US 20170044244A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2017/0044244 A1 Frey et al. (43) Pub. Date: Feb. 16, 2017

(54) TREATMENT OF CENTRAL NERVOUS (60) Provisional application No. 61/769,673, filed on Feb. SYSTEM DSORDERS BY INTRANASAL 26, 2013, provisional application No. 61/862,814, ADMINISTRATION OF IMMUNOGLOBULIN filed on Aug. 6, 2013. G Publication Classification (71) Applicants: Baxalta Incorporated, Bannockburn, IL (US); Baxalta GmbH, Glattpark (51) Int. Cl. (Opfikon) (CH) C07K 6/8 (2006.01) A6II 47/8 (2006.01) (72) Inventors: William H. Frey, White Bear Lake, (52) U.S. Cl. MN (US); Leah Ranae Bresin CPC ...... C07K 16/18 (2013.01); A61K 47/183 Hanson, Vadnais Heights, MN (US); (2013.01); A6 IK 2039/543 (2013.01) Sharon Pokropinski, Schaumburg, IL (US); Francisco M. Rausa, Vernon (57) ABSTRACT Hills, IL (US) The present invention provides, among other aspects, meth (21) Appl. No.: 15/335,027 ods and compositions for treating a central nervous system (CNS) disorder by delivering a therapeutically effective (22) Filed: Oct. 26, 2016 amount of a composition of pooled human immunoglobulin G (IgG) to the brain via intranasal administration of the Related U.S. Application Data composition directly to the olfactory epithelium of the nasal (63) Continuation of application No. 14/189,981, filed on cavity. In particular, methods and compositions for treating Feb. 25, 2014. Alzheimer's disease are provided. Patent Application Publication Feb. 16, 2017 Sheet 1 of 28 US 2017/0044244 A1

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TREATMENT OF CENTRAL NERVOUS Arch Pathol LabMed., 117:132-144 (1993); Perl D. P. Neurol SYSTEM DSORDERS BY INTRANASAL Clin., 18:847-864 (2000)). The characteristic amyloid ADMINISTRATION OF IMMUNOGLOBULIN plaques, used to confirm Alzheimer's disease post-mortem, G are formed largely by deposition of a small amyloid-beta (AB) peptide derived from the amyloid precursor protein CROSS REFERENCES TO APPLICATIONS (APP). 0001. This application is a Continuation of U.S. patent 0007 To date, the U.S. Food and Drug Administration application Ser. No. 14/189,981, filed Feb. 25, 2014, which (FDA) has approved two types of medications for the claims priority to U.S. Provisional Patent Application Ser. management of Alzheimer's disease: cholinesterase inhibi No. 61/769,673 filed Feb. 26, 2013, and 61/862,814 filed tors, including donepezil (e.g., ARICEPTR), rivastigmine Aug. 6, 2013, the disclosures of which are hereby incorpo (e.g., EXELONR), galantamine (e.g., RAZADYNER), and rated herein by reference in their entireties for all purposes. tacrine (e.g., COGNEXR); and the NMDA-type glutamate receptor inhibitor memantine (marketed under a number of BACKGROUND OF THE INVENTION different brands). Although a cure for Alzheimer's disease has not been identified, these therapies serve to alleviate 0002 The central nervous system (CNS) is the process cognitive symptoms such as memory loss, confusion, and ing center for the nervous system. CNS disorders can affect loss of critical thinking abilities in Subjects diagnosed with the brain, the spinal cord, and nerve endings, resulting in age-related dementia (e.g., Alzheimer's disease). In all, it is neurological and/or psychiatric disorders. CNS disorders estimated that healthcare spending on Alzheimer's disease can be caused by genetic inheritance, trauma, infection, and related age-related dementias in 2012 will be $200 autoimmune disorders, structural defects, tumors, and billion in the United States alone (Factsheet, Alzheimer's stroke. Certain CNS disorders are characterized as neuro Association, March 2012). degenerative disease, many of which are inherited genetic diseases. Examples of neurodegenerative diseases include 0008. In addition to these approved therapies, several Huntington's disease, ALS, hereditary spastic hemiplegia, studies have Suggested that pooled intravenous immuno primary lateral sclerosis, spinal muscular atrophy, Kenne globulin (IVIG) is effective in slowing the progression of dy's disease, Alzheimer's disease, a polyglutamine repeat symptoms in Alzheimer's patients (Dodel R C et al., J. disease, or Parkinson's disease. Treatment of CNS disorders, Neurol Neurosurg Psychiatry, October 75(10): 1472-4 e.g., genetic diseases of the brain such as Parkinson's (2004); Magga J. et al., J Neuroinflammation, December 7; disease, Huntington's disease, and Alzheimer's disease, 7:90 (1997); Relkin N R et al., Neurobiol Aging, 30(11): remain an ongoing problem. 1728-36 (2008); Puli L. et al., J Neuroinflammation May 29; 0003) Alzheimer's disease is a common form of age 9:105 (2012)). related dementia that causes gradual loss of cognitive func 0009 Immune globulin products from human plasma tion, including memory and critical thinking abilities. were first used in 1952 to treat immune deficiency. Initially, Alzheimer's disease is diagnosed clinically by through a intramuscular or Subcutaneous administration of immuno finding of progressive memory loss and decrease in cogni globulin isotype G (IgG) isolated from plasma were the tive abilities. However, confirmation of Alzheimer's disease methods of choice. However, IgG products that could be does not occur until after death. administered intravenously, referred to as intravenous 0004 Alzheimer's disease is becoming more prevalent in immunoglobulin (IVIG), were later developed to allow for developed nations, where an increase in the population of the administration of larger amounts of IgG necessary for elder persons has occurred due in part to improved health effective treatment of various diseases. Usually, IVIG con care. While less than 1% of the population under the age of tains the pooled immunoglobulin G (IgG) immunoglobulins 60 is affected by Alzheimer's, it is estimated that 25% to from the plasma of multiple donors, e.g., more than a 33% of persons develop some form of Alzheimer's by the hundred or more than a thousand blood donors. These age of 85. As of 2012, 5.4 million Americans were diag purified IgG products are primarily used in treating three nosed with Alzheimer's. As life expectancy continues to main categories of medical conditions: (1) immune deficien increase worldwide, the prevalence of Alzheimer's and other cies: X-linked agammaglobulinemia, hypogammaglobuline age-related dementia should continue to grow as well. mia (primary immune deficiencies), and acquired compro 0005 Alzheimer's disease is typically classified as either mised immunity conditions (secondary immune “early onset,” referring to cases that begin to manifest at deficiencies), featuring low antibody levels; (2) inflamma between 30 and 60 years of age in affected individuals, and tory and autoimmune diseases; and (3) acute infections. the more common “late onset Alzheimer's, in which symp 0010 Specifically, many people with primary immuno toms first become apparent after the age of 60. Although deficiency disorders lack antibodies needed to resist infec only about 10% of all Alzheimer's cases are familial, early tion. In certain cases these deficiencies can be supplemented onset Alzheimer's disease has been linked to mutations in by the infusion of purified IgG, commonly through intrave the amyloid precursor protein (app), presenilin 1 (psen1), nous administration (i.e., IVIG therapy). Several primary and presenilin 2 (psen2) genes, while late onset Alzheimer's immunodeficiency disorders are commonly treated in the disease has been linked to mutations in the apolipoprotein E fashion, including X-linked agammaglobulinemia (XLA), (apoE) gene (Ertekin-Taner N., Neurol Clin., 25:611-667 Common Variable Immunodeficiency (CVID), Hyper-IgM (2007)). Syndrome (HIM), Severe Combined Immunodeficiency 0006. Histopathologically, this neurodegenerative dis (SCID), and some IgG subclass deficiencies (Blaese and ease is characterized by the formation of amyloid plaques, Winkelstein, J. Patient & Family Handbook for Primary neurofibrillary tangles, amyloid angiopathy, and granolo Immunodeficiency Diseases. Towson, Md.: Immune Defi vacuolar degeneration in the cerebral cortex (Mirra et al., ciency Foundation: 2007). US 2017/0044244 A1 Feb. 16, 2017

0011 While IgG treatment can be very effective for precipitation with alcohol. However, it was realized by managing primary immunodeficiency disorders, this therapy Oncley et al. (J. Am. Chem. Soc., 1949, 71(2): 541-550), is only a temporary replacement for antibodies that are not Deutsch et al. (J. Biol. Chem., 1946, 164, 109-118), and being produced in the body, rather than a cure for the Kistler and Nitschmann (Vox Sang., 1962, 7, 414-424), that disease. Accordingly, patients depend upon repeated doses particular protein precipitates (Fraction II and Fraction of IgG therapy, typically about once a month for life. This II+III) from the Cohn method could be used as a starting therapy places a great demand on the continued production material for the purification of highly enriched immuno of IgG compositions. However, unlike other biologics that globulin compositions. In order to achieve the higher purity are produced via in vitro expression of recombinant DNA and safety required for the intravenous administration of IgG vectors, IgG is fractionated from human blood and plasma compositions, several purification and polishing steps (e.g. donations. Thus, the level of commercially available IgG is adsorption in general or all different chromatographic tech limited by the available supply of blood and plasma dona niques, Cross-flow-filtration, etc.) have been added to IgG tions. manufacturing processes after the alcohol fractionation 0012 Several factors drive the demand for IgG, including steps. the acceptance of IgG treatments, the identification of addi 0017 Current IgG manufactures typically rely on either a tional indications for which IgG therapy is effective, and Cohn method 6 Fraction II+III precipitate or a Kistler increasing patient diagnosis and IgG prescription. Notably, Nitschmann precipitate A as the starting material for down the global demand for IgG more than quadrupled between stream processing. Both fractions are formed by a two step 1990 and 2009, and continues to increase at an annual rate process in which proteins such as fibrinogen and Factor XIII between about 7% and 10% (Robert P., Pharmaceutical are removed by an initial precipitation step (Fraction I Policy and Law, 11 (2009) 359-367). For example, the precipitation) performed at high pH (7.2) with low ethanol Australian National Blood Authority reported that the concentration (8-10% V/v), followed by a second precipita demand for IgG in Australia grew by 11.1% for the 2010 tion step in which IgG is precipitated from the Fraction I 2011 fiscal year (National Blood Authority Australia Annual supernatant at pH 6.8 with 20-25% (v/v) ethanol (Fraction Report 2010-2011). II+III) or at pH 5.85 with 19% ethanol (v/v) ethanol (pre 0013. It has been reported that in 2007, 26.5 million liters cipitate A), while albumin and a significant portion of A1PI of plasma were fractionated, generating 75.2 metric tons of remain in the Supernatant. IgG, with an average production yield of 2.8 grams per liter 0018. These methods, while laying the foundation for an (Robert P. supra). This same report estimated that global entire industry of plasma derived blood proteins, were IgG yields are expected to increase to about 3.43 grams per unable to provide IgG preparations having Sufficiently high liter by 2012. However, due to the continued growth in purity for the chronic treatment of several immune-related global demand for IgG, projected at between about 7% and diseases, including Kawasaki Syndrome, immune thrombo 14% annually between now and 2015, further improvement cytopenic purpura, and primary immune deficiencies, with of the overall IgG yield will be needed to meet global out an undue occurrence of serious side effects. As such, demand. One of the factors that may drive increased demand additional methodologies employing various techniques, for pooled human immunoglobulins (e.g., IVIG) over the Such as ion exchange chromatography, were developed to next decade is whether or not IgG is approved for the provide higher purity IgG formulations. Hoppe et al. treatment of Alzheimer's disease. It is estimated that if these (Munch Med Wochenschr 1967 (34): 1749-1752), Falksve treatments are approved by major regulatory agencies, an den (Swedish Patent No. 348942), and Falksveden and additional 5% increase in demand for IVIG will be seen Lundblad (Methods of Plasma Protein Fractionation 1980) (Robert P. Supra). were among the first to employ ion exchange chromatogra 0014. Due in part to the increasing global demand and phy for this purpose. fluctuations in the available Supply of immunoglobulin 0019. It is common practice to administer IgG by intra products, several countries, including Australia and Eng venous (IV) injection (Imbach et al., Lancet 1 (8232): 1228 land, have implemented demand management programs to 31 (1981)). Intravenous IgG (IVIG) may be administered protect Supplies of these products for the highest demand alone or in combination with other compositions. IVIG is patients during times of product shortages. Thus, the devel often administered over a 2 to 5 hour period, once a day for opment of methodologies that reduce the amount of pooled 2 to 7 days, with follow-up doses every 10 to 21 days or immunoglobulin G needed to treat various indications will every 3 to 4 weeks. Such an administration regime is time be critical as the increase in demand for pooled immuno consuming and inconvenient for many patients. This incon globulin begins to outpace the increase in global manufac venience may be aggravated in the case of Alzheimer's turing output. patients, who may have difficulty sitting quietly during the 00.15 Pooled human immunoglobulin G (IgG) is manu infusion period, and may have to rely on their caregiver to factured according to different processes depending upon the bring them to an infusion center or Supervise their infusion. specific manufacturer. However, the origin of most manu 0020 Systemic IVIG administration may cause adverse facturing processes is found in the fourth installment of a side effects. For example, IVIG may cause backache, head series of seminal papers published on the preparation and ache, migraine, joint or muscle pain, general feeling of properties of serum and plasma proteins, Cohn et al. (J. Am. discomfort, leg cramps, rash, pain at the injection site, hives, Chem. Soc., 1946, 68(3): 459-475). This paper first dizziness, unusual fatigue or tiredness or weakness, chills, described a method for the alcohol fractionation of plasma fever, Sweating, increased heart rate, increased blood pres proteins (method 6), which allows for the isolation of a Sure, cough, redness of the face, upset stomach, upper fraction enriched in IgG from human plasma. abdominal pain, and vomiting. Immediate adverse effects 0016. The Cohn procedures were initially developed to post-IVIG administration which have been observed include obtain albumin at relatively high (95%) purity by fractional headache, flushing, malaise, chest tightness, fever, chills, US 2017/0044244 A1 Feb. 16, 2017

myalgia, fatigue, dyspnea, back pain, nausea, Vomiting, covery 1:131-139). However, each of the above-described diarrhea, blood pressure changes, tachycardia, and anaphy approaches for increasing the delivery of therapeutics lactic reactions. Orbach et al., Clin. Rev. Allergy Immunol., through BBB to gain access to the CNS are limited. One 29(3): 173-84 (2005). such limitation is that the above-described approaches rely 0021. Furthermore, the adverse side effects may vary on systemic delivery systems, e.g., administration directly or based on the IVIG manufacturer. Most manufactures prepa indirectly to the blood stream, which results in non-specific rations contain between 90% and 99% purified IgG in delivery of the therapeutic agent to other parts in the body, combination with stabilizers and liquid(s) for reconstitution. increasing the chance of adverse side effects. Orange et al. 2006 (J. Allergy Clin. Immunol. 117(4 Suppl.): 0027 Intranasal administration of therapeutics has S525); Voet al. 2006 (Clin. J. Am. Soc. Nephrol. 1(4): 844; become an increasingly explored method for delivering Stiehm et al. 2006 (J. Pediatr. 148(1): 6). For example, some therapeutic agents to the brain because it circumvents the manufacturers use maltose as a stabilizer while others use BBB and is a localized, non-invasive method for delivery. Sucrose or amino acids. Furthermore, intranasal administration offers the advan 0022. The sodium and sugar content in IVIG, along with tages, over more traditional methods of delivery (e.g., intra varying amounts of IgA and additional chemicals used in the venous, Subcutaneous, oral transmucosal, oral or rectal IVIG production can affect the tolerability and efficacy of administration), of being simple to administer, providing the brand of IVIG in patients. Specifically, older patients rapid onset of action, and avoiding first-pass metabolism. often suffer from co-morbid conditions that increase the risk Unfortunately, intranasal administration has only been of IVIG adverse side effects. For example, subjects with shown effective for the transport of small molecules, and to renal disorders, vascular disorders, or diabetes also have a a certain extent smaller Fc fusion proteins, to the brain. The heightened risk of renal insufficiency and thrombotic events delivery of larger molecules. Such as intact antibodies, has following IVIG administration because IVIG compositions not yet been demonstrated. The difficulty in transporting are commonly hyper-viscous and contain high concentra larger proteins is believed to be due to the limited perme tions of Sugar and salt. ability of tight junctions present in the olfactory epithelia, 0023. IVIG also carries the risk of catheter-related infec which likely excludes globular molecules having a hydro tion, i.e., an infection where the catheter or needle enters a dynamic radius of more than 3.6 A (Stevenson B R, et al., subjects vein or skin. Examples of catheter-related infection Mol Cell Biochem., 1988 October: 83(2):129-45). are tenderness, warmth, irritation, drainage, redness, Swell (0028 U.S. Pat. No. 5,624,898 to Frey describes compo ing, and pain at the catheter site. Accordingly, alternate sitions and methods for transporting neurologic agents, modes of administration would be beneficial from the stand which promote nerve cell growth and Survival or augment point of time, convenience, and adverse side effects. the activity of functioning cells, to the brain by means of the 0024. In addition to adverse side effects of systemic olfactory neural pathway. The neurological agents of the administration of IVIG, penetration of IVIG across the 898 patent are transported to the brain by means of the blood-brain barrier has been shown to be unpredictable and nervous system, rather than the circulatory system, so that intraventricular or intrathecal IgG may be necessary. For potentially therapeutic agents that are unable to cross the example, Haley et al. administered IVIG in the treatment of blood-brain barrier may be delivered to damaged neurons in meningeal inflammation caused by West Nile virus encepha the brain. The compositions described in the 898 patent litis. Haley et al. found that penetration of IVIG was include a neurologic agent in combination with a pharma unpredictable and posited that intrathecal or intraventricular ceutical carrier and/or additive which promote the transfer of administration may be required. Haley et al. 2003 (Clin. Inf. the agent within the olfactory system. The 898 patent does Diseases 37: e88-90). not teach intranasal administration of pooled human immu 0025. It is difficult to target the CNS with IV adminis noglobulins. tration therapeutic compositions because of the blood-brain 0029 PCT publications WO 2006/091332 and WO 2009/ barrier (BBB). The BBB provides an efficient barrier, pre 058957, both by Bentz et al., describe methods for the venting and/or limiting access to the CNS of therapeutic delivery of therapeutic polypeptides to the brain by fusing compositions administered intravenously into the peripheral the polypeptide to an antibody or antibody fragment and circulation. Specifically, the BBB prevents diffusion of most administering the resulting fusion protein intranasally. therapeutic compositions, especially polar compositions, Although the examples of the 332 and 957 PCT publica into the brain from the circulating blood. tions Suggest that Fc-fusion “mimetibodies' may be admin 0026. At least three methods for increasing the passage of istered intranasally, neither publication demonstrates deliv molecules through the BBB have been developed. First, ery of larger, intact antibodies. In fact, the 957 PCT lipophilic compounds Such as lipid-soluble drugs and polar publication, published three years after the 332 PCT pub drugs encased in a lipid membrane have been developed. lication, states that “in published delivery studies to date, Lipophilic compounds with a molecular weight of less than intranasal delivery efficiency to the CNS has been very low 600 Da can diffuse through the BBB. Second, therapeutic and the delivery of large globular macromolecules, such as compounds can be bound to transporter molecules which antibodies and their fragments, has not been demonstrated.” can cross the BBB through a saturable transporter system. The 957 PCT publication purports to solve this problem Examples of Saturable transporter molecules are transferrin, through the use of a permeability enhancer (e.g., membrane insulin, IGF-1, and leptin. Third, therapeutic compounds can fluidizers, tight junction modulators, and medium chain cross the BBB by binding the therapeutic compounds to length fatty acids and salts and esters thereof, as described polycationic molecules such as positively-charged proteins below), which enhances intranasal administration to the that preferentially bind to the negatively-charged endothelial central nervous system. Neither PCT publication teaches surface of the BBB. Patridge et al. 2003 (Mol. Interv. 3(2): intranasal administration of pooled human immunoglobu 90-105); Patridge et al. 2002 (Nature Reviews-Drug Dis lins. US 2017/0044244 A1 Feb. 16, 2017

0030 PCT publication WO 2003/028668 by Barstow et includes intranasally administering the composition to a al., describes the treatment of immune-mediated diseases by nasal epithelium of the Subject associated with trigeminal alimentary administration (i.e., administration to the diges nerve endings. tive tract) of pooled immunoglobulins. Although the 668 0036. In another aspect, the disclosure provides a method PCT publication discloses nasal administration of a compo for treating a central nervous system (CNS) disorder in a sition of pooled immunoglobulins, it is in the context of Subject in need thereof, the method including delivering a delivering the composition to the digestive tract. The 668 therapeutically effective amount of a composition compris PCT publication does not teach the delivery of pooled ing pooled human immunoglobulin G (IgG) to the brain of human immunoglobulins to the brain via intranasal admin the subject, where delivering the composition to the brain istration. includes intranasally administering the composition to the 0031 PCT publication WO 2001/60420 by Adjei et al., upper third of the nasal cavity of the subject. describes aerosol formulations of therapeutic polypeptides, 0037. In another aspect, the disclosure provides a method including immunoglobulins, for pulmonary delivery. These for treating a central nervous system (CNS) disorder in a aerosolizable compositions are formulated Such that after Subject in need thereof, the method including delivering a oral or nasal inhalation, the therapeutic agent is effectively therapeutically effective amount of a composition compris delivered to the patient’s lungs. The 420 PCT publication ing pooled human immunoglobulin G (IgG) to the brain of does not teach the delivery of therapeutic agents to the brain the subject, where delivering the composition to the brain via intranasal administration. includes intranasally administering the composition to one 0032. Accordingly, there is a need in the art for methods or both maxillary sinus of the subject. of treating central nervous system disorders, such as 0038. In one embodiment of the methods described Alzheimer's disease, using pooled human immunoglobulin above, the CNS disorder is selected from the group consist G that provide specific targeting to the CNS (e.g., admin ing of a neurodegenerative disorder of the central nervous istration primarily to the brain), reduce systemic distribution system, a systemic atrophy primarily affecting the central of the pooled immunoglobulins, and lower the therapeuti nervous system, an extrapyramidal and movement disorder, cally effected dose needed for administration. a demyelinating disorder of the central nervous system, an episodic or paroxysmal disorder of the central nervous BRIEF SUMMARY OF INVENTION system, a paralytic syndrome of the central nervous system, 0033. The present disclosure provides solutions to these a nerve, nerve root, or plexus disorder of the central nervous and other problems by providing methods and compositions system, an organic mental disorder, a mental or behavioral for the treatment of central nervous system disorders via disorder caused by psychoactive Substance use, a schizo intranasal administration of pooled human immunoglobulin phrenia, Schizotypal, or delusional disorder, a mood (affec G. Advantageously, intranasal administration provides tive) disorder, neurotic, stress-related, or Somatoform disor directed delivery of pooled IgG to the brain, bypassing the der, a behavioral syndrome, an adult personality or behavior requirement that it pass through the blood brain barrier disorder, a psychological development disorder, and a child (BBB). As shown herein, intranasal administration allows onset behavioral or emotional disorder. the delivery of intact IgG to the brain. This results in greater 0039. In one embodiment of the methods described efficiency for the treatment and reduces the necessary IgG above, the CNS disorder is selected from the group consist dose that must be administered to achieve the desired effect. ing of Alzheimer's disease, Parkinson's disease, multiple As pooled human IgG is isolated from donated human Sclerosis, amyotrophic lateral Sclerosis (ALS), Huntington's plasma, pooled IgG is a limited resource. The reduction in disease, cerebral palsy, bipolar disorder, Schizophrenia, and the effective dose of IgG provided by the present disclosure Pediatric acute-onset neuropyschiatric syndrome (PANS). effectively increases the therapeutic potential provided by 0040. In one embodiment of the methods described the world's supply of pooled human IgG. Furthermore, as above, the CNS disorder is selected from the group consist demonstrated herein, intranasal administration of IgG nearly ing of Alzheimer's disease, Parkinson's disease, multiple eliminates the systemic exposure caused by intravenous sclerosis, Pediatric Autoimmune Neuropsychiatric Disor administration, improving the overall safety profile of the ders Associated with Streptococcal infections (PANDAS), treatment. Finally, it was Surprisingly found that IgG is and Pediatric acute-onset neuropySchiatric syndrome efficiently transported to the brain when intranasally admin (PANS). istered in the absence of permeability enhancers, Some of which have neurostimulatory effects themselves. 0041. In one embodiment of the methods described 0034. In one aspect, the disclosure provides a method for above, the CNS disorder is selected from the group consist treating a central nervous system (CNS) disorder in a subject ing of Alzheimer's disease, multiple Sclerosis, and Parkin in need thereof, the method including delivering a therapeu son's disease. tically effective amount of a composition comprising pooled 0042. In one embodiment of the methods described human immunoglobulin G (IgG) to the brain of the subject, above, the CNS disorder is Alzheimer's disease. where delivering the composition to the brain includes 0043. In one embodiment of the methods described intranasally administering the composition directly to the above, intranasal administration of the composition includes olfactory epithelium of the nasal cavity of the subject. the use of a non-invasive intranasal delivery device. 0035. In another aspect, the disclosure provides a method 0044. In one embodiment of the methods described for treating a central nervous system (CNS) disorder in a above, intranasal administration of the composition includes Subject in need thereof, the method including delivering a administration of a liquid drop of the composition directly therapeutically effective amount of a composition compris onto the nasal epithelium, the nasal epithelium of the subject ing pooled human immunoglobulin G (IgG) to the brain of associated with trigeminal nerve endings, or the upper third the subject, where delivering the composition to the brain of the nasal cavity of the subject. US 2017/0044244 A1 Feb. 16, 2017

0045. In one embodiment of the methods described embodiment of the methods provided above, the pH of the above, intranasal administration of the composition includes composition is from 4.0 to 7.5 In another embodiment of the directed administration of an aerosol of the composition to methods provided above, the pH of the composition is from the nasal epithelium, the nasal epithelium of the subject 6.0 to 7.5 associated with trigeminal nerve endings, or the upper third 0060. In one embodiment of the methods described of the nasal cavity of the subject. above, the method includes intranasally administering to the 0046. In one embodiment of the methods described subject a dose of from 0.08 mg to 100 mg pooled human IgG above, the aerosol of the composition is a liquid aerosol. per kg body weight of the Subject (mg IgG/kg). In a specific 0047. In one embodiment of the methods described embodiment of the methods provided above, the method above, the aerosol of the composition is a powder aerosol. includes intranasally administering to the Subject a dose of 0.048. In one embodiment of the methods described from 0.2 mg to 40 mg pooled human IgG per kg body weight above, at least 40% of the pooled human IgG administered of the Subject (mg IgG/kg). In a specific embodiment of the to the Subject contacts the nasal epithelium of the Subject, methods provided above, the method includes intranasally the olfactory epithelium of the nasal cavity of the subject, a administering to the Subject a dose of from 5 mg to 20 mg nasal epithelium of the Subject associated with trigeminal pooled human IgG per kg body weight of the Subject (mg nerve endings, the upper third of the nasal cavity of the IgG/kg). In a specific embodiment of the methods provided subject, or one or both maxillary sinus of the subject. above, the method includes intranasally administering to the 0049. In one embodiment of the methods described Subject a dose of from 5 mg to 10 mg pooled human IgG per above, at least 50% of the pooled human IgG administered kg body weight of the Subject (mg IgG/kg). In a specific to the subject contacts the olfactory epithelium of the nasal embodiment of the methods provided above, the method cavity of the subject, the nasal epithelium of the subject includes intranasally administering to the Subject a dose of associated with trigeminal nerve endings, the upper third of from 1 mg to 5 mg pooled human IgG per kg body weight the nasal cavity of the Subject, or one or both maxillary sinus of the Subject (mg IgG/kg). of the subject. 0061. In one embodiment of the methods described 0050. In one embodiment of the methods described above, the method includes intranasally administering to the above, at least 60% of the pooled human IgG administered subject a fixed dose of from 50 mg to 10 g pooled human to the subject contacts the olfactory epithelium of the nasal IgG. In a specific embodiment of the methods provided cavity of the subject, the nasal epithelium of the subject above, the method includes intranasally administering to the associated with trigeminal nerve endings, the upper third of subject a fixed dose of from 100 mg to 5.0 g pooled human the nasal cavity of the Subject, or one or both maxillary sinus IgG. In a specific embodiment of the methods provided of the subject. above, the method includes intranasally administering to the 0051. In one embodiment of the methods described subject a fixed dose of from 500 mg to 2.5g pooled human above, the pooled human IgG composition does not contain IgG. a permeability enhancer. 0062. In one embodiment of the methods described 0.052. In one embodiment of the methods described above, the method includes intranasally administering to the above, the pooled human IgG composition consists essen Subject a dose of pooled human IgG at least twice monthly. tially of pooled human IgG and an amino acid. In a specific embodiment of the methods described above, 0053. In one embodiment of the methods described the method includes intranasally administering to the Subject above, the amino acid is selected from the group consisting a dose of pooled human IgG at least once weekly. In a of glycine, histidine, and proline. In a specific embodiment specific embodiment of the methods described above, the of the methods provided above, the amino acid is glycine. method includes intranasally administering to the Subject a 0054. In one embodiment of the methods described dose of pooled human IgG at least twice weekly. In a specific above, the pooled human IgG composition is an aqueous embodiment of the methods described above, the method composition. includes intranasally administering to the Subject a dose of 0055. In one embodiment of the methods described pooled human IgG at least once daily. In a specific embodi above, the pooled human IgG composition includes from 10 ment of the methods described above, the method includes mg/mL to 250 mg/mL pooled human IgG and from 50 mM intranasally administering to the Subject a dose of pooled to 500 mM glycine. human IgG at least twice daily. 0056. In one embodiment of the methods described 0063. In one embodiment of the methods described above, the pH of the composition is from 4.0 to 6.0. In above, the pooled human IgG composition includes at least another embodiment of the methods provided above, the pH 0.1% anti-amyloid f IgG. of the composition is from 4.0 to 7.5. In another embodiment 0064. In one embodiment of the methods described of the methods provided above, the pH of the composition above, the method further includes administering a second is from 6.0 to 7.5. therapy for the CNS disorder to the subject in need thereof. 0057. In one embodiment of the methods described 0065. In one embodiment of the methods described above, the pooled human IgG composition is a dry powder above, the second therapy for the CNS disorder is a cholin composition. esterase inhibitor. In a specific embodiment of the methods 0.058. In one embodiment of the methods described described above, the cholinesterase inhibitor is selected above, the dry powder composition is prepared from an from the group consisting of donepezil (e.g., ARICEPTR), aqueous solution including from 10 mg/mL to 250 mg/mL rivastigmine (e.g., EXELONR), galantamine (e.g., RAZA pooled human IgG and from 50 mM to 500 mM glycine. DYNE(R), and tacrine (e.g., COGNEX(R). 0059. In one embodiment of the methods described 0066. In one embodiment of the methods described above, the dry powder composition is prepared from an above, the second therapy for the CNS disorder is an aqueous solution having a pH of from 4.0 to 6.0. In another inhibitor of NMDA-type glutamate receptor. In a specific US 2017/0044244 A1 Feb. 16, 2017

embodiment of the methods described above, the inhibitor 0076. In one embodiment of the uses described above, of NMDA-type glutamate receptor is memantine. intranasal administration of the composition includes 0067. In another aspect, the disclosure provides the use of directed administration of an aerosol of the composition to a composition comprising pooled human immunoglobulin G the nasal epithelium, the nasal epithelium of the subject (IgG) for the treatment of a central nervous system (CNS) associated with trigeminal nerve endings, or the upper third disorder in a subject in need thereof by intranasal adminis of the nasal cavity of the subject. tration. 0077. In one embodiment of the uses described above, 0068. In some embodiments of the uses described above, the aerosol of the composition is a liquid aerosol. intranasal administration includes administration to the 0078. In one embodiment of the uses described above, nasal epithelium of the subject. In other embodiments of the the aerosol of the composition is a powder aerosol. uses described above, intranasal administration comprises 0079. In one embodiment of the uses described above, at administration to the olfactory epithelium of the nasal cavity least 40% of the pooled human IgG administered to the of the subject. In other embodiments of the uses described Subject contacts the nasal epithelium of the Subject, the above, intranasal administration includes administration to a olfactory epithelium of the nasal cavity of the subject, a nasal epithelium of the Subject associated with trigeminal nasal epithelium of the Subject associated with trigeminal nerve endings. In other embodiments of the uses described nerve endings, the upper third of the nasal cavity of the above, intranasal administration includes administration to subject, or one or both maxillary sinus of the subject. the upper third of the nasal epithelium of the nasal cavity of 0080. In one embodiment of the uses described above, at the subject. In yet other embodiments, of the uses described least 50% of the pooled human IgG administered to the above, intranasal administration includes administration to Subject contacts the nasal epithelium of the Subject, the one or both maxillary sinus of the subject. olfactory epithelium of the nasal cavity of the subject, a 0069. In one embodiment of the uses described above, nasal epithelium of the Subject associated with trigeminal the CNS disorder is selected from the group consisting of a nerve endings, the upper third of the nasal cavity of the neurodegenerative disorder of the central nervous system, a subject, or one or both maxillary sinus of the subject. systemic atrophy primarily affecting the central nervous 0081. In one embodiment of the uses described above, at system, an extrapyramidal and movement disorder, a demy least 60% of the pooled human IgG administered to the elinating disorder of the central nervous system, an episodic Subject contacts the nasal epithelium of the Subject, the or paroxysmal disorder of the central nervous system, a olfactory epithelium of the nasal cavity of the subject, a paralytic syndrome of the central nervous system, a nerve, nasal epithelium of the subject associated with trigeminal nerve root, or plexus disorder of the central nervous system, nerve endings, the upper third of the nasal cavity of the an organic mental disorder, a mental or behavioral disorder subject, or one or both maxillary sinus of the subject. caused by psychoactive Substance use, a schizophrenia, 0082 In one embodiment of the uses described above, Schizotypal, or delusional disorder, a mood (affective) dis the pooled human IgG composition does not contain a order, neurotic, stress-related, or Somatoform disorder, a permeability enhancer. behavioral syndrome, an adult personality or behavior dis 0083. In one embodiment of the uses described above, order, a psychological development disorder, and a child the pooled human IgG composition consists essentially of onset behavioral or emotional disorder. pooled human IgG and an amino acid. 0070. In one embodiment of the uses described above, 0084. In one embodiment of the uses described above, the CNS disorder is selected from the group consisting of the amino acid is selected from the group consisting of Alzheimer's disease, Parkinson's disease, multiple Sclerosis, glycine, histidine, and proline. In a specific embodiment of amyotrophic lateral Sclerosis (ALS), Huntington's disease, the methods provided above, the amino acid is glycine. cerebral palsy, bipolar disorder, schizophrenia, and Pediatric 0085. In one embodiment of the uses described above, acute-onset neuropyschiatric syndrome (PANS). the pooled human IgG composition is an aqueous compo 0071. In one embodiment of the uses described above, sition. the CNS disorder is selected from the group consisting of 0086. In one embodiment of the uses described above, Alzheimer's disease, Parkinson's disease, multiple Sclerosis, the pooled human IgG composition includes from 10 mg/mL Pediatric Autoimmune Neuropsychiatric Disorders Associ to 250 mg/mL pooled human IgG and from 50 mM to 500 ated with Streptococcal infections (PANDAS), and Pediatric mM glycine. acute-onset neuropyschiatric syndrome (PANS). 0087. In one embodiment of the uses described above, 0072. In one embodiment of the uses described above, the pH of the composition is from 4.0 to 6.0. In another the CNS disorder is selected from the group consisting of embodiment of the uses described above, the pH of the Alzheimer's disease, multiple Sclerosis, and Parkinson's composition is from 4.0 to 7.5. In another embodiment of the disease. methods provided above, the pH of the composition is from 0073. In one embodiment of the uses described above, 6.0 to 7.5. the CNS disorder is Alzheimer's disease. 0088. In one embodiment of the uses described above, 0.074. In one embodiment of the uses described above, the pooled human IgG composition is a dry powder com intranasal administration of the composition includes the use position. of a non-invasive intranasal delivery device. 0089. In one embodiment of the uses described above, 0075. In one embodiment of the uses described above, the dry powder composition is prepared from an aqueous intranasal administration of the composition includes Solution including from 10 mg/mL to 250 mg/mL pooled administration of a liquid drop of the composition directly human IgG and from 50 mM to 500 mM glycine. onto the nasal epithelium, the nasal epithelium of the subject 0090. In one embodiment of the uses described above, associated with trigeminal nerve endings, or the upper third the dry powder composition is prepared from an aqueous of the nasal cavity of the subject. solution having a pH of from 4.0 to 6.0. In another embodi US 2017/0044244 A1 Feb. 16, 2017

ment of the uses described above, the pH of the composition BRIEF DESCRIPTION OF DRAWINGS is from 4.0 to 7.5 In another embodiment of the uses 0098. The patent or application file contains at least one described above, the pH of the composition is from 6.0 to 7.5 drawing executed in color. Copies of this patent or patent 0091. In one embodiment of the uses described above, application publication with color drawing(s) will be pro the use includes intranasally administering to the Subject a vided by the Office upon request and payment of the dose of from 0.08 mg to 100 mg pooled human IgG per kg necessary fee. body weight of the Subject (mg IgG/kg). In a specific 0099 FIGS. 1A-1F show brain slices from rats used to embodiment of the uses described above, the use includes assess the biodistribution of intranasally administered IgG in intranasally administering to the Subject a dose of from 0.2 Example 2. Six 2 mm slices (3 rostral to the optic chiasm and mg to 40 mg pooled human IgG per kg body weight of the 3 caudal) were acquired. Subject (mg IgG/kg). In a specific embodiment of the uses 0100 FIG. 1G illustrates a brain bisected along the described above, the use includes intranasally administering midline. The bisected brain is further dissected in to mid to the Subject a dose of from 5 mg to 20 mg pooled human brain, pons, medulla, and cerebellum for biodistribution IgG per kg body weight of the Subject (mg IgG/kg). In a analysis. specific embodiment of the uses described above, the use 0101 FIGS. 2A-2B illustrate results of average brain includes intranasally administering to the Subject a dose of tissue 'I IgG concentrations (nM)30 and 90 minutes after from 5 mg to 10 mg pooled human IgG per kg body weight intranasal administration of IgG. FIG. 2A illustrates results of the Subject (mg IgG/kg). In a specific embodiment of the of brain tissue 'I IgG concentrations (nM)30 (n=8) and 90 uses described above, the use includes intranasally admin (n-6) minutes after administration of a liquid protein IgG istering to the Subject a dose of from 1 mg to 5 mg pooled preparation, normalized to a 6.0 mg dose. FIG. 2B illustrates human IgG per kg body weight of the Subject (mg IgG/kg). results of brain tissue 'I IgG concentrations (nM) 30 0092. In one embodiment of the uses described above, (n=12) and 90 (n-6) minutes after administration of a solid the use includes intranasally administering to the Subject a microsphere IgG preparation, normalized to a 6.0 mg dose. fixed dose of from 50 mg to 10 g pooled human IgG. In a 01.02 FIGS. 3A-3E illustrate IHC data on cortical and specific embodiment of the uses provided above, the use hippocampal brain slices. Plaque content was determined for includes intranasally administering to the Subject a fixed 12 mice from each cohort (WT-Saline, WT-High, TG-Saline, dose of from 100 mg to 5.0 g pooled human IgG. In a TG-Low, and TG-High; shown left to right in the charts, specific embodiment of the uses provided above, the use respectively). FIG. 3A shows the percent area covered by includes intranasally administering to the subject a fixed beta-amyloid plaques. Four slides from the cortex of each dose of from 500 mg to 2.5 g pooled human IgG. mouse were analyzed using Image.J Software. The data is 0093. In one embodiment of the uses described above, distributed in the given range by plaque radius size (in the method includes intranasally administering to the Subject micrometers). Significant differences between transgenic a dose of pooled human IgG at least twice monthly. In a treatment groups are marked in the graph with the p-value. specific embodiment of the uses described above, the FIG. 3B shows the average number of beta-amyloid plaques. method includes intranasally administering to the Subject a Four slides from the cortex of each mouse were analyzed dose of pooled human IgG at least once weekly. In a specific using Image.J Software. The data is distributed in the given embodiment of the uses described above, the method range by plaque radius size (in micrometers). Significant includes intranasally administering to the Subject a dose of differences between transgenic treatment groups are marked pooled human IgG at least twice weekly. In a specific in the graph with the p-value. FIG. 3C shows the average embodiment of the uses described above, the method number of beta-amyloid plaques. Four slides from the hip includes intranasally administering to the Subject a dose of pocampus of each mouse were analyzed using Image.J pooled human IgG at least once daily. In a specific embodi Software. The data is distributed in the given range by plaque ment of the uses described above, the method includes radius size (in micrometers). FIG. 3D shows the percent area intranasally administering to the Subject a dose of pooled covered by beta-amyloid plaques. Four slides from the human IgG at least twice daily. hippocampus of each mouse were analyzed using Image.J 0094. In one embodiment of the uses described above, Software. The data is distributed in the given range by plaque the pooled human IgG composition includes at least 0.1% radius size (in micrometers). FIG. 3E shows immunofluo anti-amyloid B IgG. rescent staining of amyloid plaques in the hippocampus and cortex of aged TG2576 transgenic mice (field of view=5.3 0.095. In one embodiment of the uses described above, mm). For the immunofluorescent staining, mice were intra the method further includes administering a second therapy nasally administered saline, low dose IgG, or high dose IgG for the CNS disorder to the subject in need thereof. three times weekly over a period of 8 months. Each value is 0096. In one embodiment of the uses described above, reported as the mean value for the cohorts-tstandard error. the second therapy for the CNS disorder is a cholinesterase 0103 FIG. 4 illustrates a Kaplan-Meier curve for survival inhibitor. In a specific embodiment of the uses described rates of transgenic and wild-type mice administered IgG above, the cholinesterase inhibitor is selected from the group intranasally. These mice belong to a different cohort than the consisting of donepezil (e.g., ARICEPTR), rivastigmine mice used for plaque analysis in FIG. 3. (e.g., EXELONR), galantamine (e.g., RAZADYNE(R), or 0.104 FIG. 5 Illustrates the seven coronal brain slices tacrine (e.g., COGNEX(R). which were hemisected from intranasal 'I IgG treated rats 0097. In one embodiment of the uses described above, used to assess CNS delivery in Example 8. the second therapy for the CNS disorder is an inhibitor of 0105 FIGS. 6A-6B show comparative results of the NMDA-type glutamate receptor. In a specific embodiment intactness of IgG sprayed through a device designed for of the uses described above, the inhibitor of NMDA-type intranasal delivery with that of non-sprayed control. FIG.6A glutamate receptor is memantine. shows a Coomassie stained, non-reducing gel of sprayed and US 2017/0044244 A1 Feb. 16, 2017 non-sprayed (control). IgG. FIG. 6B shows a Western blot of order in a subject by intranasal delivery of a therapeutically a reducing gel probed with an anti-IgG antibody. effective amount of pooled human immunoglobulin G (IgG) 0106 FIG. 7 shows results demonstrating the highly directly to the epithelium of the nasal cavity of the subject. efficient olfactory epithelium targeting of IN device admin In a specific embodiment, pooled human IgG is adminis istration in rats. The upper panel shows the deposition of IN tered directly to the olfactory epithelium of the nasal cavity. IgG after device administration of 15 uL of 25% IVIG In some embodiments, pooled IgG is delivered to the upper solution spiked with 0.01% fluorescein tracer in a rat. The third of the nasal cavity, e.g., above the lower turbinates. In lower panel shows the deposition pattern after deposition of some embodiments, pooled IgG is delivered to the brain via 15uL of 25% IVIG solution spiked with 0.01% fluorescein the trigeminal nerve after intranasal administration to the tracer administered via nose drops. OB-olfactory bulb. nasal respiratory epithelium. In a specific embodiment, OE-olfactory epithelium, RE-respiratory epithelium, pooled IgG is delivered to the brain via the maxillary nerve NS=naris. after intranasal administration to the nasal respiratory epi 0107 FIGS. 8A-8C illustrate data showing a decrease of thelium. In other embodiments, pooled IgG is delivered to amyloid load in the low IgG and high IgG intranasal the brain after administration to the maxillary sinus. treatment groups. FIG. 8A shows the total amyloid area 0.115. In some embodiments, methods and compositions (plaque and vasculature). FIG. 8B shows the number (ii) of for the treatment of Alzheimer's disease, multiple sclerosis, amyloid deposits (plaque and vasculature). FIG. 8C shows and Parkinson's disease via intranasal administration of the total intensity of all amyloid deposits (i.e., the Sum pooled human IgG are provided herein. In other embodi Intensity). ments, the methods and compositions provided herein are 0108 FIGS. 9A-9C illustrate data showing a decrease in useful for the treatment of CNS disorder known to one of amyloid is a result of a decrease in plaque load. FIG. 9A skill in the art including, without limitation, a neurodegen shows the total amyloid area (plaque and vasculature). FIG. erative disorder of the central nervous system, a systemic 9B shows the number (ii) of amyloid deposits (plaque and atrophy primarily affecting the central nervous system, an vasculature). FIG. 9C shows the total intensity of all amy extrapyramidal and movement disorder, a demyelinating loid deposits (i.e., the Sum Intensity). disorder of the central nervous system, an episodic or 0109 FIGS. 10A-10C illustrate data showing that the paroxysmal disorder of the central nervous system, a para vascular component of the amyloid was found to increase lytic syndrome of the central nervous system, a nerve, nerve slightly when a decrease in amyloid as a result of a decrease root, or plexus disorder of the central nervous system, an in plaque load was observed (FIG. 9). FIG. 10A shows the organic mental disorder, a mental or behavioral disorder vascular amyloid area. FIG. 10B shows the number (ii) of caused by psychoactive Substance use, a schizophrenia, vascular deposits. FIG. 10C shows the total intensity of Schizotypal, or delusional disorder, a mood (affective) dis vascular deposits (i.e., the Sum Intensity). order, neurotic, stress-related, or somatoform disorder, a 0110 FIGS. 11A-11B illustrate data showing the relative behavioral syndrome, an adult personality or behavior dis proportions of vascular and plaque amyloid as it contributes order, a psychological development disorder, or a child onset to total amyloid. FIG. 11A shows the relative plaque con behavioral or emotional disorder. In some embodiments, the tribution to total amyloid. FIG. 11B shows the relative CNS disorder is selected from the group consisting of vascular contribution to total amyloid. Alzheimer's disease, Parkinson's disease, multiple Sclerosis, 0111 FIGS. 12A-12F show Congo Red stained sagittal amyotrophic lateral sclerosis (ALS), Huntington's disease, sections captured with confocal fluorescent microscopy. cerebral palsy, bipolar disorder, schizophrenia, or Pediatric FIG. 12A shows a Z-Stack max intensity projection image acute-onset neuropyschiatric syndrome (PANS). In some created from five individual images at 10x with a 512x512 embodiments, the CNS disorder is selected from the group resolution. FIGS. 12B-12F show single images created from consisting of Alzheimer's disease, Parkinson's disease, mul thirty of Z-Stacks projections as shown in FIG. 12A, encom tiple sclerosis, Pediatric Autoimmune Neuropsychiatric Dis passing the whole tissue section that were tiled (6x5, 5% orders Associated with Streptococcal infections (PANDAS), overlap). Representative images from the groups: Tg-Saline or Pediatric acute-onset neuropyschiatric syndrome (PANS). with Thresholding, Full-Resolution, Portion of the cortex 011.6 Advantageously, it is shown herein that intranasal and hippocampus (FIG. 12A); Tg-Low without Threshold administration of IgG increased weight and Survival time of ing (FIG. 12B); WT-Saline with thresholding (FIG. 12C); Alzheimer's disease mice models. For example, it is shown Tg-Saline with thresholding (FIG. 12D); Tg-Low with in Example 6 that intranasal administration of IgG, at either thresholding (FIG. 12E); and Tg-High with thresholding high (0.8 g/kg once every two weeks) or low (0.4 g/k once (FIG. 12F). every two weeks) doses, prolonged the lifespan of TG2576 0112 FIGS. 13 A-13B illustrate data for the average mice. This result shows that intranasal administration of IgG staining intensity for the Astrocyte marker GFAP (FIG. 13A) is capable of increasing lifespan in the Alzheimer's mouse and the microglial marker CD11b (FIG. 13B). 0113 FIG. 14 is an example image of amyloid (blue), model, indicating efficacy in Alzheimer's treatment. GFAP (green) and CD11b (red) staining from a Tg2576 0117 Moreover, intranasal administration of IgG signifi mouse brain that had been treated with a high dose of IN cantly reduced plaques in the cerebral cortex of in the IgG. CD11b staining was often observed surrounding the Alzheimer's mouse model. It is shown in Example 4 that amyloid plaques. treatment with pooled human IgG reduced the percent area covered by plaques in the Alzheimer's mouse model by DETAILED DESCRIPTION OF INVENTION about 25%, when administered intranasally at either low (0.4 mg/kg/2 wk; p=0.014) or high (0.8 mg/kg/2 wk; p=0.037) Introduction dosage. This is further indication of the efficacy of intranasal 0114. The present disclosure provides methods and com administration of IgG in the treatment of Alzheimer's dis positions for treating a central nervous system (CNS) dis CaSC. US 2017/0044244 A1 Feb. 16, 2017

