Compositions for neutralizing liver-expressed antimicrobial peptide 2

Abstract

Provided herein are anti-LEAP2 aptamers, that specifically binds liver expressed antimicrobial peptide 2 (LEAP2, LEAP-2) protein, or a derivative thereof and pharmaceutical composition comprising one or more of these aptamers. Provided herein are methods of using the aptamers and / or compositions thereof to treat various diseases and disorders linked to growth hormone secretagogue receptor (GHSR). Also, provided herein are combination therapies combining the aptamers and / or compositions thereof, with anti-LEAP2 shRNA and / or GHSR-agonists.

Description

Atty. Docket Number: UTSDP3820WO- 1001380566TITLE COMPOSITIONS FOR NEUTRALIZING LIVER-EXPRESSED ANTIMICROBIAL PEPTIDE 2CROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims priority to U.S. Provisional Patent Application Serial No.63 / 763,130, filed February 25, 2025, and titled “COMPOSITIONS FOR NEUTRALIZING LIVER-EXPRESSED ANTIMICROBIAL PEPTIDE 2,” which is incorporated by reference herein in its entirety.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted in ST26 format and is hereby incorporated by reference in its entirety. Said ST26 copy, created on February 24, 2026, is named UTSD.P3820WO_SEQ_LIST.xml and is 9,609 bytes in size.BACKGROUND1. Field

[0003] The present disclosure encompasses compositions for reducing or neutralizing liver-expressed antimicrobial peptide 2 (LEAP 2) and methods for treating metabolic diseases using the disclosed compositions.2. Discussion of Related Art

[0004] Metabolic dysregulation plays a critical role in a variety of conditions, impacting energy balance, muscle function, and overall physiological homeostasis. Chronic illnesses such as cancer, chronic obstructive pulmonary disease (COPD), chronic kidney disease, and heart failure can lead to significant metabolic alterations, including muscle wasting and changes in adipose tissue composition. Unlike simple starvation, these conditions often involve systemic inflammation, hormonal imbalances, and impaired nutrient utilization, which contribute to adverse clinical outcomes, including reduced physical function, poor quality of life, and increased mortality.

[0005] The prevalence and severity of metabolic disorders vary depending on the underlying disease. For example, obesity affects a significant portion of the global population and is a major risk factor for cardiovascular disease, diabetes, and fatty liver disease. In contrast, muscle wasting syndromes such as sarcopenia and cachexia are more commonly observed in aging populations and individuals with chronic illnesses. Cachexia, characterized by severe muscle wasting and weight loss, is frequently associated with advanced cancer (affecting 50-80% of patients), end-stage heart failure (5-15%), and chronic kidney disease. These301871278Atty. Docket Number: UTSDP3820WO- 1001380566metabolic conditions, whether linked to excessive weight gain, insulin resistance, or severe muscle atrophy, significantly impact morbidity and mortality.

[0006] Ghrelin, a peptide hormone primarily produced in the stomach, plays a crucial role in regulating appetite, energy homeostasis, and metabolic function. It binds to the growth hormone secretagogue receptor (GHSR) (also known as the ghrelin receptor), a G-protein-coupled receptor (GPCR) expressed in several brain regions, the pituitary gland, pancreatic islets, gastrointestinal tract, and other tissues. Activation of GHSR stimulates appetite, promotes fat storage, and modulates anabolic and catabolic pathways, thereby influencing body weight, muscle maintenance, and overall energy balance. Beyond its effects on metabolism, ghrelin is neuroprotective and regulates gastrointestinal motility, muscle function, learning and memory, mood, anxiety, muscle strength, and growth hormone release, among other physiological processes.

