Treatment and prevention of glioblastoma

Abstract

The present invention relates to the use of selective β2 adrenergic receptor antagonists for treating glioma. In particular, the present invention relates to the use of alkanolamine derivatives or pharmaceutically acceptable acid addition salts thereof for treating glioblastoma in patients.

Description

[Technical Field]

[0001] Technical field to which the invention belongs The present invention relates to the field of treatment and prevention of gliomas, more particularly to the treatment and prevention of glioblastomas. [Background technology]

[0002] Background of the Invention Glioma is a type of cancer that begins in the brain or spine. It is called glioma because it arises from glial cells and / or their precursors. The most common site for glioma is the brain. Gliomas are classified by cell type, grade, and location. Gliomas are named according to the specific cell type they most closely resemble. The main types of glioma are: Ependymomas, i.e., gliomas derived from ependymal cells, Astrocytomas, i.e., gliomas derived from astrocytes; glioblastoma multiforme (GBM) is the most common astrocytoma; Oligodendroglioma, i.e., glioma of oligodendrocyte origin, Mixed gliomas, such as oligoastrocytomas, which contain cells derived from different types of glia.

[0003] Gliomas are further classified according to their grade, which is determined by pathological evaluation of the tumor. Thus, gliomas can be distinguished into low-grade gliomas, which are well differentiated (not anaplastic), benign, and have a better prognosis for the patient, and high-grade gliomas, which are undifferentiated or anaplastic, malignant, and have a poor prognosis.

[0004] Of the many grading systems in use, the most common is the World Health Organization (WHO) grading system for astrocytomas.

[0005] Treatment for brain gliomas varies depending on location, cell type, and grade. Treatment is often a combined approach using surgery, radiation therapy, and chemotherapy. Radiation therapy is in the form of external beam radiation therapy or a stereotactic approach using radiosurgery. Spinal cord tumors can be treated with surgery and radiation. Temozolomide is a chemotherapy drug that can effectively cross the blood-brain barrier and is used in therapy. Despite these approaches, most patients with high-grade gliomas succumb to their disease. Novel therapeutic interventions directed at key targets are needed to improve outcomes in this patient population.

[0006] Glioblastoma multiforme (GBM, WHO grade IV) is an aggressive brain tumor that presents as one of two subtypes with distinct clinical histories and molecular profiles. Primary GBM presents as acutely aggressive disease, while secondary GBM subtypes develop from the indolent progression of low-grade disease (Dolecek TA et al. 2012. Neuro Oncol 14 (Suppl 5):v1-v49).

[0007] Malignant gliomas, including GBM, are by far the most common brain cancer in adults and one of the most challenging to treat. Despite invasive mono- and multimodal treatments, including surgery, chemotherapy, radiation, and small-molecule inhibitors, survival has remained stable over the past 30 years, with median survival times currently less than one year after diagnosis. The failure of conventional treatments is multifactorial, including the highly invasive nature of GBM, limitations in drug delivery across the blood-brain barrier and neuroparenchyma, and genetic heterogeneity leading to intrinsic resistance to available treatments and the emergence of invasive, resistant clones. Thus, novel treatment options are needed. Summary of the Invention

[0008] Brief description of the invention We found that ICI 118,551 reduced the viability of the glioblastoma cell line U-87 (Figure 1) and inhibited the formation and proliferation of gliospheres of a subpopulation of cancer stem cells derived from human glioblastoma under recognized culture procedures (Figure 2). In addition, we demonstrated that ICI 118,551 reduced the expression of stemness biomarkers and increased neural differentiation markers in glioblastoma (Figure 3), and delayed tumor progression in a mouse xenograft model (Figure 4).

[0009] Thus, the present invention relates to selective antagonists of β2-adrenergic receptors for use in the treatment and / or prevention of glioma. [Brief explanation of the drawings]

[0010] [Figure 1] Figure 1. ICI 118,551 is effective as an antitumor drug by reducing cell viability in glioblastoma cell lines, including stem cell subpopulations. A: The U-87 cell line expresses β-adrenergic receptor type 2, as shown by confocal microscopy using a monoclonal antibody against the human β2-adrenergic receptor (Abcam). B: ICI-118,551 reduces the viability of the human glioblastoma cell line U-87 when grown in liquid suspension or adherently as oncospheres (gliospheres, stem cells). Viability was quantified by luminescence using the ATP-Glo ​​kit from Promega. C. β2-adrenergic receptor expression in the human glioblastoma cell line U87 was confirmed by mRNA expression by quantitative PCR in three-dimensional culture as gliospheres (HUVECs as a positive control for β2-adrenergic receptor expression). [Figure 2]Figure 2. ICI 118,551 inhibits the formation and proliferation of gliospheres derived from the human glioblastoma cell line U-87. A: Photograph of gliosphere formation after 72 hours of drug treatment. U-87 cells were cultured in gliosphere formation medium and treated with vehicle (control, left) or ICI 118,551 and various concentrations of propranolol ranging from 10 to 100 μM. When cells were treated with β-blockers, a graded inhibition of gliosphere formation was observed, evident at concentrations above 10 μM. B: Limiting dilution assay (ELDA) experiments on oncosphere formation in the presence of various concentrations of ICI 118,551 and propranolol. These plots clearly show an increased dilution factor following treatment with ICI 118,551 and propranolol compared to the control. This is most likely a result of the sensitivity of U-87 stem cells to ICI 118,551 and propranolol. [Figure 3] Figure 3. ICI 118,551 and propranolol reduced the expression of stemness biomarkers but increased neuronal differentiation markers in the U-87 glioblastoma cell line. A: Treatment of U-87 with ICI-118,551 and propranolol for 48 h reduced the expression of stemness-related genes such as GD3 synthase, ALDH1, prominin (CD133), Sox2, and Nanog, while B: favored the expression of genes involved in neuronal differentiation (MAP2, GFAP, and nestin). Corresponding mRNA expression was measured by RT-qPCR. [Figure 4]Figure 4. ICI 118,551 and propranolol delay tumor progression of U-87 xenografts in NSG immunosuppressed mice. A. Xenograft using U-87 adherent cells: Mice (n = 30) were inoculated with 106 cells of the U-87 glioblastoma cell line as xenografts in the flanks. Tumor volume was measured every 3 days. In A, when tumors reached a volume of approximately 100 mm3, mice were divided into three groups (n = 9–10). One group received 10 mg / kg body weight of propranolol, another group received the same dose of ICI 118,551, and the third group received vehicle (DMSO) alone daily. The drugs were injected intraperitoneally. No adverse effects were observed. B. Xenograft using U-87 gliospheres: Mice were inoculated with 105 spheroids derived from the glioblastoma cell line U-87. Mice were divided into three groups of 9–10 mice and treated immediately after xenografting with either 3 mg / kg body weight propranolol, ICI 118,551, or vehicle alone for 5 consecutive days. Tumors were measured once they reached a sufficient size for measurement. [Figure 5] Figure 5. U87 cells were cultured in DMEM-10% FCS and incubated with increasing doses of ICI and propranolol for 48 hours. mRNA-21 expression is significantly reduced after incubation with ICI compared to propranolol (left). Furthermore, ICI increases the expression levels of mRNA-21 targets, such as PDCD4 pro-apoptotic agent 1 (center) and PTEN-a glioblastoma inhibitor (right). Specific Description of the Invention