0118. As further demonstrated herein, intranasal admin the brain, it is Surprising that intranasal administration of istration results in a much more discriminate delivery of pooled immunoglobulins provides the effective results pooled human IgG to the brain, as compared to intravenous shown herein. administration. For example, it is shown in Example 9 that I0122) Advantageously, intranasal delivery of the pooled intranasal administration of pooled human immunoglobulin human IgG composition disclosed herein can be accom G resulted in a 6-fold lower blood exposure as compared to plished by a non-invasive means, as compared to intrave intravenous administration. The lower system exposure of nous, Subcutaneous, and intramuscular administration, all of IgG provided by intranasal administration advantageously which require puncture of the skin of the subject. For reduces the risk of side effects associated with the systemic example, it is shown in Example 3 that pooled human IgG exposure of IgG. can be efficiently delivered to the brain using nasal drops or a nasal spray. 0119 Advantageously, it was also found that pooled I0123. Another benefit provided by the methods and com human immunoglobulin G was efficiently delivered to the positions provided herein for intranasal administration of brain following intranasal administration in the absence of a pooled human IgG is improved patient compliance. Treat permeability enhancer (e.g., membrane fluidizers, tight junc ment with intravenous IgG (IVIG) requires a lengthy admin tion modulators, and medium chain length fatty acids and istration period under medical Supervision, generally taking salts and esters thereof, as described below). Previous place at hospitals and medical facilities. For example, initial reports have Suggested that in order to achieve efficient administration of IVIG occurs over a 2 to 5 hour period, transport of biotherapeutics (e.g., mimetibodies and Fc once a day for 2 to 7 consecutive days. Follow-up doses, also fusions) through the olfactory epithelium, a permeability typically administered at a hospital over a period of 2 to 5 hours, are required every 1 to 4 weeks depending on the enhancer is required (WO 2009/058957). However, as indication being treated and dosing regimen. Such an admin shown in the examples provided herein, pooled human IgG istration regime is time consuming and inconvenient for is efficiently delivered to the brain when intranasally admin many patients. In comparison, intranasal administration can istered as a liquid or dry powder formulated with only an be administered at home without medical Supervision. Also, amino acid (e.g., glycine). intranasal administration can be performed quickly, over 0120 Advantageously, it is also shown herein that the several minutes depending on the number of dropS/sprays dose of pooled human IgG can be significantly reduced required, rather than several hours as required for intrave when administered intranasally, as compared to intravenous nous administration. Thus, treatment can be prescribed more administration. For example, it is shown in Example 9 that frequently at lower doses to maintain an effective level of administration of a low dose of pooled human IgG (0.002 IgG in the CNS with minimal inconvenience because admin g/kg IgG) intranasally delivered directly to the olfactory istration occurs at home in a shorter period of time. epithelium results in Substantially the same amount of IgG 0.124. Furthermore, IVIG therapy requires catheterization being delivered to the right and left hemispheres of the brain which can cause discomfort and infection at the site of the as for intravenous administration of a ten-fold higher dose of catheter. IVIG solutions are often high in sodium and pooled human IgG (0.02 g/kg IgG, compare corrected AUC glucose to create isotonicity, causing increased risk to the values for right and left hemisphere IgG delivery in Table 71 elderly population, which already have increased rates of and Table 72). A ten-fold reduction in the amount of pooled diabetes and high blood pressure. On the other hand, intra human IgG required for administration is significant because nasal administration is non-invasive, i.e. there is no cath of the limited Supply of pooled human IgG and the high cost eterization and does not carry invasive-procedure related associated therewith. risks such as infection and discomfort at the site of the catheter. Intranasal administration of pooled human IgG 0121 The results described above, which taken together compositions is an improved procedure for elderly persons Suggest that low doses of intranasally administered pooled human IgG is effective for the treatment of Alzheimer's because it does not require IV perfusion and thus does not disease, are Surprising given the difficulty of delivering create a systemic increase in concentrations of Salt or full-length immunoglobulins to the brain via intranasal glucose in the blood. administration. First, although antibody fragments (e.g., 0.125 Thus, as compared to currently utilized modes of Fabs) have previously been administered intranasally, the administering pooled human IgG (e.g., intravenous, Subcu inventors are unaware of any reports demonstrating delivery taneous, and intramuscular) intranasal administration of full-length antibodies to the brain via intranasal admin increases the ease of administration, decreases overall istration. In fact, it has been reported that the delivery of administration time, decreases the number of hospital visits full-length antibodies poses a great difficulty in the field of required, and eliminates the risks associated with catheter medicine (Harmsen M M et al., Appl Microbiol Biotechnol. based administration (e.g., IV administration). Thus, imple 2007, 77(1): 13-22: Athwal G S, Innovations in Pharmaceu mentation of intranasal administration of pooled human IgG tical Technology, July 2009; WO 2006/091332; and WO will result in improved patient compliance. 2009/058957). Consistent with these reports, Applicants found that antibody fragments are delivered much more DEFINITIONS readily to the brain, as compared to full-length immuno 0.126. As used herein, the terms “disorder of the central globulins, after intranasal administration. For example, it is nervous system,” “central nervous system disorder,” “CNS shown in Example 2 that, on average, the concentration of disorder,’ and the like refer to a disorder affecting either the Fabs in brain tissue post-intranasal administration is spinal cord (e.g., a myelopathy) or brain (e.g., an encepha 19-times higher than the concentration of full-length immu lopathy) of a subject, which commonly presents with neu noglobulins post-intranasal administration. Given the sig rological and/or psychiatric symptoms. CNS disorders nificantly lower delivery of full-length immunoglobulins to include many neurodegenerative diseases (e.g., Hunting US 2017/0044244 A1 Feb. 16, 2017

ton's disease, Amyotrophic lateral Sclerosis (ALS), heredi 95%, 96%, 97%, 98%, or 99% IgG (w/w). In certain tary spastic hemiplegia, primary lateral Sclerosis, spinal embodiments, the pooled human IgG composition contains muscular atrophy, Kennedy's disease, Alzheimer's disease, intact IgG immunoglobulins. In other embodiments, the ataxias, Huntington's disease, Lewy body disease, a poly pooled human IgG composition contains IgG fragments, for glutamine repeat disease, and Parkinson's disease) and psy example those prepared by treatment of intact antibodies chiatric disorders (e.g., mood disorders, Schizophrenias, and with trypsin. In certain embodiments, the pooled human IgG autism). Non-limiting examples of ataxia include Friedre compositions used in the treatments disclosed herein contain ich's ataxia and the spinocerebellar ataxias. Specifically for natural or synthetic modifications, e.g., post-translational this application, CNS disorders do not include disorders modifications and/or chemical modifications. resulting from acute viral and bacterial infections. I0131. As used herein, the terms “high titer anti-amyloid 0127. Non-limiting examples of CNS disorders include B pooled immunoglobulin G' and “high titer anti-amyloid? neurodegenerative disorders of the central nervous system, pooled IgG refer to a composition containing polyvalent systemic atrophies primarily affecting the central nervous immunoglobulin G (IgG) purified from the blood/plasma of system, extrapyramidal and movement disorders, demyeli multiple donors, e.g., more than a hundred or more than a nating disorders of the central nervous system, episodic or thousand blood donors, having a relative titer of anti paroxysmal disorders of the central nervous system, para amyloid B immunoglobulin G that is greater than the lytic syndromes of the central nervous system, nerve, nerve expected titer of anti-amyloid B immunoglobulins in a root, or plexus disorders of the central nervous system, pooled IgG composition prepared from the blood/plasma of organic mental disorders, mental or behavioral disorders more than a thousand random individuals. Commercially caused by psychoactive Substance use, Schizophrenic, available intravenous immunoglobulin G (IVIG) prepara Schizotypal, or delusional disorders, mood (affective) disor tions contain IgGs that specifically recognize epitopes of ders, neurotic, stress-related, or somatoform disorders, various conformers of amyloid B, e.g., amyloid B monomers, behavioral syndromes, adult personality or behavior disor amyloid B fibrils, and cross-linked amyloid 3 protein species ders, psychological development disorders, and child onset (CAPS). It has been reported that a commercial preparation behavioral or emotional disorders. (Diagnostic and Statisti of GAMMAGARD LIQUIDR (10% Immune Globulin cal Manual of Mental Disorders, 4th Edition (DSM-IV): The Infusion (Human); Baxter International Inc., Deerfield, Ill.) World Health Organization, The International Classification contains 0.1% anti-amyloid B fibril IgG, 0.04% anti-CAPS of Diseases, 10th revision (ICD-10), Chapter V. Further IgG, and 0.02% anti-amyloid f monomer IgG, having ECso exemplary CNS disorders are provided herein below. affinities of 40 nM, 40 nM, and 350 nM for their target 0128. Neurodegenerative CNS disorders are typically amyloid f conformer, respectively (O'Nuallain B. et al., characterized by progressive dysfunction and/or cell death in Biochemistry, 2008 Nov. 25; 47(47): 12254-6, the content of the central nervous system. The hallmark of many neuro which is hereby incorporated by reference in its entirety for degenerative CNS disorders is the accumulation of mis all purposes). In some embodiments, a high titer anti folded proteins, such as beta-amyloid, tau, alpha-synuclein, amyloid 3 pooled immunoglobulin G composition contains and TDP-43, both intracellularly and extracellularly. Many a high titer of IgG specific for one or more conformer of neurodegenerative diseases are also associated with gross amyloid 3. In other embodiments, a high titer anti-amyloid mitochondrial dysfunction. Common examples of neurode B pooled immunoglobulin G composition contains a high generative CNS disorders include Alzheimer's disease titer of IgG specific for amyloid B monomers, amyloid B (AD), Parkinson's disease (PD), Huntington's disease, and fibrils, and cross-linked amyloid f protein species (CAPS). Amyotrophic lateral sclerosis (ALS). 0.132. Accordingly, in one embodiment, a high titer anti 0129. Psychiatric disorders (also referred to as mental amyloid 3 pooled immunoglobulin G composition contains illnesses) commonly present with cognitive deficits and at least 0.1% anti-amyloid f IgG (e.g., 0.1% IgG with mood dysregulation. Psychiatric disorders are generally specific affinity for any amyloid 3 conformer). In another defined by a combination of how a person feels, acts, thinks embodiment, a high titer anti-amyloid B pooled immuno or perceives. Well established systems for the classification globulin G composition contains at least 0.2% anti-amyloid of psychiatric disorders include the International Statistical B IgG. In yet other embodiments, a high titer anti-amyloid Classification of Diseases and Related Health Problems, B pooled immunoglobulin G composition contains at least 10th Revision (World Health Organization, tenth revision 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, (2010), the content of which is hereby expressly incorpo 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, rated by reference in its entirety for all purposes) and the 0.95%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, Diagnostic and Statistical Manual of Mental Disorders 5.0% or more anti-amyloid f IgG. (DSM-IV: American Psychiatric Association, DS-IV-TR I0133. In one embodiment, a high titer anti-amyloid 3 (2000), the content of which is hereby expressly incorpo pooled immunoglobulin G composition contains at least rated by reference in its entirety for all purposes). Common 0.1% anti-amyloid 3 fibril IgG. In another embodiment, a examples of psychiatric disorders include mood disorders, high titer anti-amyloid 3 pooled immunoglobulin G com Schizophrenia, and autism. position contains at least 0.2% anti-amyloid B fibril IgG. In 0130. As used herein, the terms “pooled human immu yet other embodiments, a high titer anti-amyloid B pooled noglobulin G' and “pooled human IgG” refer to a compo immunoglobulin G composition contains at least 0.15%, sition containing polyvalent immunoglobulin G (IgG) puri 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, fied from the blood/plasma of multiple donors, e.g., more 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, than a hundred or more than a thousand blood donors. 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0% or Typically, the composition will be at least 80% IgG (w/w, more anti-amyloid f fibril IgG. e.g., weight IgG per weight total protein), preferably at least I0134. In one embodiment, a high titer anti-amyloid 3 85%. 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, pooled immunoglobulin G composition contains at least US 2017/0044244 A1 Feb. 16, 2017

0.04% anti-CAPS IgG. In another embodiment, a high titer biological sample from the Subject, or proteins extracted anti-amyloid 3 pooled immunoglobulin G composition con from the biological sample, by mass spectroscopy. In some tains at least 0.08% anti-CAPS IgG. In yet other embodi embodiments, the intactness of the administered immuno ments, a high titer anti-amyloid B pooled immunoglobulin G globulins is determined by separating proteins present in a composition contains at least 0.04%, 0.05%, 0.06%, 0.07%, biological sample from the Subject by molecular weight, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, size, or shape (e.g., by electrophoresis or size exclusion 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, chromatography) and determining the size distribution of 0.8%, 0.85%, 0.9%, 0.95%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, administered immunoglobulins in the sample. 3.5%, 4.0%, 4.5%, 5.0% or more anti-CAPS IgG. 0.138. In one embodiment, the intactness of immuno 0135) In one embodiment, a high titer anti-amyloid 3 globulin (e.g., pooled human IgG) in the brain of a subject pooled immunoglobulin G composition contains at least following intranasal administration is at least 40%. In pre 0.02% anti-amyloid f monomer IgG. In another embodi ferred embodiments, the intactness of immunoglobulin (e.g., ment, a high titer anti-amyloid B pooled immunoglobulin G pooled human IgG) in the brain of a Subject following composition contains at least 0.04% anti-amyloid 3 mono intranasal administration is at least 50%, preferably at least mer IgG. In yet other embodiments, a high titer anti-amyloid 60%. In certain embodiments, the intactness of immuno B pooled immunoglobulin G composition contains at least globulin (e.g., pooled human IgG) in the brain of a subject 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, following intranasal administration is at least 35%, 36%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 0.85%, 0.9%, 0.95%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 57%, 58%, 59%, 60%, 61%, or higher. 4.0%, 4.5%, 5.0% or more anti-amyloid B monomer IgG. 0.139. As used herein, the terms “intranasal administra 0.136 High titer anti-amyloid (B pooled IgG can be tion' and “nasal administration” refer to administration of a prepared according to standard methods for the manufacture therapeutic composition to the nasal cavity of a Subject Such of pooled IgG starting with a standard pool of blood/plasma that a therapeutic agent in the composition is delivered of multiple donors, e.g., more than a hundred or more than directly to one or more epithelium located in the nose. In a thousand blood donors, and Subsequently enriched for certain embodiments, intranasal administration is achieved anti-amyloid B immunoglobulin G. Methods for the enrich using a liquid preparation (e.g., an aqueous preparation), an ment of target-specific immunoglobulin G molecules are aerosolized preparation, or a dry powder preparation, each well known in the art (for example, see U.S. Patent Appli of which can be administered via an externally propelled or cation Publication No. 2004/0101909, the content of which self-propelled (e.g., via inhalation) non-invasive nasal deliv is hereby expressly incorporated by reference herein in its ery device, or via a gel, cream, ointment, lotion, or paste entirety for all purposes). Alternatively, high titer anti directly applied to one or more nasal epithelium (e.g., amyloid B pooled IgG can be prepared according to standard olfactory epithelium or nasal respiratory epithelium). methods for the manufacture of pooled IgG starting with an 0140. As used herein, the term “nasal epithelium” refers enriched pool of blood/plasma from at least fifty, one to the tissues lining the internal structure of the nasal cavity. hundred, two hundred, five hundred, or one thousand donors The term nasal epithelium includes both the nasal respira having a high relative titer of anti-amyloid 3 immunoglobu tory epithelium, located in the lower two-thirds of the nasal lin G. As compared to the manufacture of standard IgG for cavity in humans, and the olfactory epithelium, located in intravenous administration, hyperimmune IgG preparations the upper third of the nasal cavity of humans. are commonly prepared from Smaller donor pools. These 0.141. As used herein, the term “olfactory epithelium’ enriched pools of blood/plasma can be formed, for example, refers to a specialized epithelial tissue inside the nasal cavity by selectively pooling blood/plasma donations or donors involved in smell. In humans, the olfactory epithelium is with a high relative titer of anti-amyloid B immunoglobulin located in the upper third of the nasal cavity. G, e.g., by selection of high titer blood/plasma donations or 0142. As used herein, the term “directed administration” donors. Alternatively, an enriched pool of blood/plasma can refers to a process of preferentially delivering a therapeutic be formed by screening for blood/plasma donations or agent to a first location in a subject as compared a second donors with a low relative titer of anti-amyloid 3 immuno location or systemic distribution of the agent. For example, globulin G and excluding these donations or donors from the in one embodiment, directed administration of a therapeutic starting blood/plasma pool, e.g., Screening for low titer agent results in at least a two-fold increase in the ratio of blood/plasma donations or donors. therapeutic agent delivered to a targeted site to therapeutic 0137 As used herein, the term “intactness” refers to a agent delivered to a non-targeted site, as compared to the percentage of therapeutic agent that has not been at least ratio following systemic or non-directed administration. In partially degraded at a particular point in time following other embodiments, directed administration of a therapeutic administration. In one embodiment, intactness is a measure agent results in at least a 3-fold, 4-fold, 5-fold, 6-fold, of the total administered dose of the therapeutic agent that 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, has not been at least partially degraded at the particular point 30-fold, 35-fold, 40-fold, 45-fold, 50-fold, 60-fold, 70-fold, in time (i.e., systemic intactness). In another embodiment, 80-fold, 90-fold, 100-fold, 200-fold, 300-fold, 400-fold, intactness is a measure of the therapeutic agent present at a 500-fold, 750-fold, 1000-fold, or greater increase in the ratio particular site of the Subject, e.g., brain or bloodstream, of therapeutic agent delivered to a targeted site to therapeutic which has not been at least partially degraded (i.e., local agent delivered to a non-targeted site, as compared to the intactness). In one embodiment, the intactness of adminis ratio following systemic or non-directed administration. In a tered immunoglobulin (e.g., pooled human IgG) is measured particular embodiment, directed administration of an agent by mass spectroscopy. For example, the intactness of the is contrasted to intravenous administration of the agent. For administered immunoglobulins is determined by analyzing a example, in one embodiment, the ratio of therapeutic agent US 2017/0044244 A1 Feb. 16, 2017

present at a targeted site to therapeutic agent present in the lated for intranasal administration which promotes the pas blood stream is increased at least two-fold when the agent is sage of biotherapeutics (e.g., mimetibodies and Fc-fusion Subject to directed administration (e.g., by delivery to the polypeptides) through the nasal epithelium. Non-limiting brain via intranasal administration), as compared to when examples of permeability enhancers include membrane flu the therapeutic agent is administered intravenously. idizers, tight junction modulators, and medium chain length 0143. As used herein, the term “non-invasive nasal deliv fatty acids and salts and esters thereof. Non-limiting ery device' refers an instrument that is capable of delivering examples of medium chain length fatty acids and salts and a therapeutic composition (e.g., pooled human IgG) to the esters thereof included mono-, di-, and triglycerides (such as nasal cavity without piercing the epithelium of the Subject. Sodium caprylate, Sodium caprate, glycerides (CAPMUL. Non-limiting examples of non-invasive nasal delivery GELUCIRE 44/14 PEG32 glyceryl laurate EP); lipids: devices include propellant (e.g., a pressurized inhaler) and pegylated peptides; and liposomes. Surfactants and similarly non-propellant (e.g., a pump-type inhaler) types of aerosol acting compounds can also be used as permeability enhanc or atomizer devices, particle dispersion devices, nebulizers, ers. Non-limiting examples of Surfactants and similarly and pressurized olfactory delivery devices for delivery of acting compounds include polysorbate-80, phosphatidylcho liquid or powder formulations. line, N-methylpiperazine, sodium salicylate, melittin, and 0144. The term “treatment” or “therapy” generally means palmitoyl carnitine chloride (PCC). Generally, the pooled obtaining a desired physiologic effect. The effect may be human immunoglobulin G compositions described herein prophylactic in terms of completely or partially preventing are formulated for intranasal administration in the absence a disease or condition or symptom thereof and/or may be of permeability enhancers. therapeutic in terms of a partial or complete cure for an 0149. As used herein, the term “dry powder composition' injury, disease or condition and/or amelioration of an refers to a lyophilized or spray dried form of a therapeutic adverse effect attributable to the injury, disease or condition pooled human IgG formulation. In one embodiment, a dry and includes arresting the development or causing regres powder composition contains less than 10%, 9%, 8%, 7%, sion of a disease or condition. Treatment can also refer to 6%, 5%, 4%, 3%, 2%, 1%, or less residual water content. any delay in onset, amelioration of symptoms, improvement 0150. A “control is used herein, refers to a reference, in patient Survival, increase in Survival time or rate, usually a known reference, for comparison to an experimen improvement in cognitive function, etc. The effect of treat tal group. One of skill in the art will understand which ment can be compared to an individual or pool of individuals controls are valuable in a given situation and be able to not receiving the treatment. analyze databased on comparisons to control values. Con 0145 As used herein, a “therapeutically effective amount trols are also valuable for determining the significance of or dose” or “sufficient/effective amount or dose.” refers to a data. For example, if values for a given parameter vary dose that produces effects for which it is administered. The widely in controls, variation in test samples will not be exact dose will depend on the purpose of the treatment, and considered as significant. will be ascertainable by one skilled in the art using known 0151. Before the present disclosure is described in greater techniques (see, e.g., Lieberman, Pharmaceutical Dosage detail, it is to be understood that this invention is not limited Forms (vols. 1-3, 1992); Lloyd, The Art, Science and to particular embodiments described, as Such may, of course, Technology of Pharmaceutical Compounding (1999); vary. It is also to be understood that the terminology used Pickar, Dosage Calculations (1999); and Remington: The herein is for the purpose of describing particular embodi Science and Practice of Pharmacy, 20th Edition, 2003, ments only, and is not intended to be limiting, since the Gennaro, Ed., Lippincott, Williams & Wilkins). scope of the present invention will be limited only by the 0146. As used here, the terms “dose” and “dosage' are appended claims. used interchangeably and refer to the amount of active 0152. Where a range of values is provided, it is under ingredient given to an individual at each administration. The stood that each intervening value, to the tenth of the unit of dose will vary depending on a number of factors, including the lower limit unless the context clearly dictates otherwise, frequency of administration; size and tolerance of the indi between the upper and lower limit of that range and any vidual; severity of the condition; risk of side effects; and the other stated or intervening value in that stated range, is route of administration. One of skill in the art will recognize encompassed within the invention. The upper and lower that the dose can be modified depending on the above factors limits of these Smaller ranges may independently be or based on therapeutic progress. The term "dosage form’ included in the Smaller ranges and are also encompassed refers to the particular format of the pharmaceutical, and within the invention, Subject to any specifically excluded depends on the route of administration. For example, a limit in the Stated range. Where the stated range includes one dosage form can be a liquid or dry powder, formulated for or both of the limits, ranges excluding either or both of those intranasal administration. included limits are also included in the invention. 0147 As used herein, a therapeutic composition “con 0153. Unless defined otherwise, all technical and scien sisting essentially of a buffering agent and pooled human tific terms used herein have the same meaning as commonly IgG may also contain residual levels of chemical agents understood by one of ordinary skill in the art to which this used during the manufacturing process, e.g., Surfactants, invention belongs. Although any methods and materials buffers, salts, and stabilizing agents, as well as chemical similar or equivalent to those described herein can also be agents used to pH the final composition, for example, used in the practice or testing of the present invention, counter ions contributed by an acid (e.g., hydrochloric acid representative illustrative methods and materials are now or acetic acid) or base (e.g., Sodium or potassium hydrox described. ide), and/or trace amounts of contaminating proteins. 0154 It is noted that, as used herein and in the appended 0148. As used herein, the term “permeability enhancer' claims, the singular forms “a,” “an,” and “the include plural refers to a component of a therapeutic composition formu referents unless the context clearly dictates otherwise. It is US 2017/0044244 A1 Feb. 16, 2017

further noted that the claims may be drafted to exclude any tive to reduce total amyloid plaque load in a rodent model of optional element. As such, this statement is intended to serve Alzheimer's disease. Moreover, by specifically targeting the as antecedent basis for use of such exclusive terminology as nasal epithelium, as opposed to the respiratory system (lung, “solely,” “only, and the like in connection with the recita pharynx, etc.), systemic exposure of the pooled human tion of claim elements, or use of a “negative' limitation. immunoglobulins is reduced. 0155 As will be apparent to those of skill in the art upon 0159. Many types of intranasal delivery devices can be reading this disclosure, each of the individual embodiments used to practice the methods provided herein. In some described and illustrated herein has discrete components and embodiments, the delivery device is an intranasal device for features which may be readily separated from or combined the administration of liquids. Non-limiting examples of with the features of any of the other several embodiments devices useful for the administration of liquid compositions without departing from the scope or spirit of the present (e.g., liquid pooled IgG compositions) include vapor devices invention. Any recited method can be carried out in the order (e.g., vapor inhalers), drop devices (e.g., catheters, single of events recited or in any other order which is logically dose droppers, multi-dose droppers, and unit-dose pipettes), possible. mechanical spray pump devices (e.g., Squeeze bottles, multi dose metered-dose spray pumps, and single/duo-dose spray Administration pumps), bi-directional spray pumps (e.g., breath-actuated 0156 Intranasal (IN) administration is an advantageous nasal delivery devices), gas-driven spray systems/atomizers mode of delivering a drug to the brain because it is non (e.g., single- or multi-dose HFA or nitrogen propellant invasive and there is a direct connection between the olfac driven metered-dose inhalers, including traditional and cir tory system and the brain. Intranasal administration of IgG cumferential velocity inhalers), and electrically powered (INIG) to treat neurological diseases is particularly advan nebulizers/atomizers (e.g., pulsation membrane nebulizers, tageous because the direct connection between the olfactory vibrating mechanical nebulizers, and hand-held mechanical system and the brain obviates delivery concerns associated nebulizers). In some embodiments, the delivery device is an with the blood-brain barrier (BBB) and minimizes systemic intranasal device for the administration of powders or gels. exposure to the drug, thereby minimizing side effects of the Non-limiting examples of devices useful for the adminis drug. Furthermore, IN delivery allows compositions such as tration of powder compositions (e.g., lyophilized or other powders, granules, solutions, ointments, and creams, wise dried pooled IgG compositions) include mechanical thereby obviating the need for intravenous and intramuscu powder sprayers (e.g., hand-actuated capsule-based powder lar administration. For example, when a drug is administered spray devices and hand-actuated powder spray devices, hand intranasally, it is transported through the nasal mucosa and actuated gel delivery devices), breath-actuated inhalers (e.g., along the olfactory neural pathway. The drug can be admin single- or multi-dose nasal inhalers and capsule-based istered alone or can be combined with a carrier molecule(s) single- or multi-dose nasal inhalers), and insufflators (e.g., to promote transport through the nasal mucosa and along the breath-actuated nasal delivery devices). In some embodi olfactory neural pathway. The drug can also be administered ments, the pooled human immunoglobulins are preferen in combination with an absorption enhancer. Absorption tially administered to the olfactory area, located in the upper enhancers promote the absorption of the drug through the third of the nasal cavity, and particularly to the olfactory nasal mucosa and along the olfactory neural pathway. Fur epithelium. Fibers of the olfactory nerve are unmyelinated thermore, additional molecules can be added to facilitate axons of olfactory receptor cells, which are located in the drug transport across the olfactory neural pathway. superior one-third of the nasal cavity. The olfactory receptor O157. IN administration can also be used to deliver cells are bipolar neurons with swellings covered by hair-like therapeutic drugs to the brain via the trigeminal pathway. cilia that project into the nasal cavity. At the other end, axons Specifically, IN administration can be used to deliver IgG from these cells collect into aggregates and enter the cranial via the trigeminal pathway. The olfactory and trigeminal cavity at the roof of the nose. Surrounded by a thin tube of nerves receive high concentrations of a drug with IN admin pia, the olfactory nerves cross the Subarachnoid space con istration because the absorbent respiratory and olfactory taining CSF and enter the inferior aspects of the olfactory pseudoepithelium are innervated by the trigeminal nerve. bulbs. Once the pooled human immunoglobulin is dispensed These nerves can then transport the drug into the brain and into the nasal cavity, the immunoglobulin can undergo other connected structures. For example, the trigeminal transport through the nasal mucosa and into the olfactory nerve branches directly or indirectly reach the maxillary bulb and interconnected areas of the brain, such as the sinus, brainstem, hindbrain, cribriform plate, forebrain (e.g., hippocampal formation, amygdaloid nuclei, nucleus basalis cortex and diencephalon), orofacial structures (e.g., teeth, of Meynert, locus ceruleus, the brain stem, and the like (e.g., masseter muscle, and the temporomandibular joint), mid Johnson et al., Molecular Pharmaceutics (2010) 7(3):884 brain, cerebellum, cervical spinal cord, thoracic spinal cord, 893). and lumbar spinal cord. Accordingly, INIG can be carried 0160. In certain embodiments, pooled human immuno across the trigeminal pathway to reach and treat neurological globulin is administered to tissue innervated by the trigemi diseases. nal nerve. The trigeminal nerve innervates tissues of a 0158. In certain embodiments, methods are provided for mammals (e.g., human) head including skin of the face and the treatment of CNS disorders by administration of pooled Scalp, oral tissues, and tissues Surrounding the eye. The human immunoglobulins to tissue innervated by the olfac trigeminal nerve has three major branches, the ophthalmic tory and/or trigeminal nerves. Surprisingly, it was found that nerve, the maxillary nerve, and the mandibular nerve. In therapeutically effective amounts of pooled human immu some embodiments, the methods provided herein include noglobulin are delivered to the CNS when administered targeted administration of pooled human immunoglobulin to intranasally. For example, it is shown herein that intranasal one or more of these trigeminal branches, i.e. the trigeminal administration of pooled human immunoglobulins is effec pathway. In some embodiments, the methods provided US 2017/0044244 A1 Feb. 16, 2017

herein include targeted administration of pooled human so that it may be aerosolized for inhalation through the nasal immunoglobulin to the maxillary sinus, thereby reaching the cavity. For example, in one embodiment, the therapeutic brainstem, hindbrain, cribriform plate, forebrain (e.g., cortex agent is placed into a plastic bottle atomizer. In a specific and diencephalon), midbrain, cerebellum, cervical spinal embodiment, the atomizer is advantageously configured to cord, thoracic spinal cord, and lumbar spinal cord through allow a substantial amount of the spray to be directed to the the trigeminal pathway. In certain embodiments, methods upper one-third region or portion of the nasal cavity (e.g., provided herein include targeted administration of pooled the olfactory epithelium). In another embodiment, the liquid human immunoglobulin for treatment of a disorder of the preparation is aerosolized and applied via an inhaler, such as CNS (e.g., Alzheimer's disease). a metered-dose inhaler (for example, see, U.S. Pat. No. 0161 In some embodiments, the pooled human immu 6,715.485). In a specific embodiment, the inhaler is advan noglobulin is administered to nasal tissues innervated by the trigeminal nerve, for example, to nasal tissues including the tageously configured to allow a Substantial amount of the sinuses, the inferior two-thirds of the nasal cavity and the aerosol to be directed to the upper one-third region or nasal septum. In certain embodiments, the pooled human portion of the nasal cavity (e.g., the olfactory epithelium). In immunoglobulin is targeted to the inferior two-thirds of the certain embodiments, a Substantial amount of the pooled nasal cavity and/or the nasal septum. human immunoglobulin refers to at least 25%, 30%, 35%, 0162. In some embodiments, the pooled human immu 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, noglobulin is administered to one or both maxillary sinus of 90%. 95%, or 100% of the composition, which is adminis the individual. Methods and devices for administration to tered to the upper one-third region of the nasal cavity (e.g., the maxillary sinus are known in the art, for example, see administered to the upper one-third of the nasal epithelium). United States Patent Application Publication Number 2011/ 0167. In one embodiment, the pooled human IgG com 0151393, the contents of which are hereby incorporated by positions provided herein for the treatment of a CNS disor reference in their entirety for all purposes. der (e.g., Alzheimer's disease) are intranasally administered 0163 The maxillary sinus is in fluid communication with as a dry powder. Dry powder nasal delivery devices are well the patient's nasal cavity and comprises right and left known in the art, for example, see PCT publication No. WO maxillary sinuses. Each maxillary sinus communicates with 1996/222802. In one embodiment, following intranasal the corresponding nasal passage via the orifice of the max administration, pooled human IgG is absorbed across the illary sinus. The maximum Volume of the maxillary sinus in olfactory epithelium, which is found in the upper third of the adults is approximately 4 to 15 ml, though individual sinuses nasal cavity. In another embodiment, following intranasal may comprise Volumes outside of this range. administration, pooled human IgG is absorbed across the 0164. The pathway from the nasal passages to the corre nasal respiratory epithelium, which is innervated with sponding orifice of maxillary sinus, and ultimately to the trigeminal nerves, in the lower two-thirds of the nasal cavity. corresponding maxillary sinus, allows for a device to be The trigeminal nerves also innervate the conjunctive, oral inserted into the nasal passage to the orifice of the maxillary mucosa, and certain areas of the dermis of the face and head, sinus, whereupon at least one effective amount or dose of and absorption after intranasal administration of the IgG pooled human immunoglobulins may be administered and from these regions may also occur. In other embodiments, delivered into the maxillary sinus. The pathway to the following intranasal administration, pooled human IgG is maxillary sinus is tortuous and requires: traversing the absorbed across the maxillary sinus epithelium. In yet other nostril, moving through the region between the lower and embodiments, pooled human IgG may be absorbed across middle concha, navigating over and into the semilunar more than one of these nasal epitheliums and Subsequently hiatus, traveling Superiorly into the maxillary sinus opening, delivered to the brain of the subject. resisting the ciliated action of the ostium/tube passing into the maxillary sinus and ultimately moving into the sinus 0168 Although administration is referred to herein as a itself. single event that may occur according to Some regular or 0.165 Since the trigeminal nerve passes through the max irregular frequency of the course of a treatment, a single illary sinus, the pooled human immunoglobulins in the administration even may include multiple administrations. maxillary sinus after delivery therein will be moved along In this regard, a single dosage of pooled human IgG may be the trigeminal nerve to structures innervated by the trigemi partitioned into two or more physical compositions for nal nerve. In this fashion, pooled human IgG administered to administration. For example, a 200 mg dose of pooled one or both of the maxillary sinus is delivered to the brain human IgG in a liquid composition formulated at 200 g/L via the trigeminal nerve. IgG may be administered to a 50 kg Subject (4 mg/kg IgG) 0166 In one embodiment, the pooled human IgG com in four drops having a Volume of 250 uL each. Likewise, a positions provided herein for the treatment of a CNS disor dry powder composition containing a single dosage of der (e.g., Alzheimer's disease) are intranasally administered pooled human IgG may be administered, for example, in two as a liquid preparation, e.g., an aqueous based preparation. or more distinct puffs. In some embodiments, pooled human For example, in one embodiment, nasal drops are instilled in IgG is administered in one or more puffs or sprays into each the nasal cavity by tilting the head back sufficiently and nare of the individual (e.g., one or more puff into the right apply the drops into the nares. In another embodiment, the nare and one or more puffs into the left nare). drops are Snorted up the nose. In another embodiment, nasal (0169. In certain embodiments, the methods described drops are applied with an applicator or tube onto the upper herein for treating a CNS disorder include intranasal admin third of the nasal mucosa. In another embodiment, nasal istration of pooled human IgG via a non-invasive intranasal drops are applied with an applicator or tube into one or both delivery device. In one embodiment, the non-invasive intra of maxillary sinus of the subject. In another embodiment, the nasal delivery device is a non-propellant type aerosol or liquid preparation may be placed into an appropriate device atomizer device, a propellant type aerosol or atomizer US 2017/0044244 A1 Feb. 16, 2017

device, a non-propellant pump-type device, a particle dis 0.175. In certain embodiments, the pooled human immu persion device, a nebulizer device, or a pressurized olfactory noglobulin compositions are administered by a pressurized delivery device. nasal delivery (PND) device. In one embodiment, the PND 0170. In one embodiment the non-invasive intranasal device can be used to deliver a liquid IgG composition to the delivery device delivers a liquid drop of a pooled human IgG nasal cavity. In one embodiment, the PND device can be composition to the nasal cavity of a subject. In a particular used to deliver a powder IgG composition to the nasal embodiment, the non-invasive intranasal delivery device cavity. In one embodiment, the PND device administers an delivers a liquid drop of pooled human IgG directly to a IgG composition into one nostril. In one embodiment, the nasal epithelium of the Subject. In a more specific embodi Impel device administers an IgG composition into both ment, the non-invasive intranasal delivery device delivers a nostrils. liquid drop of pooled human IgG directly to the olfactory 0176). In some embodiments, the PND device is config epithelium of the subject. In one embodiment, the liquid ured to deliver the liquid or powder IgG compositions to a drop is administered by tilting the head of the subject back particular epithelium, location, and/or structure of the nasal and administering the drop into a nare of the Subject. In cavity. For example, in one embodiment, the PND device is another embodiment, the liquid drop is administered by configured to deliver the IgG composition to the upper nasal inserting the tip of a non-invasive intranasal delivery device cavity. In one embodiment, the PND device is configured to into a nare of the Subject and squirting or spraying the drop deliver the IgG composition to the olfactory epithelium of into the nasal cavity of the subject. the nasal cavity. In one embodiment, the PND device is 0171 In another embodiment, the non-invasive intrana configured to deliver the IgG composition to the lower two sal delivery device delivers a liquid or a powder aerosol of thirds of the nasal epithelium. In one embodiment, the PND a pooled human IgG composition to the nasal cavity of a device is configured to deliver the IgG composition to a Subject. In a particular embodiment, the non-invasive intra nasal epithelium associated with trigeminal nerve endings. nasal delivery device delivers a liquid or a powder aerosol In one embodiment, the PND device is configured to deliver of pooled human IgG directly to a nasal epithelium of the the IgG composition to the nasal maxillary sinus epithelium. Subject. In a more specific embodiment, the non-invasive 0177 Methods for configuring pressurized delivery intranasal delivery device delivers a liquid or a powder devices to achieve a particular delivery profile are known in aerosol of pooled human IgG directly to the olfactory the field. For example, in one embodiment, a pressurized epithelium of the subject. nasal delivery device is configured to produce a stream, 0172. In another embodiment, the non-invasive intrana spray, puff, etc., have a particular characteristic. For sal delivery device delivers a dry powder composition of example, in one embodiment, to achieve administration to pooled human IgG composition to the nasal cavity of a the upper third of the nasal epithelium, the device is con Subject. In a particular embodiment, the non-invasive intra figured to produce a strong, focused stream, spray, puff, etc. nasal delivery device delivers a dry powder composition of In one embodiment, the strong focused spray is created by pooled human IgG directly to a nasal epithelium of the imparting circumferential and/or axial velocity onto the Subject. In a more specific embodiment, the non-invasive stream of the therapeutic composition (e.g., pooled human intranasal delivery device delivers a dry powder composi IgG) being administered into the nose. In another embodi tion of pooled human IgG directly to the olfactory epithe ment, to achieve administration to a greater portion of the lium of the subject. nasal epithelium (e.g., the entire or the lower two thirds of 0173. In another embodiment, the non-invasive intrana the nasal epithelium), the device is configured to produce a sal delivery device delivers a sustained release or controlled diffuse and/or weaker stream, spray, puff, etc. In some release composition of pooled human IgG composition to embodiments, the tip of the delivery device is configured to the nasal cavity of a Subject. In a specific embodiment the physically direct the stream, spray, puff, etc., to the desired Sustained release composition comprises a dry powder com intranasal location when inserted into the subject’s nare. For position of pooled human IgG. In some embodiments, the example, a kink or bend may be introduced into the tip of the Sustained release composition is a gel, paste, hydrogel, delivery device to “point the stream, spray, puff, etc., at a cream, lotion, film, or similar form that coats at least a targeted epithelium. In some embodiments, the delivery portion of the nasal epithelium (e.g., all or a portion of the pattern of the device is adjustable, such that the device can olfactory epithelium, all or a portion of a nasal epithelium be differentially configured to target the therapeutic agent associated with trigeminal nerve endings, all or a portion of (e.g., pooled human IgG) to a particular epithelium, struc the upper third of the nasal epithelium, all or a portion of the ture, or location within the nose. In certain embodiments, the lower third of the nasal epithelium, or all or a portion of the pooled human immunoglobulin compositions are adminis nasal maxillary epithelium. tered by a breath-powered technology device. In certain 0174. In one embodiment, the intranasal device is a embodiments, the breath-powered technology provides single-use, disposable device. In another embodiment, the positive pressure during administration. In certain embodi intranasal device is a multi- or repeat-use device. In certain ments, the positive pressure expands narrow nasal passages. embodiments, the single-use or multi-use device is pre In certain embodiments, the expansion of the nasal passages metered. In a specific embodiment, the single-use or multi allows reliable delivery of liquid or powder pooled human use device is pre-filled. In certain embodiments, the multi immunoglobulin compositions described herein to the CNS. or repeat-use device is refillable. In certain embodiments, In some embodiments, exhalation into the device propels the the device is refilled by inserting a pooled human IgG therapeutic (e.g., pooled human IgG) into the nose, while at composition into a chamber of the device. In other embodi the same time closing the Soft-palette, thereby reducing ments, a chamber of the multi- or repeat-use device designed deposition of the therapeutic into the throat and/or lungs. In to hold the pooled human IgG composition is replaced with one embodiment, the breath-powered technology device a new, pre-filled chamber. administers an IgG composition described herein into one US 2017/0044244 A1 Feb. 16, 2017

nostril. In one embodiment, the breath-powered technology ments the initial dose is one, two, three, four, five, six, seven, device administers an IgG composition described herein into eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, two nostrils. sixteen, seventeen, eighteen, nineteen, or twenty or more 0178. Non-limiting examples of commercial intranasal doses. delivery devices include the EQUADEL(R) nasal spray pump 0182. In another embodiment, a gel, cream, ointment, (Aptar Pharma), the Solovent dry powder device (BD Tech lotion, or paste containing pooled human IgG is applied onto nologies), the Unidose nasal drug delivery device (Consort the nasal epithelium, for example, by use of an application Medical PLC), the NasoNeb(R) Nasal Nebulizer (MedInvent, Stick or Swab. In a particular embodiment, a gel, cream, LLC), the Veriloser R nasal delivery device (Mystic Phar ointment, lotion, or paste containing pooled human IgG is maceuticals), the VRX2TM nasal delivery device (Mystic applied directly onto a nasal epithelium of the Subject. In a Pharmaceuticals), the DirectHalerTM Nasal device (Direct more specific embodiment, a gel, cream, ointment, lotion, or Haler A/S), the TriViarTM single-use unit-dose dry powder paste containing pooled human IgG is applied directly onto inhaler (Trimel Pharmaceuticals), the SinuStarTM Aerosol the olfactory epithelium of the subject. Delivery System (Pari USA), the Aero Pump (Aero Pump 0183. In certain embodiments, a substantial fraction of GmbH), the Fit-LizerTM nasal delivery device (Shin Nippon the therapeutic agent present in the composition is delivered Biomedical Laboratories), the LMA MAD NasalTM device directly to one or more nasal epithelium. In certain embodi (LMA North America, Inc.), the Compleo intranasal bioad ments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, hesive gel delivery system (Trimel Pharmaceuticals), 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the Impel's Pressurized Olfactory Delivery (POD) device (Im therapeutic agent present in the composition is delivered pel Neuropharma), the ViaNaseTM electronic atomizer directly to a nasal epithelium. In a specific embodiment, a (Kurve Technology, Inc.), the OptiNose powder delivery Substantial fraction of the therapeutic agent present in the device (OptiNose US Inc.), and the Optinose liquid delivery composition is delivered directly to the olfactory epithelium. device (OptiNose US Inc.) In a more specific embodiment, at least 25%, 30%, 35%, 0179. In one embodiment, an intranasal device described 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, herein can deliver 10%-20% of the metered IgG dose to the 90%. 95%, or 100% of the therapeutic agent present in the olfactory region. In one embodiment, an intranasal device composition is delivered directly to the olfactory epithelium. described herein can deliver 20%-30% of the metered IgG In another specific embodiment, a substantial fraction of the dose to the olfactory region. In one embodiment, an intra therapeutic agent present in the composition is delivered nasal device described herein can deliver 5%-20% of the directly to nasal epithelium innervated with trigeminal metered IgG dose to the olfactory region. In one embodi nerves. In a more specific embodiment, at least 25%, 30%, ment, an intranasal device described herein can deliver 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 30%-40% of the metered IgG dose to the olfactory region. 85%, 90%, 95%, or 100% of the therapeutic agent present in In one embodiment, an intranasal device described herein the composition is delivered directly to nasal epithelium can deliver 40%-50% of the metered IgG dose to the innervated with trigeminal nerves. olfactory region. In one embodiment, an intranasal device 0184. In some embodiments, pooled human IgG can be described herein can deliver 60%–70% of the metered IgG administered to a subject as a combination therapy with dose to the olfactory region. In one embodiment, an intra another treatment, e.g., another treatment for a disorder of nasal device described herein can deliver 60%-80% of the the central nervous system (e.g., Alzheimer's disease, age metered IgG dose to the olfactory region. In one embodi related dementia, Parkinson's disease, or multiple Sclerosis). ment, an intranasal device described herein can deliver For example, the combination therapy can include admin 70%–80% of the metered IgG dose to the olfactory region. istering to the Subject (e.g., a human patient) one or more In one embodiment, an intranasal device described herein additional agents that provide a therapeutic benefit to the can deliver 80%-90% of the metered IgG dose to the Subject who has, or is at risk of developing, a disorder of the olfactory region. In one embodiment, an intranasal device central nervous system, e.g., Alzheimer's disease. In some described herein can deliver 60%-80% of the metered IgG embodiments, the pooled human IgG and the one or more dose to the olfactory region. additional agents are administered at the same time. In other 0180. In certain embodiments, the pooled human immu embodiments, the pooled human IgG is administered first in noglobulin compositions are administered by an intranasal time and the one or more additional agents are administered device described above in one or more doses. In one second in time. In some embodiments, the one or more embodiment the more than one dose is administer by the additional agents are administered first in time and the intranasal device in alternating nostrils. In one embodiment, pooled human IgG is administered second in time. the more than one does is administered by the intranasal 0185. The pooled human IgG can replace or augment a device at different time points throughout the day. In certain previously or currently administered therapy. For example, embodiments the more than one dose is two, three, four, five, upon treating with pooled human IgG, administration of the six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, one or more additional agents can cease or diminish, e.g., be fifteen, sixteen, seventeen, eighteen, nineteen, or twenty or administered at lower levels. In other embodiments, admin more doses. In certain embodiments the more than one dose istration of the previous therapy is maintained. In some is administered by the intranasal device one, two, three, four, embodiments, a previous therapy will be maintained until five, six, seven, eight, nine, or ten or more time points the level of polyclonal IgG reaches a level sufficient to throughout the day. provide a therapeutic effect. The two therapies can be 0181. In certain embodiments, the pooled human immu administered in combination. noglobulin compositions are administered by an intranasal 0186. In one embodiment, a human receiving a first device described above in an initial dose or set of doses therapy for a disorder of the central nervous system, e.g., followed by repeat maintenance doses. In certain embodi Alzheimer's disease, who is then treated with pooled human US 2017/0044244 A1 Feb. 16, 2017

