Patent ep2653163a1 – estrogenic components for use in the treatment of neurological disorders – google patents diffuse anoxic brain injury

• figure 1 post-operative body weights of rat pups. Post-operative body weights of rat pups that were injected intraperitoneally from day 4 to day 7 including after delivery either by vehicle (saline solution) (vehicle), 5 mg/kg E4 (E4 5 mg/kg) or 50 mg/kg E4 (E4 50 mg/kg) or not injected (sham). Mean ± SEM of body weights of 7 rat pups from the sham group, 11 rat pups of vehicle groups, 7 rat pups from the E4 5 mg/kg group and 5 rat pups from the E4 50 mg/kg group are shown.

• figure 2 brain weights of rat pups. Brain weights of rat pups that were injected intraperitoneally from day 4 to day 7 including after delivery either by vehicle (saline solution) (vehicle), 5 mg/kg E4 (5 mg/kg) or 50 mg/kg E4 (50 mg/kg) or not injected (sham).Diffuse anoxic brain injury

mean ± SEM of brain weights upon sacrifice at day 14 after delivery of 7 rat pups from the sham group, 11 rat pups of vehicle groups, 7 rat pups from the E4 5 mg/kg group and 5 rat pups from the E4 50 mg/kg group are shown. Scalebar: 2 mm.

• figure 3 hematoxylin-eosin staining of brain sections of the hippocampus region of rat pups. Brains of rat pups were removed upon sacrifice at day 14 after delivery and paraformaldehyde-fixed and paraffin-embedded samples were proceeded for sectioning at the hippocampus region and hematoxylin-eosin staining. Rat pups were either injected intraperitoneally from day 4 to day 7 including after delivery either by vehicle (saline solution) (vehicle), 5 mg/kg E4 (E4 5 mg/kg) or 50 mg/kg E4 (E4 50 mg/kg) or not injected (sham).Diffuse anoxic brain injury

• figure 4 intact cell counting in hematoxylin-eosin-stained brain sections of rat pups. Intact cells were counted in hippocampus in dentate gyrus zone (DG), subgranular zone (SGZ) and cornu ammonis (CA1, CA2/CA3) and in cortex on hematoxylin-eosin-stained brain sections of rat pups. Intact cells were counted at magnification 400x in 3 fields of the respective brain area and the average is expressed as the intact cell number per visual field. Mean ± SEM of intact cell number/ visual field weights of 7 rat pups from the sham group, 11 rat pups of vehicle groups, 7 rat pups from the E4 5 mg/kg group and 5 rat pups from the E4 50 mg/kg group are shown.


With the term neurodegenerative disease or disorder is generally meant a neurological disorder characterized by the progressive loss of structure or function of neurons, including death of neurons.Diffuse anoxic brain injury exemplary neurodegenerative diseases include, but are not limited to, alzheimer’s disease (AD), parkinson’s disease (PD), frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), huntington’s disease (HD) and other polyglutamine expansion diseases, pick’s disease, progressive supranuclear palsy, striatonigral degeneration, cortico-basal degeneration, olivopontocerebellar atrophy, leigh’s disease, infantile necrotizing encephalomyelopathy, hunter’s disease, mucopolysaccharidosis, various leukodystrophies (such as krabbe’s disease, pelizaeus-merzbacher disease and the like), amaurotic (familial) idiocy, kuf’s disease, spielmayer-vogt disease, tay sachs disease, batten disease, jansky-bielschowsky disease, reye’s disease, cerebral ataxia, chronic alcoholism, beriberi, hallervorden-spatz; syndrome, cerebellar degeneration, and the like.Diffuse anoxic brain injury preferably, the neurodegenerative disease is alzheimer’s disease (AD) or parkinson’s disease (PD).

