AU2018302155B2

AU2018302155B2 - Compositions containing cannabinoid analog conjugates and methods of use

Info

Publication number: AU2018302155B2
Application number: AU2018302155A
Authority: AU
Inventors: Jerry L. Bryant Jr.; Jana Rauvolfova; Tori STRONG; David J. Yang
Original assignee: Vyripharm Enterprises LLC
Current assignee: Vyripharm Enterprises LLC
Priority date: 2017-07-18
Filing date: 2018-07-18
Publication date: 2022-09-15
Anticipated expiration: 2038-07-18
Also published as: MX2020000673A; AU2018302155A1; CA3070538A1; EA202090116A1; EP3654971A1; WO2019018536A1; KR20200031663A; IL272060A; EP3654971A4; JP2020527589A; CN111065391A

Abstract

Provided here are compositions containing a conjugate of a label, a chelator, and a cannabinoid analog and methods for diagnosing, treating, or monitoring the progression of a cancer or a neurologic disorder using these compositions. Also provided here are methods of synthesizing these compositions and kits for delivery of these compositions as imaging and therapeutic agents.

Description

COMPOSITIONS CONTAINING CANNABINOID ANALOG CONJUGATES AND

METHODS OF USE

Cross-Reference to Related Application

[0001] This application claims the priority to and the benefit of U.S. Provisional Application No. 62/533,894, filed July 18, 2017, the contents of which are incorporated by reference in its entirety.

Field

[0002] This disclosure relates to synthetic cannabinoid compositions and methods generally directed to using such compositions for diagnostic, therapeutic, and prognostic applications.

Background

[0003] Medical cannabinoids have not been well developed for theranostic applications. Strategic clinical trials still lag behind, due to public stigmatization and law-imposed criminalization. However, petitions for therapeutic application of medical cannabinoids from healthcare providers, patients and activist advocates have exponentially increased. The cannabinoid receptors (CBi and CB₂), transient receptor potential subfamily V member 1 (TRPVl), transient receptor potential subfamily A member 1 (TRPAl), and the orphan receptor GPR55 are part of the endocannabinoid system (ECS), which is a complex lipid signaling network involving different proteins for the control or modulation of numerous physiological and pathophysiological processes. Along with these receptors, the ECS includes arachidonic acid- derived ligands, anandamide and 2-arachidonoyl glycerol, and the enzymes that degrade these endocannabinoids, such as fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL). While the expression of the CBi receptor is ubiquitous, with predominant presence in the brain, particularly in the basal ganglia, hippocampus, cerebellum and cortex, the CB₂ receptor is preferentially located in tissues that comprise the immune system (especially within the B cell rich compartments). Consistent with its broad distribution, CBi can be similarly detected in peripheral nerves and beyond the neural compartments, i.e., testis, uterus, vascular endothelium, eye, spleen, ileum and adipocytes. Because CBi and CB₂ receptors share 68% amino acid sequence identity at the trans-membrane domain, cannabinoid ligand/receptor pathways are being consistently evaluated as rational therapeutic targets for neurodegeneration in Alzheimer's, Parkinson's, Huntington's diseases; indolent pain, which is common in glaucoma and multiple sclerosis; and diverse cardiovascular disorders; and cancer.

Summary

[0004] Several disadvantages were recognized by the inventors and various embodiments of this disclosure were developed to address these shortcomings in the art. Certain embodiments disclosed and described here include compositions containing a conjugate of a label, a chelator, and a cannabinoid analog. The label can be a radionuclide. The label can be one or more of Technetium-99, Gallium-68, Copper-60, Copper-64, Indium-I l l, Holmium-166, Rhenium-186, Rhenium -188, Yttrium-90, Lutetium-177, Radium-223, or Actinium- 225. In certain embodiments, the label is configured to facilitate contrast-enhanced imaging, when the composition is administered to a mammal during use. The cannabinoid analog can be an immune check point cannabinoid receptor ligand. The cannabinoid analog can be a synthetic cannabinoid receptor agonist or a natural cannabinoid receptor agonist. The cannabinoid analog can be an aminoalkylindole. The cannabinoid analog can be a cannabinoid receptor inverse agonist. The cannabinoid analog can be a member of the group consisting of anandamide (AEA), 2- arachidonoylglycerol (2- AG), noladin ether, virodhamine and N-arachidonylodopamine (NAD A). The cannabinoid analog can be a diarylopyrazole. The chelator can an aminated or an acid chelator. In certain embodiments, the chelator is a cyclam. The cannabinoid analog can be cyclam-l'-acetyl- [N-(Piperidin-l-yl)-5-(4-chlorophenyl)-l-(2,4-dichlorophenyl)-4-methyl-lH-pyrazole-3- carboxamide] . The cannabinoid analog can be cyclam- 1 '-propyl- [N-(Piperidin-l -yl)-5-(4- chlorophenyl)- 1 -(2,4-dichlorophenyl)-4-methyl- lH-pyrazole-3-carboxamide] .

[0005] Embodiments also include methods for diagnosing a medical condition in a human subject, by administering to a human subject in need thereof an effective amount of any of the compositions described herein. A method of imaging a plurality of cancer cells, the method comprising administering to a human patient an effective amount of any of the compositions described herein. Embodiments also include methods for treating a cancer amenable to treatment with a cannabinoid, the method comprising administering to a human subject in need thereof an effective amount of any of the compositions described herein. Embodiments also include methods for treating a neurologic disorder amenable to treatment with a cannabinoid, the method comprising administering to a human subject in need thereof an effective amount of any of the compositions described herein. Embodiments also include methods for alleviating pain in a human subject, the method comprising administering to a human subject in need thereof an effective amount of the composition of any of the compositions described herein. Embodiments also include kits for imaging cells, comprising a predetermined quantity of a conjugate of a chelator and a medical cannabinoid analog; and a predetermined quantity of an imaging agent. The conjugate of the chelator and the medical cannabinoid analog can be present in the kit as precursors that subsequently interact with the imaging agent, when provided with suitable reaction conditions. The kit can include a tin-containing reducing agent. Cells, organs, and tissues that can be imaged using the kits include those of the cardiovascular system, precancerous cells and tissues, cancerous cells and tissues, and cells and tissues of the nervous system, including the brain and the spinal cord.

[0006] Numerous other aspects, features and benefits of the present disclosure may be made apparent from the following detailed description taken together with the drawing figures. The systems can include less components, more components, or different components depending on desired analysis goals.

Brief Description of the Drawings

[0007] While this disclosure is susceptible to various modifications and alternative forms, specific embodiments are shown by way of example in the drawings and will be described in detail here. The drawings may not be to scale. It should be understood, however, that the drawings and the detailed descriptions thereto are not intended to limit the disclosure to the particular form disclosed, but, to the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the scope of the present disclosure as defined by the appended claims.

[0008] FIG. 1 is a representative analysis of starting material SR141716 (rimonabant) using NMR spectroscopy, according to an embodiment.

[0009] FIG. 2 is a representative analysis of starting material SR141716 using high- performance liquid chromatography (HPLC) analysis, according to an embodiment.

[0010] FIG. 3 is a representative analysis of VYR206 using NMR, according to an embodiment.

[0011] FIG. 4 is a representative analysis of VYR206 using mass spectrometry, according to an embodiment. [0012] FIG. 5 is a representative analysis of VYR206 using HPLC analysis, according to an embodiment.

[0013] FIGS. 6A - 6C show the protein expression of the CBi receptor using Western blot analysis. FIG. 6A panel shows expression of the CBi receptor in cell lysate from various DLBCL. FIG. 6B shows expression of β-actin as the control for sample loading. FIG. 6C is a graphical representation of the mRNA expression analysis of CBi and CB₂ in various DLBCL cancer cell lines.

[0014] FIG. 7 is a graphical representation of the efficacy of cannabinoid receptor ligand SR141716 (rimonabant) on diffuse large B-cell lymphoma (DLBCL) cell lines' viability.

[0015] FIG. 8 is a graphical representation of the efficacy of cannabinoid receptor ligand CP945,598 (Otenabant) on DLBCL cell lines' viability.

[0016] FIG. 9 is a graphical representation of the efficacy of cannabinoid receptor ligand AM1241 on DLBCL cell lines' viability.

[0017] FIG. 10 is a graphical representation of the efficacy of cannabinoid receptor ligand AM251 on DLBCL cell lines' viability.

[0018] FIGS. 11A - 11D are graphical representations of the inhibition efficacy of two CBi inverse agonist SR141716 and VYR206 using DLBCL (FIGS. 11, A and B) and mantle cell lymphoma (MCL) cell lines (FIGS. 11, Panels C and D).

[0019] FIGS. 12A - 12D are graphical representations of the inhibition efficacy response of two selected CBi inverse agonist SR141716 and VYR206 and N4 using cell viability assays on DLBCL cancer cells: LR, MS, RC and DOHH2.

[0020] FIGS. 13A - 13D are graphical representations of the inhibition efficacy response of two selected CBi inverse agonist SR141716 and VYR206 and N4 using cell viability on MCL cancer cells: Jeko, Mino, Rec-1 and JMP1.

[0021] FIGS. 14A - 14C are graphical representations of the high through put screening assessment of cell viability in 16 DLBCL (FIGS. 14 A and B) and 8 MCL (FIG. 14C) cancer cell lines using cannabidiol, which was obtained as CBD in a coconut oil extract.

[0022] FIGS. 15A - 15D show the expression of CBi and CB₂ RNA in MCL and DLBCL cancer cell lines treated with VYR206 and CBD using immunoblot analysis. FIG. 15A panel shows expression of CBi with treatment with VYR206. FIG. 15B panel shows expression of CB₂ with treatment with VYR206. FIG. 15C panel shows expression of CB₂ with treatment with CBD. FIG. 15D panel shows expression of β-actin as the control for sample loading.

[0023] FIGS. 16A - 16B are graphical representations of the efficacy of a cannabidiol (CBD- 99) on MCL and DLBCL cell lines' viability. FIG. 16A is a graphical representation of the efficacy of CBD on MCL cell lines. FIG. 16B is a graphical representation of the efficacy of CBD on DLBCL cell lines.

[0024] FIG. 17 is a graphical representation of IC50 of rimonabant (identified as Rimo) versus Bruton's tyrosine kinase inhibitor - ibrutinib (Π3Ν) on various DLBCL cell lines.

[0025] FIGS. 18A - 18D are graphical representations of the effect of the cannabinoid receptor ligand - rimonabant (identified as Rimo) and a Bruton's tyrosine kinase inhibitor - ibrutinib (identified as IBN) on the viability of MCL cell lines (Jeko, Jeko-R, Mino, Mino-R, PF-4 and PF- 5) cell lines' viability. FIG. 18A is a graphical representation of the effect of 25 μΜ of rimonabant and ibrutinib on Jeko and Jeko-R cell lines. FIG. 18B is a graphical representation of the effect of 25 μΜ of rimonabant and ibrutinib on Mino and Mino-R. FIG. 18C is a graphical representation of efficacy of ibrutinib on PF-4 and PF-5 cell lines. FIG. 18D is a graphical representation of the effect of 25 μΜ of rimonabant and ibrutinib on PF-4 and PF-5.