IgG, continues to receive the first therapy at the same or a 0.4 g/kg to 0.5 g/kg body weight IVIG. Due to the limited reduced amount. In another embodiment, treatment with the Supply of pooled human IgG, and high cost associated first therapy overlaps for a time with treatment with pooled therewith, large-scale implementation of these treatments human IgG, but treatment with the first therapy is subse may prove problematic if they are approved by major quently halted. regulatory bodies. 0187. In a particular embodiment, pooled human IgG 0193 Typical intravenous dosing of IgG in human may be administered in combination with a treatment for an Alzheimer's trials ranges from 200 mg/kg to 400 mg/kg age-related dementia, e.g., Alzheimer's disease. In certain every two weeks. Advantageously, the inventors have found embodiments, the treatment for an age-related dementia that levels of pooled human IgG seen in the brain after co-administered with pooled human IgG is administration of intravenous administration can also be achieved by intrana a cholinesterase inhibitor (e.g., ARICEPT (donepezil), sal administration. For example, it is shown in Example 3 EXELON (rivastigmine), RAZADYNE (galantamine), or that administration of pooled human IgG (0.02 g/kg IgG) COGNEX (tacrine), or an inhibitor of the NMDA-type intranasally as drops (IN1) or a liquid spray delivered glutamate receptor (e.g., memantine). directly to the olfactory epithelium (IN3) results in substan 0188 In further embodiments the second therapy is tially the same amount of IgG being delivered to the right levodopa (L-DOPA). The second therapy can also be a and left hemispheres of the brain as for intravenous admin dopamine agonist. Non-limiting examples of dopamine ago istration of pooled human IgG (0.02 g/kg IgG; compare nists include bromocriptine, pergolide, pramipexole, ropini corrected AUC values for right and left hemisphere IgG role, piribedil, cabergoline, apomorphine and lisuride. The delivery in Table 69, Table 71, and Table 72). Significantly, second therapy can be a MAO-B inhibitor. Non-limiting intranasal administration of IgG liquid drops at concentra examples of MAO-B inhibitors are selegiline and rasgiline. tions ten-fold lower (0.002 g/kg IgG) also resulted in the Addition second therapies can include amantaine, anticho delivery of intact IgG to the cerebral cortex (see, Table 70). linergic compositions, clozapine, modafinil, and non-steroi Any reduction in the amount of pooled human IgG required dal anti-inflammatory drugs. for administration is significant because of the limited 0189 In further embodiments the second therapy is Supply of pooled human IgG and the high cost associated CAMPATH (alemtuzumab), ZENAPX (daclizumab), ritux therewith. imab, dirucotide, BHT-3009, cladribine, dimethyl fumarate, 0194 Accordingly, in certain embodiments, the methods estriol, laquinimod, pegylated interferon-B-1a, minocycline, for treating a CNS disorder provided herein include intra statins, temsirolimus, teriflunomide, and low dose naltex nasally administering from about 0.05 mg of pooled human OC. IgG per kg body weight (mg/kg IgG) to about 500 mg/kg 0190. In certain embodiments the second therapy is psy IgG in a single dosage. chotherapy. Non-limiting examples of psychotherapy are 0.195. In certain embodiments, the methods for treating a psychosocial intervention, behavioral intervention, reminis CNS disorder provided herein include intranasally admin cence therapy, validation therapy, Supportive psychotherapy, istering a low dose of pooled human IgG. In one embodi sensory integration, simulated presence therapy, cognitive ment, a low dose of pooled human IgG is from about 0.05 retraining, and stimulation-oriented therapies Such as art, mg/kg IgG to about 10 mg/kg IgG. In specific embodiments, music, pet, exercise, and recreational activities. a low dose of pooled human IgG is about 0.05 mg/kg, 0.06 0191) Furthermore, two or more second therapies can be mg/kg, 0.07 mg/kg, 0.08 mg/kg, 0.09 mg/kg, 0.10 mg/kg, combined with therapeutic intranasal IgG. For example, 0.15 mg/kg, 0.20 mg/kg, 0.25 mg/kg, 0.30 mg/kg, 0.35 therapeutic intranasal IgG can be combined with memantine mg/kg, 0.40 mg/kg, 0.45 mg/kg, 0.50 mg/kg, 0.55 mg/kg, and donepezil. 0.60 mg/kg, 0.65 mg/kg, 0.70 mg/kg, 0.75 mg/kg, 0.80 mg/kg, 0.85 mg/kg, 0.90 mg/kg, 0.95 mg/kg, 1.0 mg/kg, 1.5 Dosing mg/kg, 2.0 mg/kg, 2.5 mg/kg, 3.0 mg/kg, 3.5 mg/kg, 4.0 0.192 The use of intravenous immunoglobulin G (IVIG) mg/kg, 4.5 mg/kg, 5.0 mg/kg, 6.0 mg/kg, 7.0 mg/kg, 8.0 for the treatment of disorders of the central nervous system mg/kg, 9.0 mg/kg, or 10.0 mg/kg IgG. In yet other embodi (CNS) is currently under investigation (Awad et al. 2011 ments, a low dose of pooled human IgG is from 0.1 mg/kg (Current Neuropharmacology, 9:417428); Pohl et al. 2012 to 5 mg/kg, 0.5 mg/kg to 5 mg/kg, 1 mg/kg to 5 mg/kg, 2 (Current Treatment Options in Neurology, 14:264-275); mg/kg to 5 mg/kg, 0.5 mg/kg to 10 mg/kg, 1 mg/kg to 10 Krause et al. 2012 (European J. of Paediatric Neurology, mg/kg, 2 mg/kg to 10 mg/kg, 1 mg/kg to 8 mg/kg, 2 mg/kg 16:206-208); Elovaara et al. 2011 (Clinical Neuropharma to 8 mg/kg, 3 mg/kg to 8 mg/kg, 4 mg/kg to 8 mg/kg, 5 cology, 34(2):84-89); Perlmutter, et al. 1999 (The Lancet, mg/kg to 8 mg/kg, 1 mg/kg to 6 mg/kg, 2 mg/kg to 6 mg/kg, 354:1153-1158); Snider et al. 2003 (J. of Child and Adoles 3 mg/kg to 6 mg/kg, 4 mg/kg to 6 mg/kg, 5 mg/kg to 6 cent Psychopharmacology, 13(supp 1): S81-S88). In these mg/kg, 1 mg/kg to 4 mg/kg, 2 mg/kg to 4 mg/kg, or 3 mg/kg trials, Subjects are administered between 0.4 g/kg body to 4 mg/kg IgG. weight and 2.0 g/kg body weight IVIG per dosage. Specifi 0196. In certain embodiments, the methods for treating a cally, the treatment regimes of CNS disorders with IVIG CNS disorder provided herein include intranasally admin range from 0.4 g/kg body weight IVIG administered once istering a medium dose of pooled human IgG. In one daily for 5 consecutive days to 2.0 g/kg body weight IVIG embodiment, a medium dose of pooled human IgG is from administered once daily for 2 consecutive days. There are about 10 mg/kg IgG to about 100 mg/kg IgG. In specific several variations of these IVIG treatment regimes. For embodiments, a medium dose of pooled human IgG is about example, IVIG treatment regimes may be 1.0 g/kg body 10 mg/kg, 11 mg/kg, 12 mg/kg, 13 mg/kg, 14 mg/kg, 15 weight IVIG administered twice a day (total 2.0 g/kg body mg/kg, 16 mg/kg, 17 mg/kg, 18 mg/kg, 19 mg/kg, 20 mg/kg, weight IVIG per day). The initial 2 to 5 day IVIG dosages 21 mg/kg, 22 mg/kg, 23 mg/kg, 24 mg/kg, 25 mg/kg, 26 can also be followed with maintenance doses ranging from mg/kg, 27 mg/kg, 28 mg/kg, 29 mg/kg, 30 mg/kg, 31 mg/kg, US 2017/0044244 A1 Feb. 16, 2017

32 mg/kg, 33 mg/kg, 34 mg/kg, 35 mg/kg, 36 mg/kg, 37 may require a high dose and/or high dosage frequency. In yet mg/kg, 38 mg/kg, 39 mg/kg, 40 mg/kg, 41 mg/kg, 42 mg/kg, another embodiment, a subject having a high likelihood of 43 mg/kg, 44 mg/kg, 45 mg/kg, 46 mg/kg, 47 mg/kg, 48 developing a CNS disorder may also be prescribed a low mg/kg, 49 mg/kg, 50 mg/kg, 55, 60, 65, 70, 75, 80, 85,90, dose and/or low dosing frequency as a prophylactic treat 95, or 100 mg/kg IgG. In yet other embodiments, a medium ment or to delay onset of symptoms associated with a CNS dose of pooled human IgG is from 10 mg/kg to 100 mg/kg, disorder. For example, a subject with a familial history of an 25 mg/kg to 100 mg/kg, 50 mg/kg to 100 mg/kg, 75 mg/kg age-related dementia (e.g., Alzheimer's disease) may be to 100 mg/kg, 10 mg/kg to 75 mg/kg, 25 mg/kg to 75 mg/kg, intranasally administered pooled human IgG at a low dosage 50 mg/kg to 75 mg/kg, 10 mg/kg to 50 mg/kg, 25 mg/kg to and/or low frequency to delay the onset of symptoms 50 mg/kg, or 10 mg/kg to 25 mg/kg IgG. associated with the age-related dementia. A skilled physician will readily be able to determine an appropriate dosage and 0197) In some embodiments, the methods for treating a dosing frequency for a Subject diagnosed with or having a CNS disorder provided herein include intranasally admin high likelihood of developing a CNS disorder. istering a high dose of pooled human IgG. In one embodi 0201 In one embodiment, where the progression of a ment, a high dose of pooled human IgG is from about 100 particular CNS disorder in a subject requires frequent dos mg/kg IgG to about 400 mg/kg IgG. In specific embodi ing, the methods provided herein for treating a disorder of ments, a high dose of pooled human IgG is about 100 mg/kg, the central nervous system include administering a compo 110, 120 mg/kg, 130 mg/kg, 140 mg/kg, 150 mg/kg, 175 sition comprising pooled human immunoglobulin G (IgG) to mg/kg, 200 mg/kg, 225 mg/kg, 250 mg/kg, 275 mg/kg, 300 the subject at least once a week. In other embodiments, the mg/kg, 325 mg/kg, 350 mg/kg, 375 mg/kg, 400 mg/kg, or method includes administering a composition comprising higher. In yet other embodiments, a high dose of pooled pooled human immunoglobulin G (IgG) to the Subject at human IgG is from 100 mg/kg to 400 mg/kg, 150 mg/kg to least two, three, four, five, or six times a week. In yet another 400 mg/kg, 200 mg/kg to 400 mg/kg, 250 mg/kg to 400 embodiment, the method includes administering a compo mg/kg, 300 mg/kg to 400 mg/kg, 350 mg/kg to 400 mg/kg, sition comprising pooled human immunoglobulin G (IgG) to 100 mg/kg to 300 mg/kg, 150 mg/kg to 300 mg/kg, 200 the subject at least once daily. In other embodiments, the mg/kg to 300 mg/kg, 250 mg/kg to 300 mg/kg, 100 mg/kg method includes administering a composition comprising to 200 mg/kg, 150 mg/kg to 200 mg/kg, or 100 mg/kg to 150 pooled human immunoglobulin G (IgG) to the Subject at mg/kg IgG. least two, three, four, five, or more times daily. In a specific 0198 In some embodiments, pooled human IgG is embodiment, the CNS disorder is an age-related dementia, administered at a set dosage, regardless of the weight of the Parkinson's disease, or multiple sclerosis. In a more specific subject. Without being bound by theory, unlike intravenous embodiment, the CNS disorder is Alzheimer's disease. administration, the final concentration of IgG in the brain 0202 In another embodiment, where the progression of a should be independent of total body weight when adminis particular CNS disorder in a subject requires less frequent tered intranasally since the therapeutic will travel directly dosing, the methods provided herein for treating a disorder from the nose to the brain. Accordingly, a standard dose of of the central nervous system include administering a com intranasal pooled human IgG, which is independent of body position comprising pooled human immunoglobulin G (IgG) weight, may simplify the process of dosing individual Sub to the Subject at least once a month. In other embodiments, jects. the method includes administering a composition compris 0199 Accordingly, in one embodiment, the methods ing pooled human immunoglobulin G (IgG) to the Subject at described herein include intranasal administration of a fixed least two, three, four, five, six, or more times a month. In yet dose of pooled human IgG of from about 50 mg to about 10 another embodiment, the method includes administering a g. In some embodiments, the fixed dose of IgG is about 50 composition comprising pooled human immunoglobulin G mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 (IgG) to the subject at least once daily. In other embodi mg, 250 mg, 275 mg, 300 mg. 350 mg. 400 mg. 450 mg, 500 ments, the method includes administering a composition mg, 550 mg. 600 mg, 650 mg, 700 mg, 750 mg. 800 mg. 850 comprising pooled human immunoglobulin G (IgG) to the mg, 900 mg, 950 mg, 1.0 g, 1.25 g, 1.5 g, 1.75 g, 2.0 g, 2.5 Subject at least two, three, four, five, or more times daily. In g, 3.0 g, 3.5g, 4.0 g, 4.5 g. 5.0 g, 5.5 g. 6.0 g. 6.5 g, 7.0 g, a specific embodiment, the CNS disorder is an age-related 7.5g, 8.0 g, 8.5g, 9.0 g, 9.5g, 10.0g, or more IgG. In other dementia, Parkinson's disease, or multiple Sclerosis. In a embodiments, the methods described herein include intra more specific embodiment, the CNS disorder is Alzheimer's nasal administration of from 50 mg to 5 g, 100 mg to 5 g, disease. 250 mg to 5 g, 500 mg to 5 g, 750 mg to 5g, 1 g to 5g, 2.5 0203. In certain embodiments, the composition can be g to 5 g, 50 mg to 2.5 g., 100 mg to 2.5g, 250 mg to 2.5 g. administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 500 mg to 2.5g, 750 mg to 2.5 g, 1 g to 2.5 g. 50 mg to 1 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or g, 100 mg to 1 g, 250 mg to 1 g, 500 mg to 1 g, 750 mg to 31 times a month. The composition can be administered 1 g, 50 mg to 500 mg, 100 mg to 500 mg, 250 mg to 500 mg. between equally spaced days of the month, for example, on 50 mg to 250 mg, 100 mg to 250 mg. or 50 mg to 100 mg the 1 and the 15" of each month. Alternatively, the com pooled human IgG. position can be administered in block dosing at the begin 0200. Depending upon the CNS disorder being treated ning, end, or middle of the month. For example, the com and the progression of the disorder in the Subject, the pooled position can be administered only on the 1, 1'-2', 1'-3', human IgG compositions described herein are intranasally 1-4". 1'-5", 1'-6", or 1-7 days of the month. Similar administered to a Subject anywhere from several times daily dosing schemes can be administered toward the middle or to monthly. For example, a subject diagnosed with a CNS end of the month. disorder in an early stage of progression may require only a 0204. In certain embodiments the dosing can change low dosage and/or low dosage frequency, while a subject between dosing days. For example, on the first day of dosing diagnosed with a CNS disorder in a late stage of progression a subject can receive 10 mg/kg IgG and on the second day US 2017/0044244 A1 Feb. 16, 2017 19 of dosing the Subject can receive 20 mg/kg IgG. Similarly, Sally administering from 0.05 mg/kg to 50 mg/kg pooled a Subject who is administered two or more doses per day of human immunoglobulin to a subject in need thereof daily. In intranasal IgG can receive two different doses. For example, other embodiments, the methods provided herein for the the first dose of the day can be 10 mg/kg IgG and the second treatment of a CNS disorder include intranasally adminis dose of the day can be 5 mg/kg IgG. tering pooled human IgG in a dosage/frequency combination 0205. In certain embodiments, the methods provided selected from variations 1 to 816 found in Table 1 and Table herein for the treatment of a CNS disorder include intrana 2. TABLE 1. Exemplary combinations of dosage and frequency for methods of treating a CNS disorder by intranasal administration of pooled human IgG. Two Three Four One Two Three Every One Time Times Times Times Time Times Times Other Monthly Monthly Monthly Monthly Weekly Weekly Weekly Day

O.OS Kg War. 1 War. 52 War. 103 War. 154 War. 205 War. 256 War. 307 War. 358 O.1 Kg War. 2 War. S3 War. 104 War. 155 War. 206 War. 257 War. 308 War. 359 O.25 Kg War. War. 54 War. 105 War. 156 War. 207 War. 258 War. 309 War. 360 Kg War. War. SS War. 106 War. 157 War. 208 War. 259 War. 310 War. 361 0.75 Kg War. War. 56 War. 107 War. 158 War. 209 War. 260 War. 311 War. 362 1.O Kg War. War. S7 War. 108 War. 159 War. 210 War. 261 War. 312 War. 363 1.5 Kg War. War. 58 War. 109 War. 16O War. 211 War. 262 War. 313 War. 364 2.0 Kg War. War. S9 War. 110 War. 161 War. 212 War. 263 War. 314 War. 365 2.5 Kg War. War. 60 War. 111 War. 162 War. 213 War. 264 War. 315 War. 366 3.0 Kg War. War. 61 War. 112 War. 163 War. 214 War. 265 Var. 316 War. 367 3.5 Kg War. War. 62 War. 113 War. 164 War. 215 War. 266 War. 317 War. 368 4.0 Kg War. War. 63 War. 114 War. 165 War. 216 War. 267 War. 318 War. 369 4.5 Kg War. War. 64 War. 115 War. 166 War. 217 War. 268 War. 319 War. 370 Kg War. War. 65 War. 116 War. 167 War. 218 War. 269 War. 32O War. 371 6.O Kg War. War. 66 War. 117 War. 168 War. 219 War. 270 War. 321 War. 372 7.0 Kg War. War. 67 War. 118 War. 169 War. 22O War. 271 War. 322 War. 373 8.0 Kg War. War. 68 War. 119 War. 17O War. 221 War. 272 War. 323 War. 374 9.0 Kg War. War. 69 War. 120 War. 171 War. 222 War. 273 War. 324 War. 375 Kg War. War. 70 War. 121 War. 172 War. 223 War. 274 War. 325 War. 376 Kg War. War. 71 War. 122 War. 173 War. 224 War. 275 War. 326 War. 377 Kg War. 21 War. 72 War. 123 War. 174 War. 225 War. 276 War. 327 War. 378 Kg War. 22 War. 73 War. 124 War. 175 War. 226 War. 277 War. 328 War. 379 Kg War. 23 War. 74 War. 125 War. 176 War. 227 War. 278 War. 329 War. 380 Kg War. 24 War. 75 War. 126 War. 177 War. 228 War. 279 War. 330 War. 381 Kg War. 25 War. 76 War. 127 War. 178 War. 229 War. 280 War. 331 War. 382 Kg War. 26 War. 77 War. 128 War. 179 War. 230 War. 281 War. 332 War. 383 Kg War. 27 War. 78 War. 129 War. 180 War. 231 War. 282 War. 333 War. 384 Kg War. 28 War. 79 War. 130 War. 181 War. 232 War. 283 War. 334 War. 385 Kg War. 29 War. 8O War. 131 War. 182 War. 233 War. 284 War. 335 War. 386 22.5 Kg War. 30 War. 81 War. 132 War. 183 War. 234 War. 285 War. 336 War 387 US 2017/0044244 A1 Feb. 16, 2017 20

TABLE 1-continued Exemplary combinations of dosage and frequency for methods of treating a CNS disorder by intranasal administration of pooled human IgG. Two Three Four One Two Three Every One Time Times Times Times Time Times Times Other Monthly Monthly Monthly Monthly Weekly Weekly Weekly Day 25 mg/kg Var. 31 War. 82 War. 133 War. 184 War. 235 War. 286 War. 337 War. 27.5 mg/kg Var. 32 War. 83 War. 134 War. 185 War. 236 War. 287 War. 338 as 30 mg/kg Var. 33 War. 84 War. 135 War. 186 War. 237 War. 288 War. 339 3. 32.5 mg/kg Var. 34 War. 85 War. 136 War. 187 War. 238 War. 289 War. 340 2. 35 mg/kg Var. 35 War. 86 War. 137 War. 188 War. 239 War. 290 War. 341 R 37.5 mg/kg Var. 36 War. 87 War. 138 War. 189 War. 240 War. 291 War. 342 2. 40 mg/kg Var. 37 War. 88 War. 139 War. 190 War. 241 War. 292 War. 343 2. 45 mg/kg Var. 38 War. 89 War. 140 War. 191 War. 242 War. 293 War. 344 3. 50 mg/kg Var. 39 War. 90 War. 141 War. 192 War. 243 War. 294 War. 345 2. 0.5-40 mg/kg Var. 40 War. 91 War. 142 War. 193 War. 244 War. 295 War. 346 2. 0.5-30 mg/kg Var. 41 War. 92 War. 143 War. 194 War. 245 War. 296 War. 347 R 0.5-20 mg/kg Var. 42 War. 93 War. 144 War. 195 War. 246 War. 297 War. 348 3. 0.5-20 mg/kg Var. 43 War. 94 War. 145 War. 196 War. 247 War. 298 War. 349 2. 0.5-10 mg/kg Var. 44 War. 95 War. 146 War. 197 War. 248 War. 299 War. 350 s 0.5-5 mg/kg Var. 45 War. 96 War. 147 War. 198 War. 249 War. 300 War. 351 E.

1-20 mg/kg Var. 46 War. 97 War. 148 War. 199 War. 250 War. 301 War. 352

1-15 mg/kg Var. 47 War. 98 War. 149 War. 200 War. 251 War. 3O2 War. 353

1-10 mg/kg Var. 48 War. 99 War. 150 War. 201 War. 252 War. 303 War. 354

1-5 mg/kg Var. 49 War. 100 War. 151 War. 202 War. 253 War. 304 War. 355

2-10 mg/kg Var. 50 War. 101 War. 152 War. 203 War. 254 War. 305 War. 356 2-5 mg/kg Var. 51 War. 102 War. 153 War. 204 War. 255 War. 306 War. 357 s 408

War. = wariation

TABLE 2 Exemplary combinations of dosage and frequency for methods of treating a CNS disorder by intranasal administration of pooled human IgG. Four One Two Three Four Five Times Five Times Six Times Time Times Times Times Times Weekly Weekly Weekly Daily Daily Daily Daily Daily 0.05 mg/kg Var. 409 Var. 460 War. 511 War. 562 War. 613 War. 664 War. 715 War. 766 0.1 mg/kg Var. 410 Var. 461 War. 512 War. 563 War. 614 War. 665 War. 716 War. 767 0.25 mg/kg Var. 411 Var. 462 War. 513 War. 564 War. 615 War. 666 War. 717 War. 768 0.5 mg/kg Var. 412 Var. 463 War. 514 War. 565 War. 616 War. 667 War. 718 War. 769 0.75 mg/kg Var. 413 Var. 464 War. S15 War. S66 War. 617 War. 668 War. 719 War. 770 1.0 mg/kg Var. 414 Var. 465 War. S16 War. S67 War. 618 War. 669 War. 720 War. 771 1.5 mg/kg Var. 415 Var. 466 War. S17 War. S68 War. 619 War. 670 War. 721 War. 772 2.0 mg/kg Var. 416 Var. 467 War. S18 War. S69 War. 62O War. 671 War. 722 War. 773 2.5 mg/kg Var. 417 Var. 468 War. S19 War. S70 War. 621 War. 672 War. 723 War. 774 3.0 mg/kg Var. 418 Var. 469 War. S2O War. S71 War. 622 War. 673 War. 724 War. 775 3.5 mg/kg Var. 419 Var. 470 War. S21 War. S72 War. 623 War. 674 War. 725 War. 776 4.0 mg/kg Var. 420 Var. 471 War. S22 War. S73 War. 624 War. 675 War. 726 War. 777 4.5 mg/kg Var. 421 Var. 472 War. S23 War. S74 War. 62S War. 676 War. 727 War. 778 5.0 mg/kg Var. 422 Var. 473 War. S24 War. S75 War. 626 War. 677 War. 728 War. 779 US 2017/0044244 A1 Feb. 16, 2017 21

TABLE 2-continued Exemplary combinations of dosage and frequency for methods of treating a CNS disorder by intranasal administration of pooled human IgG. Four One Two Three Four Five Times Five Times Six Times Time Times Times Times Times Weekly Weekly Weekly Daily Daily Daily Daily Daily 6.0 mg/kg Var. 423 Var. 474 War. S2S War. S76 War. 627 War. 678 War. 729 War. 780 7.0 mg/kg Var. 424 Var. 475 War. S26 War. S77 War. 628 War. 679 War. 730 War. 781 8.0 mg/kg Var. 425 Var. 476 War. S27 War. S78 War. 629 War. 680 War. 731 War. 782 9.0 mg/kg Var. 426 Var. 477 War. S28 War. S79 War. 630 War. 681 War. 732 War. 783 10 mg/kg Var. 427 Var. 478 War. 529 War. 580 War. 631 War. 682 War. 733 War. 784 11 mg/kg Var. 428 Var. 479 War. S30 War. S81 War. 632 War. 683 War. 734 War. 785 12 mg/kg Var. 429 Var. 480 War. S31 War. 582 War. 633 War. 684 War. 735 War. 786 13 mg/kg Var. 430 Var. 481 War. 532 War. 583 War. 634 War. 68S War. 736 War. 787 14 mg/kg Var. 431 Var. 482 War. S33 War. S84 War. 635 War. 686 War. 737 War. 788 15 mg/kg Var. 432 Var. 483 War. S34 War. S85 War. 636 War. 687 War. 738 War. 789 16 mg/kg Var. 433 Var. 484 War. S35 War. S86 War. 637 War. 688 War. 739 War. 790 17 mg/kg Var. 434 Var. 485 War. S36 War. S87 War. 638 War. 689 War. 740 War. 791 18 mg/kg Var. 435 Var. 486 War. S37 War. S88 War. 639 War. 690 War. 741 War. 792 19 mg/kg Var. 436 Var. 487 War. 538 War. 589 War. 640 War. 691 War. 742 War. 793 20 mg/kg Var. 437 Var. 488 War. 539 War. 590 War. 641 War. 692 War. 743 War. 794 22.5 mg/kg Var. 438 Var. 489 War. 540 War. 591 War. 642 War. 693 War. 744 War. 795 25 mg/kg Var. 439 Var. 490 War. 541 War. 592 War. 643 War. 694 War. 745 War. 796 27.5 mg/kg Var. 440 War. 491 War. 542 War. 593 War. 644 War. 695 War. 746 War. 797 30 mg/kg Var. 441 Var. 492 War. 543 War. 594 War. 645 War. 696 War. 747 War. 798 32.5 mg/kg Var. 442 Var. 493 War. 544 War. 595 War. 646 War. 697 War. 748 War. 799 35 mg/kg Var. 443 Var. 494 War. 545 War. 596 War. 647 War. 698 War. 749 War. 800 37.5 mg/kg Var. 444 Var. 495 War. 546 War. 597 War. 648 War. 699 War. 7SO War. 801 40 mg/kg Var. 445 Var. 496 War. 547 War. 598 War. 649 War. 700 War. 751 War. 802 45 mg/kg Var. 446 Var. 497 War. 548 War. 599 War. 650 War. 701 War. 752 War. 803 50 mg/kg Var. 447 Var. 498 War. 549 War. 600 War. 651 War. 702 War. 753 War. 804 0.5-40 mg/kg Var. 448 Var. 499 War. SSO War. 601 War. 652 War. 703 War. 754 War. 805 0.5-30 mg/kg Var. 449 Var. 500 War. SS1 War. 602 War. 653 War. 704 War. 7SS War. 806 0.5-20 mg/kg Var. 450 Var. 501 War. 552 War. 603 War. 654 War. 70S War. 756 War. 807 0.5-20 mg/kg Var. 451 Var. 502 War. SS3 War. 604 War. 655 War. 706 War. 7S7 War. 808 0.5-10 mg/kg Var. 452 Var. 503 War. SS4 War. 60S War. 656 War. 707 War. 758 War. 809 0.5-5 mg/kg Var. 453 Var. 504 War. SSS War. 606 War. 657 War. 708 War. 759 War. 810 1-20 mg/kg Var. 454 Var. 505 War. SS6 War. 607 War. 658 War. 709 War. 760 War. 811 1-15 mg/kg Var. 455 Var. 506 War. SS7 War. 608 War. 659 War. 710 War. 761 War. 812 1-10 mg/kg Var. 456 Var. 507 War. 558 War. 609 War. 660 War. 711 War. 762 War. 813 1-5 mg/kg Var. 457 Var. 508 War. 559 War. 610 War. 661 War. 712 War. 763 War. 814 2-10 mg/kg Var. 458 War. 509 War. 560 War. 611 War. 662 War. 713 War. 764 War. 815 2-5 mg/kg Var. 459 Var. 510 War. 561 War. 612 War. 663 War. 714 War. 765 War. 816

War. = wariation

0206. Formulation for the subjects to be treated; each unit contains a predeter 0207 Pharmaceutical compositions of pooled human mined quantity of active compound calculated to produce immunoglobulin G described herein can be prepared in the desired therapeutic effect in association with the required accordance with methods well known and routinely prac pharmaceutical carrier. ticed in the art. See, e.g., Remington: The Science and 0208 Actual dosage levels can be varied so as to obtain Practice of Pharmacy, Mack Publishing Co., 20' ed., 2000; an amount of the active ingredient which is effective to and Sustained and Controlled Release Drug Delivery Sys achieve the desired therapeutic response for a particular tems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, patient without being toxic to the patient. A physician can 1978. Pharmaceutical compositions are preferably manufac start doses of the pharmaceutical composition at levels lower tured under GMP conditions. Typically, a therapeutically than that required to achieve the desired therapeutic effect effective dose or efficacious dose of the pooled human IgG and gradually increase the dosage until the desired effect is preparation is employed in the pharmaceutical compositions achieved. In general, effective doses vary depending upon described herein. The pharmaceutical composition can be many different factors, including the specific disease or formulated into dosage forms by conventional methods condition to be treated, its severity, physiological State of the known to those of skill in the art. Dosage regimens are patient, other medications administered, and whether treat adjusted to provide the optimum desired response (e.g., a ment is prophylactic or therapeutic. therapeutic response). For example, a single bolus may be 0209. In one embodiment, a therapeutic composition of administered, several divided doses may be administered pooled human IgG formulated for intranasal administration over time or the dose may be proportionally reduced or does not contain a permeability enhancer. Permeability increased as indicated by the exigencies of the therapeutic enhancers facilitate the transport of molecules through the situation. It can be advantageous to formulate parenteral mucosa, including the mucous, and the nasal epithelium. compositions in dosage unit form for ease of administration Non-limiting examples of absorption enhancers include and uniformity of dosage. Dosage unit form as used herein mucoadhesives, ciliary beat inhibitors, mucous fluidizers, refers to physically discrete units Suited as unitary dosages membrane fluidizers, and tight junction modulators. Specific US 2017/0044244 A1 Feb. 16, 2017 22 non-limiting examples include bile salts, phospholipids, agent in the formulation will be at or about 5 mM, 10 mM, Sodium glycyrrhetinate, Sodium caprate, ammonium tartrate, 15 mM, 20 mM, 25 mM, 50 mM, 75 mM, 100 mM, 125 gamma. aminolevulinic acid, oxalic acid, malonic acid, mM, 150 mM, 175 mM, 200 mM, 225 mM, 250 mM, 275 Succinic acid, maleic acid, and oxaloacetic acid. mM, 300 mM, 325 mM, 350 mM, 375 mM, 400 mM, 425 0210. In addition to pooled human IgG, the pharmaceu mM, 450 mM, 475 mM, 500 mM or higher. In a specific tical compositions provided herein include one or more embodiment, the buffering agent is glycine. stabilizing agents. In a specific embodiment, the stabilizing 0215. In yet other embodiments, the concentration of the agent is a buffering agent Suitable for intranasal administra buffering agent (e.g., glycine, histidine, or proline) in for tion. Non-limiting examples of buffering agents suitable for mulation (or in the solution from which a dry powder formulating the pooled human IgG compositions provided composition is to be prepared) is from 50 mM to 500 mM, herein include an amino acid (e.g., glycine, histidine, or 100 mM to 500 mM, 200 mM to 500 mM, 250 mM to 500 proline) a salt (e.g., citrate, phosphate, acetate, glutamate, mM, 300 mM to 500 mM, 50 mM to 300 mM, 100 mM to tartrate, benzoate, lactate, gluconate, malate. Succinate, for 300 mM, 200 mM to 300 mM, or 225 mM to 275 mM. In mate, propionate, or carbonate), or any combination thereof yet other specific embodiments, the concentration of the adjusted to an appropriate pH. Generally, the buffering agent buffering agent (e.g., glycine, histidine, or proline) in for will be sufficient to maintain a suitable pH in the formulation mulations provided herein is 250+50 mM, 250+40 mM, for an extended period of time. In a particular embodiment, 250+30 mM, 250-25 mM, 250+20 mM, 250-15 mM, the buffering agent is sufficient to maintain a pH of 4 to 7.5. 250-10 mM, 250+5 mM, or 250 mM. In a specific embodiment, the buffering agent is sufficient to 0216. In some embodiments, the pooled human immu maintain a pH of approximately 4.0, or approximately 4.5, noglobulins are formulated with between 100 mM and 400 or approximately 5.0, or approximately 5.5, or approxi mM histidine; no more than 10 mM of an alkali metal cation; mately 6.0, or approximately 6.5, or approximately 7.0, or and a pH between 5.0 and 7.0. approximately 7.5. 0217. In some embodiments of the pooled human immu 0211. In a particular embodiment, a pooled human IgG noglobulin histidine formulation, the concentration of his composition described herein for the treatment of a CNS tidine is between 5 mM and 500 mM. In another embodi disorder via intranasal administration contains a stabilizing ment, the concentration of histidine in the formulation will amount of an amino acid. In certain embodiments, a stabi be between 100 mM and 400 mM. In another embodiment, lizing amount of an amino acid is from about 25 mM to the concentration of histidine in the formulation will be about 500 mM between 200 mM and 300 mM. In another embodiment, the 0212. In a particular embodiment, the stabilizing agent concentration of histidine in the formulation will be between employed in the pooled human IgG compositions provided 225 mM and 275 mM. In another embodiment, the concen herein is an amino acid. Non-limiting examples of amino tration of histidine in the formulation will be between 240 acids include isoleucine, alanine, leucine, asparagine, lysine, mM and 260 mM. In a particular embodiment, the concen aspartic acid, methionine, cysteine, phenylalanine, glutamic tration of histidine will be 250 mM. In certain other embodi acid, threonine, glutamine, tryptophan, glycine, Valine, pro ments, the concentration of histidine in the formulation will line, selenocysteine, serine, tyrosine, arginine, histidine, be 5+0.5 mM, 10-1 mM, 15+1.5 mM, 20+2 mM, 25+2.5 ornithine, taurine, combinations thereof, and the like. In one mM, 50.5 mM, 75+7.5 mM, 100-10 mM, 125-12.5 mM, embodiment, the stabilizing amino acids include arginine, 150-15 mM, 175-17.5 mM, 200+20 mM, 225-22.5 mM, histidine, lysine, serine, proline, glycine, alanine, threonine, 250-25 mM, 275-27.5 mM, 300+30 mM, 325-32.5 mM, and a combination thereof. In a preferred embodiment, the 350-35 mM, 375+37.5 mM, 400-40 mM, 425-42.5 mM, amino acid is glycine. In another preferred embodiment, the 450+45 mM, 475+47.5 mM, 500+50 mM or higher. In yet amino acid is proline. In yet another preferred embodiment, other embodiments, the concentration of histidine in the the amino acid is histidine. formulation will be 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 0213 For purposes of stabilizing the compositions pro 50 mM, 75 mM, 100 mM, 125 mM, 150 mM, 175 mM, 200 vided herein, the buffering agent (e.g., glycine, histidine, or mM, 225 mM, 250 mM, 275 mM, 300 mM, 325 mM, 350 proline) will typically be added to the formulation (or to a mM, 375 mM, 400 mM, 425 mM, 450 mM, 475 mM, 500 solution from which a dry powder composition is to be mM or higher. prepared) at a concentration from 5 mM to 0.75 M. In one 0218. In some embodiments of the pooled human immu embodiment, at least 100 mM of the buffering agent is added noglobulin histidine formulation, the pH of the histidine to the formulation. In another embodiment, at least 200 mM formulation is from 4.0 to 7.5. In some embodiments, the pH of the buffering agent is added to the formulation. In yet of the histidine formulation is from 4.0 to 6.0. In some another embodiment, at least 250 mM of the buffering agent embodiments, the pH of the histidine formulation is from 4.0 is added to the formulation. In yet other embodiments, the to 4.5. In some embodiments, the pH of the histidine formulations provided herein contains at least 25 mM, 50 formulation is from 4.5 to 5.0. In some embodiments, the pH mM, 75 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 of the histidine formulation is from 4.0 to 5.5. In some mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 embodiments, the pH of the histidine formulation is from 4.0 mM, 650 mM, 700 mM, 750 mM, or more of the buffering to 6.5. In some embodiments, the pH of the histidine agent. In a specific embodiment, the buffering agent is formulation is from 4.0 to 7.0. In some embodiments, the pH glycine. of the histidine formulation is from 4.5 to 6.0. In some 0214. In one embodiment, the concentration of buffering embodiments, the pH of the histidine formulation is from 4.5 agent (e.g., glycine, histidine, or proline) in the formulation to 6.5. In some embodiments, the pH of the histidine (or in the solution from which a dry powder composition is formulation is from 4.5 to 7.0. In some embodiments, the pH to be prepared) is at or about from 5 mM to 500 mM. In of the histidine formulation is from 4.5 to 7.5. In some certain embodiments, the concentration of the buffering embodiments, the pH of the histidine formulation is from 5.5 US 2017/0044244 A1 Feb. 16, 2017

to 7.0. In some embodiments, the pH of the histidine g/L, 50 g/L to 250 g/L, 60 g/L to 250 g/L, 70 g/L to 250 g/L, formulation is from 6.0 to 7.0. In some embodiments, the pH 80 g/L to 250 g/L, 90 g/L to 250 g/L. 100 g/L to 250 g/L, 125 of the histidine formulation is from 6.5 to 7.0. In some g/L to 250 g/L, 150 g to 250 g/L, 175 g/L to 250 g/L, 200 embodiments, the pH of the histidine formulation is from 5.0 g/L to 250 g/L IgG. to 6.5. In some embodiments, the pH of the histidine 0222. In certain embodiments, the methods for treating a formulation is from 5.0 to 7.0. In some embodiments, the pH CNS disorder provided herein include intranasally admin of the histidine formulation is from 5.5 to 6.5. In some istering a liquid composition containing a low concentration embodiments, the pH of the histidine formulation is from 6.0 of pooled human IgG. In one embodiment, a low concen to 6.5. In some embodiments, the pH of the histidine tration of pooled human IgG contains from 1.0 g/L to 100 formulation is from 5.0 to 6.0. In some embodiments, the pH g/L, 5.0 g/L to 100 g/L. 10 g/L to 100 g/L, 20 g/L to 100 g/L, of the histidine formulation is from 5.5 to 6.0. In some 30g/L to 100 g/L, 40 g/L to 100 g/L, 50 g/L to 100 g/L, 60 embodiments, the pH of the histidine formulation is from 5.0 g/L to 100 g/L, 70 g/L to 100 g/L, 75 g/L to 100 g/L, 80 g/L to 5.5. In some embodiments, the pH of the histidine to 100 g/L, 1.0 g/L to 50 g/L, 5.0 g/L to 50 g/L. 10 g/L to formulation is from 7.0 to 7.5. In some embodiments, the pH 50 g/L, 20 g/L to 50 g/L, 30 g/L to 50 g/L, or 40 g/L to 50 of the histidine formulation is from 6.0 to 7.5. In some g/L IgG. embodiments, the pH of the histidine formulation is from 5.5 0223) In certain embodiments, the methods for treating a to 7.5. In some embodiments, the pH of the histidine CNS disorder provided herein include intranasally admin formulation is from 5.0 to 7.5. In some embodiments, the pH istering a liquid composition containing an intermediate of the histidine formulation is 5.0+0.2, 5.1+0.2, 5.2.0.2, concentration of pooled human IgG. In one embodiment, an 5.3+0.2, 5.4.0.2, 5.5.0.2, 5.6+0.2, 5.7.0.2, 5.8.0.2, 5.9-0. intermediate concentration of pooled human IgG contains 2, 6.0+0.2, 6.1+0.2, 6.2+0.2, 6.3-0.2, 6.4+0.2, 6.5+0.2, from 75 g/L to 200 g/L, 100 g/L to 200 g/L, 110 g/L to 200 6.6+0.2, 6.7+0.2, 6.8+0.2, 6.9-0.2, or 7.0+0.2. In some g/L, 120 g/L to 200 g/L, 130 g/L to 200 g/L, 140 g/L to 200 embodiments, the pH of the histidine formulation is 5.0+0.1, g/L, 150 g/L to 200 g/L, 160 g/L to 200 g/L, 170 g/L to 200 5.1+0.1, 5.2+0.1, 5.3+0.1, 5.4+0.1, 5.5+0.1, 5.6+0.1, 5.7+0. g/L, 175 g/L to 200 g/L, 180 g/L to 200 g/L, 75 g/L to 150 1, 5.8+0.1, 5.9+0.1, 6.0+0.1, 6.1+0.1, 6.2+0.1, 6.3-0.1, g/L. 100 g/L to 150 g/L, 110 g/L to 150 g/L, 120 g/L to 150 6.4+0.1, 6.5+0.1, 6.6+0.1, 6.7+0.1, 6.8+0.1, 6.9+0.1, or g/L, 130 g/L to 150 g/L, or 140 g/L to 150 g/L IgG. 7.0.0.1 0224. In certain embodiments, the methods for treating a 0219. In one embodiment, the pooled human IgG com CNS disorder provided herein include intranasally admin positions described herein for the treatment of a CNS istering a liquid composition containing a high concentration disorder via intranasal administration is formulated at a pH of pooled human IgG. In one embodiment, a high concen from about 4.0 to about 7.0. In particular embodiments, a tration of pooled human IgG contains from 175 g/L to 250 pooled human IgG compositions is formulated at a pH of g/L, 200 g/L to 250 g/L, 210 g/L to 250 g/L, 220 g/L to 250 about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, g/L, 230 g/L to 250 g/L, or 240 g/L to 250 g/L IgG. 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4., 6.5, 0225. In a particular embodiment, a liquid compositions 6.7, 6.8, 6.9, or 7.0. In other embodiments, a pooled human of pooled human IgG formulated for intranasal administra IgG composition is formulated at a pH from 4.0 to 6.5, 4.0 tion consists essentially of from 100 g/L to 250 g/L pooled to 6.0, 4.0 to 5.5, 4.0 to 5.0, 4.0 to 4.5, 4.5 to 6.5, 4.5 to 6.0, human IgG and from 150 mM to 350 mM glycine. 4.5 to 5.5, 4.5 to 5.0. In yet other embodiments, a pooled 0226. In another particular embodiment, a liquid compo human IgG composition is formulated at a pH of 4.8+0.5, sitions of pooled human IgG formulated for intranasal 4.8+0.4, 4.8+0.3, 4.8+0.2, 4.8+0.1, or about 4.8. administration consists essentially of from 150 g/L to 250 0220. In one embodiment, liquid compositions of pooled g/L pooled human IgG and from 200 mM to 300 mM human IgG formulated for intranasal administration are glycine. provided for the treatment of CNS disorders (e.g., Alzheim 0227. In yet another particular embodiment, a liquid er's disease, Parkinson's disease, and multiple Sclerosis). In compositions of pooled human IgG formulated for intranasal a specific embodiment, the liquid composition is an aqueous administration consists essentially of from 200 g/L to 250 composition. In a particular embodiment, an aqueous thera g/L pooled human IgG and 250+25 mM glycine. peutic composition formulated for intranasal administration 0228. In certain embodiments, the liquid compositions of provided herein consists essentially of a buffering agent and pooled human IgG formulated for intranasal administration pooled human IgG. provided herein further include a humectant. Non-limiting 0221. In one embodiment, a liquid composition formu examples of humectants include glycerin, polysaccharides, lated for intranasal administration contains from about 1.0 g and polyethylene glycols. pooled human IgG per liter (g/L IgG) to about 250 g/L IgG. 0229. In certain embodiments, the liquid compositions of In other embodiments, the liquid composition formulated for pooled human IgG formulated for intranasal administration intranasal administration contains about 1 g/L, 2 g/L, 3 g/L. provided herein further include an agent that increases the 4 g/L, 5g/L, 6 g/L, 7 g/L, 8 g/L, 9 g/L. 10 g/L, 12.5g/L, 15 flow properties of the composition. Non-limiting examples g/L, 17.5g/L, 20 g/L, 25g/L, 30 g/L, 35 g/L, 40 g/L, 45 g/L. of agents that increase to flow properties of an aqueous 50 g/L, 55 g/L, 60 g/L, 65 g/L, 70 g/L, 75 g/L, 80 g/L, 85 composition include Sodium carboxymethyl cellulose, g/L, 90 g/L. 95 g/L. 100 g/L, 110 g/L, 120 g/L, 130 g/L, 140 hyaluronic acid, gelatin, algin, carageenans, carbomers, g/L, 150 g/L, 160 g/L, 170 g/L, 180 g/L, 190 g/L, 200 g/L, galactomannans, polyethylene glycols, polyvinyl alcohol, 210 g/L, 220 g/L, 230 g/L, 240 g/L, 250 g/L, or higher polyvinylpyrrolidone, sodium carboxymethyl dextran, and concentration of pooled human IgG. In certain embodi Xantham. ments, the liquid composition formulated for intranasal 0230. In one embodiment, dry powder compositions of administration contains from 5.0 g/L to 250 g/L. 10 g/L to pooled human IgG formulated for intranasal administration 250 g/L, 20 g/L to 250 g/L, 30 g/L to 250 g/L, 40 g/L to 250 are provided for the treatment of CNS disorders (e.g., US 2017/0044244 A1 Feb. 16, 2017 24

