WO2001058445A1 - Therapy with cannabinoids in the treatment of cerebral tumor - Google Patents

Therapy with cannabinoids in the treatment of cerebral tumor Download PDF

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WO2001058445A1
WO2001058445A1 PCT/ES2000/000450 ES0000450W WO0158445A1 WO 2001058445 A1 WO2001058445 A1 WO 2001058445A1 ES 0000450 W ES0000450 W ES 0000450W WO 0158445 A1 WO0158445 A1 WO 0158445A1
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cannabinoids
cannabinoid
malignant
tumors
natural
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PCT/ES2000/000450
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Spanish (es)
French (fr)
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Manuel Guzman Pastor
Cristina Sanchez Garcia
Ismael Galve Roperh
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Universidad Complutense De Madrid
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/335Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. cannabinols, methantheline
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/045Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05Phenols
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
    • A61K31/435Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47Quinolines; Isoquinolines
    • A61K31/473Quinolines; Isoquinolines ortho- or peri-condensed with carbocyclic ring systems, e.g. acridines, phenanthridines
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
    • A61K31/535Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/53751,4-Oxazines, e.g. morpholine
    • A61K31/53831,4-Oxazines, e.g. morpholine ortho- or peri-condensed with heterocyclic ring systems

Abstract

The therapy with cannabinoids in the treatment of cerebral tumors involves (intracranial or systematic) administration of (natural or synthetic) cannabinoids to (human or non-human) mammals having cerebral tumors. Activation of the specific receptors of the cannabinoids leads to selective death of the transformed cells. Regression or eradication of the cerebral tumors is achieved without any significant side-effects.

Description

Title

THERAPY cannabinoids ELTRATAMIENTO OF BRAIN TUMORS

TECHNICAL FIELD OF THE INVENTION

The present invention is in the field of brain tumor therapy

OBJECT OF THE INVENTION

The present invention as stated herein refers to the therapeutic use of cannabinoids for the treatment of brain tumors therapies used today to treat these tumors (surgery, radiotherapy, chemotherapy, immunotherapy, gene therapy) are ineffective or at best palliative the invention implies a technically simple approach lacking noticeable side effects and highly effective for the treatment of the malignant tumors including brain (glioblastomas)