[0026] FIGS. 19A - 19H are graphical representations of the efficacy of rimonabant and its cyclam conjugated form, VYR206 on the viability of DLCBL cell lines (LR, MS, RC and DOHH2) and MCL cell lines (Jeko, Rec-1, Mino and JMP-1). FIG. 19A is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of LR cell line with cyclam as a control. FIG. 19B is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of MS cell line with cyclam as a control. FIG. 19C is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of RC cell line with cyclam as a control. FIG. 19D is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of DOHH2 cell line with cyclam as a control. FIG. 19E is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of Jeko cell line with cyclam as a control. FIG. 19F is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of Rec-1 cell line with cyclam as a control. FIG. 19G is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of Mino cell line with cyclam as a control. FIG. 19H is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of JMP- 1 cell line with cyclam as a control. [0027] FIGS. 20A - 20D are graphical representations of the effect of a cannabidiol (CBD-99) on the viability of DLBCL (RC) and MCL (Mino) cell lines, individually and in combination with four chemo therapeutics: Bruton's tyrosine kinase inhibitors - ibrutinib (IBN) and zanubrutinib (BGB); proteasome inhibitor - carfilzomib (CFZ); and chemotherapeutic agent - tumorex (TMX). FIG. 20A is a graphical representation of the effect of 25 μΜ of CBD-99 and IBN, individually and in combination, on the viability of Mino and RC cells. FIG. 20B is a graphical representation of the effect of 6 μΜ of CBD-99 and 10 nM of CFZ, individually and in combination, on the viability of Mino and RC cells. FIG. 20C is a graphical representation of the effect of 25 μΜ of CBD and BGB, individually and in combination, on the viability of Mino and RC cells. FIG. 20D is a graphical representation of the effect of 12.5 μΜ of CBD and 250 nM of TMX, individually and in combination, on the viability of Mino and RC cells.

[0028] FIGS. 21A - 21E show the expression of cleaved PARP (cPARP) and cleaved caspase- 3 expression with increasing treatment of cannabidiol and VYR206 to demonstrate activation of apoptosis using immunoblot analysis. FIG. 21A panel shows expression of cPARP with increasing levels of cannabidiol and VYR206. FIG. 21B panel shows expression of β-actin as the control for sample loading. FIG. 21C panel shows expression of cPARP with increasing concentration of cannabidiol. FIG. 21D panel shows expression of cleaved caspase-3 with increasing concentration of cannabidiol. FIG. 21E panel shows expression of β-actin as the control for sample loading.

[0029] FIGS. 22A - 22B are graphical representations of the efficacy of cannabidiol, VYR206 (identified as N4-Rimo) and the combination of cannabidiol and VYR206 on the viability of DLCBL cell lines. FIG. 22A is a graphical representations of the efficacy of cannabidiol, VYR206, and the combination of cannabidiol and VYR206 on the viability of DLCBL cell line - LY19. FIG. 22B is graphical representations of the efficacy of cannabidiol, VYR206 (identified as N4-Rimo) and the combination of cannabidiol and VYR206 on the viability of DLCBL cell line - LR.

[0030] FIG. 23 is a heat map of the changes in gene expression in RC cell line following treatment with rimonabant. Detailed Description

[0031] In the following description, numerous specific details are set forth in order to provide a thorough understanding of the various embodiments. In other instances, well-known processes and methods may not be described in particular detail in order not to unnecessarily obscure the embodiments described here. Additionally, illustrations of embodiments here may omit certain features or details in order to not obscure the embodiments described here.

[0032] In the following, reference is made to the accompanying drawings that form a part of the specification. Other embodiments may be utilized, and logical changes may be made without departing from the scope of the disclosure. Therefore, the following detailed description is not to be taken in a limiting sense.

[0033] The description may use the phrases "in some embodiments," "in various embodiments," "in certain embodiment," or "in embodiments," which may each refer to one or more of the same or different embodiments. Furthermore, the terms "comprising," "including," "having," and the like, as used with respect to embodiments of the present disclosure, are synonymous.

[0034] Embodiments include theranostics compositions containing label-chelator-medical cannabinoid analog conjugates. The labels can be radionuclides that are used to label a medical cannabinoid analog through a chelator. Certain embodiments include cyclam (N4) as the chelator.

[0035] As used herein, the term "theranostic" refers to agents or applications that can function in both diagnostic and therapeutic modalities.

[0036] As used herein, the term "chelators" refer to compounds that form coordination complexes upon binding with metal ions or other substrates. The structure of chelating ligands and the metals that are chelated to them may be varied depending on the desired use. Many ligands that bind to radionuclide metals are tetradentate and contain a combination of four nitrogen and/ or sulfur metal coordinating atoms (i.e. N4, N3S, NZS2 and the like). Example of chelators that can be used here includes cyclam compounds (N4), diethylentriamine pentaacetic acid (DTPA), tetraaZacyclododecane-Ν,Ν',Ν", N"'-tetraacetic acid (DOTA), ethylenediaminetetraacetic acid (EDTA); dimercaptosuccinic acid (DMSA), sulfur colloid, and N2S2 systems such as MAMA (monoamidemonoaminedithiols), DADS (NZS diaminedithiols), CODADS and the like. These chelator systems and a variety of others are described in Liu and Edwards, Chem Rev. 1999, 99 (9), 2235-2268; N2S2 is also described in US. Pat. Nos. 4,897,225; 5,164,176; or 5,120,526. Method for synthesis of certain N4 compounds is described in U.S. Pat. No. 5,880,281 but they can also be obtained from commercial sources such as Sigma Aldrich Chemical (Milwaukee, Wis.) and, TCI America (Portland, OR). Certain N4 compounds which can be used as chelators may include but not limited to, 1,4,7, 10-tetraazacyclododecane (cyclen), 1,4,7,10- tetraazacyclotridecane (Cyclam 13), 1,4,7,11-tetraazacyclotetradecane (Isocyclam), 1,5,9,13- tetraazacyclohexadecane, 1,5,9,13-tetraazacycloheptaadecane, 1,5,9, 14-tetraazacyclooctadecane, 1,5,10,14-tetraazacyclooctadecane, 1,5,10,15-tetraazacyclodononadecane, are described in U.S. Pat. No. 8,758,723, US 2012/0276005, 6,093,382; 5,608,110; 5,665,329; 5,688, 487. PCT/GB2005/002807. Other examples of chelator moieties includes but not limited to, tetraazacyclododecane-N,N',N",N"'-tetra-acetic acid, monoamide (DOTA-MA); 10-(2- hydroxypropyl)- 1,4,7, 10-tetraaZacy clododecane-l,4,7-triacetic acid (HP-D03A). N4 is conjugated to a medical cannabinoid compound and further chelated to a metal. N4 has a closed- ring structure that helps stabilize the radionuclides. Chelators with higher lipophilicity, such as N4, also confer decreased renal and hepatic toxicity because they have shown decreased accumulation in these organs, resulting from greater uptake by the targeted cells. Conjugation of DOTA to highly selective CB?. receptor inverse agonist SR144528 following by chelation of Gallium (Ga), Technetium (Tc), Copper (Cu) or with lanthanide series such as Gadolinium (Gd), Europium (Er), Terbium (Tb) is described in U.S. Patent No. 8,367,714. Imaging CB] receptor using various radiotracers is described in PCT/US2009/043491. Radioligands with high affinity and selectivity for CBi receptors such as 3,4-diarylpyrazoline derivatives were labeled a radioisotope selected from the group consisting of ²H, ¹⁴C,ⁱ N, ⁱ⁸F, ¹⁵ Br, ^/6Br,^l23I for imaging with PET or SPECT. U.S. Patent Nos. US20050070596, 8,840,865, 9,617,215, 8,323,621 and WO2007130361 describe imaging of cannabinoid system for medical and therapeutic purposed to treat for instance inflammatory diseases, cancer, neurological disorders therapeutics and medical imaging.

[0037] As used herein, the term "label" refers to an atom, a molecule, or a compound that is used to identify the location of the composition to which the label is attached. Labels can have one or more of fluorescent, phosphorescent, luminescent, electroluminescent, chemiluminescent or other spectroscopic properties. These properties enable the detection and identification of the label- chelator-medical cannabinoid analog conjugates using any technique capable of detecting and identifying the label, including visible light, ultraviolet and infrared spectroscopy, Raman spectroscopy, nuclear magnetic resonance, positron emission tomography, and other methods known in the art.

[0038] As used herein, the term "imaging" refers to all tissue visualization processes using electromagnetic wave technologies for which the instant compositions can be used, including but not limited to cells of the nervous system, blood cells, cancerous cells, and precancerous cells. Provided here are kits for imaging neurologic cells. In an embodiment, the kit contains a predetermined quantity of a conjugate of a chelator and a medical cannabinoid analog; and a predetermined quantity of an imaging agent. The conjugate of the chelator and the medical cannabinoid analog can be present in the kit as precursors that subsequently interact with the imaging agent, when provided with suitable reaction conditions. The kit can also include a tin- containing reducing agent. Also provided here are kits for genomic or other omic assays that contain a predetermined quantity of a conjugate of a chelator and a medical cannabinoid analog; and a predetermined quantity of an imaging agent.

[0039] As used herein, the terms "radionuclide," "radioactive nuclide," "radioisotope," or "radioactive isotope" are synonymous. One or more different radioisotopes can be used as labels. The non-limiting examples of radionuclides include ^99mTc, ^117mSn, ¹⁷⁷Lu, ¹⁸⁸Re, ¹⁸⁶Re, ¹⁵³Sm, ¹⁶⁶Ho, ⁹⁰Y, ⁸⁹Sr, ⁶⁷Ga, ⁶⁸Ga, ^mIn, ¹⁸³Gd, ⁵⁹Fe, ²²⁵Ac, ²¹²Bi, ²¹¹At, ⁴⁵Ti, ⁶⁰Cu, ⁶¹Cu, ⁶⁷Cu, "Cu and ⁶²Cu. In other aspects, the metal ion is a non-radioactive metal such as ¹⁸⁷Re, ⁶⁹Ga, ¹⁹³Pt.