Alzheimer's disease, Parkinson's disease, and multiple scle about 250 um. In some embodiments, the dry particles have rosis). In a specific embodiment, a dry powder therapeutic an average diameter between from 1 um to about 25um. In composition formulated for intranasal administration pro Some embodiments, the dry particles have an average diam vided herein consists essentially of a buffering agent and eter between from 10 um to about 100 um. In yet other pooled human IgG. embodiments, the dray particles have an average diameter of 0231. In one embodiment, a dry powder composition of about 0.1 um+10%, 0.2 um+10%, 0.3 um+10%, 0.4 pooled human IgG formulated for intranasal administration um+10%, 0.5 um+10%, 0.6 um+10%, 0.7 um+10%, 0.8 further comprises a bulking agent. Non-limiting examples of um+10%, 0.9 um+10%, 1.0 um+10%, 2 um+10%, 3 bulking agents include oxyethylene maleic anhydride copo um+10%, 4 um+10%, 5um+10%. 6 Limit 10%, 7 um+10%, lymer, polyvinylether, polyvinylpyrrolidone polyvinyl alco 8 um+10%, 9 um+10%, 10 um+10%, 11 um+10%, 12 hol, polyacrylates, including Sodium, potassium or ammo um+10%, 13 um+10%, 14 um+10%, 15 um+10%, 16 nium polyacrylate, polylactic acid, polyglycolic acid, um+10%, 17 um+10%, 18 umi-10%, 19 um+10%, 20 polyvinyl alcohol, polyvinyl acetate, carboxyvinyl polymer, um+10%, 25 um+10%, 30 um+10%, 35 um+10%, 40 polyvinylpyrrolidone, polyethylene glycol, celluloses (in um+10%, 45 um+10%, 50 um+10%, 60 um+10%. 65 cluding cellulose, microcrystalline cellulose, and alpha um+10%, 70 um+10%, 75 um+10%, 80 um+10%, 85 cellulose), cellulose derivatives (including methyl cellulose, um+10%, 90 um+10%, 95 um+10%, 100 um+10%, 110 ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cel um+10%, 120 um+10%, 130 um+10%, 140 um+10%, 150 lulose, hydroxypropyl methyl cellulose, sodium carboxym um+10%, 160 um+10%, 170 um+10%, 180 um+10%, 190 ethyl cellulose and ethylhydroxy ethyl cellulose), dextrins um+10%, 200 um+10%, 225 um+10%, 250 um+10%, 275 (including alpha-, beta-, or gamma-cyclodeXthn, and dim um+10%, 300 um+10%, 350 um+10%, 400 um+10%, 450 ethyl-beta-cyclodexthn), starches (including hydroxyethyl um+10%, 500 um+10%, or greater. starch, hydroxypropyl Starch, carboxymethyl starch), poly 0236. In one embodiment, gel, cream, or ointment com saccharides (including dextran, dextrin and alginic acid, positions of pooled human IgG formulated for intranasal hyaluronic acid, and pectic acid), carbohydrates (such as administration are provided for the treatment of CNS dis mannitol, glucose, lactose, fructose, Sucrose, and amylose), orders (e.g., Alzheimer's disease, Parkinson's disease, and proteins (including casein, gelatin, chitin, and chitosan), multiple Sclerosis). In a specific embodiment, a gel, cream, gums (such as gum arabic, Xanthan gum, tragacanth gum, or ointment therapeutic composition formulated for intrana and glucomannan), phospholipids, and combinations sal administration provided herein consists essentially of a thereof. buffering agent and pooled human IgG. 0232. In certain embodiments, a dry powder composition 0237. In one embodiment, a gel, cream, or ointment of pooled human IgG formulated for intranasal administra composition of pooled human IgG formulated for intranasal tion further comprises a mucosal penetration enhancer. Non administration further comprises a carrier agent. Non-lim limiting examples of mucosal penetration enhancers are bile iting examples of carrier agents for gel and ointment com salts, fatty acids, Surfactants and alcohols. Specific non positions include natural or synthetic polymers such as limiting examples of mucosal penetration enhancers are hyaluronic acid, sodium alginate, gelatin, corn starch, gum Sodium cholate, sodium dodecyl Sulphate, Sodium deoxy tragacanth, methylcellulose, hydroxyethylcellulose, car cholate, taurodeoxycholate, Sodium glycocholate, dimethyl boxymethylcellulose, Xanthan gum, dextrin, carboxymeth sulfoxide or ethanol. ylstarch, polyvinyl alcohol, Sodium polyacrylate, methoxy 0233. In certain embodiments, a dry powder composition ethylene maleic anhydride copolymer, polyvinylether, of pooled human IgG formulated for intranasal administra polyvinylpyrrolidone, fats and oils such as beeswax, olive tion further comprises a dispersant. A dispersant is an agent oil, cacao butter, Sesame oil, soybean oil, camelia oil, that assists aerosolization of the IgG or the absorption of the peanut oil, beef fat, lard, and lanolin, white petrolatum, IgG in intranasal mucosal tissue, or both. Non-limiting paraffins, hydrocabon gel ointments, fatty acids such as examples of dispersants are a mucosal penetration enhancers Stearic acid, alcohols such as cetyl alcohol and Stearyl and Surfactants. alcohol, polyethylene glycol, water, and combinations 0234. In certain embodiments, a dry powder composition thereof. of pooled human IgG formulated for intranasal administra 0238. In certain embodiments, the pooled human immu tion further comprises a bioadhesive agent. Non-limiting noglobulins are co-formulated with one or more vasocon examples of bioadhesive agents include chitosan or cyclo strictor agents. When present, the vasoconstrictor agent dextrin. In certain embodiments, a dry powder composition reduces non-target exposure (e.g., systemic exposure) of the of pooled human IgG formulated for intranasal administra pooled human immunoglobulin, by reducing absorption of tion further comprises a filler. Non-limiting examples of the immunoglobulins into the blood, effectively increasing fillers include Sugars, including lactose, Sucrose, mannitol, the targeting of the immunoglobulin to the CNS (e.g., to the or Sorbitol; cellulose preparations such as: for example, brain). Methods for the co-formulation of other pharmaceu maize starch, wheat starch, rice starch, potato starch, gelatin, ticals and vasoconstrictors can be found in U.S. Patent gum tragacanth, methylcellulose, microcrystalline cellulose, Application Publication No. 2008/0305077, the content of hydroxypropylmethylcellulose, sodium carboxymethylcel which is expressly incorporated herein by reference in its lulose; or others such as: polyvinylpyrrolidone (PVP or entirety for all purposes. Non-limiting examples of vaso povidone) or calcium phosphate. constrictors that may be co-formulated with pooled human 0235. The particle size a dry powder composition of immunoglobulins in this fashion include tetrahydrozoline, pooled human IgG can be determined by standard methods methoxamine, phenylephrine, ephedrine, norepinephrine, in the art. For example, the particles can be screened or oxymetazoline, tetrahydrozoline, Xylometazoline, clonidine, filtered through a mesh sieve. In certain embodiments, the guanabenz, guanfacine, C.-methyldopa, arginine vasopres dry particles have an average diameter from about 0.1 um to sin, and pseudoephedrine. US 2017/0044244 A1 Feb. 16, 2017

0239 Disorders of the Central Nervous System episodic or paroxysmal disorder of the central nervous 0240 IVIG treatment has been used in the treatment of system, a paralytic syndrome of the central nervous system, CNS disorders. Specifically, IVIG has been studied or used a nerve, nerve root, or plexus disorder of the central nervous in the treatment of Multiple Sclerosis (MS), stiff-person system, an organic mental disorder, a mental or behavioral syndrome, Alzheimer's disease (AD), postpolio syndrome, disorder caused by psychoactive Substance use, a schizo narcolepsy, stroke, and fibromyalgia and other pain Syn phrenia, Schizotypal, or delusional disorder, a mood (affec dromes. Stangle 2008 (Therapeutic Advances in Neurologi tive) disorder, neurotic, stress-related, or Somatoform disor cal Disorders, 1(2): 115-124). der, a behavioral syndrome, an adult personality or behavior 0241 IVIG has also been used to treat neuromyelitis disorder, a psychological development disorder, or a child optica (NMO). NMO, also known as Devic's disease or onset behavioral or emotional disorder. In some embodi Devic's syndrome, is an autoimmune, inflammatory disor ments, the CNS disorder is selected from the group consist der of the optic nerves and spinal cord. For example, a 2 g/kg ing of Alzheimer's disease, Parkinson's disease, multiple induction dose of IVIG followed by 0.4-0.5 g/kg monthly Sclerosis, amyotrophic lateral Sclerosis (ALS), Huntington's maintenance doses of IVIG has been used to treat NMO. disease, cerebral palsy, bipolar disorder, Schizophrenia, or Awad et al. 2011 (Current Neuropharmacology, 9:417428). Pediatric acute-onset neuropyschiatric syndrome (PANS). In 0242 IVIG has also been used and studied for the treat some embodiments, the CNS disorder is selected from the ment of acute disseminated encephalomyelitis (ADEM). group consisting of Alzheimer's disease, Parkinson's dis ADEM is an immune mediated disease of the brain. Spe ease, multiple Sclerosis, Pediatric Autoimmune Neuropsy cifically, ADEM involves autoimmune demyelination and is chiatric Disorders Associated with Streptococcal infections classified as a MS borderline disease. For example, a stan (PANDAS), or Pediatric acute-onset neuropyschiatric syn dard dose of 2 g/kg IVIG given over 2-5 days can be used drome (PANS). to treat ADEM. Pohl et al. 2012 (Current Treatment Options 0247. In one embodiment, the CNS disorder is a systemic in Neurology, 14:264-275). atrophy primarily affecting the central nervous system. Non 0243 IVIG has also studied and used in the treatment of limiting examples of systemic atrophies that primarily affect Parkinson's disease (PD). For example, studies have shown the central nervous system include: Huntington's disease; that IVIG may reduce C-synuclein neurotoxicity, a possible hereditary ataxias (e.g., congenital non-progressive ataxia, contributing factor to the pathogenesis of PD, through an early-onset cerebellar ataxias—such as early-onset cerebel unknown mechanism. Smith et al. 2012 (International lar ataxia with essential tremor, Hunt's ataxia, early-onset Immunopharmacology, 14:550-557) and Patrias et al. (Clini cerebellar ataxia with retained tendon reflexes, Friedreich's cal and Experimental Immunology, 161:527-535). ataxia, and X-linked recessive spinocerebellar ataxia—late 0244 IVIG has also been used and studied for the treat onset cerebellar ataxia, ataxia telangiectasia (Louis-Bar Syn ment of MS. For example, IVIG has been used successfully drome), or hereditary spastic paraplegia); a spinal muscular in the treatment of Schilder's disease (SD), a rare variant of atrophy or related disorder thereof (e.g., Werdnig-Hoffman MS. Krause et al. 2012 (European J. of Paediatric Neurol disease (Type 1), progressive bulbar palsy of childhood ogy, 16:206-208). IVIG has also been suggested to be (Fazio-Londe syndrome), Kugelberg-Welander disease beneficial in the treatment of acute relapses in MS patients. (Type 3), or a motor neuron disease—such as familial motor Elovaara et al. 2011 (Clinical Neuropharmacology, 34(2): neuron disease, amyotrophic lateral Sclerosis (ALS), pri 84-89). mary lateral Sclerosis, progressive bulbar palsy, and progres 0245 IVIG has also been used and studied for the treat sive spinal muscular atrophy); paraneoplastic neuromyopa ment of obsessive-compulsive disorders (OCD) and tic thy and neuropathy; Systemic atrophy primarily affecting the disorders. For example, IVIG was shown to lessen the central nervous system in neoplastic disease; paraneoplastic severity of symptoms of OCD and tic disorders in children limbic encephalopathy; and systemic atrophy primarily with infection-triggered OCD and tic disorders. Perlmutter, affecting the central nervous system in myxoedema. et al. 1999 (The Lancet, 354:1153-1158). Similarly, it has 0248. In one embodiment, the CNS disorder is an been shown that IVIG is effective in reducing neuropsychi extrapyramidal and movement disorder. Non-limiting atric symptom severity in a subgroup OCD and tic disorder examples of extrapyramidal and movement disorders that patients with childhood-onset OCD and tic disorders. Snider affect the central nervous system include: Parkinson's dis et al. 2003 (J. of Child and Adolescent Psychopharmacology, ease; a secondary parkinsonism (e.g., malignant neuroleptic 13(supp 1): S81-S88). syndrome or postencephalitic parkinsonism); a degenerative 0246. In one aspect, the present invention provides a disease of the basal ganglia (e.g., Hallervorden-Spatz dis method for treating a central nervous system (CNS) disorder ease, progressive Supranuclear ophthalmoplegia (Steele in a subject in need thereof by delivering a therapeutically Richardson-Olszewski disease), or striatonigral degenera effective amount of a composition comprising pooled human tion), a dystonia (e.g., drug-induced dystonia, idiopathic immunoglobulin G (IgG) to the brain of the subject, wherein familial dystonia, idiopathic non-familial dystonia, Spas delivering the composition to the brain comprises intrana modic torticollis, idiopathic orofacial dystonia—such as Sally administering the composition directly to an epithelium orofacial dyskinesia—or blepharospasm); an essential of the nasal cavity of the Subject. In a specific embodiment, tremor; a drug-induced tremor, myoclonus, drug-induced the composition is administered directly to the olfactory chorea, drug-induced tics; restless legs syndrome; and stiff epithelium of the nasal cavity. In certain embodiments, the man syndrome. CNS disorder is selected from the group consisting of a 0249. In one embodiment, the CNS disorder is a neuro systemic atrophy primarily affecting the central nervous degenerative disorder of the central nervous system. Non system, an extrapyramidal and movement disorder, a neu limiting examples of neurodegenerative disorders that affect rodegenerative disorder of the central nervous system, a the central nervous system include: Alzheimer's disease; a demyelinating disorder of the central nervous system, an circumscribed brain atrophy (e.g., Pick's disease); senile US 2017/0044244 A1 Feb. 16, 2017 26 degeneration of brain; a degeneration of nervous system due middle cerebral artery syndrome, anterior cerebral artery to alcohol; grey-matter degeneration (e.g., Alpers disease); syndrome, posterior cerebral artery syndrome, a brain stem Lewy body dementia, Subacute necrotizing encephalopathy stroke syndrome (e.g., Benedikt Syndrome, Claude Syn (e.g., Leigh's disease); and Subacute combined degeneration drome, Foville syndrome, Millard-Gubler syndrome, Wal of spinal cord. In certain embodiments, the CNS disorder is lenberg syndrome, or Weber syndrome), cerebellar stroke disorder characterized by dementia. In certain embodiments, syndrome, pure motor lacunar syndrome, pure sensory lacu the dementia is a cortical dementia (associated, for example, nar syndrome, or a lacunar syndromes); and a sleep disorder with Alzheimer's) arising from a disorder affecting the (e.g., insomnia, hyperinsomnia, a disruption in circadian cerebral cortex. In certain embodiments, the dementia is a rhythm, sleep apnoea, narcolepsy, or cataplexy). Subcortical dementia (associated, for example, with Parkin 0252. In one embodiment, the CNS disorder is a paralytic son's disease and Huntington's disease) resulting from dys syndrome of the central nervous system. Non-limiting function in the parts of the brain that are beneath the cortex. examples of paralytic syndromes that affect the central In certain embodiments, the dementia is a side effect of drug nervous system include: a cerebral palsy (e.g., spastic quad administration. In specific embodiments, the dementia is a riplegic cerebral palsy, spastic diplegic cerebral palsy, spas side effect of the administration of a chemotherapeutic tic hemiplegic cerebral palsy, dyskinetic cerebral palsy, or agent. In specific embodiments, the dementia is a result of ataxic cerebral palsy); a hemiplegia (e.g., flaccid hemiplegia undergoing cardiac bypass. In specific embodiments, the or spastic hemiplegia); a paraplegia or tetraplegia (e.g., dementia is a result of a vascular disorder (e.g., myocardial flaccid paraplegia, spastic paraplegia, paralysis of both infarction, stroke, high blood pressure). In specific embodi lower limbs, lower paraplegia, flaccid tetraplegia, spastic ments, the dementia is a result of depression. tetraplegia, or quadriplegia); diplegia of upper limbs; 0250 In one embodiment, the CNS disorder is a demy monoplegia of a lower limb, monoplegia of an upper limb; elinating disorder of the central nervous system. Non-lim cauda equina syndrome; and Todd's paralysis (postepilep iting examples of demyelinating disorders that affect the tic). central nervous system include: multiple Sclerosis; an acute 0253. In one embodiment, the CNS disorder is a nerve, disseminated demyelination disorder (e.g., neuromyelitis nerve root, or plexus disorder of the central nervous system. optica (Devic's syndrome) or acute and Subacute hemor Non-limiting examples of nerve, nerve root, or plexus rhagic leukoencephalitis (Hurst's disease)); diffuse Sclero disorders that affect the central nervous system include: a sis; central demyelination of corpus callosum, central pon disorder of the trigeminal nerve (V; e.g., trigeminal neural tine myelinolysis; acute transverse myelitis in demyelinating gia); a facial nerve disorders (VII; e.g., bell's palsy, facial disease of central nervous system; Subacute necrotizing palsy, geniculate ganglionitis, melkersson’s syndrome, myelitis; and concentric sclerosis (Baló disease). melkersson-Rosenthal syndrome, a clonic hemifacial spasm, 0251. In one embodiment, the CNS disorder is an epi facial myokymia); a disorder of the olfactory nerve (I); a sodic or paroxysmal disorder of the central nervous system. disorder of the glossopharyngeal nerve (IX); a disorder of Non-limiting examples of episodic and paroxysmal disor the vagus nerve (X); a disorder of the hypoglossal nerve ders that affect the central nervous system include: epilepsy (XII); a disorder of multiple cranial nerves; and a nerve root (e.g., localization-related (focal) (partial) idiopathic epilepsy or plexus disorder affecting the CNS (e.g., a brachial plexus and epileptic syndromes with seizures of localized onset, disorder—such as thoracic outlet syndrome—a lumbosacral localization-related (focal)(partial) symptomatic epilepsy plexus disorder, a cervical root, a thoracic root disorder, a and epileptic syndromes with simple partial seizures; local lumbosacral root disorder, a neuralgic amyotrophy—such as ization-related (focal) (partial) symptomatic epilepsy and Parsonage-Aldren-Turner syndrome—or phantom limb Syn epileptic syndromes with complex partial seizures; a benign drome with or without pain). epileptic syndrome—such as myoclonic epilepsy in infancy 0254. In one embodiment, the CNS disorder is an other and neonatal convulsions (familial)—childhood absence wise classified disorder of the central nervous system. Non epilepsy (e.g., pyknolepsy), epilepsy with grand mal sei limiting examples of these disorders include: hydrocepha Zures on awakening, a juvenile epilepsy—Such as absence lus; a toxic encephalopathy, a cerebral cyst; anoxic brain epilepsy or myoclonic epilepsy (impulsive petit mal)—a damage; benign intracranial hypertension; postviral fatigue nonspecific epileptic seizure—such as an atonic, clonic, syndrome; an encephalopathy; compression of brain; cere myoclonic, tonic, or tonic-clonic epileptic seizure, epilepsy bral oedema; reye's syndrome; postradiation encephalopa with myoclonic absences or myoclonic-astatic seizures, thy; traumatic brain injury; Syringomyelia: Syringobulbia; a infantile spasms, Lennox-Gastaut syndrome, Salaam vascular myelopathy; spinal cord compression; myelopathy; attacks, symptomatic early myoclonic encephalopathy, a cerebrospinal fluid leak; a disorder of the meninges (e.g., West's syndrome, epilepsia partialis continua (Kozhevnikov cerebral or spinal meningeal adhesion); and a post-proce epilepsy), grand mal seizures, or petit mal); headaches (e.g., dural disorder of nervous system (e.g., cerebrospinal fluid a migraine—such as a migraine without aura (common leak from spinal puncture, an adverse reaction to a spinal or migraine), a migraine with aura (classical migraine), status lumbar puncture, or intracranial hypotension following ven migrainosus, and complicated migraine—cluster headache tricular shunting). syndrome, a vascular headache, a tension-type headache, a 0255. In one embodiment, the CNS disorder is an organic chronic post-traumatic headache, or a drug-induced head mental disorder. Non-limiting examples of organic mental ache); a cerebrovascular episodic or paroxysmal disorder disorders that affect the central nervous system include: (e.g., a transient cerebral ischaemic attacks or related Syn dementia (e.g., dementia associated with Alzheimer's dis drome—such as vertebrobasilar artery syndrome, carotid ease, Pick's disease, Creutzfeldt-Jakob disease, Hunting artery syndrome (hemispheric), a multiple and bilateral ton's disease, Parkinson's disease, or human immunodefi precerebral artery syndrome, amaurosis fugax, and transient ciency virus (HIV) disease, or vascular dementia—such as global amnesia—a vascular syndrome of the brain—such as multi-infarct dementia); organic amnesic syndrome not US 2017/0044244 A1 Feb. 16, 2017 27 induced by alcohol and other psychoactive Substances); affective disorder—current episode severe depression with delirium not induced by alcohol and other psychoactive psychotic symptoms, bipolar affective disorder—current Substances; a mental disorder due to brain damage and episode mixed, bipolar affective disorder—currently in dysfunction and to physical disease (e.g., organic halluci remission, bipolar II disorder, or recurrent manic episodes); nosis, organic catatonic disorder, organic delusional (schizo a depressive episode (e.g., mild depressive episode, moder phrenia-like) disorder, organic mood (affective) disorder, ate depressive episode, severe depressive episode without organic anxiety disorder, organic dissociative disorder; psychotic symptoms, severe depressive episode with psy organic emotionally labile (asthenic) disorder; a mild cog chotic symptoms, atypical depression, or single episodes of nitive disorder, or organic brain syndrome); and a person “masked depression); a recurrent depressive disorder (e.g., ality and behavioral disorders due to brain disease, damage and dysfunction (e.g., organic personality disorder, posten recurrent depressive disorder—current episode mild, recur cephalitic syndrome, or postconcussional syndrome). rent depressive disorder—current episode moderate, recur rent depressive disorder—current episode severe without 0256 In one embodiment, the CNS disorder is a mental psychotic symptoms, recurrent depressive disorder—current or behavioral disorder caused by psychoactive substance episode severe with psychotic symptoms, or recurrent use. Non-limiting examples of mental or behavioral disor depressive disorder—currently in remission); a persistent ders caused by psychoactive Substance use that affect the mood (affective) disorder (e.g., cyclothymia or dysthymia); central nervous system include: acute intoxication (e.g., mixed affective episode; and recurrent brief depressive from alcohol, opioid, cannabis, benzodiazepine, or cocaine episodes. use); a dependence syndrome (e.g., from alcohol, opioid, cannabis, benzodiazepine, cocaine, or nicotine addiction); a 0260. In one embodiment, the CNS disorder is a neurotic, withdrawal syndrome (e.g., an alcohol or benzodiazepine stress-related, or somatoform disorder. Non-limiting withdrawal syndrome); delirium tremens; and a psychotic examples of neurotic, stress-related, or somatoform disor disorder (e.g., alcoholic hallucinosis or stimulant psychosis); ders that affect the central nervous system include: a phobic an amnesic syndrome (e.g., Korsakoff S syndrome); a anxiety disorder (e.g., agoraphobia, anthropophobia, Social residual and late-onset psychotic disorder (e.g., posthalluci neurosis, acrophobia, animal phobias, claustrophobia, or nogen perception disorder). simple phobia); an otherwise categorized anxiety disorder 0257. In one embodiment, the CNS disorder is an autism (e.g., panic disorder (episodic paroxysmal anxiety) or gen spectrum disorder. In certain embodiments, the CNS disor eralized anxiety disorder); obsessive-compulsive disorder; der is autism, Asperger syndrome, pervasive developmental an adjustment disorder (e.g., acute stress reaction; post disorder not otherwise specified (PDD-NOS), childhood traumatic stress disorder, or adjustment disorder); a disso disintegrative disorder, or Rett syndrome. ciative (conversion) disorder (e.g., dissociative amnesia, 0258. In one embodiment, the CNS disorder is a schizo dissociative fugue, dissociative stupor; trance disorder, pos phrenia, Schizotypal, or delusional disorder. Non-limiting session disorder, dissociative motor disorder, dissociative examples of Schizophrenia, Schizotypal, and delusional dis convulsions, dissociative anaesthesia and sensory loss, orders that affect the central nervous system include: schizo mixed dissociative (conversion) disorder, Ganser's Syn phrenia (e.g., paranoid Schizophrenia, hebephrenic schizo drome, or multiple personality disorder); a Somatoform phrenia (disorganized Schizophrenia), catatonic disorder (e.g., Briquet's disorder, multiple psychosomatic Schizophrenia, undifferentiated Schizophrenia, post-Schizo disorder, a hypochondriacal disorder—such as body dys phrenic depression, residual schizophrenia, simple schizo morphic disorder, dysmorphophobia (nondelusional), hypo phrenia, cenesthopathic Schizophrenia, Schizophreniform chondriacal neurosis, hypochondriasis, and nosophobia—a disorder, or Schizophreniform psychosis); Schizotypal disor Somatoform autonomic dysfunction—such as cardiac neu der; a persistent delusional disorder (e.g., delusional disor rosis, Da Costa's syndrome, gastric neurosis, and neurocir der, delusional dysmorphophobia, involutional paranoid culatory asthenia—or psychalgia); neurasthenia; deperson state, or paranoia querulans); an acute or transient psychotic alization-derealization syndrome; Dhat syndrome, disorder (e.g., acute polymorphic psychotic disorder without occupational neurosis (e.g., writer's cramp); psychasthenia; symptoms of schizophrenia, acute polymorphic psychotic psychasthenic neurosis; and psychogenic syncope. disorder with symptoms of Schizophrenia, or acute schizo 0261. In one embodiment, the CNS disorder is a behav phrenia-like psychotic disorder); an induced delusional dis ioral syndrome associated with physiological disturbances order (e.g., folie a deux, induced paranoid disorder, or or physical factors. Non-limiting examples of behavioral induced psychotic disorder); a schizoaffective disorder (e.g., syndromes associated with physiological disturbances or manic type, depressive type, or mixed type schizoaffective physical factors that affect the central nervous system disorder); and chronic hallucinatory psychosis. include: an eating disorder (e.g., anorexia nervos, atypical 0259. In one embodiment, the CNS disorder is a mood anorexia nervosa, bulimia nervosa, atypical bulimia nervosa, (affective) disorder. Non-limiting examples of mood (affec overeating associated with other psychological disturbances, tive) disorders that affect the central nervous system include: Vomiting associated with other psychological disturbances, a manic episode (e.g., hypomania, mania without psychotic or pica in adults); a nonorganic sleep disorder (e.g., nonor symptoms, or mania with psychotic symptoms); a bipolar ganic insomnia, nonorganic hypersomnia, nonorganic dis affective disorder (e.g., bipolar affective disorder—current order of the sleep-wake schedule, sleepwalking (Somnam episode hypomanic, bipolar affective disorder—current epi bulism), sleep terrors (night terrors), or nightmares); a sode manic without psychotic symptoms, bipolar affective sexual dysfunction not caused by organic disorder or dis disorder—current episode manic with psychotic symptoms, ease; a mental or behavioral disorder associated with the bipolar affective disorder—current episode mild or moder puerperium (e.g., postnatal depression, postpartum depres ate depression, bipolar affective disorder—current episode Sion, or puerperal psychosis); and abuse of non-dependence severe depression without psychotic symptoms, bipolar producing Substances. US 2017/0044244 A1 Feb. 16, 2017 28

0262. In one embodiment, the CNS disorder is an adult of infancy and childhood, Stereotyped movement disorders, personality or behavior disorder. Non-limiting examples of Stuttering (stammering), cluttering, attention deficit disorder adult personality and behavior disorders that affect the without hyperactivity, Pediatric Autoimmune Neuropsychi central nervous system include: a specific personality dis atric Disorders Associated with Streptococcal infections order (e.g., paranoid personality disorder, Schizoid person (PANDAS), or Pediatric acute-onset neuropyschiatric syn ality disorder, a dissocial personality disorder—such as drome (PANS)). antisocial personality disorder—an emotionally unstable 0265. In one embodiment of the method for treating a personality disorder—such as borderline personality disor CNS disorder, the method includes intranasally administer der—histrionic personality disorder, an anankastic person ing a dry powder composition containing from 0.05 mg/kg ality disorder—such as obsessive-compulsive personality to 50 mg/kg pooled human immunoglobulin to a Subject in disorder, anxious (avoidant) personality disorder, dependent need thereof daily. In other embodiments, the methods personality disorder, eccentric personality disorder, haltlose provided herein for the treatment of a CNS disorder include personality disorder, immature personality disorder, narcis intranasally administering a dry powder composition of sistic personality disorder, passive-aggressive personality pooled human IgG in a dosage/frequency combination disorder, or psychoneurotic personality disorder); mixed selected from variations 1 to 816 found in Table 1 and Table personality disorder, a habit or impulse disorder (e.g., patho 2. In a particular embodiment, the method comprises admin logical gambling, pathological fire-setting (pyromania), istering the dry powder composition directly to a nasal pathological stealing (kleptomania), or trichotillomania); epithelium of the subject. In a particular embodiment, the and Munchausen syndrome. method comprises administering the dry powder composi 0263. In one embodiment, the CNS disorder is a psycho tion directly to the olfactory epithelium of the subject. logical development disorder. Non-limiting examples of 0266. In one embodiment of the method for treating a psychological development disorders that affect the central CNS disorder, the method includes intranasally administer nervous system include: a developmental disorder of speech ing a liquid (e.g., an aqueous) composition containing from or language (e.g., specific speech articulation disorder, 0.05 mg/kg to 50 mg/kg pooled human immunoglobulin to expressive language disorder, receptive language disorder a subject in need thereof daily. In other embodiments, the (receptive aphasia), acquired aphasia with epilepsy (Landau methods provided herein for the treatment of a CNS disorder Kleffner disorder), or lisping); a developmental disorder of include intranasally administering a liquid (e.g., an aqueous) scholastic skills (e.g., a specific reading disorder—such as composition of pooled human IgG in a dosage/frequency developmental dyslexia—specific spelling disorder, a spe combination selected from variations 1 to 816 found in Table cific disorder of arithmetical skills—such as developmental 1 and Table 2. In a particular embodiment, the method acalculia and Gerstmann syndrome—or a mixed disorder of comprises administering the composition drop-wise directly scholastic skills); a developmental disorder of motor func to a nasal epithelium of the Subject. In a particular embodi tion (e.g., developmental dyspraxia); a mixed specific devel ment, the method comprises administering the composition opmental disorder, and a pervasive developmental disorder drop-wise directly to the olfactory epithelium of the subject. (e.g., childhood autism, atypical autism, Rett's syndrome, In another particular embodiment, the method comprises overactive disorder associated with mental retardation and administering the composition via a spray directly to a nasal Stereotyped movements, or Asperger's syndrome). epithelium of the subject. In a particular embodiment, the 0264. In one embodiment, the CNS disorder is a behav method comprises administering the composition via a spray ioral or emotional disorder with onset usually occurring in directly to the olfactory epithelium of the subject. childhood and adolescence. Non-limiting examples of 0267 In one embodiment of the method for treating a behavioral or emotional disorders with onset usually occur CNS disorder, the method includes intranasally administer ring in childhood and adolescence that affect the central ing a gel, cream, or ointment composition containing from nervous system include: a hyperkinetic disorder (e.g., a 0.05 mg/kg to 50 mg/kg pooled human immunoglobulin to disturbance of activity and attention—such as attention a subject in need thereof daily. In other embodiments, the deficit hyperactivity disorder and attention deficit syndrome methods provided herein for the treatment of a CNS disorder with hyperactivity—or hyperkinetic conduct disorder); a include intranasally administering a gel, cream, or ointment conduct disorder (e.g., conduct disorder confined to the composition of pooled human IgG in a dosage/frequency family context, unsocialized conduct disorder, Socialized combination selected from variations 1 to 816 found in Table conduct disorder, or oppositional defiant disorder); a mixed 1 and Table 2. In a particular embodiment, the method disorder of conduct or emotions (e.g., depressive conduct comprises administering the gel, cream, or ointment com disorder); an emotional disorder with onset specific to position directly to a nasal epithelium of the Subject. In a childhood (e.g., separation anxiety disorder of childhood, particular embodiment, the method comprises administering phobic anxiety disorder of childhood, social anxiety disorder the gel, cream, or ointment composition directly to the of childhood, sibling rivalry disorder, identity disorder, or olfactory epithelium of the subject. overanxious disorder); a disorder of Social functioning with 0268 Alzheimer's Disease onset specific to childhood and adolescence (e.g., elective 0269. IVIG has been used in the treatment of Alzheimer's mutism, reactive attachment disorder of childhood, or dis disease. It has been proposed that IVIG contains antibodies inhibited attachment disorder of childhood); a tic disorder against B-amyloid. Relkin et al. 2009 (Neurobiol. Aging (e.g., transient tic disorder, chronic motor or vocal tic 30(11): 1728-36). In this study, pooled human IgG was disorder, or combined vocal and multiple motor tic disorder administered intravenously (IVIG therapy) to eight subjects (de la Tourette); and an otherwise classified behavioral or diagnosed with mild Alzheimer's disease (AD). The patients emotional disorder with onset usually occurring in child received IVIG therapy for 6 months, discontinued treatment, hood and adolescence (e.g., nonorganic enuresis, nonorganic and then resumed treatment for 9 more months. It was found encopresis, feeding disorder of infancy and childhood, pica that B-amyloid antibodies in the serum from AD patients US 2017/0044244 A1 Feb. 16, 2017 29 increased in proportion to IVIG dose and plasma levels of noglobulin to a subject in need thereof daily. In other B-amyloid increased transiently after each infusion. After 6 embodiments, the methods provided herein for the treatment months of treatment, mini-mental state tests were performed of Alzheimer's disease include intranasally administering a on the patients. The mini-mental state scores increased an gel, cream, or ointment composition of pooled human IgG in average of 2.5 points after 6 months, returned to baseline a dosage/frequency combination selected from variations 1 during washout and remained stable during Subsequent IVIG to 816 found in Table 1 and Table 2. In a particular treatment. embodiment, the method comprises administering the gel. 0270. In one aspect, the present invention provides a cream, or ointment composition directly to a nasal epithe method for treating Alzheimer's disease in a Subject in need lium of the subject. In a particular embodiment, the method thereof by delivering a therapeutically effective amount of a comprises administering the gel, cream, or ointment com composition comprising pooled human immunoglobulin G position directly to the olfactory epithelium of the subject. In (IgG) to the brain of the subject, wherein delivering the one embodiment, the Alzheimer's disease is early-onset composition to the brain comprises intranasally administer Alzheimer's disease. In another embodiment, the Alzheim ing the composition directly to an epithelium of the nasal er's disease is late-onset Alzheimer's disease. cavity of the Subject. In a specific embodiment, the compo (0274) Multiple Sclerosis sition is administered directly to the olfactory epithelium of 0275 Multiple sclerosis (MS) is a chronic neurodegen the nasal cavity. In one embodiment, the Alzheimer's dis erative and inflammatory disease of the central nervous ease is early-onset Alzheimer's disease. In another embodi system (CNS) that represents one of the most prevalent ment, the Alzheimer's disease is late-onset Alzheimer's human autoimmune diseases. Multiple Sclerosis (MS) is an disease. autoimmune disease that specifically affects the brain and 0271 In one embodiment of the method for treating spinal cord. MS is caused by damage to the myelin sheath, Alzheimer's disease, the method includes intranasally the protective covering that surrounds nerve cells. When the administering a dry powder composition containing from myelin sheath is damaged, nerve signals slow down or stop. 0.05 mg/kg to 50 mg/kg pooled human immunoglobulin to Damage to the myelin sheath is caused by inflammation a subject in need thereof daily. In other embodiments, the which occurs when the body's own immune cells attack the methods provided herein for the treatment of Alzheimer's nervous system. This can occur along any area of the brain, disease include intranasally administering a dry powder optic nerve, and spinal cord. composition of pooled human IgG in a dosage/frequency 0276 MS is classified into four subtypes based on the combination selected from variations 1 to 816 found in Table disease's progression: Relapsing-Remitting MS (RMSS), 1 and Table 2. In a particular embodiment, the method Secondary Progressive MS (SPMS), Primary-Progressive comprises administering the dry powder composition MS (PPMS), and Progressive-Relapsing MS (PRMS). More directly to a nasal epithelium of the Subject. In a particular than 80 percent of patients who are diagnosed with MS embodiment, the method comprises administering the dry exhibit initial signs of RMSS. RMSS is characterized by powder composition directly to the olfactory epithelium of relapse (characterized by symptom flare-ups) followed by the subject. In one embodiment, the Alzheimer's disease is remission. The relapses can be mild to severe flare-ups and early-onset Alzheimer's disease. In another embodiment, the the remissions can last for days to months. RMSS patients Alzheimer's disease is late-onset Alzheimer's disease. often develop SPMS. SPMS is characterized by relapses 0272. In one embodiment of the method for treating followed by only partial recoveries. During the partial Alzheimer's disease, the method includes intranasally recovery phase, the symptoms may lessen but do not go into administering a liquid (e.g., an aqueous) composition con full remission. SPMS is a progressive subtype of MS taining from 0.05 mg/kg to 50 mg/kg pooled human immu wherein the symptoms steadily worsen until a chronic noglobulin to a subject in need thereof daily. In other disability replaces the cycles of recovery and partial recov embodiments, the methods provided herein for the treatment ery. PPMS accounts for approximately 15 percent of MS of Alzheimer's disease include intranasally administering a occurrences. It is characterized by a slow and steady pro liquid (e.g., an aqueous) composition of pooled human IgG gression without periods of remission or partial recovery. in a dosage/frequency combination selected from variations PRMS is the least common subtype of MS. PRMS is 1 to 816 found in Table 1 and Table 2. In a particular characterized by steadily worsening symptoms and attacks embodiment, the method comprises administering the com followed by periods of remission. position drop-wise directly to a nasal epithelium of the 0277. There are peptide-induced and transgenic mouse Subject. In a particular embodiment, the method comprises model for MS. Experimental autoimmune encephalomyeli administering the composition drop-wise directly to the tis (EAE) is an animal model of brain inflammation. EAE is olfactory epithelium of the subject. In another particular an inflammatory demyelinating disease of the CNS. Acute embodiment, the method comprises administering the com and relapsing EAE is characterized by the formation of focal position via a spray directly to a nasal epithelium of the inflammatory demyelinating lesions in the white matter of Subject. In a particular embodiment, the method comprises the brain. This phenotype can be induced in normal SJL administering the composition via a spray directly to the mice through the administration of PLP139-151 peptide. olfactory epithelium of the subject. In one embodiment, the Chronic progressive EAE is pathologically associated with Alzheimer's disease is early-onset Alzheimer's disease. In a widespread axonal damage in the normal appearing white another embodiment, the Alzheimer's disease is late-onset matter and massive demyelination in the grey matter, par Alzheimer's disease. ticularly in the cortex. This phenotype can be induced in 0273. In one embodiment of the method for treating normal C57BL/6 mice through the administration of Alzheimer's disease, the method includes intranasally MOG35-55 peptide. administering a gel, cream, or ointment composition con 0278. There is also evidence that tumor necrosis factor taining from 0.05 mg/kg to 50 mg/kg pooled human immu (TNF) ligand/receptor superfamily, particularly TNF and US 2017/0044244 A1 Feb. 16, 2017 30

Fas/Fas ligand (FasL) are involved in the pathogenesis of ointment composition of pooled human IgG in a dosage/ MS. Akassoglou et al. 1998 (Am J Pathol. 153(3): 801-813). frequency combination selected from variations 1 to 816 Accordingly, mouse models deficient in TNF can be used to found in Table 1 and Table 2. In a particular embodiment, the study the pathologies of MS. The genotype of transgenic method comprises administering the gel, cream, or ointment TNF knockout mouse models include p55TNFR (p55-/-), composition directly to a nasal epithelium of the Subject. In p75TNFR (p75-/-), and TNF (TNF-/-). a particular embodiment, the method comprises administer (0279 IVIG has proven useful in the treatment of a ing the gel, cream, or ointment composition directly to the number of autoimmune diseases; however its role in the olfactory epithelium of the subject. treatment of MS remains uncertain. IVIG trials in different types of MS patients have produced variable results ranging Parkinson's Disease from reports of monthly IVIG being beneficial to IVIG 0284 Parkinson's disease (PD) is a degenerative disorder administration not slowing disease progression or reversing of the CNS. PD is notably linked to a decrease in motor disease-induced deficits. control. The loss of motor control caused by PD results from 0280. In one aspect, the present invention provides a the death of dopamine-generating cells in the Substantia method for treating multiple Sclerosis in a subject in need nigra, a region of the midbrain. Early in the progression of thereof by delivering a therapeutically effective amount of a the disease, the most common symptoms include shaking, composition comprising pooled human immunoglobulin G rigidity, slowness of movement and difficulty with walking (IgG) to the brain of the subject, wherein delivering the and gait. As the disease progresses, cognitive and behavioral composition to the brain comprises intranasally administer problems arise, with dementia occurring in the advanced ing the composition directly to an epithelium of the nasal stages of the disease. Additional symptoms include sensory, cavity of the Subject. In a specific embodiment, the compo sleep and emotional problems. PD is more common in the sition is administered directly to the olfactory epithelium of elderly, with symptoms most commonly occurring after the the nasal cavity. age of 50. 0281. In one embodiment of the method for treating 0285. There are numerous transgenic mouse models for multiple Sclerosis, the method includes intranasally admin PD. These models include, for example, Park2 (parkin) istering a dry powder composition containing from 0.05 transgenic strains, LRRK2 transgenic strains, and synuclein mg/kg to 50 mg/kg pooled human immunoglobulin to a transgenic strains (Jackson Laboratories, Bar Harbor, Me.). subject in need thereof daily. In other embodiments, the In addition to transgenic models, parkinsonian symptoms methods provided herein for the treatment of multiple scle can also be induced in mice by administering the compounds rosis include intranasally administering a dry powder com MPTP, rotenone, paraquat, or maneb. position of pooled human IgG in a dosage/frequency com 0286. In one aspect, the present invention provides a bination selected from variations 1 to 816 found in Table 1 method for treating Parkinson's disease in a subject in need and Table 2. In a particular embodiment, the method com thereof by delivering a therapeutically effective amount of a prises administering the dry powder composition directly to composition comprising pooled human immunoglobulin G a nasal epithelium of the Subject. In a particular embodi (IgG) to the brain of the subject, wherein delivering the ment, the method comprises administering the dry powder composition to the brain comprises intranasally administer composition directly to the olfactory epithelium of the ing the composition directly to an epithelium of the nasal Subject. cavity of the Subject. In a specific embodiment, the compo 0282. In one embodiment of the method for treating sition is administered directly to the olfactory epithelium of multiple Sclerosis, the method includes intranasally admin the nasal cavity. istering a liquid (e.g., an aqueous) composition containing 0287. In one embodiment of the method for treating from 0.05 mg/kg to 50 mg/kg pooled human immunoglobu Parkinson's disease, the method includes intranasally lin to a subject in need thereof daily. In other embodiments, administering a dry powder composition containing from the methods provided herein for the treatment of multiple 0.05 mg/kg to 50 mg/kg pooled human immunoglobulin to Sclerosis include intranasally administering a liquid (e.g., an a subject in need thereof daily. In other embodiments, the aqueous) composition of pooled human IgG in a dosage/ methods provided herein for the treatment of Parkinson's frequency combination selected from variations 1 to 816 disease include intranasally administering a dry powder found in Table 1 and Table 2. In a particular embodiment, the composition of pooled human IgG in a dosage/frequency method comprises administering the composition drop-wise combination selected from variations 1 to 816 found in Table directly to a nasal epithelium of the Subject. In a particular 1 and Table 2. In a particular embodiment, the method embodiment, the method comprises administering the com comprises administering the dry powder composition position drop-wise directly to the olfactory epithelium of the directly to a nasal epithelium of the Subject. In a particular Subject. In another particular embodiment, the method com embodiment, the method comprises administering the dry prises administering the composition via a spray directly to powder composition directly to the olfactory epithelium of a nasal epithelium of the Subject. In a particular embodi the subject. ment, the method comprises administering the composition 0288. In one embodiment of the method for treating via a spray directly to the olfactory epithelium of the subject. Parkinson's disease, the method includes intranasally 0283. In one embodiment of the method for treating administering a liquid (e.g., an aqueous) composition con multiple Sclerosis, the method includes intranasally admin taining from 0.05 mg/kg to 50 mg/kg pooled human immu istering a gel, cream, or ointment composition containing noglobulin to a subject in need thereof daily. In other from 0.05 mg/kg to 50 mg/kg pooled human immunoglobu embodiments, the methods provided herein for the treatment lin to a subject in need thereof daily. In other embodiments, of Parkinson's disease include intranasally administering a the methods provided herein for the treatment of multiple liquid (e.g., an aqueous) composition of pooled human IgG Sclerosis include intranasally administering a gel, cream, or in a dosage/frequency combination selected from variations US 2017/0044244 A1 Feb. 16, 2017

1 to 816 found in Table 1 and Table 2. In a particular 0300. In one embodiment of the first aspect, at least 50% embodiment, the method comprises administering the com of the pooled human IgG administered to the Subject con position drop-wise directly to a nasal epithelium of the tacts the nasal epithelium of the subject associated with Subject. In a particular embodiment, the method comprises trigeminal nerve endings. administering the composition drop-wise directly to the 0301 In one embodiment of the first aspect, at least 60% olfactory epithelium of the subject. In another particular of the pooled human IgG administered to the Subject con embodiment, the method comprises administering the com tacts the nasal epithelium of the subject associated with position via a spray directly to a nasal epithelium of the trigeminal nerve endings. Subject. In a particular embodiment, the method comprises 0302) In one embodiment of the first aspect, delivering administering the composition via a spray directly to the the composition to the brain comprises intranasally admin olfactory epithelium of the subject. istering the composition to the upper third of the nasal cavity 0289. In one embodiment of the method for treating of the subject. Parkinson's disease, the method includes intranasally 0303. In one embodiment of the first aspect, at least 40% administering a gel, cream, or ointment composition con of the pooled human IgG administered to the Subject con taining from 0.05 mg/kg to 50 mg/kg pooled human immu tacts the upper third of the nasal cavity of the subject. noglobulin to a subject in need thereof daily. In other 0304. In one embodiment of the first aspect, at least 50% embodiments, the methods provided herein for the treatment of the pooled human IgG administered to the Subject con of Parkinson's disease include intranasally administering a tacts the upper third of the nasal cavity of the subject. gel, cream, or ointment composition of pooled human IgG in 0305. In one embodiment of the first aspect, at least 60% a dosage/frequency combination selected from variations 1 of the pooled human IgG administered to the Subject con to 816 found in Table 1 and Table 2. In a particular embodiment, the method comprises administering the gel. tacts the upper third of the nasal cavity of the subject. cream, or ointment composition directly to a nasal epithe 0306 In one embodiment of any of the methods provided lium of the subject. In a particular embodiment, the method above, the CNS disorder is a neurodegenerative disorder of comprises administering the gel, cream, or ointment com the central nervous system. In a specific embodiment, the position directly to the olfactory epithelium of the subject. neurodegenerative disorder of the central nervous system is Alzheimer's disease. In a specific embodiment, the neuro SPECIFIC EMBODIMENTS degenerative disorder of the central nervous system is Par kinson's disease. 0290. In a first aspect, the disclosure provides a method 0307. In one embodiment of any of the methods provided for treating a central nervous system (CNS) disorder in a above, the CNS disorder is a systemic atrophy primarily Subject in need thereof, the method comprising: delivering a affecting the central nervous system. In a specific embodi therapeutically effective amount of a composition compris ment, the systemic atrophy primarily affecting the central ing pooled human immunoglobulin G (IgG) to the brain of nervous system is amyotrophic lateral Sclerosis (ALS). In a the subject, wherein delivering the composition to the brain specific embodiment, the systemic atrophy primarily affect comprises intranasally administering the composition ing the central nervous system is Huntington's disease. directly to a nasal epithelium of the subject. 0308. In one embodiment of any of the methods provided 0291. In one embodiment of the first aspect, at least 40% above, the CNS disorder is an extrapyramidal and move of the pooled human IgG administered to the Subject con ment disorder. tacts the nasal epithelium of the subject. 0309. In one embodiment of any of the methods provided 0292. In one embodiment of the first aspect, at least 50% above, the CNS disorder is a demyelinating disorder of the of the pooled human IgG administered to the Subject con central nervous system. In a specific embodiment, the dem tacts the nasal epithelium of the subject. ylelinating disorder of the central nervous system is multiple 0293. In one embodiment of the first aspect, at least 60% Sclerosis. of the pooled human IgG administered to the Subject con 0310. In one embodiment of any of the methods provided tacts the nasal epithelium of the subject. above, the CNS disorder is an episodic or paroxysmal 0294. In one embodiment of the first aspect, the nasal disorder of the central nervous system. epithelium is the olfactory epithelium of the subject. 0311. In one embodiment of any of the methods provided 0295. In one embodiment of the first aspect, at least 40% above, the CNS disorder is a paralytic syndrome of the of the pooled human IgG administered to the Subject con central nervous system. In a specific embodiment, the CNS tacts the olfactory epithelium of the subject. disorder is a paralytic syndrome of the central nervous 0296. In one embodiment of the first aspect, at least 50% system is cerebral palsy of the pooled human IgG administered to the Subject con 0312. In one embodiment of any of the methods provided tacts the olfactory epithelium of the subject. above, the CNS disorder is a nerve, nerve root, or plexus 0297. In one embodiment of the first aspect, at least 60% disorder of the central nervous system. of the pooled human IgG administered to the Subject con 0313. In one embodiment of any of the methods provided tacts the olfactory epithelium of the subject. above, the CNS disorder is an organic mental disorder. 0298. In one embodiment of the first aspect, the nasal 0314. In one embodiment of any of the methods provided epithelium is a nasal epithelium of the Subject associated above, the CNS disorder is a mental or behavioral disorder with trigeminal nerve endings. caused by psychoactive Substance use. 0299. In one embodiment of the first aspect, at least 40% 0315. In one embodiment of any of the methods provided of the pooled human IgG administered to the Subject con above, the CNS disorder is a schizophrenia, schizotypal, or tacts the nasal epithelium of the subject associated with delusional disorder. In a specific embodiment, the schizo trigeminal nerve endings. phrenia, Schizotypal, or delusional disorder is schizophrenia. US 2017/0044244 A1 Feb. 16, 2017 32

