Join Date: Oct 2008
Location: Ditchweed City
easy synthesis of HU-331, HU-336, HU-345
The Beam test for CBD results in some oxidized-CBD-things that can be isolated. One of these products has been given a code name by the research facility funding the investigation, and so is forever HU-331.
Both the pure chemical and the impure reaction mixture have been evaluated for their action against human cancer cells. It is not exactly known how to use this to provide a benefit to humans with cancers, but it is safe to say that CBD, CBN, and the THCs are less effective. A problem with using cannabinoids against cancers is providing a high dose to the site, and unfortunately this new drug is like CBD and THC in being approximately insoluble in water. However, unlike many synthetic cannabinoids, HU-331 is at least quite easy for even a non-chemist to make. It's not at all clear if smoking or eating these substances does anything.
There are a few papers on this in the science journals, and the inventors (Mechoulam et al.) received US patent 8,497,299 in 2013 for potential commercial application. No one but them can legally make money off this, so no one else is going to be doing any trials on humans. Your doctor can't get this stuff and wouldn't inject you with it if you had some, even if you had a month to live.
Mechoulam would probably not be happy with people making and using or providing these chemicals outside of a rigorous scientific program, but we are talking about dying people with little to lose. It's not clear to me what the shelf life of these chemicals are but my guess is not forever, and they should probably be stored in a freezer. CBD and/or its metabolites and other cannabinoids affect the potency of other drugs by affecting the pathways that break them down, so some care is necessary when using and especially when recommending the cannabinoids while other drugs are being taken.
Chief Idiot of Florida Pam Bondi, as attorney general, in order to prevent an epidemic of sanity in that state, years ago declared an emergency and made illegal HU-331 and other non-psychoactive synthetic chemicals being explored by scientists at the time, and her biggest fan Wisconsin followed suit. Calling something a cannabinoid or phenethylamine is the kiss of death for it in FL and I hope she made the only cure for her cancer forbidden to even research.
The patent merges what has been written on the subject up to then into a convenient single source. Selected sections follow. Where they confusingly say ethanolic 5% KOHaq, they mean 500 mg. of KOH dissolved in 10 ml. ethanol, with no water other than the 10% normally in KOH. NaOH should work just as well as KOH, but it doesn't dissolve in alcohols as easily. The preparations using CuCl in acetonitrile are almost as easy to do. Other examples in the patent use BTIB - you're less likely to obtain this or its precursors, so they're not included here.
As described in the Examples, the five quinonoid derivatives of cannabinoids (or cannabinoic quinones), which were synthesized by the inventors, were tested for their anti-proliferative activity on human cancer cell lines. These five compounds had anti-proliferative activity in cell lines which originated from various types of cancer, more specifically from lymphomas, mammary gland (breast), prostate, lung, glioblastoma, and colon.
Thus, the pharmaceutical composition of the invention may be for the treatment of hyperproliferative disorders such as carcinomas, lymphomas, melanomas, glioblastomas and sarcomas. Alternatively, the pharmaceutical composition of the invention may be for the treatment of a non-malignant hyperproliferative disorder, for example psoriasis.
The pharmaceutical composition of the invention may optionally further comprise carriers, additives and diluents.
The preparation of pharmaceutical compositions is well known in the art and has been described in many articles and textbooks, see e.g., Remington's Pharmaceutical Sciences, Gennaro A. R. ed., Mack Publishing Co., Easton, Pa., 1990, and especially pp. 1521-1712 therein.
Compounds with anti-cancer activity are often difficult to solubilize. HU-331 exerted a striking anti-tumor effect in vivo, following either sub-cutaneous (s.c.) or intra-peritoneal (i.p.) administration, when solubilized in ethanol:Emulphor®:PBS (1:1:18), suggesting that this solvent enabled its bioavailability at the cancer site.
Thus, in preferred embodiments, the pharmaceutical compositions of the invention comprise a pharmaceutically acceptable vehicle or carrier, particularly a mixture of ethanol:Emulphor®:phosphate buffered saline (PBS), at 1:1:18 v/v ratio. Emulphor® is a polyoxyethylated vegetable oil, a high-molecular-weight ether sulphate which is an anionic emulsifier. A preferred buffer is PBS, but any physiologically compatible buffer may be used, e.g. Hepes, Tris-buffered saline. Poly(ethylene glycol) and cyclodextrins of various types, like alkylated beta-cyclodextrin, for example, are also suitable carriers.
The pharmaceutical compositions of the invention, comprising as active agent cannabinoid quinones, may also be used in the treatment of conditions such as inflammations, autoimmune diseases (for example multiple sclerosis) and infections, particularly infections caused by infectious agents like bacteria (Gram positive and Gram negative, mycobacteria, etc.), protozoa and fungus.