[0007] Therapeutic strategies targeting GHSR have shown promise in a range of metabolic disorders. Given the widespread impact of metabolic dysregulation across various conditions, targeting the ghrelin-GHSR axis presents a promising approach for developing new treatments that address energy balance, muscle maintenance, and metabolic health.SUMMARY

[0008] In some aspects, the current disclosure encompasses an anti-LEAP2 aptamer, that specifically binds the liver expressed antimicrobial peptide 2 (LEAP2, LEAP-2) protein, or a derivative thereof. In some aspects, the aptamer comprises a nucleic acid sequence. In some aspects, the nucleic acid sequence is a deoxyribonucleic acid, or a variant thereof. In some aspects, the nucleic acid sequence comprises a sequence as set forth in SEQ ID NO: 2 [GCTACGTG], or a sequence at least about 95%, 96%, 97%, 98%, 99%, or 100% identical thereto. In some aspects, the nucleic acid sequence comprises a sequence as set forth in SEQ ID NOs: 3-6, or a sequence at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical thereto. In some aspects, the nucleic acid sequence comprises a sequence as set forth in SEQ ID NOs: 7-10, or a sequence at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical thereto. In some aspects, the aptamer further comprises one or more modifications. In some aspects, the one or more modifications comprise nucleobase modifications, sugar modifications, backbone modifications, end modifications, or any combination thereof. Non-limiting examples of nucleobase modifications include incorporation of 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil,301871278 - 2 -Atty. Docket Number: UTSDP3820WO- 1001380566dihydrouracil, beta-D-galactosylqueosine, inosine, N6-sopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methyl cytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid, 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, 2,6-diaminopurine, biotinylated analogs, 2'-Fluoro (2'-F) modification, 2'-O-Methyl (2'-OMe) modification, thiol groups, amine groups, and fluorescent labels, and any combinations thereof. Non-limiting examples of sugar modifications include incorporation of 2'-Fluoro (2'-F), 2'-O-Methyl (2'-OMe), 2’-O-Methoxyethyl (2’-MOE), 2’-O-Amino (2’-NH2), arabinose, 2-fluoroarabinose, xylose, and hexose, and any combinations thereof. Non-limiting examples of backbone modifications include incorporation of phosphorothioate, a phosphorodithioate, a phosphoramidothioate, a phosphoramidate, a phosphordiamidate, a methylphosphonate, an alkyl phosphotriester, formacetal PEGylation, circularization, spacer additions, multimerization, locked nucleic acids (LNA), unlocked nucleic acids, xeno nucleic acids, peptide nucleic acids (PNA), and end caps, and any combinations thereof. In some aspects, the one or more modifications comprise 5’-biotinylation, 5’-PEGylation, 3’-PEGylation, or any combination thereof. In some aspects, the current disclosure also encompasses a polynucleotide comprising one or more anti-LEAP2 aptamers disclosed herein.

[0009] Also disclosed herein are compositions comprising one or more aptamer, that specifically bind the liver expressed antimicrobial peptide 2 (LEAP2, LEAP-2) protein, or a derivative thereof. In some aspects, the composition may further comprise one or more anti-LEAP2 shRNAs, and / or one or more GHSR agonists. In some aspects, the composition is a pharmaceutical composition, comprising at least one pharmaceutically acceptable excipient. In some aspects, the administration of the pharmaceutical composition to a subject in need thereof reduces synthesis, levels of LEAP2 protein, and / or LEAP2 binding with growth hormone secretagogue receptor (GHSR) in the subject. In some aspects, the subject has, or is suspected of having any one or more of cachexia (cancer cachexia, cardiac cachexia, pulmonary cachexia, renal cachexia, cachexia of aging), anorexia nervosa, hypoglycemia, depression, anxiety, Alzheimer’s disease, gastrointestinal dysmotility disorders (postoperative ileus, diabetic gastroparesis), or short stature. In some aspects, the pharmaceutical composition reduces ghrelin resistance and / or enhances the activity of endogenous ghrelin, or ghrelin-independent GHSR activity, and / or increases the activity of administered ghrelin, and / or increases the activity of an administered GHSR agonist in the subject. In some aspects,301871278 - 3 -Atty. Docket Number: UTSDP3820WO- 1001380566the composition comprises a nanoparticle, a conjugate particle, a viral vector, a hydrogel, a liposome, an exosome, or a cell. In some aspects, the composition is for intravenous, intracranial, intrathecal, subcutaneous, intranasal, cranial, transmucosal, trans-nasal, transcranial, intracerebroventricular, intestinal, and / or parenteral delivery.

