WO2020198039A1 - Ketamine for treatment of sensory processing deficits - Google Patents

Abstract

Disclosed herein are methods of treatment of autism and sensory processing disorders and ADNP Syndrome.

Description

KETAMINE FOR TREATMENT OF SENSORY PROCESSING DEFICITS

[0001] This application claims the benefit of and priority under 35 U.S.C. § 119(e) to

U.S. Serial No.: 62/822,822 filed March 23, 2019, the contents of which is hereby incorporated by reference in its entirety.

[0002] This patent disclosure contains material that is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure as it appears in the U.S. Patent and Trademark Office patent file or records, but otherwise reserves any and all copyright rights.

[0003] All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. The disclosure of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described herein.

BACKGROUND OF THE INVENTION

[0004] The present invention provides novel compositions and methods for administering to individuals with ADNP syndrome or autism who exhibit disordered processing of sensory input, commonly referred to as sensory processing disorder. Sensory processing disorder can manifest as oversensitivity to sensory input, such as sound or touch, or an excessive or compulsive seeking of sensory input.

[0005] ADNP Syndrome

[0006] ADNP Syndrome (also known as Helsmoortel-VanDerAa Syndrome / HVDAS) is a rare genetic condition that causes a wide spectrum of signs and symptoms. Its hallmark features are intellectual disability and autism spectrum disorder, characterized by impaired communication and social interaction. Affected individuals also have distinctive fecial features and abnormalities of multiple body systems.

[0007] Individuals with ADNP syndrome have mild to severe intellectual disability and delayed development of speech and motor skills such as sitting and walking. They demonstrate many features typical for autism spectrum disorder, including, for example, repetitive behaviors and social interactions difficulties. ADNP syndrome is also associated with mood disorders or behavioral problems, such as anxiety, temper tantrums, attention-deficit/hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), as well as sleep problems. [0008] Many individuals with ADNP syndrome have distinctive facial features, which often include a prominent forehead, a high hairline, outside comers of the eyes that point upward or downward (upslanting or downslanting palpebral fissures), ptosis, a broad nasal bridge, and a thin upper lip. These individuals may also have unusually shaped ears or hand and finger abnormalities. Eye and vision abnormalities, such as strabismus and hyperopia, also occur in ADNP syndrome. Some people with ADNP syndrome have early eruption of primary teeth.

[0009] ADNP syndrome may also cause hypotonia and feeding difficulties in infancy. Some patients develop digestive system problems, such as gastroesophageal reflux, vomiting, and constipation. Other ADNP features include obesity, seizures, and heart abnormalities.

[0010] ADNP gene

[0011] ADNP syndrome is caused by mutations in the ADNP (Activity-Dependent Neuroprotector Homeobox) gene. The ADNP gene is located on the long arm of chromosome 20 (20ql3.13). The ADNP protein, encoded by the ADNP gene, is a homeodomain-containing zinc finger protein with transcription factor activity. It participates in the regulation of the activity and expression of other genes through chromatin remodeling and is thereby involved in many aspects of growth and development.

[0012] ADNP is particularly important for brain formation and normal brain development. Experimental data demonstrated that downregulation of ADNP by antisense oligonucleotides upregulated p53 and reduced the viability of intestinal cancer cells by 90%. It has been proposed that ADNP is involved in maintaining cell survival, possibly by modulating p53.

[0013] The ADNP protein was found to interact with proteins BRG1 (SMARCA4), BAF250A (ARID 1 A), and BAF170 (SMARCC2), all of which are components of the SWI/SNF chromatin remodeling complex. Domain analysis showed that the C-terminal domain of ADNP was required for its interaction with SWI/SNF proteins. Short hairpin RNAs that knocked down ADNP expression to 80%, but not to 50%, resulted in microtubule reorganization and changes in cell morphology, with reduced formation of cell processes and reduced cell number.

[0014] The ADNP protein was also shown to interact with the chromatin remodeler CHD4 and the chromatin architectural protein HPl to form a stable complex, referred to as ChAHP. Besides mediating complex assembly, ADNP recognizes DNA motifs that specify binding of ChAHP to euchromatin. Genetic ablation of ChAHP components in mouse embryonic stem cells resulted in spontaneous differentiation concomitant with premature activation of lineage-specific genes and in a failure to differentiate towards the neuronal lineage. The mechanism of ChAHP-mediated silencing is suggested to differ from canonical HP 1 -mediated silencing.

[0015] ADNP syndrome pathology

[0016] Initially via whole-exome sequencing, Helsmoortel et al. (2014) identified 9 different de novo heterozygous truncating mutations in the ADNP gene in 10 unrelated Helsmoortel-Van der Aa syndrome patients. All mutations occurred at the 3-prime end of the last exon of the ADNP gene and can result in the loss of at least the last 166 C- terminal residues of the ADNP protein, leading to its escape from nonsense-mediated inRNA decay. Helsmoortel noted that mutations in other SWI/SNF components of the BAF complex, such as SMARCB1 and ARID IB, have been identified in patients with intellectual disability, and hypothesized that the ADNP mutations cause a dominant- negative effect on the recruitment of the BAF complex, resulting in deregulation of gene expression and a disruption of neuronal processes.