0316. In one embodiment of any of the methods provided to 6.0. In another specific embodiment, the pH of the above, the CNS disorder is a mood (affective) disorder. In a composition is from 6.0 to 7.5. specific embodiment, the mood (affective) disorder is bipo 0329. In one embodiment of any of the methods provided lar disorder. above, the method includes intranasally administering to the 0317. In one embodiment of any of the methods provided subject a dose of from 0.08 mg to 100 mg pooled human IgG above, the CNS disorder is a neurotic, stress-related, or per kg body weight of the Subject (mg IgG/kg). Somatoform disorder. 0330. In one embodiment of any of the methods provided 0318. In one embodiment of any of the methods provided above, the method includes intranasally administering to the above, the CNS disorder is a behavioral syndrome. Subject a dose of from 0.2 mg to 40 mg pooled human IgG 0319. In one embodiment of any of the methods provided per kg body weight of the Subject (mg IgG/kg). above, the CNS disorder is an adult personality or behavior 0331. In one embodiment of any of the methods provided disorder. above, the method includes intranasally administering to the 0320 In one embodiment of any of the methods provided Subject a dose of from 0.5 mg to 20 mg pooled human IgG above, the CNS disorder is a psychological development per kg body weight of the Subject (mg IgG/kg). disorder. 0332. In one embodiment of any of the methods provided 0321. In one embodiment of any of the methods provided above, the method includes intranasally administering to the above, the CNS disorder is a child onset behavioral or Subject a dose of from 0.5 mg to 10 mg pooled human IgG emotional disorder. In a specific embodiment, the child onset per kg body weight of the Subject (mg IgG/kg). behavioral or emotional disorder is Pediatric acute-onset 0333. In one embodiment of any of the methods provided neuropyschiatric syndrome (PANS). In another specific above, the method includes intranasally administering to the embodiment, the child onset behavioral or emotional disor Subject a dose of from 1 mg to 5 mg pooled human IgG per der is Pediatric Autoimmune Neuropsychiatric Disorders kg body weight of the Subject (mg IgG/kg). Associated with Streptococcal infections (PANDAS). 0334. In one embodiment of any of the methods provided 0322. In one embodiment of any of the methods provided above, the method includes intranasally administering to the above, intranasal administration of the composition com subject a fixed dose of from 50 mg to 10 g pooled human prises the use of a non-invasive intranasal delivery device. IgG. 0323. In one embodiment of any of the methods provided 0335. In one embodiment of any of the methods provided above, intranasal administration of the composition com above, the method includes intranasally administering to the prises administration of a liquid drop of the composition subject a fixed dose of from 100 mg to 5 g pooled human directly onto the nasal epithelium. IgG. 0324. In one embodiment of any of the methods provided 0336. In one embodiment of any of the methods provided above, intranasal administration of the composition com above, the method includes intranasally administering to the prises directed administration of an aerosol of the compo subject a fixed dose of from 500 mg to 2.5g pooled human sition to the nasal epithelium. In a specific embodiment, the IgG. aerosol of the composition is a liquid aerosol. In a specific embodiment, the aerosol of the composition is a powder 0337. In one embodiment of any of the methods provided aerosol. above, the method includes intranasally administering to the 0325 In one embodiment of any of the methods provided Subject a dose of pooled human IgG at least twice monthly. above, the composition comprising pooled human IgG does 0338. In one embodiment of any of the methods provided not contain a permeability enhancer. above, the method includes intranasally administering to the 0326 In one embodiment of any of the methods provided Subject a dose of pooled human IgG at least once weekly. above, the composition comprising pooled human IgG con 0339. In one embodiment of any of the methods provided sists essentially of pooled human IgG and an amino acid. In above, the method includes intranasally administering to the a specific embodiment, the amino acid is glycine. In another Subject a dose of pooled human IgG at least twice weekly. specific embodiment, the amino acid is histidine. In another 0340. In one embodiment of any of the methods provided specific embodiment, the amino acid is proline. above, the method includes intranasally administering to the 0327. In one embodiment of any of the methods provided Subject a dose of pooled human IgG at least once daily. above, the composition comprising pooled human IgG is an 0341. In one embodiment of any of the methods provided aqueous composition. In one embodiment, the composition above, the method includes intranasally administering to the comprises: from 10 mg/mL to 250 mg/mL pooled human Subject a dose of pooled human IgG at least twice daily. IgG; and from 50 mM to 500 mM glycine. In a specific 0342. In one embodiment of any of the methods provided embodiment, the pH of the composition is from 4.0 to 7.5. above, the composition comprising pooled human IgG com In another specific embodiment, the pH of the composition prises at least 0.1% anti-amyloid f IgG. is from 4.0 to 6.0. In another specific embodiment, the pH 0343. In one embodiment of any of the methods provided of the composition is from 6.0 to 7.5. above, the method includes administering a second therapy 0328. In one embodiment of any of the methods provided for the CNS disorder to the subject in need thereof. In one above, the composition comprising pooled human IgG is a embodiment, the second therapy for the CNS disorder is a dry powder composition. In one embodiment, the dry pow cholinesterase inhibitor. In a specific embodiment, the cho der composition is prepared from an aqueous Solution com linesterase inhibitor is donepezil. In another specific prising: from 10 mg/mL to 250 mg/mL pooled human IgG; embodiment, the cholinesterase inhibitor is rivastigmine. In and from 50 mM to 500 mM glycine. In a specific embodi another specific embodiment, the cholinesterase inhibitor is ment, the dry powder composition is prepared from an galantamine. In another specific embodiment, the cholinest aqueous solution having a pH of from 4.0 to 7.5. In another erase inhibitor is tacrine. In another embodiment, the second specific embodiment, the pH of the composition is from 4.0 therapy for the CNS disorder is an inhibitor of NMDA-type US 2017/0044244 A1 Feb. 16, 2017

glutamate receptor. In a specific embodiment, the inhibitor TABLE 3-continued of NMDA-type glutamate receptor is memantine. Intranasal administration of IgG to 8 rats to test for intranasal tolerability. Examples # Drops Time to Example 1 Rat Weight (g) Drug? Dose delivered perfusion 4 3.09.00 Liquid protein 10 (a) 6 L/drop 60 min Solution - 100 mg/mL (60 IL total) Tolerability of Intranasal Administration of IgG in 5 342.62 Microsphere - 10 (a) 6 L/drop 60 min Rats 200 mg/mL (60 IL total) 6 355.1 Microsphere - 10 (a) 6 L/drop 60 min 0344) A study was conducted to examine the tolerability 150 mg/mL (60 IL total) of intranasal administration of IgG in rats. The purpose of 7 364.28 Microsphere - 10 (a) 6 L/drop 60 min this study was to determine the tolerability of rats to intra 200 mg/mL (60 IL total) nasal IgG administration at various concentrations and 8 348.93 Microsphere - 28 (a) 6 L/drop 60 min preparations. 150 mg/mL (162 L total) 0345 Experimental Design: 0346 IgG was prepared as a liquid protein solution or as 0350 Results. a microsphere preparation. The liquid IgG protein solution 0351. Three rats received the liquid preparation of intra was prepared in glycine at 200 mg/mL and 100 mg/mL and nasal IgG. One rat received 60 uL at 100 mg/ml and it was had a pH of 5.1-5.3. The IgG microsphere preparation was well tolerated. Two rats received 60 uL at 200 mg/mL. The prepared at 200 mg/mL and 150 mg/mL in PEG. The IgG first rat had some difficulty breathing, most likely due to a preparations were administered to 8 anesthetized, adult male problem with light anesthesia. The second rat had some Sprague Dawley rats. difficulties breathing, but survived. Tracheotomies were not 0347 Prior to anesthesia, each rat was weighed. An necessary. anesthesia cocktail was prepared and full, half, and quarter 0352 Four rats received the microsphere preparation. anesthesia doses were calculated according to the animals Two rats received 60 uL at 150 mg/ml. One rat received 60 weight with a full dose containing 30 mg/kg ketamine, 6 uL at 200 mg/ml. One rat received 162 uL at 150 mg/ml. mg/kg Xylazine, and 1 mg/kg acepromazine. The anesthesia These rats tolerated the highest concentration available at was administered Subcutaneously into the left hind leg, 200 mg/ml very well. above the thigh. Anesthesia was monitored throughout the 0353. The rats tolerated the liquid and microsphere procedures by assessing reflexes using pinching of the hind preparations; however, the rats did tolerate the microsphere paw or tail. If a reflex was present, a half or quarter dose preparation better than the protein preparation. booster was administered as necessary. During drug admin istration, animals received a half dose booster roughly 20-25 Example 2 min after initial dose if needed. 0348 Anesthetized rats were placed on their backs on a Comparison of Liquid, Microsphere, and Fragment heating pad in a metal Surgical tray. The heating pad was Biodistribution at 30 and 90 Minutes connected to a thermostat and was automatically regulated 0354. The purpose of this study was to quantify the to maintain a 37° C. temperature based on continuous amount of intranasally administered IgG that reaches the measurement from a rectal probe. A 2"x2" gauze pad was central nervous system and peripheral tissues in anesthetized rolled tightly into a pillow, taped together, and under the rats. Specifically, the biodistribution of different formula neck to maintain a correct neck position horizontal with the tions and modes of administration were compared. The COunter. different formulations and modes of administration are 0349. A 6 uL drop was loaded into a pipette and wiped described in Table 4. dry with a tissue. A cotton Swab covered in parafilm was used to occlude one naris completely (the flat part of the Swab was pushed gently against the naris to prevent airflow), TABLE 4 while the 6 uL drop was expelled slowly from the pipette Formulations and modes of administration used in biodistribution study. (held at a 45° angle from the rat’s midline), forming a drop on the pipette tip. The drop was lowered onto the open naris *I radiolabeled IgG to be inhaled. The IgG preparations were administered Formulation Mode of Administration intranasally as described in Table 3. Liquid protein Intranasal (biodistribution measured at 30 min formulation post administration) Liquid protein Intravenous biodistribution measured at 30 min TABLE 3 formulation post administration) Liquid protein Intranasal (biodistribution measured at 90 min Intranasal administration of IgG to 8 rats to test for intranasal tolerability. formulation post administration) Microsphere formulation Intranasal (biodistribution measured at 30 min # Drops Time to post administration) Rat Weight (g) Drug? Dose delivered perfusion Microsphere formulation Intranasal (biodistribution measured at 90 min post administration) 1 259.87 Liquid protein 10 (a) 6 L/drop 23 min Microsphere formulation Intranasal (biodistribution measured at 30 min Solution - 200 mg/mL (60 IL total) (low Ci) post administration) 2 272.61 Microsphere - 50 mg/mL 10 (a) 6 L/drop 60 min Antibody fragment (FAb) Intranasal (biodistribution measured at 30 min (60 IL total) 3 309.14 Liquid protein 8 (a) 6 L/drop 60 min post administration) Solution - 200 mg/mL (60 IL total) US 2017/0044244 A1 Feb. 16, 2017 34

0355 Experimental Design: knots. A cotton Swab was used to pump the vein full of 0356 40 male Sprague-Dawley rats were given one of blood. The lateral suture (closest to the knee) was tied into three preparations of 'I radiolabeled IgG. These included a tight knot. A hemostat was attached to the Suture strings of liquid IgG protein solution in glycine at pH 5.1-5.3, IgG in the medial Suture and some tension was added to occlude a microsphere preparation including PEG, or as Fab anti blood flow. body fragments in phosphate buffered saline (PBS). Drug 0361. A 1 mm transverse incision was made in the administration was either intranasal or intravenous. Rats femoral vein and a blunted 25 Gbutterfly needle connected were sacrificed either 30 or 90 min after the onset of delivery to tubing previously filled with 0.9% NaCl and attached to of the IgG preparations for biodistribution studies. a 3-way stopcock was immediately inserted. The medial 0357 For intranasal delivery, the rats were anesthetized Suture was tied down around the needle to secure it in place. and placed on their backs on a heating pad in a metal Surgical To confirm placement within the vein, a small amount of tray. The heating pad was connected to a thermostat and was blood was withdrawn then saline was pushed. Free suture automatically regulated to maintain a 37° C. temperature strings were tied to the butterfly needle securing the cannula based on continuous measurement from a rectal probe. A in place. Muscles were protracted, Sutures securing the limbs 2"x2" gauze pad was rolled tightly into a pillow, taped removed, and the Surgical area was covered with gauze wet together, and under the neck to maintain a correct neck with saline. position horizontal with the counter. A lead impregnated 0362 For the intravenous infusion of 'I IgG, a syringe shield was placed between the Surgical tray and the experi pump was placed in the hood behind the lead shield. Parts menter for protection against radiation. The dose solution, of the pump were covered with parafilm (or Saran wrap) to pipette, pipette tips, and waste receptacle were arranged prevent contamination with radiation. The pump was set for behind the shield for easy access. 4.75 mm diameter and rate of 50 uL/min. The dose solution 0358. A 6 uL drop was loaded into the pipette behind the (48 uL) was mixed with 452 uL of saline (0.9% NaCl, total shield and wiped dry with a tissue. A cotton Swab covered volume 500 uL) in a 1.5 mL microcentrifuge tube. A 1 cc in parafilm was used to occlude one naris completely (the Syringe filled with Saline was attached to the 3-way stopcock flat part of the Swab was pushed gently against the naris to attached to the butterfly needle and placed in the pump. A prevent airflow), while the 6 ul, drop was expelled slowly piece of parafilm was used to secure the Saline Syringe to the from the pipette (held at a 450 angle from the rats midline), stopcock. With the stopcock closed to the rat, the pump was forming a drop on the pipette tip. The drop was lowered onto started to fill the stopcock with saline. the open naris to be inhaled. After two minutes, the alternate 0363 A 1 cc syringe attached to a 27 G or 30 G needle naris was occluded and a 6 LL drop was administered in the was used to collect the drug from the microcentrifuge tube same fashion. A drop was administered as described above and then the Syringe was connected to the 3-way stopcock. every two minutes to alternating nares until a total of8 drops The stopcock was turned so that the flow was open between was delivered (4 to each naris) over 14 min. Delivered time the dose solution and the rat. The tubing was filled with dose of each drop was noted as well as any details regarding the Solution making Sure that no air bubbles are pushed into the animal's respiration or success of the delivery. Three 3 ul. rat and that fluid does not pool near the femoral vein (this aliquots of each dosing Solution were gamma counted to would indicate the needle was not in the vein). The stopcock determine the measured specific activity. was turned so that flow was open to the Saline Syringe and 0359 For intravenous IgG delivery, the rats required the rats. cannulation of the femoral artery. Anesthetized animals were 0364 The time and start volume of the saline syringe was positioned on their backs in Surgical tray on a heating pad noted and the pump was started. The stop volume of the maintained at 37° C. Both hind legs were secured by loosely saline Syringe was also noted at the end of the 14 min tying a Suture around the limbs and weighting them with a infusion. At least 700 u, of saline was infused (50 uL/min hemostat. Small, superficial cuts with blunt scissors were over 14 min). The volume of saline administered was made at the mid inguinal point, making Sure not to cut the slightly more than the volume of the tubing which ensured Superficial blood vessels. Gentle, blunt dissection using that all of the dose solution was administered. cotton Swabs exposed the femoral vein from the great 0365 Two minutes prior to the desired end point time, Saphenous vein to the inguinal ligament. Blunt Scissors were anesthetized animals were laid flat on their backs in a metal used to cut away the skin to get a better view the area. Surgical tray. The heating pad, rectal probe, and neck pillow Overlying muscle was retracted by threading a 4-0 Suture were removed. Tape was used to secure the front limbs to the with a curved needle through the muscle, attaching a curved pan. The back of the pan was elevated slightly to allow blood hemostat to the end of the Suture and weighting it in place. to run away from the animal. The sternum was exposed by Connective tissue Surrounding the femoral vein and artery cutting through the skin. The sternum was clamped with a was carefully removed with blunt dissection (cotton swabs). hemostat and the rib cage was cut open laterally, exposing Connective tissue between the vein and artery was teased the diaphragm. The diaphragm was cut laterally to expose apart using two pairs of forceps carefully using a motion the pleural cavity. running parallel to the blood vessels and being careful not to 0366 Surgical scissors were used to cut up the sides of rupture the vessels. Saline was applied if the area was dry. the ribcage toward the armpits of the animal, creating a 'V' 0360. In an area free of branches, the angled forceps was shaped incision exposing the heart. The hemostat holding inserted underneath the vein, the tip poked through the the sternum was taped above the head to hold the cavity connective tissue, and the forceps slowly opened to pull a 12 open. The heart was stabilized using the blunt forceps while inch 4-0 suture through very carefully. If the vein collapsed, a small cut was made into the left ventricle. A 1 cc-Syringe a cotton Swab was used to gently pump the vein full of with 18 G, 1" blunt needle was inserted into the left ventricle blood. A second Suture was pulled through in a similar and approximately 0.1 mL of blood was removed and placed manner. The medial and lateral sutures were tied into loose into a pre-weighed tube for gamma counting. A second 18 G US 2017/0044244 A1 Feb. 16, 2017

blunt needle attached to an extension set filled with 60 cc of brain tissue in the matrix and placed in respective tubes. The saline was inserted through the left ventricle and into the upper cervical spinal cord was collected. The remaining aorta. A large bulldog clamp was placed just above the heart brain was then bisected along the midline and dissected into on the aorta, securing the blunt needle in place. midbrain, pons, medulla, and cerebellum according to FIG. 0367 The animal was perfused with 60 mL of saline 1G. followed by 360 mL of paraformaldehyde using a syringe 0376 Returning to the head, the ventral side of the neck pump at a rate of 15 mL/min. was cut anteriorly and skin peeled back exposing lymph 0368. Throughout experimental procedures, strict pre nodes, salivary glands, and neck muscles. The Superficial cautions were followed to prevent radioactive contamination nodes, deep cervical nodes, carotid arteries, and thyroid of animal tissues, Surgical tools, and equipment. Geiger gland were dissected and cleared of connective tissue. A counters were placed at each work Station to continuously razor blade was used to bisect the skull along the midline. screen tools, workspace, and staff. Personal protective The olfactory epithelium and respiratory epithelium were equipment including double layered gloves, lab coats, eye collected. protection, masks, and bouflant caps were worn at all times. 0377 For body dissection, bodies were placed on their Lead impregnated shields were used to minimize exposure backs and a longitudinal cut using a scalpel was used to open to radiation. Radioactive monitoring badges were also worn the peritoneal cavity down to the bladder. 3 mm square by staff throughout experimental procedures to quantify samples of liver (superficial right lobe), kidney (left, tip), exposure. renal artery, spleen (tip), lung (right, top lobe), and heart 0369 Immediately after collection, each tissue sample were collected. Approximately 0.1-0.2 mL of urine was was placed into a pre-labeled and pre-weighed gamma tube collected. for later measurement. 0378 Bodies were flipped over onto the stomach and a 0370. For brain dissection, skin and muscle around the Superficial incision was made down the length of the animal neck were cut with a scalpel just above the shoulder blades from shoulders to hips, following the spine. The skin was and a large pair of Scissors used to decapitate the animal, peeled away from the underlying tissue on both sides to cutting dorsal to Ventral to avoid contamination from the expose the shoulder blades. Axillary nodes in the armpits trachea and esophagus. To expose the brain, a midline were dissected and cleared of connective tissue. A piece of incision was made on the dorsal side of the skull, then skin right deltoid muscle was collected (-3 mm). was peeled away, and a straight hemostat was used to break 0379 The muscles overlying the spine were scored with the bone, taking care to leave the dorsal dura attached. a scalpel. To expose the spinal cord, a small hemostat was Dorsal dura was collected. inserted into the spinal column and used to chip away 0371 To remove the brain from the skull, the head was overlying vertebrae and tissues. A Small spatula was used to inverted and a small spatula was used to free it from the loosen the cord from the spinal cavity and forceps used to cavity. The posterior optic nerve and trigeminal nerves were remove it and place into a petri dish. The dura was peeled off cut close to the brain. The brain was then placed into a clean of the cord using forceps. The cord was dissected into lower Petri dish for dissection. cervical, thoracic, and lumbar portions. The top ~2 mm of 0372. From the base of the skull, the ventral dura was lower cervical segment was discarded. collected by Scraping a forceps on the Ventral skull walls. 0380 A 2 cm segment of trachea and esophagus was The pituitary, optic chiasm, and trigeminal nerves were dissected from the body and connective tissues were collected. The anterior portion of the trigeminal nerve con removed. The top 0.5 cm (closest to the decapitation point) sisted of the portion before the visible branch in the skull, of each was discarded. while the remainder containing the trigeminal ganglion was 0381 Pre-weighed gamma tubes containing samples considered as the posterior section. The head was then set were reweighed to determine tissue weight. Tissue samples aside and covered with a kim-wipe for later dissection. from the rats were counted using a COBRA II Auto-Gamma 0373) A microscope was used to help remove vessels Counter using a standard I protocol and a 5 min count from the brain. Using Surgical forceps, microscissors, and a time. Counters were normalized weekly to ensure a counting 30 G needle, the basilar artery and circle of Willis were efficiency at or above 80%. Background counts were sub removed and placed onto pre-weighed paper (paper was tracted. used because of the small weight of this tissue). The needle 0382 Mean and standard error of the nM concentration was used to lift the vessels away from the brain, the forceps of each tissue sample were calculated. Any value outside to grab hold, and the microscissors to make the cuts. This two standard deviations of the mean for each tissue was tissue was weighed immediately upon collection and then considered an outlier and removed from the data set. nM IgG the entire paper was crumpled and placed into the bottom of concentrations were calculated for each tissue using the tube). measured specific activity of dosing solutions, the CPM of 0374 Prior to placing the brain into the coronal matrix, each tissue, and the Volume of each tissue (assuming 1 g 1 the olfactory bulbs were cut off at the natural angle using a mL). razor blade. In the coronal brain matrix, a razor blade was 0383 Results, Intranasal IgG Liquid Preparation Distri inserted at the center of where optic chiasm was before bution at 30 Min End Point. removal to normalize each animal to the same location 0384 Eight rats received IN IgG liquid preparation at an (bregma). Additional blades were placed every 2 mm from average dose of 6.0 mg in 47.4LL containing 69.6 uCi with the first blade, resulting in 6x2 mm slices, 3 rostral to the a 30 min end point. Animals tolerated the IN administration optic chiasm and 3 caudal. well and all survived until the 30 min desired end point. 0375 Blades were removed and tissues were dissected 0385 At the site of IN drug administration, the average from each slice (FIG. 1A-1F). The remaining section of IgG concentrations in the respiratory and olfactory epithelia cortex and hippocampus was dissected from the remaining were 136.213 nM and 442 nM respectively. A rostral to US 2017/0044244 A1 Feb. 16, 2017 36 caudal gradient of 13.1 nM to 6.0 nM. IgG was observed in organs ranged from a low of 1.3 nM in the heart to a high the trigeminal nerve. A similar gradient from the olfactory of 6.1 nM in the spleen and kidney, with urine containing 8.1 bulb to the anterior olfactory nucleus of 4.1 nM to 1.5 nM nM. Concentrations of IgG in the basilar and carotid arteries IgG was observed. The average cortex concentration of IgG were considerably higher than the renal artery (11.7 and 14.1 after IN administration was 1.3 nM. Concentrations of IgG nM versus 4.4 nM). Average concentration of IgG in the in other brain regions ranged from a low of 0.7 nM in the sampled lymph nodes was 4.7 nM. Levels of IgG in tissues striatum to a high of 1.7 nM in the hypothalamus. The measured to assess variability of IN administration and hippocampus was found to contain 0.6 nM IgG. A rostral to breathing difficulty (lung, esophagus, and trachea) were caudal concentration gradient (1.6 nM to 0.7 nM) was consistent across animals. observed within the extra brain material sampled. Similarly, 0387 Results, Intranasal IgG Microsphere Preparation a rostral to caudal concentration gradient (1.2 nM to 0.3 nM) (Low uCi) Distribution at 30 Min EndPoint. was observed in the spinal cord. The average concentration 0388 Four rats received IN IgG microsphere preparation of IgG in the dura of the brain was 15.2 nM compared to a (low LLCi) at an average dose of 7.2 mg in 48.0LL containing spinal cord dura concentration of 2.8 nM. The dura likely 24.7 uCi with a 30 min end point. The raw data from the four also contains some or most of the arachnoid membrane and rats is provided in Table 5. The measured specific activity together comprise two of the three components of the from this dosing solution was much lower than expected meninges. Other tissues sampled from the cavity of the based upon the provided specific activity. Animals tolerated ventral skull (pituitary and optic chiasm) contained 8.2 nM the IN administration well and all survived until the 30 min and 7.4 nM IgG respectively. desired end point. Zero statistically significant outliers and 0386 The blood concentration of IgG at the 30 min end fourteen non-statistically significant outliers were identified point was 13.9 nM. Concentrations of IgG in peripheral out of a total of 211 concentration values. TABLE 5 Biodistribution (nM concentrations) of intranasally administered IgG microsphere preparations (with low uCi) at the 30 min end point with outliers included.

BAX-17 BAX-18 BAX-19 BAX-20 Avg SE Volume Delivered (L) 48.0 48.0 48.0 48.0 48.0 O.OO uCi Delivered 20.9 20.9 28.6 28.6 24.7 2.2 mg Delivered 7.2 7.2 7.2 7.2 7.2 O.OO Olfactory Epithelium 6,806.0 3,931.1 15,573.6 2O3.9 6,628.6 +3,273.6 Respiratory Epithelium 559,241.5 268,256.5 219,595.4 25,412.0 268,126.3 +110,307.7 Anterior Trigeminal 9.7 28.6 11.4 4.6 13.6 5.2 Nerve Posterior Trigeminal 5.4 14.5 6.3 4.1 7.6 +2.4 Nerve Olfactory Bulbs 5.9 3.4 3.7 4.1 4.3 O.6 Anterior Olfactory 1.9 2.4 2.1 1.4 1.9 O.2 Nucleus Frontal Cortex 1.2 1.6 2.O 1.4 1.6 O.2 Parietal Cortex O.8 1.2 1.1 O.S O.9 O.2 Temporal Cortex O.9 : 1.2 O6 O.9 O.2 Occipital Cortex O.O 1.3 0.4 1.6 O.8 +0.4 Extra Cortex 1.6 1.4 1.1 O.9 1.3 O.2 Amygdala 1.6 S.1 1.9 0.7 2.3 O.9 Striatum O.8 16.5 O.6 O6 4.6 +4.0 Septal Nucleus 1.8 4.2 O.6 O.3 1.7 O.9 Hypothalamus 1.8 3.9 2.5 1.1 2.3 O.6 Thalamus O.3 O.9 O.6 0.4 O.S O. Midbrain O.8 1.6 0.7 O6 O.9 O.2 Hippocampus O6 1.3 0.7 0.4 0.7 O.2 Pons O6 1.9 1.2 O.8 1.1 O.3 Medulla 0.7 1.2 1.1 O.8 1.O O. Cerebellum O6 1.2 O.8 O6 O.8 O.2 Extra Slice #1 1.3 2.6 2.4 1.5 2.0 O.3 Extra Slice #2 1.O 1.1 1.2 1.1 1.1 O.05 Extra Slice #3 0.7 1.1 1.O 0.7 O.9 O. Extra Slice #4 0.7 1.2 O.8 O6 O.8 O. Extra Slice #5 O6 1.O O.8 O.S 0.7 O. Extra Slice #6 0.7 1.3 1.O O6 O.9 O.2 Pituitary 7.0 18.1 6.2 3.6 8.7 3.2 Optic Chiasm 14.9 19.0 8.2 8.1 12.5 2.7 Dorsal Dura 12.O 20.7 15.1 2O6 17.1 +2. Ventral Dura 18.5 56.5 16.2 15.2 26.6 10.0 Spinal Dura 2.9 1.O 1.7 3.6 2.3 O.6 Upper Cervical Spinal 1.O 1.2 1.O 1.5 1.2 O. Cord Lower Cervical Spinal 0.4 O.3 0.4 O.9 O.S O. Cord US 2017/0044244 A1 Feb. 16, 2017 37

TABLE 5-continued Biodistribution (nM concentrations) of intranasally administered IgG microsphere preparations (with low uCI) at the 30 min end point with outliers included. BAX-17 BAX-18 BAX-19 BAX-2O Avg SE Thoracic Spinal Cord O.S 0.4 O.6 0.7 O.S O1 Lumbar Spinal Cord O.2 O.2 O.2 O.3 O.2 O.O3 Circle of Willis & Basilar 24.9 29.7 17.7 9.3 20.4 +4.4 Artery Carotid Artery 2O7.3 17.9 14.1 13.1 63.1 48.1 Renal artery (L) 6.1 2.4 4.5 2.4 3.8 O.9 Superficial Nodes (2) 30.8 17.1 6.9 O.8 13.9 6.6 Cervical Nodes (2) 7.7 3.4 9.2 62.7 20.8 +14.0 Axillary Nodes (2) 4.2 2.1 2.2 2.8 2.8 OS Blood Sample 2,889.3 8.0 1,730.1 7.1 1,158.6 705.4 Muscle (R, deltoid) 3.7 2.1 1.2 1.4 2.1 O6 Liver (R, Superficial lobe) 1.2 1.O O.9 O.9 1.O O1 Kidney (L, tip) 13.1 2.5 6.9 3.0 6.4 2.5 Orine 5.4 4.0 9.3 3.0 5.4 +1.4 Spleen (tip) 3.1 1.2 3.0 1.8 2.3 OS Heart 4.4 2.9 0.4 0.7 2.1 O.9 Lung (R, top lobe) 3.4 5.2 2.3 2.6 3.4 O6 Thyroid 28,623.9 102.6 30,320.6 15.7 14,765.7 +8.497.9 Esophagus 3.7 2.5 2.7 4.7 3.4 OS Trachea 2.7 1.5 2.O 4.9 2.8 O.8 Drug Standard CPM 2,316,335 2,316,335 3,256,120 3,256,120 2,786.228 +271,292.6 Drug Standard CPM 2,380,434 2,380.434 3,216.298 3,216.298 2,798,366 +241,293.2 Drug Standard CPM 2,259,775 2,259,775 3,051.466 3,051.466 2,655,621 +228,541.5 * = negative tube weight, so nM could not be calculated

0389 Results, Intranasal IgG Microsphere Preparation Distribution at 30 Min End Point. 0390 Eight rats received IN IgG microsphere preparation at an average dose of 7.2 mg in 48.0LL containing 60.0 uCi with a 30 min end point. The raw data from the eight rats is provided in Table 6. Animals tolerated the IN administration well and all survived until the 30 min desired end point. TABLE 6 Biodistribution (nM concentrations) of intranasally administered IgG microsphere preparations at the 30 min end point with outliers excluded. BAX-21 BAX-22 BAX-23 BAX-25 BAX-26 BAX-28 Avg SE

Volume Delivered 48.0 48.0 48.0 48.0 48.0 48.0 48.0 OOO (LL) uCi Delivered 59.9 56.3 734 60.8 53.1 56.5 6O.O 2.9 mg Delivered 7.2 7.2 7.2 7.2 7.2 7.2 7.2 OOO Olfactory Epithelium X 377.5 629.1 X 97.9 2010 326.4 116.3 Respiratory 23,108.2 20,219.7 33,657.6 87,547.5 183,182.6 101,353.0 74,844.8 +25,792.8 Epithelium Anterior Trigeminal 2.0 1.7 2.1 1.3 O.8 1.2 1.5 O2 Nerve Posterior Trigeminal 2.0 1.2 1.3 O.8 0.7 O.9 1.1 O2 Nerve Olfactory Bulbs 2.7 1.O 1.1 O.6 1.O 0.7 1.2 O3 Anterior Olfactory 0.7 1.2 0.4 0.4 O.3 O.3 O.6 O1 Nucleus Frontal Cortex O.8 O.3 O.8 0.4 0.4 0.4 O.S O1 Parietal Cortex O.3 O.6 O6 O.2 0.4 0.4 0.4 O1 Temporal Cortex O.2 0.4 O.2 1.3 O.2 O.3 O.S O2 Occipital Cortex O.3 O.1 0.4 0.4 2.3 O6 0.7 O3 Extra Cortex O.3 O.3 O.9 O.3 O.2 O.3 0.4 O1 Amygdala O.2 X O.2 0.4 O.3 O.3 O.3 O.04 Striatum O6 1.5 O.3 1.1 X 2.1 1.1 O3 Septal Nucleus 0.4 0.7 O.1 1.1 O.6 O6 O.6 O1 Hypothalamus 0.7 O.S O.3 0.4 O.3 O6 O.S O1 Thalamus O.1 O.1 O.2 O.1 O.1 O.1 O.1 O.O1 Midbrain O.2 O.2 O.S O.2 O.2 O.2 O.3 O.OS Hippocampus O.2 O.3 O.2 O.2 O.1 O.1 O.2 O.O3 Pons 0.4 O.3 O.S O.3 O.S O.3 0.4 O.04 Medulla O.3 O.2 0.4 O.3 O.2 O.2 O.3 O.04 US 2017/0044244 A1 Feb. 16, 2017 38

TABLE 6-continued Biodistribution (nM concentrations) of intranasally administered IgG microsphere preparations at the 30 min end point with outliers excluded. BAX-21 BAX-22 BAX-23 BAX-25 BAX-26 BAX-28 Avg SE

Cerebellum O.3 1.1 X 1.7 O.2 O.2 0.7 O3 Extra Slice #1 1.1 O.3 0.4 0.4 O.3 0.4 O.S O1 Extra Slice #2 O.6 O.2 O.2 O.2 O.2 2.4 O.6 +0.4 Extra Slice #3 0.4 O.3 O.3 O.2 O.2 O.S O.3 O.04 Extra Slice #4 O.3 O.2 O.2 O.2 O.1 2.8 O.6 +0.4 Extra Slice #5 O.2 O.2 O.3 O.2 O.2 O.2 O.2 O.O2 Extra Slice #6 O.2 O.2 O.2 O.2 O.2 O.2 O.2 O.O1 Pituitary 1.8 1.3 3.2 2.9 1.7 2.6 2.2 O3 Optic Chiasm 2.2 1.4 2.5 2.1 1.4 0.7 1.7 O3 Dorsal Dura 5.4 7.8 S.O 2.4 S.1 1.6 4.6 O.9 Ventral Dura 9.4 3.1 2.9 3.7 3.1 1.7 4.0 +1.1 Spinal Dura O.6 0.4 O.8 O.2 X O6 O.S O1 Upper Cervical O.SO O.36 O.S2 O.28 O.34 O.S3 O42 O.04 Spinal Cord Lower Cervical O.O6 O.14 O.13 O.10 O.12 O.12 O.11 O.O1 Spinal Cord Thoracic Spinal O.O6 O.O3 O.08 O.09 X O.04 O.O6 O.O Cord Lumbar Spinal Cord O.08 O.O7 O.10 O.OS O.O7 O.O3 O.O6 O.O1 Circle of Willis & 11.5 X 15.7 12.4 2.0 5.2 9.3 2.5 Basilar Artery Carotid Artery 4.6 5.4 1.6 1.9 X 1.9 3.1 O.8 Renal artery (L) O.9 0.4 O.S 0.7 O6 O.S O.6 O1 Superficial Nodes O.8 0.7 O.9 X 4.3 O.8 1.5 O.7 (2) Cervical Nodes (2) 1.2 1.9 1.1 O.8 X O.S 1.1 O2 Axillary Nodes (2) 0.4 X O.3 O.S 1.O O.S O.S O1 Blood Sample 156.7 261.5 1.1 1.9 362.3 268.7 175.4 61.1 Muscle (R, deltoid) O.1 O.9 O.3 O.3 0.7 O.2 0.4 O1 Liver (R, Superficial O.O X O.1 O.2 O.3 O.3 O.2 O.OS obe) Kidney (L, tip) O.6 O.3 0.4 1.O 1.O 1.2 O.8 O1 Orine O.6 1.1 O.9 O.9 3.5 2.7 1.6 OS Spleen (tip) O.3 0.4 O6 O.6 0.4 O.9 O.S O1 Heart O.3 0.4 O.3 O.1 O.1 O.3 O.3 O.04 Lung (R, top lobe) O.S 0.4 O.3 2.2 O.2 1.3 O.8 O3 Thyroid 1,697.8 3,275.2 16.1 36.2 X 35.6 101.2.2 651.5 Esophagus O.6 0.4 O.1 0.7 1.3 0.4 O.6 O2 Trachea O.S 1.O O.3 O.6 O.8 O6 O.6 O1 Drug Standard 6,936,801 6,170,223 8,071,624 7,024,714 6,006,357 6,587,524 6,799,540.2 +303,198.0 CPM Drug Standard 6,854,563 6,687,656 8,239,126 6,958,531 6,134,932 6,360,075 6,872,480.3 +300,895.5 CPM Drug Standard 6,894,326 6,596,846 9,035,030 7,046,819 6,205,338 6,576,363 7,059,120.2 +412,602.5 CPM X = outlier removed from analysis

0391 At the site of IN drug administration, the average organs ranged from a low of 0.2 nM in the liver to a high of IgG concentrations in the respiratory and olfactory epithelia 0.8 nM in the kidney, with urine containing 1.6 nM. Con were 74,844.8 nM and 326 nM respectively. A rostral to centrations of IgG in the basilar and carotid arteries were caudal gradient of 1.5 nM to 1.1 nM. IgG was observed in the trigeminal nerve. A similar gradient from the olfactory bulb considerable greater than the concentration in the renal to the anterior olfactory nucleus of 1.2 nM to 0.6 nM IgG artery (9.3 and 3.1 nM versus 0.6 nM). Average concentra was observed. The average cortex concentration of IgG after tion of IgG in the sampled lymph nodes was 1.0 nM. Levels IN administration was 0.5 nM. Concentrations of IgG in of IgG in tissues measured to assess variability of IN other brain regions ranged from a low of 0.1 nM in the administration and breathing difficulty (lung, esophagus, thalamus to a high of 1.1 nM in the striatum. The hippocam and trachea) were consistent across animals. IgG levels in pus was found to contain 0.2 nM IgG. The average concen the thyroid varied greatly ranging from 16.1 nM to 3,275 tration of IgG in the extra brain material sampled was 0.4 nM, even after the removal of outliers. nM. similar to the average cortex concentration, and a 0393 Results, Intranasal IgG Fragment Preparation Dis concentration gradient was not observed. A rostral to caudal tribution at 30 Min End Point. concentration gradient (0.42 nM to 0.06 nM) was observed in the spinal cord. The average concentration of IgG in the 0394 Four rats received an IN IgG Fab antibody frag dura of the brain was 4.3 nM compared to a spinal cord dura ment preparation at an average dose of approximately 3.3 concentration of 0.6 nM. Other tissues sampled from the mg in 48.2 LL containing 76.4 uCi. The raw data from the Ventral skull, the pituitary and optic chiasm, contained 2.2 four rats is provided in Table 7. All four experiments were nM and 1.7 nM IgG respectively. completed with an end point of 30 min, and as expected the 0392 The blood concentration of IgG at the 30 min end animals tolerated the IN administration well and all survived point was 175 nM. Concentrations of IgG in peripheral until the desired end point. US 2017/0044244 A1 Feb. 16, 2017 39

TABLE 7 Biodistribution (nM concentrations) of intranasally administered IgG Fab preparations at the 30 min end point with outliers excluded. BAX-41 BAX-42 BAX-43 BAX-44 Avg SE Volume Delivered (L) 48.1 48.1 48.2 48.2 48.2 O.O uCi Delivered 76.7 76.7 76.O 76.O 76.4 O.2 mg Delivered 3.3 3.3 3.3 3.3 3.3 O.O Olfactory Epithelium 232.4 435.2 271.2 X 312.9 62.1 Respiratory Epithelium 93,166.9 138,501.7 59,830.3 140,806.9 108076.4 +19.465.7 Anterior Trigeminal 72.8 101.4 1415 73.3 97.2 16.2 Nerve Posterior Trigeminal 32.4 34.2 33.9 19.8 30.1 3.4 Nerve Olfactory Bulbs 54.O 26.3 45.2 23.7 37.3 7.3 Anterior Olfactory 20.4 14.1 2SO 15.9 18.8 +2.4 Nucleus Frontal Cortex 2O.O 11.9 21.5 X 17.8 3.0 Parietal Cortex 7.0 5.8 1.6 6.7 7.8 1.3 Temporal Cortex S.6 4.3 9.5 4.3 5.9 +1.2 Occipital Cortex 9.3 S.6 7.1 10.0 8.O 1.O Extra Cortex 8.9 7.0 8.5 4.2 7.2 +1.1 Amygdala 10.3 14.3 5.2 6.9 11.7 1.9 Striatum S.O S.O 8.6 3.8 S.6 1.O Septal Nucleus 8.4 6.8 O.8 S.1 7.8 +1.2 Hypothalamus 18.0 18.1 22.7 6.3 16.3 3.5 Thalamus S.1 8.2 9.8 2.9 6.5 1.5 Midbrain 8.9 10.3 1.O 4.0 8.6 +1.6 Hippocampus 6.1 7.4 7.2 2.6 5.8 +1.1 Pons 11.O 12.4 2.4 4.9 10.2 1.8 Medulla 11.O 1O.S 1.3 S.O 9.4 1.5 Cerebellum 9.2 5.5 6.1 8.3 7.3 0.9 Extra Slice #1 27.6 16.8 31.2 32.5 27.0 3.6 Extra Slice #2 12.5 9.8 6.O X 12.8 1.8 Extra Slice #3 8.5 8.1 1.5 13.9 1O.S +1.4 Extra Slice #4 7.4 6.5 9.2 6.2 7.3 O.7 Extra Slice #5 6.8 X 8.1 14.3 9.7 2.3 Extra Slice #6 6.O 5.5 7.4 4.1 5.7 O.7 Pituitary 41.6 44.2 SO.6 34.O 42.6 3.4 Optic Chiasm 31.8 21.8 36.4 12.4 25.6 54 Dorsal Dura 138.2 115.3 129.9 101S 121.2 8.1 Ventral Dura 1231 109.7 106.0 81.1 1OS.O 8.8 Spinal Dura 3.4 8.1 2.7 4.5 4.7 +1.2 Upper Cervical Spinal 2O.S 13.7 16.7 7.9 14.7 2.7 Cord Lower Cervical Spinal 1.O 0.7 O.9 1.3 1.O 0.1 Cord Thoracic Spinal Cord O.9 0.7 O.8 1.3 O.9 0.1 Lumbar Spinal Cord 0.7 O.6 O6 O.8 0.7 0.1 Circle of Willis & 64.O 846 69.8 44.4 65.7 8.3 Basilar Artery Carotid Artery X 36.0 35.5 42.9 38.1 +2.4 Renal artery (L) 9.9 14.8 4.0 5.9 8.6 +2.4 Superficial Nodes (2) 9.0 9.4 5.5 6.7 7.6 0.9 Cervical Nodes (2) 19.5 X 23.8 32.O 25.1 3.7 Axillary Nodes (2) 3.2 6.2 3.6 4.1 4.3 O.7 Blood Sample 31.2 38.4 28.9 33.2 32.9 2.0 Muscle (R, deltoid) 2.87 5.05 2.26 2.18 3.1 O.7 Liver (R, Superficial 3.8 3.3 4.0 2.4 3.4 0.3 obe) Kidney (L, tip) 11.1 21.5 4.0 13.1 12.4 3.6 Orine 10.6 10.3 19.9 9.0 12.4 2.5 Spleen (tip) 9.7 12.9 3.4 9.0 8.7 2.0 Heart O.8 3.0 4.5 1.5 2.5 0.8 Lung (R, top lobe) 3.5 9.1 6.7 4.4 5.9 +1.2 Thyroid 228.2 411.7 230.1 273.2 285.8 43.2 Esophagus 4.1 6.4 X 5.8 5.4 O.7 Trachea S.6 8.7 11.3 4.8 7.6 1.5 Drug Standard CPM 7,158,905 7,158,905 6,994.454 6,994.454 7076679.3 +47,472.8 Drug Standard CPM 6,974,631 6,974,631 7.215,418 7.215,418 7095.024.0 +69,509.2 Drug Standard CPM 7,280,104 7,280,104 7,020,805 7,020,805 7150454.3 +74,853.3 X = outlier removed from analysis US 2017/0044244 A1 Feb. 16, 2017 40

0395. At the site of IN drug administration, the average 0396 The blood concentration of IgG Fab at the 30 min IgG Fab concentrations in the respiratory and olfactory end point was 32.9 nM. Concentrations of IgG Fab in epithelia were 108,076 nM and 313 nM respectively. A peripheral organs ranged from a low of 2.5 nM in the heart rostral to caudal gradient of 97.2 nM to 30.1 nM. IgG Fab to a high of 12.4 nM in the kidney and urine, with the spleen was observed in the trigeminal nerve. A similar gradient from the olfactory bulb to the anterior olfactory nucleus of containing 8.7 nM. Concentrations of IgG Fab in the basilar 37.3 nM to 18.8 nM. IgG Fab was observed. The average and carotid arteries were considerably higher than the renal cortex concentration of IgG Fab after IN administration was artery (65.7 and 38.1 nM versus 8.6 nM). 9.3 nM. Concentrations of IgG in other brain regions ranged 0397 Results, Comparison of 30 Min and 90 Min End from a low of 5.6 nM in the striatum to a high of 16.3 nM points. in the hypothalamus. The hippocampus was found to contain 5.8 nM IgG Fab. A rostral to caudal concentration gradient 0398 Concentrations of IgG in brain tissues were gen (27.0 nM to 5.7 nM) was observed within the extra brain erally similar or slightly higher with the extended 90 min material sampled. Similarly, a rostral to caudal concentration end point as compared to the 30 min end point for the IgG gradient (14.7 nM to 0.7 nM) was observed in the spinal liquid preparation. There was more variability in the IgG cord. The average concentration of IgG Fab in the dura of microsphere preparation, with Some tissues containing much the brain was 113.1 nM compared to a spinal cord dura more (thalamus, midbrain) and some tissues containing concentration of 4.7 nM. Other tissues sampled from the much less (striatum, occipital cortex) at the 90 min vs. the cavity of the ventral skull (pituitary and optic chiasm) 30 min end points. Summaries of the IgG concentrations in contained 42.6 nM and 25.6 nM IgG Fab respectively. tissues are provided in Table 8 and Table 9. TABLE 8 Summary of tissue concentrations (nM + SE) of IN, IV, and Fab IgG at 30 min and 90 min endpoints with outliers removed. Treatment IgG Protein (mean nM + SE) Route

