Synthetic cannabinoid analogs, pharmaceutical compositions and methods of treating anxiety and other disorders

Abstract

Cannabinoid analogs may exhibit anti-inflammatory properties such as by inhibition of cannabinoid type 2 (CB2) receptors. Pharmaceutical compositions comprising the cannabinoid analogs may be used to treat various diseases and conditions in mammals, including pain, anxiety, a sleep disorder, addiction, epilepsy, depression, post-traumatic stress disorder or Alzheimer's disease. In some examples, the pharmaceutical compositions may be administered for treating a cognitive disorder or for improving cognition.

Description

[0001] Atty DktNo. 009806.00123

[0002] SYNTHETIC CANNABINOID ANALOGS, PHARMACEUTICAL COMPOSITIONS AND METHODS OF TREATING ANXIETY AND OTHER DISORDERS

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004]

[0001] This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional App. No. 63 / 557,913, filed February 26, 2024, and U.S. Provisional App. No. 63 / 557,934, filed February 26, 2024, the disclosure of each of which is hereby incorporated by reference in its entirety.

[0005] BACKGROUND

[0006]

[0002] There has been considerable research in recent years on the therapeutic effects of cannabis including its constituents tetrahydrocannabinol (THC) and cannabidiol (CBD). There remains a need for improved compounds for treating disorders such as depression, anxiety, substance addiction, sleep disorder, pain, cancer, autoimmune disorders, and other disorders associated with chronic inflammation. It would be particularly desirable to develop compounds which may be prepared synthetically and formulated as solid oral dosage forms.

[0007] SUMMARY

[0008]

[0003] According to one aspect, a synthetic cannabinoid analog has a structure of Formula (I): wherein Ri and R2 are each independently selected from the group consisting of H, OH, protected hydroxyl, alkyl, alkenyl, alkynyl, acyl, aryl, heteroaryl, cycloalkyl, and heterocycle; wherein the alkyl, alkenyl, alkynyl or acyl is optionally substituted with Atty DktNo. 009806.00123 one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRARB, — S-alkyl, — SO-alkyl, — SO2-alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, and heterocycle; wherein RAand RBare each independently selected from hydrogen and Ci-4 alkyl; wherein the aryl or heteroaryl, whether alone or as part of a substituent group, is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, — COOH, — C(O)— Ci-4 alkyl, — C(O)O— Ci-4 alkyl, NRCRD, —S-alkyl, —SO-alkyl and — SO2-alkyl; wherein Rcand RDare each independently selected from hydrogen and C1-4 alkyl;

[0009] R3 is selected from the group consisting of H, alkyl, acyl, — SO2-alkyl, — SO2-aryl and — SO2- heteroaryl; wherein the alkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRERF, — S-alkyl, — SO-alkyl, — SO2-alkyl, aryl and heteroaryl; and wherein REand REare each independently selected from hydrogen and C1-4 alkyl; wherein the aryl or heteroaryl, whether alone or as part of a substituent group, is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRGRH, — S-alkyl, — SO-alkyl and — SO2-alkyl; wherein RGand RHare each independently selected from hydrogen and C1-4 alkyl; each -> represents a single or double bond, with the proviso that within a 5- membered ring, one or two - is a double bond and the remaining > - are single bonds; or a pharmaceutically acceptable salt or ester thereof.

[0010]

[0004] According to another aspect, a method of treating a cancer, tumor, Alzheimer’s disease, addiction, epilepsy, anxiety, sleep disorder, pain, post-traumatic stress disorder (PTSD) or depression comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a synthetic cannabinoid analog of Formula (I) and a pharmaceutically acceptable carrier therefor.

[0011]

[0005] In some aspects, a synthetic cannabinoid analog has a structure of Formula (IA): Atty DktNo. 009806.00123 wherein Ri, R2, and R3 are as previously defined; and one is a double bond and the other is a single bond.

[0012]

[0006] According to another aspect, a method of treating a cancer, tumor, addiction, epilepsy, anxiety, sleep disorder, PTSD or depression comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a synthetic cannabinoid analog of Formula (IA) and a pharmaceutically acceptable carrier therefor.

[0013]

[0007] According to other aspects, a synthetic cannabinoid analog has a structure of Formula (II):

[0014] Formula (II) wherein R2 is as previously defined, or a pharmaceutically acceptable salt or ester thereof. In some examples, R2 is C1-C4 straight-chained or branched alkyl, e.g., propyl. In other examples, R2 is C5-C10 straight-chained or branched alkyl, e.g., pentyl or heptyl. Atty DktNo. 009806.00123

[0015]

[0008] In another aspect, a method of treating anxiety, addiction, depression, sleep disorder, or PTSD comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a synthetic cannabinoid analog of Formula (II) and a pharmaceutically acceptable carrier therefor.

[0016]

[0009] In another aspect, a pharmaceutical composition comprises a therapeutically effective amount of a synthetic cannabinoid analog of Formula (I), (IA), or (II), or a combination thereof, and a pharmaceutically acceptable carrier therefor.

[0017]

[0010] In some embodiments, the compound disclosed herein has the structure: Atty DktNo. 009806.00123

[0018] , or a pharmaceutically acceptable salt or ester thereof.

[0019]

[0011] In one aspect, a pharmaceutical composition comprises a therapeutically effective amount of a compound having the structure: or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable vehicle therefor. In some examples the pharmaceutical composition is administered to an individual in need thereof for the treatment of a sleep disorder. In other examples, the pharmaceutical composition is administered to an individual in need thereof for the treatment of an addiction, such as smoking or smokeless tobacco addiction.

[0020]

[0012] In another aspect, a pharmaceutical composition comprises a therapeutically effective amount of a compound having the structure: or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable vehicle therefor. In some examples the pharmaceutical composition is administered to an individual in need thereof for the treatment of a cancer, tumor, Alzheimer’s disease, addiction, epilepsy, anxiety, sleep disorder, pain, post-traumatic stress disorder (PTSD) or depression. In other examples, the pharmaceutical Atty DktNo. 009806.00123 composition is administered to an individual in need thereof to treat a cognitive disorder or to improve cognition.

[0021]

[0013] In yet another aspect, a pharmaceutical composition comprises a therapeutically effective amount of a compound having the structure: or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable vehicle therefor. In some examples the pharmaceutical composition is administered to an individual in need thereof for the treatment of PTSD.

[0022] BRIEF DESCRIPTION OF DRAWINGS

[0023]

[0014] FIG. 1 shows the response curves of increasing concentration of test compound 3a- isopropyl-2-methyl-6-propyl-3a,8b-dihydro-lH-cyclopenta[b]benzofuran-8-ol (T-55, circular symbols) and reference compound R(-)-Deprenyl (square symbols) as indicated on the x-axis. Percentage inhibition of MAO-B is shown on the y-axis.

[0024]

[0015] FIG. 2 shows representative monitoring of intermediate 5 cyclization to the target compound M-55 as described in Example 3, trial 7.

[0025]

[0016] FIG. 3 shows the HPLC chromatogram for M-55 as synthesized in Example 3, trial 13.

[0026]

[0017] FIG. 4 shows the MS spectrum for M-55 as synthesized in Example 3, trial 13.

[0027]

[0018] FIG. 5 shows the 'H-NMR results for M-55 as synthesized in Example 3, trial 13.

[0028]

[0019] FIG. 6 shows the13C-NMR results for M-55 as synthesized in Example 3, trial 13.

[0029]

[0020] FIG. 7 shows the HPLC chromatogram for M-55 as synthesized in Example 3, trial 16.

[0030]

[0021] FIG. 8 shows the MS spectrum for M-55 as synthesized in Example 3, trial 16. Atty DktNo. 009806.00123

[0031]

[0022] FIG. 9 shows the1H-NMR results for M-55 as synthesized in Example 3, trial 16.

[0032]

[0023] FIG. 10 shows the13C-NMR results for M-55 as synthesized in Example 3, trial 16.

[0033]

[0024] FIG. 11 shows group means (+SEM) effects of M-55 (3, 10, 30, 100 mg / kg, p.o.) or vehicle (sesame oil, p.o.) on inflammation-induced thermal pain (a), mechanical pain

[0034] (b), and paw edema (c); N= 8-10 per sex / dose.

[0035]

[0025] FIG. 12 shows group means (+SEM) effects of M-55 or vehicle on thermal hyperalgesia

[0036] 1-hr after carrageenan; N=16-20 per group; sexes combined. P-values shown were obtained using the Dunnett’s post-hoc test in comparison with vehicle.

[0037]

[0026] FIG. 13 shows group means (+SEM) effects of THC (1, 3, and 10 mg / kg, p.o.) or vehicle

[0038] (sesame oil p.o.) on inflammation-induced thermal pain (a), mechanical pain (b), and paw edema (c); N= 8-10 per sex / dose.

[0039]

[0027] FIG. 14 shows group means (+SEM) effects of THC (1, 3, and 10 mg / kg, p.o.) or vehicle

[0040] (Sesame oil, p.o.) on thermal hyperalgesia 1-hr after carrageenan. N=16-20 per group; sexes combined.

[0041]

[0028] FIG. 15 shows group means (+SEM) effects of Ketoprofen (10, 20 mg / kg, i.p.) or vehicle

[0042] (i.p.) on inflammation-induced thermal pain (a), mechanical pain (b), and paw edema

[0043] (c); N=8-12 per sex / dose.

[0044]

[0029] FIG. 16 shows group means (+SEM) effects of Ketoprofen or vehicle on thermal hyperalgesia 5-hrs after carrageenan; N=19-23 per group; sexes combined.

[0045]

[0030] FIG. 17 shows group means (+SEM) effects of M-55 (3, 10, 30 and 100 mg / kg, p.o.) or vehicle (sesame oil, p.o.) on acute thermal pain sensitivity (a) and mechanical pain sensitivity (b); N= 6 per sex / dose.

[0046]

[0031] FIG. 18 shows peak antinociceptive effects of M-55 (3, 10, 30 and 100 mg / kg, p.o.) vs. vehicle (sesame oil, p.o.) in tests of thermal pain sensitivity (a) and mechanical pain sensitivity (b), 60 and 30-minutes after drug administration, respectively; N=6 per sex / dose. Data are group means (+SEM).

[0047]

[0032] FIG. 19 shows the results of M-55 in an Elevated Plus Maze (EPM) anxiety test. Atty DktNo. 009806.00123

[0048] DETAILED DESCRIPTION

[0049]

[0033] Cannabinoids produced by the Cannabis sativa plant have the potential to treat a vast assortment of diseases and other human ailments. More than 100 different cannabinoids have been isolated from cannabis and each cannabinoid compound exhibits various effects. For example, THC is well-known for its psychological effects and CBD is known for its non-psychoactive effects. THC and related analogs typically exert therapeutic activities via cannabinoid receptors found in humans and other mammals. CBD is an isomer of THC. CBD and CBD derivatives also exhibit anti-oxidative and anti-inflammatory effects through pathways not related to cannabinoid receptors. Cannabinoid type 1 (CBi) receptors are found primarily in the brain, including the basal ganglia and in the limbic system, and the hippocampus and the striatum, as well as the cerebellum. CBi receptors can be found in the human anterior eye and retina. Research indicates that cannabinoid type 2 (CB2) receptors are responsible for anti-inflammatory and other therapeutic effects related to cannabinoids.