[0017] In 4 patients with intellectual disability and varying syndromic features, the Deciphering Developmental Disorders Study (2015) identified 3 de novo heterozygous mutations in the ADNP gene.

[0018] The suspected general role of the ChAHP complex in governing cell fete plasticity may explain why ADNP mutations affect several organs and body functions and contribute to cancer progression. Ostapcuk et al. (2018) found that the integrity of the ChAHP complex is disrupted by nonsense mutations identified in patients with ADNP syndrome, and this could be rescued by aminoglycosides that suppress translation termination.

[0019] Autism Spectrum Disorder

[0020] Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by impairments in social communication as well as restricted and repetitive interests and behaviors. Within these broad symptom domains, the behavioral profiles of individuals with ASD are extremely heterogeneous. In recent years, disruptive gene mutations involving several hundred different genes have been identified as putative causes of ASD, yet they account for only 30% of ASD cases altogether. The ADNP gene is one of the most commonly affected single-genes in ASD. SUMMARY OF THE INVENTION

[0021] In one aspect, the expression of the Activity-Dependent NeuroProtector Homeobox gene (ADNP) in a mammal is increased by administering to the mammal an effective amount of Ketamine, when the mammal is in need of treatment for a condition related to insufficient activity of ADNP.

[0022] In some embodiments, the insufficient activity is related to insufficient expression of ADNP. In some embodiments, the insufficient activity is related to genetic alterations of the ADNP gene. In some embodiments, the genetic alterations include germ line or somatic mutations of the ADNP gene. In some embodiments, the genetic alterations include copy number variations of the ADNP gene.

[0023] In some embodiments, the mammal displays one or more symptoms including delayed speech and language development, impaired social interactions, neurological speech impairment, urinary incontinence, or abnormal temper tantrums. In some embodiments, the mammal displays autism spectrum disorder (ASD). In some embodiments, the condition includes abnormal sensory processing. In some embodiments, the condition is the ADNP Syndrome.

[0024] In some embodiments, ketamine is used as a reagent to increase the ADNP expression for laboratory research. In some embodiments, ketamine is provided to the cells of the mammal in vitro.

[0025] In some embodiments, the mammal is provided with another pharmaceutically active agent. In some embodiments, ketamine is administered in a pharmaceutical composition.

[0026] In some embodiments, the mammal is a human.

[002h In some embodiments, ketamine is administered orally, topically, or parenterally. In some embodiments, ketamine is administered intramuscularly, subcutaneously, or intravenously. In some embodiments, ketamine is administered according to a schedule. In some embodiments, ketamine is administered one or more times daily. In some embodiments, ketamine is administered one or more times weekly. In some embodiments, ketamine is administered as needed. In some embodiments, each dose of ketamine corresponds to the amount of between 0.1 mg/kg and 1.0 mg/kg. BRIEF DESCRIPTION OF THE FIGURES

[0028] To conform to the requirements for PCT patent applications, the figures presented herein are black and white representations of images originally created in color.

[0029] Figure 1 shows that sedative doses of Ketamine upregulates the expression of

ADNP.

[0030] Figure 2 shows that lower doses of Ketamine upregulates the expression of

ADNP.

DETAILED DESCRIPTION OF THE INVENTION

[0031] Ketamine is on the World Health Organization’s List of Essential Medicines for Children up to 12 years of age. Recent clinical evidence suggests that the use of general anesthetics on young children can promote learning deficits as they mature.

[0032] Ketamine is a dissociative anesthetic. It is a non-competitive N- methyl-d- aspartate (NMDA) antagonist that disrupts calcium homeostasis in neurons. It can be a neurotoxin in some animal models as well as a neuroprotectant in other animal models, and depending on the dosage, it has been demonstrated to be protective as well as destructive to certain brain regions.

[0033] It has been observed that anesthetic doses of ketamine induce gene expression of ADNP. Turner et al (2012) observed that ADNP gene expression increase significantly in the somatosensory cortex of post-natal day 7 rats following a sedative dose of ketamine.

[0034] For Figure 1, P7 rat pups were injected with vehicle (Veh), 20 mg/kg ketamine (K20; 4 doses over 3 hours) or ketamine plus NAP (5, 10, or 20 mg/kg; N5K20, N10K20, N20K20, respectively; NAP injected 15 min prior to ketamine). After 8h, animals were anesthetized, perfused with 4% PFA, brains removed and 60 pm coronal sections were processed for AC3-ir. AC3-positive cells in layers IV and V of the somatosensory cortex (SSCIV-V) were counted using non-biased stereology. Data expressed as means ± SE (N = 6 for all groups). Significant differences were observed between K20 and vehicle (p<0.001***) and between N10K20 vs K20 and N20K20 and K20 (p<0.01**; ANOVA; Bonferroni post-test). No significant difference in the means was found when N5K20 was compared to K20. Figure 1 clearly pointed to a significant upregulation of ADNP in P7 rat pup brain by anesthetic dose of Ketamine. [0035] Administration of low doses of ketamine have been observed to induce ADNP expression as well, including in the somatosensory cortex. Brown et al investigated the effect on ADNP of low doses of Ketamine. They found that low-dose ketamine administration increased ADNP in the FC and SSOI/PI. Cells positive for ADNP immunohistochemistry were counted using non-biased ste neology for ketamine and control animals. Importantly, in neither the FC nor the SSCll/HI did K5 and K20 differ significantly in ADNP (p>0.05)-positive cells, indicating that sub-anesthetic doses of ketamine are equally effective in up-regulating ADNP.

