 mg / kg of body weight, or about 1 to about 10 mg / kg of body weight.

[0022]

[0025] In embodiments in which the pharmaceutically active agent comprises CBD and BCP, the weight ratio of CBD to BCP can vary, provided that each active agent is present in an effective amount. For example, the weight ratio of CBD to BCP can be from about 1:4 to about 4:1, from about 1:2 to about 2:1, or about 1:1.

[0023]

[0026] In some embodiments, the parenteral composition is substantially free of water. In the same or other embodiments, the composition is substantially free of micelles.

[0024]

[0027] According to a third aspect, the present invention provides a method for treating and / or preventing a cardiac condition selected from the group consisting of heart failure (e.g., HFpEF), acute myocarditis, toxicity caused by anti-cancer therapy (e.g., doxorubin, checkpoint inhibitors), acute pericarditis, cardiac sarcoidosis, inflammatory cardiomyopathy, and atherosclerosis. The method comprises: (i) identifying a subject having or at risk of having said cardiac condition; and (ii) administering to the subject an effective amount of CBD; and administering CBD to the subject, (a) Reduces cardiac hypertrophy; (b) whether it reduces cardiac fibrosis; (c) whether it reduces BNP levels; (d) reducing levels of the cytokine IL1?; (e) whether it reduces IL6 levels; (f) whether it reduces the level of CD69; (g) increasing the level of the cytokine IL10; or (h) resulting in any combination of the above.

[0025]

[0028] In selected embodiments, administration of CBD results in at least two, three, four, five, six, or all of (a)-(g) above in a subject.

[0026]

[0029] Preferably, administration is by a parenteral route, which avoids first-pass hepatic metabolism. CBD may therefore be present in a parenteral composition according to the second aspect of the invention.

[0027]

[0030] In methods according to the invention, the composition can be administered at least 1, 2, or 3 times per week, at least once daily, or at least twice daily for 1, 2, 3, 4, 5, 6, 7, 8 weeks or more. When treating chronic conditions, the duration of treatment can be indefinite, and it is contemplated that the amount of pharmaceutically active ingredient is adjusted to a dosage that avoids long-term toxicity. Depending on the frequency of administration, each dose of the parenteral composition can contain from about 1, 2, 3, 4, or 5 to about 20, 15, or 10 mg of CBD (or each of CBD and BCP) per kg of body weight.

[0028]

[0031] Subjects at risk for developing heart failure (HFpEF) include older subjects (e.g., over 60 years old) and those with hypertension, diabetes, and / or obesity. Regarding acute myocarditis, this is a post-viral condition, and the affected population may be younger individuals. Cardiac sarcoidosis is uncommon overall, but is more prevalent in African Americans, usually in the 30-40 age group. It is characterized by the presence of granulomas in cardiac tissue, and diagnosis may require endomyocardial biopsy. Inflammatory cardiomyopathy often occurs in young adults and leads to chronic heart failure. In some cases, it may be the end result of acute myocarditis. More and more cancer patients are taking medications (such as anthracyclines and checkpoint inhibitors), which can affect the heart and cause inflammation-induced damage. In some cases, for example, after taking checkpoint inhibitors, the condition can rapidly worsen, leading to cardiogenic shock or death. Acute pericarditis is an acute inflammation of the pericardium and can include various inflammatory damage to the adjacent heart. It tends to occur in younger populations but can occur at any age. Atherosclerotic heart disease is a common cause of myocardial infarction and is usually associated with coronary thrombosis. The pathogenesis of thrombosis is widely believed to result, at least in part, from an inflammatory process in the arterial wall.

[0029]

[0032] BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The invention may be better understood with reference to the following description in conjunction with the following drawings. [Brief explanation of the drawings]

[0030] [Figure 1]

[0034] 1 includes selected confocal microscopy images showing the effect of administering different doses of CBD on the cell surface area of ​​H9c2 cells in an in vitro model of cardiomyocyte hypertrophy. [Figure 2]

[0035] 1 is a graph showing the effect of administration of different doses of CBD on the cell surface area of ​​H9c2 cells in an in vitro model of cardiomyocyte hypertrophy. [Figure 3a]

[0036] 1 is a graph showing the reduction in the level of mRNA expression of B-type natriuretic peptide (BNP), a remodeling biomarker, by CBD administered at different doses in an in vitro model of cardiomyocyte hypertrophy. [Figure 3b]

[0036] A graph showing the reduction in collagen mRNA expression levels by CBD administered at different doses in an in vitro model of cardiomyocyte hypertrophy. [Figure 4]

[0037] 1 is a graph showing the reduction of mitochondrial reactive oxygen species (MitoSOX), a marker of inflammation, by CBD administered at different doses in an in vitro model of cardiomyocyte hypertrophy. [Figure 5]

[0038] 1 depicts a depiction of the experimental design used to measure the effects of CBD administration in a mouse model of non-ischemic heart failure. [Figure 6A]

[0039] Figure 5 shows a representative slide of mouse heart tissue stained with Masson's Trichrome stain, demonstrating reduced fibrosis (reduced blue staining) after administration of CBD from the experiment. [Figure 6B]

[0039] Figure 5 shows a representative slide of mouse heart tissue stained with Masson's Trichrome stain, demonstrating reduced fibrosis (reduced blue staining) after administration of CBD from the experiment. [Figure 6C]

[0039] Figure 5 shows a representative slide of mouse heart tissue stained with Masson's Trichrome stain, demonstrating reduced fibrosis (reduced blue staining) after administration of CBD from the experiment. [Figure 6D]