[0050]

[0034] Cannabis plants that contain high levels of cannabinoids such as THC, for example, are typically known as “marijuana” plants. Cannabis plants with a low cannabinoid content are categorized as “hemp” plants. Individual countries usually determine the levels of cannabinoids that differentiate between cannabis plants that are categorized as marijuana or hemp plants. Generally, the THC content on a dry -weight basis for a cannabis plant categorized as a hemp plant is 0.3% or less. Cannabis sativa plants having THC, CBD, and other cannabinoid content levels greater than 0.3% are typically considered marijuana plants. Medical marijuana typically contains cannabinoid levels between 5 and 20%. Other Cannabis sativa plants may produce cannabinoid levels from 25 to 30%.

[0051]

[0035] The biosynthetic pathway of the Cannabis sativa plant that produces the various cannabinoids starts with the precursor cannabigerolic acid. The enzymes THCA synthase and CBDA synthase catalyze the biosynthesis of cannabigerolic acid to tetrahydrocannabinol acid (THCA) and cannabidiol acid (CBDA), respectively, as well as other cannabinoids. It is known that various other cannabinoids are produced via this pathway. THC, CBD, and other cannabinoid derivatives are generated artificially from THCA and CBDA by non-enzymatic decarboxylation. Aizpurua-Olaizola et al., “Evolution of the Cannabinoid and Terpene Content during the Growth of Cannabis Atty DktNo. 009806.00123 sativa Plants from Different Chemotypes,” J. Natural Prods. 2016 79 (2), 324-331. Various classes of cannabinoids are biosynthesized via this general pathway to include THC (tetrahydrocannabinol), THCA (tetrahydrocannabinolic acid), CBD (cannabidiol), CBDA (cannabidiolic Acid), CBN (cannabinol), CBG (cannabigerol), CBC (cannabichromene), CBL (cannabicyclol), CBV (cannabivarin), THCV (tetrahydrocannabivarin), CBDV (cannabidivarin), CBCV (cannabichromevarin), CBGV (cannabigerovarin), CBGM (cannabigerol monomethyl ether), CBE (cannabielsoin), and CBT (cannabicitran).

[0052] I. Synthetic cannabinoid analogs

[0053]

[0036] According to some aspects, a synthetic cannabinoid analog has a structure of Formula

[0054] (I): wherein Ri and R2 are each independently selected from the group consisting of H, OH, protected hydroxyl, alkyl, alkenyl, alkynyl, acyl, aryl, heteroaryl, cycloalkyl, and heterocycle; wherein the alkyl, alkenyl, alkynyl or acyl is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRARB, — S-alkyl, — SO-alkyl, — SO2-alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, and heterocycle; wherein RAand RBare each independently selected from hydrogen and C1-4 alkyl; wherein the aryl or heteroaryl, whether alone or as part of a substituent group, is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, — COOH, — C(O)— Ci-4 alkyl, — C(O)O— Ci-4 alkyl, NRCRD, —S-alkyl, —SO-alkyl Atty DktNo. 009806.00123 and — SCh-alkyl; wherein Rcand RDare each independently selected from hydrogen and Ci-4 alkyl;

[0055] R3 is selected from the group consisting of H, alkyl, acyl, — SCh-alkyl, — SCh-aryl and — SO2- heteroaryl; wherein the alkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRERF, — S-alkyl, — SO-alkyl, — SO2-alkyl, aryl and heteroaryl; and wherein REand REare each independently selected from hydrogen and C1-4 alkyl; wherein the aryl or heteroaryl, whether alone or as part of a substituent group, is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRGRH, — S-alkyl, — SO-alkyl and — SO2-alkyl; wherein RGand RHare each independently selected from hydrogen and C1-4 alkyl; each - represents a single or double bond, with the proviso that within a 5 -membered ring, one or two - is a double bond and the remaining ->> are single bonds; or a pharmaceutically acceptable salt or ester thereof.

[0056]

[0037] In other examples, a synthetic cannabinoid analog has a structure of Formula (IA):

[0057] Formula (I A) wherein Ri, R2, R3 are as previously defined; and one - is a single bond and the other

[0058] - is a double bond; or a pharmaceutically acceptable salt or ester thereof. Atty DktNo. 009806.00123

[0059]

[0038] According to other aspects, a synthetic cannabinoid analog has a structure of Formula (II):

[0060] Formula (II) wherein R2 is as previously defined, or a pharmaceutically acceptable salt or ester thereof. In some examples, R2 is C1-C4 straight-chained or branched alkyl, e.g., propyl. In some examples, R2 is pentyl. In other examples, R2 is Ce-Cio straight-chained or branched alkyl, e.g., hexyl or heptyl.

[0061]

[0039] As used herein the term “alkyl,” whether alone or as part of a substituent group, refers to a saturated Ci-Cn carbon chain, wherein the carbon chain may be straight or branched; wherein n can be 2, 3, 4, 5, 6, 7, 8, 9 or 10. Suitable examples include, but are not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, n-pentyl and n- hexyl.

[0062]

[0040] As used herein the term “alkenyl,” whether alone or as part of a substituent group, refers to a C2-C11 carbon chain, wherein the carbon chain may be straight or branched, wherein the carbon chain contains at least one carbon-carbon double bond, and wherein n can be 3, 4, 5, 6, 7, 8, 9 or 10.

[0063]

[0041] As used herein the term “alkynyl,” whether alone or as part of a substituent group, refers to a C2-C11, wherein the carbon chain may be straight or branched, wherein the carbon chain contains at least one carbon-carbon triple bond, and wherein n can be 3, 4, 5, 6, 7, 8, 9 or 10. Atty DktNo. 009806.00123

[0064]

[0042] As used herein the term “aryl,” whether alone or as part of a substituent group, refers to an unsubstituted carboxylic aromatic ring comprising between 6 to 14 carbon atoms. Suitable examples include, but are not limited to, phenyl and naphthyl.

[0065]

[0043] As used herein the term “protected hydroxyl” refers to a hydroxyl group substituted with a suitably selected oxygen protecting group. More particularly, a “protected hydroxyl” refers to a substituent group of the formula — OPGi wherein PGi is a suitably selected oxygen protecting group. During any of the processes for preparation of the compounds of the present disclosure it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. J. F. W. McOmie, Plenum Press, 1973; and T. W. Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991. The protecting groups may be removed at a convenient subsequent stage using methods known from the art.

[0066]

[0044] As used herein the term “oxygen protecting group” refers to a group which may be attached to an oxygen atom to protect said oxygen atom from participating in a reaction and which may be readily removed following the reaction. Suitable oxygen protecting groups include, but are not limited to, acetyl, benzoyl, t-butyl-dimethylsilyl, trimethylsilyl (TMS), MOM and THP. Other suitable oxygen protecting groups may be found in texts such as T. W. Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991.

[0067]

[0045] As used herein the term “nitrogen protecting group” refers to a group which may be attached to a nitrogen atom to protect said nitrogen atom from participating in a reaction and which may be readily removed following the reaction. Suitable nitrogen protecting groups include, but are not limited to, carbamates groups of the formula — C(O) — OR wherein R can be methyl, ethyl, t-butyl, benzyl, phenylethyl, CH2=CH — CH2 — , and the like; amide groups of the formula — C(O) — R' wherein R' can be methyl, phenyl, trifluoromethyl, and the like; N-sulfonyl derivative groups of the formula — SO2 — R" wherein R" can be tolyl, phenyl, trifluoromethyl, 2,2,5,7,8-pentamethylchroman-6-yl-, 2,3,6-trimethyl-4-methoxybenzene, and the like. Other suitable nitrogen protecting Atty DktNo. 009806.00123 groups may be found in texts such as T. W. Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991.

[0068]

[0046] As used herein the term “acyl” refers to a group of the formula — CO — Cn wherein Cn represent a straight or branched alkyl chain wherein n can be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0069]

[0047] As used herein the term “heteroaryl” refers to any five or six membered monocyclic aromatic ring structure containing at least one heteroatom selected from the group consisting of O, N and S, and optionally containing one to three additional heteroatoms independently selected from the group consisting of O, N and S; or a nine or ten membered bicyclic aromatic ring structure containing at least one heteroatom selected from the group consisting of O, N and S, and optionally containing one to four additional heteroatoms independently selected from the group consisting of O, N and S. The heteroaryl group may be attached at any heteroatom or carbon atom of the ring such that the result is a stable structure. Examples of suitable heteroaryl groups include, but are not limited to, pyrrolyl, furyl, thienyl, oxazolyl, imidazolyl, purazolyl, isoxazolyl, isothiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyranyl, furazanyl, indolizinyl, indolyl, isoindolinyl, indazolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthiazolyl, purinyl, quinolizinyl, quinolinyl, isoquinolinyl, isothiazolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl and pteridinyl.

[0070]

[0048] As used herein the term “cycloalkyl” refers to any monocyclic ring containing from four to six carbon atoms, or a bicyclic ring containing from eight to ten carbon atoms. The cycloalkyl group may be attached at any carbon atom of the ring such that the result is a stable structure. Examples of suitable cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0071]

[0049] As used herein the term “heterocycle” refers to any four to six membered monocyclic ring structure containing at least one heteroatom selected from the group consisting of O, N and S, and optionally containing one to three additional heteroatoms independently selected from the group consisting of O, N and S; or an eight to ten membered bicyclic ring structure containing at least one heteroatom selected from the group consisting of O, N and S, and optionally containing one to four additional heteroatoms independently Atty DktNo. 009806.00123 selected from the group consisting of O, N and S. The heterocycle group may be attached at any heteroatom or carbon atom of the ring such that the result is a stable structure. Examples of suitable heterocycle groups include, but are not limited to, azetidine, azete, oxetane, oxete, thietane, thiete, diazetidine, diazete, dioxetane, dioxete, dithietane, dithiete, pyrrolidine, pyrrole, tetrahydrofuran, furan, thiolane, thiophene, piperidine, oxane, thiane, pyridine, pyran and thiopyran.

[0072]

[0050] The groups described herein can be unsubstituted or substituted, as herein defined. In addition, the substituted groups can be substituted with one or more groups such as a Ci- Ce alkyl, Ci-4 alkyl, — O — Ci-4 alkyl, hydroxyl, amino, (Ci-4 alkyl)amino, di(Ci-4 alkyl)amino, — S — (Ci-4 alkyl), — SO — (Ci-4 alkyl), — SO2 — (C1-4 alkyl), halogen, aryl, heteroaryl, and the like.

[0073]

[0051] With reference to substituents, the term “independently” means that when more than one of such substituents is possible, such substituents may be the same or different from each other.