Background

In brain tumors affecting humans glioblastomas are the most common (1 per 50,000 persons per year) malignant (nearly 100% mortality) and fastest evolving (life expectancy of weeks / months after diagnosis ) Today, the treatment of glioblastomas is generally ineffective or merely palliative, and involves techniques such as surgery, radiotherapy, chemotherapy and immunotherapy (Louis, DN & Gusella, JF, Trenas Genet. 11, 412-415 1995 Avgeropoulos, NG Batchelor, TT, Oncologist 4, 209-224, 1999) in addition, gene therapy has begun to be used as an experimental treatment for glioblastomas, although so far has provided few positive results (Martuza, RL, Nature Med 3, 1323, 1997) the already unlikely success of these therapeutic approaches often also be complicated by factors such as rapid growth, remarkable heterogeneity, the high level of infiltration and extreme resis tence chemotherapy exhibiting glioblastomas (Maintz, D et al, J Neuropathol Exp Neurol 56, 1098-1 104, 1997, Mason, W, Louis, DN & Cairncross, JG, J Clin Oncol 15, 3423-3426, 1997, Martuza, op cit, Avgeropoulos & Batchelor, op cit) seems clear therefore that is highly recommended to develop alternative therapies for the treatment of brain tumors cannabinoids are compounds owe their name to that they are synthesized by the plant Cannabis sativa L These compounds, including the Δ 9 -tetrahιdrocannabιnol (THC) stands out for its high potency and abundance, are responsible for the central and peripheral effects of marijuana (Pertwee, RG, Pharmacol Ther 74, 129 to 180.1997, Felder , CC & Glass, Pharmacol Toxicol M Annu Rev 38, 179-200, 1998) cannabinoids C sativa (Fig 1) carry out their effects because they are similar to certain molecules produced by animals (including humans) probably dese mpeñan important functions in the nervous system Such molecules are called therefore cannabmoides endogenous or endocannabinoids, and anandamide (= araquιdonoιletanolamιda) is the main representative (Di Marzo, V, Melck, D, Bisogno, T & De Petrocellis, L, Trends Neurosci 21, 521-528 1998, Martin, BR, Mechoulam, R & Razdan, RK, Ufe S 65, 573-595, 1999) Moreover, have been obtained in the laboratory compounds mimic the action of natural cannabinoids but with a much higher power are called synthetic cannabinoids one of whose representatives WIN-55212-2 is (Fig 2) (Pertwee, op cit Barth, F, Expert Opm Ther Patents 8, 301-313, 1998) cannabinoids both natural and synthetic, act by binding to specific membrane receptors (cannabinoid receptors or CB type), of which two different subtypes CB Í and CB 2 (Pertwee, op cit, Howlett, is nowadays known a er al, in the lUPHAR C ompendium of Receptor Characteπzation and Classification, eds Godframd, T, Humphrey, P Ruffolo, R & Vanhoutte, P, lUPHAR Media, pp 97-104, 1998) Not all tissues in the body possess these receptors are mainly located in the nervous system, and why mayoπtaπamente cannabinoids exert their effects on the brain (Pertwee, op at, Childers, SR & Breivogel, CS, Drug Alcohol Depen 51, 173-187, 1998) There is now a large number of studies on possible therapeutic applications of cannabinoids Moreover, today and doctors are allowed in the Remo Kingdom and in various states of the United States prescribe THC or certain synthetic cannabinoids as appetite stimulants and inhibitors of vomiting in patients with AIDS or cancer treated chronically with chemotherapy (Gπnspoon, L & Bakalar, JB, JAMA 273, 1875 to 1876.1995, Voth & Schwartz E, R, Ann Intern Med 126, 791-798, 1997) potential therapeutic uses d and cannabinoids include the following (a) as analgesic agents has been shown to act highly effective in mitigating acute and chronic pain, (b) as agents that reduce motor activity being tested today in the treatment of disorders associated with Parkinson's disease, Huntington's chorea and multiple sclerosis, (c) as anticonvulsants application in the treatment of epilepsy is being studied, (d) as agents that decrease intraocular pressure may be used in the treatment glaucoma (Voth & Schwartz, op cit, Manzanares, J er a /., Trends Pharmacol Sci 20, 287-294, 1999, Pop, E, Curr Opm CPNS Invest Drugs 1, 587-596, 1999, Sanudo-Pena, MC, Tsou, K & Walker, JM, Ufe Sci 65, 703-713, 1999) Some of these potential therapeutic uses of cannabinoids have already been patented (see for example US4189491, US5939429, WO9711668, WO9832441 and WO9957106)

One of the most intriguing and unexplored cannabinoid effects is its ability to inhibit the growth of transformed cells in vitro has thus been shown that several cannabinoids inhibit the proliferation of tumor cells MCF-7 (De Petrocellis, L et al , Proc Natl Acad. Sci. USA 95, 8375-8380, 1998), glioblastoma cells C6 (Sánchez, C., Galve-Roperh, I, Canova, C., Brachet, P & Guzman, M, FEBS Lett 436, 6 -10, 1998) and prostate tumor cells PC-3 (Ruiz, L, Miguel, A & Diaz-Laviada, I, FEBS Lett 458, 400-404, 1999) However, these findings in cell culture systems they have never been so far observed in vivo, so its biomedical significance is unknown

DESCRIPTION OF THE INVENTION

The present invention makes use first of cannabinoids for the treatment of brain tumors, and is based on our original observations that cannabinoids induce a marked regression (leading to a longer life) and even eradication (implying the cure) of ghoblastomas in laboratory animals This invention involves the use of a technically simple therapy lacking noticeable side effects and, more importantly, very effective for the treatment of brain tumors, which as previously mentioned may not be treated today successfully with other techniques or compounds experiments which have led to the present invention are detailed below

Antitumoral action of cannabinoids in rats

Injection of C6 glioblastoma cells in the rat brain is widely used as an experimental model of malignant brain tumor (Barth, RF, J Neur∞ncol 36, 91-102, 1998) C6 glioblastoma cells were inoculated directly into the brain Wistar rats, and the tumors visualized by magnetic resonance All animals left untreated died was uniformly 12-18 days after inoculation of the cells (Fig 3a). To evaluate the antitumoral potential of the cannabinoids, 12 days after inoculation of cells was administered to a group of animals for 7 days THC or WIN-55,212-2 through a cannula located at the site of inoculation treated animals cannabinoids had a significantly longer life than control animals (Fig 3a). Thus, administration of cannabinoids managed to increase survival to 19-35 days in 9/15 of animals (treatment with THC) or 19-43 days in 4/15 of animals (treatment with WIN-55212-2) Moreover, cannabinoids eradicated completely the tumor in 3/15 of animals (treatment with THC) or 5/15 of animals (treatment with WIN 55,212-2-) Figure 3b an image of magnetic resonance of one of the animals cured with THC it is shown, after administration of the cannabinoid the tumoral mass disappeared completely and instead a residual hipomteπsa area interpreted as a fibrous scar at the site of inoculation No recurrence was observed in 8 animals cured observed caπnabmoides