[0039] Figure 5 shows a representative slide of mouse heart tissue stained with Masson's Trichrome stain, demonstrating reduced fibrosis (reduced blue staining) after administration of CBD from the experiment. [Figure 7]

[0040] 6 is a graph showing the reduction in fibrosis after CBD administration from the experiment of FIG. 5. [Figure 8]

[0041] 6 is a graph showing the reduction in muscle cell area (hypertrophy) following CBD administration for the experiment of FIG. 5. [Figure 9]

[0042] 6 is a graph showing the reduction in the level of BNP mRNA expression after CBD administration from the experiment of FIG. 5. [Figure 10a]

[0043] FIG. 6 is a graph showing the levels of mRNA expression of the inflammatory cytokine IL1β after CBD administration in the experiment of FIG. 5. [Figure 10b]

[0043] A graph showing the level of mRNA expression of the inflammatory cytokine IL6 after CBD administration in the experiment of Figure 5. [Figure 10c]

[0043] A graph showing the level of mRNA expression of the anti-inflammatory cytokine IL10 after CBD administration in the experiment of Figure 5. [Figure 11]

[0044] FIG. 10 is a graph showing the effect of CBD administration on CD69 expression by stimulated / activated monocytes and lymphocytes (white blood cells) in another experiment. DETAILED DESCRIPTION OF THE INVENTION

[0031]

[0045] In the figure, the "p-value" is * or #=≦0.05, ** or ##=≦0.01 and *** or ###=≦0.001. The lower the "p-value", the higher the significance between groups.

[0032]

[0046] Detailed Description

[0047] definition

[0048] For clarity and to avoid ambiguity, certain terms are defined herein as follows:

[0033]

[0049] The term "pharmaceutically active agent" refers to any composition (e.g., drug, compound, or component) capable of providing a therapeutic effect in a subject (e.g., curing, alleviating, or preventing a disease or its symptoms), including drugs, cells, DNA, RNA, oligonucleotides, proteins, and peptides.

[0034]

[0050] When a composition, e.g., a compound or component, is described as having "X% purity," this means that one or more impurities may be present in an amount of from 100 to X% by weight, based on the total weight of the composition. The purity of a component can be determined by high performance liquid chromatography (HPLC) or other suitable means.

[0035]

[0051] The term "subject" means members of the animal kingdom, including humans and other mammals.

[0036]

[0052] As used herein, the term "treatment" is intended to mean halting or slowing the progression of a condition, disorder, or disease. The term "prophylaxis" is intended to mean preventing or delaying the onset of a condition, disorder, or disease. This term is intended to encompass "improving the quality of life," "prolonging the lifespan," and "improving the clinical outcome" of a subject suffering from or at risk of suffering from a condition, disorder, or disease, and does not necessarily mean "curing" the condition, disorder, or disease.

[0037]

[0053] As used herein, a "pharmaceutically acceptable excipient" means any substance that can be formulated with or present together with a pharmaceutically active agent to achieve a desired function or functions. To be "pharmaceutically acceptable," an excipient must: Mode of administration In consideration of the above, it must be non-toxic, safe for humans and animals, and compatible with the other ingredients in the composition. One of ordinary skill in the art, given the teachings herein and information in the public domain, will recognize which compounds or ingredients qualify as pharmaceutically acceptable excipients.

[0038]

[0054] "Pharmaceutically acceptable solvent" means a pharmaceutically acceptable excipient that is effective (either alone or in combination with other pharmaceutically acceptable excipients) to solubilize at least one pharmaceutically active agent in the overall composition.

[0039]

[0055] The phrases "at least one," "one or more," and "and / or" are open-ended phrases that are both conjunctive and disjunctive in operation. For example, each of the phrases "at least one of A, B, and C," "at least one of A, B, or C," "one or more of A, B, and C," "one or more of A, B, or C," and "A, B, and / or C" means A only, B only, C only, A and B together, A and C together, B and C together, or A, B, and C together.

[0040]

[0056] The term "a" or "an" entity refers to one or more of that entity. Thus, the terms "a" (or "an"), "one or more," and "at least one" may be used interchangeably herein. It should be noted that the term "or" is generally utilized in the sense of "and / or" unless the context clearly dictates otherwise.

[0041]

[0057] The term "comprising" means "including but not limited to." Thus, a composition that includes a list of ingredients may contain additional ingredients not expressly listed. It should also be noted that the terms "comprising," "including," and "having" can be used interchangeably.

[0042]

[0058] The term "consisting of" means "including the listed ingredients and additional ingredients that may be present in the listed ingredients as natural or commercially available impurities or additives." Natural and commercially available impurities and additives will be apparent to one of ordinary skill in the art. Synthetic cannabidiol (CBD) may contain up to about 0.5% w / w impurities, such as residual solvents and by-products of the manufacturing process. Thus, a composition "consisting of synthetic CBD" means a composition having at least about 99.5% w / w CBD and up to about 0.5% w / w impurities.

[0043]

[0059] The term "consisting essentially of" means "including the recited ingredients and any additional ingredients that do not materially affect the basic and novel properties of the present invention." The "basic and novel properties" refer to the usefulness of the composition in treating or preventing cardiac conditions as set forth herein. In embodiments in which CBD is utilized, the "basic and novel properties" refer to the stability and solubility of CBD in the composition, as well as the Administration It also means the suitability of the composition for:

[0044]

[0060] Unless otherwise specified, the terms "weight percent," "% w / w," "percent by weight," "wt. %," "wt. %," and variations thereof refer to the amount of a substance calculated by dividing the weight of the substance by the total weight of the composition containing that substance, multiplied by 100.