[0074]

[0052] The compounds of the present disclosure may contain at least one hydroxyl group. These at least one hydroxyl group may form an ester with inorganic or organic acid. In particular, pharmaceutically acceptable acids. The ester(s) may form chiral carbons. The present disclosure is directed toward all stereo-chemical forms of the compounds of the present disclosure, including those formed by the formation of one or more ester groups.

[0075] II. Non-limiting examples of synthetic cannabinoid analogs in accordance with the present disclosure

[0076]

[0053] Non-limiting examples of synthetic cannabinoid analogs according to Formulas I, IA and II are illustrated below:

[0077] 3a-isopropyl-2-methyl-6-propyl-3a,8b-dihydro-lH-cyclopenta[b]benzofuran-8-ol Atty DktNo. 009806.00123

[0078] 3a-isopropyl-2-methyl-6-pentyl-3a,8b-dihydro-l / / -cyclopenta[6]benzofuran-8-ol

[0079] 6-heptyl-3a-isopropyl-2-methyl-3a,8b-dihydro- lH-cyclopenta[ / >]benzofuran-8-ol

[0080] III. Synthesis and purification of the cannabinoid compounds

[0081]

[0054] In some examples, the cannabinoid compounds described herein may be formed as salts, which may be helpful to improve chemical purity, stability, solubility, and / or bioavailability. Non-limiting examples of possible salts are described in P. H. Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim / Zurich:Wiley-VCH / VHCA, 2002, including salts of 1 -hydroxy-2 -naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4- acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid (L), aspartic acid (L), benzenesulfonic acid, benzoic acid, camphoric acid (+), camphor- 10- sulfonic acid (+), capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid (D), gluconic acid (D), glucuronic acid (D), glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid (DL), lactobionic acid, lauric acid, maleic acid, malic acid (- L), malonic acid, mandelic acid (DL), Atty DktNo. 009806.00123 methanesulfonic acid, naphthalene-l,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, pyroglutamic acid (- L), salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid (+ L), thiocyanic acid, toluenesulfonic acid (p), and undecylenic acid.

[0082]

[0055] The compounds described herein may be prepared synthetically using known techniques with appropriate modifications to the reactants to form the structures shown herein or by other suitable pathways that will be apparent to persons skilled in the art. By way of nonlimiting example, compounds described herein may be synthesized according to one or more of the following pathways described in Razdan, Total Synthesis of Cannabinoids, SISA Incorporated, Cambridge, Massachusetts, with appropriate modifications to the reactants, as will be apparent to persons skilled in the art, to yield the structures disclosed herein. Alternatively, the synthesis techniques described in Dialer et al. U.S. Patent 10,059,683 B2, the disclosure of which is hereby incorporated by reference in its entirety, may be suitably adapted to synthesize the cannabinoid analogs described herein.

[0083]

[0056] Compounds intended for administration to humans or other mammals generally should have very high purity. In the case of synthetically prepared compounds, purity refers to the ratio of a compound’s mass to the total sample mass following any purification steps. Usually, the level of purity is at least about 95%, more usually at least about 96%, about 97%, about 98%, or higher. For example, the level of purity may be about 98.5%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or higher.

[0084]

[0057] Compounds described herein that exist in more than one optical isomer form (enantiomers) may be provided either as racemic mixture or by isolating one of the enantiomers, the latter case in which purity as described above may refer to enantiomeric purity.

[0085] IV. Methods of use

[0086]

[0058] As described above, cannabinoids and related cannabinoid analogs typically exert therapeutic and anti-inflammatory activities via CB2 cannabinoid receptors. While not wanting to be bound by theory, compounds disclosed herein may also exhibit properties as inhibitors of monoamine oxidase (MAO) activity, including either or both of MAO- A and MAO-B activity. These properties may enable compounds to be effective for Atty DktNo. 009806.00123 treating indications associated with MAO activity, such as depression, pain, a sleep disorder, substance addiction, smoking cessation, and the like. Hence, in some embodiments, the compounds disclosed herein can be used in a method of treating diseases and conditions associated with monoamine oxidase (MAO) activity. In some embodiments, the individual suffers from depression, pain, a sleep disorder, or addiction.

[0087]

[0059] Compounds disclosed herein also (or alternatively) may exhibit anti-inflammatory properties owing to the compound’s interaction with inflammation pathways, including by way of example, interleukins such as IL-1 and IL-6, TNF-a, cyclooxygenase (COX), and the like. A compound’s ability to inhibit MAO-A and / or MAO-B activity, and / or its ability to inhibit COX and / or other pathways associated with inflammation may be evaluated using assays well known to persons of ordinary skill in the art.

[0088]

[0060] In some aspects, a cannabinoid analog as described herein is administered to an individual in need thereof for the treatment of a substance addiction, such as alcohol, tobacco, opioid, prescription drugs, cocaine, benzodiazepines, amphetamines, hallucinogens, inhalants, phencyclidine, or other drug addictions. Such treatments also are inclusive of treating withdrawal in dependency on benzodiazepines, opiates, or alcohol, as well as symptoms experienced by patients with substance use disorders, such as anxiety, mood symptoms, pain, and sleep disorders such as insomnia.

[0089]

[0061] In addition to anxiety that is associated with substance use disorders, the cannabinoid analogs may be effective for treating other types of anxiety disorders, such as post- traumatic stress disorder, general anxiety disorder, panic disorder, social anxiety disorder, and obsessive-compulsive disorder.

[0090]

[0062] In other aspects, a cannabinoid analog as described herein may be administered to an individual in need thereof for the treatment of multiple sclerosis, fibromyalgia, epilepsy or neuropsychiatric disorders that are linked to epilepsy, such as neurodegeneration, neuronal injury, and psychiatric diseases. The cannabinoid analogs may be effective for potentiating the anticonvulsant activity of other active agents such as phenytoin and diazepam. Atty DktNo. 009806.00123

[0091]

[0063] In still other aspects, a cannabinoid analog as described herein may be used in as an antipsychotic for treating patients with schizophrenia. The cannabinoid analogs also may be effective to reduce intraocular pressure, such as in the treatment of glaucoma.

[0092]

[0064] In yet other aspects, a cannabinoid analog as described herein may be administered to an individual in need thereof for the treatment of cancer. The cannabinoid analog may be effective to block cancer cells from spreading around the body and invading an area entirely; for suppressing the growth of cancer cells and / or promoting the death of cancer cells.

[0093]

[0065] The cannabinoid analogs as described herein may be useful in the treatment of Type 1 diabetes, which is caused by inflammation when the immune system attacks cells in the pancreas; as well as acne, which is caused, in part, by inflammation and overworked sebaceous glands on the body. The anti-inflammatory properties of the compounds may lower the production of sebum that leads to acne, including acne vulgaris, the most common form of acne.

[0094]

[0066] The cannabinoid analogs as described herein may be used to treat Alzheimer’s disease, and particularly to prevent the development of social recognition deficit in subjects when administered in the early stages of Alzheimer’s disease. In some aspects, the compounds may be administered to improve cognition. Other examples of disorders that may be treated by the cannabinoid analog as described herein include nausea, vomiting, anorexia, and cachexia. The compounds may produce an appetite -enhancing effect, for example in AIDS patients or individuals with Alzheimer’s disease who refuse food.

[0095]

[0067] The cannabinoid analogs as described herein may be useful in the treatment of spasticity caused by multiple sclerosis (MS) or spinal cord injury, movement disorders, such as Tourette’s syndrome, dystonia, or tardive dyskinesia. MS patients may experience benefits on ataxia and reduction of tremors.

[0096]

[0068] Analgesic properties of the cannabinoid analogs may prove beneficial, for example, in the treatment of neuropathic pain due to multiple sclerosis, damage of the brachial plexus and HIV infection, pain in rheumatoid arthritis, cancer pain, headache, menstrual pain, chronic bowel inflammation and neuralgias. Atty DktNo. 009806.00123

[0097]

[0069] The cannabinoid analogs as described herein may be useful in the treatment of asthma. Experiments examining the anti-asthmatic effect of THC or cannabis date mainly from the 1970s, and are all acute studies. The effects of a cannabis cigarette (2% THC) or oral THC (15 mg), respectively, approximately correspond to those obtained with therapeutic doses of common bronchodilator drugs (salbutamol, isoprenaline). Since inhalation of cannabis products may irritate the mucous membranes, oral administration or another alternative delivery system would be preferable. Very few patients developed bronchoconstriction after inhalation of THC.

[0098]

[0070] An improvement of mood in reactive depression has been observed in several clinical studies with THC. There are additional case reports claiming benefit of cannabinoids in other psychiatric symptoms and diseases, such as sleep disorders, anxiety disorders, bipolar disorders, and dysthymia. Various authors have expressed different viewpoints concerning psychiatric syndromes and cannabis. While some emphasize the problems caused by cannabis, others promote the therapeutic possibilities. Quite possibly cannabis products may be either beneficial or harmful, depending on the particular case. The attending physician and the patient should be open to a critical examination of the topic, and a frankness to both possibilities.

[0099]

[0071] In a number of painful syndromes secondary to inflammatory processes (e.g. ulcerative colitis, arthritis), cannabis products may act not only as analgesics but also demonstrate anti-inflammatory potential. For example, some patients employing cannabis report a decrease in their need for steroidal and nonsteroidal anti-inflammatory drugs. Moreover there are some reports of positive effects of cannabis self-medication in allergic conditions. It is as yet unclear whether cannabis products may have relevant effects on causative processes of autoimmune diseases.

[0100]

[0072] There are a number of positive patient reports on medical conditions that cannot be easily assigned to the above categories, such as pruritus, hiccup, ADS (attention deficit syndrome), high blood pressure, tinnitus, chronic fatigue syndrome, restless leg syndrome, and others. Different authors have described several hundred possible indications for cannabis and THC. For example, 2.5 to 5 mg THC were effective in three patients with pruritus due to liver diseases. Another example is the successful treatment Atty DktNo. 009806.00123 of a chronic hiccup that developed after a surgery. No medication was effective, but smoking of a cannabis cigarette completely abolished the symptoms.

[0101]

[0073] Cannabis products often show very good effects in diseases with multiple symptoms that encompassed within the spectrum of THC effects, for example, in painful conditions that have an inflammatory origin (e.g., arthritis), or are accompanied by increased muscle tone (e.g., menstrual cramps, spinal cord injury), or in diseases with nausea and anorexia accompanied by pain, anxiety and depression, respectively (e.g. AIDS, cancer, hepatitis C).

[0102]

[0074] COVID- 19 is transmitted through respiratory droplets and uses receptor-mediated entry into a human host via angiotensin-converting enzyme II (ACE2) that is expressed in lung tissue, as well as oral and nasal mucosa, kidney, testes, and the gastrointestinal tract. Modulation of ACE2 levels in these gateway tissues may decrease disease susceptibility. See Wang et al., In Search of Preventative Strategies: Novel Anti- Inflammatory High-CBD Cannabis Sativa Extracts Modulate ACE2 Expression in COVID- 19 Gateway Tissues (April 17, 2020), doi: 10.20944 / preprints202004.0315.vl. The cannabinoid analogs as described herein may modulate ACE2 expression and may have utility in the treatment of a coronavirus such as COVID- 19.