antitumor action of cannabinoids in immunodeficient mice

To discern whether the antiproliferative action of cannabinoids was due to a direct effect on tumor cells or an indirect effect mediated by an immune response, C6 glioblastoma cells were inoculated subcutaneously in mice deficient in RAG recombinase-2 (RAG - 2 ' 1 '), which lack mature T and B lymphocytes (Shinkai, Y et al, Ce // 68, 855- 867 , 1992) As shown in Fig 4a, the size of the tumors was extraordinarily smaller in animals treated with THC or WIN-55,212-2 than in control animals in Fig 4b examples of tumor bearing mice and tumors are shown after treatment with or without cannabinoids dissected for 7 days

Sureness treatment in vivo cannabinoid were then examined the possible side effects of treatment with cannabinoids Rats without tumor and treated with cannabinoids did not see completely unaffected survival (Fig 3a) As in the 8 animals mentioned above whose tumors were eradicated with cannabinoids, a detailed analysis by magnetic resonance of all animals without tumor revealed that treatment with cannabinoids did not produce any signs of damage by necrosis, edema, infection, inflammation or trauma to rule out the possibility that cannabinoids were toxic to nerve cells in division, stainings TUNNEL were performed in subventπcular area rat brains, which continues to proliferate in the adult animal the administration of cannabinoids not only not produce any significant apoptotic effect in the in vivo brain, but also the slight marking observed in the caud ado putamen of control animals was not apparent in animals treated with cannabinoids

Both animals without tumor and tumor-bearing, cannabinoids did not induce any significant alteration of behavioral parameters such as motor coordination and physical activity intake of water and food, as well as weight gain, nor were affected by cannabinoids Likewise, in the blood biochemical parameters (glucose, urea, uric acid, creatinine, cholesterol, bilirubin) and markers of tissue injury (alanine and aspartate ammotransferasas, γ-glutamyltransferase, creatine kinase, lactate dehydrogenase) were unaffected or over the period of 7 days of administration or up to 2 months after completion of treatment with cannabinoids data of other authors support the idea that cannabinoids are not only non-toxic compounds for nerve cells, but even protect them toxic stimuli such as glutamaérgicos agonists (Skaper, SD et al, Pro c Sα Natl Acad USA 93, 3984-3989, 1996, Shen, M & Thayer, SA, Mol Pharmacol 54, 459-462, 1998), oxidizing agents (Hampson, AJ, Gπmaldi, M Axelrod, J & Wink, D , Proc Natl Acad Sci USA 95, 8268-8273, 1998) and ischemia (Nagayama, T er a /, J Neurosci 19, 2987-2995, 1999)

Pharmacological characterization of the antitumoral action of cannabinoids

They were carried out experiments to characterize pharmacologically the cannabinoid of C6 glioblastoma cells induced death in culture Agonists synthetic high power as

WIN 55,212-2-, CP-55,940 and HU-210 induced the death of these cells in lower doses than THC, as expected from their greater affinity for cannabinoid receptors (Pertwee, op at) Thus, after 5 days exposure to cannabinoids the viability of C6 glioblastoma was reduced by 50% at concentrations of 20 nM WIN-55, 212-2, 45 nM CP- 55,940, 10 nM HU-210 and 480 nM THC (n = 4) Ni SR141716 (a selective antagonist of CB nor SR144528 (a selective antagonist of CB2) (Shire, D et al, Life Sci 65, 627-635, 1999) were able separately to prevent the induced cell death THC No But when the two antagonists were added together to the incubations an effective prevention by THC (Fig 5a) Accordingly induced cell death was observed, the Western blot analysis showed that C6 glioblastoma cells expressed both the CBi receptor as CB 2 (Fig 5b)