[0045]

[0061] Unless otherwise specified, the terms "volume percent," "vol. %," "percent by volume," "volume %," % v / v and variations thereof refer to the amount of a substance calculated by dividing the volume of the substance by the total volume of the composition containing that substance, multiplied by 100.

[0046]

[0062] The term "about" refers to variations in the expressed quantity that may occur due, for example, to measurements and liquid handling procedures used in preparing a pharmaceutical composition, differences in the manufacture, source, or purity of the ingredients used to prepare the composition, and / or differences in equilibrium conditions or reaction levels of the ingredients in the composition resulting from the initial mixture. For clarity, the term "about" includes variations of up to ±5% of the expressed value. Whether or not a value is modified by the term "about," the claims include equivalents to the value.

[0047]

[0063] As used herein, the term "effective amount" refers to an amount that will produce a desired effect based on the known purpose and function of the components associated with the present invention. For example, an effective amount of a pharmaceutically active agent is an amount that will be effective to provide the therapeutic effect described herein. An effective amount of a solvent is an amount that will be effective, alone or together with other components, to solubilize other or remaining components of the composition. What constitutes an effective amount can be determined by one of ordinary skill in the art through routine experimentation and in light of the teachings herein.

[0048]

[0064] As used herein, the expression "substantially free of Y" means that "Y" is not intentionally added, but may be present as an impurity or due to other factors. For example, substantially free of water. composition In this case, the composition may contain trace amounts of water due to the atmosphere and water present in the composition or its components exposed to the atmosphere. For clarity, a composition that is "substantially free of Y" contains no Y or up to 0.5% w / w "Y" of the composition.

[0049]

[0065] Values ​​recited herein are intended to include all values ​​meeting the stated parameters, including those not explicitly recited. Thus, for example, a value less than 1.0% w / w is intended to include less than 0.99% w / w, less than 0.98% wt.%, less than 0.97% wt.%, less than 0.90% w / w, less than 0.84% ​​w / w, less than 0.56% w / w, less than 0.01% w / w, etc. Thus, all ranges disclosed herein should be understood to encompass all subranges contained therein. For example, a stated range of "1 to 10" should be considered to include all subranges from a minimum value of 1 to a maximum value of 10 (inclusive), such as 1 to 6.3, or 5.5 to 10, or 2.7 to 6.1, etc.

[0050]

[0066] This specification contemplates the possibility of omitting any ingredient, even if that ingredient is not expressly listed herein as being included or excluded.

[0051]

[0067] The chemical structures herein are depicted according to conventional standards known in the art. Therefore, if an atom, such as a depicted carbon atom, appears to have an unsatisfied valence, the valence is considered to be satisfied by a hydrogen atom, even if the hydrogen atom is not necessarily explicitly depicted. Some structures of the compounds herein contain carbon atoms that give rise to stereoisomerism. That is, any chiral carbon center can be either (R)-stereochemistry or (S)-stereochemistry. It should be understood that isomers arising from such asymmetry (e.g., all enantiomers and diastereomers) are included within the scope of the present specification, provided that such isomers are known to possess pharmaceutical activity, as would be understood by one of ordinary skill in the art. Such isomers can be obtained in substantially pure form by conventional separation techniques and stereochemically controlled synthesis. Furthermore, alkenes can include either the E- or Z-configuration, where appropriate. The appropriate configuration is that which will provide the desired pharmaceutical activity.

[0052]

[0068] Cannabidiol (CBD)

[0069] The compositions contain (at least) cannabidiol (CBD) as the pharmaceutically active agent. The terms cannabidiol and CBD are used interchangeably herein to refer to a compound having the following chemical structure: [ka]

[0053]

[0070] CBD can be of natural or synthetic origin and can be in crystalline or oil form. All commercial sources of CBD are useful in the context of the present invention.

[0054]

[0071] Preferably, the CBD has a purity of at least 99.5%, 99.6%, 99.7%, 99.8% or 99.9%.

[0055]

[0072] A "synthetic cannabinoid" is a compound that has a cannabinoid-like structure and is produced using chemical or biosynthetic means. Methods for producing synthetic cannabidiol are known in the art. For example, CBD from NORAMCO, INC., headquartered in Wilmington, Delaware, USA, is produced according to processes such as those described in U.S. Patent Application Publication No. 2017 / 0008868A1 (issued as U.S. Patent No. 10,059,683) and U.S. Patent Application Publication No. 2018 / 031,976A1, both of which are incorporated herein by reference. Synthetic CBD produced by other processes and manufacturers can also be used to produce the present compositions, provided that it has the desired purity.

[0056]

[0073] Plant-derived CBD can be derived from various cannabis plants, including hemp, and can be purified using conventional means.

[0057]

[0074] The terms tetrahydrocannabinol, THC, delta-9-tetrahydrocannabinol and delta-9-THC are used interchangeably herein to refer to the chemical compound having the structure shown below. The term is used broadly herein to include double bond isomers and their stereoisomers. [ka]

[0058]

[0075] Those skilled in the art will recognize that commercial sources of CBD may contain trace amounts of impurities, including THC. For example, synthetic CBD may contain small amounts of residual solvents (e.g., methanol, n-heptane, dichloromethane, and triethylamine) and manufacturing by-products such as olivetol, monobromo-CBD, and delta-9-THC. Plant-derived CBD may contain small amounts of other cannabinoids (including THC), terpenes, and solvents or ingredients used in the purification process.