[0010] In some aspects, the current disclosure also encompasses a method of treating a disease or a disorder, in a subject in need thereof, the method comprising administering to the subject one or more anti-LEAP2 aptamers, alone or in combination with one or more anti-LEAP2 shRNAs, and / or one or more GHSR agonists, or any combination thereof. In some aspects, the subject has, or is suspected of having a ghrelin, or GHSR linked disease or disorder. In some aspects, the ghrelin, or GHSR linked disease or disorder is a cachexia. In some aspects, the cachexia is a cancer cachexia, cardiac cachexia, pulmonary cachexia, renal cachexia, or cachexia of aging. In some aspects, the ghrelin or GHSR linked disease or disorder is anorexia nervosa, bulimia nervosa, hypoglycemia, depression, anxiety, Alzheimer’s disease, dementia, age related memory decline, gastrointestinal dysmotility disorders (postoperative ileus, diabetic gastroparesis), hypoglycemia unawareness, post-traumatic stress disorder, amyotrophic lateral sclerosis or other neurodegenerative disorders, type 1 diabetes, type 2 diabetes, obesity, short stature, or Prader Willi Syndrome. In some aspects, the method comprises administering to the subject the one or more anti-LEAP2 aptamers, and the one or more anti-LEAP2 shRNAs. In some aspects, the one or more anti-LEAP2 aptamers, and the one or more anti-LEAP2 shRNAs are administered concurrently or separately. In some aspects, the method further comprises administering one or more GHSR agonists, increasing the activity of administered ghrelin, or increasing the activity of administered GHSR agonist, or any combination thereof. In some aspects, GHSR agonist is administered concurrently or separately with the administration of the one or more anti-LEAP2 aptamers and / or the one or more anti-LEAP2 shRNAs.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. Aspects of the present disclosure may be better understood by reference to one or more of these drawings in combination with the detailed description of specific aspects presented herein.

[0012] FIGS. 1A-1D provide anti-LEAP2 DNA aptamer characterization by GHSR-Tango -arrestin recruitment assay. FIG. 1A shows the effect of the 5’-biotinylated forms of anti-LEAP2 DNA aptamers 2, 3, 7, and 8 on LEAP2-induced GHSR (Ghrelin receptor) deactivation were examined using HTI_A cells expressing GHSR-Tango plasmid. One-way ANOVA analysis. * P < 0.01 compared to 10 nM LEAP2 + ghrelin group, t P < 0.001 compared to10 nM LEAP2301871278 - 4 -Atty. Docket Number: UTSDP3820WO- 1001380566+ ghrelin group, n = 16 for the 10 nM ghrelin only and 10 nM ghrelin + 10 nM LEAP2 groups; n = 8 for all other groups. FIG. 1B shows the effect of 4 different modified versions of anti-LEAP2 DNA aptamer 7 (5’ biotin, 5’ PEG, 3’ PEG, and 5’ + 3’ PEG modifications) on LEAP2-induced GHSR deactivation were examined using HTI_A cells expressing GHSR-Tango plasmid. One-way ANOVA analysis, t P < 0.001 compared to 10 nM LEAP2 + ghrelin group, n = 16 for the 10 nM ghrelin only and 10 nM ghrelin + 10 nM LEAP2 groups; n = 8 for all the other groups. FIG. 1C shows the ECso of anti-LEAP2 aptamer 7 with 5’ biotin modification was determined (1.26 nM) using HTLA cells expressing GHSR-Tango plasmid. Nonlinear regression model of log(agonist) vs response to calculate ECso. Each dose was a mean of 8 independent replicates ± SEM. FIG. 1D provides quantitative determination of LEAP2 ICso without aptamer and with 5’-biotinylated aptamer 7 in HTLA cells expressing GHSR-Tango plasmid. Nonlinear regression model of log(antagonist) vs response to calculate ICso- Each dose was a mean of 8 independent replicates ± SEM. Competitive enzyme kinetics equation was used to determine the Ki and Vmax. Curves were fitted using GRAPHPAD PRISM™ 9.0.