[0011] Detailed Description of the Invention The present invention relates to selective antagonists of β2-adrenergic receptors for use in the treatment and / or prevention of glioma.

[0012] Alternatively, the present invention relates to the use of a selective antagonist of β2-adrenergic receptors in the manufacture of a medicament for the treatment and / or prevention of glioma.

[0013] Alternatively, the present invention relates to a method for treating and / or preventing glioma in a patient, comprising administering to said patient a therapeutically effective amount of a selective antagonist of β2-adrenergic receptors.

[0014] The term "β2-adrenergic receptor" or "β2AR," as used herein, refers to a class A G protein-coupled receptor (GPCR) that responds to diffusible hormones and neurotransmitters and is found primarily in smooth muscle. There are two main adrenergic receptors, α and β, with several subtypes: The α receptor has the subtype α1 (G q coupled receptor) and α2(G i There are receptors. β receptors are of three subtypes: β1, β2, and β3. All three are G s They bind to proteins that in turn bind to adenylate cyclase. Agonists binding to these receptors result in an increase in the intracellular concentration of the second messenger cAMP.

[0015] Agonists that bind to β2-adrenergic receptors result in smooth muscle relaxation.

[0016] The term "β2-adrenergic receptor antagonist," as used herein, refers to a compound that binds to the β2-adrenergic receptor and lacks substantial ability to activate the receptor itself. The term "β2-adrenergic receptor antagonist" includes both neutral antagonists and inverse agonists. A "neutral antagonist" is a compound that blocks the action of an agonist but has no effect on intrinsic or spontaneous receptor activity. An "inverse agonist" can both block the action of an agonist at the receptor and attenuate the constitutive activity of the receptor. The term "antagonist" also includes competitive antagonists, which are drugs that bind to the same site as the natural ligand; noncompetitive antagonists, which bind to a site on the receptor that is different from the natural ligand; reversible antagonists, which bind and unbind to the receptor at a rate determined by receptor-ligand kinetics; and irreversible antagonists, which bind permanently to the receptor by forming a covalent bond at the active site or by binding so tightly that the dissociation rate is effectively zero.

[0017] The term "selective β2-adrenergic receptor antagonist" as used herein refers to an antagonist that is more selective for β2-adrenergic receptors than β1-adrenergic receptors. In certain embodiments, the selective β2-adrenergic receptor antagonist exhibits at least 10-fold greater potency in binding to β2-adrenergic receptors than β1-adrenergic receptors, i.e., a β2 / β1 selectivity ratio of at least 10. More preferably, the selective β2 receptor antagonist has a β2 / β1 selectivity ratio of at least 50. Even more preferably, the selective β2 receptor antagonist has a β2 / β1 selectivity ratio of at least 123. The affinity of various active agents for β2-adrenergic receptors and β1-adrenergic receptors can be determined by evaluating tissues and / or cell subtypes that contain most β2 receptors (e.g., rabbit ciliary processes, rat liver, cat choroid plexus or lung), tissues that contain most β1 receptors (e.g., cat and guinea pig heart), and tissues that contain a mixture (e.g., guinea pig trachea). Methods for determining relative binding selectivity for these different types of tissue are comprehensively disclosed in O'Donnell and Wanstall, Naunyn-Schmiedeberg's Arch. Pharmaco., 308, 183-190 (1979); Nathanson, Science. 204, 843-844 (1979); Nathanson, Life Sciences, 26, 1793-1799 (1980); Minneman et al., Mol. Pharmacol., 15, 21-33 (1979a); and Minneman et al., Journal of Pharmacology and Experimental Therapeutics, 211, 502-508 (1979).

[0018] A significant number of compounds are known that have selective β2-adrenergic antagonist activity suitable for use in the present invention. In certain embodiments, the selective β2-adrenergic receptor antagonist is an alkanolamine derivative of Formula I: [ka] (In the formula, R 1 is an alkyl group of up to 6 carbon atoms branched at the α-carbon atom, R 2 is alkyl of up to 3 carbon atoms, R 3 is hydrogen, halogen, or alkyl of up to 3 carbon atoms, and n is 1 or 2) or a pharmaceutically acceptable acid addition salt thereof.

[0019] The term "alkyl group," as used herein, refers to acyclic straight and branched groups that can be derived from an alkane by removal of a hydrogen atom and have the formula --CnH2n+1.

[0020] The term "halogen," as used herein, refers to an atom selected from fluorine, chlorine, bromine and iodine.

[0021] R 1 can be, for example, isopropyl or t-butyl. In certain embodiments, R 1 is isopropyl.

[0022] R 2 can be, for example, methyl or ethyl. In certain embodiments, R 2 is methyl.

[0023] R 3 can be, for example, hydrogen, chlorine, bromine, methyl, or ethyl. In certain embodiments, R 3 is methyl.

[0024] In certain embodiments, n is 1.

[0025] In certain embodiments, R 1 is isopropyl, and R 2 is methyl. In certain embodiments, R 1is isopropyl and R is methyl. In certain embodiments, R 1 is isopropyl and n is 1. In certain embodiments, R 2 is methyl and R 3 is methyl. In certain embodiments, R 2 is methyl and n is 1. In certain embodiments, R 3 is methyl and n is 1.