[0059]

[0076] In some embodiments, the compositions are "substantially free" of THC, meaning that THC is absent or present in any amount less than 0.5% w / w of the composition. In these embodiments, THC may be present in an amount less than 0.4, 0.3, 0.2, or 0.1% w / w of the composition. In the same or other embodiments, THC may be present in an amount less than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 ppm of the composition.

[0060]

[0077] In some embodiments, the concentration of CBD in the composition can range from about 10, 20, 30, 40, 50, 60, 70, 80, or 90 to about 350, 300, 250, 200, 150, or 100 mg / mL of composition.

[0061]

[0078] β-caryophyllene (BCP)

[0079] The compositions of the present invention may (optionally) contain β-caryophyllene (BCP) and / or its derivatives as an additional active pharmaceutical ingredient. BCP is also known as trans-(1R,9S)-8-methylene-4,11,11-trimethylbicyclo[7.2.0]undec-4-ene or [1R-(1R,4E,9S)]-4,11,11-trimethyl-8-methylene-bicyclo[7.2.0]undec-4-ene and its derivatives. BCP is a naturally occurring bicyclic sesquiterpene compound present in plant extracts of plants, including Cannabis sativa. It is a component of many essential oils, particularly clove (Syzygium aromaticum) oil, and is "Generally Recognized as Safe" (GRAS).

[0062]

[0080] It is predicted or expected that the anti-inflammatory activity of CBD will be synergistically enhanced by the addition of BCP. Caryophyllene is the only terpene known to interact with the endocannabinoid system (at the CB2 receptor). β-Caryophyllene selectively binds to the CB2 receptor and is a functional CB2 agonist. Furthermore, β-caryophyllene has been identified as a functional non-psychoactive CB2 receptor ligand in foodstuffs and as a macrocyclic anti-inflammatory cannabinoid in cannabis (Proc Natl Acad Sci USA. 2008 Jul. 1; 105 (26): 9099-104. doi: 10.1073 / pnas.0803601105. Epub 2008 Jun. 23. β-Caryophyllene is also a dietary cannabinoid).

[0063]

[0081] The weight ratio of BCP and its derivatives to CBD can be about 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, or other ratios within these ranges. The concentration of BCP in the composition can range from about 10, 20, 30, 40, 50, 60, 70, 80, or 90 to about 350, 300, 250, 200, 150, or 100 mg / mL of composition. In one embodiment, the composition contains about 100 mg of CBD and 100 mg of BCP per mL of composition.

[0064]

[0082] Commercial sources of BCP include products available from Sigma Aldrich, the product specifications of which are contained in Appendix A (CAS No. 87-44-5). Such products contain at least 95% w / w of major and minor amounts of C15H24 terpenes and impurities such as manufacturing by-products up to 5% w / w. Sigma Aldrich's BCP therefore has a "purity" of at least 95%. Other embodiments of the present invention may use other commercial sources of BCP with higher purity, for example, BCP with a purity of at least 96, 97, 98, 99, or 99.5%.

[0065]

[0083] The BCP and its derivatives used in the present compositions may be of synthetic or natural origin.

[0066]

[0084] As used herein, BCP "derivatives" refers to C15H24 minor terpene hydrocarbons, including minor terpenes present in Sigma-Aldrich products.

[0067]

[0085] Pharmaceutically acceptable solvent

[0086] The pharmaceutically acceptable solvent is used to solubilize the active pharmaceutical agent in the composition. In parenteral compositions, these Solvent The solvents should preferably be suitable for parenteral administration by subcutaneous (SC), intramuscular (IM), intraperitoneal (IP) and intravenous routes (IV). For lipophilic drugs such as CBD and BCP, these solvents are hydrophobic or lipophilic.

[0068]

[0087] Examples of hydrophobic or lipophilic solvents include natural and synthetic solvents, including medium chain (C6-C12) triglycerides (MCT), long chain (C14-C20) triglycerides, structured lipids, propylene glycol di(caprylic / capric), vegetable oils, N-methyl-2-pyrrolidone (NMP; Pharmasolve), polyoxyethylene castor oil derivatives, dimethylacetamide (DMA), ethanol, glycerin, PEG 300, PEG 400, polyoxyethylene 20 sorbitan monooleate, propylene glycol (PG), polyglycol mono- and diesters of 12-hydroxystearic acid, solvents sold under the trademark Kolliphor™, and mixtures thereof.

[0069]

[0088] "Medium chain triglyceride" refers to an ester of glycerol having three C6-C12 fatty acid chains, where the three fatty acid chains can be the same or different. Medium chain triglycerides are represented by the following formula: [ka]

[0070]

[0089] wherein each x is independently 4, 6, 8, or 10. When x is 4, the chain is referred to as a C6 fatty acid. When x is 6, the chain is referred to as a C8 fatty acid. When x is 8, the chain is referred to as a C10 fatty acid. When x is 10, the chain is referred to as a C12 fatty acid. In various embodiments, within one molecule of triglyceride, each x is the same integer; two x are the same integer and one x is a different integer; or each x is a different integer.

[0071]

[0090] Preferably, the composition contains medium chain triglycerides (MCTs), more preferably those in which C8 and C10 fatty acids make up at least 95% of the composition. The medium chain triglycerides can be synthetic or natural (e.g., produced from fractionated oils such as coconut oil and / or palm kernel oil).