[0013] FIG. 2 shows a standard curve for the quantitative PCR assay designed / performed to detect administered biotinylated anti-LEAP2 aptamer 7 in plasma of mice (Aptamer detection assay).

[0014] FIG. 3 is a bar graph showing detected LEAP2 aptamer levels in the blood samples from mice post administration using intravenous (IV), intraperitoneal (IP), or subcutaneous (SC) modes of administration.

[0015] FIG. 4 is a bar graph showing that animals treated with LEAP2 aptamer exhibit greater stimulation (P=0.088) in ghrelin-induced growth hormone (GH) release. All data are presented as mean± s.e.m. Two-way ANOVA followed by Tukey’s comparisons test were used for GH assay to test for significant differences among test groups.

[0016] FIGS. 5A-5B show ghrelin-induced food intake after 1 hour and 2 hours post ghrelin injection, and the effect of LEAP2 aptamer pre-administration on ghrelin-induced food intake. Food was reintroduced immediately after ghrelin injection. A single pre-weighed standard chow pellet was placed on the cage floor, and the amount consumed during the following 1-hour (FIG. 5A) and 2-hour (FIG. 5B) periods was measured. LEAP2 aptamer enhanced ghrelin-induced food intake at both timepoints.

[0017] The drawing figures do not limit the present disclosure to the specific aspects disclosed and described herein. The drawings are not necessarily to scale, emphasis instead being placed on clearly illustrating principles of certain aspects of the present disclosure.301871278 - 5 -Atty. Docket Number: UTSDP3820WO- 1001380566DETAILED DESCRIPTION

[0018] The following detailed description references the accompanying drawings that illustrate various aspects of the present disclosure. The drawings and description are intended to describe aspects of the present disclosure in sufficient detail to enable those skilled in the art to practice the present disclosure. Other components can be utilized, and changes can be made without departing from the scope of the present disclosure. The following description is, therefore, not to be taken in a limiting sense.

[0019] LEAP2 (Liver-expressed antimicrobial peptide 2) is an endogenous peptide hormone, produced mainly by the liver and small intestine. It plays a crucial role in metabolic regulation by acting as an antagonist and inverse agonist of the growth hormone secretagogue receptor (GHSR or ghrelin receptor), thereby inhibiting ghrelin-mediated action and reducing food intake. LEAP2 counterbalances the effects of ghrelin, a hormone that impacts appetite, gastrointestinal motility, muscle function, energy homeostasis, learning and memory, mood, anxiety, muscle strength, and growth hormone release, among other physiological processes.

[0020] Administration of LEAP2 and analogs of LEAP2 have been shown to reduce food intake, reduce body weight, and reduce metabolic dysfunction-associated steatotic liver disease. Additionally, LEAP2 has been implicated in immune function and exhibits antimicrobial properties, contributing to innate immunity. Conversely, deletion of the LEAP2 gene in mice enhances sensitivity to administered ghrelin, resulting in increased food intake and elevated growth hormone secretion. This heightened response underscores LEAP2's critical role in modulating GHSR activity and maintaining energy balance.