[0026] In certain embodiments, R 1 is isopropyl, and R 2 and R 3 is methyl. In another particular embodiment, R 1 is isopropyl, and R 2 is methyl and n is 1. In another particular embodiment, R 1 is isopropyl, and R 3 is methyl and n is 1. In certain embodiments, R 2 and R 3 is methyl and n is 1.

[0027] In more specific embodiments, R 1 is isopropyl, and R 2 and / or R 3 is methyl and n is 1.

[0028] In an even more specific embodiment, the alkanolamine derivative has formula II: [ka]

[0029] This compound of formula II is also known as ICI 118,551, whose chemical name is erythro-D,L-1(methylidene-4-yloxy)-3-isopropylaminobutan-2-ol. ICI 118,551 has a β2 / β1 selectivity ratio of at least 123, as determined and reported in Life Sciences, 27,671 (1980) and Bilski et al., J. Cardiovasc.Pharmacol., 5, 430-437 (1983).

[0030] The alkanolamine derivatives of formula I contain two asymmetric carbon atoms, i.e., -CHOH- and -CHR2-, and are therefore believed to exist in four optically active forms: two racemic diastereoisomeric forms, the threo and erythro forms, and the (+) and (-) isomers of each racemic form. The present invention should be understood to encompass any one of these isomeric forms that possess selective β2-adrenergic receptor antagonist activity as defined above, and it is well known how to isolate a particular isomer and how to measure the selective β2-adrenergic receptor blocking activity it may possess.

[0031] It should be understood that, in general, optical isomers having the {S)-absolute configuration of the -CHOH- group are more active as β2 adrenergic blockers than the corresponding isomers having the {R)-absolute configuration. Generally, erythro isomers are more β2 selective than the corresponding threo isomers, although it is also known that both the threo and erythro isomers of the compounds of the invention possess the required selectivity.

[0032] The term "pharmaceutically acceptable acid addition salt" refers to an acid addition salt that can provide (directly or indirectly) a compound as described herein when administered to a recipient. Preferably, as used herein, the term "pharmaceutically acceptable salt" means that it is approved by a federal or state regulatory agency for use in animals, more particularly in humans, or is listed in the United States Pharmacopoeia or other generally recognized pharmacopeia. Salts can be prepared by methods known in the art. Illustrative non-limiting examples of pharmaceutically acceptable acid addition salts of the alkanolamine derivatives of Formula I include salts derived from inorganic acids, such as hydrochloride, hydrobromide, phosphate or sulfate, or salts derived from organic acids, such as oxalate, lactate, tartrate, acetate, salicylate, citrate, benzoate, β-naphthoate, adipate or 1,1-methylene-bis(2-hydroxy-3-naphthoate), or salts derived from acidic synthetic resins, such as sulfonated polystyrene resin.In certain embodiments, the pharmaceutically acceptable acid addition salt is hydrochloride.In more specific embodiments, the selective antagonist of β2-adrenergic receptor antagonist is the hydrochloride salt of the compound of Formula II.

[0033] In another particular embodiment, the selective β2-adrenergic receptor antagonist is selected from the list comprising the following compounds: Butoxamine corresponds to the compound with the chemical name DL-erythro-α-(2,5-dimethoxyphenyl)-β-t-butylaminopropanol hydrochloride. The determination of butoxamine's β2 selectivity was reported by O'Donnell and Wanstall, Naunyn-Schmiedeberg's Arch. Pharmaco., 308, 183-190 (1979), which reported a β2 / β1 selectivity ratio of at least 17. ·H35 / 25 corresponding to 1-(4'-methylphenyl)-b 2,2-l-isopropylaminopropanol. ·Structural formula: [ka] Prenalterol has the following selective β2-adrenergic receptor antagonist activity: This selective β2-adrenergic receptor antagonist activity has been described by Johansson and Waldeck, J. Pharm. Pharmacol., 1988, 32(9), 659-660. Various 4- and 5-[2-hydroxy-3-(isopropylamino)propoxy]benzimidazoles as described by Crooks et al., J. Med. Chem., 22(2), 210-214 (1979). 1-(t-butyl-amino-3-ol-2-propyl)oximino-9-fluorene as described by Imbs et al., Br. J. Pharmacol. 60(3), 357-362 (1977). Various 2-(α-hydroxyarylmethyl)-3,3-dimethylaziridines as described by Jain et al., J. Med. Chem., 21(1), 68-72 (1978).

[0034] The term "treatment," as used herein, refers to any method, action, application, therapy, etc. by which medical assistance is provided to a subject (or patient), including a human, for the purpose of directly or indirectly ameliorating the subject's condition, or slowing the progression of the subject's condition or disorder, or alleviating at least one symptom of the disease or disorder being treated.

[0035] The term "prevention," as used herein, refers to the administration of a compound of the present invention at an incipient or early stage of the disease, or to avoid its onset.

[0036] The term "glioma" as used herein refers to a general type of tumor originating from the brain. Gliomas originate from the glial cells that surround and support the neurons of the brain, including astrocytes, oligodendrocytes, and ependymal cells. Gliomas can be classified according to the specific cell types they share histological characteristics with (not necessarily their origin). The main types of gliomas are as follows: · Ependymoma: Ependymal cells. Astrocytoma: Astrocytes (glioblastoma multiforme is a malignant astrocytoma and is the most common primary brain tumor in adults). ·Oligodendroglioma: oligodendrocytes. Brainstem glioma: occurs in the brainstem. · Optic glioma: occurs in or around the optic nerve.

[0037] Mixed gliomas, such as oligoastrocytomas, contain cells of different glial origin.

[0038] The term "glioma" as used herein encompasses all types of gliomas, including ependymomas, astrocytomas, oligodendrogliomas, brainstem gliomas, optic gliomas, and oligoastrocytomas. In certain embodiments, gliomas comprise a subpopulation of cancer stem cells. As disclosed herein, cancer stem cells (CSCs) are a small subpopulation of cells within tumors that have the ability to self-renew, differentiate, and become tumorigenic. It is known in the art that various biomarkers, such as CD-44, Gremlin1, Id-1, TGFb2, BMP, OLIG2, SOX-2, ZEB1, Wnt5a, Pax-6, miRNSA-451, GD3S, and ALDH1, can be used to identify CSCs.