[0072]

[0091] In various embodiments, the medium-chain triglycerides comprise esters of (i) three C8 fatty acids; (ii) three C10 fatty acids; (iii) two C8 fatty acids and one C10 fatty acid; (iv) two C10 fatty acids and one C8 fatty acid; (v) two C8 fatty acids and one C6 fatty acid; (vi) two C10 fatty acids and one C6 fatty acid; (vii) one C8 fatty acid, one C10 fatty acid and one C6 fatty acid; or (viii) any other combination of C6, C8, C10, and C12 fatty acids. In one embodiment, the medium-chain triglycerides comprise two C8 fatty acids and one C10 fatty acid. In one embodiment, the medium-chain triglycerides comprise two C10 fatty acids and one C8 fatty acid.

[0073]

[0092] Those skilled in the art will recognize that mixtures of medium-chain triglycerides may result from any process used to prepare medium-chain triglycerides (e.g., fractionation, hydrogenation). For example, substantially all medium-chain triglycerides obtained from fractionated coconut oil may contain C8 and / or C10 fatty acids. However, some medium-chain triglycerides containing C6 and / or C12 fatty acids may be present.

[0074]

[0093] In one embodiment, the medium chain triglycerides comprise: (i) 0-2% w / w of C6 fatty acids, 65-80% w / w of C8 fatty acids, 20-35% w / w of C10 fatty acids, and 0-2% w / w of C12 fatty acids; (ii) 0-2% w / w of C6 fatty acids, 50-65% w / w of C8 fatty acids, 30-45% w / w of C10 fatty acids, and 0-2% w / w of C12 fatty acids; (iii) 0-2% w / w or (iv) 0-2% w / w C6 fatty acids, 45-55% w / w C8 fatty acids, 30-40% w / w C10 fatty acids, 0-3% w / w C12 fatty acids, and 10-20% w / w succinic acid esters. In one embodiment, the medium chain triglycerides comprise 0-2% w / w C6 fatty acids, 50-65% w / w C8 fatty acids, 30-45% w / w C10 fatty acids, and 0-2% w / w C12 fatty acids, and are commercially available as MIGLYOL® 812 (IOI Oleo GmbH, Herrengraben 31, 20459 Hamburg, Germany). The weight percentages are based on the total fatty acid content of the triglycerides. In one embodiment, the medium chain triglycerides may contain up to 2% w / w C14 fatty acids.

[0075]

[0094] The carrier may comprise one, two, three, four, or more different medium-chain triglycerides. In one embodiment, the carrier comprises a medium-chain triglyceride comprising an ester of two C8 fatty acids and one C10 fatty acid. In one embodiment, the carrier comprises a medium-chain triglyceride comprising an ester of one C8 fatty acid and two C10 fatty acids. In one embodiment, the carrier comprises two different medium-chain triglycerides, a first medium-chain triglyceride comprising an ester of two C8 fatty acids and one C10 fatty acid, and a second medium-chain triglyceride comprising an ester of one C8 fatty acid and two C10 fatty acids. In one embodiment, the carrier comprises a medium-chain triglyceride comprising 0-2% w / w C6 fatty acid, 50-65% w / w C8 fatty acid, 30-45% w / w C10 fatty acid, and 0-2% w / w C12 fatty acid, based on the total fatty acid content of the medium-chain triglyceride.

[0076]

[0095] Triglycerides can be prepared by methods known in the art and are commercially available as MIGLYOL® 810, 812, 818, 829 (IOI Oleo GmbH, Herrengraben 31, 20459 Hamburg, Germany), NEOBEE® 1053, 895, M-5 (Stepan Company, Northfield, IL), and Labrafac (Gattefosse). Vigon International, Inc. also sells MCTs containing a mixture of C8 and C10 triglycerides in a ratio of about 55:45 to about 65:35 (C8:C10).

[0077]

[0096] In another embodiment, the pharmaceutically acceptable solvent is a propylene glycol diester of saturated vegetable fatty acids having chain lengths of C8 and C10 (caprylic and capric acids). An example of such a commercially available carrier is MIGLYOL® 840 (IOI Oleo GmbH, Herrengraben 31, 20459 Hamburg, Germany).

[0078]

[0097] Other pharmaceutically acceptable solvents include, but are not limited to, those marketed under the trademarks Solutol HS 15, Kolliphors™ (formerly Cremophors™), Labrasol, Labrafil, and Gelucire.

[0079]

[0098] Solutol HS 15 is composed of a mixture of lipophilic and hydrophilic compounds (approximately 70% lipophilic consists of polyglycol mono- and diesters of 12-hydroxystearic acid, and approximately 30% hydrophilic consists of polyethylene glycol). Solutol HS 15 is synthesized by reacting 12-hydroxystearic acid with 15 moles of ethylene oxide.

[0080]

[0099] Kolliphors™ (formerly Cremophors™) can also be used in solvents. These are complex mixtures of various hydrophobic and hydrophilic components. Kolliphor™ EL is obtained by reacting 35 moles of ethylene oxide with 1 mole of castor oil and contains approximately 83% hydrophobic components, the main component being glycerol polyethylene glycol ricinoleate. Kolliphor™ RH 40 is obtained by reacting 40 moles of ethylene oxide with 1 mole of hydrogenated castor oil and contains approximately 75% hydrophobic components, the main component being glycerol polyethylene glycol 12-hydroxystearate.

[0081] [000100] Labrasol is a mixture of mono-, di-, and triglycerides and mono- and di-fatty acid esters of PEG 400. Labrasol is synthesized by an alcoholysis / esterification reaction using medium chain triglycerides derived from coconut oil and PEG 400, with the predominant fatty acids being caprylic / capric acid.