[0036] For Figure 2, Sub-anesthetic dose of ketamine was used to upregulate ADNP in the fasciola cinereum and somatosensory cortex (Il/III) in P7 rat. (A) and (B) Somatosensory cortex (layer Il/III) immunostained for ADNP (RB a-ADNP 1:000, Bethyl Laboratories, TX, Catalog #IHC-00118) following (A) saline or (B) 5 mg/kg ketamine pre-treatment followed by 20 mg/kg ketamine treatment. (C) Fasciola cinereum. Significant differences were found between groups (F (2, 24)=9.545, p<0.001). Sal-K5 (p<0.05) and Sal-K20 (p<0.001). (D) Somatosensory cortex (II/III). Significant differences were found between groups (F (2, 24)=7.296, p<0.01). Sal-K5 (p<0.05) and Sal-K20 ( p<0.01). Post-test comparisons were done using Tukey HSD post hoc analysis. Scale bar=501M.

[003h The somatosensory cortex has a significant role in the sensory- system in the human body, including a role in sensory processing. Individuals with an ADNP mutation who exhibit abnormal sensory behaviors due to autism and/or sensory processing disorder, may have reduced levels of ADNP protein in the somatosensory cortices. An increase in ADNP protein levels in the somatosensory cortex could cause or be associated with amelioration of sensory processing disorders and/or the symptoms associated with sensory processing disorders such as sensation-seeking or sensation-avoiding behaviors.

[0038] The inventor has invented and discloses herein a method of treating sensory processing disorder in individuals with autism and/or sensory processing disorder associated with ADNP mutation by administering a sub-anesthetic dose of ketamine.

[0039] In one aspect, the expression of the Activity-Dependent NeuroProtector Homeobox gene (ADNP) in a mammal is increased by administering to the mammal an effective amount of Ketamine, when the mammal is in need of treatment for a condition related to insufficient activity of ADNP.

[0040] In some embodiments, the insufficient activity is related to insufficient expression of ADNP. In some embodiments, the insufficient activity is related to genetic alterations of the ADNP gene. In some embodiments, the genetic alterations include germ line or somatic mutations of the ADNP gene. In some embodiments, the genetic alterations include copy number variations of the ADNP gene.

[0041] In some embodiments, the mammal displays one or more symptoms including delayed speech and language development, impaired social interactions, neurological speech impairment, urinary incontinence, or abnormal temper tantrums. In some embodiments, the mammal displays autism spectrum disorder (ASD). In some embodiments, the condition includes abnormal sensory processing. In some embodiments, the condition is the ADNP Syndrome.

[0042] In some embodiments, ketamine is used as a reagent to increase the ADNP expression for laboratory research. In some embodiments, ketamine is provided to the cells of the mammal in vitro.

[0043] In some embodiments, the mammal is provided with another pharmaceutically active agent. In some embodiments, ketamine is administered in a pharmaceutical composition.

[0044] In some embodiments, the mammal is a human.

[0045] In some embodiments, ketamine is administered orally, topically, or parenterally. In some embodiments, ketamine is administered intramuscularly, subcutaneously, or intravenously. In some embodiments, ketamine is administered according to a schedule. In some embodiments, ketamine is administered one or more times daily. In some embodiments, ketamine is administered one or more times weekly. In some embodiments, ketamine is administered as needed. In some embodiments, each dose of ketamine corresponds to the amount of between 0.1 mg/kg and 1.0 mg/kg.

[0046] Definitions

[0047] As used herein,“Mammal” includes, without limitation, mice; rats; rabbits; guinea pigs; dogs; cats; sheep; goats; cows; horses; primates, such as monkeys, chimpanzees, and apes, and, in particular, humans.

[0048] As used herein, "administration" or "administering" refers to a method of giving a dosage of a pharmaceutically active ingredient to a vertebrate.

[0049] As used herein, a “dosage” refers to an amount of therapeutic agent administered to a patient.

[0050] As used herein, a“daily dosage” refers to the total amount of therapeutic agent administered to a patient in a day. [0051] As used herein, the term ‘therapeutic agent” means a substance that is effective in the treatment of a disease or condition.

[0052] As used herein, “therapeutically effective amount” or“pharmaceutically effective amount” is meant an amount of therapeutic agent, which has a therapeutic effect. The dosages of a pharmaceutically active ingredient which are useful in treatment are therapeutically effective amounts. Thus, as used herein, a therapeutically effective amount means those amounts of therapeutic agent which produce the desired therapeutic effect as judged by clinical trial results and/or model animal studies.

[0053] As used herein, a“therapeutic effect” relieves, to some extent, one or more of the symptoms of a disease or disorder. For example, a therapeutic effect may be observed by a reduction of the subjective discomfort that is communicated by a subject (e.g., reduced discomfort noted in self-administered patient questionnaire).

[0054] The term “carrier” defines a chemical compound that facilitates the incorporation of a compound into cells or tissues. For example dimethyl sulfoxide (DMSO) is a commonly utilized carrier as it facilitates the uptake of many organic compounds into the cells or tissues of an organism.