Application of the invention in other cases The experiments which led to the present invention have been carried out with rats and mice as tumor-bearing animals, however, both the experimental design used as the similarity exhibiting brain tumors in different mammals (RF Barth, op cit), the invention is applicable to the treatment of brain tumors in other mammals, including humans

The experiments leading to the present invention have been carried out with glioblastomas as a model of brain tumor However, the experimental design was used for induction and treatment of tumors, the invention is applicable to treatment of other tumors brain, for example medulloepitheliomas, medulloblastomas, neuroblastomas, germinoma, embryonal carcinomas, astrocytomas, astroblastomas, ependymomas, omas oligodendrog, plexales, neuroepitehomas carcinomas, pineoblastomas, ependymoblastomas, neuroectodermal tumors, malignant meningiomas, chondrosarcomas, meningeal sarcomatosomas, malignant melanomas or schwannomas malignant experiments leading to the present invention have been carried out with two paradigmatic cannabinoids, a natural one (THC) and a synthetic (WIN 55,212-2-) in a preferred embodiment of the invention the cannabinoid is used with more potent antiproliferative effect a certain Tumo r However, as the antiproliferative effect of these compounds is mediated by cannabinoid receptors (receptors CB type, Howlett et al, op αt), the invention is applicable to any other agonist of these receptors, both cannabmoides C sativa (eg Δ 8 -tetrahιdrocannabιnol, cannabinol, cannabidiol) (Fig 1) as synthetic cannabinoids (such as HU-210, CP-55,940, CP-50.556) (Fig 2) (Pertwee, op αt, F Barth, op cit) also included in this section medicinal products containing in its composition any cannabinoid

The experiments leading to the present invention have conducted intratumorally administering the cannabinoid In a preferred embodiment of the invention this will be the chosen route of administration, since it allows a high accessibility of the cannabinoid to the tumor However, since the action cannabinoid is direct on the tumor and does not appear to substantially affect peripheral systems, the route of administration may also be systemic, such intrapeπtoneal, intravenous or oral

The experiments leading to the present invention have been carried out by continuous administration of a dose of cannabinoid for a set time In a preferred embodiment of the invention these parameters will be optimized depending on the specific requirements of the treatment status of the patient, size and location of the tumor, number of tumors, etc. Thus, for example, the application mode may be continuous (preferred mode) or sequential in one or several doses per day This will obviously vary the doses administered compound and the total time treatment

BRIEF DESCRIPTION OF THE FIGURES

Figure 1

Chemical formula of the main cannabmoides C sativa

Figure 2

Chemical formula of the main synthetic cannabinoids

Figure 3

Antitumoral action of cannabinoids in rats (a) Survival curves for rats with brain tumors were induced in 45 rats glioblastomas (day 0), 15 animals were not treated with cannabinoids (---), while another 15 were treated with THC (-) and 15 WIN-55212-2 with () between 12 and 19 animals treated with cannabinoids lived significantly longer than controls (P <0 01 by the log-rank test) is thus given the same THC and WIN 55,212-2-5 rats each in which no tumor was induced (- -)

(B) Magnetic resonance image in axial projection (top) and coronal (bottom) brain of a rat before (left) and after

(right) treatment with THC A 100 mm3 glioblastoma (arrow) was eradicated by 500 mg of THC The image was taken 7 days after completion of treatment with THC

Figure 4

Antitumoral action of cannabinoids in immunodeficient mice (a) Glioblastomas were induced in 18 mice When tumors reached the desired size (day 0) 6 animals were treated with vehicle (o), while another 6 were treated with THC (•) and another 6 with WIN-55,212-2 (z) for 7 days the size of tumors in animals treated with cannabinoids was significantly smaller than controls at all times (P <0 01 by the test Student f) ( b) Examples of glioblastomas in mice (top) and dissected

(Bottom bar 1 cm) after treatment for 7 days with vehicle, THC or WIN-55,212-2 (WIN)

Figure 5

Involvement of cannabinoid receptors in cell death

(a) C6 glioblastoma cells were cultured for 5 days in the absence or presence of 1 .mu.M THC, 1 uM SR141716 (SR1) and / or 1 .mu.M SR144528 (SR2) (n = 6) * Sιgnιfιcatιvamente different from incubations without additions ( P <0 01 r by Student test)

(b) Presence of cannabinoid receptors CB T and CB 2 in C6 glioblastoma cells Detection of receptors by Western or / or performed with antibodies specific for each of the two receivers