[0039] In certain embodiments, the glioma is an astrocytoma. The term "astrocytoma" includes grades I, II, III, and IV astrocytomas according to the WHO classification of tumors of the central nervous system. In more specific embodiments, the astrocytoma is a grade IV astrocytoma, also known as a "glioma" or "glioblastoma multiforme."

[0040] Gliomas can also be further classified according to their grade, which is determined by pathological evaluation of the tumor. Neuropathological evaluation and diagnosis of brain tumor specimens are performed according to the WHO classification of tumors of the central nervous system. Depending on their grade, they can be classified as follows: Low-grade gliomas (WHO grade II) are well differentiated (not anaplastic); they tend to be benign, meaning a better prognosis for the patient. High-grade (WHO grade III-IV) gliomas are undifferentiated or anaplastic, are malignant, and have a poor prognosis.

[0041] In certain embodiments, the glioma is a high-grade glioma.

[0042] Gliomas can be primary tumors or recurrent tumors. The terms "primary" or "non-recurrent" as used herein refer to tumors that appear for the first time in a subject, i.e., tumors that have not previously been detected and treated. The term "recurrent" as used herein refers to tumors that appear after a disease-free period, after treatment, and after a period in which cancer was not detected. In certain embodiments, the glioma is a primary or non-recurrent glioma.

[0043] In a preferred embodiment, the glioma is characterized by elevated expression of β2-adrenergic receptors compared to a reference value.

[0044] "Reference value" as used herein refers to a test value used as a reference for values / data obtained from a sample. A reference value (or reference level) can be an absolute value, a relative value, a value with upper and / or lower limits, a series of values, an average value, a median value, a mean value, or a value expressed by referring to a control or reference value. A reference value can be based on values ​​obtained from individual samples, such as values ​​obtained from test samples but at a previous time point. A reference value can also be based on multiple samples, such as values ​​obtained in a sample population or based on a sample pool that includes or excludes the test sample. In certain embodiments, the reference value is the expression of β2-adrenergic receptor in healthy subjects. In another specific embodiment, the reference value is the expression of β2-adrenergic receptor in subjects not affected by glioma.

[0045] The increase in β2-adrenergic receptor expression can be at least 2%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100% of the reference value.

[0046] Suitable methods for determining β2-adrenergic receptor expression include standard assays for determining mRNA expression levels, such as, but not limited to, qPCR, RT-PCR, RNA protection assays, Northern blots, RNA dot blots, in situ hybridization, microarray technology, tag-based methods such as serial analysis of gene expression (SAGE) (including variations such as LongSAGE and SuperSAGE), microarrays, and fluorescent in situ hybridization (FISH) (including variations such as Flow-FISH, qFISH, and double-fusion FISH (D-FISH)).

[0047] The term "patient" or "subject," as used herein, refers to an animal, preferably a mammal, including, but not limited to, domestic and farm animals, primates and humans, such as humans, non-human primates, cows, horses, pigs, sheep, goats, dogs, cats, or rodents. In preferred embodiments, the subject is a human of any age or race. In the present invention, the patient is suffering from glioma, more particularly astrocytoma, and even more particularly glioblastoma.

[0048] The term "glioma patient," as used herein, means that the patient has been diagnosed with glioma. A diagnosis of glioma can include: · Medical history and physical examination , including an interview about the patient's symptoms and personal and family health history. · Neurological examination The exam will check vision, hearing, speech, strength, sensation, balance, coordination, reflexes, and thinking and memory. It may also include an examination of the patient's eyes to look for swelling caused by pressure on the optic nerve, which connects the eye to the brain. · Brain scan Magnetic resonance imaging (MRI) and computed tomography (CT or CAT scan), which use a computer to produce detailed images of the brain, are the most common scans used to diagnose brain tumors. · biopsy This is a procedure to remove a small sample of the tumor for examination under a microscope. Depending on the location of the tumor, a biopsy and removal of the tumor may be done at the same time.

[0049] For its administration to a patient, the selective β2 adrenergic receptor antagonist of the present invention, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, can be formulated as a pharmaceutical composition.

[0050] The term "pharmaceutical composition," as used herein, refers to a composition comprising a therapeutically effective amount of a selective β2 adrenergic receptor antagonist of the present invention, preferably an alkanolamine derivative of formula I, more preferably an alkanolamine derivative of formula II, or a pharmaceutically acceptable acid addition salt thereof, and at least one pharmaceutically acceptable excipient or carrier.

[0051] The term "therapeutically effective amount," as used herein, refers to a sufficient amount of a compound to provide a desired effect, and is generally determined, among other things, by the cause, the characteristics of the compound itself, and the therapeutic effect to be achieved. It also varies depending on the subject being treated, the severity of the disease from which the subject suffers, the selected dosage form, the route of administration, etc. Therefore, the dosages set forth herein should be considered merely as a guide for those skilled in the art, who must adjust the dosage according to the above variables.

[0052] Although individual needs vary, determining the optimal range of therapeutically effective amounts of a compound for use according to the present invention is within the ordinary skill of one of ordinary skill in the art. Generally, the dosage required to provide effective treatment can be adjusted by one skilled in the art and will vary depending on the age, health, fitness, sex, diet, body weight, degree of receptor alteration, frequency of treatment, nature and condition of the injury, nature and extent of the disorder or disease, the health status of the subject, route of administration, pharmacological considerations such as the activity, efficacy, pharmacokinetics, and toxicology profile of the particular compound used, whether systemic drug delivery is used, and whether the compound is administered as part of a drug combination. The amount of a compound for use according to the present invention that is therapeutically effective in preventing and / or treating ischemia / reperfusion injury in a subject can be determined by conventional clinical techniques (see, e.g., The Physician's Desk Reference, Medical Economics Company, Inc., Oradell, NJ, 1995, and Drug Facts and Comparisons, Inc., St. Louis, MO, 1993).

[0053] In certain embodiments, a therapeutically effective amount results in the improvement of one or more symptoms of glioma. In certain embodiments, the selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered at a dose of about 0.2 mg / kg / day to about 5 mg / kg / day, preferably about 0.5 mg / kg / day, about 0.7 mg / kg / day, about 1 mg / kg / day, about 1.5 mg / kg / day, about 1.7 mg / kg / day, about 1.9 mg / kg / day, or about 2 mg / kg / day. In a more specific embodiment, the selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered at a dose of 2 mg / kg body weight / day to 3 mg / kg body weight / day. In an even more specific embodiment, the selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of formula I, more preferably an alkanolamine derivative of formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered at a dose of 2.4 mg / kg body weight / day.