[0103] V. Dosage and pharmaceutical compositions

[0104]

[0075] Suitable doses may vary over a wide range depending on a variety of factors including the type and / or severity of the disease or disorder, previous treatments, the general health, age, and / or weight of the individual, the frequency of treatments, the rate of release from the composition, and other diseases present. This dose may vary according to factors such as the disease state, age, and weight of the subject. For example, higher doses may be administered for treatments involving conditions that are at an advanced stage and / or life threatening. Dosage regimens also may be adjusted to provide the optimum therapeutic response.

[0105]

[0076] Pharmaceutical compositions may be formulated together with one or more acceptable pharmaceutical or food grade carriers or excipients. As used herein, the term “acceptable pharmaceutical or food grade carrier or excipient” means a non-toxic, inert solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. For example, sugars such as lactose, glucose and sucrose; starches such as com Atty DktNo. 009806.00123 starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol, and phosphate buffer solutions, as well as compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0106]

[0077] Pharmaceutical compositions may be prepared by any suitable technique and is not limited by any particular method for its production. For example, purified cannabinoids can be combined with excipients and a binder, and then granulated. The granulation can be dry-blended with any remaining ingredients, and compressed into a solid form such as a tablet.

[0107]

[0078] Pharmaceutical compositions may be administered by any suitable route. For example, the compositions may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, via an implanted reservoir, or ingested as a dietary supplement or food. In some embodiments, a composition is provided in an inhaler, which may be actuated to administer a vaporized medium that is inhaled into the lungs. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, and intracranial injection or infusion techniques. Most often, the pharmaceutical compositions are readily administered orally and ingested.

[0108]

[0079] Pharmaceutical compositions may contain any conventional non-toxic pharmaceutically -acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation may be adjusted with acceptable pharmaceutical or food grade acids, bases or buffers to enhance the stability of the formulated composition or its delivery form.

[0109]

[0080] Liquid dosage forms for oral administration include acceptable pharmaceutical or food grade emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly Atty DktNo. 009806.00123 used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 -butylene glycol, dimethylsulfoxide (DMSO) dimethylformamide, oils (in particular, cottonseed, groundnut, com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0110]

[0081] Solid dosage forms for oral administration include capsules, tablets, lozenges, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, acceptable pharmaceutical or food grade excipient or carrier such as sodium citrate or dicalcium phosphate and / or a) fdlers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, c) humectants such as glycerol, d) disintegrating agents such as agaragar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof, and j) sweetening, flavoring, perfuming agents, and mixtures thereof. In the case of capsules, lozenges, tablets and pills, the dosage form may also comprise buffering agents.

[0111]

[0082] The solid dosage forms of tablets, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract or, optionally, in a delayed or extended manner. Examples of embedding compositions which can be used include polymeric substances and waxes. Tablet formulations for extended release are also described in U.S. Pat. No. 5,942,244. Atty DktNo. 009806.00123

[0112]

[0083] Compositions may contain a cannabinoid analog or compounds, alone or with other therapeutic compound(s). A therapeutic compound is a compound that provides pharmacological activity or other direct effect in the diagnosis, cure, mitigation, treatment, or prevention of disease, or to affect the structure or any function of the body of man or animals. A therapeutic compound disclosed herein may be used in the form of a pharmaceutically acceptable salt, solvate, or solvate of a salt, e.g., a hydrochloride. Additionally, therapeutic compound disclosed herein may be provided as racemates, or as individual enantiomers, including the R- or S-enantiomer. Thus, the therapeutic compound disclosed herein may comprise a R-enantiomer only, a S-enantiomer only, or a combination of both a R-enantiomer and a S-enantiomer of a therapeutic compound. In some aspects, the therapeutic compound may have anti-inflammatory activity, such as a non-steroidal anti-inflammatory drug (NS AID). NSAIDs are a large group of therapeutic compounds with analgesic, anti-inflammatory, and anti-pyretic properties. NSAIDs reduce inflammation by blocking cyclooxygenase. NSAIDs include, without limitation, aceclofenac, acemetacin, actarit, alcofenac, alminoprofen, amfenac, aloxipirin, aminophenazone, antraphenine, aspirin, azapropazone, benorilate, benoxaprofen, benzydamine, butibufen, celecoxib, chlorthenoxacin, choline salicylate, clometacin, dexketoprofen, diclofenac, diflunisal, emorfazone, epirizole; etodolac, etoricoxib, feclobuzone, felbinac, fenbufen, fenclofenac, flurbiprofen, glafenine, hydroxylethyl salicylate, ibuprofen, indometacin, indoprofen, ketoprofen, ketorolac, lactyl phenetidin, loxoprofen, lumiracoxib, mefenamic acid, meloxicam, metamizole, metiazinic acid, mofebutazone, mofezolac, nabumetone, naproxen, nifenazone, niflumic acid, oxametacin, phenacetin, pipebuzone, pranoprofen, propyphenazone, proquazone, protizinic acid, rofecoxib, salicylamide, salsalate, sulindac, suprofen, tiaramide, tinoridine, tolfenamic acid, valdecoxib, and zomepirac.

[0113]

[0084] NSAIDs may be classified based on their chemical structure or mechanism of action. Non-limiting examples of NSAIDs include a salicylate derivative NSAID, a p-amino phenol derivative NSAID, a propionic acid derivative NSAID, an acetic acid derivative NSAID, an enolic acid derivative NSAID, a fenamic acid derivative NSAID, a non- selective cyclooxygenase (COX) inhibitor, a selective cyclooxygenase- 1 (COX-1) inhibitor, and a selective cyclooxygenase -2 (COX-2) inhibitor. An NSAID may be a profen. Examples of a suitable salicylate derivative NSAID include, without limitation, acetylsalicylic acid (aspirin), diflunisal, and salsalate. Examples of a suitable p-amino Atty DktNo. 009806.00123 phenol derivative NSAID include, without limitation, paracetamol and phenacetin. Examples of a suitable propionic acid derivative NSAID include, without limitation, alminoprofen, benoxaprofen, dexketoprofen, fenoprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, loxoprofen, naproxen, oxaprozin, pranoprofen, and suprofen. Examples of a suitable acetic acid derivative NSAID include, without limitation, aceclofenac, acemetacin, actarit, alcofenac, amfenac, clometacin, diclofenac, etodolac, felbinac, fenclofenac, indometacin, ketorolac, metiazinic acid, mofezolac, nabumetone, naproxen, oxametacin, sulindac, and zomepirac. Examples of a suitable enolic acid (oxicam) derivative NSAID include, without limitation, droxicam, isoxicam, lomoxicam, meloxicam, piroxicam, and tenoxicam. Examples of a suitable fenamic acid derivative NSAID include, without limitation, flufenamic acid, mefenamic acid, meclofenamic acid, and tolfenamic acid. Examples of a suitable selective COX-2 inhibitors include, without limitation, celecoxib, etoricoxib, firocoxib, lumiracoxib, meloxicam, parecoxib, rofecoxib, and valdecoxib.

[0114]

[0085] A therapeutically effective amount of a therapeutic compound disclosed herein generally is in the range of about 0.001 mg / kg / day to about 100 mg / kg / day. An effective amount may be, e.g., at least 0.001 mg / kg / day, at least 0.01 mg / kg / day, at least 0.1 mg / kg / day, at least 1.0 mg / kg / day, at least 5.0 mg / kg / day, at least 10 mg / kg / day, at least 15 mg / kg / day, at least 20 mg / kg / day, at least 25 mg / kg / day, at least 30 mg / kg / day, at least 35 mg / kg / day, at least 40 mg / kg / day, at least 45 mg / kg / day, or at least 50 mg / kg / day. In some examples, an effective amount of a therapeutic compound may be in the range of about 0.001 mg / kg / day to about 10 mg / kg / day, about 0.001 mg / kg / day to about 15 mg / kg / day, about 0.001 mg / kg / day to about 20 mg / kg / day, about 0.001 mg / kg / day to about 25 mg / kg / day, about 0.001 mg / kg / day to about 30 mg / kg / day, about 0.001 mg / kg / day to about 35 mg / kg / day, about 0.001 mg / kg / day to about 40 mg / kg / day, about 0.001 mg / kg / day to about 45 mg / kg / day, about 0.001 mg / kg / day to about 50 mg / kg / day, about 0.001 mg / kg / day to about 75 mg / kg / day, or about 0.001 mg / kg / day to about 100 mg / kg / day. In other examples, an effective amount of a therapeutic compound disclosed herein may be in the range of, e.g., about 0.01 mg / kg / day to about 10 mg / kg / day, about 0.01 mg / kg / day to about 15 mg / kg / day, about 0.01 mg / kg / day to about 20 mg / kg / day, about 0.01 mg / kg / day to about 25 mg / kg / day, about 0.01 mg / kg / day to about 30 mg / kg / day, about 0.01 mg / kg / day to about 35 mg / kg / day, about 0.01 mg / kg / day to about 40 mg / kg / day, about 0.01 mg / kg / day to about 45 mg / kg / day, about 0.01 mg / kg / day to Atty DktNo. 009806.00123 about 50 mg / kg / day, about 0.01 mg / kg / day to about 75 mg / kg / day, or about 0.01 mg / kg / day to about 100 mg / kg / day.

[0115]

[0086] In addition to pharmaceutical compositions, compounds described herein may be formulated as an elixir, a beverage, a chew, a tablet, a lozenge, a gum, or the like. According to another aspect, the pharmaceutical compositions may also be formulated as a pharmaceutically acceptable vehicle such as a capsule, tablet, syrup, lozenge, inhaler, e-cigarette, chewable gum, nasal spray, transdermal patch, liquid, transmucosal vehicle, hydrogel, nanosome, liposome, noisome, nanoparticle, nanosphere, microsphere, microparticle, microemulsion, nanosuspension, or micelle. The compositions may also be formulated, for example, as dietary supplements or nutraceuticals.

[0116]

[0087] The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

[0117]

[0088] Specific elements of any of the foregoing embodiments can be combined or substituted for elements in other embodiments. Furthermore, while advantages associated with certain embodiments of the disclosure have been described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all Atty DktNo. 009806.00123 embodiments need necessarily exhibit such advantages to fall within the scope of the disclosure.

[0118] Example 1

[0119]

[0089] This example describes the synthesis of 3a-isopropyl-2-methyl-6-propyl-3a,8b-dihydro- lH-cyclopenta[b]benzofuran-8-ol (“T-55 ”) .