[0021] In some aspects, the current disclosure is based on the observation that LEAP2 limits the actions of the hormone ghrelin to stimulate food intake, increase body weight, raise blood glucose (or prevent falls in blood glucose), act as an antidepressant and anxiolytic, increase survival, enhance memory, increase muscle function, stimulate gastrointestinal motility, and stimulate GH secretion. Interestingly, during these studies, it was found that aptamers that specifically target LEAP2 can enhance food intake in mice administered with these aptamers. In some aspects, and without being bound by theory, these LEAP2 aptamers act like neutralizing antibody that reduces plasma LEAP2 levels. By reducing LEAP2, these aptamers can be effective in reducing the inhibition of GHSR, thereby treating GHSR linked disease or disorder. The disclosed aptamers may be combined with other therapeutics that adversely impact the levels of LEAP2 directly or indirectly, and / or enhance ghrelin receptor signaling. Thus, the aptamers, or pharmaceutical compositions comprising these aptamers may be used in-combination therapeutics with anti-LEAP2, or GHSR agonist, or both.301871278 - 6 -Atty. Docket Number: UTSDP3820WO- 1001380566I. Terminology

[0022] The phraseology and terminology employed herein are for the purpose of description and should not be regarded as limiting. For example, the use of a singular term, such as, “a” is not intended as limiting of the number of items. Also, the use of relational terms such as, but not limited to, “top,” “bottom,” “left,” “right,” “upper,” “lower,” “down,” “up,” and “side,” are used in the description for clarity in specific reference to the figures and are not intended to limit the scope of the present disclosure or the appended claims.

[0023] Any term of degree such as, but not limited to, “substantially” as used in the description and the appended claims, should be understood to include an exact, or a similar, but not exact configuration. For example, “a substantially planar surface” means having an exact planar surface or a similar, but not exact planar surface.

[0024] The terms “comprising,” “including,” and “having” are used interchangeably in this disclosure. The terms “comprising,” “including,” and “having” mean to include, but not necessarily be limited to the things so described.

[0025] The terms “or” and “and / or,” as used herein, are to be interpreted as inclusive or meaning any one or any combination. Therefore, “A, B, or C” or “A, B, and / or C” mean any of the following: “A,” “B,” or “C”; “A and B”; “A and C”; “B and C”; “A, B, and C.” An exception to this definition will occur only when a combination of elements, functions, steps, or acts are in some way inherently mutually exclusive.

[0026] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this disclosure belongs. The following references provide one of skill with a general definition of many of the terms used in this disclosure: Singleton et al., Dictionary of Microbiology and Molecular Biology (3rd ed. 2006); Chambers Dictionary of Science and Technology(Walker ed., 1999); The Glossary of Genetics, 5th Ed., R. Rieger et al. (2008), The Harper Collins Dictionary of Biology (1991), all of which are incorporated by reference herein. As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise.

[0027] The phraseology and terminology employed herein are for the purpose of description and should not be regarded as limiting. When introducing elements of the present disclosure or the preferred aspects(s) thereof, the articles “a,” “an,” “the,” and “said” are intended to mean that there are one or more of the elements. The terms “comprising,” “including,” and “having” are intended to be inclusive and mean that there may be additional elements other than the listed elements. Wherever the terms “comprising” or “including” are used, it should be understood the disclosure also expressly contemplates and encompasses additional aspects301871278 - 7 -Atty. Docket Number: UTSDP3820WO- 1001380566“consisting of” the disclosed elements, in which additional elements other than the listed elements are not included.

[0028] The term “about” or “approximately,” as used herein, can mean within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation, per the practice in the given value. Where particular values are described in the application and claims, unless otherwise stated the term “about” can mean an acceptable error range for the particular value, such as 10% of the value modified by the term “about.” As used herein, the term “about,” can mean relative to the recited value, e.g., amount, dose, temperature, time, percentage, etc., ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%.

[0029] As used herein, the term ‘between’ refers to a range that includes the two values recited as the limits of the range, as well as all values greater than or equal to the lower limit and less than or equal to the upper limit, unless expressly stated otherwise. For example, ‘between 10% and 50%’ includes 10%, 50%, and all integer and non-integer values therein.