[0054] The dose of the compound of the present invention can be expressed in mg of antagonist per kg of body weight or mg of antagonist per square meter of body surface. Those skilled in the art know how to determine the dose for a specific animal, particularly the dose for humans, from the dose experimentally tested in mice. For example, Reagan-Shaw S. et al. (Reagan-Shaw S. et al. "Dose translation from animal to human studies revisited". FASEB J 2008, 22(3):659-661) describes the conversion of mg / kg to mg / m 2 The standard transformation used to convert Dose (mg / kg) x K m =Dose(mg / m 2 )

[0055] The document also explains that this conversion is the basis for converting a dose in a first species to a dose in a second species (allometric dose conversion). Thus, an Animal Dose (AD) mg / kg can be converted to a Human Equivalent (HED) mg / kg using the following formula:

number

[0056] [Table 1]

[0057] Thus, studies using a dose of 30 mg / kg in mice correspond to a systemic human dose of 2.4 mg / kg.

[0058] In another specific embodiment, a selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered to a human at a fixed dose, each administration ranging from 0.2 mg / kg to 5 mg / kg, preferably about 0.2 mg / kg / day, about 0.25 mg / kg / day, about 0.3 mg / kg / day, about 0.35 mg / kg / day, about 0.4 mg / kg / day, about 0.45 mg / kg / day, about 0.50 mg / kg / day, about 0.55 mg / kg / day, about 0.6 mg / kg / day, about 0.65 mg / kg / day, about 0.7 mg / kg / day / day, about 0.75mg / kg / day, about 0.8mg / kg / day, about 0.85mg / kg / day, about 0.90mg / kg / day, about 0.95mg / kg / day, about 1mg / kg / day, about 1.2mg / kg / day, about 1.4mg / kg / day, about 1.6mg / kg / day, about 1.8mg / kg / day, about 2mg / kg / day, about 2.2mg / kg / day, about 2.4mg / kg / day, about 2.6mg / kg / day, about 2.8mg / kg / day, about 3mg / kg / day, about 3.2mg / kg / day to about 3.4mg / kg / day, about 3.6mg / kg / day, about 3.8mg / kg / day, about 4mg / kg / day, about 4.5mg / kg / day, about 5mg / kg / day. In a more preferred embodiment, the selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered at a dose of 2 mg / kg body weight / day and 3 mg / kg body weight / day. In an even more preferred embodiment, the selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered at a dose of 2.4 mg / kg body weight / day.

[0059] In another specific embodiment, the selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered daily, preferably once a day, twice a day, or three times a day. In a more preferred embodiment, it is administered once a day. In another specific embodiment, the selective β2 adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered for 2 days, 3 days, 4 days, 5 days, 7 days, 9 days, 10 days, 15 days, 20 days, 25 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more than 12 months, preferably for 5 days, even more preferably for 5 consecutive days.

[0060] The term "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to any compound or combination of compounds that is essentially non-toxic to a subject at the dosage and concentration used and is compatible with the other ingredients of a pharmaceutical composition. Thus, an excipient is an inert substance formulated with the active ingredient of a pharmaceutical composition (i.e., a selective β2-adrenergic receptor antagonist, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof) for the purpose of extending the composition containing the active ingredient. The extension allows for convenient and accurate dispensing of the drug substance when preparing a dosage form. Excipients may also serve various therapeutic purposes, such as aiding compound (drug) absorption or solubility or other pharmacokinetic considerations. Excipients may also be useful in manufacturing processes to aid in the handling of the active ingredient, such as by promoting powder flow or non-stickiness, as well as to aid in in vitro stability, such as preventing degradation during the expected storage period. The selection of suitable excipients depends on the route and dosage form of administration, as well as the active ingredient and other factors. Excipients can be any conventional non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating agent, or formulation aid. Illustrative, non-limiting examples of excipients or carriers include water, saline (physiological saline) solution, alcohol, dextrose, vegetable oil, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, flavor oils, mono- and diglycerides of fatty acids, petroleum fatty acid esters, hydroxymethylcellulose, and polyvinylpyrrolidone.

[0061] The selective β2 adrenergic receptor antagonist of the present invention, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, may be administered by any suitable route, including, but not limited to, parenteral, oral, topical, nasal, rectal, or intravitreal administration. In certain embodiments, the selective β2 adrenergic receptor antagonist of the present invention, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered intraperitoneally, intravenously, subcutaneously, intradermally, intramuscularly, or intravitreally. In a preferred embodiment, the selective β2 adrenergic receptor antagonist of the present invention, preferably an alkanolamine derivative of Formula I, more preferably an alkanolamine derivative of Formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered orally, intravenously, or intravitreally. In a more preferred embodiment, the selective β2 adrenergic receptor antagonist of the present invention, preferably an alkanolamine derivative of formula I, more preferably an alkanolamine derivative of formula II, or a pharmaceutically acceptable acid addition salt thereof, is administered intraperitoneally.

[0062] In a preferred embodiment, a selective β2-adrenergic receptor antagonist is administered together with an anti-tumor compound. As used herein, the term "tumor" or "cancer" refers to a broad group of diseases involving unregulated cell proliferation, also known as malignant neoplasms. This term is usually applied to diseases characterized by uncontrolled cell division (or enhanced survival or apoptosis resistance) and the ability of the cells to invade other adjacent tissues (invasion), spread via lymphatic and blood vessels to other areas of the body where they are not normally present (metastasis), circulate in the bloodstream, and then invade normal tissues elsewhere in the body. In certain embodiments, cancers are considered to be benign tumors, i.e., tumors that cannot spread by invasion or metastasis, i.e., they only grow locally. In another specific embodiment, cancers are considered to be malignant tumors, i.e., tumors that are capable of spreading by invasion and metastasis.