[0120] Synthesis of 4-Bromocyclopent-2-enone (2):

[0121]

[0090] To a solution of cyclopent-2-en-l-one (100 g, 1218 mmol) in anhydrous chloroform (1.50 L) under argon atmosphere at r.t, was added NBS (325 g, 1827 mmol) portion wise over a period of 30 minutes. After complete addition, the reaction mixture was refluxed at 80 °C (oil bath temp) for 48 h. The reaction mixture was cooled to r.t, diluted with water (2.0 L) and extracted with CH2CI2 (2 x 1.0 L). The combined organic extracts were washed with ice cold water (1.0 L), brine (1.0 L), dried over Na2SOr, filtered and concentrated under reduced pressure to obtain 4-bromocyclopent-2-en-l-one (150 g, 76%) as a yellow liquid.

[0122]

[0091] 'HNMR showed desired signals: 'HNMR (400 MHz, CDCh): d 7.67 (dd, J= 5.52, 2.56 Hz, 1H), 6.27 (dd, J = 5.52, 0.95 Hz, 1H), 5.23-5.06 (m, 1H), 3.05 (dd, J= 19.5, 6.28 Hz, 1H), 2.72 (dd, J= 19.5, 1.53 Hz, 1H). Atty DktNo. 009806.00123

[0123] Synthesis of 4-(prop-l-en-2-yl) cyclopent-2-en-l-one (3):

[0124]

[0092] To a stirred solution of LiCl (23.4 g, 558 mmol) in anhydrous THF (500 mL) was added CuCN (25.0 g, 279 mmol) and stirred for 20 min at room temperature. The reaction mixture was cooled to -40 °C and added isopropenyl magnesium bromide (279.5 mL, 0.5 M in THF, 139.7 mmol) dropwise over a period of 60 min, further stirred for 20 min at - 40 °C and then cooled to -78 °C. Then added 4-bromocyclopent-2-en-l-one (15 g, 93.1 mmol, 1.0 equiv) in anhydrous THF (100 mL) dropwise over a period of 60 min and the reaction mixture was stirred for 10 min at -78°C. The reaction mixture was slowly brought to 0 °C and quenched with saturated NHrCl solution (500 mL) and stirred for 30 min at room temperature. The reaction mixture was fdtered and the fdtrate was extracted with MTBE (2 x LO L). The combined MTBE extracts were washed with brine (2 x 1.0 L), dried over anhydrous Na2SOr, fdtered and concentrated under vacuum to obtain the crude product. The product was purified by silica gel chromatography (15- 25% MTBE in hexanes). The fractions containing the product were combined and concentrated under reduced pressure to obtain 4-(Prop-l-en-2-yl)cyclopent-2-en-l-one (4.0 g, 35%) as a colourless oil. Note: Compound 3 is volatile in nature, rotavapor bath temp should be < 20 °C, vacuum ~ 600 mbar.

[0125]

[0093] 1H NMR showed desired signals along with traces of MTBE solvent.1H NMR (400 MHz, CDCh): 37.60 (dd, 7= 5.65, 2.49 Hz, 1H), 6.24 (dd, 7= 5.54, 1.98 Hz, 1H), 4.90- 4.71 (m, 2H), 3.64-3.40 (m, 1H), 2.60 (dd, J = 18.8, 6.75 Hz, 1H), 2.18 (dd, J = 18.8, 2.02 Hz, 1H), 1.71 (s, 3H).

[0126] Synthesis of l-methyl-4-(prop-l-en-2-yl)cyclopent-2-en-l-ol (4):

[0127]

[0094] A solution of methyl Lithium (1.6 M in diethyl ether, 69.1 mL, 73.72 mmol) was added dropwise over a period of 30 min to a solution of 4-(Prop-l-en-2-yl)cyclopent-2-en-l- one (9.0 g, 73 mmol) in anhydrous THF (100 mL) at -78°C under argon atmosphere and the reaction mixture was stirred at -78 °C for 10 min. The reaction mixture was slowly brought to 0 °C, quenched with NH4CI solution (130 mL) and extracted with MTBE (2 x 500 mL). The combined MTBE extracts were washed with water (500 mL), brine (500 mL) and dried over Na2SC>4, fdtered and concentrated at reduced pressure to obtain 1- methyl-4-(prop-l-en-2-yl)cyclopent-2-en-l-ol (4.5 g, crude) as a light yellow liquid Atty DktNo. 009806.00123 which was directly used in the next step. Note: Compound 4 is also volatile in nature, rotavapor bath temp should be < 20 °C, vacuum ~ 600 mbar.

[0128] Synthesis of 2-(3-methyl-5-(prop-l-en-2-yl)cyclopent-2-en-l-yl)-5-propylbenzene- 1,3-diol (5):

[0129]

[0095] To a stirred solution of Brockmann 1 activated basic alumina (79 g, 782 mmol) in anhydrous CH2CI2 (300 mb) was added boron trifluoride diethyl etherate (13.4 mb, 108.6 mmol) at room temperature dropwise over a period of 20 min. The reaction mixture was slowly heated to 40 °C and stirred for another 10 min, followed by dropwise addition of a solution of 1 -methyl -4-(prop-l-en-2-yl)cy clopent-2 -en-l-ol (6.0 g, 138.1 mmol), 5-propylbenzene-l,3-diol (4.6 g, 30 mmol) in anhydrous CH2CI2 (100 mb). The reaction mixture was stirred at 40 °C for 5 min. The reaction mixture was cooled to room temperature and diluted with saturated NaHCO.i (300 mL) and extracted with CH2CI2 (300 mL x 2). The combined organic extracts were washed with water (500 mL), brine (500 mL), dried over Na2SOr, fdtered and concentrated under reduced pressure. The crude residue was purified twice by silica gel chromatography (6-10% EtOAc in hexanes). The fractions containing the pure product were combined and concentrated under reduced pressure to obtain 2-(3-methyl-5-(prop-l-en-2-yl)cyclopent-2-en-l-yl)-5- propylbenzene-l,3-diol (3.0 g, crude) as a brown liquid.XH NMR showed mixture of compound 5 and compound T-55.

[0130] Synthesis of 3a-isopropyl-2-methyl-6-propyl-3a,8b-dihydro-lH-cyclopenta[b] benzofuran-8-ol (T-55):

[0131]

[0096] To a solution of 2-(3-methyl-5 -(prop- l-en-2-yl)cy clopent-2 -en-l-yl)-5 -propylbenzene- 1,3-diol (3.0 g, 9.9 mmol) in anhydrous CH2CI2 (500 mL) was added Fe(OTf)3 (0.99 g, 1.98 mmol) and stirred at room temperature for 72 h. The reaction was cooled to 0 °C and quenched with saturated NaHCCh solution (100 mL) and extracted with CH2CI2 (2 x 100 mL). The combined CH2CI2 extracts were washed with water (100 mL), brine (100 mL), dried over anhydrous Na2SOr, filtered and concentrated under vacuum to obtain the crude product. The product was purified twice by silica gel chromatography (3-6% EtOAc in hexanes). The fractions containing the pure product were combined and concentrated under reduced pressure to obtain 2,4,4-trimethyl-7-propyl-3,3a,4,9b- tetrahydrocyclopenta[c]chromen-9-ol (086 g, 29 %) as a light brown liquid. Atty DktNo. 009806.00123

[0132]

[0097] 1H NMR showed desired signals: ' H NMR1H NMR (400 MHz, DMSO- Q: 9.20 (s, 1H), 6.06 (s, 1H), 5.96 (s, 1H), 5.34 (s, 1H), 3.64 (d, J= 6.8 Hz, 1H), 2.66-2.60 (m, 1H), 2.40-2.33 (m, 3H), 2.02-1.95 (m, 1H), 1.70 (s, 3H), 1.53-1.44 (m, 2H), 0.90-0.84 (m, 9H). MS (MM) m / z 273.1 [M + H]+.

[0133] Example 2 - Monoamine Oxidase Inhibitions

[0134]

[0098] Increasing concentration of test compounds was added to a reaction mixture containing the Monoamine Oxidase (MAO-B or MAO-A) enzyme to generate response curves. The reaction mixture used lOOmM potassium phosphate with a pH of 7.4 as incubation buffer. Spectrofluorimetric quantitation of 4-hydroxyquinoline was used to determine activity of the MAO-B or MAO-A enzyme.

[0135]

[0099] The assay was performed essentially as described in Biochem Pharmacol. 41(2): 155 - 162, and it was performed by Eurofins Panlabs, Inc. Where presented, ICso values were determined by a non-linear, least squares regression analysis using MathlQ™ (ID Business Solutions Ltd., UK). Where inhibition constants (Ki) are presented, the Ki values were calculated using the equation of Cheng and Prusoff (Cheng, Y ., Prusoff, W.H., Biochem. Pharmacol. 22:3099-3108, 1973) using the observed IC50 of the tested compound, the concentration of radioligand employed in the assay, and the historical values for the KD of the ligand (obtained experimentally at Eurofins Panlabs, Inc.). Where presented, the Hill coefficient (nH), defining the slope of the competitive binding curve, was calculated using MathlQ™. Hill coefficients significantly different than 1.0, may suggest that the binding displacement does not follow the laws of mass action with a single binding site.

[0136]

[0100] FIG. 1 shows the response curves of increasing concentration of 3a-isopropyl-2-methyl- 6-propyl-3a,8b-dihydro-lH-cyclopenta[b]benzofuran-8-ol (T-55) as indicated on the x- axis and percentage inhibition of MAO-B is shown on the y-axis. As shown in FIG. 1, the ICso for T-55 inhibition of MAO-B was determined to be 20.1 pM. The IC50 for reference compound R(-)-Deprenyl inhibition of MAO-B was determined to be 9.69 nM as shown in FIG. 1.

[0137]

[0101] Table 1 below show further experimental results for T-55 and Delta 9-THC. N.C. indicated in the table means “not calculated.” Aty DktNo. 009806.00123

[0138] Table 1

[0139]

[0102] Clorgyline and Deprenyl were used as reference compounds as shown in Table 2 below. Atty DktNo. 009806.00123

[0140] Table 2: Reference Compounds

[0141] Historical Concurrent

[0142] Cat# Assay Name Reference Compound ICso* Ki tin Batch * (CM*

[0143] Example 3

[0144]

[0103] This example describes the synthesis of 3a-isopropyl-2-methyl-6-pentyl-3a,8b-dihydro- lH-cyclopenta[b]benzofuran-8-ol (“M-55” or “MIRA-55”). Initially, compound 5 was prepared according to the following reaction scheme:

[0145] 4 5

[0146]

[0104] The target compound M-55 was prepared by cyclization of structure 5 under strongly acidic conditions, e.g., in the presence of trifluoromethanesulfonic acid (TFMS) or trimethylsilyl trifluoromethanesulfonate (TMSOTf). Compound 5 (11.5 g, 38.3 mmol, 1 eq) was dissolved in DCM (380 mL, 33 mL / g) and the solution was brought to 25°C by a water bath. TMSOTf (10.2 g, 46 mmol, 1.2 eq) was introduced in one portion and the reaction mixture was stirred at room temperature and monitored by Method #1 described below. After 2.5 hours the impurity profde did not improve. NaHCOi (400 mL) was added, and the phases were separated. The organic layer was washed with water, and brine, dried overNa2SO4, and evaporated to afford 12.3 g of the crude product as a brown oil. The crude was fdtered through a Silica gel pad (30% DCM in hexane (1 Atty DktNo. 009806.00123

[0147] L) and 50% DCM / hexane (0.5 L) to afford 7.1 g of a yellow oil, which was used in the next step without further purification.