Intravenous Intranasal Time Point

30 min 90 min Sample Size

n = 7 n = 8 n = 6 Volume Delivered (IL) 47.7 O2 47.4 O2 47.6 0.1 uCi Delivered 69.5 O.3 69.6 O.3 7O.O. O.O1 mg Delivered 6.O. O.O3 6.O. O.O2 7.4 O.OO Olfactory Epithelium 43.0 - 3.7 441 - 185 3SS 71 Respiratory 41.1 + 4.3 136,213 + 27,325 163,627 + 16,376 Epithelium Anterior Trigeminal 10.5 - 1.0 13.1 2.6 19.3 2.8 Nerve Posterior Trigeminal 6.3 1.O 6.O. 11 8.4 1.7 Nerve Olfactory Bulbs 3.4 O.S 4.1 - 0.9 9.9 1.6 Anterior Olfactory 1.9 O.3 1.5 + 0.2 2.5 O.3 Nucleus Frontal Cortex 2.9 - O.S 1.4 + O. 3.8 - 0.6 Parietal Cortex 3.3 O.7 O.9 O. 5 - 0.1 Temporal Cortex 2.90.7 1.1 + O. 4 - 0.2 Occipital Cortex 2.3 O2 1.8 O.3 2.5 O.2 Extra Cortex 1.8 O.3 1.O. O. 9 O.2 Amygdala 1.9 O.1 1.4 + 0.2 6 - 0.2 Striatum 1.8 O2 O.7 O. O.9 O.1 Septal Nucleus 1.8 O.1 O.9 O. 1 - 0.1 Hypothalamus 2.0 O2 1.7 O.3 9 O.2 Thalamus 1.7 O.3 O4 O.O3 O.6 O.04 Midbrain 1.8 O.3 O.7 O. 3 - 0.1 Hippocampus 1.1 + 0.1 O.6 O. O. O.1 Pons 1.7 O2 O.9 O. 6 - 0.2 Medulla 1.8 O.3 O.9 O. 6 - 0.2 Cerebellum 1.9 O.3 O.8 O. 7 O.2 Extra Slice #1 2.0 O2 1.6 O.3 3.3 O.4 Extra Slice #2 2.1 O.3 1.O. O. 9 O.2 Extra Slice #3 22 O.3 O.8 O. 6 - 0.2 Extra Slice #4 2.4 + 0.4 O.7 O. 2 - 0.1 Extra Slice #5 26 - 0.6 O.7 O. 2 - 0.1 Extra Slice #6 26 - O.S O.9 O. 3 - 0.1 Pituitary 10.1 - 0.8 8.2 - 18 8.4 + 1.1 Optic Chiasm 5.1 O.7 7.4 1.7 8.O. O.7 Dorsal Dura 27.6 3.0 15.3 2.6 31.1 - 4.0 US 2017/0044244 A1 Feb. 16, 2017 41

TABLE 8-continued Summary of tissue concentrations (nM + SE) of IN, IV, and Fab IgG at 30 min and 90 min endpoints with outliers removed. Ventral Dura 23.5 - 31 15.0 - 2.5 32.3 - 4.4 Spinal Dura 47.2 3.0 2.8 O.3 3.3 O.7 Upper Cervical Spinal 2.0 O2 1.2 - 0.1 2.0 O.3 Cord Lower Cervical Spinal 2.6 O.3 O.6 0.1 O.6 O.1 Cord Thoracic Spinal Cord 1.6 O2 O.4 0.1 O.S. O.1 Lumbar Spinal Cord 2.1 O.3 O3 + O.04 O4 O.1 Circle of Willis & 18.1 2.8 11.7 - 2.5 14.8 + 1.1 Basilar Artery Carotid Artery 33.23.3 14.1 + 2.0 16.1 2.3 Renal artery (L) 111.2 - 10.1 4.4 + 1.0 114 - 3.3 Superficial Nodes (2) 25.3 2.9 4.8 + 0.4 10.4 2.2 Cervical Nodes (2) 62.6 9.2 5.6 O.7 6.9 O.8 Axillary Nodes (2) 42.8 12.8 3.7 O.S 6.0 - 0.6 Blood Sample 1,361 + 42.5 13.9 O.9 19.7 1.4 Muscle (R, deltoid) 19.1 - 3.8 2.7 - O.S 2.90.7 Liver (R, Superficial 135 23.7 1.7 O2 2.6 O.4 obe) Kidney (L, tip) 3SS 30.8 6.1 O.8 8.5 - 1.6 Orine 92.6 26.0 8.1 + 1.4 17.5 - 22 Spleen (tip) 228 17.5 6.1 - 1.0 6.8 0.4 Heart 63.2 - 11.7 1.3 O2 2.7 O.6 Lung (R, top lobe) 261 - S13 2.90.4 4.5 - 0.7 Thyroid 534 65.O 148 - 12.8 620 i 30.8 Esophagus 28.1 - 39 43 O.6 7.7 1.3 Trachea 28.2 6.2 3.9 0.6 6.6 t 1.4 Drug Standard CPM 7,448,243 + 128,562 7,630,853 + 169,309 7,166,204 + 76,377 Drug Standard CPM 7,089,796 + 272,234 7,470,182 + 171,868 7,200.437 + 154,753 Drug Standard CPM 7,390,784 + 351,624 7,689,073 + 214,590 7,022,761 + 10,481

Treatment IgG Microspheres (mean IgG FAB nM + SE) (mean nM + SE) Route

Intranasal Intranasal Time Point

30 min 90 min 30 min Sample Size

n = 6 n = 5 n = 4 Volume Delivered (L) 48.O. O.OO 48.O. O.OO 48.2 O.O uCi Delivered 60.0 - 29 59.7 2.0 76.4 O2 mg Delivered 7.2 OOO 7.2 OOO 3.3 O.O Olfactory Epithelium 326 116 3,192 + 1,625 312.9 62.1 Respiratory 74,845 + 25,793 124,509 + 20,723 108076.4 + 19.465.7 Epithelium Anterior Trigeminal 1.5 + 0.2 8.0 - 1.3 97.2 16.2 Nerve Posterior Trigeminal 30.1 - 3.4 Nerve Olfactory Bulbs 37.3 7.3 Anterior Olfactory 18.8 24 Nucleus Frontal Cortex 17.8 3.0 Parietal Cortex 7.8 1.3 Temporal Cortex 5.9 1.2 Occipital Cortex 8.0 - 1.0 Extra Cortex 7.2 - 1.1 Amygdala 11.7 - 1.9 Striatum 5.6 1.0 Septal Nucleus 7.8 1.2 Hypothalamus 16.3 3.5 Thalamus 6.5 - 1.5 Midbrain 8.6 1.6 Hippocampus 5.8 1.1 Pons 10.2 - 18 Medulla 94 - 1.5 Cerebellum 7.3 O.9 Extra Slice #1 27.036 US 2017/0044244 A1 Feb. 16, 2017 42

TABLE 8-continued Summary of tissue concentrations (nM + SE) of IN, IV, and Fab IgG at 30 min and 90 min endpoints with outliers removed. Extra Slice #2 O.6 0.4 O.S. O.1 12.8 1.8 Extra Slice #3 O.3 0.04 O.4 0.04 10.5 - 1.4 Extra Slice #4 O.6 0.4 O3 + O.O3 7.3 O.7 Extra Slice #5 O.2 O.O2 O.3 0.04 9.7 2.3 Extra Slice #6 O.2 OO1 O3 + O.OS 5.7 O.7 Pituitary 2.2 O.3 2.8 OS 42.6 3.4 Optic Chiasm 17 O.3 1.9 O.9 25.654 Dorsal Dura 4.6 O.9 5.8 1.8 121.2 - 8.1 Ventral Dura 4.0 + 1.1 114 - 3.8 105.0 - 8.8 Spinal Dura O.S. O.1 O.7 0.1 4.7 1.2 Upper Cervical Spinal O.4 0.04 O.6 O2 14.7 - 2.7 Cord Lower Cervical Spinal O.1 OO1 O3 + 0.1 1.O. O.1 Cord Thoracic Spinal Cord O.1 OO1 O.2 O.OS O.9 O.1 Lumbar Spinal Cord O.1 OO1 O.2 O.O2 O.7 O.1 Circle of Willis & 9.3 2.5 5.8 11 65.7 8.3 Basilar Artery Carotid Artery 3.10.8 6.3 O.4 38.1 24 Renal artery (L) O.6 0.1 3.7 1.5 8.6 24 Superficial Nodes (2) 1.5 + 0.7 2.4 + 0.1 7.6 O.9 Cervical Nodes (2) 1.1 O2 2.6 O.O3 25.1 3.7 Axillary Nodes (2) O.S. O.1 2.6 0.6 4.3 O.7 Blood Sample 175 61 223 84.2 32.9 - 2.0 Muscle (R, deltoid) O.4 0.1 O.9 O.3 3.1 O.7 Liver (R, Superficial 3.4 O.3 obe) Kidney (L, tip) O.8 0.1 2.6 0.6 124 3.6 Orine 1.6 OS 6.3 1.7 124 2.5 Spleen (tip) O.S. O.1 2.0 - O.S 8.7 2.O Heart 0.3 it 0.04 0.6 - 0.1 2.5 O.8 Lung (R, top lobe) O.8 O.3 1.2 + 0.4 5.9 1.2 Thyroid 1,012 + 652 216 SO 285.8 43.2 Esophagus O.6 O2 4.7 - 1.3 54 - O.7 Trachea O.6 0.1 2.0 - 0.6 7.6 1.5 Drug Standard CPM 6,799,540 + 303,198 6,861,351 + 210,321 7076679.3 + 47.472.8 Drug Standard CPM 6,872,480 + 300,896 6,758,588 + 176,717 7095.024.0 + 69,509.2 Drug Standard CPM 7,059,120 + 412,602 7,027,097 + 316,344 7150454.3 + 74,853.3

TABLE 9 Summary of tissue concentrations of IgG normalized to a 6 Ing dose. Treatment IgG Microspheres IgG FAB IgG Protein (mean nM) (mean nM) (mean nM) Route

Intravenous Intranasal Intranasal Intranasal Time Point

30 min 90 min 30 min 90 min 30 min Sample Size

n = 7 l = 8 n = 6 l = 6 n = 5 l Volume Delivered (L) 47.7 47.4 47.6 48.0 48.0 48.2 uCi Delivered 69.5 69.6 7O.O 6O.O 59.7 76.4 mg Delivered 6.O 6.0 7.4 7.2 7.2 3.3 Olfactory Epithelium 43.0 441 288 272 2,660 569.O Respiratory Epithelium 41.1 136 213 132,671 62, 371 103,758 196 SO2.6 Anterior Trigeminal Nerve 1O.S 13.1 15.6 1.3 6.7 176.8 Posterior Trigeminal Nerve 6.3 6.0 6.8 O.9 2.6 54.7 Olfactory Bulbs 3.4 4.1 8.0 1.O 1.2 67.8 Anterior Olfactory Nucleus 1.9 1.5 2.1 O.S O.S 34.3 Frontal Cortex 2.9 1.4 3.1 0.4 O.S 32.3 Parietal Cortex 3.3 O.9 1.3 O.3 O.3 14.1 Temporal Cortex 2.9 1.1 1.1 0.4 0.4 10.8 Occipital Cortex 2.3 1.8 2.0 O6 O.3 14.5 Extra Cortex 1.8 1.O 1.6 O.3 0.4 13.0 US 2017/0044244 A1 Feb. 16, 2017 43

TABLE 9-continued Summary of tissue concentrations of IgG normalized to a 6 Ing dose. Treatment IgG Microspheres IgG FAB IgG Protein (mean nM) (mean nM) (mean nM) Route

Intravenous Intranasal Intranasal Intranasal Time Point

30 min 90 min 30 min 90 min 30 min Sample Size

n = 7 n = 8 : 6 n = 6 n = 5 n = 4

Amygdala 1.9 1.4 O.2 O.3 21.2 Striatum 1.8 0.7 O.9 O.S 10.2 Septal Nucleus 1.8 O.9 O.S O.S 14.2 Hypothalamus 2.0 1.7 0.4 O.S 29.6 Thalamus 1.7 0.4 O.1 O.3 11.8 Midbrain 1.8 0.7 O.2 0.4 15.6 Hippocampus 1.1 O6 O.2 0.4 10.6 Pons 1.7 O.9 O.3 0.4 18.5 Medulla 1.8 O.9 O.2 O.3 17.1 Cerebellum 1.9 O.8 O6 0.4 13.2 Extra Slice #1 2.0 1.6 0.4 O.6 49.1 Extra Slice #2 2.1 1.O O.S 0.4 23.2 Extra Slice #3 2.2 O.8 O.2 O.3 19.1 Extra Slice #4 2.4 0.7 O.S O.2 13.3 Extra Slice #5 2.6 0.7 O.2 O.3 17.7 Extra Slice #6 2.6 O.9 O.2 O.3 10.4 Pituitary 10.1 8.2 1.9 2.3 77.5 Optic Chiasm S.1 7.4 6.5 1.4 1.6 46.5 Dorsal Dura 27.6 15.3 25.2 3.8 4.9 220.4 Ventral Dura 23.5 1S.O 26.2 3.3 9.5 190.8 Spinal Dura 47.2 2.8 2.7 0.4 O.6 8.5 Upper Cervical Spinal Cord 2.0 1.2 1.6 0.4 O.S 26.7 Lower Cervical Spinal Cord 2.6 O6 O.S O.1 O.3 1.8 Thoracic Spinal Cord 1.6 0.4 0.4 O.1 O.2 1.7 Lumbar Spinal Cord 2.1 O.3 0.4 O.1 O.1 1.2 Circle of Willis & Basilar Artery 18.1 11.7 12.0 7.8 4.8 119.4 Carotid Artery 33.2 14.1 13.1 2.6 5.3 69.3 Renal artery (L) 111.2 4.4 9.2 O.S 3.1 15.7 Superficial Nodes (2) 25.3 4.8 8.5 1.2 2.O 13.9 Cervical Nodes (2) 62.6 S.6 S.6 O.9 2.2 45.6 Axillary Nodes (2) 42.8 3.7 4.9 0.4 2.2 7.8 Blood Sample 1,361 13.9 16.0 146 186 59.8 Muscle (R, deltoid) 19.1 2.7 2.3 0.4 O.8 S.6 Liver (R, Superficial lobe) 135 1.7 2.1 O.2 0.7 6.1 Kidney (L, tip) 355 6.1 6.9 O6 2.1 22.6 Orine 92.6 8.1 14.2 1.3 5.2 22.6 Spleen (tip) 228 6.1 5.5 0.4 1.7 15.9 Heart 63.2 1.3 2.2 O.2 O.S 4.5 Lung (R, top lobe) 261 2.9 3.6 0.7 1.O 10.8 Thyroid 534 148 5O2 843 18O 519.6 Esophagus 28.1 4.3 6.2 O.S 3.9 9.9 Trachea 28.2 3.9 5.4 O.S 1.6 13.8

0399 Results, Intranasal IgG Liquid Preparation Distri a 90 min end point. Animals tolerated the IN administration bution at 90 Min End Point. well and all survived until the 90 min desired end point. The nanomolar IgG concentrations in tissues for IN IgG liquid 0400 Six rats received IN IgG liquid preparation at an preparation administrations taken at the 90 min end point are average dose of 7.4 mg in 47.6 uL containing 70.0 uCi with presented in Table 10. TABLE 10 Tissue concentrations (nM) of IgG after intranasal IgG liquid preparation administration at the 90 min end point with outliers excluded. BAX-24 BAX-33 BAX-34 BAX-3S BAX-36 BAX-40 Avg SE

Volume 47.8 47.4 47.4 47.5 47.5 47.8 47.6 O1 Delivered (L) US 2017/0044244 A1 Feb. 16, 2017 44

TABLE 10-continued Tissue concentrations (nM) of IgG after intranasal IgG liquid preparation administration at the 90 min end point with Outliers excluded. BAX-24 BAX-33 BAX-34 BAX-3S BAX-36 BAX-40 Avg SE uCi Delivered 70.O 7O.O 7O.O 70.O 7O.O 70.O 7O.O O.O1 mg Delivered 7.4 7.4 7.4 7.4 7.4 7.4 7.4 O.OO Olfactory 669.3 389.3 196.5 2O3.8 307.7 365.8 355.4 70.8 Epithelium Respiratory 205,721.0 194,945.7 189,.621.1 139,524.3 150,482.2 101,469.9 163,627.4 +16,376.5 Epithelium Anterior 16.8 3O.O 1S.O 10.6 20.1 23.1 19.3 2.8 Trigeminal Nerve Posterior X 13.4 5.2 6.7 S.6 11.3 8.4 1.7 Trigeminal Nerve Olfactory 15.5 9.1 10.1 1O.S 3.6 10.8 9.9 +1.6 Bulbs Anterior 3.0 3.4 2.3 2.7 1.5 2.4 2.5 O.3 Olfactory Nucleus Frontal 3.3 5.9 2.4 S.O 2.7 3.3 3.8 O.6 Cortex Parietal 4 1.6 3 2.1 4 X 5 O.1 Cortex Temporal .1 O.9 7 1.4 .2 2.0 .4 O.2 Cortex Occipital 2.1 3.43 8 2.6 2.4 2.8 2.5 O.2 Cortex Extra Cortex .6 .6 .6 2.6 9 2.5 9 O.2 Amygdala 7 .6 3 1.2 .1 2.6 .6 O.2 Striatum O.9 0.4 O6 1.O .2 1.O O.9 O.1 Septal .4 .1 .1 O.9 .4 O.9 .1 O.1 Nucleus Hypothalamus 2.5 8 .6 2.3 3 2.2 9 O.2 Thalamus O.6 O6 O6 O.S O6 O.78 O6 O.04 Midbrain 4 .1 .4 1.O .2 1.8 3 O.1 Hippocampus O O.9 .1 0.7 O.8 1.2 O O.1 Pons .9 .2 .1 1.9 .2 2.2 .6 O.2 Medulla 7 O .1 1.9 .5 2.4 .6 O.2 Cerebellum .4 O .6 2.1 .4 2.5 7 O.2 Extra Slice #1 3.6 4.2 2.5 4.1 .6 3.7 3.3 +0.4 Extra Slice #2 X 2.4 .4 2.1 .4 2.3 9 O.2 Extra Slice #3 .4 9 .1 1.6 .1 2.3 .6 O.2 Extra Slice #4 .2 S6 O.8 1.2 .1 1.2 .2 O.1 Extra Slice #5 2 3 O.9 1.1 .1 1.43 .2 O.1 Extra Slice #6 5 .2 O 1.1 O 1.9 3 O.1 Pituitary 12.7 5.5 S.6 8.1 8.4 10.4 8.4 +1.1 Optic Chiasm 7.2 8.4 7.1 9.9 5.7 9.6 8.0 O.7 Dorsal Dura 12.3 33.0 37.7 37.7 29.6 36.5 31.1 +4.0 Ventral Dura 21.6 47.6 41.2 26.5 21.4 35.4 32.3 +4.4 Spinal Dura 2.0 3.2 .4 2.6 4.3 6.4 3.3 O.7 Upper 2.8 1.2 9 1.8 .5 2.8 2.0 O.3 Cervica Spinal Cord LOWe 0.7 O6 0.4 0.7 0.7 X O6 O.1 Cervica Spinal Cord Thoracic O.S 0.4 0.4 O.3 O.S O.9 O.S O.08 Spinal Cord Lumbar 0.4 O.3 O.2 0.4 O.S O.8 0.4 O.08 Spinal Cord Circle of 16.8 11.7 15.1 X 17.7 12.9 14.8 +1.1 Willis & Basilar Artery Carotid 17.4 13.5 12.6 13.0 13.5 26.9 16.1 2.3 Artery Renal artery 22.4 8.4 7.7 14.6 3.7 X 11.4 3.3 (L) Superficial 7.4 2O.S S.O 8.7 9.6 11.5 10.4 +2.2 Nodes (2) Cervical 6.7 8.9 4.4 5.2 6.9 9.1 6.9 O.8 Nodes (2) Axillary 7.2 5.8 4.3 4.5 7.9 6.3 6.O O.6 Nodes (2) US 2017/0044244 A1 Feb. 16, 2017 45

TABLE 10-continued Tissue concentrations (nM) of IgG after intranasal IgG liquid preparation administration at the 90 min end point with Outliers excluded. BAX-24 BAX-33 BAX-34 BAX-3S BAX-36 BAX-40 Avg SE Blood Sample 17.7 23.3 16.1 X 22.5 19.0 19.7 +1.4 Muscle (R, 2.6 3.2 1.O 1.3 5.4 3.6 2.9 O.7 deltoid) Liver (R, 2.8 1.4 3.9 1.1 2.7 3.4 2.6 +0.4 Superficial obe) Kidney (L, tip) 16.O 7.6 8.8 4.7 6.6 7.3 8.5 +1.6 Orine 25.9 10.8 18.7 12.3 18.0 19.3 17.5 +2.2 Spleen (tip) 5.8 7.8 6.8 7.1 5.3 7.7 6.8 +0.4 Heart 2.1 4.8 1.5 X 1.8 3.4 2.7 O6 Lung (R, top 5.4 2.0 4.6 7.3 4.5 3.1 4.5 O.7 obe) Thyroid 543.6 566.7 700.5 X 680.7 606.1 619.5 3O8 Esophagus 13.5 8.7 6.3 7.4 5.5 4.8 7.7 1.3 Trachea 13.5 6.6 S.6 3.4 S.6 S.O 6.6 +1.4 Drug 7,390,846 7,130,719 7,130,719 6,977,049 6,977,049 7,390,846 7166204.3 +76,377.4 Standard CPM Drug 7,285,169 7,575,479 7,575,479 6,740,664 6,740,664 7,285,169 7200437.0 +154,753.0 Standard CPM Drug 6,990,473 7,032,426 7,032,426 7,045,383 7,045,383 6,990.473 7022760.5 +10,480.7 Standard CPM

04.01 X-outlier removed from analysis. At the site of IN 0402. The blood concentration of IgG at the 30 min end drug administration, the average IgG concentrations in the point was 19.7 nM. Concentrations of IgG in peripheral respiratory and olfactory epithelia were 163,627 nM and 355 organs ranged from a low of 2.6 nM in the liver to a high of nM respectively. A rostral to caudal gradient of 19.3 nM to 7.7 nM in the spleen, with urine containing 17.5 nM. Concentrations of IgG in the basilar and carotid arteries 8.4 nM IgG was observed in the trigeminal nerve. A similar were similar to the concentration in the renal artery (14.8 gradient from the olfactory bulb to the anterior olfactory and 16.1 nM versus 11.4 nM). Average concentration of IgG nucleus of 9.9 nM to 2.5 nM IgG was observed. The average in the sampled lymph nodes was 7.8 nM. Levels of IgG in cortex concentration of IgG after IN administration was 2.2 tissues measured to assess variability of IN administration nM. Concentrations of IgG in other brain regions ranged and breathing difficulty (lung, esophagus, and trachea) were from a low of 0.6 nM in the thalamus to a high of 1.9 nM consistent across animals. in the hypothalamus. The hippocampus was found to contain 0403. Results, Intranasal IgG Microsphere Preparation 1.0 nM. IgG. A rostral to caudal concentration gradient (3.3 Distribution at 90 Min End Point. nM to 1.2 nM) was observed within the extra brain material 04.04 Six rats received IN IgG microsphere preparation sampled. Similarly, a rostral to caudal concentration gradient at an average dose of 7.2 mg in 48.0LL containing 59.7 uCi (2.0 nM to 0.4 nM) was observed in the spinal cord. The with a 90 min end point. Animals tolerated the IN admin average concentration of IgG in the dura of the brain was istration well and all survived until the 90 min desired end 31.7 nM compared to a spinal cord dura concentration of 3.3 point. The nanomolar IgG concentrations in tissues for IN nM. Other tissues sampled from the ventral skull, the IgG microsphere preparation administrations taken at the 90 pituitary and optic chiasm, contained 8.4 nM and 8.0 nM min end point in five of the six rats are presented in Table IgG respectively. 11. TABLE 11 Tissue concentrations (nM) of IgG after intranasal IgG microsphere preparation administration at the 90 min end point with outliers excluded.

BAX-31 BAX-32 BAX-37 BAX-38 BAX-39 Avg SE

Volume Delivered (IL) 48.0 48.0 48.0 48.0 48.0 48.0 O.OO uCi Delivered 57.5 52.7 62.8 62.8 62.8 59.7 2.O mg Delivered 7.2 7.2 7.2 7.2 7.2 7.2 O.OO Olfactory Epithelium 293.5 3,632.7 853.2 1898.5 9,281.1 3,191.8 +1,625 Respiratory Epithelium 169,083.6 169,807.3 128,471.3 69.460.0 85,723.8 124,509.2 +20,723 Anterior Trigeminal 11.1 5.9 10.7 7.6 4.5 8.O 1.3 Nerve Posterior Trigeminal 2.6 3.3 3.6 X 3.1 3.1 O.2 Nerve US 2017/0044244 A1 Feb. 16, 2017 46

TABLE 11-continued Tissue concentrations (nM) of IgG after intranasal IgG microsphere preparation administration at the 90 min end point with Outliers excluded. BAX-31 BAX-32 BAX-37 BAX-38 BAX-39 Avg SE Olfactory Bulbs 2.0 1.9 1.2 1.2 1.2 1.5 O.2 Anterior Olfactory 0.7 O.S O.S 0.4 O.8 O6 O. Nucleus Frontal Cortex 0.7 O.8 0.4 O.8 O.6 0.7 O. Parietal Cortex O.3 X O.1 0.4 O.S O.3 O. Temporal Cortex 0.7 0.7 O.3 O.S O.S O.S O. Occipital Cortex O6 0.4 O.1 0.4 O.2 O.3 O. Extra Cortex O6 O.81 0.4 0.4 X O.S O. Amygdala O.2 X O.3 O.49 O.S3 0.4 O. Striatum O.2 1.3 O.2 0.4 O.9 O6 O.2 Septal Nucleus 0.4 1.9 O.1 O.2 X O6 +0.4 Hypothalamus O6 O.8 0.4 0.4 O.9 O6 O. Thalamus O.2 O.6 O.1 O.2 0.4 O.3 O. Midbrain O.3 O.S O.2 X O.8 O.S O. Hippocampus O.3 O.S O.2 O.2 1.O O.S O. Pons O.S 0.7 0.4 O.S O.S O.S O. Medulla O.S 0.4 O.3 0.4 O.S 0.4 O.04 Cerebellum O.S O.8 O.2 0.4 0.7 O.S O. Extra Slice #1 1.O 1.O 0.4 O.8 0.7 O.8 O. Extra Slice #2 0.4 O.SO O.2 0.4 O.96 O.S O. Extra Slice #3 O.3 0.4 O.2 0.4 O.S 0.4 O.04 Extra Slice #4 O.3 X O.2 O.3 O.3 O.3 O.O3 Extra Slice #5 O.3 0.4 X O.S O.3 O.3 O.04 Extra Slice #6 0.4 X O.2 0.4 0.4 O.3 O.OS Pituitary 4.4 2.1 2.9 2.7 1.6 2.8 O.S Optic Chiasm X 3.4 X 1.9 0.4 1.9 O.9 Dorsal Dura X 11.3 3.8 3.7 4.5 5.8 1.8 Ventral Dura 11.8 26.O 7.4 3.9 8.1 11.4 3.8 Spinal Dura O6 O.8 0.7 X X 0.7 O.1 Upper Cervical Spinal O.S O.3 1.1 O.9 O.3 O6 O.2 Cord Lower Cervical Spinal O.2 O.2 O.1 0.4 O.6 O.3 O.1 Cord Thoracic Spinal Cord O.1 O.1 O.1 O.3 O.3 O.2 O.OS Lumbar Spinal Cord X O.1 O.1 O.2 O.2 O.2 O.O2 Circle of Willis & Basilar 8.9 S.O 3.4 X 5.9 5.8 +1.1 Artery Carotid Artery 5.3 7.1 5.9 7.5 5.9 6.3 +0.4 Renal artery (L) 1.8 3.3 1.9 9.6 1.8 3.7 1.5 Superficial Nodes (2) 2.3 2.1 2.7 2.6 2.3 2.4 O.1 Cervical Nodes (2) 2.5 2.6 2.7 2.5 2.7 2.6 O.O Axillary Nodes (2) 2.2 1.4 1.9 4.6 3.1 2.6 O.6 Blood Sample 249.6 388.4 53.0 6.6 417.6 223.0 84.2 Muscle (R, deltoid) O.O O.9 1.2 X 1.5 O.9 O.3 Liver (R, Superficial lobe) 1.1 O.S O.6 O.S 1.5 O.8 O.2 Kidney (L, tip) 1.6 1.9 1.3 3.7 4.3 2.6 O.6 Orine 4.7 4.6 6.7 2.7 12.8 6.3 1.7 Spleen (tip) 1.4 1.5 O.8 2.9 3.4 2.0 O.S Heart O.S 0.7 O.2 O.S O.9 O6 O.1 Lung (R, top lobe) O.9 2.3 O.8 O.9 X 1.2 +0.4 Thyroid 1813 153.4 X 314.3 X 216.4 49.6 Esophagus 2.4 S.6 1.2 5.3 8.8 4.7 1.3 Trachea 1.7 1.6 O.9 3.7 X 2.0 O.6 Drug Standard CPM 6,696,942 6,103,589 7,168,742 7,168,742 7,168,742 6,861,351 +210,321 Drug Standard CPM 6,548.447 6,157,644 7,028,950 7,028,950 7,028,950 6,758,588 +176,717 Drug Standard CPM 6,631,733 5,962,084 7,513,889 7,513,889 7,513,889 7,027,097 +316,344

04.05 X-outlier removed from analysis at the site of IN IgG in the extra brain material sampled was 0.4 nM, similar drug administration, the average IgG concentrations in the to the average cortex concentration, and a rostral to caudal respiratory and olfactory epithelia were 124.509 nM and concentration gradient was observed. Similarly, a rostral to 3,191 nM respectively. A rostral to caudal gradient of 8.0 nM caudal concentration gradient (0.6 nM to 0.2 nM) was to 3.1 nM. IgG was observed in the trigeminal nerve. A observed in the spinal cord. The average concentration of similar gradient from the olfactory bulb to the anterior IgG in the dura of the brain was 8.6 nM compared to a spinal olfactory nucleus of 1.5 nM to 0.6 nM IgG was observed. The average cortex concentration of IgG after IN adminis cord dura concentration of 0.7 nM. Other tissues sampled tration was 0.5 nM. Concentrations of IgG in other brain from the Ventral skull, the pituitary and optic chiasm, regions ranged from a low of 0.3 nM in the thalamus to a contained 2.8 nM and 1.9 nM IgG respectively. high of 0.65 nM in the septal nucleus. The hippocampus was 0406. The blood concentration of IgG at the 30 min end found to contain 0.5 nM IgG. The average concentration of point was 223.0 nM. Concentrations of IgG in peripheral US 2017/0044244 A1 Feb. 16, 2017 47 organs ranged from a low of 0.6 nM in the heart to a high 0410. Results of IN and IV Delivery of the Liquid Protein of 2.6 nM in the kidney, with urine containing 6.3 nM. Preparation after 30 Min. Concentrations of IgG in the basilar and carotid arteries were similar to the concentration in the renal artery (5.8 and 0411. On average, IN administration of the IgG liquid 6.3 nM versus 3.7nM). Average concentration of IgG in the preparation resulted in lower brain concentrations than an sampled lymph nodes was 2.5 nM. Levels of IgG in tissues equivalent IV dose administered at the 30 min end point (for measured to assess variability of IN administration and example the average cortex concentration of 1.3 nM vs. 2.6 breathing difficulty (lung, esophagus, and trachea) were nM). However, to achieve these brain concentrations of IgG, fairly consistent across animals. IgG levels in the thyroid IV administration resulted in blood concentrations that were varied greatly prior to the removal of outliers. a hundred times higher than IN administration (1,361 nM. vs. 0407 Overall, IN administration of the IgG liquid prepa 13.9 nM). Higher IgG concentrations in peripheral organs ration resulted in higher brain concentrations than the micro and systems were also observed with IV vs. IN administra sphere preparation when normalizing to a 6.0 mg dose with tion. For example, IgG concentrations in the lymphatic brain concentrations ranging from 0.4 to 1.7 nM. A Summary system were ten times greater with IV vs. IN administration of the IN. IV and Fab data is presented in Table 8. This could (43.6 nM vs. 4.7 nM). be explained by lower concentrations of the microsphere IgG reaching the olfactory and respiratory epithelium. Intra 0412. When normalizing tissue concentrations to blood, nasal microsphere preparation also resulted in about ten liver, or lymphatic concentrations, it was apparent that IN times higher concentrations of IgG in the blood than the administration targets the central nervous system. The ratio liquid preparation. of tissue concentrations to blood concentrations of intranasal 0408 Normalized to a 6 mg IN dose, Fab tissue concen and intravenous IgG is presented in Table 12. For example trations were on average 19-fold higher in the brain than the for frontal cortex, IN administration results in a 48 fold liquid IgG preparations. A Summary of the tissue concen higher concentration than IV when normalizing for blood trations of IgG normalized to a 6 mg dose is presented in concentration, 40 fold higher when normalizing to liver Table 9. The three times smaller molecular weight of Fab concentration, and 5 fold higher when normalizing to aver versus intact IgG is likely responsible for the increased age lymph concentration. Intranasal administration efficiency of direct delivery from the nasal cavity to the increased IgG targeting about 50-fold more than IV admin CNS. If the Fab has similar biological effects as IgG for the istration (relative to the blood) to areas of the brain known treatment of Alzheimer's disease, it would be a promising to accumulate f-amyloid and heme (both known to bind candidate for IN delivery. IgG) including the frontal cortex, hippocampus, and the 04.09 Comparisons of brain tissue concentrations (nM) blood vessel walls of the cerebrovasculature. Importantly, after intranasal IgG liquid and microsphere preparations at B-amyloid tightly binds heme and heme is both a strong 30 and 90 min end points are depicted in FIG. 2A and FIG. pro-oxidant and pro-inflammatory agent known to inactivate 2B. brain receptors involved in memory. TABLE 12 Comparison of intranasal and intravenous targeting of IgG. Tissue to Avg. Lymph Tissue to Blood Ratios Tissue to Liver Ratios Ratios

IV IN INIV IV IN INIV IV IN INIV

Olfactory O.O32 31.649 10O2.1 O.319 266.6SO 836.2 0.986 94.481 95.8 Epithelium Respiratory O.O3O 9764.S11 323269.6 0.3OS 82267.96S 269754.1 O.943 29149.726 3O898.8 Epithelium Anterior O.OO8 0.937 122.O O.O78 7.896 1018 0.240 2.798 11.7 Trigeminal Nerve Posterior O.OOS O427 92.8 O.O46 3.600 77.5 0.144 1.276 8.9 Trigeminal Nerve Olfactory Bulbs O.OO2 O.294 119.4 O.O25 2.478 99.6 0.077 O.878 11.4 Anterior O.OO1 O.1OS 73.3 0.014 O.883 61.2 O.045 O.313 7.0 Olfactory Nucleus Frontal Cortex O.OO2 O.102 48.1 O.O22 O.863 40.1 OO67 O.306 4.6 Parietal Cortex O.OO2 O.066 26.9 O.O2S 0.556 22.5 0.077 O.197 2.6 Temporal Cortex 0.002 O.O81 38.2 0.022 O.686 31.9 OO67 O.243 3.7 Occipital Cortex 0.002 O.130 78.1 O.O17 1.093 65.2 0.052 O.387 7.5 Extra Cortex O.OO1 O.O73 S4.9 O.O13 O.611 45.8 O.041 O.217 5.3 Amygdala O.OO1 O.103 73.5 O.O14 O.867 613 O.044 O.307 7.0 Striatum O.OO1 O.OS2 39.2, O.O14 O.442 32.7 O.042 O.156 3.7 Septal Nucleus O.OO1 O.06S 48.9 O.O13 O.S49 40.8 O.042 O.194 4.7 Hypothalamus O.OO1 O.120 80.8 O.O15 1.008 67.4 O.046 0.357 7.7 Thalamus O.OO1 O.O3O 24.7 O.O12 O.254 20.6 0.038 O.O90 2.4 Midbrain O.OO1 O.049 37.7 O.O13 O411 314 O.O40 O.146 3.6 Hippocampus O.OO1 O.041 51.8 O.OO8 O.346 43.2 O.O2S O.123 S.O Pons O.OO1 O.062 48.3 0.013 O.S22 40.3 O.040 O.185 4.6 Medulla O.OO1 O.062 47.4 0.013 O.S26 39.5 O.O41 O.186 4.5 Cerebellum O.OO1 O.OS6 40.O 0.014 O.470 334 0.044 O.166 3.8 Extra Slice #1 O.OO1 O-116 77.8 0.015 O.978 64.9 O.047 O346 7.4 US 2017/0044244 A1 Feb. 16, 2017 48

TABLE 12-continued Comparison of intranasal and intravenous targeting of IgG. Tissue to Avg. Lymph Tissue to Blood Ratios Tissue to Liver Ratios Ratios

IV IN INIV IV IN INIV IV IN INIV

Extra Slice #2 O.OO2 O.O71 47.2 O.O15 O. 602 39.4 O.O47 O.213 4.5 Extra Slice #3 O.OO2 O.O59 36.5 O.O16 0.497 3O.S O.OSO O.176 3.5 Extra Slice #4 O.OO2 O.OSO 28.2 O.018 O422 23.5 O.OS6 O.149 2.7 Extra Slice #5 O.OO2 O.OS3 28.1 O.019 O451 23.5 O.O59 O-160 2.7 Extra Slice #6 O.OO2 O.O66 33.9 O.O2O 0.553 28.3 O.O60 O.196 3.2 Pituitary O.007 0.585 79.0 0.075 4.928 65.9 O.231 1746 7.5 Optic Chiasm O.004 O.S28 1418 O.O38 4.450 118.3 O-116 1577 13.6 Dorsal Dura O.O2O 1.100 54.2 O.2OS 9.268 45.2 O.634 3.284 5.2 Ventral Dura O.O17 1075 62.2 O.174 9. OS3 S1.9 O.S39 3.208 5.9 Spinal Dura O.O3S O.200 5.8 O.351 1688 4.8 1.084 O.S98 O.6 Upper Cervical O.OO1 O.O89 60.3 O.O15 O.749 S.O.3 O.046 O.26S 5.8 Spinal Cord Lower Cervical O.OO2 O.O44 23.2 O.019 0.372 19.4 O.O59 O.132 2.2 Spinal Cord Thoracic Spinal O.OO1 O.O32 27.7 O.O12 O-269 23.1 O.036 O.09S 2.6 Cord Lumbar Spinal O.OO2 O.O22 14.4 O.O16 O.188 12.0 O.049 OO67 1.4 Cord Circle of Willis & 0.013 O.837 62.8 O.13S 7.048 52.4 O416 2497 6.O Basilar Artery Carotid Artery O.O24 1.013 41.6 O.246 8.537 34.7 O.761 3.025 4.0 Renal artery (L) O.O82 O.315 3.9 O.82S 2.651 3.2 2.552 O.939 0.4 Superficial O.019 O.341 18.4 O.187 2.876 15.3 O.S8O 1.019 1.8 Nodes (2) Cervical Nodes O.046 O.398 8.7 O465 3.356 7.2 1438 1.1.89 O.8 (2) Axillary Nodes O.O31 O.26S 8.4 O.318 2.235 7.0 O.983 O.792 O.8 (2) Blood Sample 1.OOO 1.OOO 1.O 10.097 8.425 O.8 31.233 2.98S O.1 Muscle (R, O.O14 O.190 13.6 O.142 1603 11.3 O438 O.S68 1.3 deltoid) Liver (R, O.O99 O. 119 1.2 1.OOO 1.OOO 1.O 3.093 O3S4 O.1 Superficial lobe) Kidney (L, tip) O.261 O440 1.7 2.63S 3.711 1.4 8.150 1.315 O.2 Urine O.O68 O.S84 8.6 O.687 4.917 7.2 2124 1.742 O.8 Spleen (tip) O.168 O434 2.6 1.693 3.658 2.2 S.236 1296 O.2 Heart O.046 O.09S 2.0 O469 O.803 1.7 1452 O.284 O.2 Lung (R, top O.192 O.210 1.1 1936 1.769 O.9 5.990 O627 O.1 lobe) Thyroid O.393 10.607 27.0 3.963 89.366 22.5 12.259 31.66S 2.6 Esophagus O.O21 O.312 15.1 O.209 2.627 12.6 O645 O.931 1.4 Trachea O.O21 O.281 13.5 O.209 2.365 11.3 O.647 O.838 1.3

0413 Eight rats received IV IgG liquid preparation at an mean, standard error, and outliers because the blood con average dose of 6.0 mg in 47.4 uL containing 69.5 uCi centration was less than 20% of the value observed in all (diluted in saline to a total volume of 500 uL for injection) other animals, suggesting the IV infusion was not success with a 30 min end point. Animals tolerated the IV admin ful. Nanomolar concentrations of intravenously adminis istration well and all survived until the 30 min desired end tered IgG liquid preparation were measured in seven rats at point. One animal (BAX-3) was removed from analysis of the 30 min end point and presented in Table 13. TABLE 13 Tissue concentrations of intravenously administered IgG liquid preparation was measured in rats at the 30 min end point and outliers were removed.