[0063] As used herein, the terms "antineoplastic agent," "anticancer agent," or "antineoplastic agent" (not used separately herein) generally refer to substances that inhibit or suppress the growth and proliferation of cancer cells. Antitumor agents can also include compounds that destroy cancer cells or prevent cell division, compounds that block certain hormones involved in cancer, compounds that inhibit or suppress the growth of new blood vessels (e.g., angiogenesis inhibitors), agents that damage DNA (e.g., alkylating agents such as cisplatin, carboplatin, and oxaloplatin; antimetabolites; and topoisomerase inhibitors), and compounds with anticancer properties (e.g., taxanes, vinca alkaloids, and plant alkaloids). The term "antineoplastic agent" also includes radiation therapy. Antitumor agents can also include agents specific to unregulated proteins in cancer cells. In a preferred embodiment, the antitumor compound is selected from leptomycin B (LMB, CAS No. 87081-35-4), temozolomide (CAS No. 85622-93-1), capecitabine (CAS No. 154361-50-9), and alkylating agents, intercalating agents, or DNA damaging agents. In a more preferred embodiment, the antitumor compound is propranolol (CAS No. 525-66-6).

[0064] As disclosed herein, an "alkylating agent" is a compound that adds an alkyl group to the guanine base of a DNA molecule, preventing the double helix strands from joining together as they should, resulting in DNA strand breaks and affecting the proliferation ability of cancer cells.

[0065] As disclosed herein, "intercalating agent" refers to a compound that inserts itself into the DNA structure of cells, binds to DNA, and causes DNA damage.In cancer treatment, DNA intercalating agents can kill cancer cells by damaging the DNA of cancer cells and stopping their division.

[0066] As disclosed herein, a "DNA damaging agent" is an agent that damages DNA by affecting the primary structure of the double helix, i.e., by chemically modifying the bases themselves. Non-limiting examples of damaging agents include reactive oxygen species, ultraviolet light, X-rays, and gamma rays.

[0067] The term "administered together with an anti-tumor compound" means that the selective β2-adrenergic receptor antagonist can be administered simultaneously or sequentially with the anti-tumor compound. When the selective β2-adrenergic receptor antagonist and the anti-tumor compound are administered simultaneously, they can be found in a single pharmaceutical composition or in different pharmaceutical compositions. When the selective β2-adrenergic receptor antagonist and the anti-tumor compound are administered sequentially, they can be administered in either order; that is, the administration of the selective β2-adrenergic receptor antagonist can begin before the administration of the anti-tumor compound, or the administration of the anti-tumor compound can begin before the administration of the selective β2-adrenergic receptor antagonist.

[0068] In a preferred embodiment, the selective β2-adrenergic receptor antagonist is administered to patients who, after first-line treatment, can no longer tolerate continuation of first-line treatment due to disease recurrence or severe secondary effects. In a preferred embodiment, the selective β2-adrenergic receptor antagonist is administered as second-line treatment. As disclosed herein, "second-line treatment" refers to treatment administered to patients who have previously received a first therapy known as "first-line treatment." First-line treatment is often part of a standard course of treatment, such as surgery followed by chemotherapy and radiation. It is also referred to as induction therapy, primary therapy, and primary treatment. As disclosed herein, any first-line treatment can be used. Non-limiting examples of first-line treatments for glioma include maximal surgical resection, individual radiation therapy, or radiation therapy with concomitant treatment and maintenance treatment of temozolomide (TMZ). Other examples of first-line treatments for glioma include anticancer compounds as defined above, such as DNA-damaging agents (e.g., cisplatin, carboplatin, and oxaloplatin); metabolic antagonists; topoisomerase inhibitors; and other compounds with anticancer properties (e.g., taxanes, vinca alkaloids, and plant alkaloids). In a preferred embodiment, the antitumor compound is selected from leptomycin B (LMB, CAS No. 87081-35-4), temozolomide (CAS No. 85622-93-1), capecitabine (CAS No. 154361-50-9), and alkylating agents, intercalating agents, or DNA-damaging agents. In a more preferred embodiment, the antitumor compound is propranolol (CAS No. 525-66-6).

[0069] In certain embodiments, the selective antagonist of β2-adrenergic receptors is administered as a first line therapy, ie, as a first treatment for glioma.

[0070] In a preferred embodiment, the selective β2-adrenergic receptor antagonist is administered during the disease-free period. As used herein, the term "disease-free period" refers to the period during and / or after treatment during which a patient survives with disease that has not progressed. In a preferred embodiment, the selective β2-adrenergic receptor antagonist is administered during the disease-free period that occurs after treatment with a first-line therapy, but the present invention also contemplates administering the selective β2-adrenergic receptor antagonist during other disease-free periods that a patient may undergo after a different line of therapy (after a second line of therapy, a third line of therapy, or an additional line of therapy). In another embodiment, the selective β2-adrenergic receptor antagonist is administered after the second line of therapy, after the third line of therapy, or after any line of therapy when the patient can no longer tolerate continuation of that line of therapy due to disease recurrence or excessively severe secondary effects.

[0071] The present invention is illustrated by the following examples, which should be considered as merely illustrative and not as limiting the scope of the invention. [Example]

[0072] material and method Oncosphere culture and extreme limiting dilution assay (ELDA) The protocol described elsewhere (Diaz-Guerra, E.; Lillo, MA; Santamaria, S.; Garcia-Sanz, JA, Intrinsic cues and hormones control mouse mammary epithelial tree size. FASEB J 2012, 26 (9), 3844-53) was followed. Cells were detached from plates with trypsin (Invitrogen) and seeded in complete medium (DMEM / F-12 medium containing GlutaMAX) supplemented with B27 (Gibco), 10 ng / ml epidermal growth factor (EGF; Invitrogen), and 10 ng / ml basic fibroblast growth factor (bFGF; Millipore), and maintained at 37°C in 5% CO2. For the ELDA assay, cells were dissociated with trypsin to form single cells and seeded at various dilutions (100 to 10 for U-87 cells) in complete sphere medium according to the procedure described by Ponti, D. et al. (Cancer Res 2005, 65 (13), 5506-11). The final number of spheres was quantified on day 14, and the final data and statistical significance were obtained using ELDA software (http: / / bioinf.wehi.edu.au / software / limdil / index.html and (Hu, Y. et al. J Immunol Methods 2009, 347 (1-2), 70-8).

[0073] RNA extraction and comparative quantification of miRNA and mRNA RNA was extracted from U-87 cells using the Direct-zol RNA MiniPrep kit (ZymoResearch). t RNA was isolated, washed with PBS, scraped off the plate, and spun down. The pellet was treated with Tri Reagent, homogenized at room temperature for 5 minutes, and then purified using the MiniPrep kit according to the manufacturer's instructions. tThe quality and concentration of RNA were assessed by measuring absorbance at 260, 230, and 280 nm using an ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). In all cases, the expected 260 / 280 (~2.0) and 260 / 230 (2.0-2.2) ratios, which are accepted as pure RNA, were obtained. To detect the two target miRs, 0.5 μg of RNA was extracted from cells. t was used for hybridization followed by measurements on a dual amperometric platform.