[0055] The term“physiologically acceptable” defines a carrier or diluent that does not abrogate the biological activity and properties of the compound.

[0056] The pharmaceutical compositions described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed with other active ingredients, as in combination therapy, or suitable carriers or excipient(s). Techniques for formulation and administration of the compound or combination of compounds disclosed herein may be found in“Remington’s Pharmaceutical Sciences,” Mack Publishing Co., Easton, PA, 18th edition, 1990.

[005h Some embodiments provide the compound(s) or combination of compounds disclosed herein in tablets, film coated tablets, capsules, caplets, pills, gel caps, pellets, beads, or dragee dosage forms. Preferably, the formulations disclosed herein can provide favorable drug processing qualities, including, for example, but not limited to, rapid tablet press speeds, reduced compression force, reduced ejection forces, blend uniformity, content uniformity, uniform dispersal of color, accelerated disintegration time, rapid dissolution, low friability (preferable for downstream processing such as packaging, shipping, pick-and-pack, etc.) and dosage form physical characteristics (e.g., weight, hardness, thickness, friability) with little variation. [0058] The compound(s) or combination of compounds disclosed herein can be formulated readily, for example, by combining the drug substance with any suitable pharmaceutically acceptable excipient(s) for example, but not limited to, binders, diluents, disintegrants, lubricants, fillers, carriers, coatings, glidants, flavors, color additives, and the like, as set forth below. Such compositions can be prepared for storage and for subsequent processing.

[0059] Excipients

[0060] Acceptable excipients for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Handbook of Pharmaceutical

Excipients, 5th edition (Raymond C Rowe, Paul J Sheskey and Sian C Owen, eds. 2005), and Remington: The Science and Practice of Pharmacy, 21st edition (Lippincott Williams & Wilkins, 2005), each of which is hereby incorporated in its entirety. The term“carrier” material or“excipient” herein can mean any substance, not itself a therapeutic agent, used as a carrier and/or diluent and/or adjuvant, or vehicle for delivery of a therapeutic agent to a subject or added to a pharmaceutical composition to improve its handling or storage properties or to permit or facilitate formation of a dose unit of the composition into a discrete article such as a capsule, tablet, film coated tablet, caplet, gel cap, pill, pellet, bead, and the like suitable for oral administration. Excipients can include, by way of illustration and not limitation, diluents, disintegrants, binding agents, wetting agents, polymers, lubricants, glidants, coatings, sweetens, solubilizing agents, substances added to mask or counteract a disagreeable taste or odor, flavors, colorants, fragrances, and substances added to improve appearance of the composition.

[0061] The compositions and formulations can include any other agents that provide improved transfer, delivery, tolerance, and the like. These compositions and formulations can include, for example, powders, pastes, jellies, waxes, oils, lipids, lipid (cationic or anionic) containing vesicles (such as LipofectinTM), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax.

[0062] Any of the foregoing mixtures can be appropriate in treatments and therapies in accordance with the disclosure herein, provided that the active ingredient in the formulation is not inactivated by the formulation and the formulation is physiologically compatible and tolerable with the route of administration. See also Baldrick P. “Pharmaceutical excipient development: the need for preclinical guidance.” Regul. Toxicol. Pharmacol. 32(2):210-8 (2000), Charman WN“Lipids, lipophilic drugs, and oral drug delivery-some emerging concepts.” J. Pharrn. Sci. 89(8):967-78 (2000), and the citations therein for additional information related to formulations, excipients and carriers well known to pharmaceutical chemists.

[0063] In some embodiments, one or more, or any combination of the listed excipients can be specifically included or excluded from the formulations and/or methods disclosed herein. As will be appreciated by those of skill in the art, the amounts of excipients will be determined by drug dosage and dosage form size.

[0064] Lubricants

[0065] In some embodiments, lubricants are employed in the manufacture of certain dosage forms. For example, a lubricant will often be employed when producing tablets. In some embodiments, a lubricant can be added just before the tableting step, and can be mixed with the formulation for a minimum period of time to obtain good dispersal. In some embodiments, one or more lubricants can be used. Examples of suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, zinc stearate, stearic acid, talc, glyceryl behenate, polyethylene glycol, polyethylene oxide polymers (for example, available under the registered trademarks of Carbowax^® for polyethylene glycol and Polyox^® for polyethylene oxide from Dow Chemical Company, Midland, Mich.), sodium lauryl sulfate, magnesium lauryl sulfate, sodium oleate, sodium stearyl fumarate, DL-leucine, colloidal silica, and others as known in the art. Typical lubricants are magnesium stearate, calcium stearate, zinc stearate and mixtures of magnesium stearate with sodium lauryl sulfate.

[0066] Color Additives

[0067] In some embodiments, color additives also can be included. The colorants can be used in amounts sufficient to distinguish dosage form strengths. Preferably, color additives approved for use in drugs (21 CFR 74, which is incorporated herein by reference in its entirety) are added to the commercial formulations to differentiate tablet strengths. The use of other pharmaceutically acceptable colorants and combinations thereof are encompassed by the current disclosure.