BAX-5 BAX-7 BAX-9 BAX-10 BAX-11 BAX-13 BAX-15 Avg SE

Volume Delivered 47.0 47.0 48.0 48.0 48.0 48.0 48.0 47.7 O2 (IL) uCi Delivered 69.7 69.5 70.5 70.3 70.1 68.3 68.3 69.5 O3 mg Delivered 6.O 6.O 6.O 6.0 6.O 5.9 5.9 6.O O.O3 Olfactory 33.0 4.O.S 40.4 43.O 56.5 32.O 55.5 43.O 3.7 Epithelium Respiratory 30.4 33.5 46.7 39.1 59.5 29.0 49.4 41.1 4.3 Epithelium US 2017/0044244 A1 Feb. 16, 2017 49

TABLE 13-continued Tissue concentrations of intravenously administered IgG liquid preparation was neasured in rats at the 30 min end point and Outliers were removed. BAX-5 BAX-7 BAX-9 BAX-10 BAX-11 BAX-13 BAX-15 Avg SE

Anterior 7.4 14.8 13.5 10.3 10.1 7.9 9.2 1O.S 1.O Trigeminal Nerve Posterior 4.2 11.0 8.3 6.7 5.4 3.5 4.8 6.3 1.O Trigeminal Nerve Olfactory Bulbs 2.2 2.8 5.5 3.7 3.2 9 4.2 3.4 OS Anterior Olfactory .1 2.1 3.3 8 1.9 .2 2.2 1.9 O3 Nucleus Frontal Cortex 2.5 4.0 3.2 8 2.7 .2 4.9 2.9 OS Parietal Cortex 3.3 5.2 3.0 .6 2.6 3 6.4 3.3 O.7 Temporal Cortex 7 3.7 2.5 2.2 5.8 .5 X 2.9 O.7 Occipital Cortex 9 2.8 2.5 2.3 X 8 X 2.3 O2 Extra Cortex 4 2.1 2.6 8 X .1 X 1.8 O3 Amygdala 5 1.9 X 2.1 2.O 8 2.2 1.9 O1 Striatum 2.4 1.6 1.6 3 1.5 8 2.6 1.8 O2 Septal Nucleus 6 1.4 2.0 .6 X 2.0 2.2 1.8 O1 Hypothalamus 2 2.4 2.7 7 1.9 .5 2.7 2.0 O2 Thalamus .2 1.3 1.8 3 2.1 O.9 3.1 1.7 O3 Midbrain .1 1.4 2.3 3 2.2 .1 2.9 1.8 O3 Hippocampus 1 1.3 O6 3 X .1 X 1.1 O1 Pons 1 1.6 2.4 .4 1.6 3 2.8 1.7 O2 Medulla 2 1.4 2.7 .5 X .2 2.7 1.8 O3 Cerebellum 3 1.7 2.5 8 2.9 .2 X 1.9 O3 Extra Slice #1 5 2.8 2.7 7 2.2 3 2.1 2.0 O2 Extra Slice #2 6 3.6 2.2 .4 1.8 .2 2.7 2.1 O3 Extra Slice #3 9 3.3 2.1 .4 2.O .1 3.6 2.2 O3 Extra Slice #4 9 3.1 2.5 .4 2.6 .1 4.3 2.4 +0.4 Extra Slice #5 7 3.0 2.1 .5 3.4 .1 5.3 2.6 O6 Extra Slice #6 .9 2.4 2.2 .6 3.9 3 5.3 2.6 OS Pituitary 10.9 X 12.7 9.4 8.7 7.1 11.8 10.1 O.8 Optic Chiasm 5.9 5.2 8.4 3.9 4.7 2.9 4.6 S.1 O.7 Dorsal Dura 14.8 31.7 30.7 31.0 29.5 18.2 37.4 27.6 3.O Ventral Dura 16.4 31.0 34.4 19.6 18.9 13.8 30.4 23.5 3.1 Spinal Dura 52.3 45.9 54.8 37.6 X 53.4 39.5 47.2 3.O Upper Cervical 1.4 2.3 2.7 8 2.1 1.7 2.0 2.0 O2 Spinal Cord Lower Cervical 2.5 2.5 3.7 3.4 1.4 2.4 2.2 2.6 O3 Spinal Cord Thoracic Spinal 1.6 1.9 2.8 .1 1.O 1.2 1.3 1.6 O2 Cord Lumbar Spinal 2.8 1.8 2.5 2.0 1.3 1.3 3.1 2.1 O3 Cord Circle of Willis & 16.8 23.8 X 14.2 15.7 9.9 28.4 18.1 2.8 Basilar Artery Carotid Artery 21.7 33.8 37.2 37.5 37.4 43.9 20.6 33.2 3.3 Renal artery (L) 98.8 1292 76.5 94.4 129.3 139.0 X 111.2 10.1 Superficial Nodes 20.3 29.6 22.9 31.9 35.3 12.6 24.1 25.3 2.9 (2) Cervical Nodes (2) 32.5 39.6 78.7 43.4 83.9 65.5 94.9 62.6 9.2 Axillary Nodes (2) 103.2 31.9 75.4 18.6 37.9 14.1 18.6 42.8 12.8 Blood Sample 1,224.9 1,234.2 1,543.3 1,322.6 1,364.7 1413.4 1422.9 1,360.9 42.5 Muscle (R, deltoid) 39.06 19.5 24.6 13.3 13.0 10.7 13.4 19.1 3.8 Liver (R, 742 72.2 11S.O 126.1 122.2 1863 247.5 1348 23.7 Superficial lobe) Kidney (L, tip) 347.7 313.9 287.3 459.1 397.7 441.0 238.9 355.1 3O8 Orine 32.9 1745 1873 41.1 68.3 122.8 21.1 92.6 26.0 Spleen (tip) 234.3 241.9 196.8 317.5 232.6 175.8 198.1 2281 17.5 Heart 57.7 42.1 87.5 53.5 44.2 35.6 122.1 63.2 11.7 Lung (R, top lobe) 392.8 289.6 219.1 104.5 482.5 177.0 1614 261.0 51.3 Thyroid 317.8 651.8 832.2 S22.9 545.5 372.O 496.9 534.2 65.O Esophagus 24.8 41.3 28.0 42.8 24.4 2O.S 15.1 28.1 3.9 Trachea 14.6 29.2 17.9 39.2 13.4 59.0 24.2 28.2 6.2 Drug Standard 7,378,277 7,493,218 7,635,815 7,367,611 7,809,027 6,770,035 7,683,717 7,448,243 +128,561.8 CPM Drug Standard 7,962,330 7,709,707 6,369,627 6,846,596 6,401,005 7,249,509 7,089,796 +272,233.7 CPM Drug Standard 7,947,735 8,077,594 6,447,049 6,626.261 7,855.283 7,390,784 +351,624.3 CPM X = outlier removed from analysis

0414. The blood concentration of IgG at the 30 min end tory and olfactory epithelia were low as expected (43 nM point was 1,361 nM. Concentrations of IgG in the respira- and 41 nM respectively). A rostral to caudal gradient of 10.5 US 2017/0044244 A1 Feb. 16, 2017 50 nM to 6.3 nM IgG was observed in the trigeminal nerve. A 0421 2 mL of the pooled blood supernatant was ultra similar gradient from the olfactory bulb to the anterior centrifuged at 5,000xg (7,071 rpm) for 90 minutes at 4°C. olfactory nucleus of 3.4 nM to 1.9 nM IgG was observed. to in a 100 kDa filter tube. After the first two rats, it was The average cortex concentration of IgG after IV adminis found that 2 mL took a lot of time to filter so for and animals tration was 2.6 nM. Concentrations of IgG in other brain that followed, we centrifuged only 1 mL of the pooled blood regions ranged from a low of 1.1 nM in the hippocampus to Supernatant. At the same time, 2 mL of the pooled blood a high of 2.0 nM in the hypothalamus. The average con supernatant in the ultracentrifuged at 5,000xg (7,071 rpm) centration of IgG in the extra brain material sampled was 2.3 for 90 minutes at 4° C. to in a 30 kDa filter tube. After the nM. similar to the average cortex concentration, and a first two rats, it was found that 2 mL took a lot of time to concentration gradient was not observed. Similarly, a con filter so for and animals that followed, only 1 mL of the centration gradient was not observed in the spinal cord and pooled blood Supernatant was centrifuged. the average IgG concentration was 2.1 nM. The average concentration of IgG in the dura of the brain was 25.6 nM 0422 And 2 mL of the pooled blood supernatant was compared to a spinal cord dura concentration of 47.2 nM. ultracentrifuged at 5,000xg (7,071 rpm) for 90 minutes at 4 Other tissues sampled from the ventral skull, the pituitary C. to in a 10 kDa filter tube. After the first two rats, it was and optic chiasm, contained 10.1 nM and 5.1 nM. IgG found that 2 mL took a lot of time to filter, for subsequent respectively. animals only 1 mL of the pooled blood Supernatant was 0415 Concentrations of IgG in peripheral organs ranged centrifuged. Triplicate 25 uL samples were removed for from a low of 19.1 nM in the muscle to a high of 355.1 in gamma counting from the filtrate (100 kDa filter tube), the the kidney, with urine containing 92.6 nM. IgG concentra retentate (100 kDa filter tube), the filtrate (30 kDa filter tions in basilar and carotid arteries were considerably lower tube), the retentate (30 kDa filter tube), the filtrate (10 kDa than the renal artery (18.1 and 33.2 nM versus 111.2 nM). filter tube) for gamma counting, the retentate (10 kDa filter Average concentration of IgG in the sampled lymph nodes tube) for gamma counting. was 43.6 nM. 0423. Each brain was removed (on ice), weighed, and placed into a glass tissue homogenizer, the brain was manu Example 3 ally homogenized (40-50 passes) with homogenization buf fer at a 1:3 dilution (i.e., 2 mL buffer per g wet brain) and The Effect of IN and IV Delivery on the Intactness the homogenate was transferred into a pre-weighed conical of IgG tube (15 mL) and stored on ice. Triplicate 25 uIl samples from brain homogenate were removed for gamma counting. 0416 A study was conducted to examine whether IgG The sample was centrifuged at 1,000xg (3,160 rpm) for 10 remains intact after IN and IV administration. Specifically, minutes at 4° C. Brain Supernatant was removed into pre rats were administered 'I radiolabeled IgG either intrana weighed ultracentrifuge tube and stored on ice. Sally or intravenously and the total intact and degraded IgG was determined 30 min after administration. 0424 The extraction procedure was repeated a second time on the pellet (i.e., added same Volume of homogeni 0417 Experimental Design: The rats were anesthetized zation buffer to conical test tube containing pellet, inverted and IgG was administered as described above in Example 2. several times to dislodge the pellet, transferred into glass Blood and brain was sampled and intact IgG was detected. homogenizer, homogenized with 20-30 passes, transferred 0418 Blood was sampled approximately 30 minutes after to same pre-weighed conical test tube, centrifuged, and intranasal administration prior to perfusing with at least 100 removed Supernatant). All brain Supernatant was pooled and mL of Saline containing protease inhibitors and serum was stored in the same pre-weighed conical tube. The extraction processed. procedure was repeated a third time on the pellets. Triplicate 0419 Each blood sample (1.0 mL) was added to glass/ 25 uL samples from pooled brain Supernatant were removed tissue homogenizer containing 2.0 mL of homogenization for gamma counting. buffer (H.B., 10 mM tris buffer, pH 8.0 containing protease inhibitors) and aprotinin (100 uL per mL blood). The sample 0425 2 mL of the pooled brain supernatant was ultra was manually homogenized (30 passes) and then transferred centrifuged at 5,000xg (7,071 rpm) for 90 minutes at 4°C. into a pre-weighed conical tube (15 mL) and stored on ice. to in a 100 kDa filter tube. After the first two rats, it was Triplicate 25 uL samples were removed for gamma count found that 2 mL took a lot of time to filter, for subsequent 1ng animals only 1 mL of the pooled blood Supernatant was centrifuged. At the same time, 2 mL of the pooled brain 0420. The sample was centrifuged at 1,000xg (3,160 supernatant in the ultracentrifuged at 5,000xg (7,071 rpm) rpm) for 10 minutes at 4°C. Blood supernatant was removed for 90 minutes at 4° C. to in a 30 kDa filter tube. After the into a pre-weighed ultracentrifuge tube and stored on ice. first two rats, it was found that 2 mL took a lot of time to The extraction procedure was repeated on the blood pellet a filter, for subsequent animals only 1 mL of the pooled blood second time (i.e. same Volume of homogenization buffer Supernatant was centrifuged. And 2 mL of the pooled brain added to conical test tube containing pellet, inverted several supernatant was ultracentrifuged at 5,000xg (7,071 rpm) for times to dislodge the pellet, transferred into glass homog 90 minutes at 4° C. to in a 10 kDa filter tube. After the first enizer, homogenized with 15 passes, transferred to same two rats, it was found that 2 mL took a lot of time to filter, pre-weighed conical test tube, centrifuged, and blood Super for subsequent animals only 1 mL of the pooled blood natant removed). All blood Supernatant was pooled and Supernatant was centrifuged. stored in the same pre-weighed conical tube. The extraction procedure was repeated on the blood pellet a third time. 0426 Triplicate 25uL samples were removed for gamma Triplicate 25 uL samples from pooled blood supernatant counting from the filtrate (100 kDa filter tube), the retentate were remove for gamma counting. (100 kDa filter tube), the filtrate (30 kDa filter tube), the US 2017/0044244 A1 Feb. 16, 2017

retentate (30 kDa filter tube), the filtrate (10 kDa filter tube) were made to include /3 of animals in each size group. Mice for gamma counting, the retentate (10 kDa filter tube) for were re-evaluated to make new size groups every two gamma counting. weeks. The mice were divided into five treatment groups of 0427. Results: Two rats received IV IgG liquid prepara 20 mice as described in Table 15. tion and two rats received IN IgG liquid preparation at an average dose of 52 LL containing 56 uCi (diluted in saline TABLE 1.5 to a total volume of 500 uL for IV injection) with a 30 min end point. Animals tolerated the administration well and all Treatment groups assigned for intranasal administration of IgG. survived until the 30 min desired end point. Mouse Strain Drug Administration Description 0428. In the brain, approximately 80% of gamma counts Tg2576 IN IgG 0.4 g/kg/2 wk “TG-High' from '''I-labeled IgG after both IN and IV delivery were Tg2576 IN IgG 0.8 g/kg/2 wk TG-Low' greater than 100 kD. Suggesting intact protein. In the blood, Tg2576 IN Saline (control) “TG-Saline 100% gamma counts from I-labeled IgG after IV delivery C57 IN IgG 0.8 g/kg/2 wk “WT-High' were greater than 100 kD, suggesting all was intact. With IN C57 IN Saline (control) WT-Saline delivery, only 33% of gamma counts from I-labeled IgG found in blood was greater than 100 kD. Suggesting that 0431. The mice were ordered and received in the animal 'I-labeled IgG may be broken down and enter the blood as facility at 2 months of age and were singly housed and aged part of the clearance process from the nasal cavity, the brain for 6 months. At 8 months of age, the mice were acclimated or both. This also provides additional evidence that IgG to handling for awake intranasal delivery over the course of entering the CNS after IN administration does not travel 1 month. Mice were then intranasally treated with IgG or from the nasal cavity to the blood to the brain, but rather saline three times/week for 7 months. At 16 months of age, along direct pathways involving the olfactory and trigeminal behavioral testing occurred for 5 weeks while intranasal nerves. A Summary of the intactness of IgG in the brain and treatment continued. At ~17 months of age, 12 mice/group blood after intranasal or intravenous administration is pre were euthanized and brain tissue was collected for analysis. sented in Table 14. 0432 IgG and saline for IN delivery was prepared Friday afternoons from stocks sent by Baxter, and stored at ~4° C. TABLE 1.4 for use the following week. Solutions were made to deliver Summary of Intactness of IgG in the Brain and Blood. a dose of either 0.4 mg/kg/2 wk IgG or 0.8 mg/kg/2 wk IgG, and were made to deliver a total of 24 uL. Drug was also IN IV made to cater to each of the three size classes within a treatment group. R1 R3 Avg R2 R4 Avg 0433 Mice were acclimated to handling for a period of BLOOD two-four weeks before the onset of intranasal dosing. Accli mation to handling was important, as it helped ensure a % greater than 100 kD 30 36 33 123 113 118 % greater than 30 kD 34 34 34 123 110 116 correct body position for maximum effectiveness of awake % greater than 10 kD 67 57 62 99 108 104 intranasal drug delivery. In addition, mice that have not been BRAIN properly accustomed to this process can have a severe anxiety reaction after dosing. Mice spent about 1-3 days on % greater than 100 kD 93 70 81 78 77 77 % greater than 30 kD 87 78 82 83 84 83 each of nine steps before progressing to the next step, % greater than 10 kD 88 78 83 88 93 91 depending upon the animals comfort to handling. The mouse's stress level was used as a measure of progress. This means monitoring the mouse's movements, the amount/ frequency of urination, defecation, trembling, and biting. If Example 4 a mouse had a high stress response, it remained on that step before progressing to the next until the response is reduced. Effect of Intranasal Administration of IgG on A sample acclimation schedule can be seen in Table 16. Amyloid Plaque Loads Acclimation of the mice progressed through the following 0429. A study was conducted to examine whether intra once-a-day steps. The steps were not performed more than nasal administration of IgG decreases amyloid plaque loads once per day in order to minimize the anxiety in the mice. in the brain in vivo. The purpose of the study was to 0434 First, the mouse was placed in the palm of the hand determine whether chronic treatment with intranasally deliv for a period of two to three minutes, no more than one foot ered IgG at two doses (0.4 g/kg/2 wk and 0.8 g/kg/2 wk) above the cage top, as animals frequently jumped during this would have any effect on the amyloid plaque load in a introductory step. If the mouse attempted to crawl out of the transgenic amyloid mouse model of Alzheimer's disease. hand and up one's arm, the mouse was lifted by the base of 0430. Experimental Design: The TG2576 (“TG”) amy the tail and placed back in one's hand. Second, the mouse loid mouse model was used in this study as a mouse model was placed in the palm of the hand for three minutes and for Alzheimer's disease and C57 mice were used as controls. petted gently. The mouse was petted directionally from head TG2576 mice (cat. #1349-RD1-M) were acquired from to tail, while allowing the animal to move about freely. Taconic, Inc. in two batches of 50 spaced one month apart Third, the mouse was placed in the palm of the hand for (Batch 1 and Batch 2). Animals were individually housed three minutes while massaging behind the ears (lightly with free access to food and water, and were kept on a 12 pinching together the skin on the back of the neck using the hour light cycle. For dosing with drug in a mg/kg dosing thumb and pointer finger). Fourth, the mouse was held/lifted scheme, mice were divided into three size classes within by the scruff of its neck for 30 seconds, letting the mouse rest each treatment group, Small, medium, and large. Size groups on the cage top for 30 seconds before repeating the hold US 2017/0044244 A1 Feb. 16, 2017 52 again. Fifth, the mouse was held using the intranasal grip, (IHC) for amyloid plaques, inflammatory markers, and without inverting the animal, for a period of 30 seconds and soluble and insoluble amyloid. then released back to the cage top. This was repeated a 0438 Prior to euthanasia via transcardial perfusion, mice second time after a one-minute rest period. Sixth, the mouse were anesthetized with sodium pentobarbital (60 mg/kg was held with the intranasal grip while inverting the animal diluted 1:4 with sterile saline). A first booster of half the full So its ventral side was facing up towards the ceiling with the dose was given followed by additional quarter-dose boost animal's neck is parallel to the floor. This position was held ers, if necessary. The level of anesthesia and sensitivity to for 30 seconds and was then repeated a second time after a pain was monitored every five minutes throughout the one-minute rest period. If the mouse freed itself from the procedure by testing reflexes including pinching the hind grip, the mouse was put back on the cage top and re-gripped. paw and tail. Mice were then euthanized with transcardial If the mouse's stress level increased too much, the mouse perfusion with 15 ml ice cold saline (no protease inhibitor was returned it to the cage. Seventh, the mouse was held needed) and blood was collected from the heart. Briefly, the with the intranasal grip, inverted and a pipettor with an arms of the mouse were taped down. The skin was cut to empty tip was briefly placed over each nostril for 30 expose the sternum. A hemostat was used to hold the seconds. This step was repeated after a one-minute rest Sternum while blunt dissection Scissors were used to cut period. Eighth, the mouse was held with the intranasal grip, vertically on both sides of the sternum making an incision inverted, and intranasally administered 6 Jul of saline into the with a V-shape to expose the heart. Blood was collected left and right nare. Ninth, the mouse was held with the from the heart prior to perfusion and processed into serum. intranasal grip, inverted, and intranasally administered 6 ul A Small hole in the left ventricle was made using a 24-gauge of saline into the left and right nare twice placing the animal cannula. The cannula was inserted into the aorta and held in back on the cage top in between. place. Extension tubing (filled with 5 mL of 0.9% NaCl) was attached to the cannula and the animal was manually per TABLE 16 fused with 15 ml saline. 0439 Blood was spun down and serum divided into two Sample schedule for acclimation to awake IN drug delivery. aliquots. One aliquot was 50 uL and will be eventually pooled and sent for analyses of overall health of the treat Day # Day Action ment group. The remaining serum was placed into its own 1 M Hold for -2-3 min tube and Snap frozen for other analyses. 2 Tu Hold for -2-3 min 3 W Hold and pet -2-3 min 0440 The brain was collected and hemisected Sagitally in 4 T Hold and pet -2-3 min amouse brain matrix. The left half of the brain was dissected 5 F Lightly pinch scruff into olfactory bulbs, cortex/hippocampus mix, septum, mid 6 M Lightly pinch scruff brain/diencephalon, brainstem (down to the V of the upper 7 Tu Scruff and lift 8 W Scruff and lift cervical spinal cord), and cerebellum. These tissues were 9 T intranasal Grip placed into microcentrifuge tube and Snap frozen in liquid 10 F intranasal Grip nitrogen. The right half was left in the matrix and sliced 3 11 M intranasal (IN) Grip and Invert mm from the centerline. The inner portion towards the 12 Tu intranasal (IN) Grip and Invert 13 W N Grip, Invert, empty pipette tip center of the brain was post-fixed in formalin (in a 15 ml 14 T N Grip, Invert, empty pipette tip conical tube filled with 10 ml formalin) and sliced for IHC 15 F N Grip, Invert, deliver 1 round saline to each nare analyses. The outer portion was Snap frozen in liquid nitro 16 M N Grip, Invert, deliver 1 round saline to each nare gen for eventual analysis for inflammation. 17 Tu N Grip, Invert, deliver 2 rounds saline to each nare 18 W N Grip, Invert, deliver 2 rounds saline to each nare 0441 The medial 3 mm sagittal section of the right half of the mouse brain was fixed by placing them each into 20 mL of 10% formalin. These samples were fixed for several 0435 For awake intranasal delivery of drug, the intrana hours at room temperature and then overnight at 4° C. on sal grip, each mouse was restrained twice and held with their slow moving rocker. The fixed Sagittal brain sections were necks parallel to the floor while a volume of 24 ul of liquid placed medial side down into labeled pathology cassettes. was administered. Specifically, un-anesthetized mice were The pathology department at Region’s Hospital conducted grabbed by the scruff of their necks and held gently, but the paraffin processing and embedding (dehydrate, infiltrate firmly, in an inverted position so that the mouse cannot move with paraffin, mount into paraffin blocks). The paraffin around. Each mouse was given four 6 ul nose drops (alter blocks were blinded by coding/relabeling. nating nares) using a 20-ul pipettor. Intranasal drug delivery 0442. The paraffin blocks were sectioned at a thickness of began when mice were 9 months of age. 5um using the Leica RM2235 microtome and collected on Superfrost Plus microscope slides (Cardinal Health, cathi 0436. At 16 months of age, mice were subjected to a five M6146-PLUS). Seven sections were collected per mouse, week battery of behavioral tests to assess for memory, with at least/approximately 100 um of tissue removed sensorimotor, and anxiolytic changes. These included Mor between tissue section collections (labeled 1-6 from, medial ris water maze hidden and visual platform tests (reference to lateral). To increase the quality of the sections to be memory, visual ability), radial arm water maze (working stained, a dissection microscope was used to identify and memory), passive avoidance task (memory), Barnes maZe remove one of the seven sections. (memory), open field test (exploratory behavior), elevated 0443 Slides were deparaffinized and hydrated. Specifi plus maze (anxiety), and rotarod (motor skills). cally, the slides were placed in a glass staining jar rack for 0437. After behavior, 12 mice from each treatment group easy transfer between staining dishes. The paraffin wax was were euthanized and their brains collected for biochemical removed with Xylene washes (clearing) and then hydrated analyses. These analyses include immunohistochemistry with ethanol/water. Specifically, the slides were washed in US 2017/0044244 A1 Feb. 16, 2017

xylene three times for five minute intervals, washed in 100% analysis and the threshold function to select a minimum ethanol two times for five minute intervals, washed in 95% pixel value that defined each identified particle as qualifying ethanol one time for five minutes, rinsed in running water for as a plaque. These values were determined by analyzing five minutes, and rinsed in PBS for five minutes. multiple positive and negative controls and verifying which 0444 Heat induced epitope retrieval (HIER) was used to values correctly identified the plaques in these control slides. pretreat the slides prior to antibody staining. A Tris/EDTA The region of interest within each image was chosen by a Buffer (pH 9) was used. The slides were immersed in a blinded researcher who was instructed to place the region of steamer proof dish containing the Target Retrieval Solution interest in the position that would maximize the inclusion of (Tris/EDTA pH 9) pre-warmed to 70° C. The dish with slides plaques. The size (pixels) and number of plaques identified was then placed in the steamer and incubate for 30 minutes were copied into excel for data analysis. The plaques were at 97° C. The steamer was turned off and allowed to cool to then characterized by their relative size. The plaque sizes at least 65° C. The container of slides was removed from the reported in this study refer to the calculated radius of a steamer and allowed to cool for another 10-15 minutes. The plaque assuming the particle conformed to the shape of a slides were then removed from the container and rinsed in perfect circle. The number of plaques and percent area PBS for 10 min in a coplan jar. covered by plaques calculated from each region of interest 0445. Non-specific binding sites were then blocked with was used as a single data point in comparing the treatment normal serum blocking solution (300 uL/slide) for 1 hour in groups. Two tailed t-tests were used to assess the signifi a humidity chamber. Sections were incubated in a humidbox cance between groups. with primary antibody against amyloid (purified Anti-Beta 0450 Prior to staining the complete set of collected tissue Amyloid, 17-24 (4G8) Monoclonal Antibody, from Covance sections, an initial verification of the staining and micros (SIG-39220)) at a 1:200 dilution in primary antibody dilu copy analysis was conducted with relevant staining controls. tion buffer (0.01 M PBS pH 7.2) for 1 hour at room These controls included, a positive control using sections temperature. Sections were incubated in secondary antibody from one of the transgenic mice receiving saline, negative (Goat anti-mouse IgG, Alexa Fluor 647 (2 mg/ml) from controls in which either the primary or secondary antibody Invitrogen (A21235)) dilution buffer (0.01 M PBS, pH 7.2) incubation was omitted from the staining procedure and a with a 1:200 secondary concentration for 1 hr at room negative control using sections from one of the wild-type temperature. mice receiving saline. Additional controls, including the 0446. Slides were then counterstained with DAPI. titration of primary and secondary antibodies and the com Diluted 300 nM DAPI in PBS was used. 1 ul of 14.3 mM parison of different epitope retrieval methods have been DAPI stock was diluted into 48 ml PBS, Vortexed, and mix conducted previously in our lab using these antibodies and thoroughly. The DAPI solution was poured into coplan jar the same experimental procedure. containing the slides. The slides were incubate for 20 min at 0451 Tissue supernatants were analyzed using kits from RT. The slides were rinsed quickly in PBS, then 2x10 min Life Technologies (formerly Invitrogen; Carlsbad, Calif.; in washing buffer, followed by a 10 min incubation in PBS. part is KHB3482 (AB40) and KHB3442 (AB42)). Gener 0447 Immediately after staining, the slides were then ally, the proper dilutions were first determined with three dehydrated, cleared, and mounted. Specifically, the slides samples from either TG or WT mice, and then all samples were incubated in 95% ethanol for 5 minutes, 100% ethanol were run at that dilution. Samples were quantified using a for two five minute increments, xylene for three five minute polynomial equation fit to a standard curve. Quantities of AB increments, and mounted with a coverslip in DPX without measured in the wells were then corrected for dilutions and letting the specimen dry. The mounted slides were stored at total protein (as determined by a BCA assay). room temperature. 0448. Images of the fluorescently stained plaques were 0452 Results: captured with the AZ 100 Multizoom Macroscope with the 0453 Immunohistochemical measurement of amyloid C1 si Spectral Confocal attachment and an AZ Plan Apo 4x plaques in brain tissue slices demonstrated that there was a objective. Initial localization and focusing of the hippocam significant drug effect. Both groups of TG mice administered pus and cortex was conducted through epifluorescence IgG intranasally had significantly decreased plaque loads in imaging using filters for the DAPI stain. The scope was then the cortex (FIGS. 3A, 3B, and 3E). Switched to confocal imaging using the 637 nm laser for 0454 Nasal administration of both the low dose and high acquisition of the IHF-labeled amyloid. Fine tuning within dose of IgG significantly reduced the total percent area the Z-axis for optimal signal detection was confirmed with a covered by plaques in the cortex of TG2576 mice (FIG. 3A). 512x512 pixel resolution. Images were then captured at The percent area covered by plaques decreased by 25.7% 1024x1024 with the Nikon EZ-C1 Software and the raw (low dose; p=0.014) and 24.3%, (high dose; p=0.037), image files were saved in Nikon’s “...ids' file format. Cor respectively. The change in the percent area covered by responding tiff files of the 637 nm channel were generated plaques was slightly more pronounced at 27.1% for the low using Fiji (Image.J). The tiff files were then converted to 8-bit dose and 26.0% for the high dose when the minimum images (from 16-bit) and the contrast was enhanced by 0.5% threshold for defining a plaque was increased from a radius through batch processing (Macro programming) in Fiji of 25um to 50 um (p values of 0.01 and 0.026, respectively). (Image.J). The decrease in plaque load was also found to be significant 0449 Plaques were quantitated in selected regions of when the minimum threshold was set at 100 Lum (p values of interest in the hippocampus and cortex by determining the 0.035 and 0.021, respectively). A change in the percent area average number of plaques detected in each region and by covered by plaques was not apparent when the Smaller determining the percent area covered by plaques within each plaques (less than 50 um radius) were used exclusively in the region. Image processing and analysis was conducted in Fiji. analysis. Thus, plaque reduction in the cortex appears to be Plaques were defined within Fiji by using the particle more pronounced plaques larger than 50 um. US 2017/0044244 A1 Feb. 16, 2017 54

0455 The number of plaques in the cortex of both low 0463 Standards and samples were run in duplicate. The dose and high dose IgG treatment groups showed a trend samples and standards were run in a protease inhibitor toward a decrease in the numbers of plaques detected (FIG. cocktail with 1 mM AEBSF (a serine protease inhibitor). 3B). This decrease reached significance in the low dose IgG AEBSF was important because serine proteases can rapidly treatment group when Small plaques (less than 50 Lim radius) degrade AB peptides. The samples were kept on ice until were not included in the analysis. Specifically, treatment they were ready to be applied to the ELISA Plate. with intranasally administered IgG provided a significant 0464 Sample matrix has a dramatic impact on AB recov reduction in plaque load when the data were analyzed by ery. To ensure accurate quantitation, the standard curves inclusion of plaques having a radius of from 50 um to 100 were generated in the same diluent as the samples. A um, greater than 100 um, and greater than 50 um. The standard reconstitution buffer was prepared by dissolving decrease in plaque load reached significance for the high 2.31 grams of sodium bicarbonate in 500 mL of deionized dose IgG treatment group when the radius of analyzed water and the pH was adjusted using 2 N sodium hydroxide plaques was set at greater than 100 um. until the pH was 9.0. 0456. In contrast to the results seen in the brain cortex, 0465. The standards for a quantitative standard curve IgG treatments did not result in a significant change in either were prepared. The Hu AB42 Standard was used. The Hu the percent area covered by plaques or the numbers of Af342 Standard was allowed to equilibrate to room tempera plaques detected in the hippocampus (FIGS. 3C and 3D). ture (RT) and then reconstituted to 100 ng/mL with Standard Although intranasal administration of both low and high Reconstitution Buffer (55 mM sodium bicarbonate, pH 9.0). dose IgG appeared to result in a slightly reduced plaque load The standard mixture was Swirled and mixed gently and in the hippocampus, the reduction was minimal and did not allowed to sit for 10 minutes to ensure complete reconsti reach significance in this region of the brain. tution. The standard was then briefly vortexed prior to 0457 Immunofluorescent staining of amyloid plaques in preparing standard curve. Generation of the standard curve the hippocampus and cortex of aged TG mice is depicted in using the AB peptide standard was performed using the same FIG. 3E. As show, there is a decrease staining for amyloid composition of buffers used for the diluted experimental plaques in the hippocampus and cortex in mice that were samples. 0.1 mL of the reconstituted standard was added to treated with low and high IgG doses compared to TG mice a tube containing 0.9 mL of the Standard Diluent Buffer and treated with saline. labeled as 10,000 pg/mL. Hu AB40. The standard was mixed and then 0.1 mL of the 10,000 pg/mL standard was added to Example 5 a tube containing 1.9 mL Standard Diluent Buffer and labeled as 500 pg/mL. Hu AB40. Mix. The standard was Effect of Intranasally Administered IgG on Soluble mixed and then 0.15 mL of Standard Diluent Buffer was and Insoluble AB40 and AB42 added to each of 6 tubes labeled 250, 125, 62.5, 31.25, 15.63, 7.81, and 0 pg/mL. Hu AB40 to make serial dilutions 0458. A study was conducted to assess the efficacy of of the standard. chronic intranasal (IN) administration of IgG at two doses in 0466. The samples were then prepared for the plate. a transgenic amyloid mouse model. Specifically, measure Specifically, the samples were remove from the freezer, ments of the soluble and insoluble amyloid beta peptides allowed to thaw, and diluted to the desired dilution using Af340 and AB42 were taken in wild type and Tg2576 dilution buffer provided with the kit mixed with a protease (amyloid mouse model) pre- and post-intranasal IgG admin inhibitor tablet. The samples were kept on ice until loaded istration. The purpose of the study was to determine whether into the wells on the plate. chronic treatment with intranasally delivered IgG at two 0467. The plates were labeled as being either AB40 or doses (0.4 g/kg/2 wk and 0.8 g/kg/2 wk) would have any AB42 with a sharpie. 50 ul of standards and sample were effect on the amyloid plaque load in a transgenic amyloid added to the pre-labeled wells. 50 uL of Hu AB40 or AB42 mouse model of Alzheimer's disease. Detection Antibody solution provided with the kit was added 04.59 Experimental Design: to each well. The plate was covered and incubated for 3 0460. As described in Example 4, the TG2576 (“TG') hours at room temperature with shaking. Shortly before the amyloid mouse model was used in this study as a mouse 3 hours expired, the Anti-Rabbit IgG HRP Working Solution model for Alzheimer's disease and C57 mice were used as was prepared. To make this, 10 uL of Anti-Rabbit IgG HRP controls. The handling of the mice, preparation of drug, and (100x) concentrated solution was diluted in 1 mL of HRP administration of drug was conducted as described above in Diluent for each 8-well strip used in the assay and labeled as Example 4. Anti-Rabbit IgG HRP Working Solution. 0461 The mice were divided into five treatment groups 0468. The solution was thoroughly decanted from wells of 20 mice as described in Table 15. At approximately 17 and the wells were washed 5 times with 300 uL of wash months of age and 12 months of treatment, 12 mice from solution. The plates were banged hard on lab bench to be each treatment group were euthanized and the concentration sure it was dry. 100 uL of the Anti-Rabbit IgG HRP working of the AB40 and AB42 amyloid peptides in the brains of the solution was added to each well. The plate was covered and TG and control mice were measured by ELISA to determine allowed to sit at room temp for 30 min. The solution was whether amyloid plaque concentrations changes could be thoroughly decanted from wells and the wells were washed detected. 5 times with 300 uL of wash solution. The plates were 0462 AB40 and AB42 were measured by ELISA using banged hard on lab bench to be sure it was dry. 100 uL of Invitrogen ELISA kits. The ELISA kits were stored in Stabilized Chromogen was added to each well and the plate refrigerator until they were ready to use. The kits were was immediately placed in the dark and allowed to sit for 20 removed from refrigerator and allowed to warm to room min. 100 uL of Stop Solution was added to each well and the temperature before use. sides of the plate were gently tapped to mix. US 2017/0044244 A1 Feb. 16, 2017

0469. The absorbance of each well was read at 450 nm ELISA plate based on the standard curve. These values were having blanked the plate reader within 30 minutes after then corrected for dilution of supernatant, dilution from the adding the Stop Solution. The concentrations were deter extraction process, and then given a correction factor from mined using the standard curve. a BCA analysis of total protein extracted. For each protein, 0470 Results: between 1 and 4 samples were excluded for either being 0471. The ELISA plates for both AB40 and AB42 pur statistical outliers or being too high/low to fit within the chased from Invitrogen yielded consistent standard curves. standard curve. A summary of the soluble and insoluble The best dilutions of brain supernatant for samples for Af340 concentrations are presented in Table 17 and Table 18. soluble AB40 and AB42, and insoluble AB40 and AB42 were A summary of the soluble and insoluble AB42 concentra 10x, undiluted, 10000x, and 2500x, respectively. Brain tions are presented in Table 19 and Table 20. The ratios of concentrations of each protein were analyzed by first deter soluble AB40/A342 are provided in Table 21 and the ratios mining the concentration of the sample in the well in the of insoluble AB40/A342 are provided in Table 22. TABLE 17 Soluble AB40 detected in brain. Mouse sac Date Concentration Mouse sac Date Concentration order # Group measured (pg/ml) order # Group measured (pg/ml)

1 TG-Low -Oc 9078 33 TG-Saline 1-Oc 3940 6 TG-Low -Oc 3964 38 TG-Saline 1-Oc 1328 11 TG-Low -Oc 3110 43 TG-Saline 1-Oc 1983 16 TG-Low -Oc 2788 48 TG-Saline 1-Oc 3656 21 TG-Low -Oc 3934 53 TG-Saline 1-Oc 66SO 26 TG-Low -Oc 3747 58 TG-Saline 1-Oc 61.59 31 TG-Low -Oc 3796 4 WT-High 9-Oc O 36 TG-Low -Oc 5450 9 WT-High 9-Oc O 41 TG-Low 27-Sep 5261 14 WT-High 9-Oc O 46 TG-Low -Oc 2082 19 WT-High 9-Oc O 51 TG-Low -Oc 252O 24 WT-High 9-Oc O 56 TG-Low -Oc 9448 29 WT-High 9-Oc O 2 TG-High -Oc 3061 34 WT-High 9-Oc O 7 TG-High -Oc 1814 39 WT-High 9-Oc O 12 TG-High -Oc 4681 44 WT-High 9-Oc O 17 TG-High -Oc 2509 49 WT-High 9-Oc O 22 TG-High -Oc 7869 S4 WT-High 9-Oc O 27 TG-High -Oc 6363 59 WT-High 9-Oc O 32 TG-High -Oc S541 5 WT-Saline 9-Oc O 37 TG-High 27-Sep S190 10 WT-Saline 9-Oc O 42 TG-High -Oc 3609 15 WT-Saline 9-Oc O 47 TG-High -Oc 1122 2O WT-Saline 9-Oc O 52 TG-High -Oc 12163 25 WT-Saline 9-Oc O 57 TG-High -Oc 1SO2 30 WT-Saline 9-Oc O 3 TG-Saline 27-Sep 3708 35 WT-Saline 9-Oc O 8 TG-Saline 1-Oc 4833 40 WT-Saline 9-Oc O 13 TG-Saline 1-Oc 1673 45 WT-Saline 9-Oc O 18 TG-Saline 1-Oc 4039 50 WT-Saline 9-Oc O 23 TG-Saline 1-Oc 2373 55 WT-Saline 9-Oc O 28 TG-Saline 1-Oc 41.33 60 WT-Saline 9-Oc O

Average Std deviation Std error

TG-Low 4598.418 2395.218 6914399 TG-High 3932.644 1782.644 630.2598 TG-Saline 37O6.334 1570.737 473.595 WT-High O O O WT-Saline O O O

TABLE 18 Insoluble AB40 detected in brain. Date Concentration Concentration Mouse Date Concentration Concentration Mouse sac order # Group measured (pg/ml) (ugml) sac order # Group measured (pg/ml) (ugml)

1 TG-Low 8-Oct 102571.99 10.26 33 TG- 8-Oct 25512730 25.51 Saline 6 TG-Low 8-Oct 11697779 11.70 38 TG- 8-Oct 1941498O 1941 Saline 11 TG-Low 8-Oct 7575663 7.58 43 TG- 8-Oct 26032S47 26.03 Saline 16 TG-Low 8-Oct 8322854 8.32 48 TG- 8-Oct 39277004 39.28 Saline

US 2017/0044244 A1 Feb. 16, 2017 58

TABLE 20-continued Insoluble AB42 detected in brain. WT-High O.O.35956 O.O24034 O.OO7247 WT-Saline O.OO6447 OOO4669 O.OO1477

TABLE 21 TABLE 22-continued Ratios of soluble AB40/AB42. Ratios of insoluble AB40/AB42. Mouse 2 TG-High O44986 34 WT-High 0.66754 Mouse sac Ratio of S80 Ratio of 7 TG-High O.17399 39 WT-High 0.798.17 order # Group AB42/AB40 order # Group AB42, AB40 12 TG-High 0.5557 44 WT-High 0.5905 17 TG-High O.36116 S4 WT-High 1.26484 1 TG-Low O.16O29S 33 TG-Saline O.158812S 22 TG-High O.21629 59 WT-High 1.OOS66 6 TG-Low O.13894 38 TG-Saline O.296O768 27 TG-High O.17166 5 WT-Saline 1.0335 11 TG-Low O. 164329 43 TG-Saline O.2834986 32 TG-High O.14659 10 WT-Saline 16 TG-Low O.266987 48 TG-Saline O.1180902 37 TG-High O.29242 15 WT-Saline O.88232 21 TG-Low O.1791.96 S3 TG-Saline O.19472 42 TG-High 0.22767 2O WT-Saline O.35914 26 TG-Low O.16629 S8 TG-Saline O.2209363 47 TG-High O.25291 25 WT-Saline 31 TG-Low O. 1219 4 WT-High O 52 TG-High O.32982 30 WT-Saline O.92744 36 TG-Low O. 111683 9 WT-High 57 TG-High O.15195 35 WT-Saline 41 TG-Low O.297232, 14 WT-High 3 TG-Saline O.24128 40 WT-Saline O.824O7 46 TG-Low O.290904. 19 WT-High 8 TG-Saline O.2226 45 WT-Saline 1.06434 51 TG-Low O.327174 24 WT-High 13 TG-Saline O.32O33 50 WT-Saline O.958.64 56 TG-Low O. 161546 29 WT-High 18 TG-Saline 0.3O277 55 WT-Saline 1.02571 2 TG-High O. 1893.14 34 WT-High 28 TG-Saline O.15252 60 WT-Saline 1.10371 7 TG-High O.245964 39 WT-High 12 TG-High O.188097 44 WT-High Average Stod deviation Std error 17 TG-High O.163459 49 WT-High 22 TG-High O.152272 S4 WT-High TG-Low O.3418.191 O.15905 O.04591 27 TG-High 0.13377 59 WT-High TG-High O.2727456 O.132S6 O.04.192 32 TG-High 0.1582O1 5 WT-Saline TG-Saline O.2667446 O.O6933 O.O2001 37 TG-High O.283163 10 WT-Saline WT-High 0.7461702 O.28.933 O.O8724 42 TG-High O.243714 15 WT-Saline WT-Saline O.9087633 O.22479 O.O7493 47 TG-High O.258574 20 WT-Saline 52 TG-High 0.250769 25 WT-Saline 57 TG-High O.2S651 30 WT-Saline 0472. The most obvious and expected result was that both 3 TG-Saline O.2132 35 WT-Saline 8 TG-Saline O.204815 40 WT-Saline soluble and insoluble AB40 and AB42 were drastically 13 TG-Saline O.335658 45 WT-Saline higher in all TG mice than WT mice. Soluble AB40 and 18 TG-Saline O.181432 SO WT-Saline Af342 were not detectable in WT mice, while insoluble AB40 23 TG-Saline O.219518 SS WT-Saline and AB42 were present, though at about 1000 times lower 28 TG-Saline O.1783 60 WT-Saline than in TG mice. The next most obvious result was that in all TG mice, the concentration of insoluble AB40 and AB42 Average Std deviation Std error was much higher than soluble AB40 and AB42, roughly TG-Low O.198873 O.O7SOO8 O.O217 about 5000 and 7500 times higher, respectively. TG-High O.O157 0473 Regarding group comparisons among the three TG TG-Saline O.217088 O.061325 O.O177 groups, there were no significant differences among any of WT-High O O O the groups for either soluble or insoluble AB40 or AB42 WT-Saline O O O using an ANOVA. This was somewhat surprising for insoluble amyloid as there were clear differences in plaques in the cortex between drug-treated and saline-treated TG TABLE 22 mice. The most likely explanation is that the ELISA was not as sensitive to these differences as the IHC slides of plaques. Ratios of insoluble AB40/AB42. Mouse Example 6 S80. Ratio of Mouse sac Ratio of order # Group AB42, AB40 order # Group AB42, AB40 Effect of Intranasal Administration of IgG on Weight and Survival 1 TG-Low O438O6 33 TG-Saline O.14524 6 TG-Low O.62584 38 TG-Saline O.31072 0474. A study was conducted to assess the efficacy of 11 TG-Low O.34875 43 TG-Saline O.26396 16 TG-Low O.19763 48 TG-Saline O.31298 chronic intranasal (IN) administration of IgG at two doses in 21 TG-Low 0.37995 53 TG-Saline O.3551.6 a transgenic amyloid mouse model. The purpose of the study 26 TG-Low O.13045 58 TG-Saline O.30663 was to determine whether chronic treatment with intrana 31 TG-Low O.366.82 4 WT-High O41437 sally delivered IgG at two doses (0.4 g/kg/2 wk and 0.8 36 TG-Low O.21269 9 WT-High 1.10685 g/kg/2 wk) would have any effect on the mouse weight and 41 TG-Low O.22742 14 WT-High O.80589 survival. 46 TG-Low O.2882 19 WT-High O.62142 51 TG-Low O.25915 24 WT-High 0.30555 0475 Experimental Design: 56 TG-Low O62688 29 WT-High O.62708 0476. As described in Example 4, the TG2576 (“TG') amyloid mouse model was used in this study as a mouse US 2017/0044244 A1 Feb. 16, 2017 59 model for Alzheimer's disease and C57 mice were used as TABLE 23 controls. The handling of the mice, preparation of drug, and administration of drug was conducted as described above in T-tests used to evaluate results of behavioral studies in wild type and Example 4. Alzheimer's disease mouse models administered IgG intranasally. 0477 The mice were divided into five treatment groups Comparison Reason for Comparison of 20 mice as described in Table 15. The weight and survival WT-saline vs. TG-Saline To determine whether there is a of the mice were monitored for 103 weeks. The weight of ransgenic effect of the model. each mouse was recorded weekly (data not shown). WT-all vs. TG-all (all = saline and To provide a larger scale view of IN IgG) he transgenic effect of the model. 0478 Results: These experiments showed that intranasal TG-saline vs TG-low dose IN IgG To determine whether TG mice IgG increases the lifespan of TG mice. FIG. 4A shows that reated with the low dose of IgG performed differently than TG mice TG mice have an increased lifespan when they are admin reated with saline. istered a high (0.8 g/kg/2 wk) or a low (0.4 g/kg/2 wk) dose TG-saline vs TG-high dose IN IgG To determine whether TG mice of intranasal IgG compared to TG mice administered saline reated with the high dose of IgG intranasally (control). FIG. 4B shows that TG mice admin performed differently than TG mice istered intranasal IgG had longer lifespans than WT mice. reated with saline. Although this study begun with 20 mice in each cohort, due to the mass euthanasia performed to evaluate amyloid 0484. Overall, in the three visio-spatial memory tests, plaque content (as described in Example 5), Kaplan-Meier mice learned over time, and there was generally improved Survival analysis was performed using the Sub-group of 8 performance in the WT mice as compared to the TG mice, mice in each cohort that were not euthanized. Dosing to the which was expected. There was also a difference between mice in the Sub-groups was continued as described above WT and TG mice in the Elevated Plus Maze. There were through the entirety of the experiment. minimal observed differences in the Rotarod and Open Field Tests, but differences were not expected. Compliance was only a problem in the Barnes Maze, however, when non Example 7 compliant mice were removed the learning trends were present, and the model effect mirrored those seen in the Effect of Intranasal Administration of IgG on MWM and RAWM. Memory 0485 The Morris Water Maze (MWM) Hidden Platform. 0486 MWM is a standard test of spatial memory. MWM 0479. A study was conducted to examine whether intra performance was assessed using hidden-platform testing (4 nasal administration of IgG affects the memory in the brain days, 4 trials/day). Before trials began, the mice were in vivo. The purpose of this study was to examine whether acclimated to Swimming in the water. For each of these blocks of trials, mice were randomly dropped into four chronic treatment with intranasally delivered IgG at two quadrants within the MWM (round tub with water) and doses (0.4 g/kg/2 wk and 0.8 g/kg/2 wk) would have any allowed to swim for 60 seconds or until they reached the effect on memory in a transgenic amyloid mouse model of platform. The mouse's ability to reach the platform Alzheimer's disease. depended on his ability to remember visual cues from Experimental Design: previous trials and their location in relation to the platform. 0480 Mice that did not reach the platform after 60 seconds were 0481. At 15 months of age, the mice described in placed on the platform. Mice were allowed to remain/rest on Example 4 were subjected to a six week battery of behav the platform for 20 seconds between trials. All data was recorded using MouseApp Software, which records escape ioral tests to assess for memory, sensorimotor, and anxiolytic latency. changes. These included Morris water maze hidden and 0487. The Morris Water Maze Visual Platform is visual platform tests (reference memory, visual ability), designed to assess visual ability. It was run just like the radial arm water maze (working memory), passive avoid MWM hidden platform, except the platform was raised just ance task (memory), Barnes maze (memory), open field test above the surface of the water, has a flag on top to identify (exploratory behavior), elevated plus maze (anxiety), and it, and stripes along the side to make it more visual. It was rotarod (motor skills). only run for one day. Analysis was performed the same as 0482 Results: with the MWM hidden platform tests. 0488. Overall, the Morris Water Maze Hidden Platform 0483 For each behavioral test, comparison data was tests showed that there was a clear trend of learning both analyzed using T-tests as described above in Table 23. throughout the week and during individual days, demon Statistical tests were performed on data after removal of strating that the test was effective for measuring memory. both statistical outliers and non-compliant mice, which were Escape latencies were lowest during days 3 and 4, and were specified for each behavioral test. Data was first analyzed by especially lowest during trials 3 and 4 on these days. comparing WT-saline (WT-Sal) mice to TG-saline (TG-Sal) 0489. There was evidence of a transgenic model effect. mice to determine whether there is a transgenic (model) Table 25 and Table 26 show that both WT groups had lower effect for that test. Comparisons between all TG and all WT escape latencies than all three TG groups on days 3 and 4. mice were also performed. Although the latter analysis is WT-Sal mice had lower escape latencies than TG-Sal mice confounded by drug treatment, it gains power by increasing (Table 24, Table 25, Table 26, Table 27, and Table 28). sample size and serves to give an overall picture of a However, when the WT and TG groups were put together, potential transgenic (model) effect. Comparisons were made there were several significant differences, including B1-T2, among individual drug treatment groups. Specifically, the B3-T4, B4-T1, B4-T3, and B4-T4 (p<0.05 or 0.01; Table 24, drug treated TG groups were compared directly to the Table 27, and Table 28). Much of the power for this TG-Saline group to determine whether the drug had any difference came from the TG-high mice, which performed effect. particularly well in this task. US 2017/0044244 A1 Feb. 16, 2017 60

TABLE 24

Summary of T-tests for specific comparisons in behavior tests. Tests are 2-sided and unpaired. Reported numbers are p-values. Gray cells p < 0.05; Boxed cells p < 0.1.

WT-Sal WT-A Test Measure Block Trial vs TG-Sal vs TG-All RAWM Escape ENRA M Escape 2

Escape 4

e W e O.194 WM Escape O.161

WM SC8) 0.446

SC8) 0.959 0.767 O.619 0.65 e 0.069 e 0.995 WM Escape O.281

WM SC8) 0.785

SC8) O.274 O.433 0.627 W Escape ().357 WM Escape O.232 WM Errors 0.538 Errors 2 O.284 0.105 WM MM Errors 3 O.062 O.196 RAWM Errors 4 0.443 0.255 0.577 O.293

O.656 ().753 O.223 O.136 0.227 0.17 0.706 M 0.719 0.385 US 2017/0044244 A1 Feb. 16, 2017 61

TABLE 24-continued

Summary of T-tests for specific com parisons in behavior tests. Tests are 2-sided and unpaired. Reported numbers are p-values. Gray cells p < 0.05; Boxed cells p < 0.1.