Embodiment of the invention

The present invention is further illustrated by the examples set forth below

E | emplo 1

Glioblastomas healing in rats

Male Wistar rats were anesthetized (250-300 g body weight)

3% isoflurane in a mixture of oxygen (0 8 l / mm) and protoxide (0 4 l / mm)

Were prepared 5x10 6 C6 glioblastoma cells in 100 ul buffered saline (PBS) supplemented with 0 1% glucose and stereotactically injected into the fronto-paπetal lobe of the right hemisphere (4 mm to the right of bregma, 4 5 mm depth from the cranium) (Izquierdo, M et al, Gene Ther 2, 66-69, 1995) the rats received dexamethasone (2 mg / l) and tetracycline (75 mg / kg body weight) in water for 3 days before and 7 days after inoculating the cells a thorough monitoring of the tumors was performed by magnetic resonance methods described by other authors (Izquierdo et al, op αt, Cortes ML, de Felipe P, Martin , V, Hughes MA & Izquierdo, M, Gene Ther 5, 1499-1507, 1998)

The administration of cannabinoids to rats began 12 days the after cell inoculation At this time, the average tumor size was 70 mm3 (interval 25-100 mm3) as estimated by magnetic resonance (Izquierdo et al, op cit, Cortés, de Felipe, Martin Hughes & Izquierdo, op αt) cannabinoids were administered by a cannula placed at the site of tumor inoculation and fixed to the skull with dental cement, a small thyme stainless steel ensured cannula and cement The cannula was connected dental subcutaneously via a catheter to an osmotic mini-pump (Alzet 2001) which operated at a flow rate of 1 ul / h for 7 days The osmotic pump was filled with 500-2500 mg of THC or 50-250 ug WIN 55,212-2-in 200 ul of PBS supplemented with 5 mg / ml bovine serum albumin (BSA) and dialyzed des pidizada cannabinoid doses used depended on the characteristics of tumor to be treated, higher doses are emple aron for large, dense and invasive tumors

As shown in Fig 3a, all animals left untreated died uniformly 12-18 days after cell inoculation the animals treated with cannabinoids had a significantly longer life than control animals Moreover, cannabinoids eradicated completely the tumor in a significant percentage of animals in Fig 3B, a magnetic resonance image of one of the cured THC following administration of the cannabinoid animal tumor mass disappears completely shown, and a residual hypointense area is interpreted observed as a fibrous scar at the site of inoculation No recurrence was observed in animals cured with cannabinoids

example 2

Qlioblastomas healing in mice mmunodeficientes

Tumors were induced in RAG-2 'mice "by subcutaneous inoculation of 5x10 6 cells ghoblastoma C6 in 100 .mu.l of PBS supplemented with 0 1% glucose About 10 days later when average tumor volume was 250 mm 3 (range 200-300 mm 3), the animals were divided randomly into 3 groups and injected with vehicle for 7 days,

500 mg of THC or 50 μ g of WIN 55,212-2-day in 100 .mu.l of PBS supplemented with 5 mg / ml of delipidized and dialyzed BSA Dimensions of the tumors were measured with a caliper and its volume was calculated as (4π / 3) x (width / 2) 2 x (longιtud / 2)

As shown in Fig 4a, the size of the tumors was extraordinarily smaller in animals treated with THC or WIN

55,212-2 than in control animals in Fig 4b examples of tumor bearing mice and tumors dissected after treatment are shown with or without cannabinoids for 7 days

Eiemplo 3

Implication of cannabinoid receptors in cell death qlioblastoma were cultured at 37 ° C and 5% CO2 C6 glioblastoma cells in medium

F12 supplemented with fetal calf serum 10%, 24 h before the start of the experiment the cells were transferred to medium F12 serum-free and supplemented with insulin (5 ug / ml), transfernna (5ug / ml), sodium selenite (5 .mu.g / ml) and delipidized and dialyzed BSA (10 mg / ml) medium every 48 h renewed cell viability was determined by the MTT method (Sanchez, C, Galve-Ropem, I, Canova, C., Brachet, P & Guzman M, op ​​at) As shown in Fig 5a, THC was capable of inducing the death of C6 glioblastoma cells also when added simultaneously to the medium SR141716 (a selective antagonist of CBi) and SR144528 (a selective antagonist of CB 2) induced cell death is prevented by THC