[0030] As used herein, “treatment,” “therapy,” and / or “therapy regimen” refer to the clinical intervention made in response to a disease, disorder or physiological condition manifested by a patient or to which a patient may be susceptible. The aim of treatment includes the alleviation or prevention of symptoms, slowing or stopping the progression or worsening of a disease, disorder, or condition and / or the remission of the disease, disorder or condition.

[0031] As used herein, “prevent” or “prevention” refers to eliminating or delaying the onset of a particular disease, disorder or physiological condition, or to the reduction of the degree of severity of a particular disease, disorder or physiological condition, relative to the time and / or degree of onset or severity in the absence of intervention.

[0032] As used herein, “individual,” “subject,” “host,” and “patient” can be used interchangeably herein and refer to any mammalian subject for whom diagnosis, treatment, prophylaxis or therapy is desired, for example, humans, pets, livestock, horses or other animals. As used herein, the term “subject” and “patient” are used interchangeably herein and refer to both human and nonhuman animals. The term “nonhuman animals” of the disclosure includes all vertebrates, e.g., mammals and non-mammals, such as nonhuman primates, sheep, dog, cat, horse, cow, chickens, amphibians, reptiles, and the like. In some aspects, the subject can be a human. In other aspects, the subject can be a human in need of treating a cancer.301871278 - 8 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0033] As used herein, the term “aptamer” refers to a single-stranded nucleic acid molecule, including DNA, RNA, or modified nucleic acids, that folds into a three-dimensional structure capable of specifically binding to liver-expressed antimicrobial peptide 2 (LEAP2) or a derivative thereof. Aptamers contemplated herein may include chemically modified nucleotides, modified sugars, modified backbones, terminal modifications (e.g., biotinylation or PEGylation), or any combination thereof, and may be isolated or optimized using SELEX or analogous enrichment methods.II. LEAP2 aptamer

[0034] Disclosed herein are anti-LEAP2 aptamers, that specifically bind the liver expressed antimicrobial peptide 2 (LEAP2, LEAP-2) protein, or a derivative thereof. Typically, nucleic acid aptamers are single-stranded nucleic acid (DNA or RNA) ligands that function by folding into a specific globular structure that dictates binding to target proteins, nucleic acids, or other molecules with high affinity and specificity. Aptamers are usually isolated from libraries consisting of 1014-1015random oligonucleotide sequences by a procedure termed SELEX (systematic evolution of ligands by exponential enrichment) as further described in the Examples.

[0035] In some aspects, the aptamers may be oligomers of artificial ssDNA, RNA, XNA (Xeno nucleic acids), or peptides that bind a region on the LEAP2 peptide. In some aspects, the aptamer may bind two or more contiguous amino acids of the LEAP2 polypeptide sequence, wherein the LEAP2 polypeptide sequence comprises an amino acid sequence as set forth in SEQ ID NO: 1 (Uniprot ID: Q969E1).SEQ ID NO: 1:MWHLKLCAVLMIFLLLLGQIDGSPIPEVSSAKRRPRRMTPFWRGVSLRPIGASCRDDSECIT RLCRKRRCSLSVAQE

[0036] In some aspects, the aptamer may bind two or more structurally contiguous amino acids of the LEAP2 polypeptide sequence. In some aspects, an anti-LEAP2 aptamer may reduce the levels of, or inhibit LEAP2 protein function. In some aspects, the anti-LEAP2 aptamer is a DNA aptamer, RNA aptamer, a protein aptamer, or a variant thereof.