[0068] Binders

[0069] Binders can be used, for example, to impart cohesive qualities to a formulation, and thus ensure that the resulting dosage form remains intact after compaction. Suitable binder materials include, but are not limited to, microcrystalline cellulose, gelatin, sugars (including, for example, sucrose, glucose, dextrose and maltodextrin), polyethylene glycol, waxes, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, povidone, cellulosic polymers (including, for example, hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), methyl cellulose, hydroxyethyl cellulose, and the like), hydroxypropyl cellulose (HPC), and the like. Accordingly, in some embodiments, the formulations disclosed herein can include at least one binder to enhance the compressibility of the major excipient(s). In some embodiments, the binder(s) is(are) sprayed on from solution, e.g. wet granulation, to increase binding activity.

[0070] Disintegrants

[0071] In some embodiments, disintegrants are used, for example, to facilitate tablet disintegration after administration, and are generally starches, clays, celluloses, algins, gums, or crosslinked polymers. Suitable disintegrants include, but are not limited to, crosslinked polyvinylpyrrolidone (PVP-XL), sodium starch glycolate, alginic acid, mcthacrylic acid DYB, microcrystalline cellulose, crospovidone, polacriline potassium, sodium starch glycolate, starch, pregelatinized starch, croscarmellose sodium, and the like. If desired, the pharmaceutical formulation can also contain minor amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, for example, sodium acetate, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, sodium lauryl sulfate, dioctyl sodium sulfosuccinate, polyoxyethylene sorbitan fatty acid esters, etc. and the like.

[0072] Coatings

[0073] In some embodiments, the formulations can include a coating, for example, a film coating. Where film coatings are involved, coating preparations can include, for example, a film-forming polymer, a plasticizer, or the like. Also, the coatings can include pigments and/or opacifiers. Non-limiting examples of film-forming polymers include hydroxypropyl methylcellulose, hydroxypropyl cellulose, methylcellulose, polyvinyl pyrrolidine, and starches. Non-limiting examples of plasticizers include polyethylene glycol, tributyl citrate, dibutyl sebecate, castor oil, and acetylated monoglyceride. Furthermore, non-limiting examples of pigments and opacifiers include iron oxides of various colors, lake dyes of many colors, titanium dioxide, and the like.

[0074] Diluents

[0075] In some embodiments, diluents are used, and are generally selected from one or more of tire compounds sucrose, fructose, glucose, galactose, lactose, maltose, invert sugar, calcium carbonate, lactose, starch, microcrystalline cellulose, lactose monohydrate, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, a pharmaceutically acceptable polyol such as xylitol, sorbitol, maltitol, mannitol, isomalt and glycerol, polydextrose, starch, or the like, or any mixture thereof.

[0076] Surfactants

[0077] In some embodiments, surfactants are used. The use of surfactants as wetting agents in oral drag forms is described in the literature, for example in H. Sucker, P. Fuchs, P. Speiser, Pharmazeutische Technologic, 2nd edition, Thieme 1989, page 260. It is known from other papers, such as published in Advanced Drag Deliver)' Reviews (1997), 23, pages 163-183, that it is also possible to use surfactants, inter alia, to improve the permeation and bioavailability of pharmaceutical active compounds. Examples of surfactants include, but are not limited to, anionic surfactants, non-ionic surfactants, zwitterionic surfactants and a mixture thereof. Preferably, the surfactants is selected from the group consisting of poly(oxyethylene) sorbitan fatty acid ester, poly(oxyethylene) stearate, poly(oxyethylene) alkyl ether, polyglycolated glyceride, poly(oxyethylene) castor oil, sorbitan fatty acid ester, poloxamer, fatty acid salt, bile salt, alkyl sulfate, lecithin, mixed micelle of bile salt and lecithin, glucose ester vitamin E TPGS (D-a- tocopheryl polyethylene glycol 1000 succinate), sodium lauryl sulfate, and the like, and a mixture thereof.

[0078] Glidants

[0079] In some embodiments, glidants are used. Examples of glidants which may be used include, but are not limited to, colloidal silicon dioxide, magnesium trisilicate, powdered cellulose, starch, talc and calcium phosphate, or the like, and mixtures thereof.

[0080] Suitable routes of administration may, for example, include oral, rectal, transmucosal, topical, or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injections, as well as intrathecal, direct intraventricular, intraperitoneal, intranasal, or intraocular injections. The compound or combination of compounds disclosed herein can also be administered in sustained or controlled release dosage forms, including depot injections, osmotic pumps, pills, transdermal (including electrotransport) patches, and the like, for prolonged and/or timed, pulsed administration at a predetermined rate.

[0081] The pharmaceutical compositions of the present disclosure may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tabletting processes.

[0082] Pharmaceutical compositions for use in accordance with the present disclosure thus may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. Any of the well- known techniques, carriers, and excipients may be used as suitable and as understood in the art; e.g., in Remington’s Pharmaceutical Sciences, above.

[0083] Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, cysteine hydrochloride, and the like. In addition, if desired, the injectable pharmaceutical compositions may contain minor amounts of nontoxic auxiliary substances, such as wetting agents, pH buffering agents, and the like. Physiologically compatible buffers include, but are not limited to, Hanks’s solution, Ringer’s solution, or physiological saline buffer. If desired, absorption enhancing preparations (for example, liposomes), may be utilized.

[0084] For transmucosal administration, penetrants appropriate to the barrier to be permeated may be used in the formulation.