The compositions of the invention can be administered in a variety of ways. By way of non-limiting example, the composition may be delivered intravenously, or into a body cavity adjacent to the location of a solid tumor, such as the intraperitoneal cavity, or injected directly into or adjacent to a solid tumor. Intravenous administration, for example, is advantageous in the treatment of leukemias, lymphomas, and comparable malignancies of the lymphatic system.
As a preferred route the composition of the present invention may be administered via subcutaneous or intradermal injections in proximity to the tumor, via intratumor or intraperitoneal injection.
The composition of the invention may also be delivered in the form of gelatin capsules, wherein the active agent will be dissolved in poly(ethylene glycols) of the lower molecular weights, suitable for the preparation of said capsules.
The quinones described herein proved to be powerful antitumor agents in vivo. As shown in the Examples, nude mice which developed a tumor after the injection of tumor cell lines, had the tumor size significantly decreased after administration of HU-331.
Hence, in another aspect, the present invention relates to a method for the treatment of a hyperproliferative disorder, malignant or non-malignant, comprising administering a therapeutic effective amount of a cannabinoic quinone or of a pharmaceutical composition thereof to a subject in need. Preferred cannabinoic quinones are those represented by one of formulae (I), (II), (III), (IV) and (V) as described above. As detailed below, most preferred cannabinoic quinones are HU-331, HU-336, HU-345, HU-395 and HU-396, and particularly HU-331.
Said therapeutic effective amount, or dosage, is dependent on severity and responsiveness of the disease state to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. The therapeutic effective dosage may be determined by various methods, including generating an empirical dose-response curve, predicting potency and efficacy of a congener by using quantitative structure activity relationships (QSAR) methods or molecular modeling, and other methods used in the pharmaceutical sciences. Optimal dosing schedules may also be calculated from measurements of drug accumulation in the body of the patient. Persons of ordinary skill can easily determine optimum dosages, dosing methodologies and repetition rates. In general, dosage is from 0.01 ug to 100 g per kg of body weight, and may be given once or more daily, weekly, or monthly. Persons of ordinary skill in the art can easily estimate repetition rates for dosing based on the patient's response to the active agent.
The cannabinoic quinone to be used in the method of treatment of the invention is preferably one of the componnds synthesized by the inventors, i.e., HU-331, HU-336, HU-345, HU-395 and HU-396.
Malignant hyperproliferative or proliferative disorders that may be treated by the method of the invention are, for example, carcinoma, lymphoma, melanoma, glioblastoma or sarcoma.
As used herein, "tumor", "cancer" and "malignant hyperproliferative (or proliferative) disorder" all relate equivalently to a hyperplasia of a tissue or organ. If the tissue is part of the lymphatic or immune systems, malignant cells may include non-solid tumors of circulating cells. Malignancies of other tissues or organs may produce solid tumors. In general, the compounds of the present invention as well as the method of the present invention may be used in the treatment of non-solid and solid tumors.
Thus, the present invention provides a cannabinoic quinone, preferably HU-331, or compositions comprising the same, to be used in the treatment of hyperproliferative disorders, particularly colon cancer, lymphoma and breast cancer.
HU-331 displayed a marked anti-cancer activity not only in vitro but also in vivo, in experiments where nude mice received a subcutaneous inoculation of HT-29 colon carcinoma cells (see Example 3). The administration of HU-331 at a concentration that did not have observable adverse effects on the hosts resulted in significant inhibition of the growth of the tumor cells when injected either intraperitoneally (i.p.) or subcutaneously (s.c.) into the region of the tumor graft.
Thus, a major advantage of the cannabinoic quinones described herein (HU-331, HU-336, HU-345, HU-395 and HU-396) is that at concentrations in which they display a medicinal activity, in particular an anti-proliferative activity, these compounds do not have any measurable side-effects, such as weight loss, or any toxic cardiac effect. Therefore, HU-331, HU-336, HU-345, HU-395 and HU-396 may be used as anti-tumor or anti-cancer agents.
Hence, the present invention provides an alternative for cancer treatment, through the use of the cannabinoic quinones described in the invention, with clear benefits for the patient in need of said treatment.
In a further aspect, the present invention provides a method for the treatment of conditions selected from the group consisting of inflammations, autoimmune diseases (in particular multiple sclerosis), and infections, particularly infections caused by infectious agents such as bacteria (including gram positive and gram negative bacteria, mycobacteria, etc.), protozoa and fungus, wherein said method comprises administering a therapeutic effective amount of a cannabinoic quinone or of a pharmaceutical composition thereof to a subject in need. Preferred cannabinoic quinones are those represented by one of formulae (I), (II), (III), (IV) and (V) as described above. Most preferred cannabinoic quinones are HU-331, HU-336, HU-345, HU-395 and HU-396.