[0040] As used herein, the term "cannabinoid analog" refers to a compound capable of either interacting with cannabinoid receptors in a subject or sharing chemical similarity with cannabinoids or both. Cannabinoid analogs include synthetic or natural cannabinoid compounds that can function as agonists or antagonists. Embodiments of cannabinoid analogs include, but are not limited to, cannabidiol (CBD), cannabidiolic acid (CBDA), cannabidiol monomethylester (CBDM), cannabidiol-C4 (CBD-C4), cannabidivarinic acid (CBDA), cannabidivarin (CBV), cannabidiorcol (CBD-Ci), tetrahydrocannabinol (THC), N-arachidonoylethanolamine (AEA) or anandamide, 2-arachidonoylglycerol (2-AG), rimonabant, AM6538, taranabant, otenabant, cannabigerolic acid monomethylether (CBGAM), cannabigerol monomethylether (CBGM), cannabigerovarin (CBGV), cannabigerovarinic acid (CBGVA), cannabichromenic acid (CBCA), cannabichromene (CBC), cannabichromevarinic acid CBCVA), cannabichromevarin (CBCV), Δ⁹- tetrahydrocannabinolic acid A (THCA-A), A⁹-tetrahydrocannabinolic acid B (THCA-B), Δ⁹- tetrahydrocannabidiol (THC), A⁹-tetrahydrocannabinolic acid-C₄ (THC-C₄), Δ⁹- tetrahydrocannabivarinic acid (THCVA), A⁹-tetrahydrocannabivarin (THCV), Δ⁹- tetrahydrocannabiorcolic acid (THCA-Ci), A⁹-tetrahydrocannabiorcol (THC-Ci), A⁷-cis- isotetrahydrocannabivarin (THCV), A⁸-tetrahydrocannabinolic acid (A⁸-THCA), Δ⁸- tetrahydrocannabidiol (A⁸-THC), cannabicyclolic acid (CBLA), cannabicyclol (CBL), cannabicyclovarin (CBLV), cannabielsoic acid A (CBEA-A), cannabielsoic acid B (CBEA-A), cannabielsoin (CBEA-A), cannabinolic acid (CBNA), cannabinol (CBN), cannabinol methylether (CBNM), cannabinol-C₄ (CBN-C₄), cannabinol-Ci (CBN-C2), cannabiorcol (CBN-Ci), cannabinodiol (CBND), cannabinodivarin (CBVD), cannabitriol (CBT), lO-ethoxy-9-hydroxy- A^6a-tetrahydrocannabinol, 8,9-Dihydroxy-A^6a-tetrahydrocannabinol, cannabitriolvarin (CBTV), and ethoxy-cannabitriolvarin (CBTVE).

[0041] This disclosure provides for the generation and use of novel agents for precision medicine. Embodiments include compositions that provide a theranostics approach targeting the endocannabinoid system (ECS) implicated in the pathogenesis of neurologic disorders and cancer. This individualized medicine platform allows to deliver personalized medicine designed on the basis of individual genetic make-up, biochemistry, molecular imaging, molecular blueprint, and clinical observations and measurements associated to each patient's disease.

[0042] Embodiments described here include novel compositions and methods of using these compositions for imaging or treatment of diseased cells. Certain embodiments include compositions containing a cannabinoid analog and a chemotherapeutic agent. Certain embodiments include compositions containing a chemotherapeutic agent and a cannabinoid analog conjugated to a chelator. In certain embodiments, the chelator is cyclam. In certain embodiments, the cannabinoid analog is a cannabidiol. Certain embodiments include compositions containing a combination of a chemotherapeutic agent and a cannabinoid analog conjugated to a chelator and a label. In certain embodiments, the chemotherapeutic agent is Bruton's tyrosine kinase inhibitors, such as ibrutinib (IBN) and zanubrutinib (BGB). In certain embodiments, the chemotherapeutic agent is a proteasome inhibitor, such as carfilzomib (CFZ). In certain embodiments, the chemotherapeutic agent is tumorex (TMX).

[0043] Embodiments described here include novel compositions and methods of using these compositions for imaging diseased cells. These methods can be applied in the diagnosis, assessment, and treatment of any medical disorder with pharmaceuticals, medical cannabinoids, and combinations thereof. The diseases may include various forms of neurologic disorders and cancer. In particular, cancer can include one or more carcinoid, neuroendocrine cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, brain cancer, liver cancer, cervical cancer, colon cancer, renal cancer, skin cancer, head and neck cancer, bone cancer, esophageal cancer, bladder cancer, uterine cancer, lymphatic cancer, stomach cancer, pancreatic cancer, testicular cancer, colorectal cancer, and cancers of hematopoietic origin such as lymphoma, or leukemia. The neurologic diseases can include Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), multiple sclerosis, posttraumatic stress disorder (PTSD), epilepsy, seizures, Tourette's syndrome, schizophrenia, anxiety disorders, autism, depression, dementia, and other diseases and disorders that implicate the nervous system.

[0044] These methods have benefits to enhance existing imaging modalities that may include improvement in sensitivity and specificity, improvement of patient convenience, and reduction of adverse effects, time and costs. These methods for imaging diseased cells or a group of cells at a site of a diseased tissue or organ of a subject will enable health professionals to diagnose a subject, especially when that subject is being treated or under treatment with medical cannabinoids. Certain embodiments include methods of imaging at the site of a disease in a given subject to perform a pre- or post- treatment evaluation and to be able to monitor that subject for as long as that subject is being treated or under treatment with medical cannabinoids. In certain aspects, the method comprises detecting a signal generated by an imaging agent-labeled chelator-medical cannabinoid analog composition at the site of the disease of a subject, where the diseased cells, if present, generate a more intense signal than the cells in the surrounding tissue.

[0045] As used herein, the term "subject" refers to all kinds of animals including humans, rodents, other mammals, or avian species. The administration of the imaging agent-labeled chelator-medical cannabinoid analog conjugates can serve as a diagnostic agent, a prognostic agent, or an agent to alleviate or treat a disease in a subject. The target site can be any tissue of the subject, including but not limited to the brain, heart, lung, esophagus, intestine, breast, uterus, ovary, prostate, testis, stomach, bladder, or liver. Also, embodiments provided herein can be used as agents to target diseases, such as cancer or neurologic, gastrointestinal, metabolic, and neuroendocrine disorders. As used herein, the term "administration" refers to an activity of introducing a composition described herein to a subject by an appropriate method, and the composition may be administered via various routes of intravenous, oral, intramuscular, transdermal, intra-peritoneal, topical, sublingual, buccal, inhalation, nasal, or ophthalmic routes as long as they can deliver the same to the target tissues. The compositions described herein can be delivered as pharmaceutical formulation. A 'pharmaceutical formulation" refers to a mixture of one or more of the compounds described herein, or a pharmaceutically acceptable derivative as an active ingredient, and at least one pharmaceutically acceptable carrier or excipient. The purpose of a pharmaceutical composition is to facilitate administration of a compound to a subject. In another aspect, a pharmaceutical composition can contain a compound of one of the formulae described herein, or a pharmaceutically acceptable derivative, and a pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition includes two or more pharmaceutically acceptable salts, acids, esters, excipients, carriers, diluents, and combinations thereof.

[0046] The term 'pharmaceutically acceptable derivative" as used herein refers to and includes any pharmaceutically acceptable salt, pro-drug, metabolite, ester, ether, hydrate, polymorph, solvate, complex, and adduct of a compound described herein which, upon administration to a subject, is capable of providing (directly or indirectly) the active ingredient. For example, the term "a pharmaceutically acceptable derivative" of compounds described herein includes all derivatives of the compounds described herein (such as salts, pro-drugs, metabolites, esters, ethers, hydrates, polymorphs, solvates, complexes, and adducts) which, upon administration to a subject, are capable of providing (directly or indirectly) the compounds described herein.

[0047] As used herein, the term 'pharmaceutically acceptable salt" refers to those salts, which retain the biological effectiveness and properties of the parent compound. And unless otherwise indicated, a pharmaceutically acceptable salt includes salts of acidic or basic groups, which may be present in the compounds of the formulae disclosed herein. The present disclosure also provides certain processes, as examples, for the preparation of the above pharmaceutically acceptable salts, their derivatives, their analogs, their tautomeric forms, their stereoisomers, their polymorphs, and pharmaceutical compositions containing them.

[0048] Certain embodiments relate to pharmaceutically acceptable salts formed by the compounds described herein, their derivatives, their analogs, their tautomeric forms, their stereoisomers, their polymorphs and pharmaceutically acceptable compositions containing them. Typical inorganic acids used to form such salts include hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric, and the like. Salts derived from organic acids, such as aliphatic mono and dicarboxylic acids, phenylsubstituted alkanoic acids, hydroxyalkanoic and hydroxyalkandioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, may also be used. Such pharmaceutically acceptable salts thus include acetate, phenylacetate, trifluoroacetate, acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2- benzoate, bromide, isobutyrate, phenylbutyrate, beta-hydroxybutyrate, chloride, cinnamate, citrate, formate, fumarate, glycolate, heptanoate, lactate, maleate, hydroxymaleate, malonate, mesylate, nitrate, oxalate, phthalate, phosphate, monohydro genphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, propionate, phenylpropionate, salicylate, succinate, sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate, benzenesulfonate, p- bromophenylsulfonate, chlorobenzenesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate, naphthalene- 1- sulfonate, naphthalene-2- sulfonate, p-toluenesulfonate, xylenesulfonate, tartarate, and the like.

[0049] Certain embodiments include novel methods to synthesize agents that are conjugates of a chelator and a targeting ligand. Such agents may be used for imaging, diagnostic and/or therapeutic purposes. Accordingly, an embodiment includes a method of synthesizing a chelator- targeting ligand conjugate containing N4 and medical cannabinoid analogs. The method can further include synthesis with a metal in the form of an organometallic or a photon source. In some aspects, the metal ion is a radionuclide as described here.

[0050] Embodiments also include kits to prepare an imaging probe, a diagnostic agent, or a pharmaceutical composition. Certain specific embodiments include methods of imaging, diagnosing, or delivering a pharmaceutical composition to treat physiological disorders with medical cannabinoids. Accordingly, in an embodiment, any imaging modality can be used to detect signals from the one or more labels. Non-limiting examples of imaging methods used to detect the signals from the labels include PET, PET/CT, CT, SPECT, SPECT/CT, MRI, near-infrared (NIR), optical imaging, optoacoustic imaging, and ultrasound. Methods described here can be used to assess the personalized and efficacious dose and dosing regimens based on accurate evaluations determined through molecular imaging using medical cannabinoids.

[0051] Embodiments of the label-chelator-medical cannabinoid analog conjugate can include phytocannabinoids that are naturally occurring plant-derived cannabinoids. The active components in the medical marijuana strains Cannabis sativa and Cannabis indica are medical cannabinoids, which include more than 60 analog types, and two additional classes of bioactive molecules known as flavonoids and terpenoids, which are all naturally occurring compounds that are extracted and isolated from plants. Embodiments of the compositions that function as theranostics agents also include label-chelator-terpenoid analog conjugates, label-chelator-flavonoid conjugates, and label- chelator-phy to sterol conjugates. Among the natural medical phytocannabinoids, Δ⁹- tetrahydrocannabinol (A⁹-THC), cannabidiol (CBD), and cannabinol (CBN) are the most abundant, yet other phytocannabinoids can play a very important role in precision medicine. Based on structure, binding properties and signaling/function features, medical cannabinoids are grouped into distinct classes: (i) the classical medical cannabinoids, which consist of both natural plant extracts such as A⁹-THC and chemically synthesized compounds such as Marinol; (ii) Nonclassical medical cannabinoids, mainly exemplified by the synthetic cannabinoid receptor (CB) agonist CP- 55,940; (iii) Aminoalkylindoles, which consist of chemically produced cannabinoids like AM1241; (iv) Diarylopyrazoles, mainly comprise CB inverse agonists (or antagonists) such as SR141716A, also known as rimonabant; and (v) endogenous endocannabinoids, which are naturally produced by animal and human cells and include N-arachidonoylethanolamine, (AEA) or anandamide, 2-arachidonoylglycerol (2-AG), noladin ether, virodhamine and N- arachidonylodopamine (NAD A).