[0085] Pharmaceutical formulations for parenteral administration, e.g., by bolus injection or continuous infusion, include aqueous solutions of the active compounds or solids in water-soluble form. In some embodiments, formulations may comprise minocycline and a divalent or trivalent cation (e.g., Ca²⁺, Mg²⁺, Zn²⁺). In various embodiments, the molar ratio of cation to minocycline can be greater than 3:1, 4:1, or 5:1. In some embodiments, the molar ratio is above 5:1. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or other organic oils such as soybean, grapefruit or almond oils, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0086] For oral administration, the compound(s) or combination of compounds disclosed herein can be formulated readily by combining the active compound with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compound or combination of compounds disclosed herein to be formulated as tablets, film coated tablets, pills, dragees, capsules, liquids, gels, get caps, pellets, beads, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained by combining the active compound with solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses. In addition, stabilizers can be added. All formulations for oral administration should be in dosages suitable for such administration. In some embodiments, formulations of the compound(s) or combination of compounds disclosed herein with an acceptable immediate release dissolution profile and a robust, scalable method of manufacture are disclosed.

[008h Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration.

[0088] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0089] For administration by inhalation, the compound or combination of compounds disclosed herein is conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0090] Further disclosed herein are various pharmaceutical compositions well known in the pharmaceutical art for uses that include intraocular, intranasal, and intraauricular delivery. Suitable penetrants for these uses are generally known in the art. Pharmaceutical compositions for intraocular delivery include aqueous ophthalmic solutions of the active compounds in water-soluble form, such as eyedrops, or in gellan gum (Shedden et al., Clin. Ther., 23(3):440-50 (2001)) or hydrogels (Mayer et al., Ophthalmologica, 210(2): 101-3 (1996)); ophthalmic ointments; ophthalmic suspensions, such as microparticulates, drug-containing small polymeric particles that are suspended in a liquid carrier medium (Joshi, A., J. Ocul. Pharmacol., 10(l):29-45 (1994)), lipid-soluble formulations (Aim et al., Prog. Clin. Biol. Res., 312:447-58 (1989)), and microspheres (Mordenti, Toxicol. Sci., 52(1): 101-6 (1999)); and ocular inserts. All of the above- mentioned references are incorporated herein by reference in their entireties. Such suitable pharmaceutical formulations are most often and preferably formulated to be sterile, isotonic and buffered for stability and comfort. Pharmaceutical compositions far intranasal delivery may also include drops and sprays often prepared to simulate in many respects nasal secretions to ensure maintenance of normal ciliary action. As disclosed in Remington's Pharmaceutical Sciences, 18th Ed., Mack Publishing Co., Easton, PA (1990), which is incorporated herein by reference in its entirety, and well-known to those skilled in the art, suitable formulations are most often and preferably isotonic, slightly buffered to maintain a pH of 5.5 to 6.5, and most often and preferably include antimicrobial preservatives and appropriate drug stabilizers. Pharmaceutical formulations for intraauricular delivery include suspensions and ointments for topical application in the ear. Common solvents for such aural formulations include glycerin and water.

[0091] The compound(s) or combination of compounds disclosed herein may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

[0092] In addition to the formulations described previously, the compound or combination of compounds disclosed herein may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compound or combination of compounds disclosed herein may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

[0093] For hydrophobic compounds, a suitable pharmaceutical carrier may be a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. A common cosolvent system used is the VPD co-solvent system, which is a solution of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™, and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol. Naturally, the proportions of a co-solvent system may be varied considerably without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity nonpolar surfactants may be used instead of POLYSORBATE 80™; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

[0094] Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide also may be employed, although usually at the cost of greater toxicity. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.

[0095] Agents intended to be administered intracellularly may be administered using techniques well known to those of ordinary skill in the art. For example, such agents may be encapsulated into liposomes. All molecules present in an aqueous solution at the time of liposome formation are incorporated into the aqueous interior. The liposomal contents are both protected from the external micro-environment and, because liposomes fuse with cell membranes, are efficiently delivered into the cell cytoplasm. The liposome may be coated with a tissue-specific antibody. The liposomes will be targeted to and taken up selectively by the desired organ. Alternatively, small hydrophobic organic molecules may be directly administered intracellularly.

[0096] Additional therapeutic or diagnostic agents may be incorporated into the pharmaceutical compositions. Alternatively or additionally, pharmaceutical compositions may be combined with other compositions that contain other therapeutic or diagnostic agents.

[0097] Methods of Administration

[0098] The compound(s) or combination of compounds disclosed herein or pharmaceutical compositions may be administered to the patient by any suitable means. Non-limiting examples of methods of administration include, among others, (a) administration though oral pathways, which includes administration in capsule, tablet, granule, spray, syrup, or other such forms; (b) administration through non-oral pathways such as rectal, vaginal, intraurethral, intraocular, intranasal, or intraauricular, which includes administration as an aqueous suspension, an oily preparation or the like or as a drip, spray, suppository, salve, ointment or the like; (c) administration via injection, subcutaneously, intraperitoneally, intravenously, intramuscularly, intradermally, intraorbitally, intracapsularly, intraspinally, intrastemally, or the like, including infusion pump delivery; (d) administration locally such as by injection directly in the renal or cardiac area, e.g., by depot implantation; as well as (e) administration topically; as deemed appropriate by those of skill in the art for bringing the compound or combination of compounds disclosed herein into contact with living tissue.