Further exemplary neurological disorders that may be treated using the estrogenic components or pharmaceutical compositions taught herein may include but are not limited to: demyelinating autoimmune disorders, such as multiple sclerosis; neurological deficits caused by infection of inflammatory diseases, such as creutzfeldt-jacob disease or other slow virus infectious diseases of the central nervous system (CNS), AIDS encephalopathy, post-encephalitic parkinsonism, viral encephalitis, bacterial meningitis or other CNS effects of infectious diseases; cortical motor function disorders, such as spasticity, paresis, clones, or hyperreflexia; cortical sensory perception disorders, such as vestibular function disorders, balance and coordination disorders, dizziness, gait problems, dyslexia, clumsiness, development delay, audition discrimination and modulation disorders, delayed and mechanical speech disorders, vision problems, eye movement and coordination disorders, or sensory disturbance disorders; lower cranial nerve dysfunctions, such as lack of coordination between speech, swallowing or smooth articulation; bowel function disorders, such as gastro-esophageal sphincter control problems; abnormal urinary functioning, such as enuresis, bedwetting, or urinary bladder control disorders; respiratory dysfunctions, such as excessive snoring, obstructive or central apnea, or abnormal respiratory response to oxygen and carbon dioxide levels; sleep-disordered breathing, such as sleep apnea, muscular dysfunction, or sudden infant death; developmental disorders, such as chiari malformation; or congenital diseases, such as down’s syndrome, morquio’s syndrome, spondyloepiphysial dysplasia, achondroplasia, or osteogenesis; neurological behavioral disorders, such as attention deficit hyperactivity disorder, psychological problems, including anxiety, bipolar disorder, scizophrenia, or depression, autism spectrum disorders, including autism, asperger syndrome, and pervasive behavioral disorders-not otherwise specified; anatomic conditions, such as platybasia, retroflexed odontoid, basilar invagination, and foramen magnum stenosis; acquired bone-softening conditions, such as rickets, paget’s disease, or hyperparathyroidism; metabolic bone disorders; connective tissue disorders, including hypermobility connective tissue disorders, such as ehlers danlos syndrome; cervico-medullary syndrome; renal, metabolic, or endocrine syndromes; autonomic neural function disorders that cause abnormal blood flow to the skin, abnormal sexual response, GERDS, dyspraxia, idiopathic scoliosis, headaches, neck pain, back pain, head pain, encephalomyelopathy in the setting of trauma, neoplasm, positional orthostatic tachycardia, and bulbar findings.Diffuse anoxic brain injury

As used herein, carrier or excipient includes any and all solvents, diluents, buffers (such as, e.G., neutral buffered saline, phosphate buffered saline, or optionally tris-HCI, acetate or phosphate buffers), solubilisers (such as, e.G., tween 80, polysorbate 80), colloids, dispersion media, vehicles, fillers, chelating agents (such as, e.G., EDTA or glutathione), amino acids (such as, e.G., glycine), proteins, disintegrants, binders, lubricants, wetting agents, stabilisers, emulsifiers, sweeteners, colorants, flavourings, aromatisers, thickeners, agents for achieving a depot effect, coatings, antifungal agents, preservatives (such as, e.G., thimerosaltm, benzyl alcohol), antioxidants (such as, e.G., ascorbic acid, sodium metabisulfite), tonicity controlling agents, absorption delaying agents, adjuvants, bulking agents (such as, e.G., lactose, mannitol) and the like.Diffuse anoxic brain injury the use of such media and agents for formulating pharmaceutical compositions is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient(s), its use in the therapeutic compositions may be contemplated. Suitable pharmaceutical carriers are described inter alia in remington’s pharmaceutical sciences, 18th ed., mack publishing co., easton, PA (1990).

For example, the pharmaceutical composition as taught herein may be administered parenterally (such as by intravenous, intracerebral, intracerebroventricular, intramuscular, or subcutaneous injection, or intravenous infusion) in the form of a parenterally acceptable aqueous solution, which is pyrogen-free and has suitable ph, isotonicity and stability.Diffuse anoxic brain injury alternatively, the pharmaceutical composition as taught herein may be administered orally, for example in the form of pills, tablets, lacquered tablets, sugar-coated tablets, granules, hard and soft gelatin capsules, aqueous, alcoholic or oily solutions, syrups, emulsions or suspensions, or rectally, for example in the form of suppositories, percutaneous or topically (including ocular administration), for example in the form of ointments, tinctures, sprays or transdermal therapeutic systems (such as, e.G. A skin patch), or by inhalation in the form of nasal sprays or aerosol mixtures, or, for example, in the form of microcapsules, implants or rods.