[0052] Embodiments of the label-chelator-medical cannabinoid analog conjugate can include the medical cannabinoids that have been approved for clinical research worldwide, as dronabinol, nabilone, nabiximols, cannador, cannabidiol, cannabinol, cannabigerol, tetrahydrocannabivarin, and cannabichromene. Embodiments of the label-chelator-medical cannabinoid analog conjugate can include medical cannabinoids such as HU-210, A⁹-THC, A⁸-THC and desacetyl-L-nantradol, which are recognized as CB1/CB2 receptor agonists, without distinctive specificity for either receptor. A⁹-THC stands out as a C. sativa cannabinoid, which exhibits CB 1/CB2 affinity and the highest psychotropic effects. By a pentyl substitution on A⁸-THC side chain, conversion into the HU-210 analog occurs, with increased receptor affinity. Other structural modifications of the THC backbone lead to new and selective CB₂ agonists JWH-133, JWH-139, and HU-308 and L-759633 and L-759656, which display affinities at the nanomolar range.

[0053] Embodiments of the label-chelator-medical cannabinoid analog conjugate can include non-classical medical cannabinoids, which are a family of bicyclic (AC) and tricyclic ACD medical cannabinoids. They are prominently represented by CP55940, along with CP55244 and CP47497 analogs. Of note, CP55940 is the best-known medical cannabinoid agonist, which displays a potent in vivo effect via shared CBi and CB₂ signaling.

[0054] Embodiments of the label-chelator-medical cannabinoid analog conjugate can include aminoalkylindoles. R-(+)-WIN55212 is the prototype of this family with medical cannabinoid-like features, which can bind both CBi and CB₂ receptors but exhibits higher specificity for CB₂ and can mimic in vivo THC-mediated effects. Other analogs like JWH-015 and L-768242 also show similar CB₂ affinity as R-(+)-WIN55212.

[0055] Embodiments of the label-chelator-medical cannabinoid analog conjugate can include medical diarylpyrazoles, which are a class of medical cannabinoid analogs whose distinctive function is to inhibit CBi or CB₂-dependent intracellular signaling pathways, acting as antagonists (inverse agonist); for example, embodiments can include SR141716A, AM251 and AM281 that inhibit CBi receptor mediated effects.

[0056] Embodiments of the label-chelator-medical cannabinoid analog conjugate can include certain endocannabinoids. N-arachidonoylethanolamine (AEA), or Anandamide, is an example of an eicosanoid that is converted into its active form, via the omega-3 (co-3) and omega-6 (co-6) biosynthetic fatty acid pathways, and specifically targets CB receptors in mammals. Consequently other eicosanoids that can function in these embodiments include methanandamide (R and S isomers), arachidonyl-2-chloroethylamide (ACE A), arachidonylcyclopropylamide (ACPA) and 2- arachidonoylglycerol (2- AG), which exhibit binding affinity to CBi and CB₂.

[0057] Embodiments of the label-chelator-medical cannabinoid analog conjugate can be used as theranostic agents targeting serious pathologies, such as cardiovascular, neurological, psychiatric, immunological, endocrine and neoplastic disorders. For instance, specific formulations of cannabinoid compounds are able to reduce proliferation and induce tumor cell death. Moreover, the mechanisms behind the anti-proliferative cell-killing effects (up to 70%) are known to result from intracellular signals driven by the binding of the cannabinoid agents to specific GPCRs, CBi and CB₂ of the ECS.

[0058] The ECS facilitates rapid local response to pathologic states or diseases. For example, when signals resulting from increased activation of neuronal Ca²⁺ channels or cellular stress cascades are transduced, immediate biosynthetic conversion of membrane phospholipids ensues, which leads to production and secretion of anandamide. Then, the endocannabinoid transmitters bind and activate target CB receptors, which in turn modulate adenylyl cyclase catalytic function to homeostasis and reduce cyclic adenosine monophosphate (cAMP) levels and protein kinase A activity, thus achieving equilibrium of Ca²⁺ fluxes and simultaneously correcting potassium (K⁺) channels. Upon completion of their regulatory function, Anandamide and or 2- AG undergo physiological degradation via the Fatty Acid Acyl Hydrolase (FAAH) activity or analog enzymatic pathways. Notably, the ECS also has the capacity to regulate the levels of its target CBi or CB₂ receptors, particularly in response to cell stress signaling. This function could be interpreted as a positive feedback mechanism to modulate the degree of signal transmission in certain pathologic states such as neuropathic pain and multiple sclerosis. The ECS system works conversely in disorders, such liver fibrosis or colorectal carcinoma. In the former, CBi levels are upregulated and thus signaling could result in deleterious progression toward cirrhosis of the liver, whereas in the latter, the low CBi expression will impinge upon analgesic benefits. On the other hand, endo and exo medical cannabinoids crosstalk with the opioid, serotonin, and N-methyl-d-aspartate (NMD A) nociceptive (pain) circuitry networks.

[0059] Embodiments of the label-chelator-medical cannabinoid analog conjugate can be used to evaluate the impact of medical cannabinoids in health and disease and to determine the early success or failure for specific applications. The label can be used to monitor the effect of the specific medical cannabinoid analog in the management of chronic pain. This can be achieved by imaging the medullary periaqueductal grey and rostral ventromedial areas to monitor analgesic effects. Additionally, one can take advantage of specific medical cannabinoid analogs targeting CB₂ that are selectively expressed by sensory neurons at the dorsal root ganglion, the spinal cord, and brain regions, to study and monitor the mechanisms of medical cannabinoid-driven analgesia.

[0060] Embodiments of the label-chelator-medical cannabinoid analog conjugate can be used to dissect and better understand the ECS and other pathways that modulate parallel functions aimed at protecting and maintaining a subject's homeostasis. CBi is mainly expressed in central and peripheral nervous system but discrete distribution has been observed in other tissues. Thus, utilization of the label-chelator-medical cannabinoid analog conjugates directed to CBi signaling can result in broad and pleiotropic actions. Although the CB i receptor has been known to primarily regulate functions associated to cognition, memory, perception, mood, behavior and psychotropic activities, there is increasing evidence that it can play a role in analgesia; cardiovascular, respiratory, and reproductive functions; as well in the maintenance of overall homeostasis. CB₂ receptors are predominantly expressed in immune competent organs where cells undergo antigen- dependent maturation and selection programs, which empowers them to survey and mount powerful responses against pathogens and aberrantly developed cells. CB₂ exhibits a relatively limited distribution in the central nervous system (CNS). CB₂-dependent activation involves regulatory mechanisms that support immune cell migration to the site of inflammation and the release cytokines. The expression of CB₂ is particularly important for CNS microglia, as demonstrated by the capacity of medical cannabinoid agents to reduce cytokine-mediated neuro- inflammation. Embodiments of the label-chelator-medical cannabinoid analog conjugate that include specific CB₂ ligands, such as 0-3223 (a synthetic CB₂ specific agonist), can be used for anti-inflammatory and anti-nociceptive applications, without apparent CBi-like mediated effects.

[0061] This disclosure also provides for innovative radiopharmaceutical imaging analogs that can be used to diagnose, stage, restage and treat central nervous, cardiovascular and cancer diseases. Imaging with radiolabeled physiological agents can be implemented using nuclear molecular agents, which provide accurate measurements of molecular pathways, by integrating real-time nuclear imaging protocols. This approach is designed to enhance clinical capability for patient selection, improve pharmacokinetic assessments, optimize dosage formulations and predict therapy outcomes. The overall impact of the comprehensive application of theranostics (diagnosis/therapy/prognosis) technology to medical cannabis platforms, stems from the evolving versatility of nuclear imaging analogues and the emerging competence of hybrid instrumentations, which will provide precision, sensitivity, specificity and real-time data tailored to individual patient's diagnosis and therapy.

[0062] Compositions containing the label-chelator-terpenoid conjugates can be used to study, monitor, and provide the potent antioxidant, anti-inflammatory, analgesic, anticancer, antibiotic and anti-psychiatric (anxiety and depression) benefits of terpenoid compounds.

[0063] Compositions containing the label-chelator-flavonoid conjugates can be used to study, monitor, and provide the antioxidant, anti-inflammatory and anticancer properties of flavonoid compounds, including polyphenol cannabis flavonoids. The cannabis flavonoids can provide significant cardiovascular protection, particularly improving coronary and peripheral circulation by maintenance of homeostatic blood pressure, prevention of the formation of blood clots, and reduction of atherosclerosis risks. Mechanisms of cannabis flavonoids mediated antioxidative and anti-inflammatory effects include apigenin (4',5,7-trihydroxyflavone)-dependent inhibition of TNF-α, which has also shown to exhibit therapeutic benefits in multiple sclerosis and rheumatoid arthritis.

[0064] Compositions containing the label-chelator-phytosterol conjugates can be used to study, monitor, and provide the cardiovascular protection, anti-inflammation, and anti-systemic edema properties of phytosterol compounds, including medical cannabis phytosterols.

[0065] Embodiments of the label-chelator-medical cannabinoid conjugate can utilize tetra- azacyclic (N4) conjugation-based technology, also known as N4-technology, which can also be adapted for serial multi-imaging/therapy of a subject. Here, the hydrophobic chelator(s)-based N4- technology coordinates metals to form a complex that may be conjugated to hydrophobic/hydrophilic molecules, including medical cannabinoid analogs to produce novel compounds or to better understand the bio-distribution scope and targeting capabilities of exiting compounds. N4 conjugation of novel or existing compounds can be used for purposes including imaging, cold therapy, and radiotherapy that targets specific cell receptors, pathways, tissues or organs, using medical cannabinoids. Moreover, certain N4 compounds may be obtained from other commercial sources, which are noted in U.S. Pat. No. 8,758,723. In addition, metal isotopes are either obtained from generators such as ^99mTc, ¹⁸⁸Re and ⁶⁸Ga, or from commercial sources such as ⁶⁴Cu and ^luIn, which are available and cost-effective. N4-conjugates provide a simple platform for small molecules, peptides, or antibodies with high purity and efficient formulation to target components of the endocannabinoid system. Moreover, radiolabeled N4-conjugates provide molecular target assessment in the areas of abnormal cell cycle and epigenetic microenvironments. Similarly, N-4 theranostic applications offer post-treatment evaluation and can be tailored for internal radio-therapeutics, when the conjugate is labeled with ¹⁸⁸Re or ¹⁷⁷Lu (beta and gamma emitters) or ²²⁵ Ac, ²²³Ra (alpha emitters) for simultaneous SEE and TREAT approaches. Moreover, the N4 technology provides the flexibility to prepare conjugates with or without cold platinum metals, thus enabling the N4 chemistry to increase the cold payload of medical cannabinoids to optimal concentrations for cellular and immunotherapy as theranostics agents. The radiolabeled N4-conjugates are focused on the imaging of immuno-modulatory functions and the therapy of oncological and neurological diseases, via cannabinoid receptor/transporter-mediated mechanisms. This medical cannabinoid precision-based technology platform integrates diagnostic imaging (molecular imaging) with innovative tools to understand the dynamic changes in pathway- activated cell receptors leading to tissue degeneration, inflammatory, and proliferative disorders to improve patient diagnosis, therapy and prognosis. This technology allows for the medical cannabinoids to be utilized as a therapeutic for treatments amenable to cannabinoids alone or as combinations with radiopharmaceuticals, immunopharmaceuticals, and other compounds.