[0099] Pharmaceutical compositions suitable for administration include compositions where the compound(s) or combination of compounds disclosed herein is contained in an amount effective to achieve its intended purpose. The therapeutically effective amount of the compound or combination of compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated, and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize. More specifically, a therapeutically effective amount means an amount of compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0100] As will be readily apparent to one skilled in the art, the useful in vivo dosage to be administered and the particular mode of administration will vary depending upon the age, weight and mammalian species treated, and the specific use for which the compound or combination of compounds disclosed herein are employed. The determination of effective dosage levels, that is the dosage levels necessary to achieve the desired result, can be accomplished by one skilled in the art using routine pharmacological methods. Typically, human clinical applications of products are commenced at lower dosage levels, with dosage level being increased until the desired effect is achieved. Alternatively, acceptable in vitro studies can be used to establish useful doses and routes of administration of the compositions identified by the present methods using established pharmacological methods.

[0101] The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage farms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in farm prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the farm of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drag Administration for prescription drugs, or the approved product insert. Compositions comprising the compound or combination of compounds disclosed herein formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0102] Exemplary compositions for oral administration include suspensions which may contain, for example, microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners or flavoring agents such as those known in the art; and immediate release tablets which may contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and/or lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants such as those known in the art. Molded tablets, compressed tablets or freeze -dried tablets are exemplary forms which may be used. Exemplary compositions include those formulating the compound or combination of compounds disclosed herein with first dissolving diluents such as mannitol, lactose, sucrose and/or cyclodextrins. Also included in such formulations may be high molecular weight excipients such as celluloses (avicel) or polyethylene glycols (PEG). Such formulations may also include an excipient to aid mucosal adhesion such as hydroxy propyl cellulose (HPC), hydroxy propyl methyl cellulose (HPMC), sodium carboxy methyl cellulose (SCMC), maleic anhydride copolymer (e.g., Gantrez), and agents to control release such as polyacrylic copolymer (e.g., Carbopol 934). Lubricants, glidants, flavors, coloring agents and stabilizers may also be added for ease of fabrication and use.

[0103] Exemplary compositions for nasal aerosol or inhalation administration include solutions in saline which may contain, for example, benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, and/or other solubilizing or dispersing agents such as those known in the art.

[0104] Exemplary compositions for parenteral administration include injectable solutions or suspensions which may contain, for example, suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution, an isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono- or diglycerides, and fatty' acids, including oleic acid. [0105] Exemplary compositions for rectal administration include suppositories which may contain, for example, a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures, but liquify and/or dissolve in the rectal cavity to release the drug.

[0106] Exemplary compositions for topical administration include a topical carrier such as Plastibase (mineral oil gelled with polyethylene). For example, the compound or combination of compounds disclosed herein may be administered topically to treat peripheral vascular diseases and as such may be formulated as a cream or ointment.

[0107] In some embodiments, the composition disclosed herein can comprise at least 0.1% (w/w), 0.2% (w/w), 0.3% (w/w), 0.4% (w/w), 0.5% (w/w), 0.6% (w/w), 0.7% (w/w), 0.8% (w/w), 0.9% (w/w), 1.0% (w/w), 1.1% (w/w), or 1.2% (wAv) of a preservative. In some embodiments, the topical composition disclosed herein can comprise 0.1% (w/w), 0.2% (w/w), 0.3% (w/w), 0.4% (w/w), 0.5% (w/w), 0.6% (w/w), 0.7% (w/w), 0.8% (w/w), 0.9% (w/w), 1.0% (wAv), 1.1% (w/w), 1.2% (w/w), 1.5% (w/w), 2% (w/w), 3% (w/w), 4% (w/w), 5% (w/w), 6% (w/w), 7% (w/w), 8% (w/w), 9% (w/w), 10% (w/w), 20% (w/w) or 30% (w/w) of a preservative or a range defined by any two of the preceding values. In some embodiments, the preservative can include one or more components, two or more components or three or more components.

[0108] In some embodiments, the composition disclosed herein can comprise at least 0.1% (w/w), 0.2% (w/w), 0.3% (w/w), 0.4% (w/w), 0.5% (w/w), 0.6% (w/w), 0.7% (w/w), 0.8% (w/w), 0.9% (w/w), 1.0% (w/w), 1.1% (w/w), or 1.2% (w/w) of a preservative including phenoxyethanol, propyl paraben, and methyl paraben. In some embodiments, the topical composition disclosed herein can comprise 0.1% (w/w), 0.2% (w/w), 0.3% (w/w), 0.4% (w/w), 0.5% (w/w), 0.6% (w/w), 0.7% (w/w), 0.8% (w/w), 0.9% (w/w), 1.0% (w/w), 1.1% (w/w), 1.2% (w/w), 1.5% (w/w), 2% (w/w), 3% (w/w), 4% (w/w), 5% (w/w), 6% (w/w), 7% (w/w), 8% (w/w), 9% (w/w), 10% (w/w), 20% (w/w) or 30% (w/w) of a preservative including phenoxyethanol, propyl paraben, and methyl paraben or a range defined by any two of the preceding values.