[0082] [000101] Labrafil M-1944 CS is a mixture of mono-, di-, and triglycerides and mono- and di-fatty acid esters of PEG 300. Labrafil M-1944 CS is synthesized by alcoholysis / esterification using apricot kernel oil and PEG 300, and the predominant fatty acid is oleic acid (58-80%).

[0083] [000102] Labrafil M-2125 CS is a mixture of mono-, di-, and triglycerides and mono- and di-fatty acid esters of PEG 300. Labrafil M-2125 CS is synthesized by alcoholysis / esterification using corn oil and PEG 300, and the predominant fatty acid is linoleic acid (50-65%).

[0084] [000103] Gelucire 44 / 14 is a mixture of mono-, di-, and triglycerides and mono- and di-fatty acid esters of PEG 1500. Gelucire 44 / 14 is synthesized by an alcoholysis / esterification reaction using palm kernel oil and PEG 1500, and the predominant fatty acid is lauric acid.

[0085] [000104] One of ordinary skill in the art would know how to formulate compositions for SC, IM, and IV administration based on published literature such as Strickley, "Solubilizing Excipients in Oral and Injectable Formulations" (Pharmaceutical Research, Vol. 21, No. 2, February 2004 (© 2004)), which is incorporated herein by reference in its entirety. Components such as preservatives, emulsifiers, tonicity agents, salts, buffers, diluents, and the like may also be used.

[0086] [000105] In a preferred embodiment, the composition comprises an effective amount of CBD or an effective amount of each of CBD and BCP dissolved in MCT, and is substantially free of water and micelles.

[0087] [000106] Method of administration [000107] In some embodiments, The compositions are for parenteral administration, for example, via intravenous (IV), subcutaneous (SC), intraperitoneal (IP), and intramuscular (IM) injection. The following examples demonstrate that the compositions may be useful in reducing leukocyte-mediated inflammation, cardiac fibrosis, and / or cardiac hypertrophy.

[0088] [000108] The composition can be administered at least once a week or at least once every 6, 5, 4, 3, or 2 days. The composition can also be administered at least once a day or at least twice a day. Each administration provides at least about 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 mg of pharmaceutically active agent per kg of body weight. The duration of treatment can be 6, 5, 4, 3, or 2 months or 8, 7, 6, 5, or 4 weeks. For chronic conditions (e.g., heart failure), it is contemplated that very low doses can be administered indefinitely, with the dosage selected to avoid long-term toxicity. In one embodiment, 1 mg / kg of body weight can be administered at least once a week or at least once every 1, 2, 3, 4, or 5 days indefinitely. [Example]

[0089] [000109] Working Example [000110] Example 1 - CBD reduces angiotensin II-mediated hypertrophy in H9c2 cells in vitro [000111] An in vitro study was conducted to measure the effects of CBD in a model of cardiomyocyte hypertrophy using H9c2 cells, a cardiomyocyte cell line. In this study, H9c2 cells grown in culture medium were divided into four groups. Group 1 received nothing ("Control"). Groups 2, 3, and 4 received angiotensin II ("Ang") in the culture medium to a final concentration of 1.0 μM. Angiotensin II was added to induce hypertrophy. Group 3 also received CBD to a concentration of 0.1 μM ("Ang+CBD 0.1 μM"). Finally, group 4 also received CBD to a concentration of 1 μM ("Ang+CBD 1 μM").

[0090] [000112] Figure 1 includes representative confocal microscopy images of H9c2 cells from each group. Images were obtained using a model with the same parameters, and cell staining to measure surface area per cell was achieved using calcein fluorescent dye. As can be seen in this figure, cells from group 2 had a significantly larger cell surface area than cells from group 1. Cells from groups 3 and 4 had significantly smaller cell surface areas than those from group 2. The results are summarized in Figure 2, and each group contains at least three independent replicates per group.

[0091] [000113] Thus, the study demonstrated that CBD exerts an effect on cardiac cells and is effective in reducing the degree of hypertrophy caused by angiotensin II administration to H9c2 cells in vitro.

[0092] [000114] The study was repeated using H9c2 cells grown in culture medium divided into seven groups. To one group, nothing was added ("CTRL"). To groups 2, 4, 5, 6, and 7, angiotensin II was added to the culture medium ("ANG") to a final concentration of 1.0 μM. Angiotensin II was added to induce hypertrophy. To group 3, CBD was added to a concentration of 0.1 μM ("CBD 0.1 μM") as an additional control group. CBD was also added to (a) group 4 at a concentration of 0.001 μM ("ANG+CBD 0.001 μM"), (b) group 5 at a concentration of 0.01 μM ("ANG+CBD 0.01 μM"), (c) group 6 at a concentration of 0.1 μM ("ANG+CBD 0.1 μM"), and finally (d) group 7 at a concentration of 1 μM ("ANG+CBD 1 μM").

[0093] [000115] In these H9c2 cell groups, there was a dose-dependent reduction in the mRNA levels of BNP (a cardiac remodeling biomarker) (see Figure 3a) and collagen (see Figure 3b). BNP was significantly reduced in the ANG + CBD 1 μM group compared to the ANG group (Figure 3a), and collagen was significantly reduced in the angiotensin-treated CBD groups at 0.1 and 1 μM concentrations compared to the ANG group (Figure 3b). Mitochondrial reactive oxygen species (ROS), a marker of inflammation, were also reduced to control levels (see Figure 4).