[0074] The expression of miR-21 and miR-205 was assessed using the qScript microRNA quantification system (Quanta BioSciences, Inc., Gaithersburg, MD, USA). Briefly, 10 ng of initial RNA was used per PCR reaction, and cDNA was synthesized using the qScript microRNA cDNA synthesis kit. PCR conditions consisted of an initial activation at 50°C for 2 minutes, followed by 40 cycles of 95°C for 5 seconds and 60°C for 30 seconds in a LightCycler 480 real-time PCR system (Roche). The C of each primer was t The threshold cycle values ​​were normalized to those of RNU6. To evaluate gene expression, cDNA was synthesized using a transcription product or first-strand cDNA synthesis kit (Roche) under the following PCR conditions: 95°C for 10 seconds, 60°C for 30 seconds, and 72°C for 30 seconds for 40 amplification cycles. ACTB was used as a reference gene for normalization, and real-time PCR reactions were performed in triplicate using PerfeCTa SYBR Green SuperMix (Quanta BioSciences) for miRNAs and FastStart Universal SYBR Green Master (Roche) for gene expression. The expression level of each gene was determined by the relative standard curve method, and primer sequences for the target genes were obtained from the Universal ProbeLibrary Assay Design Center (https: / / qpcr.probefinder.com / organism.jsp).

[0075] Confocal microscope Cells were seeded and grown in DMEM containing 10% FBS in P-24-well plates. After at least 24 hours of growth, cells were fixed in coverslips in 0.2% Triton X-100 / 3.5% PFA / PBS for 1 hour at 4°C. Cells were then permeabilized with PBS for 30 minutes at room temperature. Subsequently, cells were incubated with a monoclonal rabbit primary antibody against the β2 human adrenergic receptor (Abcam) at a 1:50 dilution in PBS for 60 minutes, washed three times with PBS, and incubated with Alexa-488 anti-rabbit secondary antibody at a 1:100 dilution in PBS for 1 hour. Coverslips were mounted using Prolong Gold with 4,6-diamidino-2-phenylindole (Invitrogen) and observed under a Leica TCS SP2 spectral confocal microscope (Leica Microsystems, Inc., Wetzlar, Germany).

[0076] Xenografts of U-87 glioblastoma cells and gliospheres A total of 10 injections were administered to the dorsal flank of 7-8 week-old male NOD scidγ (NSG) mice. 6 Cells from the U-87 cell line (A) or 10 cells obtained from dissociated gliospheres 5 In the case of A, the tumor size was 100 mm 3 When tumors reached a tumor volume of 1000 mg / kg, mice were randomly divided into three groups of 9 / 10 mice each. One group was treated with 10 mg / kg body weight of propranolol via daily intraperitoneal injection, the other group was treated with the same amount of ICI 118,551, and the control group was injected with vehicle. Tumor size was measured every 2–3 days with calipers. Mice were sacrificed when the mean tumor volume of the control group reached the endpoint established by our ethical procedures.

[0077] In case B, mice were divided into three groups of 10 and treated 5 days after xenograft establishment with an intraperitoneal injection of 3 mg / kg body weight of propranolol, ICI118,551, or vehicle. Mice were followed until tumors appeared and could be measured. From this point on, tumor size was measured every 2-3 days for 4 weeks, after which the mice were sacrificed.

[0078] statistical analysis Values ​​shown are the mean ± SEM or ± SD, and data from two groups were compared by t-test using GraphPad Prism 5. One-way analysis of variance (ANOVA) was used to determine mean differences for multiple pairwise comparisons, and p < 0.05 was considered significant.

[0079] result ICI 118,551 reduces viability and increases apoptosis in U-87 cells (human glioblastoma) in vitro Because ICI 118,551 is a selective β2-blocking agent, the first step to test was the expression of β2-adrenergic receptor type 2 in the U-87 cell line. As shown in Figure 1A, using confocal microscopy with a monoclonal antibody against human β2-adrenergic receptors (Abcam), we demonstrated that U-87 expresses this type of adrenergic receptor (Figure 1A, stained cells), thus demonstrating that this drug may exert its mechanism of action in this cellular model of glioblastoma. Expression of β2-adrenergic receptor mRNA was also confirmed by quantitative PCR in three-dimensional cultures as gliospheres (Figure 1C).

[0080] The mechanism of action of ICI 118,551 in U-87 cells was confirmed by treating the cells with various doses. It was clear that the drug reduced the viability of this human glioblastoma cell line model, as quantified by luminescence using the ATP-Glo ​​kit from Promega, when grown in liquid suspension as oncospheres (gliospheric stem cells) or adherently (Figure 1B). The results, shown in Figure 1B, demonstrate that ICI 118,551 differentially reduced U-87 cell viability when tested in liquid cultures of 3D gliospheres compared with 2D U-87 cells grown adherently. The differential sensitivity to the drug is shown in Figure 1B, with cells grown adherently being more sensitive than those grown in 3D gliospheres formed in suspension. While this is typical of most anticancer drugs, it is plausible that ICI 118,551 has a potent effect on the viability of cancer stem cells at low μM concentrations, which underlie the metastatic and recurrent processes of neoplastic disease.

[0081] ICI 118,551 inhibits the formation and proliferation of gliospheres in the human glioblastoma cell line U-87, a well-recognized culture procedure for cancer stem cells U-87 gliospheres were cultured under specific conditions as described elsewhere (Gupta, P.B. et al. 2009. Cell, 138, 645; Seymour, T. et al. 2015. Front. Oncol. 5:159. doi: 10.3389 / fonc.2015.00159; Penuelas et al. 2009. Cancer Cell, 15, 315-327; Lee, J. et al. 2006. Cancer Cell, 9, 391-403). As shown in Figure 2A, U-87 cells formed mature, spherical gliospheres with well-defined structures. U-87 cells were cultured in gliosphere-forming medium and treated with either vehicle (control, left) or different concentrations of ICI 118,551 and propranolol (10-100 μM). Treatment of cells with beta-blockers resulted in a dose-dependent inhibition of gliosphere formation, evident at concentrations above 10 μM for both propranolol and ICI-118,551. B: Limiting dilution assay (ELDA) of oncosphere formation in the presence of various concentrations of ICI-118,551 and propranolol. These plots clearly show an increased dilution factor after treatment with ICI-118,551 and propranolol compared to the control. This is a result of the stem cell sensitivity of U-87 stem cells to ICI-118,551 and propranolol.