[0148]

[0105] A series of trials exploring the influence of different cyclization conditions was carried out; the applied conditions are summarized in Table 2. The progress / conversion of the experiments was monitored by HPLC analytical Method # 1.

[0149] Table 2 Atty DktNo. 009806.00123

[0150]

[0106] Formation of M-55 was accompanied by formation of other isomers. Many isomers were formed at the initial stage, which mostly disappeared / transformed to the targeted structure after prolonged stirring (see FIG. 2). Note that the intensity of the product’s peak did not decrease over time, consistent with no product decomposition in the reaction mixture.

[0151]

[0107] After screening several reaction conditions (equivalents, reaction temperature, and type of addition) a final procedure was outlined and repeated on a larger scale. Trial 5 was monitored and aborted when no further improvement in the impurity profile was observed. The isolated crude product was combined with other small batches and purified by column chromatography to provide the product (trial 6) with 94.4% HPLC purity. An unknown impurity at RRT 1.09 was not separated. Since there was a significant difference in RT (HPLC) and visible difference in Rf (TLC) between the product and the impurity, it was assumed that tuning of the purification method would enable better resolution between them. The cyclization experiment was repeated (trial 7); however, use of a larger Silica-gel column and slower gradient provided again a mixture of M-55 and impurity RRT 1.09 (detailed in Table 2). Additional attempts to develop an appropriate chromatography method, implementing among the other methods purification on AgNCh impregnated Silica gel, showed no particular improvement. Atty DktNo. 009806.00123

[0152]

[0108] Fractions with high level of impurity RRT 1.09 were loaded on reverse phase preparative HPLC and several mg of the unknown impurity were isolated and submitted to NMR analysis. The structure of impurity RRT 1.09 was successfully interpreted and identified as the following structure:

[0153] RRT 1.09

[0154]

[0109] The impurity RRT 1.09 was removed by the hydroboration method depicted below. The experiments performed are summarized in Table 3.

[0155] Table 3 Aty DktNo. 009806.00123

[0156] [HO] Trials 9-13 were conducted using previously purified M-55 containing about 6% of impurity RRT 1.09. The batches were combined and purified by normal phase chromatography to provide 2.67 g of the product with 99.41% HPLC purity (0.18% impurity RRT 1.09, -18% isolated yield). Cyclization of M-5 was repeated (trial 16). The crude isolated product was filtered through a Silica pad and forwarded to the developed hydroboration method. Fractions with 97.94% HPLC purity (0.63% RRT 1.09) and 97.34% HPLC purity (0.52% RRT 1.09) were isolated, as summarized in Table 4. Trial 13 results for HPLC chromatogram, MS spectrum, 'H-NMR and13C- NMR are presented in FIGS. 3, 4, 5 and 6, respectively.

[0157] Table 4 Atty DktNo. 009806.00123

[0158] TMS Protection

[0159] [Hl] The previously isolated M-55 (trial 16) (7.1 g, 23.7 mmol, 1 eq) was dissolved in DCM (90 mL) and EtsN (7.2 g, 71 mmol, 3 eq) was added. The solution was cooled to 0-5°C and TMSC1 (3.9 g, 35.5 mmol, 1.5 eq) was added dropwise. The obtained reaction mixture was stirred at 0-5°C for 15 minutes, then allowed to heat to room temperature, stirred, and monitored by Method #2. After 1 hour a full conversion was detected, the reaction mixture was cooled to 0-5°C, and a white precipitate was fdtered (EhN^HCI). The fdtrate was evaporated to dryness, dissolved in hexane (100 mL), filtered, and evaporated again to afford 8 g of the protected M-55 as a yellow oil.

[0160] Hydroboration

[0161]

[0112] The previously isolated trial 11 (8 g, 21.5 mmol, 1 eq) was dissolved in THF (extra dry, 80 mL) and a solution of IM BHs’THF (6.5 mL, 6.5 mmol, 0.3 eq) was added dropwise at 0-5 °C. The obtained reaction mixture was stirred for 30 minutes at 0-5 °C, then allowed to warm to room temperature, stirred, and monitored by Method #1. After 1.5 hours the impurity RRT 1.09 decreased from 10% to 3%. An additional amount of 1 M BHi’THF (2 mL, 2 mmol, 0.1 eq) was added and the stirring was continued for 1 hour, the impurity RRT 1.09 decreased from 3% to 1.5%. The peak of impurity RRT 1.09 decreased from 3% to 1.5%. An additional amount of IM BHvTHF (1 mL, 1 mmol, 0.05 eq) was added and the stirring was continued for 1 hour, the impurity RRT 1.09 decreased from 1.5% to 0.7%. The reaction mixture was evaporated to dryness to afford 9 g of a crude yellow oil.

[0162] Deprotection

[0163]

[0113] The previously isolated trial 15 (9 g, 24.1 mmol, 1 eq) was dissolved in MeOH (100 mL) and TBAF (IM, 0.5 mL, 0.5 mmol, 0.02 eq) was added. The obtained reaction mixture was stirred at room temperature and monitored by Method #2 described below. After 1 hour full conversion was observed and the solvent was evaporated to dryness. The obtained residue was dissolved in hexane (16 mL), loaded on Silica gel column (220 g, 40-60 p) and eluted with 100% hexane (5 min), 0-30% DCM in hexane (10 min), 30% DCM in hexane (45 min) to afford 2.9 g of M-55 with 97.94% HPLC purity and 0.57 g of M-55 with 97.34% HPLC purity (total 30% yield). Trial 16 results for HPLC Atty DktNo. 009806.00123 chromatogram, MS spectrum, ' H-NMR and13C-NMRare presented in FIGS. 7, 8, 9 and 10, respectively.

[0164] Method #1 (analysis of M-55)

[0165] Column: Kinetex EVO C18, 1.7 p, 100x2.1 mm

[0166] Column temperature: 30°C

[0167] Flow rate: 0.5 mL / min

[0168] Injection volume: 1 pL

[0169] UV Detection: 220 nm, 250 nm

[0170] Mobile phase A: acetonitrile / MeOH=l / l

[0171] Mobile phase B: 0.1% H3PO4 in water

[0172] Diluent / Blank Solution: acetonitrile / water

[0173] Gradient: see Table 5 below

[0174] Table 5

[0175] Method #2 (monitoring of hydroboration method)

[0176] Column: XBridge C18, 3.5 p, 50x3 mm

[0177] Column temperature: 40°C

[0178] Flow rate: 1 mL / min

[0179] Injection volume: 2 p L

[0180] UV Detection: 220 nm, 254 nm, 305 nm

[0181] Mobile phase A: acetonitrile

[0182] Mobile phase B: lOmM ammonium carbonate in water

[0183] Diluent / Blank Solution: acetonitrile / water

[0184] Gradient: see Table 6 below Atty DktNo. 009806.00123

[0185] Table 6

[0186] Example 4 - Evaluation of antihyperalgesic effects of M-55 in inflammatory pain model

[0187]

[0114] Inflammatory pain is a complex pain condition that remains a significant clinical challenge due to the limited efficacy and / or poor tolerability of currently available treatment options. Inflammatory pain refers to spontaneous hypersensitivity to pain occurring in response to tissue damage and inflammation, including post-operative pain, traumatic pain, osteoarthritis pain, etc. This type of pathological pain state can be modeled using carrageenan-induced inflammation in which animals receive an intraplantar injection of carrageenan. Carrageenan is an irritating substance that induces transient inflammation and swelling that peaks 3-hr post injection and subsides by 24 hr. Carrageenan-induced inflammation results in hypersensitivity (hyperalgesia), which typically peaks after 180-min. THC has been shown to be effective at reducing carrageenan-induced hyperalgesia.

[0188]

[0115] The effects of M-55 were examined on carrageenan-induced hyperalgesia in male and female rats at Johns Hopkins University School of Medicine - Division of Behavioral Biology. M-55 or sesame oil vehicle was given 1-hr prior to carrageenan to determine effects on blocking or mitigating the induction of the inflammatory response. X- Carrageenan (Sigma Aldrich) was mixed in saline (1% w / v) and administered at a volume of 0.1 ml into the intra-plantar surface of one hind paw. Paw edema (swelling) was measured using an electronic digital caliper at the time of carrageenan injection (time 0) and 1, 3, and 5 hours post-injection, followed by measures of carrageenan- induced hyperalgesia with the Hargreaves and von Frey assays, which are plantar tests of thermal pain sensitivity and mechanical pain sensitivity, respectively. In the Hargreaves test, an infrared light generator is placed underneath the target hind paw Atty DktNo. 009806.00123

[0189] (midplantar area) and paw withdrawal latency is automatically recorded to the nearest 0.1 s. Latencies after carrageenan injection are compared with baseline measures recorded on the same day prior to treatment. Decreases in paw withdrawal latency indicate hyperalgesia (i.e., increased pain sensitivity / hypersensitivity), while increases in paw withdrawal latency indicate antinociception / analgesia (i.e., decreased pain sensitivity).

[0190]

[0116] In the von Frey test, the target hind paw is probed with von Frey filaments (9 filaments, 0.6-15.0 g, beginning with 2.0 g) on the plantar surface for 3s. The presence or absence of a response (nocifensive hind paw flexion reflex) is recorded. If no response occurs, a stronger stimulus is presented otherwise the next weaker stimulus is applied. This up- down process is repeated 4 times after the first change in response direction, and the 50% threshold for paw withdrawal is determined by the individual response pattern and the force of the last von Frey filament tested. Thresholds obtained after carrageenan injection are compared with baseline measures recorded on the same day prior to treatment. Decreases in paw withdrawal threshold indicate allodynia (i.e., increased pain sensitivity / hypersensitivity), while increases in paw withdrawal threshold indicate antinociception / analgesia (i.e., decreased pain sensitivity). Separate groups of animals were tested with oral THC (3, 10, mg / kg) or Sesame oil vehicle for comparison of effects with M-55. As a positive control, separate groups of animals were tested with the standard non-steroidal anti-inflammatory drug (NSAID) ketoprofen (10, 20 mg / kg, intraperitoneal administration, i.p.) or its vehicle (1: 1: 18, Ethanol: Cremophor:0.9% Saline; i.p.), given 60-minutes prior to carrageenan injection.