[0037] In some aspects, a disclosed aptamer is a DNA aptamer. In some aspects, the DNA aptamer comprises a sequence as set forth in SEQ ID NO: 2 (SEQ ID NO: 2: GCTACGTG). or a sequence at least 95%, 96%, 97%, 98%, 99%, or 100%, identical thereto. In some aspects, the DNA aptamer comprises a nucleic acid sequence as set forth in any one of SEQ ID NOs: 3-10 (Table 1), or a sequence at least about 80% identical thereto. In some aspects, the DNA aptamer comprises a nucleic acid sequence as set forth in any one of SEQ ID NOs:301871278 - 9 -Atty. Docket Number: UTSDP3820WO- 10013805662-10 (Table 1), or a sequence at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, identical thereto.Table 1: Aptamer core and extended sequencesIII. Modification of Nucleic Acids

[0038] In certain aspects, the nucleic acid of the disclosure may be modified. A “modified nucleic acid” refers to a molecule in which one or more of the components of the nucleic acid, namely sugars, bases, and phosphate moieties, are different from that which occur in nature, for example, different from that which occurs in the human body. Several modifications to nucleic acids are described in the art. These modifications may improve properties such as resistance to nucleases, permeability across biological membranes, solubility, stability, or modulation of pharmacokinetic and pharmacodynamics properties while maintaining specificity to the target mRNA. For example, the modifications on the nucleotides can include, but are not limited to, LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-C-allyl, 2'-301871278 - 10 -Atty. Docket Number: UTSDP3820WO- 1001380566fluoro, 2'-deoxy, 2'-hydroxyl, and combinations thereof. It is contemplated that one or more of these modifications may be excluded in an aspect.Modified Bases