[0109] In some embodiments, the composition may include colorants, deodorants, fragrances, perfumes, anti-foaming agents, lubricants, natural moisturizing agents, skin conditioning agents, skin protectants, skin benefit agents, solvents, solubilizing agents, suspending agents, wetting agents, humectants, propellants, dyes, pigments, and combinations thereof. [0110] In some embodiments, the composition may include additional components added to enhance the odor, texture or color of the composition. For example, fragrances may be added to enhance odor. For example, emulsifiers or inert spheres may be added to enhance texture. For example, colorants may be added to enhance color.

[0111] In some embodiments, the composition may be applied to a body portion, such as a hand, foot, knee, elbow, and the like to treat pain and/or inflammation of the body- portion. The composition may be applied by any suitable means, such as rubbing, spraying, rolling, wiping, and the like, and massaged into the body portion to be treated.

[0112] In some embodiments, the minocycline as disclosed and described herein and/or topical compositions thereof can be used in combination therapy with at least one other agent. In some embodiments, the minocycline disclosed herein and/or topical composition thereof is administered concurrently with the administration of another agent, which may be part of the same topical composition as the compound of the present invention or a different composition. In other embodiments, a topical composition of the present invention is administered prior or subsequent to administration of another agent.

[0113] In some embodiments the compositions described herein are incorporated into a patch or film for transdermal drag delivery. In some embodiments, such patches further comprise a porous or resorbable film, an active pharmaceutical agent, and optionally a transdermal carrier or penetration enhancer. Exemplary transdermal carriers include dimethylsulfoxide;l-dodecylazacycloheptan-2-one or laurocapran; dimethylacetamide; dimethylformamide; lauric acid; myristic acid; capric acid; caprylic acid; oleic acid; diethylene glycol; tetraethylene glycol; terpenes; essential oils of eucalyptus, chenopodium and ylang-ylang; dimethyl isosorbide; Oxazolidinones such as 4- decyloxazolidin-2-one; 2-pyrrolidone; N-methyl-2-pyrrolidone; urea; EDTA; Sodium Glycolate; polysorbates; sodium deoxycholate; polyethylene glycol ;PLA/PLGA nanoparticles; polymer nanoparticles; block-copolymer nanoparticles, especially those comprising Pluronic®-type polyethylene oxide-block-polypropylene oxide copolymers; porous silica nanoparticles; metallic nanoparticles, especially those comprising gold, palladium, and iron; metal oxide nanoparticles, especially those comprising TiOa and AI2O3; short chain alcohols such as ethanol, propanol, and butanol; and oils such as mineral oil and coconut oil. In some embodiments the compositions described herein are incorporated into an adhesive for a transdermal patch. In some further embodiments, the compositions described herein are incorporated into a resorbable film. In some embodiments, the active pharmaceutical agent is contained within a separate reservoir layer. In some embodiments, the transderm al patch consists of a single layer. In some embodiments, the transdermal patch is constructed in multiple layers.

Claims

WHAT IS CLAIMED IS:

1. A method of increasing the expression of the Activity-Dependent NeuroProtector Homeobox gene (ADNP) in a mammal comprising administering to the mammal an effective amount of Ketamine, wherein the mammal is in need of treatment for a condition related to insufficient activity of ADNP.

2. The method of claim 1, wherein the insufficient activity is related to insufficient expression of ADNP.

3. The method of claim 1, wherein the insufficient activity is related to genetic alterations of the ADNP gene.

4. The method of claim 3, wherein the genetic alterations include germ line or somatic mutations of the ADNP gene.

5. The method of claim 3, wherein the genetic alterations include copy number variations of the ADNP gene.

6. The method of claim 1, wherein the mammal displays one or more symptoms selected from the group consisting of delayed speech and language development, impaired social interactions, neurological speech impairment, urinary incontinence, and abnormal temper tantrums.

7. The method of claim 1, wherein the mammal displays autism spectrum disorder

(ASD).

8. The method of claim 1, wherein the condition includes abnormal sensory processing.

9. The method of claim 1 , wherein the condition is ADNP Syndrome.

10. The method of claim 1, wherein ketamine is used as a reagent to increase the ADNP expression for laboratory research.

11. The method of claim 1, wherein ketamine is administered to the cells of the mammal in vitro.

12. The method of claim 1, further comprising administering to the mammal another pharmaceutically active agent.

13. The method of 1, wherein the ketamine is administered in a pharmaceutical composition.

14. The method of claim 1, wherein the mammal is human.

15. The method of claim 1, wherein ketamine is administered orally, topically, or parenterally.

16. The method of claim 15, wherein ketamine is administered intramuscularly, subcutaneously, or intravenously.

17. The method of claim 15, wherein ketamine is administered sublingually, transmucosally, or intranasally.

18. The method of claim 1, wherein ketamine is administered according to a schedule.

19. The method of claim 18, wherein ketamine is administered one or more times daily.

20. The method of claim 18, where ketamine is administered one or more times weekly.

21. The method of claim 1, wherein ketamine is administered as needed.

22. The method of claim 15, wherein each dose of ketamine corresponds to the amount of between 0.1 mg/kg and 1.0 mg/kg.