To verify that both receptors were present in the C6 cells the cells were washed with PBS, the plates scraped in lysis medium and the particulate fraction was obtained by centrifugation at 40 OOOg for 60 min (Sanchez, C, Galve-Roperh, I, Canova, C., Brachet, P & Guzman, M, op ​​at) the samples to gel electrophoresis poliacπlamida were subjected to sodium dodecyl sulfate and the proteins were transferred from gels to nitrocellulose membranes membranes were blocked with BSA deshpidizada and dialyzed 1%, and incubated with an antibody to residues 1-14 of the rat CBi receptor (diluted 1 5000) or with an antibody to residues 350-361 of the CB2 human receptor (diluted 1 2000) the samples were finally subjected to developing with a "kit" electrochemiluminescence (Amersham, Bucks, UK) as shown in Fig 5b, C6 glioblastoma cells expressed both receptor CB Í as the CB 2

Eiemplo 4

Sureness treatment with cannabinoids m cannabmoides vivo (2500 g of THC or 250 mg of WIN-55,212-2) to tumor free rats were administered for 7 days as described above were then sacrificed rats and their brains fixed with 4% paraformaldehyde in PBS death by apoptosis it was determined in brain sections of 40 microns thickness using a "kit" TUNEL staining according to the supplier's instructions (Boehπnger, Mannheim, Germany) the labeling of DNA strands with desoxiundina triphosphate labeled with fluorescein was visualized by confocal microscopy (wavelength excitation 488nm wavelength emission 525 nm) the laser intensity and the sensitivity of the photodetector were kept constant to allow comparison between treatments were analyzed at least 5 optical fields per animal

stainings TUNNEL were performed in subventπcular area rat brains, which continues to proliferate in the adult animal The administration of cannabinoids not only did not produce any significant apoptotic effects in the living m brain, but also the marking light observed in the caudate putamen of control animals was not observed in animals treated with cannabinoids

Claims

claims
1 - Use of natural and synthetic cannabinoids in the manufacture of a medicament for therapeutic treatment in mammals including humans, selected brain tumors from the group comprising glioblastoma, meduloepitehomas, medulloblastomas, neuroblastomas, germinoma, embryonal carcinomas, astrocytomas, astroblastomas, ependymomas, ohgodendrogliomas, plexales carcinomas, neuroepitheliomas, pineoblastomas epandimoblastomas, neuroectodermal tumors, malignant meningiomas, chondrosarcomas, meningeal sarcomatosomas, malignant melanomas and malignant schwanomas
2 - Use according to claim 1, wherein the brain tumors are ghoblastomas
3 - Use according to claims 1 and 2, in which the natural cannabinoids are selected from the group consisting of Δ 9 -tetrahιdrocannabιnol (THC), Δ 8 -tetrahιdrocannabιnol, cannabinol and cannabidiol
4 - Use according to any one of the preceding claims, wherein the natural cannabinoid is Δ 9 -tetrahιdrocannabιnol (THC)
5 - Use according to claims 1 and 2, in which the synthetic cannabinoids are selected from the group consisting of WIN 55,212-2-HU-210, CP-
55,940 and CP-50.556 (levonantradol)
6 - Use according to claims 1 and 5, in which the synthetic cannabinoid is WIN-55,212-2 7 - Drug for the treatment in mammals including meduloepite omas, medulloblastomas humans, brain tumors selected from the group comprising glioblastoma, neuroblastomas germinoma, embryonal carcinomas, astrocytomas, astroblastomas ependymomas, oligodendroghomas, plexales carcinomas, neuroepitehomas pineoblastomas epandimobiastomas, neuroectodermal tumors malignant meningiomas, chondrosarcomas, sarcomatosomas meningeal malignant melanomas and malignant schwanomas, characterized in that it comprises, as active ingredient, a natural or synthetic cannabinoid, and a pharmaceutically acceptable excipient
8 - Medicament according to claim 7, wherein the natural cannabinoid is selected from the group consisting of Δ 9 -tetrahιdrocannabιnol (THC) Δ 8 -tetrahιdrocannabιnol, cannabinol and cannabidiol
9 - Medicament according to claim 7, wherein the synthetic cannabinotde is selected from the group consisting of WIN 55,212-2-HU-210, CP-55,940 and CP-50.556 (levonantradol)
10 - Medicament according to any one of claims 7 to 9 wherein the carrier is one suitable for intratumoral (intracraneal) administration or systemic, such as oral, intravenous or intrapentoneal
11 - Medicament according to claim 10, wherein the excipient for intratumoral administration is a buffered saline (PBS) supplemented with bovine serum albumin (BSA) and dialyzed deslipizada
12 - Medicament according to any one of claims 10 to 11, wherein the concentration of the cannabinoid in the liquid intratumoral administration is from 10 to 10000 mg / ml for the natural cannabinoid and 1 to 1000 .mu.g / ml for the synthetic cannabinoid
13 - Procedure for the therapeutic treatment in mammals including humans, selected brain tumors from the group comprising glioblastoma, meduloepitehomas, medulloblastomas, neuroblastomas, germinoma, embryonal carcinomas, astrocytomas, astroblastomas, ependymomas, oligodendrogliomas, plexales carcinomas, neuroepitheliomas, pineoblastomas epandimoblastomas neuroectodermal tumors, malignant meningiomas, chondrosarcomas, meningeal sarcomatosomas, malignant melanomas and malignant schwanomas, comprising administering to the animal affected by one of such tumors a therapeutically effective amount of a medicament as defined in any one of claims 7 to 12
14 - Process according to claim 13, wherein the administration is performed intratumorally
15 - Process according to claim 14, wherein the administered amount of cannabinoid (active ingredient) varies from 100 to 50,000 g for natural cannabinoids and 10 to 5000 mg for synthetic cannabinoids
PCT/ES2000/000450 2000-02-11 2000-11-22 Therapy with cannabinoids in the treatment of cerebral tumor WO2001058445A1 (en)