[0066] Several synthetic antagonists taranabant, otenabant, and AM6538, as well as the inverse agonist/antagonist rimonabant show great promises as therapeutic agents. For example, in targeting obesity, rimonabant showed to promote weight loss, weight maintenance and to prevent weight regain in patients with a BMI of greater than 27 kg/m², when accompanied with at least one risk factor such as hypertension or type 2 diabetes. However, rimonabant had increased frequencies of psychiatric adverse events, including major depression, dysthymia, seizures and suicide. These adverse effects discouraged its development as an approved therapeutic intervention for other disease states. While the cause of the adverse events has been speculated to result from the inverse agonist mechanism of the CB i receptor activity, many factors have not been considered where this technology platform could resolve other medical conditions. Likewise, agonists like the natural medical cannabinoid THC, have demonstrated great promise for therapeutic intervention. For example, medical THC has demonstrated benefits with refractory nausea and vomiting for patients undergoing chemotherapy along with alleviating neuropathic pain and spasticity. However, medical THC is psychoactive, which can also lead to lethargy, postural hypotension and decreased motor coordination when overdosing occurs. Conversely, given to the right patient, disease, and dose, it is conceivable that the effects against a disease by pathology biomarker- guided N4-THC theranostic imaging, can actually result in favorable clinical outcomes.

[0067] Combinations containing the label-chelator-medical cannabinoid analog conjugates combine imaging with the therapeutic intervention and can image in real time the uptake and activity of N4 conjugated medical cannabinoids, which is essential to select the individual patient with the targeted dysfunctional pathway (right disease) and to assess optimal dosage (right dose). This approach allows for visually seeing the composition located at the tissue site and determining the actual dose of uptake to that site for that patient. This platform allows one to evaluate: (a) if dosing is the cause of the adverse event; (b) if bioavailability is the cause or (c) if there is a limited uptake and/or bio-distribution. In keeping with these parameters, the embodiments serve to dissect effects that are patient dependent, particularly if one of these effects are genetic, epigenetic, or exhibit allelic variations associated to the individual's ECS. [0068] The effective amount of a compound is determined based on several factors, such as age and weight of the patient, severity of the disease, other co-existing factors. The effective amount of a compound includes exemplary dosage amounts for an adult human of from about 0.1 to 100 mg/kg of body weight of active compound per day, which may be administered in a single dose or in the form of individual divided doses, such as from 1 to 4 times per day. It will be understood that the specific dose level and frequency of dosage for any particular subject may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the species, age, body weight, general health, sex and diet of the subject, the mode and time of administration, rate of excretion, drug combination, and severity of the particular condition.

[0069] Furthermore, some medical cannabinoids have not yet been explored and this platform lays a foundation for conjugating several medical cannabinoids for investigating the effects in diverse physiological disorders, through a precision medicine approach. The N4 technology can allow for determination of the effectiveness in targeting, uptake and distribution; wherein the conjugates are imaged further allowing for the assessment of efficacy. Embodiments include the pharmaceutical formulation, kit preparation, and method of use and administration of pathway- directed molecular agents for non-metal/metal labeling and metallic/cold therapeutics, when such agents are prepared in aqueous or organic conditions. Synthesis and purification in organic solvents requires the use of protecting and de-protecting chemicals to achieve desired compositions.

[0070] Embodiments also include kits containing components to prepare an imaging probe that has a conjugate of a label, a chelator, and a cannabinoid analog. Embodiments also include kits containing components to prepare a diagnostic agent based on a conjugate of a label, a chelator, and a cannabinoid analog. Kits can also be constructed to contain components to deliver effective amounts of pharmaceutical compositions containing a conjugate of a label, a chelator, and a cannabinoid analog. Certain embodiments include kits for diagnostic imaging of a site of neurologic disorders and cancers, and delivery of a dual therapeutic intervention agent to a subject. The conjugate of the chelator and the cannabinoid analog can be present in the kit as precursors that subsequently interact with the imaging agent, when provided with suitable reaction conditions. For example, a kit can include one or more sealed containers contain a predetermined quantity of a chelator-medical cannabinoid analog conjugate and one or more sealed containers that contain a predetermined quantity of a second imaging agent that can be applied in imaging the brain of a subject. In some embodiments, the kit further includes a sealed container containing a radionuclide. In certain embodiments, a kit further includes a predetermined quantity of a chelator- medical cannabinoid analog conjugate and a sufficient amount of a reducing agent to label the conjugate compound with a radionuclide metal ion. In certain embodiments, a kit further includes a predetermined quantity of a chelator-medical cannabinoid analog conjugate and a reducing agent along with a buffer solution, wherein the reducing agent includes, but not limited to, tin(II) chloride. The kit may also contain other components, such as pharmaceutically acceptable salts, buffers, antioxidants, and preservatives. In certain embodiments, buffer solutions are phosphate buffer solutions to stabilize the chelator-medical cannabinoid analog conjugates. Phosphate buffer solutions may contain monosodium phosphate, disodium phosphate, or combination thereof, dissolved in water. In certain embodiments, an antioxidant is included in the chelator-medical cannabinoid analog conjugate composition to prevent oxidation of the chelator moiety. In certain embodiments, the antioxidant is vitamin C (ascorbic acid). Other antioxidants that can be used to include tocopherol, pyridoxine, thiamine, butylated hydroxyl toluene, sodium edetate, or rutin. In certain embodiments, a stabilizer is included in the chelator-medical cannabinoid analog conjugate composition to prevent degradation and to enhance shelf life or storage life of the chelator moiety. In certain embodiments, the stabilizer is mannitol. However, other components, such as sugars or bulking agents, may also be used, wherein the sugars are simple sugars, complex chain sugars, sugar alcohols or salts thereof. Stabilizers may include but are not limited to glucose, lactose, maltose, xylose, sorbitol, cellulose, or carboxymethylcellulose sodium. In certain embodiments the predetermined quantity of a chelator-medical cannabinoid analog conjugate may be present in dosing amounts to treat a neurologic disorder or cancer as a therapeutic intervention. The kit may contain the components in liquid, frozen, dry form, or lyophilized form.

Examples

[0071] Example 1. Synthesis of 1,4,8,1 l-Tetraazacyclotetradecane-l'-propyl-[N-(Piperidin-l- yl)-5-(4-chlorophenyl)-l-(2,4-dichlorophenyl)-4-methyl-lH-pyrazole-3-carboxamide] (also known as VYR207)

VYR207

[Formula I]

[0072] VYR207 can be synthesized in several ways. Shown below is Scheme 1 for the synthesis of l,4,8-Tris trifluoroacetyl)-l 4,8,l l-tetraazacyclotetradecane (N4-TFA3; TFA3-cyclam).

TFA3-cyclam

[Scheme 1]

[0073] Cyclam (N4;l,4,8,l l-tetraazacyclotetradecane; N4; 7.53 g, 37.58 mmol) was dissolved in 30 mL of methanol. To this clear solution, NEt₃ (5.20 mL, 37.58 mmol) was added in one portion. Then, ethyl trifluoroacetate (18.0 mL, 150.3 mmol) was added dropwise over a period of 5 minutes while stirring. The homogeneous reaction mixture was cooled with ice-water bath to control the mild exothermic reaction and stirred at 25 °C under the atmosphere of N₂ for 5 h. Volatiles were then removed under vacuum. The residue was dissolved in minimum amount of CH₂C1₂ (2.0 mL) and passed through a short silica gel pad (25 g), eluted with 100% EtOAc. The eluent was concentrated to give the product a white semi solid form (17.1 g, 92.5%). The resulting compound was analyzed by proton nuclear magnetic resonance (^lH NMR), carbon- 13 nuclear magnetic resonance (¹³C NMR), and high-resolution mass spectrometry (HRMS) by electrospray ionization (ESI). The Ή NMR data, ¹³C NMR data, and the HRMS-ESI determination of mass of VYR207 are as follows. [0074] The ^lH NMR data (200 MHz, CDC1₃): δ 3.85 - 3.25 (multiplet, 12 H), 2.80 (broad singlet, 2 H), 2.74-2.50 (broad singlet, 2H), 2.30- 1.90 (multiplet, 2 H), 1.85-1.63 (multiplet, 2 H), 1.25- 0.60 (multiplet, 1 H), and the ¹³C NMR data (75.5 MHz, CDCI3): δ 158.74- 157.31 (multiplet, C=0, multiplets due to existence of conformers), 122.84- 11.32 (quartet, CF3, due to C-F coupling, Jc-F ~ 264 Hz, further split due to existence of conformers), 51.2 - 46.2 (multiplet, CH₂ next to N), 29.4 - 27.8 (multiplet, CH₂); The HRMS-ESI mass calculated for Ci₆H₂iF₉N₄03 was C 39.35; H 4.33; N 11.47; 0 9.83; and the mass found was: C 39.19; H 4.36; N 11.33; O 10.04.

[0075] Shown below is Scheme 2: Synthesis of Bromopropyl-[N-(Piperidin- l-yl)-5-(4- chlorophenyl)-l-(2,4-dichlorophenyl)-4-methyl- lH-pyrazole-3-carboxamide] (Step 1 in Scheme

[Scheme 2]

[0076] N-(Piperidin-l-yl)-5-(4-chlorophenyl)- l-(2,4-dichlorophenyl)-4-methyl-lH-pyrazole- 3-carboxamide (SR141716, rimonabant; 3.70 g, 7.57 mmol) was added to a magnetically stirred anhydrous CH3CN (30 mL). The mixture was stirred at room temperature until a solution was obtained (~ 10 min). To this solution, K₂C03 (1.57 g, 11.43 mmol) and 1, 3-dibromopropane (2.29 g, 11.35 mmol) were then added. The mixture was refluxed under the atmosphere of N₂ for 16 h. The progress of the reaction was monitored by TLC (1: 1 EtOAc/Hexane). The mixture was cooled to room temperature and filtered through a sintered glass filter to remove insoluble salt (washed with 20 mL CH3CN). The solvent was then concentrated to give yellow oil (6.3 g, 70%). Structure of starting material SR141716 was determined by NMR (FIG. 1) and HPLC (FIG. 2). As shown in the Table 1, the peaks and retention times show elution of SR141716 at peak 6 (7.994 minutes) with significant purity, as evidenced by the area% being greater that 99% for that data point.