[0094] [000116] These experiments demonstrate that CBD exerts an antihypertrophic effect on cardiac cells, reflected at the molecular level by a significant reduction in mitochondrial ROS, a biomarker of remodeling and a mediator of inflammation.

[0095] [000117] Example 2 - CBD reduces fibrosis, myocyte area, remodeling parameters, and pro-inflammatory cytokines and increases anti-inflammatory cytokines in a mouse model of cardiac inflammation and heart failure [000118] A study was conducted to evaluate the ability of CBD to treat or prevent heart failure in a mouse model of non-ischemic heart failure (HF) depicted in Figure 5.

[0096] [000119] Male C57BL / 6 laboratory mice, 3-4 weeks old, were used in this experiment. The mice were divided into four groups. Group 1 (control) received no supplements. Group 2 (angiotensin) mice received water containing 0.1 mg / ml N(γ)-nitro-L-arginine methyl ester (L_NAME) and 1% sodium chloride (NaCl) ad libitum for one week. This was followed by subcutaneous administration of 0.7 mg / kg / week of angiotensin II using an osmotic pump for four weeks. Groups 3 and 4 underwent the same protocol as Group 2, except that Group 3 (ANG + CBD 1 mg / kg) and Group 4 (ANG + CBD 10 mg / kg) received synthetic CBD subcutaneously in amounts of 1 mg / kg and 10 mg / kg, respectively, once every three days for the same four-week period. The CBD composition used in this experiment consisted of CBD dissolved in PEG 400.

[0097] [000120] After 4 weeks of treatment, cardiac tissue was evaluated for fibrosis and myocyte hypertrophy as well as changes in remodeling and inflammatory biomarkers.

[0098] [000121] Several changes, including increased fibrosis and increased myocyte area (hypertrophy), were observed in the hearts of mice in Groups 2, 3, and 4 compared to Group 1. Figures 6A, 6B, 6C, and 6D show representative slides of cardiac tissue from each of the four groups. Cardiac tissue was stained with Masson's Trichrome stain to demonstrate increased levels of fibrotic tissue (evidenced by blue staining) in Groups 2, 3, and 4 compared to Group 1 (control), and reduced levels of fibrosis in Groups 3 and 4 compared to Group 2. This information is graphically represented in Figure 7. Myocyte area was also measured in all four groups, and the results are also shown in Figure 8. Figures 7 and 8 show that both 1 mg / kg and 10 mg / kg CBD were effective in reducing cardiac fibrosis and myocyte area in a mouse model of heart failure. Each group contained at least three independent replicates.

[0099] [000122] Figure 9 shows the mRNA expression levels of B-type natriuretic peptide (BNP) in cardiac tissue of mice from all four groups, measured by qPCR. Both the 1 mg / kg and 10 mg / kg CBD groups showed significantly reduced BNP expression in the heart compared to the ANG group. Each group contained at least three independent replicates. BNP is a clinical marker of remodeling and myocardial stretch (pressure overload), and it is positively correlated with cardiac dysfunction and heart failure (see, e.g., Glezeva et al.). There is clinical evidence that reducing BNP levels to less than 100 pg / mL in patients with chronic heart failure (CHF) reduces the risk of CHF-related death or hospitalization (Jourdain et al., 2007: https: / / www.ncbi.nlm.nih.gov / pubmed / ?term=17448376). Therefore, this reduction in BNP may also indicate that CBD may be beneficial in the treatment and / or prevention of heart failure.

[0100] [000123] Furthermore, mRNA expression of the inflammatory cytokine IL1β tended to decrease to the control level (Fig. 10a), and mRNA expression of the inflammatory cytokine IL6 was significantly decreased by both 1 and 10 mg / kg body weight doses of CBD (Fig. 10b). mRNA expression of IL10 was significantly increased by 1 mg / kg body weight dose of CBD (Fig. 10c). Each group contained at least three independent replicates. IL10 is an anti-inflammatory cytokine and is also known as human cytokine inhibitory factor, which can inhibit the synthesis of inflammatory cytokines. Evidence suggests that IL-10 plays a role in vascular protection; in animal models, a lack of IL-10 has been shown to cause COX-2 / thromboxane A2-dependent endothelial and cardiac dysfunction (Gautam Sikka et al., 'Interleukin 10 Knockout Frail Mice Develop Cardiac and Vascular Dysfunction with Increased Age', Experimental Gerontology 48, no. 2 (February 2013): 128, https: / / doi.org / 10.1016 / j.exger.2012.11.001).

[0101] [000124] In summary, the present inventors observed a dose-dependent reduction in remodeling and inflammatory parameters resulting from the administration of CBD in a mouse model of non-ischemic HF. Regarding remodeling, significant reductions in fibrosis, myocyte hypertrophy, and BNP gene expression were observed. Inflammatory parameters improved, i.e., CBD administration reduced the levels of inflammatory cytokines IL1 and IL6 and significantly increased the level of the anti-inflammatory cytokine IL10. These findings support the beneficial role of CBD in the treatment or prevention of heart failure and other pathologies, such as acute myocarditis, inflammatory cardiomyopathy, cardiac sarcoidosis, acute pericarditis, myocardial damage resulting from the administration of certain anticancer drugs, and possibly the development or progression of coronary atherosclerotic lesions, by reducing cardiac inflammation and associated hypertrophy and fibrosis.

[0102] [000125] Example 3 - CBD reduces activation of human lymphocytes and monocytes [000126] Further studies were performed to measure the effect of CBD on the expression of CD69 (a marker of leukocyte activation) by human lymphocytes and monocytes activated in vitro with ionomycin (1 μg / mL) and phorbol myristate acetate (PMA, 50 ng / mL).