[0191]

[0117] Outcome measures were assessed as change from baseline test scores. Change in withdrawal latencies (Hargreaves; thermal pain sensitivity), change in withdrawal threshold (von Frey, mechanical pain sensitivity), and change in paw thickness (edema; measured in mm) were assessed. Outcomes were assessed with a three-way analysis of variance (ANOVA) with time as a within-subject factor and M-55 dose and sex as between subject factors. Two-way ANOVAs were conducted within each time point, planned a priori. Post hoc between-group comparisons were completed with Sidak’s or Dunnett’s tests. Atty DktNo. 009806.00123

[0192] Effects of M-55 on carrageenan-induced pain sensitivity and inflammation

[0193]

[0118] M-55 blocked thermal hyperalgesia (i.e., increased pain sensitivity) that is observed in the early phase of the inflammatory pain test. In an analysis of M-55 on the change in latency across the 3 timepoints (3-way ANOVA: sex, dose, and time as factors) (FIG. 1 la), there were significant main effects of time (F(2,148)=84.69, p<0.001) and sex (F(l, 74)=5.50, p=0.022). There was also a significant interaction of time x sex (F(l, 148)=4.12, p=0.018). There was a non-significant trend for an effect of dose (F(4, 72)= 2.21, p=0.076); interactions of time x dose and time x sex x dose were not significant (p’s>0.2).

[0194]

[0119] In analyses of each timepoint, a significant effect of dose (F(4, 83)= 2.76, p=0.034) and sex (F(l, 83)=8.77, p=0.004) was observed in the first hour. Post-hoc testing determined that 10 mg / kg M-55 treatment blocked hyperalgesia compared with Vehicle in this first hour (p=0.016; sexes collapsed) (FIG. 12). This effect was stronger in females (average change in latency: Vehicle: -5.43 s vs. 10 mg / kg M-55: 1.16s; p<0.05 vs Vehicle;), and while effects in males were not significant, results trended in a similar direction (Vehicle: -1.48s vs. 10 mg / kg M-55: 2.61s). No other doses tested were significantly different from Vehicle (p’s>0.05). Analyses of hours 3 and 5 did not indicate an effect of M-55 on thermal pain sensitivity at these time points. Therefore, treatment with 10 mg / kg of M-55 increased withdrawal latencies compared with vehicle at hour 1, indicating beneficial effects on inflammatory hyperalgesia.

[0195]

[0120] An analysis of all timepoints (3-way ANOVA: sex, dose, and time as factors) on the change in von Frey thresholds across the 3 timepoints did not indicate any significant effects of M-55 (FIG. 1 lb). There was no significant main effect of dose (F(4,74)=0.35, p=0.84), interaction of dose x time (F(8, 148)=.43, p=0.90), nor interaction of dose x sex x time (F(8, 148)=0.97, p=0.47). There was a significant interaction of dose x sex: F(4, 74)=2.98, p=0.02, indicating an impact of sex on M-55 response. Thus, we determined M-55 effects on mechanical allodynia separately in each sex. In males, there was no effect of M-55 on change in threshold (F(4, 38)= 1.37, p=0.26); in females, there was a non-significant trend for an effect of M-55 on change in threshold (F(4, 36)= 1.83, p=0.14). However, this effect appeared to be driven by effects of 100 mg / kg M-55 increasing mechanical allodynia in hour 1 compared with vehicle. Atty DktNo. 009806.00123

[0196]

[0121] Analysis of paw edema did not indicate any significant effects of M-55. In an analysis of all timepoints (3-way ANOVA: sex, dose, and time as factors) on paw edema across the 3 timepoints demonstrated no significant main effect of dose (F(4,74=0.18, p=0.95), interaction of dose x time (F(8, 148)=0.89, p=0.53), interaction of dose x sex (F(4, 74)=1.36, p=0.26), nor interaction of dose x sex x time (F(8, 148)=0.20, p=0.88). In analyses of each time point, there were no main effects of dose (p’s>0.05).

[0197]

[0122] In summary, M-55 was effective at blocking thermal hyperalgesia at the 1-hr timepoint. M-55 did not augment significantly mechanical pain sensitivity or inflammation at the doses tested.

[0198] Effects of THC on carrageenan-induced pain sensitivity and inflammation

[0199]

[0123] Overall, THC produced thermal and mechanical analgesia at moderate to high doses (3- 10 mg / kg), while a low dose (1 mg / kg) increased carrageenan-induced inflammation. THC increased withdrawal latencies in the Hargreaves test of thermal pain sensitivity (FIG. 13a). In an analysis of all timepoints (3-way ANOVA: sex, dose, and time as factors) on THC effects on the change in latencies across the 3 timepoints, there were significant main effects of time (F(2,120)=44.81, p<0.001) and dose (F(3,30)=5.75, p=0.002). There was no effect of sex (F(l,60)=0.45, p=0.50). There was a nonsignificant trend for an interaction of time x sex (F(2, 120)= 2.45, p=0.09); interactions of time x dose, sex x dose, and time x sex x dose were not significant (p’s>0.3). Post- hoc tests indicate that 3 and 10 mg / kg THC increased latencies compared to the vehicle condition.

[0200]

[0124] In analyses of each timepoint, there were significant main effects of dose in hour 1 (F(3, 60) = 3.61, p=0.02), hour 3 (F(3, 60) = 7.12, p<0.001), and hour 5 (F(3, 60) = 3.78, p=0.02). There were no significant effects of sex, or interaction of sex x dose within timepoints. For comparison with M-55 maximum effects in the Hargreaves test (Hour 1), we are showing effects of THC on analgesia in Hour 1 (p’s<0.02 for 3 and 10 mg / kg THC vs. Vehicle; sexes collapsed) (FIG. 14).

[0201]

[0125] THC increased withdrawal thresholds in the von Frey test of mechanical pain sensitivity test (FIG. 13b). In an analysis of THC effects on the change in threshold across all timepoints (3-way ANOVA: sex, dose, and time as factors), there were significant main Atty DktNo. 009806.00123 effects of dose (F(3,60)=16.58, p<0.001), time (F(2,120)=37.05, p<0.001), and a trend for a main effect of sex (F(l,60)=3.65, p=0.06). There were significant interactions of sex x dose: F(3, 60)=3.67, p=0.02, time x dose (F(6,120)=3.45, p<0.01), and a trend for an interaction of time x sex x dose (F(6, 120)=2.06, p=0.07). Post-hoc tests indicate that 3 and 10 mg / kg THC increased thresholds compared to the vehicle condition (p’s<0.001). THC was effective at increasing thresholds at all timepoints tested (1, 3, and 5 -hrs post-carrageenan), as indicated by significant main effects of dose within each timepoint (p’s<0.05).

[0202]

[0126] THC increased paw edema (FIG. 13c). In an analysis of all timepoints (3-way ANOVA: sex, dose, and time as factors) on THC effects on the change in paw thickness, there were significant main effects of dose (F(3,60)=5.66, p<0.01) and time (F(2,120)=95.35, p<0.001). There were significant interactions of time x sex (F(2,120)=5.28, p<0.01) and time x dose (F(6, 120)=4.56, p<0.01). Post-hoc tests indicate that 1 mg / kg THC tended to increase edema compared to the vehicle condition (p=0.054). In analyses of each time point, there was atrend for an effect of THC on edema at 1 hour (F(3,60)=2.67, p=0.06), and significant effects at hours 3 (F(3,60)=3.19, p=0.03) and 5 (F(3,60)=6.90, p<0.001).

[0203] Effects of ketoprofen on carrageenan-induced pain sensitivity and inflammation

[0204]

[0127] Overall, ketoprofen blocked hyperalgesia and reduced edema in a model of carrageenan- induced inflammatory pain. Ketoprofen significantly increased withdrawal latencies in the Hargreaves test of thermal pain sensitivity (FIG. 15a). In an analysis of all timepoints (3-way ANOVA: sex, dose, and time as factors) on ketoprofen effects on change in withdrawal latencies, there were significant main effects of dose: (F(2,58)=6.35, p<0.01) and time (F(2,116)=46.68, p<0.001). There were significant interactions of time x sex (F(2, 116)=5.41 , p=0.01 ) and time x dose (F(4, 116)=3.40, p=0.01 ) .

[0205]

[0128] In separate analyses of each time point, there were significant main effects of dose in Hour 3 (F(2,58) = 5.75, p<0.01) and Hour 5 (F(2,58) = 11.06, p<0.001). Effects observed at the Hour 3 timepoint were driven by effects in females, where both 10 and 20 mg / kg ketoprofen were effective at blocking hyperalgesia (p’s<0.05). Five hours after carrageenan, ketoprofen (10, 20 mg / kg) was effective at blocking hyperalgesia in both sexes (p’s<0.05). For comparison with M-55 maximum effects in the Hargreaves test Atty DktNo. 009806.00123

[0206] (Hour 1), we are presenting effects of ketoprofen maximum effects (Hour 5) with sexes collapsed (FIG. 16; main effect of dose: F (2, 61) = 10.56, p<0.001).

[0207]

[0129] Ketoprofen had modest effects on allodynia observed after carrageenan injection (FIG. 15b). In an analysis of ketoprofen effects on change in von Frey thresholds (mechanical pain sensitivity), there was a main effect of time (F(2, 116)=5.23, p=0.01) and an interaction of time x sex (F(2,l 16)=3.76, p=0.03). In analyses within each timepoint, there was a main effect of dose in Hour 3 (F(l,58)=3.56, p=0.04), where 10 mg / kg ketoprofen did reduce allodynia compared with vehicle (p=0.02; sexes collapsed), an effect mainly driven by females.

[0208]

[0130] Ketoprofen reduced paw edema (FIG. 15c). In an analysis of all timepoints (3-way ANOVA: sex, dose, and time as factors) on ketoprofen effects on the change in paw thickness, there were significant main effects of time (F(2,l 16)=67.95, p<0.001) and a trend for an overall main effect of dose (F(2,58)=2.53, p=0.09). There were interactions of time x dose (F(4,l 16)=7.85, p<0.001) and time x sex x dose (F(4,l 16)=2.77, p=0.03). In separate analyses of each time point, there was a significant main effect of dose in Hour 5 only (F(2,58)=6.16, p<0.01). Post-hoc tests indicate that 10 (males only) and 20 mg / kg Ketoprofen (both sexes) reduced edema compared with vehicle controls.

[0209]

[0131] Overall, 10 and 20 mg / kg ketoprofen were effective at blocking thermal hyperalgesia associated with inflammation in later time points (hours 3-5) and reduced paw edema (hour 5).

[0210] Conclusions

[0211]

[0132] M-55 was effective at preventing thermal hyperalgesia associated with inflammation in the early phase of the rodent carrageenan model. The early time-point specific effect suggest that this compound may be short-acting. Future testing would need to evaluate repeated or chronic dosing to achieve steady state blood levels.

[0212]

[0133] Oral administration of THC was effective at producing thermal and mechanical analgesia at the doses tested in the inflammatory pain model of carrageenan. These effects were evident across all time points. The low dose of THC (1 mg / kg), which did not modulate pain sensitivity, increased edema. The antihyperalgesic effects of 10 mg / kg Atty DktNo. 009806.00123

[0213] THC and 10 mg / kg M-55 were equivalent (Mean difference from vehicle: +5.7s and +5 ,4s, respectively). However, effects of oral THC on hyperalgesia was long-lasting, with effectiveness observed until the last time point tested (5 hours after carrageenan).