For an oral administration form, the compositions of the present invention can be mixed with suitable additives, such as excipients, stabilizers, or inert diluents, and brought by means of the customary methods into the suitable administration forms, such as tablets, coated tablets, hard capsules, aqueous, alcoholic, or oily solutions.Diffuse anoxic brain injury examples of suitable inert carriers are gum arabic, magnesia, magnesium carbonate, potassium phosphate, lactose, glucose, or starch, in particular, corn starch. In this case, the preparation can be carried out both as dry and as moist granules. Suitable oily excipients or solvents are vegetable or animal oils, such as sunflower oil or cod liver oil. Suitable solvents for aqueous or alcoholic solutions are water, ethanol, sugar solutions, or mixtures thereof. Polyethylene glycols and polypropylene glycols are also useful as further auxiliaries for other administration forms. As immediate release tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate, and lactose and/or other excipients, binders, extenders, disintegrants, diluents, and lubricants known in the art.Diffuse anoxic brain injury

For subcutaneous or intravenous administration, the estrogenic component(s) described herein, if desired with the substances customary therefore such as solubilizers, emulsifiers, or further auxiliaries, are brought into solution, suspension, or emulsion. The active ingredient(s) can also be lyophilized and the lyophilizates obtained used, for example, for the production of injection or infusion preparations. Suitable solvents are, for example, water, physiological saline solution, or alcohols, e.G. Ethanol, propanol, glycerol, in addition also sugar solutions such as glucose or mannitol solutions, or alternatively mixtures of the various solvents mentioned. The injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents, or solvents, such as mannitol, 1,3-butanediol, water, ringer’s solution, or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.Diffuse anoxic brain injury

Some preferred, but non-limiting examples of such preparations include tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, cremes, lotions, soft and hard gelatin capsules, suppositories, drops, sterile injectable solutions and sterile packaged powders (which are usually reconstituted prior to use) for administration as a bolus and/or for continuous administration, which may be formulated with carriers, excipients, and diluents that are suitable per se for such formulations, such as lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, polyethylene glycol, cellulose, (sterile) water, methylcellulose, methyl- and propylhydroxybenzoates, talc, magnesium stearate, edible oils, vegetable oils and mineral oils or suitable mixtures thereof.Diffuse anoxic brain injury the formulations can optionally contain other pharmaceutically active substances (which may or may not lead to a synergistic effect with the compounds of the invention) and other substances that are commonly used in pharmaceutical formulations, such as lubricating agents, wetting agents, emulsifying, and suspending agents, dispersing agents, desintegrants, bulking agents, fillers, preserving agents, sweetening agents, flavoring agents, flow regulators, release agents, etc.

After delivery, newborn rat pups were assigned to one of the following 4 groups: sham group (neither vehicle nor estetrol (E4) applied), vehicle treated group (saline solution is used as a vehicle), E4 5 mg/kg/per day group and E4 50 mg/kg/per day group.Diffuse anoxic brain injury from day 4 to day 7 inclusively rat pups were injected intraperitoneally (IP) either by vehicle (vehicle group) or E4 (5mg/kg or 50 mg/kg in accordance to the group assignment) or not injected at all (sham group). At day 7, 30 minutes after last injection, animals from vehicle and E4 (5 mg/kg or 50 mg/kg) groups passed through surgery encompassing left common carotid artery double ligation and cut, followed by hypoxia produced by the inhalation of 11%-8% of oxygen balanced by nitrogen at decreased concentration for 20 minutes, followed by inhalation of 8% oxygen and 92% nitrogen at constant concentration for 35 minutes. The sham group went through similar procedures without left common carotid artery ligation and hypoxia.Diffuse anoxic brain injury all the manipulations were performed at 37°C. Rat pups recovered with their dams until being sacrificed at postnatal day 14.