[0103] [000127] Mononuclear cells from healthy human donors were isolated using a gradient method with Ficoll-Pacque HE Healthcare. Cells were washed with RPMI medium containing 10% FCS and 1% penicillin / streptomycin, and then plated at 1 x 10 cells per well in a 12-well tray. 6 Cells were plated at 1000 cells / mL and stimulated with 1 μg / mL ionomycin and 50 ng / mL PMA for 4 hours. CBD was added 15 minutes before DMSO was used as a vehicle. After stimulation, cells were washed again and resuspended in 150 μL staining buffer (PBS + 1% BSA). An optimized protocol for surface marker staining was used. CD69 was added to the cells, which were then incubated for 15 minutes in the dark at room temperature. After this, cells were washed twice with PBS + 1% BSA and resuspended in 150 μL buffer. Cells were kept at 4°C before analysis on a FACSCanto™ II cytometer. Cell populations were measured by FSC / SSC. The number of events was at least 2000 events per gate. Data were saved as FCS3.0 files. Data were analyzed using FlowJo X. Doublets were distinguished using an FSC-A / FSC-W exclusion gate. Gating of lymphocytes and monocytes was determined by FSC-A / SSC-A. The frequency and expression of CD69-FITC were analyzed in the FITC channel.

[0104] [000128] Figure 11 shows that administration of CBD at 5 μg / mL or more (per mL of cell culture medium containing RPMI with 10% fetal bovine serum and 1% penicillin / streptomycin) reduced the level of CD69 expression by both monocytes and lymphocytes. Thus, this experiment indicates that CBD can reduce leukocyte activation in vitro and is expected to reduce inflammation, one of the underlying pathologies of heart failure.

[0105] [000129] INDUSTRIAL APPLICABILITY [000130] The experiments described herein demonstrate that CBD is useful in the treatment or prevention of cardiac conditions selected from the group consisting of heart failure (e.g., HFpEF), acute myocarditis, toxicity caused by anti-cancer therapy (e.g., doxorubin, checkpoint inhibitors), acute pericarditis, cardiac sarcoidosis, inflammatory cardiomyopathy, and atherosclerosis by reducing cardiac inflammation and associated hypertrophy and fibrosis.

[0106] [000131] Acute inflammation is a defensive response of the body's immune system to danger signals, including infected and damaged cells, to eliminate infection and repair damaged tissue. Inflammation leads to increased blood flow and vascular permeability, allowing immune cells to access the site of infection or injury. After tissue repair and removal of danger signals, it is essential to switch off the initial inflammatory and repair responses. This is usually the result of accumulated inflammatory cells undergoing cell death by apoptosis and their engulfment by macrophages by efferocytosis, a process that triggers an anti-inflammatory response and stops inflammation. Failure to switch off this switch leads to continued chronic inflammation, which is seen in many diseases, including heart failure.

[0107] [000132] As mentioned above, factors that predispose people to heart failure include aging, diabetes, and obesity. These conditions are associated with increased low-grade background inflammation. Without being bound by theory, it is believed that chronic inflammation results, inter alia, from impaired efferocytosis of apoptotic cells, leading to increased cardiomyocyte (heart muscle cell) cell death, increased fibrosis (accumulation of scar tissue), and decreased cardiac function due to the resulting weakening and stiffening of the myocardium.

[0108] [000133] Acute myocarditis is the leading cause of sudden cardiac death in people under 35. It is characterized by inflammation within the heart muscle (myocardium). It has many causes, the most common being a viral infection. In most patients, the immune system is effective in eliminating the virus within 5 to 7 days, the inflammation subsides, and the individual recovers fully. However, in some patients, the inflammation within the heart—possibly as an autoimmune process—continues, causing a decline in cardiac function along with symptoms and signs of heart failure. In some cases, this becomes progressive, leading to chronic dilated cardiomyopathy, the most common reason for heart transplantation.

[0109] [000134] The above-described embodiments are merely illustrative and do not limit the scope of the invention as described in detail herein and defined by the following claims.

[0110] [Table 1]

Claims

1. 1. A pharmaceutical composition for use in the treatment of heart failure, comprising: The pharmaceutical composition is a parenteral composition comprising an effective amount of cannabidiol and an effective amount of at least one solvent to solubilize the cannabidiol in the pharmaceutical composition, wherein the pharmaceutical composition is substantially water-free and micelle-free.

2. The pharmaceutical composition of claim 1, wherein the at least one solvent is selected from the group consisting of natural and synthetic medium-chain (C6 to C12) triglycerides (MCTs).

3. 3. The pharmaceutical composition of claim 2, wherein at least about 95% of the MCTs consist of C8 triglycerides, C10 triglycerides, or a mixture thereof.

4. The pharmaceutical composition of any one of claims 1 to 3, wherein the composition further comprises an effective amount of at least one additional pharmaceutically active agent.

5. 5. The pharmaceutical composition of claim 4, wherein the at least one additional pharmaceutically active agent is β-caryophyllene.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the parenteral composition is adapted for subcutaneous or intramuscular administration.

7. 7. The pharmaceutical composition of any one of claims 1 to 6, wherein the cannabidiol is present in an amount of up to about 350 mg / mL of the composition.

8. 8. The pharmaceutical composition of any one of claims 1 to 7, wherein the cannabidiol is present in an amount from about 10 mg / mL of the composition.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the heart failure is heart failure with preserved ejection fraction (HFpEF).

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