[0077] Table 1

[0078] Conditions for HPLC were as set forth below. An Athena C18 column with particle size of 3 micrometer (μιη) and physical dimensions of 2.1 millimeter (mm) diameter and 100 mm length. The two solvents used were 0.1 % phosphoric acid in 100% acetonitrile (Solvent A) and 0.1 % phosphoric acid in 100% acetonitrile (Solvent B). The two solvents were utilized to create a gradient as shown in Table 2. The HPLC system was operated at a flowrate of 0.5 milliliters per minute (mL/min) and detection at 210 nanometers (nm).

[0079] Table 2

[0080] This compound SR141716 was then subject to conjugation and deprotection to yield 1,4,8,1 l-Tetraazacyclotetradecane-l'-propyl-[N-(Piperidin-l-yl)-5-(4-chlorophenyl)-l-(2,4- dichloro phenyl) - 4-methyl-lH-pyrazole-3-carboxamide] (VYR207) (Steps 2-3 in Scheme 2). [0081] A mixture of bromopropyl-[N-(piperidin-l-yl)-5-(4-chlorophenyl)-l-(2,4- dichlorophenyl)-4-methyl-lH-pyrazole-3-carboxamide] (4.4 g, 7.57 mmol) in anhydrous CH3CN (20 mL) was magnetically stirred at room temperature. To the reaction mixture, KOH powder (g, 11.35 mmol) and TFA3-cyclam (11.35 mmol) were added and the mixture was stirred at room temperature for 1 hour. The solvent was evaporated on a rotary evaporator under reduced pressure. The residue was purified by flash chromatography using Hexane:EtOAc (6:4) as an eluent to afford TFA3-cyclam-conjugate as a white solid (16 g, 45 %). To a solution of the TFA3-cyclam- conjugate (3.0 g, 3.05 mmol) in MeOH (6.0 mL) K2CO3 (1.26 g, 9.1 mmol) was added in one portion. The suspension was heated under reflux for 3 h. Toluene (30 mL) was then added to the cooled mixture. MeOH was removed by forming an azeotrope with toluene. After MeOH solvent was completely removed, the hot toluene suspension with inorganic salt was filtered and concentrated to give a desired free base (6.2 g, 86 %) as a white solid.

[0082] Example 2 - Synthesis of 1,4,8,1 l-tetraazacyclotetradecane-l'-acetyl-[N-(Piperidin- yl)-5-(4-chlorophenyl)-l-(2,4-dichlorophenyl)-4-methyl-lH-pyrazole-3-carboxamide]

(VYR206).

[0083] The structure of target compound VYR206 (aliphatic chain with carbonyl group) shown as Formula II and complete synthesis of VYR206 is shown in Scheme 3.

[Formula II]

VYR206 can be synthesized in several ways. Shown below is a representative scheme,

[Scheme 3]

[0085] Step 1 in Scheme 3: Synthesis of l '-Ethylacetyl-[N-(Piperidiny-l-yl)-5-(4- chlorophenyl)- 1 -(2,4-dichlorophenyl)-4-methyl- lH-pyrazole-3-carboxamide]

[0086] N-(Piperidiny- l-yl)-5-(4-chlorophenyl)- l-(2,4-dichlorophenyl)-4-methyl-lH-pyrazole- 3-carboxamide; SR141716; 200 mg, 0.43 mmol) was added to a magnetically stirred anhydrous CH3CN (10 mL). The mixture was stirred at room temperature until a solution was obtained (10 min). To this solution, K2CO3 (200 mg, 1.44 mmol) and bromo ethylacetate (400 mg, 2.4 mmol) were then added. The mixture was refluxed under the atmosphere of N2 for 48 h. TLC (CHiCh/MeOH, 100:7) was used to monitor the reaction progress, which was completed after 4 - 8 hours. The mixture was cooled to room temperature and filtered through a sintered glass filter to remove insoluble salt (washed twice with 20 mL CH3CN). The residue was then isolated by column chromatography and eluted with methylene chloride followed by methylene chloride/methanol (100:6) to yield the desire compound as a white powder (428 mg, 55 %).

[0087] Step 2 in Scheme 3: Synthesis of l '-Acetic acid-[N-(Piperidin-l-yl)-5-(4- chlorophenyl)- 1 -(2,4-dichlorophenyl)-4-methyl- lH-pyrazole-3-carboxamide]

[0088] The deesterification of ethylacetyled compound from step 1 was carried out as follows: l '-ethylacetyl-[N-(Piperidin-l-yl)-5-(4-chlorophenyl)- l-(2,4-dichlorophenyl)-4-methyl- lH- pyrazole-3-carboxamide (200 mg, 0.36 mmol) was dissolved in dioxane (9 mL). To the reaction mixture, NaOH (250 μί, 10 N) was added along with 3 mL of water. The reaction mixture was heated at 50 °C for 4 h. The dioxane in the reaction mixture was evaporated and then water was added. HC1 (5 N) was slowly added to the mixture to reach pH at 4-5. Reaction mixture was then extracted with CH2CI2 to afford the product as a white solid (197 mg, 95%).

[0089] Steps 3-4 in Scheme 3: Conjugation and Deprotection of 1,4,8,11- Tetraazacyclotetradecane-1 '-acetyl- [N-(Piperidin-l-yl)-5-(4-chlorophenyl)-l-(2,4-dichloro phenyl)-4-methyl- lH-pyrazole-3-carboxamide] (VYR206):

[0090] To a solution of l'-Acetic acid-[N-(Piperidiny-l-yl)-5-(4-chlorophenyl)-l-(2,4- dichlorophenyl)-4-methyl-lH-pyrazole-3-carboxamide] (170 mg, 0.32 mmol) in anhydrous CH2CI2 (25 mL), l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (91 mg, 0.48 mmol), hydroxybenzotriazole (HOBt) (65 mg, 0.45 mmol), diethylamine (DEA) (300 μί) and 1,4,8-Tri- Boc-l,4,8,l l-tetraazacyclotetradecane (BOC3-cyclam; 163 mg, 0.32 mmol) were added. The mixture was stirred at room temperature for 48 h. The protected BOC3-cyclam-conjugate was isolated by flash column chromatograph using gradient CfhCh/MeOH (100: 1 to 100: 10) to yield (300 mg, 95 %). To de-protect the cyclam conjugate, the BOC3 -protected conjugate (175 mg, 0.17 mmol) was dissolved in TFA/CH2CI2 (8 mL; 2: 1). The reaction mixture was stirred at room temperature for 6 h. The solvent was then concentrated and the desired product was purified by flash chromatography eluted with Cf Ck/MeOH (100: 1 to 100:25) to afford the final compound as a white powder (150 mg, 81 %). The structure of VYR206 was determined by NMR, MS, and HPLC techniques (FIGS. 3, 4, and 5 respectively).

[0091] Example 3

N4-Cannabidiol (N4-CBD)

VYC203 [Formula III] [0092] According to the similar N4-strategy approach as described for VYR207 compound in Example 1, two compositions based on natural cannabinoid compounds N4-Cannabidiol (VYC203) and N4-Tetrahydrocannabidiol (VYT205) were synthesized.

^-Tetrahydrocannabinol (N4-THC)

VYT205 [Formula IV]

[0093] VYR203 and VYR205 can be synthesized in several ways. Shown below is a representative scheme, Scheme 4.

Cannabidiol (CBD) Tetrahydrocannabinol (THC)

N4-Cannabidiol (N4-CBD) N4-Tetrahydrocannabinol (N4-THC)

VYC203 VYT205

[Scheme 4] [0094] Briefly, Tetrahydrocannabinol (THC) (350 mg, 1.11 mmol) was added to a magnetically stirred anhydrous CH3CN (12 mL). The mixture was stirred at room temperature until a solution was obtained (~10 min). To this solution, bromopropyl-[l, 4, 8-Tris(trifluoroacetyl)- 1,4,8,11- tetraazacyclotetradecane] (bromopropyl-N4-TFA3 ; bromopropyl-TFA3; 280 mg, 0.3 mmol) and KOH (157 mg, 2.8 mmol) were then added. The mixture was stirred at room temperature for 1 h. The progress of the reaction was monitored by TLC (3:1 EtOAc/Hexane). The residue was purified by flash chromatography using Hexane:EtOAc (6:4) as an eluent to afford TFA3-cyclam-THC as a white solid (510 mg, 51 %). To a solution of TFA3-cyclam-THC conjugate in MeOH (4.0 mL) K2CO3 (420 mg, 7.2 mmol) was added in one portion. Suspension was heated under reflux for 3 h. Toluene (20 mL) was then added to the cooled mixture. MeOH was removed by forming an azeotrope with toluene. After MeOH solvent was completely removed, the hot toluene suspension with inorganic salt was filtered and concentrated to give a desired free base (575 mg, 76 %) as a white solid VYT205. The same synthetic route is applied to VYC203 compound.

[0095] Example 4

[0096] Immunoblot analysis of cell lysates from various DLBCL cell lines was performed to evaluate expression of the CBi receptor (FIG. 6A) with expression of β-actin as the control for sample loading (FIG. 6B), with each cell line identified by respective initials. Western blotting showed very high protein expression levels of CB 1 in the RC cell line in comparison to Tole, TMD, EJ, LP, LY3, and CJ cell lines, while no expression was detectable in FN, HF, DOHH2, and DBR cell lines.

[0097] Cell lysates from 24 DLBCL cell lines were analyzed for relative expression of CB 1 and CB₂ mRNAs. In certain cell lines, such as TJ, LY-19, CJ, and BJAB, CBi mRNA was expressed about 10-15 % more than CB₂ mRNA. In certain cell lines, such as SF, MZ, Toledo, DBr, SUDHL- 4, and MCA, CBi mRNA was expressed about 100-200 % more than CB₂ mRNA. In certain cell lines, such as SF, MZ, Toledo, DBr, SUDHL-4, and MCA, CBi mRNA was expressed about 100- 200 % more than CB₂ mRNA; and in cell lines, such as U2392 and EJ, CBi mRNA was expressed about 300-600 % more than CB₂ mRNA. In certain cell lines, such as LY-3 and Pheiffer, CB₂ mRNA was expressed about 2-10 % more than CBi mRNA. These results show CBi is a viable target in malignant immune cells, along with CB₂ that is known to be primarily associated. Furthermore, CBi may play a pivotal role in lymphoblastic cells, including determination of therapeutic sensitivity or resistance.