[0082] ICI 118,551 and propranolol reduced mRNA expression of stemness biomarkers in U-87 glioblastoma cell line When U-87 cells were treated with ICI-118,551 and propranolol for 48 hours, the expression of stemness-related genes GD3 synthase, ALDH1, prominin (CD 133), Sox2, and nanog was significantly decreased (Figure 3A). However, mRNA expression from genes involved in neuronal differentiation, such as MAP2, GFAP, and nestin (Figure 3B), was decreased (Figure 3B). Corresponding mRNA expression was measured by RT-qPCR. We concluded that the expression of selected stemness biomarkers was decreased, while that of cell differentiation-related biomarkers was increased, all in a dose-dependent manner, collectively suggesting a role for ICI-118,551 in cell fate.

[0083] ICI 118,551 and propranolol delay tumor progression of U-87 xenografts in NSG immunosuppressed mice Xenografts were generated using U-87 adherent cells: 10 xenografts were placed in the flanks of mice (n=30). 6 The U-87 glioblastoma cell line was inoculated into the mice. Tumor volume was measured every 3 days. When tumors reached 100 mm 3 When tumor volumes reached the normal range, mice were divided into three groups (n = 9-10). One group was treated daily with 10 mg / kg body weight of propranolol, another group was treated with the same dose of ICI118,551, and the third group was treated with vehicle (DMSO) alone. The drugs were injected intraperitoneally. No adverse effects were observed. As shown in Figure 4A, a significant reduction of approximately 30% in tumor volume was observed in the groups of mice treated with 10 mg / kg body weight of propranolol or ICI118,551.

[0084] On the other hand, xenografts using U-87 gliospheres were also used. These gliospheres were densely packed with CSCs (cancer stem cells). 10 gliospheres derived from the glioblastoma cell line U-87 were injected into mice. 5Mice were inoculated with spheroids of 10 ...

[0085] ICI 118,551 induces a greater decrease than propranolol in mRNA-21 expression in U87 glioblastoma cell line The mechanisms underlying the effects of miR-21 on tumorigenesis remain unclear, particularly because only a few targets for this miR have been experimentally demonstrated. The oncomiR miR-21 is known to exert transcriptional control over PDCD4 in malignant cells, and its endogenous protein is upregulated 3.5-fold by miR-21 inhibition. PDCD4 expression is downregulated or lost in several tumor types, making it a promising molecular target for the treatment of several cancers (Frankel, LB, et al., J Biol Chem, 2008. 283(2): pp. 1026-33). Re-expression of miR-21 is associated with the acquisition of epithelial-mesenchymal transition in breast cancer and glioblastoma (Zhou Q, Liu J, Quan J, Liu W, Tan H, Li W. Cancer Sci. 2018 Sep; 109(9):2651-2659).

[0086] U87 cells were cultured in DMEM-10% FCS and incubated with increasing doses of ICI and propranolol for 48 hours. The expression of mRNA-21 was significantly reduced after incubation with ICI compared to propranolol (Figure 5, left). Furthermore, ICI increases the expression levels of mRNA-21 targets, such as PDCD4 pro-apoptotic drug 1 (center) and PTEN-a glioblastoma inhibitor (right).

Claims

1. β for use in the treatment and / or prevention of glioma ２ - a selective antagonist of adrenergic receptors, The antagonist has the formula: 【Chemical 1】 2. A selective antagonist of the β 2 -adrenergic receptor, comprising an alkanolamine derivative of the formula: or a pharmaceutically acceptable acid addition salt thereof.

2. The glioma has a β ２ 2. A selective antagonist of β 2 -adrenergic receptors for use according to claim 1, characterized in that expression of β 2 -adrenergic receptors is elevated.

3. 3. The selective antagonist of β 2 -adrenergic receptors for use according to claim 1 or 2, administered together with an antitumor compound selected from temozolomide, leptomycin B, capecitabine and an alkylating agent, an intercalating agent or a DNA damaging agent.

4. The selective antagonist of β 2 -adrenergic receptors for use according to claim 3, wherein said antitumor compound is propranolol.

5. A selective antagonist of β 2 -adrenergic receptors for use according to any one of claims 1 to 4, wherein the glioma comprises a subpopulation of cancer stem cells.

6. The selective β 2 -adrenergic receptor antagonist for use according to any one of claims 1 to 5, wherein said glioma is a high-grade glioma.

7. The selective β 2 -adrenergic receptor antagonist for use according to any one of claims 1 to 6, wherein said glioma is a non-recurrent glioma.

8. The selective β 2 -adrenergic receptor antagonist for use according to any one of claims 1 to 7, wherein said glioma is an astrocytoma.

9. The selective β 2 -adrenergic receptor antagonist for use according to claim 8, wherein the astrocytoma is glioblastoma.

10. The selective antagonist of β 2 -adrenergic receptors for use according to any one of claims 1 to 9, wherein said pharmaceutically acceptable acid addition salt is the hydrochloride salt.

11. The selective antagonist of β2-adrenergic receptors for use according to any one of claims 1 to 10, wherein the alkanolamine derivative or a pharmaceutically acceptable acid addition salt thereof is administered at a dose of 1 mg / kg body weight / day to 4.5 mg / kg body weight / day.

12. The selective antagonist of β 2 -adrenergic receptors for use according to claim 11, wherein the alkanolamine derivative or a pharmaceutically acceptable acid addition salt thereof is administered at a dose of 2 mg / kg body weight / day to 3 mg / kg body weight / day.

13. The selective antagonist of β 2 -adrenergic receptors for use according to claim 12, wherein the alkanolamine derivative or a pharmaceutically acceptable acid addition salt thereof is administered at a dose of 2.4 mg / kg body weight / day.

14. A selective antagonist of β 2 -adrenergic receptors for use according to any one of claims 1 to 13, administered to a patient as first line treatment or after first line treatment.

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