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ES00976087T ES2241670T3 (en) 2000-02-11 2000-11-22 Cannabinoid therapy for the treatment of brain tumors.
DE2000620111 DE60020111D1 (en) 2000-02-11 2000-11-22 Therapeutic application of cannabinoids on the brain tumor treatment
AT00976087T AT295162T (en) 2000-02-11 2000-11-22 Therapeutic application of cannabinoids on the brain tumor treatment
DE2000620111 DE60020111T2 (en) 2000-02-11 2000-11-22 Therapeutic application of cannabinoids on the brain tumor treatment
AU1397901A AU1397901A (en) 2000-02-11 2000-11-22 Therapy with cannabinoids in the treatment of cerebral tumor
EP20000976087 EP1177790B1 (en) 2000-02-11 2000-11-22 Therapy with cannabinoids in the treatment of cerebral tumor

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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1307188A1 (en) * 2000-05-17 2003-05-07 Atlantic Technology Ventures, Inc. Cannabinoid drugs
EP1307186A1 (en) * 2000-05-17 2003-05-07 Atlantic Technology Ventures, Inc. Methods for decreasing cell proliferation based on (3r, 4r)-delta8-tetrahydrocannabinol-11-oic acids
EP1307188A4 (en) * 2000-05-17 2005-07-06 Atlantic Technology Ventures I Cannabinoid drugs
EP1307186A4 (en) * 2000-05-17 2005-10-19 Atlantic Technology Ventures I Methods for decreasing cell proliferation based on (3r, 4r)-delta8-tetrahydrocannabinol-11-oic acids
US9084771B2 (en) 2007-05-17 2015-07-21 Sutter West Bay Hospitals Methods and compositions for treating cancer
GB2478072A (en) * 2008-06-04 2011-08-24 Gw Pharma Ltd THC and CBD for use in the treatment of brain tumours
GB2478072B (en) * 2008-06-04 2012-12-26 Gw Pharma Ltd Anti-tumoural effects of cannabinoid combinations
GB2478074B (en) * 2008-06-04 2012-12-26 Gw Pharma Ltd Anti-tumoural effects of cannabinoid combinations
US8632825B2 (en) 2008-06-04 2014-01-21 Gw Pharma Limited Anti-tumoural effects of cannabinoid combinations
GB2478074A (en) * 2008-06-04 2011-08-24 Gw Pharma Ltd THC and CBD for use in the treatment of tumours
US8790719B2 (en) 2010-03-12 2014-07-29 Gw Pharma Limited Phytocannabinoids in the treatment of cancer
US9675654B2 (en) 2010-03-12 2017-06-13 Gw Pharma Limited Phytocannabinoids in the treatment of cancer

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