[0098] Example 5

[0099] Cell viability was assessed using MTS assays, a colorimetric method for the sensitive quantification of viable cells based on the reduction of the MTS tetrazolium compound to a colored formazan dye by NAD(P)H-dependent dehydrogenase enzymes in metabolically active cells. The formazan dye is quantified by measuring the absorbance at 490-500 nm. Cell viability assays (MTS) were performed to determine the viability of four DLBCL cancer cell lines— EJ, U2932, CJ and LY19 in vitro (72 h incubation) in the presence of four CB1/CB2 cannabinoid receptor ligands: CBi-selective antagonist CP945,598 (Otenabant), CBi-selective inverse agonists— SR141716 (rimonabant) and AM251, and CB₂-specific agonist AM1241. The effect of cannabinoid receptor ligand SR141716 on the viability of the DLBCL cancer cell lines— EJ, U2932, CJ and LY19 is shown in FIG. 7. The effect of cannabinoid receptor ligand CP945,598 (Otenabant) on the viability of the DLBCL cancer cell lines— EJ, U2932, CJ and LY19 is shown in FIG. 8. The effect of cannabinoid receptor ligand AM 1241 on the viability of the DLBCL cancer cell lines— EJ, U2932, CJ and LY19 is shown in FIG. 9. The effect of cannabinoid receptor ligand AM251 on the viability of the DLBCL cancer cell lines— EJ, U2932, CJ and LY19 is shown in FIG. 10. All compounds depicted in FIGS. 7-10 correspond to non-conjugated (N4 chelator-free) agents. As expected, CB₂ agonist AM 1241 demonstrated a significant effect, however CBi- selective antagonist and inverse agonists (CP945,598, SR141716 and AM251) also had significant results depending on the specific cell line. For example, SR141716 and AM251 appear to have a more potent effect on LY19 where AM251 appears to have the greatest potency for U2932. Overall, the compounds demonstrate that CBi as a target for therapeutic intervention. CB₂ is previously known to be associated with immune cells.

[00100] Example 6

[00101] In Examples 6-8, N4-conjugated compounds were analyzed. The effects of CBi inverse agonist SR141716 and its conjugated form-VYR206 were analyzed using diffuse large B-cell lymphoma (DLBCL; 16 cancer cell lines; FIGS. 11A and 11B) and mantle cell lymphoma (MCL; 8 cell lines; FIGS. 11C and 11D) cancer cells. Exposure of SR141716 to DLBCL (FIG. 11A) markedly decreased viability than VYR206 (FIG. 11B). Diffuse large B-cell lymphoma cancer cells were resistant to lower VYR206 concentration (< 50 μΜ) (FIG. 11B). The lower-dose of SR141716 (10-40 μΜ) drug compared to VYR206 indicated the reduction of cell viability (relatively steep initial slopes especially in Rec-1 and SP-53) in MCL cell lines (FIG. 11C). The results showed that exposure of MCL cancer cells to VYR206 (FIG. 11D) led to a substantial decrease in cell viability when compared with the all DLBCL cell lines (FIG. 11B). Exposure to 50 μΜ VYR206 drug had a significant effect on the induction of cell death (90 %) in MCL cell line Jeko and Mino (FIG. 11D). The less effective VYR206 may prove to be beneficial for controlling the dosing. Rimonabant by itself has severe adverse effects in patients with high or chronic dosing. Curtailing or modifying the potency may help to reduce adverse effects while still providing a therapeutic effect against cancerous cells.

[00102] Example 7

[00103] The effects of SR141716 and VYR206 were analyzed. MTS assays for cell viability (72 h incubation) were performed on diffuse large B-cell lymphoma (DLBCL) cells— LR, MS, RC and DOHH2 (FIGS. 12A - 12D) and mantle cell lymphoma (MCL) cells— Jeko, Mino, Rec-1 and JMP1 (FIGS. 13A - 13D). Preliminary data shows DLBCL cancer cell lines MS, DOHH2 and mantle cell lymphoma cell line REC-1 are more resistant to drug VYR206 with comparison to other cell lines from FIGS. 12B, 12C, and 13B. The results show that exposure of DLBCL to SR141716 led to induction of cell death (< 90 %) at drug concentration 50 μΜ in LR, MS and RC cell lines (FIGS. 12A, 12B, 12D; blue lines) with comparison to DOHH2 cell line, inhibition was > 50 % SR141716 was more effective to DLBCL than VYR206. Complete DLBCL inhibition with VYR206 was reached at drug concentrations: 200 μΜ for LR, MS and DOHH2 (FIGS. 12A, 12B, 12C) and 100 μΜ for RC (FIG. 12D). Mantle cell lymphoma cell lines JEKO, JMP-1 and Mino (FIGS. 13A, 13B, 13D, red lines) were less resistant to VYR206 than DLBCL. It was also observed from FIGS. 12 and 13 that N4 (cyclam) has been proven to have no effect on cell viability in all tested lymphoma cell lines.

[00104] Example 8

[00105] High throughput screening (THS) assessment of cell viability in 16 DLBCL (FIGS. 14 A and 14B) and 8 MCL (FIG. 14C) cell lines were assessed following treatment a 72-hour incubation of the cell lines with cannabidiol, which was obtained as CBD in a coconut oil extract. Cell viability, 90% of the all cells tested, remained viable in a concentration range of 0-5 μΜ CBD. Complete cell death was obtained at 25 μΜ CBD concentration only for two cancer cell lines, HT and EJ (FIG. 14B); 50 μΜ CBD concentration gave a complete cell death viability in seven cell lines MZ, HF, HB, CJ, MS, Toledo and EJ (FIGS. 14 A and 14B). Cancer cell line DOHH2 was less susceptible to lower CBD concentration (0-25 μΜ) (FIG. 14B). After exposure to CBD concentration range 15-20 μΜ 50 % cell viability was reached in 8 cell lines (MZ, HB, CJ, LP, MCA, MT, EJ) (FIGS. 14A and 14B). All screened mantle cell lymphoma cancer cells (Mino, Jeko, SP53, Maver-lZ138, JMP-1, Rec-1 and PF-1) were less sensitive at CBD concentration range from 15 μΜ to 25 μΜ (FIG. 14C).

[00106] Example 9

[Scheme 5]

[00107] ⁶⁸Ga-N4-Compound labeling process (Route A of Schemes 5, 6, and 7) was generally initiated by the addition of ⁶⁸Ga chloride (20 mCi in 0.1 N HC1) in vials. The reaction was heated at 70 °C for 15 min and the pH of the product was adjusted to 5-6 with sodium bicarbonate. The tracer product was validated using two analytical tools (reverse phase HPLC and radio-TLC) to determine the active pharmaceutical ingredient (API) purity, API yield, radiochemical purity and yield and specific ⁶⁸Ga-N4-compound activity. In certain compounds, there was a need to use C- 18 column to remove unreacted ⁶⁸Ga chloride or ⁶⁸Ga oxide, followed by elution with ethanol. ^99mTc-N4-compound labeling process (Route B of Schemes 5, 6, and 7) was generally initiated by the addition of tin (II) chloride (O.lg) and ^99mTc pertechnetate (20 mCi in 0.1 mL saline) in vials. The reaction was immediately chelated and the pH of the product was adjusted to 5-6 with saline. The tracer product was validated using two analytical tools (reverse phase HPLC and radio-TLC) to determine the active pharmaceutical ingredient (API) purity, API yield, radiochemical purity and yield and specific ^99mTc-N4-compound activity. Radiochemical purity of ^99mTc-N4- compounds was assessed by high-performance liquid chromatography (HPLC), equipped with Nal and UV detector (274 nm), and was performed using a C-18 reverse column with a mobile phase of acetonitrile:water (7:3) at a flow rate of 0.5 mL/min.

[00108] Radiochemical purity was also determined by radio-TLC (ITLC SG, Gelman Sciences, Ann Arbor, MI) eluted with ammonium acetate (1M in water): methanol (6: 1). High-performance liquid chromatography (HPLC), equipped with a Nal detector and UV detector (254 nm), was performed on a gel permeation column (Biosep SEC-S3000, 7.8 300 mm, Phenomenex, Torrance, CA) using a flow rate of 1.0 mL/min. The eluant was 0.1% LiBr in phosphate buffered saline (PBS) (10 mM, pH 7.4).

^-Tetrahydrocannabinol (N4-THC)

VYT205

[Scheme 7]

[00109] Example 11

The effect of different amounts of combinations of VYR206 and CBD on various DLBCL and MCL cell lines were evaluated by immunoblot analysis. Immunoblot analysis of cell lysates from various DLBCL and MCL cell lines was performed to show effects on expression of CBi and CB₂ receptors with the expression of β-actin as the control for sample loading (FIGS. 15A-15D), with each cell line identified by respective initials. Northern blotting showed decreasing RNA expression levels of CBi with increased amounts of conjugated CBi inverse agonist VYR206 with predominately no change in CB₂ expression. Moreover, FIGS. 15B and 15C shows a modulation in CB₂ mRNA expression levels with VYR206 and CBD administration.

[00110] Example 12

[00111] Cell viability assays (MTS) were performed to determine the viability of MCL (Mino, Jeko, Sp-53, Maver, Z-138, JMP-1, Rec-1 and PF-1) and DLBCL (CJ, LP, RC, TMD-8, WP, LY- 3, LY-19 and MZ) cancer cell lines in vitro (72 h incubation) in the presence of a cannabinoid receptor ligand, cannabidiol CBD-99, which is a hemp derived cannabidiol from Isodiol International Inc., headquartered in Vancouver, Canada, and has 99% purity. The efficacy of the cannabidiol CBD-99 on the viability of various MCL and DLBCL cell lines is shown in FIGS. 16A and 16B, respectively. The cannabidiol compound CBD-99 used in Example 12 is a non- conjugated (N4 chelator-free) agent.

[00112] Example 13

[00113] Cell viability assays (MTS) were performed to determine the viability of DLBCL and MCL cancer cell lines in vitro (72 h incubation) in the presence of CB i- selective inverse agonists— rimonabant (identified as Rimo) and Bruton's tyrosine kinase inhibitor- Ibrutinib (identified as IBN). The IC50 values of rimonabant and Ibrutinib on the viability of various DLBCL cell lines are shown in FIG. 17, and of various MCL cell lines (Jeko, Mino, PF-4 and PF-5) are shown in FIGS. 18A-18D. All compounds depicted in FIGS. 17 and 18A-18D correspond to non- conjugated (N4 chelator-free) agents. FIGS. 18A - 18D are graphical representations of the effect of the rimonabant and ibrutinib on the viability of MCL cell lines (Jeko, Jeko-R, Mino, Mino-R, PF-4 and PF-5) cell lines' viability. FIG. 18A is a graphical representation of the effect of 25 μΜ of rimonabant and ibrutinib on Jeko and Jeko-R cell lines. FIG. 18B is a graphical representation of the effect of 25 μΜ of rimonabant and ibrutinib on Mino and Mino-R. FIG. 18C is a graphical representation of efficacy of ibrutinib on PF-4 and PF-5 cell lines. FIG. 18D is a graphical representation of the effect of 25 μΜ of rimonabant and ibrutinib on PF-4 and PF-5. Table 3

[00114] Example 14

[00115] Cell viability assays (MTS) were also performed to determine the viability of DLBCL and MCL cancer cell lines in vitro (72 h incubation) in the presence of conjugated CBi-selective inverse agonists— VYR206. IC50 values on DLBCL cell line's viability is shown in Table 3.