Most importantly, the cannabinoids herein described do not demonstrate any measurable psychotropic effects. The cannabinoic quinones were not able to bind to the cannabinoid receptors CB1 and CB2, up to 15 uM. For comparison, the binding constant of tetrahydrocannabiol is 50 nM.
In one embodiment, HU-331 is the cannabinoic quinone to be used in the method of treatment of the invention, particularly when the proliferative disorder to be treated is colon cancer. Alternatively, this compound is also preferred for the treatment of lymphoma and/or breast cancer.
As shown in Example 3, HU-336, HU-345 and HU-396 required higher concentrations to display their anti-proliferative activity.
HU-336 (4), at a concentration of 12.5 ug/ml or higher, inhibited 50% or more the growth of all the cells tested. The growth of SNB-19 cells was inhibited by HU-336 only at a concentration of 100 ug/ml.
HU-345 (6) was a more potent anti-cancer reagent than HU-336. Raji lymphoma cells growth was inhibited by over 50% at a concentration of 6.25 ug/ml, while that Jurkat lymphoma cells and DU-145 prostate cancer cells were inhibited by a concentration of 12.5 ug/ml. At concentrations of 25.0 ug/ml of HU-345, all cell lines tested were inhibited.
Interestingly, the growth of cell lines SNB-19 and DU-145 was more effectively inhibited by HU-336 and HU-345 than by HU-331.
Thus, in another embodiment, HU-336 and/or HU-345 are to be used in the method of the invention, particularly when the hyperproliferative disorder to be treated is prostate cancer or glioblastoma.
The mechanism of the anti-cancer activity of cannabinoic quinones is still unclear. However, a number of mechanisms have been suggested by which quinones may exert cell damage [Ollinger., K. and Kagedal., K. (2002) Subcell Biochem. 36, 151-70]. These include redox cycling, DNA damage and inhibition of topoisomerase, protein damage and lipid peroxidation. Similar mechanisms were shown to mediate the antitumor effects of adriamycin and daunorubicin, which have been in clinical use for the treatment of solid tumors for over 30 years [Gewirtz, D. A. (1999) Biochem Pharmacol. 57, 727-41]. The present study clearly indicates that cannabinoic quinones possess a high potential for development into anticancer drugs that may prove effective not only against lymphoma cells but also against solid tumors, with no undesired effects.
Oxidation of cannabidiol (CBD) to HU-331 with KOHaq
CBD (1 g, 3.18 mmole) was dissolved in 90 ml petroleum ether (40-60° bp) and 5% KOHaq in ethanol (10 ml, 8.77 mmole) was added. The reaction mixture was stirred at 0° C. in an open beaker for 3 hours, and after, 25 ml of 5% HCl was poured into it. The organic layer was washed with sodium bicarbonate and water and dried (MgSO4). Removal of the solvent under reduced pressure yielded a glassy oil (1.08 g). HU-331 was eluted on [silica gel]column chromatography with petroleum ether-ether (95:5). After crystallization from heptane, 211 mg (0.64 mmole, 20.2% yield) of large brown crystals were obtained. m.p. (melting point): 50-51° C.
Oxidation of delta8-THC to HU-336 with CuCl
To a solution of d8-THC (104 mg, 0.33 mmole) in 0.9 ml acetonitrile (ACN) CuCl (5.5 mg, 0.056 mmole) was added. A thin current of air was bubbled through the mixture for 1.5 h after which 50 ml ether was added. The reaction mixture was washed with H2O, dried (MgSO4) and concentrated. The yellowish oil obtained was purified by column chromatography using pet. ether-ether (95:5) solution. HU-336 (33 mg, 0.1 mmole, 30.5% yield) was obtained as a yellow oil and crystallized from heptane to obtain very thin yellow needles.
Oxidation of CBN to HU-345 with CuCl
To a solution of CBN (95 mg, 0.31 mmole) in 0.9 ml ACN CuCl (10.8 mg, 0.11 mmole) was added. A thin current of air was bubbled through the mixture for 6 h after which 50 ml ether was added. The reaction mixture was washed with H2O, dried (MgSO4) and concentrated. The red oil obtained was purified by column chromatography using a petroleum ether-ether (93:7) solution. HU-345 (15 mg, 0.046 mmole, 15% yield) was obtained as red oil and crystallized from heptane to obtain large red crystals. m.p.: 81-82° C.
In the clinical field, the practical application of these substances must be awaited with the usual necessary patience.
- Roger Adams
February 19, 1942