[0214]

[0134] Ketoprofen was an effective analgesic, blocking thermal hyperalgesia at hours 3-5 following carrageenan-induced inflammation. At 3 hours, ketoprofen modestly reduced allodynia, though this effect was primarily driven by females. Ketoprofen was also effective at reducing edema in hour 5. This agrees with other reports4 and its mechanism of action 10. In comparison with M-55, effects of ketoprofen on thermal hyperalgesia occurred in the later phase of inflammatory pain sensitization. Further, ketoprofen (10, 20 mg / kg) showed effectiveness in reducing edema, while M-55 did not reduce edema.

[0215] Evaluation of the acute antinociceptive and cataleptic effects of M-55 at multiple time points

[0216]

[0135] Analgesic effects of drugs are commonly determined using assays measuring changes in pain sensitivity. Acute pain sensitivity was measured at baseline (-30 before drug administration) and again +30, +60, and +120 minutes following oral administration of M-55 (3, 10, 30, 100 mg / kg) or vehicle (Sesame oil) with the tail flick and von Frey assays. The tail flick assay and von Frey assay are standard tests to assess acute pain sensitivity with demonstrated sensitivity to analgesic and antihyperalgesic drugs, including cannabinoids. Conducting multiple assessments across a 2-hr duration will inform the time course and duration of drug action. Outcome measures will be withdrawal latencies and thresholds for tail flick and von Frey tests, respectively. We also assessed catalepsy in conjunction with acute pain sensitivity to determine whether M-55 produced classical effects of cannabinoid-1 receptor (CB1) activation. CB1 agonists, including THC 15, cause prototypical antinociception as well as catalepsy. Catalepsy was measured across 3 trials using the bar test, where rats are placed in an abnormal posture with their front paws on a bar (5 cm in diameter) that is 12-cm high. Outcome measure is the time in seconds that paws maintain contact with the bar, with a maximum test time of 180-s. Catalepsy was tested +150 minutes following oral M-55, as THC -induced catalepsy is most evident at later time points. Catalepsy is expressed as the In(sec) of the average across 3 trials.

[0217]

[0136] Pain sensitivity outcome measures were assessed as change from baseline test scores. We assessed change in withdrawal latencies (tail flick; thermal pain sensitivity), change Atty DktNo. 009806.00123 in withdrawal threshold (von Frey, mechanical pain sensitivity). Catalepsy was assessed as total seconds in contact with the bar. Outcomes were assessed with a three-way analysis of variance (ANOVA) with time and dose as within-subject factors and sex as between subject factors. Two-way ANOVAs were conducted within each time point, plarmed a priori. Post hoc between-group comparisons were completed with Sidak’s or Dunnett’s tests.

[0218] Effects of M-55 on acute pain sensitivity and catalepsy.

[0219]

[0137] M-55 produced thermal and mechanical antinociception. M-55 increased latencies in the tail flick test of thermal pain sensitivity. Across all timepoints (3-way ANOVA), an analysis of change in tail flick withdrawal latencies indicated a significant main effect of dose (F(4,50)= 3.30, p=0.02) and a trend for an interaction of time x sex x dose (F(8, 100)= 1.83, p=0.08). Analysis within each timepoint indicates that M-55 increased latencies compared with vehicle when tested 60-minutes after administration (main effect of dose: F(4,50)= 4.35, p=0.004; FIG. 18a). Post-hoc tests show that rats tested with 100 mg / kg M-55 had higher latencies than those in the vehicle group (p<0.04). M- 55 did not significantly affect thermal pain when tested at 30-min (main effect of dose: F(4,50)= 1.78, p=0.15), and tended to increase latencies when tested at 120-min (main effect of dose: F(4,50)= 2.26, p=0.08).

[0220]

[0138] M-55 increased thresholds in the von Frey test of mechanical pain sensitivity. Across all timepoints (3-way ANOVA), an analysis of change in von Frey withdrawal thresholds indicated a significant interaction of time x sex x dose (F(8,100)= 2.08, p=0.05). Analysis within each timepoint indicates that M-55 increased thresholds compared with vehicle when tested 30-minutes after administration (main effect of dose: F(4,50)= 3.36, p=0.02; FIG. 18b). There were no effects of M-55 on mechanical pain sensitivity in the 1-hr and 2-hr time points post-drug administration.

[0221]

[0139] Catalepsy was not observed after administration with M-55 or vehicle (average time on bar <10s average; Mean ±SEM, Vehicle: 4.73 ±0.3.2 s, 3 mg / kg M-55: 1.18 ±0.30 s; 10 mg / kg M-55: 3.83 ±2.53 s; 30 mg / kg M-55: 3.97 ±2.87 s; 100 mg / kg M-55: 1.58 ±0.09 s data not shown). Atty DktNo. 009806.00123

[0222] Conclusions

[0223]

[0140] Acute administration of 100 mg / kg M-55 was effective at producing antinociception in normal rats. These effects were limited to early timepoints (30-min, 60-min), indicating this drug may be short acting. THC has also been shown to produce antinociception. In contrast to M-55 effects, our prior studies demonstrate that oral THC produces long lasting antinociception, with effects observed up to 5-hrs post-drug administration.

[0224]

[0141] Taken together with results from the inflammatory pain study, it can be concluded that M-55 has modest antinociceptive and anti -hyperalgesia effects that are limited to early time points. Different doses may be effective for acute vs. inflammatory pain. Future testing may evaluate repeated or chronic dosing to achieve steady-state blood levels and determine if greater and / or longer-lasting effects may be observed.

[0225] Example 5 - Evaluation of Anxiolytic Effects of M-55

[0226]

[0142] The Elevated Plus Maze (EPM) is a widely used preclinical behavioral assay for rodents and it has been validated to assess the anti-anxiety effects of pharmacological agents. EPM measures anxiety in rodents as a screening test for putative anxiolytic compounds and a general research tool in neurobiological anxiety research such as GAD or PTSD. The model is based on the test animal’s aversion to open spaces in the maze’s open arms.

[0227]

[0143] Anti-anxiety effects of test agents are demonstrated by an increase in the percentage of time spent in the open arm with treatment compared to placebo. The total distance traveled is a measure of the overall level of arousal and mobility of the mice undergoing testing on the EPM, and is used to rule out any sedating or intoxicating effects of the test agent. A number of clinically approved pharmacological agents to treat pain have been demonstrated to delay the onset of heat sensitivity upon paw exposure in mice to heat including opioids.

[0228]

[0144] The effect of acute administration of M-55 was studied on anxiety-related phenotypes in mice to model human conditions. An intraperitoneal (i.p.) injection of vehicle (saline + 1%DMSO) or MIRA-55 (0 mg / kg = Placebo [PBO] vs 50mg / kg = Treatment) was administered to 8-12-week-old C57B1 / 6 mice (n=5 / group). Thirty minutes following injection, mice were tested in anxiety-related measures using the EPM. As shown in Atty DktNo. 009806.00123

[0229] FIG. 19, at the doses tested M-55 has potent anti -anxiety effects without any symptoms of sedation or intoxication.

[0230]

[0145] While the invention has been described with respect to specific examples, those skilled in the art will appreciate that there are numerous variations and permutations of the above described systems and techniques that fall within the spirit and scope of the invention as set forth in the appended claims.

Claims

Atty DktNo. 009806.00123WHAT IS CLAIMED IS:

1. A compound having a structure of Formula (I) :wherein Ri and R2 are each independently selected from the group consisting of H, OH, protected hydroxyl, alkyl, alkenyl, alkynyl, acyl, aryl, heteroaryl, cycloalkyl, and heterocycle; wherein the alkyl, alkenyl, alkynyl or acyl is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRARB, — S-alkyl, — SO-alkyl, — SO2-alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, and heterocycle; wherein RAand RBare each independently selected from hydrogen and C1-4 alkyl; wherein the aryl or heteroaryl, whether alone or as part of a substituent group, is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, — COOH, — C(O)— Ci-4 alkyl, — C(O)O— Ci-4 alkyl, NRCRD, —S-alkyl, —SO-alkyl and — SO2-alkyl; wherein Rcand RDare each independently selected from hydrogen and C1-4 alkyl;R3 is selected from the group consisting of H, alkyl, acyl, — SO2-alkyl, — SO2-aryl and — SO2- heteroaryl; wherein the alkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogen, — OH, alkyl, — O-alkyl, NRERF, — S-alkyl, — SO-alkyl, — SO2-alkyl, aryl and heteroaryl; and wherein REand REare each independently selected from hydrogen and C1-4 alkyl; wherein the aryl or heteroaryl, whether alone or as part of a substituent group, is optionally substituted with one or more substituents independently selected from the group consisting of halogen,Atty DktNo. 009806.00123— OH, alkyl, — O-alkyl, NRGRH, — S-alkyl, — SO-alkyl and — SO2-alkyl; wherein RGand RHare each independently selected from hydrogen and C1-4 alkyl; and each - represents a single or double bond, with the proviso that within a 5 -membered ring, one or two - is a double bond and the remaining ->> are single bonds; or a pharmaceutically acceptable salt or ester thereof.

2. The compound of claim 1 wherein R2 is C1-C4 straight-chained or branched alkyl.

3. The compound of claim 1 wherein R2 is C5-C10 straight-chained or branched alkyl.

4. The compound of claim 1 which has a structure of Formula (IA):wherein one - is a single bond and the other ->> is a double bond; or a pharmaceutically acceptable salt or ester thereof.

5. The compound of claim 4 wherein R2 is C1-C4 straight-chained or branched alkyl.

6. The compound of claim 4 wherein R2 is C5-C10 straight-chained or branched alkyl.

7. A compound of claim 1 which has a structure of Formula (II):Atty DktNo. 009806.00123Formula (II) or a pharmaceutically acceptable salt or ester thereof.

8. The compound of claim 5 wherein R2 is C1-C4 straight-chained or branched alkyl.

9. The compound of claim 5 wherein R2 is C5-C10 straight-chained or branched alkyl.

10. The compound of claim 8 which has the structure:or a pharmaceutically acceptable salt or ester thereof.

11. The compound of claim 9 which has the structure:Atty DktNo. 009806.00123or a pharmaceutically acceptable salt or ester thereof.

12. A pharmaceutical composition comprising a compound of any preceding claim and a pharmaceutically acceptable vehicle therefor.

13. Use of a pharmaceutical composition of claim 12 for treating a cancer, tumor, addiction, epilepsy, Alzheimer’s disease, pain or depression.

14. Use of a pharmaceutical composition of claim 12 for treating anxiety, addiction, depression, a sleep disorder, or post-traumatic stress disorder (PTSD).

15. Use of a pharmaceutical composition of claim 12 for treating a cognitive disorder or for improving cognition.

Images