O.262 O.186

e O.069 O.086 O.663

e

e e 0.882 0.555 || 0.357 0.702 e

e

e e 1 0.475 0.419 0.906 0.163 e

e e | 4 || 0.355 0.072 0.874 0.6 e

e O.147 0.127 0.264 O.991 Escape

e 0.35 O.696

0.581 O.274 O.826 0.657 O.623 0.052 0.072 O.606 O.138 O.21 O.29 O482 Escape 4 0.925 O.845

O.074 O.282 ().507 O.222 O.863 0.237 O.898 0.448 Open field Line na O.534 O.138 O.112 O.688 US 2017/0044244 A1 Feb. 16, 2017 62

TABLE 24-continued Summary of T-tests for specific comparisons in behavior tests. Tests are 2-sided and unpaired. Reported numbers are p-values. Gray cells p < 0.05; Boxed cells p < 0.1. Crossings

Open field Velocity Elev. plus Time in open arms Elev. plus Frequency in open arms Rotarod Best run na na 0.64 na ().557 Pass. Avoid Escape Learn

TABLE 25 TG-Sal mice had shorter escape latencies than TG-low mice, who performed particularly poor in this task. There was only Average escape latencies (Sec) from the Morris Water Maze tests. one example in which there was a statistical difference Group Day 1 Day 2 Day 3 Day 4 between TG-high and TG-Sal (B1-T1). In this instance, TG-Sal did very well and outperformed the TG-high mice. TG-Low (N = 18) 34.54 30.47 2SO3 24.68 TG-High (N = 18) 33.38 27.06 24.50 30.51 However, it should be noted that the WT-high mice consis TG-Saline (N = 16) 24.80 22.20 24.25 24O2 tently outperformed all other groups in this task. Although WT-High (N = 16) 23.38 23.58 17.73 16.11 T-tests performed at each trial showed no statistical differ WT-Saline (N = 18) 27.26 26.82 24.85 26.82 ences between WT-high and WT-Saline, repeated measures ANOVA would demonstrate a difference between these two groups. TABLE 26 0491 For the MWM hidden platform test, the escape Average escape latencies (sec) from the Morris Water Maze tests latency (time to find the platform) was collected. T-tests with non-compliance removed. were conducted for each day of each trial (1-4). Data was analyzed with non-compliant mice removed in order to more Group Day 1 Day 2 Day 3 Day 4 accurately represent memory. Non-compliant mice were TG-Low (N = 15-18) 31.36 25.53 25.03 23:46 TG-High (N = 15-16) 28.73 20.55 2O.O6 25.57 defined as any mice that had escape latencies of 60 seconds TG-Saline (N = 14-15) 23.25 1968 21.87 1968 (the full time allotted) for trials 3 and 4, when they should WT-High (N = 14-15) 20.93 19.30 14.92 13.18 have been learning to some extent. The percent of non WT-Saline (N = 13-16) 21.65 22.67 18.07 15.87 compliant mice for each group was recorded. For hidden platform tests non-compliance was as follows: TG-low-8. 3%; TG-high-15.3%; TG-saline=7.8%; WT-high-7.8%; TABLE 27 and WT-Saline=18.1%. Average daily escape latencies (Sec) from the Morris Water Maze tests. 0492. The Radial Arm Water Maze (RAWM). Group Day 1 Day 2 Day 3 Day 4 0493 RAWM is used to evaluate short-term, working memory. Similar to a MWM, this test has a round tub with TG ALL (N = 52) 31.14 26.75 24.61 26.50 WT ALL (N = 34) 25.43 25.29 21.50 21.78 water, visual cues throughout the room and a hidden plat form. It is unique in that inserts are placed into the tank to create six radially distributed arms of equal size that ema TABLE 28 nate from the center. Before trials began, the mice were acclimated to Swimming in the water. Mice were dropped Average daily escape latencies (sec) from the Morris Water into 4 radial arms, in an order selected randomly for each Maze tests with non-compliance removed. trial, and given 1 minute to find the platform, with 20 Group Day 1 Day 2 Day 3 Day 4 seconds of rest between each trial. Trials occurred daily for TG ALL (N = 45-49) 27.86 21.92 22.44 22.99 twelve days and each day the platform was moved to a new WT ALL (N = 28-30) 21.29 21.10 16.49 14.43 location. Halfway through the testing, an extra intra-maze visual cue was added to the tank in an effort to make the test 0490. Like with RAWM, the three transgenic groups are a little easier. The visual cue was a large X made of tape grouped closely in Table 25 and Table 26. The only signifi and placed on the inner wall of the maze above the arm with cant differences between TG-Sal and TG-low came on the escape platform. Both errors and escape latency were B1-T1, B2-T2, and B4-T1 (Table 24), and in each case, recorded. US 2017/0044244 A1 Feb. 16, 2017

TABLE 29

RAWM escape latency (seconds) of mice ouped in blocks 1-4.

Block 1 Block 2 Block 3 Block 4 T1(1) T2(1) T3(1) T4(1) T1 (2) T2(2) T3(2) T4(2) T1(3) T2(3) T3(3) T4(3) T1(4) T2(4) T3 (4) T4(4) TG-Low 49.2O SO.48 49.44 45.02 48.06 42.98 40.42 46.66 44.17 35.45 36.25 40.67 37.46 26.44 22.72 29.89 (N = 18) TG-High 49.72 47.13 44.94 47.33 47.78 34.09 42.11 42.96 40.00 45.02 35.56 41.59 32.70 27.20 29.76 29.57 (N = 18) TG-Saline 50.58 44.13 48.96 42.94 49.92 41.58 34.98 43.50 39.94 36.21, 28.45 32.98 32.75 29.06 28.04 29.42 (N = 16) WT-High 39.96 47.11 43.04 42.02 44.81 41.61 34.72 41.70 41.91 33.52 39.54 34.93 40.15 28.28 24.78 24.43 (N = 18) WT-Saline 40.62 38.50 41.76 40.26 43.28 35.98 40.1S 40.81 42.41 31.70 33.83 33.70 33.09 27.87 24.96 25.20 (N = 18)

0494. Overall, RAWM was too difficult for mice in TABLE 32 blocks 1 and 2, as evidenced by a general trend for the escape latency not to go below about 35 seconds (Table 30). RAWM escape latency (seconds) of blocks 1 and 4. After the addition of the extra visual cue in blocks 3 and 4, a clear trend of decreased time to find the platform and errors ESCAPE LATENCY (BLOCK) became apparent in all treatment groups from trial 1 to trial 4 (Table 30, Table 31, and Table 32). This demonstrated that T1(1) T4(1) T1 (2) T4(2) T1(3) T4(3) T1(4) T4(4) the test was effective for measuring memory. TG-Low 49.2O 45.02 48.06 46.66 44.17 40.67 37.46 29.89 TABLE 30 (N = 18) TG-High 49.72 47.33 47.78 42.96 40.00 41.59 32.70 29.57 RAWM escape latency (seconds) of blocks 1 and 2. (N = 18) TG-Saline 50.58 42.94 49.92 43.50 39.94 32.98 32.75 29.42 ESCAPE LATENCY (BLOCK) (N = 16) T1(1) T2(1) T1(2) T2(2) T1(3) T2(3) T1(4) T2(4) WT-High 39.96 42.02 44.81 41.70 41.91 34.93 40.15 24.43 (N = 18) TG-Low 49.2O SO.48 48.06 42.98. 44.17 35.45 37.46 26.44 WT-Saline 40.62 40.26 43.28 40.81 42.41 33.70 33.09 25.20 (N = 18) (N = 18) TG-High 49.72 47.13 47.78 34.09 40.00 45.02 32.70 27.20 (N = 18) TG-Saline 50.58 44.13 49.92 41.58 39.94 36.21 32.75 29.06 0495. There was clear evidence of a transgenic model (N = 16) effect in RAWM (Table 33 and Table 34). In Table 35 an WT-High 39.96 47.11 44.81 41.61 41.91 33.52 40.15 28.28 overall Summary of all groups averaged out over all days (N = 18) shows that in all four trials, both WT groups had lower times WT-Saline 40.62 38.SO 43.28 35.98 42.41 31.70 33.09. 27.87 to find the platform than all three TG groups. This was also (N = 18) true of errors for trials 2-4 (Table 36). In Table 33, Table 34, Table 35, and Table 36, individual blocks and trials can be seen. For escape latency, WT-Sal mice had significantly TABLE 31 shorter escape latencies than TG-Sal mice in B1-T1 (Batch 1-Trial 1), B1-T3, and B3-T2 (p<0.05 or 0.1) (Table 24). For RAWM escape latency (seconds) of blocks 1 and 3. errors (Table 36), WT-Sal mice had significantly fewer errors than TG-Sal mice in B1-T3, B3-T2, and B4-T2 ESCAPE LATENCY (BLOCK (p<0.05 or 0.1) (Table 24). When all WT mice were com T1(1) T3(1) T1(2) T3(2) T1(3) T3(3) T1(4) T3(4) bined and compared to all TG mice (irrespective of treat TG-Low 49.2O 49.44 48.06 40.42 44.17 36.25 37.46 22.72 ment), it was clear that WT mice outperformed TG mice. (N = 18) When all days were combined, WT mice had shorter escape TG-High 49.72 44.94 47.78 42.11 40.00 35.56 32.70 29.76 latency and fewer errors than TG mice in all trials (Table 35 (N = 18) TG-Saline 50.58 48.96 49.92 34.98 39.94 28.45 32.75 28.04 and Table 36). Similarly, in individual blocks and trials, all (N = 16) WT mice had shorter escape latency and fewer errors in all WT-High 39.96 43.04 44.81 34.72 41.91 39.54 40.15 24.78 trials in blocks 2-4 (Table 35 and Table 36). Statistically, WT (N = 18) WT-Saline 40.62 41.76 43.28 40.15 42.41 33.83 33.09 24.96 mice had shorter escape latencies than TG mice in B1-T1, (N = 18) B3-T2, B3-T4, and B4-T4 (p<0.05) (Table 24). Statistically, WT mice had fewer errors than TG mice in B1-T1, B3-T2, B3-T4, B4-T2, and B4-T4 (p<0.05) (Table 24). US 2017/0044244 A1 Feb. 16, 2017 64

TABLE 33 latency and fewer errors than TG mice in all trials (Table 35). Similarly, in individual blocks and trials, all WT mice RAWM escape latencies (seconds) recorded had shorter escape latency and fewer errors in all trials in for 12 days of RAWM testing. blocks 2-4 (Table 35 and Table 36). Statistically, WT mice TG ALL (N = 52 WT ALL (N = 36 had shorter escape latencies than TG mice in B1-T1, B3-T2, B3-T4, and B4-T4 (p<0.05) (Table 24). Statistically, WT Arm Arm Arm Arm Arm Arm Arm Arm mice had fewer errors than TG mice in B1-T1, B3-T2, 1 2 3 4 1 2 3 4 B3-T4, B4-T2, and B4-T4 (p<0.05) (Table 24). Day 1 S1.67 S3.33 49.08 45.24 45.94 45.56 46.81 45.22 0497. The Barnes Maze. Day 2 S1.1S 45.58 48.54 46.87 36.11 40.22 40.06 37.47 Day 3 46.6O 43.19 45.6O 43.41 38.86 42.64 40.33 40.72 0498. The Barnes maze is a visual memory task based on Day 4 SO.29 37.31 39.87 43.27 43.17 37.67 37.03 34.83 finding an escape hole on a table, aided by visual cues Day 5 49.85 40.62 38.27 44.76 41.94 38.14 36.61 44.08 throughout the room. The table was round, elevated 1 m Day 6 45.41 40.45 39.84 45.20 47.OO 40.58 38.67 44.86 from the floor, and had 40 escape holes spaced equally Day 7 45.76 38.76 37.14 42.38 41.53 32.08 34.53 40.67 Day 8 38.79 41.61 39.20 38.92 43.36 28.36 38.36 28.97 around the periphery of the table. One of these holes had an Day 9 39.7S 36.79 24.71 34.63 41.57 37.39 37.17 33.31 escape box directly underneath, while the others were open. Day 10 34.42 29.90 29.69 29.94 39.81 27.19 27.50 25.47 The motivation to find the escape box was aversive stimuli Day 11 34.13 23.69 24.10 31.15 35.50 27.50 26.08. 28.94 in the form of bright lights and fans blowing above the Day 12 34.54 28.94 26.60 27.81 34.56 29.53 21.03 20.03 surface of the table. The escape box was located in one location for the duration of the study. The mouse was given 4 days, with 3 trials/day to learn the location of the escape TABLE 34 RAWM escape latencies (seconds) of blocks 1-4. Block 1 Block 2 Block 3 Block 4 T1(1) T2(1) T3(1) T4(1) T1 (2) T2(2) T3(2) T4(2) T1(3) T2(3) T3(3) T4(3) T1(4) T2(4) T3 (4) T4(4) TG ALL 49.81 47.37 47.73 45.18 48.54 39.45 39.32 44.40 41.42 39.04 33.62 38.62. 34.37 27.51. 26.79 29.63 (N = 52) WT ALL 40.29 42.81 42.40 41.14 44.06 38.80 37.44 41.26 42.16 32.61 36.69 34.31 36.62. 28.07 24.87 24.81 (N = 36)

TABLE 35 box. Mice were given up to two minutes on the table to find the escape hole. If after 2 minutes they did not find the 12 day average of RAWM escape latencies (seconds). escape box, they were placed into the box. Both escape latency to find the hole and errors were recorded and Trial 1 Trial 2 Trial 3 Trial 4 analyzed. Errors were defined as head-pokes through holes TG ALL (N = 52) 43.53 38.35 36.89 39.46 that do not have the escape box. WT ALL (N = 36) 40.78 35.57 35.35 35.38 0499. Overall, the Barnes maze test did not work well for the mice in this study. This was the only behavior test in which non-compliance was an issue (roughly 50% of all TABLE 36 mice did not perform the task). While running the tests, the mice were generally not scared of the aversive stimuli. 12 day average of RAWM errors (trial averages). However, among the mice that were compliant and included in the analyses, there was a learning trend across the days Trial 1 Trial 2 Trial 3 Trial 4 and trials, which can be seen in the escape latencies. TG ALL (N = 52) 4.77 4.64 4.28 4.42 0500. There was evidence of a model effect with this test. WT ALL (N = 36) 4.52 3.84 3.71 3.72 Table 37 and Table 38 shows that both WT groups have lower escape latencies on days 3 and 4 than all three TG groups. This mirrors data collected with the RAWM and 0496 There was evidence of a TG model effect in MWM tests, the other two long-term memory tasks. This RAWM. A summary of all groups averaged out over all days difference is also seen when all WT mice and TG mice were (Table 35) shows that in all four trials, both WT groups had combined as in Table 39 and Table 40. lower times to find the platform than all three TG groups. This was also true of errors for trials 2-4 (Table 35 and Table TABLE 37 36). In Table 35 and Table 36, individual blocks and trials can be seen. For escape latency, WT-Sal mice had signifi Average escape latencies (Sec) from the Barnes Water Maze by treatment. cantly shorter escape latencies than TG-Sal mice in B1-T1 (Batch 1-Trial 1), B1-T3, and B3-T2 (p<0.05 or 0.1) (Table Time (s) Day 1 Day 2 Day 3 Day 4 24). As shown in Table 36, WT-Sal mice had significantly TG-Low (N = 18) 105.15 99.76 95.44 85.67 fewer errors than TG-Sal mice in B1-T3, B3-T2, and B4-T2 TG-High (N = 18) 107.74 94.57 100.30 97.33 TG-Saline (N = 16) 95.48 89.10 90.10 82.31 (p<0.05 or 0.1) (Table 24). When all WT mice were com WT-High (N = 17) 99.06 95.98 93.65 82.04 bined and compared to all TG mice (irrespective of treat WT-Saline (N = 18) 94.15 97.41 93.43 87.63 ment), it was clear that WT mice outperformed TG mice. When all days were combined, WT mice had shorter escape US 2017/0044244 A1 Feb. 16, 2017 65

TABLE 38 TABLE 42 Average escape latencies (sec) from the Barnes Water Average errors from the Barnes Water Maze by genotype. Maze by treatment with non-compliance removed. Time (s) Day 1 Day 2 Day 3 Day 4 Day 1 Day 2 Day 3 Day 4 TG-Low (N = 7-12) 82.81 78.25 83.60 72.75 TG ALL (N = 52) 8.97 6.31 5.53 S.O1 TG-High (N = 6-8) 84.28 72.14 75.71 68.86 WT ALL (N = 35) 8.25 7.48 S.O3 4.46 TG-Saline (N = 7-9) 79.71 65.38 74.30 71.48 WT-High (N = 8-10) 83.74 79.17 66.29 60.13 WT-Saline (N = 7-10) 69.17 68.43 73.00 60.74 0502. For Barnes Maze, both the escape latency (time to find the escape hole) and errors (number of times a mouse pokes his head into a hole that does not have the escape box) TABLE 39 were collected. T-tests were conducted for each day of each Average escape latencies (sec) from the Barnes Water Maze by genotype. trial (1-3). Data was analyzed with non-compliant mice removed in order to more accurately represent memory. Time (s) Day 1 Day 2 Day 3 Day 4 Non-compliant mice were defined as any mice that had TG ALL (N = 52) 103.07 94.69 95.48 88.67 escape latencies of 120 seconds (the full time allotted) for WT ALL (N = 35) 96.53 96.71 93.53 84.91 trials 3, when they should have been learning to some extent. The percent of non-compliant mice for each group was recorded and was as follows: TG-low-48.6%; TG-high-61. TABLE 40 1%; TG-saline=48.4%; WT-high 45.6%; and WT-sa line=52.8%. Average escape latencies (sec) from the Barnes Water Maze by genotype with non-compliance removed. 0503 Elevated Plus Maze. Time (s) Day 1 Day 2 Day 3 Day 4 0504. The Elevated Plus Maze is a standard test of TG All (N = 21-28) 82.05 72.21 78.16 71.37 baseline anxiety in which the animal is placed in the center WT All (N = 17-19) 76.88 74.75 70.02 60.42 of an elevated 4-arm maze that consists of two arms that are open and two arms that are enclosed. The number of times 0501. There was no evidence of a drug effect in the the animal entered each of the arms and the time spent in Barnes Maze tests (Table 37, Table 38, Table 39, Table 40, each arm over 4 minutes was recorded. The test was used to Table 41, Table 42). The only statistical significance was in determine the unconditioned response to a potentially dan B4-T1, in which TG-low mice performed very poorly and gerous environment (the open, unprotected arms) and anxi had longer escape latency than TG-Sal mice (p<0.1: Table ety-related behavior was measured by the degree to which 24). the rodent avoids the open arms of the maze. (0505. There was a transgenic effect in the Elevated Plus TABLE 41 Maze. In this model, all TG mice spent more time and made Average number of errors from the Barnes Water Maze by treatment. more frequent arm entries into the open arms of the maze than all WT mice, demonstrating inhibition of exploratory Day 1 Day 2 Day 3 Day 4 behavior and anxiety that WT mice have regarding open TG-Low (N = 18) 8.48 5.57 6.24 5.39 spaces. When WT-Sal mice were compared to TG-Sal mice, TG-High (N = 18) 6.85 S.S4 4.15 4.04 TG mice spent significantly more time and have signifi TG-Saline (N = 16) 11.90 8.02 6.29 S.69 WT-High (N = 17) 6.96 6.45 4.69 4.OO cantly more arm entries into the open arms (Table 24, Table WT-Saline (N = 18) 9.46 8.44 5.35 4.89 43, and Table 44). When all WT-mice and all TG-mice were combined, the same results were seen (Table 44 and Table 45), p<0.05: Table 24). TABLE 43 Average time spent in open arms during the Elevated Plus Maze.

TIME (SEC

SUM PERCENTAGE

Avg. Time Avg Time Std Error Std Error Avg. Time Avg Time Std Error Std Error Enclosed Open Enclosed Open Enclosed Open Enclosed Open

TG-Low (N = 18) 115.2 31.4 10.8 5.3 48.0 13.1 4.5 2.2 TG-High (N = 16) 128.7 48.9 11.2 8.7 53.7 20.4 4.7 3.6 TG-Saline (N = 117.5 34.6 11.6 7.4 49.0 14.4 4.9 3.1 15) WT-High (N = 151.9 20.6 8.6 3.7 63.4 8.6 3.6 1.6 16) US 2017/0044244 A1 Feb 16, 2017 66

TABLE 43-continued Average time spent in open arms during the Elevated Plus Maze.

TIME (SEC

SUM PERCENTAGE Avg. Time Avg Time Std Error Std Error Avg. Time Avg Time Std Error Std Error Enclosed Open Enclosed Open Enclosed Open Enclosed Open WT-Saline (N = 1698 15.9 11.6 4.4 70.8 6.6 4.8 1.8 16) TG ALL (N = 49) 120.3 38.1 6.4 4.2 SO.2 15.9 2.7 1.8 WT ALL (N = 32) 1608 18.3 7.3 2.9 67.1 7.6 3.0 1.2

TABLE 44 Average frequency of entries into open arms during the Elevated Plus Maze. FREQUENCY SUM PERCENTAGE Avg. Freq Avg. Freq Std Error Std Error Avg. Freq Avg. Freq Std Error Std Error Enclosed Open Enclosed Open Enclosed Open Enclosed Open TG-Low (N = 18) 16.6 10.8 2.O 2.0 60.9 39.1 S.1 S.1 TG-High (N = 16) 16.O 14.8 2.2 3.6 57.8 42.2 5.2 5.2 TG-Saline (N = 15) 15.9 8.1 2.4 2.2 69.6 3O4 5.8 5.8 WT-High (N = 16) 13.8 3.4 1.5 O.S 82.1 17.9 2.4 2.4 WT-Saline (N = 16) 9.8 3.0 1.1 O.8 81.9 18.1 3.8 3.8 TG ALL (N = 49) 16.2 11.3 1.2 1.6 62.5 37.5 3.1 3.1 WT ALL (N = 32) 11.8 3.2 1.O 0.5 82.O 18.0 2.2 2.2

0506. There was no evidence of a drug effect in the TABLE 45 Elevated Plus Maze tests. Although the TG-high group had the most arm-entries and spent the most time in the open Average velocity of mice. arms, it was not significantly different from any other groups (Table 24, Table 43, and Table 44). Avg. Velocity Std Dev Stol Error 0507 For the Elevated Plus Maze, both the time spent in i. N. 8. open and enclosed arms and the number of arm entries (also TG Saline (N = 15) 8.73 2.78 0.72 called frequency of arm entries) were recorded. Mice were WT High (N = 16) 10.03 2.50 O.63 not included in the analyses if they fell off the maze in less WT Saline (N = 17) 9.71 3.38 O.82 than 120 seconds. There were 3 mice that fell off, all from different groups. For outliers, mice were removed if both their time spent in open arms and frequency of entries into TABLE 46 open arms were more than two standard deviations from the Average Velocity of mice, averaged by genotype. mean of their treatment group. Outliers included 3 mice, all from different groups. Avg. Velocity Std Error 0508. The Open Field Maze Test. WLNS s: 0509. The Open Field Maze Test is used to detect any change in spontaneous locomotor activity due to drug treat ment or anxiety. Each mouse was given 4 minutes to TABLE 47 individually explore a rectangular box, while being tracked by the EthoVision video tracking system. For analysis, the Average number of line Crossings by mice. box was Subdivided into 16 equally sized squares that are Avg Line separated by manually drawn lines using the “line draw” Crossings Stod Dev Std Error feature in EthoVision. The number of line crossings and TG Low (N = 18) 87.56 32.93 7.76 patterns of exploration were measured. TG High (N = 18) 110.94 44.01 10.37 0510. There was no evidence of a transgenic or drug WiNS 325, 2. effect in the Open Field Maze tests. All groups of mice had WT Saline (N = 17) 112.59 33.44 8.11 very similar line crossings and velocity (Table 24, Table 45, Table 46, Table 47, and Table 48). US 2017/0044244 A1 Feb. 16, 2017 67

TABLE 48 TABLE 51-continued Average number of line crossings by mice, averaged by genotype. Trial averages of run time (Sec) on the rotarod by treatment group. Avg Line Crossings Std Error Trial 1 Trial 2 Trial 3 TG ALL (N = 51) 102.65 5.33 TG-Saline (N = 16) 16.60 25.63 24.60 WT ALL (N = 33) 107.83 6.21 WT-High (N = 16) 19.56 43.00 44.2O WT-Saline (N = 18) 17.00 17.06 32.39 TG ALL (N = 52) 15.72 20.44 33.72 0511 For the Open Field Maze, both the number of line WT ALL (N = 34) 16.60 25.63 24.60 crossings and the overall velocity were measured. Outliers were removed if an individual mouse's line crossings were more than 2 standard deviations from the mean of the 0515. There was no evidence of a drug effect among treatment group. This included 3 mice, each from different transgenic groups (Table 49, Table 50, and Table 51). treatment groups. Analysis was performed for both line However, it was observed that the WT-high mice had longer crossings and Velocity. times on the rotarod than the WT-Sal mice. A t-test between 0512. The Rotarod Performance Test. WT-Sal and WT-high yielded a p-value of 0.089 for the 0513. The Rotarod Performance Testis used to detect any longest run, and a p-value of 0.041 for the average run changes in endurance, balance, and coordination. Mice were (T-tests not shown, Table 49, Table 50, and Table 51). placed on an automated rotating bar and allowed to walk on 0516 For the Rotarod test, the time on the rotating bar the bar for up to 60 seconds. The speed of rotation was before the mouse fell off was recorded. Three trials were gradually increased and the rodent’s ability to remain on the conducted. If a mouse reached 120 seconds (the maximum rotating bar was recorded as the total time spent on the bar. time) before trial 3, subsequent runs were not conducted. For Mice were given three trials, and the best time is used for each treatment group, both the average time on the bar and analysis. the maximum time on the bar for each mouse were analyzed. 0514. There was no transgenic model effect on the Data could not be recorded if the mouse did not stay on the Rotarod tests. All groups performed essentially the same and rod long enough before starting (~3 seconds), and there was there were no statistical differences among groups (Table 24 only 1 mouse that did not stay on long enough to start for all and Table 49, Table 50, and Table 51). There was a non three trials. significant trend for all WT mice to outperform all TG mice 0517 The Passive Avoidance Task. (Table 49, Table 50, and Table 51). 0518. The Passive Avoidance Task is a classical condi tioning test used to assess short-term or long-term memory TABLE 49 for mice and rats. The passive avoidance apparatus consists of equal-sized light and dark compartments with a lightbulb Longest average runs on the rotarod by treatment group. fixed in the center of the roof of the light compartment. The Best Trial (Average) (sec) floor consists of a metal grid connected to a shocker. The two compartments are separated by a trap door. On the learning TG-Low (N = 18) 30.59 TG-High (N = 18) 37.33 day (day 1), a mouse was placed in the light compartment TG-Saline (N = 16) 35.38 and the time taken to enter the dark compartment was WT-High (N = 16) S4.13 recorded and termed as initial latency. Immediately after the WT-Saline (N = 18) 35.67 mouse entered the dark chamber a door was automatically TG ALL (N = 52) 37.33 closed and an electric footshock (0.7 mA) was delivered for WT ALL (N = 34) 35.38 3 seconds. Twenty-four hours after the acquisition trial, a second retention trial was conducted and the time the mouse takes to enter the dark compartment as designated retention TABLE 50 latency (RL: recorded to a maximum of 500 seconds, no shock is administered during this entry). T-tests were per Average run time on the rotarod by treatment group. formed to compare the effects of IN IgG WT vs. TG. Avg. Time (Sec) 0519. Whereas RAWM, MWM hidden platform, and TG-Low (N = 18) 1943 Barnes maze tests all showed evidence of learning and TG-High (N = 18) 23.30 improved learning in WT mice over TG mice, this test TG-Saline (N = 16) 22.35 consistently showed the opposite effect, regardless of drug WT-High (N = 16) 35.24 treatment. There was no evidence of a drug effect among WT-Saline (N = 18) 22.25 TG ALL (N = 52) 23.30 transgenic groups (Table 24, Table 52, Table 53, and Table WT ALL (N = 34) 22.35 54). TABLE 52 TABLE 51 Passive avoidance learn day escape latency (Sec). Trial averages of run time (Sec) on the rotarod by treatment group. Learn Esc. St. Err Trial 1 Trial 2 Trial 3 TG-Low (N = 17) 445 9.4 TG-High (N = 19) 46.3 7.6 TG-Low (N = 18) 1O.S3 21.25 27.06 TG-Saline (N = 15) 43.7 9.2 TG-High (N = 18) 15.72 20.44 33.72 WT-High (N = 17) 21.6 6.4 US 2017/0044244 A1 Feb. 16, 2017 68

TABLE 52-continued among groups (Table 24). The one statistical difference came in trial 1, due to a strong performance by WT-Sal that Passive avoidance learn day escape latency (Sec). did not carry over into Subsequent trials. There was also no Learn Esc. St. Err evidence of a drug effect among transgenic groups (Table 24, Table 55, Table 56, Table 57, and Table 58). However, WT-Saline (N = 18) 22.4 3.9 TG ALL (N = 51) 44.9 4.9 much like with the MWM hidden platform tests, there was WT ALL (N = 35) 22 3.6 a trend for WT-high mice to outperform all other groups (Table 55, Table 56, Table 57, and Table 58). T-test com parisons between WT-Sal and WT-high for each individual trial were not significant, but a T-test for all trials between TABLE 53 these two groups had a p-value of 0.06. Passive avoidance test day escape latency (Sec). TABLE 55 Test Esc. St. Err Visual escape (Sec) by treatment group. TG-Low (N = 15) 224.6 8.5 TG-High (N = 17) 229.5 8.3 Group Trial 1 Trial 2 Trial 3 Trial 4 Average TG-Saline (N = 13) 2O7.0 16.8 WT-High (N = 16) 114.3 22.O TG-Low (N = 18) 34.83 33.44 39.17 30.22 33.44 WT-Saline (N = 18) 153.8 20.9 TG-High (N = 18) 31.44 33.67 35.33 37.89 33.67 TG ALL (N = 45) 2214 4.9 TG-Saline (N = 16) 23.19 36.56 29.75 28.94 36.56 WT ALL (N = 34) 135.2 3.6 WT-High (N = 16) 28.25 23.44 23.13 22.00 23.44 WT-Saline (N = 18) 29.78 29.06 26.11 25.50 29.06

TABLE 54 TABLE 56 Passive avoidance average of escape latency differences (see). Visual escape (Sec) by treatment group, with non-compliance removed. Average of Differences St. Err Group Trial 1 Trial 2 Trial 3 Trial 4 Average TG-Low (N = 15) 190.2 8.5 TG-Low (N = 13) 3O46 29.15 31.15 18.77 29.15 TG-High (N = 17) 1919 8.2 TG-High (N = 13) 21.69 27.92 25.85 29.38 27.92 TG-Saline (N = 13) 175.1 16.8 TG-Saline (N = 14) 17.93 33.21 25.43 24.50 33.21 WT-High (N = 16) 98.8 22.4 WT-High (N = 14) 25.07 18.57 17.86 16.57 18.57 WT-Saline (N = 18) 131.4 21.4 WT-Saline (N = 17) 3141 27.24 24.12 23.47 27.24 TG ALL (N = 45) 186.5 6.3 WT ALL (N = 34) 116.1 15.5 TABLE 57 0520. This test demonstrated an unexpected TG effect. Whereas TG mice with impaired memory should normally Visual escape (Sec) by genotype group. have trouble remembering not to enter the dark chamber and receive a shock after training, this was not the case. TG mice Group Trial 1 Trial 2 Trial 3 Trial 4 Average generally did not enter the chamber on the test day, whereas TG ALL (N = 52) 30.08 34.48 34.94 32.48 34.48 WT mice seemed not to care whether they received a shock WT ALL (N = 34) 29.06 26.41 24.71 23.85 26.41 on the test day. These results can be seen in Table 52, Table 53, and Table 54. The poor performance of the WT mice compared to the TG mice is statistically significant (p<0.05; TABLE 58 Table 24). The same willingness for WT mice to enter the dark chamber can be seen in the learning phase and may play Visual escape (Sec) by genotype, with non-compliance removed. a role in the willingness of normal, WT mice to go receive a painful shock. Group Trial 1 Trial 2 Trial 3 Trial 4 Average 0521 For the Passive Avoidance Task, the escape latency TG ALL (N = 40) 23.23 30.18 27.43 24.23 30.18 on both the learning day (day 1) and the test day (day 2) were WT ALL (N = 31) 28.55 23.32. 21.29 20.35 23.32 recorded and the difference between the escape latency between the test and learn day were calculated. Mice were 0524 For the visual platform MWM, the escape latency not run on the test day (day 2) if they did not receive a shock (time to find the platform) was collected. T-tests were on day 1, which included 7 mice spread across 4 groups. conducted for each day of each trial (1-4). Data was ana Mice did not receive a shock simply because they did not lyzed with non-compliant mice removed in order to more enter the dark chamber. There were no outliers calculated. accurately represent memory. Non-compliant mice were Analyses were performed for the learn trial and the test trial. defined as any mice that had escape latencies of 60 seconds 0522 Morris Water Maze Visual Platform. (the full time allotted) for trials 3 and 4, when they should 0523. Differences in performance in this test were not have been learning to some extent. The percent of non expected as all mice were genetically tested for the RD1 compliant mice for each group was recorded. For visual gene and the mice did not have problems with vision. There platform tests non-compliance was as follows: TG-low-6. was no transgenic model effect. All groups performed essen 9%; TG-high-6.9%; TG-saline=3.1%; WT-high 3.1%; and tially the same and there were no statistical differences WT-Saline=1.4%. US 2017/0044244 A1 Feb. 16, 2017 69

Example 8 0529 Adult male Sprague Dawley rats (N=6, average weight 250 g) and adult male C57blk mice (n-6, 7-8 weeks) Radiolabeled 'I IgG Reaches the CNS with were used for Phase 1. Adult male Sprague Dawley rats Intranasal Delivery (N=18, average weight 264 g) were used for Phase 2. The 0525. A study was conducted to determine the feasibility animals were housed in pairs with free access to food and and to optimize the methods used to determine the amount water and were kept on a 12 h light cycle. of intravenously and intranasally delivered radiolabed "I 0530 Prior to commencing the Phase 1 and 2 experi IgG reaching the CNS in rats and mice at a two hour time ments, the animals were allowed to normalize in the facility point. for a period of three days before handling occurred. Animals 0526 Experimental Design: were slowly acclimated to human handling over a period of 0527 There were two phases of this experiment. In phase about two weeks. Enrichment food treats are given after 1, six mice and rats were used to test a variety of different handling to encourage a human-animal bond while the methods including anesthesia with 2 hour Survival, drug acclimation process proceeds. Restraint techniques were administration methods (intravenous infusion through can kept brief and facilitated by using a towel, restraint device, nulations of the jugular vein in rats and mice, intranasal tube or Scruffing, when working with mice. method in rats), transcardial perfusion (with and without a 0531. An anesthesia cocktail containing Ketamine HCl non-ionic detergent), and tissue processing for capillary (30 mg/kg), Xylazine HCl (6 mg/kg), and Acepromazine (1 depletion and gamma counting. Animals and the methods mg/kg) was used. All anesthesia was administered as Sub tested with each are shown in Table 51. cutaneous injections. Boosters alternated between the Cock TABLE 59 Experimental design of phase 1 of Example 8. R = rat and M = mouse. Brain Animal Surgery IV delivery IN delivery Perfusion Dissection a-R-1 Jugular Vein No infusion IN tube Saline Who e Brain Cannulation method COval a-R-2 Jugular Vein 2 mg/mL BSA IN tube 0.05% Triton X Who e Brain Cannulation until death method COval a-R-3 Jugular Vein 2 mg/mL BSA IN tube Saline Capi lary Cannulation over 1 hour method Depletion a-R-4 Jugular Vein No infusion No IN delivery 0.1% Triton X Who e Brain Cannulation COval a-R-5 Jugular Vein No infusion IN tube 0.1% Triton X Who e Brain Cannulation method COval a-R-6 Jugular Vein No infusion No IN delivery Saline Capi lary Cannulation Depletion a-M-1 Jugular Vein No infusion No IN delivery Saline Capi lary Cannulation Depletion a-M-2 Jugular Vein 2 mg/mL BSA No IN delivery 0.05% Triton X Who e Brain Cannulation over 1 hour COval a-M-3 Jugular Vein 2 mg/mL BSA No IN delivery 0.1% Triton X Who e Brain Cannulation over 1 hour COval a-M-4 Jugular Vein 2 mg/mL BSA No IN delivery Saline Who e Brain Cannulation over 1 hour COval a-M-5 Jugular Vein 8 g/kg IgG No IN delivery 0.05% Triton X Who e Brain Cannulation over 1 hour COval a-M-6 Jugular Vein 8 g/kg IgG No IN delivery 0.05% Triton X Who e Brain Cannulation over 1 hour COval

0528. In Phase 2, three tissue processing techniques after tail above and 50 mg/kg Ketamine. Reflexes were tested to administration of high IVIG does in 18 rats were tested in assess level of anesthesia every 10-15 minutes throughout order to determine the optimal technique of Subsequent the study. Phase 1 experiments. The 18 rats were divided into 3 0532. Intranasal deliver in rats was performed using a experimental groups (Table 60). specialized pipette tip. The specialized pipette tip was inserted into the rat naris. The pipette tip was created by TABLE 60 cutting 23 mm off the end of a gel loading pipette tip and attaching a 16 mm length of tubing (ID=0.04 mm, OD=0.07 Experimental groups for Phase 2. mm). The tubing was placed over the wide end of the pipette Group 1 Group 2 Group 3 tip with an overlap of 5.5 mm, and a black mark with a sharpie was made at 14.5 mm from the narrowest end of the 2I-IVIG dose 200 mg 200 mg 200 mg pipette tip. The narrow end was ultimately inserted into the Perfusion 140 mL saline 140 mL saline 90 mL saline, 25 mL. with capillary 0.025% Triton X-100, rat’s nose up to the black mark. depletion 25 mL saline 0533. For intranasal delivery, the fully anesthetized rat = 6 rats 6 rats 6 rats was placed on its back on a heating pad in a metal Surgical tray. The heating pad and rectal probe was used to maintain the rat’s core temperature at 37° C. A 2"x2" gauze pad was US 2017/0044244 A1 Feb. 16, 2017 70 rolled into a pillow and was securely taped. The pillow was sections (see FIG. 5). Each section was hemisected and then placed under the rat’s neck to ensure that the underside placed into tubes for counting. from nostril to torso was planar and horizontal. 0537 For capillary deletion, each brain section was 0534. A lead impregnated shield was placed between the weighed and transferred to an ice cold ground glass homog Surgical tray and the experimenter for protection against onizer. A volume of 2.857 times the tissue sample weight of buffer, pH 7.4 (10 mM HEPES, 141 mM NaCl, 4 mM KC1, radiation. The dose solution, pipette, pipette tips, and waste 2.8 mM CaCl2, 1 mM MgSO. H.O. 1 mM NaH2PO, and receptacle were arranged behind the shield for easy access. 10 mM D-Glucose), was added to the homogonizer. The The modified pipette tip was inserted into the rat naris up to brain sample was homogenized using vertical strokes. A the black mark. The sample to be delivered (40-50 ul) was small volume of 26% dextran solution was added to the drawn into a pipettor, the tip of the pipettor placed into the homogenized brain sample in order to provide a final open tube at the end of the modified pipette tip (while concentration of 15.5% Dextran in the homogenate. The carefully holding the modified pipette tip in place in the rats homogenate was then Vortexted, homogenized for a second nose), and then the entire dose was slowly expelled into the time with vertical strokes, and then decanted into a small rat’s nostril. glass centrifuge tube. The homogenate was then centrifuged 0535. After the animals were euthanized, their brains in a Swinging bucket rotor for 15 minutes at 4°C. at a speed were removed for analysis. With a large Surgical Scissors, the of 5400xg. The homogenate was separated into the follow head of the animal was removed by cutting dorsal to ventral ing layers: a bottom pellet containing the capillary segments, to avoid contamination. Using a scalpel, the fur and skin on a clear liquid layer, and a top "cream layer containing the the top of the skull was cut from nose to point of decapi nervous tissue. Using a transfer pipette, the cream and clear tation. The skin was folded back and held with a small gauze liquid layers were transferred into new tubes. The radioac pad to expose the top of the skull. Using a small hemostat, tivity of the Supernatant and the pellet was determined using the remainder of the spinal column was chipped away a gamma counter. exposing the upper cervical spinal cord and posterior brain 0538 Results: (cerebellum). Next, the top of the skull was removed to the 0539. The data from Phase 2 shows that intravenous olfactory bulbs exposing the entire dorsal side of the brain. 'I-IVIG reached the central nervous system. The animals The hemostat was inserted with one blade scraping the with capillary depletion tissue processing had the most IVIG ventral surface of the skull. This ensured the integrity of the in the brain tissue (49,791 ng). The animals perfused with dorsal Surface of the brain was maintained. A Small spatula 0.025% Triton X as a second perfusate had the least IVIG in was used to loosen the lateral surfaces of the brain from the the brain tissue (33.855 ng) (Table 61 and Table 62). The skull and dura. The brain was inverted over a clean Petri capillary depletion pellet which should hold all of the IVIG dish. The optic nerve was severed, which released the brain stuck to the capillary walls only accounted for -3% of the from the skull. The brain was assessed for quality of whole brain IVIG in those animals (Table 63). The low perfusion. amount of IVIG in the capillary pellet could be a result of 0536 The brain was placed dorsal side up. A single edged homogenization friction during processing, releasing the razor blade was used to sever the olfactory bulbs from the IVIG stuck to the capillary walls and allowing it to be mixed brain at the natural angle. Olfactory bulbs were collected. in with the Supernatant instead of staying with the capillaries Razor blades were used to cut the brain into seven coronal in the pellet. TABLE 61

- 2I-IVIGIV SPSSS present inII theSSSI central nervousSEWOS systemSYSS measuredSSIS in CPM.SM Perfusate Total CPM Total CPM Total CPM Total CPM Total CPM CPMI

Rat Method Whole Brain Liquid Pellet R. Hemisphere L. Hemisphere (2nd) (3rd) b-1 Cap Dep 68,554 65,326 3,228 30,687 37,867 b-4 Cap Dep 40,791 39,372 1419 28,352 2,439 b-7 Cap Dep 29,048 28,229 819 13,374 5,674 b-10 Cap Dep 15,498 14,851 647 8,104 7,393 b-13 Cap Dep 47,908 46,533 1,376 28,757 9,151 b-16 Cap Dep 69,964 68,128 1,836 29.458 40,505 b-3 Control 98.341 52,972 45,368 278 356 b-6 Control 21,141 10,557 0,584 112 144 b-11 Control 36,457 19,077 7,380 141 121 b-15 Control 28,303 14,228 4,075 126 66 b-17 Control 20,524 9,508 1,016 231 127 b-18 Control 38,683 19,350 9,333 125 73 b-2 Triton X 36,984 16,622 20,362 S4O 216 b-5 Triton X 49,882 25,617 24,264 98 219 b-8 Triton X 19,194 11,031 8,163 243 no sample b-9 Triton X 33,716 15,026 8,690 422 82 b-12 Triton X 21,255 7,639 3,616 527 151 b-14 Triton X 14,013 6,712 7,301 441 117 Average Cap Cep 45,294 43,740 1,554 23,122 22,172 Average Control 40,575 20,949 9,626 169 148 Average Triton X 29,174 13,775 5,399 379 157 US 2017/0044244 A1 Feb. 16, 2017 71

TABLE 62 ng by Group

Perfusate Total ng Total ng Total ng Total ng Total ng ngul Rat Method Whole Brain Liquid Pellet R. Hemisphere L. Hemisphere (2nd) (3rd) b-1 Cap Dep 68,537 65,310 3,227 30,679 37,858 b-4 Cap Dep 45,383 43,804. 1579 31,544 13,840 b-7 Cap Dep 32,060 31,156 904 4,761 17,300 b-10 Cap Dep 18,231 17470 761 9,534 8,697 b-13 Cap Dep 57,258 55,614 1,644 34,369 22,889 b-16 Cap Dep 77.276 75,248 2,028 32,537 44,739 b-3 Control 108.404 58,393 50,011 306 392 b-6 Control 24,824 2,397 12,428 132 169 b-11 Control 35,411 8,530 16,881 137 118 b-15 Control 36,686 8,442 18,244 163 86 b-17 Control 25,940 2,017 13,923 292 160 b-18 Control 50,757 25,390 25,367 16S 95 b-2 Triton X 46,547 20,921 25,626 68O 272 b-5 Triton X 56,294 28,910 27,383 111 247 b-8 Triton X 22,577 2,975 9,601 285 no sample b-9 Triton X 39,032 7,396 21,637 488 95 b-12 Triton X 22,099 7,943 14,157 S48 157 b-14 Triton X 16,581 7.942 8,639 522 138 Average Cap Dep 49,791 48,101 1,690 25,571 24.220 Average Control 47,004 24,195 22,809 199 170 Average Triton X 33,855 16,014 17,841 439 182

TABLE 63 ng by Group

Percent Percent of Percent of est. of ng Percent Percent of ng ng est. ng in est. ng in Percent delivered of whole whole delivered delivered est. ng in 2nd 3rd ng in (Whole brain brain (2nd (3rd blood perfusate perfusate blood Brain) (Liquid) (Pellet) perfusate) perfusate) Average 124,564,379 62% O.02% 97% 3% Cap Dep Average 151,853,766 4978.470 4.249,634 76% O.02% 2.5% 2.1% Control Average 134,662,521 10,980,039 4,543,372 67% O.02% 5.5% 2.3% Triton X *The total estimated blood volume was determined as the body weight times 0.06 plus 0.77 (Lee and Blaufox, 1985).

(0540. The Triton X perfusion methods resulted in a 28% 0541. In these results, approximately 0.02% of the total reduction of IVIG whole brain concentration versus the delivered IVIG that was infused reached the brain (Table 55) saline perfusion control. The perfusate should show the in all methods. During the Phase 2 experiments it was noted amount of IVIG cleared from the blood vessels over the that the brain tissues were slightly pinkish, Suggesting the course of the 25 ml (perfused at a rate of 15 ml/min). Three total Volume perfused was not adequate to completely 250 ul samples of each perfusate were counted in the gamma remove blood from the brain. This slight coloration appeared counter. Averages of the three were than calculated. To consistent throughout all animals in each experimental determine the total amount of IVIG in each perfusate, the group. An increase in the total volume of perfusate in the ng/ul IVIG concentration was determined and multiplied by next Phase should solve this issue. 25000 (the 25 ml of perfusate used). The first perfusates (-90 ml at 15 ml/min) were not collected since this step was the same in all of the animals in the study. In the group Example 9 perfused with 0.025% Triton X, more ''I-IVIG was removed (439 ng/ul) than the groups perfused with saline Biodistribution of IgG Administered Intranasally (199ng/ul). This difference was not seen in the 3" perfusate, and Intravenously in Mice meant to clear any remaining Triton X from the blood vessels, (170 ng/ul and 182 ng/ul, respectively) (Tables 1 0542. A study was conducted to compare the biodistri and 2) Suggesting that the maximum clearance of IVIG from bution of pooled human immunoglobulin G (IgG) adminis the vessels at this concentration of Triton X was achieved. tered to mice intranasally and intravenously. Delivery of IgG A higher Triton X concentration in the perfusate may yield to the brain and residual IgG in the bloodstream were a further reduction. determined.