[00116] Table 3

Cell line VYR206 ICso (μΜ)

LR 8

LY10 7

LY3 6

U2932 88

LP 13

HBL-1 101

[00117] Example 15

[00118] The efficacy of rimonabant and its cyclam conjugated form, VYR206 on the viability of DLCBL cell lines (LR, MS, RC and DOHH2) and MCL cell lines (Jeko, Rec-1, Mino and JMP- 1) was evaluated with cyclam as the control. FIGS. 19A - 19H are graphical representations of the efficacy of rimonabant and its cyclam conjugated form, VYR206 on the viability of DLCBL cell lines (LR, MS, RC and DOHH2) and MCL cell lines (Jeko, Rec-1, Mino and JMP-1). FIG. 19A is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of LR cell line with cyclam as a control. FIG. 19B is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of MS cell line with cyclam as a control. FIG. 19C is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of RC cell line with cyclam as a control. FIG. 19D is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of DOHH2 cell line with cyclam as a control. FIG. 19E is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of Jeko cell line with cyclam as a control. FIG. 19F is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of Rec-1 cell line with cyclam as a control. FIG. 19G is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of Mino cell line with cyclam as a control. FIG. 19H is a graphical representation of the efficacy of rimonabant and VYR206 on the viability of JMP-1 cell line with cyclam as a control. VYR206 follows the same dose response pattern as the unconjugated rimonabant but with decreased potency. This decreased potency can help decrease the toxicity and other side effects associated with rimonabant. Most of the cell lines presented have some sensitivity to VYR206 where Rec-1 appears to be resistant. This may prove that VYR206 may be selective to certain cell lines.

[00119] Example 16

[00120] Cell viability assays (MTS) were performed to determine the viability of DLBCL (RC) and MCL (Mino) cancer cell lines in vitro (72 hour-incubation) in the presence of CB₂- agonist— CBD alone and in combination with ibrutinib. Results are shown in FIG. 20A. Cell viability assays (MTS) were performed to determine the viability of DLBCL (RC) and MCL (Mino) cancer cell lines in vitro (72 h incubation) in the presence of CBD alone and in combination with zanubrutinib (BGB). Results are shown in FIG. 20B. Cell viability assays (MTS) were performed to determine the viability of DLBCL (RC) and MCL (Mino) cancer cell lines in vitro (72 h incubation) in the presence of CBD alone and in combination with carfilzomib (CFZ). Results are shown in FIG. 20C. Cell viability assays (MTS) were performed to determine the viability of DLBCL (RC) and MCL (Mino) cancer cell lines in vitro (72 h incubation) in the presence CBD alone and in combination with tumorex (TMX). Results are shown in FIG. 20D. Comparative reduction in viability with CBD versus chemotherapeutic agents were demonstrated in FIGS. 20A - 20D where each combination demonstrates a marked reduction in both cell lines, for what appears to be synergistic effect. At the given amounts of CBD and the chosen chemotherapeutic s individually, there was modest reduction in viability, but the combination in each instance caused a synergistic effect on MCL (Mino) and DLBCL (RC) cell lines. This demonstrates combinations of the cannabinoid ligands with chemotherapeutic s can enhance therapeutic effects of the chemotherapeutic compound.

[00121] Example 17

[00122] To determine whether cyclam-conjugated CB₂ agonist CBD and cyclam-conjugated CBi inverse agonist VYR206 at various concentrations resulted in induction of apoptosis, RC cells were treated with increasing concentration of cyclam-conjugated conjugated CBD or VYR206 for 24 hrs. No activation of apoptosis in untreated cells was observed (control not shown). Concentration-dependent increase was detected in cleaved PARP after treatment with cyclam- conjugated conjugated CBD and VYR206. At 12.5 μΜ CBD and 50 μΜ VYR206, there was significant apoptosis induction in cleaved PARP expression. Immunoblot analysis showing the induction of apoptosis through the effects on expression of cPARP are shown in FIG. 21A with expression of β-actin as the control for sample loading in FIG. 21B for cells treated with cyclam- conjugated CB₂ agonist CBD and cyclam-conjugated CBi inverse agonist VYR206 at 6.25, 12.5, 25, and 50 μΜ. Western blotting showed increasing protein expression levels of cPARP with increased amounts of cyclam-conjugated conjugated CBD and VYR206.

[00123] To determine whether different CBD concentrations resulted in induction of apoptosis, BJAB cells were treated with increasing concentration of CBD for 24 hrs. No activation of apoptosis in untreated cells (control not shown). It was observed a concentration-dependent increase in cleaved caspases 3 and cleaved PARP after CBD treatment. At 12.5 μΜ CBD, there was significant apoptosis induction in cleaved PARP expression. The effect of CBD on both cleaved caspase 3 (FIG. 21C) and cleaved PARP expression (FIG. 21D) was analyzed using Western analysis. Expression of the β-actin (FIG. 21E) was used as a control for equal loading.

[00124] Example 18

[00125] Cell viability assays (MTS) were performed to determine the viability of DLBCL (LY19 and LR) cancer cell lines in vitro (72 h incubation) in the presence of CB₂- agonist— CBD in comparison and in combination with conjugated CBi inverse agonist- VYR206. Results are shown in FIGS. 22A and 22B. CBD has mixed affinity for both CBi and CB₂ receptors and had demonstrated generally the same response as VYR206 alone and in combination.

[00126] Example 19

[00127] Genetic profile analysis of DLBCL (RC) cell line for probing for potential molecular marker to develop a target based molecular signature. The expression of genes in untreated and rimonabant treated cells showing the range of under expression (green) to over expression (red) shown in FIG. 27. Genes with significant change in expression in untreated versus treated cells and their functional association are further listed in Table 4.

[00128] Table 4

[00129] Proteomic profile analysis of DLBCL (RC) cell line to demonstrate correlation between most significant proteins is shown in Table 6. These proteins regulate various cellular function ranging from cell adhesion to cell growth and proliferation to apoptosis. For example, Fox03a, known to be associated with the trigger of apoptosis, is shown to have a positive linear relationship with N-Cadherin and RPA32 which are associated with cell adhesion and replication respectively. These correlations provide further insight to potentially understanding the mechanism of cell signaling that is occurring in the DLBCL cell line (RC) and where the cannabinoid receptor activation may be involved.

[00130] Table 6

[00131] Further modifications and alternative embodiments of various aspects of the compositions and methods disclosed here will be apparent in view of this description. Accordingly, this description is to be construed as illustrative only and is for the purpose of teaching those skilled in the art the general manner of carrying out the embodiments. It is to be understood that the forms of the embodiments shown and described here are to be taken as examples of embodiments. Elements and materials may be substituted for those illustrated and described here, parts and processes may be reversed or omitted, and certain features of the embodiments may be utilized independently, all as would be apparent after having the benefit of this description of the embodiments. Changes may be made in the elements described here without departing from the scope of the embodiments as described in the following claims. [00132] The foregoing descriptions of methods, compositions, and results obtained using them are provided merely as illustrative examples. Descriptions of the methods are not intended to require or imply that the steps of the various embodiments must be performed in the order presented. The steps in the foregoing embodiments may be performed in any order. Words such as "then" are not intended to limit the order of the steps; these words are simply used to guide the reader through the description of the methods. Many of the operations may be performed in parallel or concurrently. In addition, the order of the operations may be re-arranged. Various modifications to these embodiments will be readily apparent based on the description provided here, and the generic principles defined here may be applied to other embodiments without departing from the scope of the disclosure.

Claims

CLAIMS What is claimed is:

1. A composition comprising a conjugate of a label, a chelator, and a cannabinoid analog.

2. The composition of Claim 1, wherein the label is a radionuclide.

3. The composition as in any one of the preceding claims, wherein the label is one or more of Technetium-99, Gallium-68, Copper-60, Copper-64, Indium-I l l, Holmium-166, Rhenium-186, Rhenium -188, Yttrium-90, Lutetium-177, Radium-223, or Actinium- 225.

4. The composition as in any one of the preceding claims, wherein the label is configured to facilitate contrast-enhanced imaging, when the composition is administered to a mammal during use.

5. The composition as in any one of the preceding claims, wherein the cannabinoid analog is an immune check point cannabinoid receptor ligand.

6. The composition as in any one of the preceding claims, wherein the cannabinoid analog is a synthetic cannabinoid receptor agonist.

7. The composition as in any one of the preceding claims, wherein the cannabinoid analog is a natural cannabinoid receptor agonist.

8. The composition as in any one of the preceding claims, wherein the cannabinoid analog is an aminoalkylindole.

9. The composition as in any one of the preceding claims, wherein the cannabinoid analog is a cannabinoid receptor inverse agonist.

10. The composition as in any one of the preceding claims, wherein the cannabinoid analog belongs to the group consisting of anandamide (AEA), 2-arachidonoylglycerol (2-AG), noladin ether, virodhamine and N-arachidonylodopamine (NAD A).

11. The composition as in any one of the preceding claims, wherein the cannabinoid analog is a diarylopyrazole.

12. The composition as in any one of the preceding claims, wherein the chelator is an aminated or an acid chelator.

13. The composition as in any one of the preceding claims, wherein the chelator is a cyclam.

14. The composition as in any one of the preceding claims, wherein the cannabinoid analog is cyclam- l'-acetyl-[N-(Piperidin-l-yl)-5-(4-chlorophenyl)-l-(2,4-dichlorophenyl)-4- methyl- lH-pyrazole-3-carboxamide] .

15. The composition as in any one of the preceding claims, wherein the cannabinoid analog is cyclam-l'-propyl-[N-(Piperidin- l-yl)-5-(4-chlorophenyl)- l-(2,4-dichlorophenyl)-4- methyl- lH-pyrazole-3-carboxamide] .

16. A method for diagnosing a medical condition in a human subject, the method comprising administering to a human subject an effective amount of the composition as in any one of the preceding claims.

17. A method of imaging a plurality of cancer cells, the method comprising:

administering to a human patient an effective amount of the composition of any one of claims 1-15.

18. A method for treating a cancer amenable to treatment with a cannabinoid, the method comprising:

administering to a human subject in need thereof an effective amount of the composition of any one of claims 1- 15.

19. A method for treating a neurologic disorder amenable to treatment with a cannabinoid, the method comprising:

20. A method for alleviating pain in a human subject, the method comprising:

21. A kit for imaging cells, comprising:

a predetermined quantity of a conjugate of a chelator and a medical cannabinoid analog; and

a predetermined quantity of an imaging agent.

22. The kit of Claim 21, further comprising a tin-containing reducing agent.