[0039] Therapeutic nucleic acid may include natural (i.e. A, G, II, C, or T) or modified (e.g. 7-deazaguanosine, inosine, etc.) bases. Modification of bases includes the incorporation of modified bases (or modified nucleoside or modified nucleotides) that are variations of standard bases, sugars and / or phosphate backbone chemical structures occurring in ribonucleic (i.e., A, C, G and II) and deoxyribonucleic (i.e., A, C, G and T) acids. Included or excluded within this scope are, for example: Gm (2'-methoxyguanylic acid), Am (2'-methoxyadenylic acid), Cf (2'-fluorocytidylic acid), Ilf (2'-fluorouridylic acid), Ar (riboadenylic acid). The aptamers may also include cytosine or any cytosine-related base including 5-methylcytosine, 4-acetylcytosine, 3-methylcytosine, 5-hydroxymethyl cytosine, 2-thiocytosine, 5-halocytosine (e.g., 5-fluorocytosine, 5-bromocytosine, 5-chlorocytosine, and 5-iodocytosine), 5-propynyl cytosine, 6-azocytosine, 5-trifluoromethylcytosine, N4,N4-ethanocytosine, phenoxazine cytidine, phenothiazine cytidine, carbazole cytidine or pyridoindole cytidine. The aptamer may further include guanine or any guanine-related base including 6-methylguanine, 1-methylguanine, 2,2-dimethylguanine, 2-methylguanine, 7-methylguanine, 2-propylguanine, 6-propylguanine, 8-haloguanine (e.g., 8-fluoroguanine, 8-bromoguanine, 8-chloroguanine, and 8-iodoguanine), 8-aminoguanine, 8-sulfhydrylguanine, 8-thioalkylguanine, 8-hydroxylguanine, 7-methylguanine, 8-azaguanine, 7-deazaguanine or 3-deazaguanine. The aptamer may still further include adenine or any adenine-related base including 6-methyladenine, N6-isopentenyladenine, N6-methyladenine, 1 -methyladenine, 2-methyladenine, 2-methylthio-N6-isopentenyladenine, 8-haloadenine (e.g., 8-fluoroadenine, 8-bromoadenine, 8-chloroadenine, and 8-iodoadenine), 8-aminoadenine, 8-sulfhydryladenine, 8-thioalkyladenine, 8-hydroxyladenine, 7-methyladenine, 2-haloadenine (e.g., 2-fluoroadenine, 2-bromoadenine, 2-chloroadenine, and 2-iodoadenine), 2-aminoadenine, 8-azaadenine, 7-deazaadenine or 3-deazaadenine. Also included are uracil or any uracil-related base including 5-halouracil (e.g., 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil), 5-(carboxyhydroxylmethyl)uracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, dihydrouracil, 1-methylpseudouracil, 5-methoxyaminomethyl-2-thiouracil, 5'-methoxycarbonylmethyluracil, 5-methoxyuracil, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid, pseudouracil, 5-methyl-2-thiouracil, 2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl)uracil, 5-methylaminomethyluracil, 5-propynyl uracil, 6-azouracil, or 4-thiouracil.301871278 - 11 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0040] The nucleic acids of the disclosure may include or exclude modified base variants. The modified base variant may include or exclude 4-acetylcytidine, 5-(carboxyhydroxylmethyl)uridine, 2'-methoxycytidine, 5-carboxymethylaminomethyl-2-thioridine, 5-carboxymethylaminomethyluridine, dihydrouridine, 2'-O-methylpseudouridine, b-D-galactosylqueosine, inosine, N6-isopentenyladenosine, 1 -methyladenosine, 1-methylpseudouridine, 1-methylguanosine, 1-methylinosine, 2,2-dimethylguanosine, 2-methyladenosine, 2-methylguanosine, 3-methylcytidine, 5-methylcytidine, N6-methyladenosine, 7-methylguanosine, 5-methylaminomethyluridine, 5-methoxyaminomethyl-2-thiouridine, b-D-mannosylqueosine, 5-methoxycarbonylmethyluridine, 5-methoxyuridine, 2-methylthio-N6-isopentenyladenosine, N-((9-b-D-ribofuranosyl-2-methylthiopurine-6-yl)carbamoyl)threonine, N-((9-b-D-ribofuranosylpurine-6-yl)N-methyl-carbamoyl)threonine, u rd ine- 5- oxy acetic acid methylester, uridine-5-oxyacetic acid (v), wybutoxosine, pseudouridine, queosine, 2-thiocytidine, 5-methyl-2-thiouridine, 2-thiouridine, 4-thiouridine, 5-methyluridine, N-((9-b-D-ribofuranosylpurine-6-yl)carbamoyl)threonine, 2'-O-methyl-5-methyluridine, 2'-O-methyluridine, and wybutosine, and 3-(3-amino-3-carboxypropyl)uridine.

[0041] Also included are the modified nucleobases described in U.S. Pat. Nos. 3,687,808, 3,687,808, 4,845,205, 5,130,302, 5,134,066, 5,175,273, 5,367,066, 5,432,272, 5,457,187, 5,459,255, 5,484,908, 5,502,177, 5,525,711, 5,552,540, 5,587,469, 5,594,121, 5,596,091, 5,614,617, 5,645,985, 5,830,653, 5,763,588, 6,005,096, and 5,681,941, each of which is incorporated herein by reference in its entirety.Modified Sugars

[0042] Modified sugar moieties for use in nucleic acids are well known in the art and are described for example in U.S. Pat. No. 9,045,754 which is incorporated by reference herein in its entirety. Modified sugars can be used to alter, typically increase, the affinity of the nucleic acid for its target and / or increase nuclease resistance. For example, in some aspects, the binding affinity of the nucleic acids to their target can be increased by incorporating substituent groups in the nucleoside subunits of the nucleic acids. In some aspects, the substituent groups are T substituent groups, substituent groups located at the 2' position of the pentofuranosyl sugar moieties of the nucleoside subunits of the nucleic acids. Substituent groups include or exclude fluoro, alkoxy, amino-alkoxy, allyloxy, imidazolylalkoxy and polyethylene glycol. Alkoxy and aminoalkoxy groups generally include lower alkyl groups, particularly C1-C9 alkyl. In a particular aspect, the 2' substituent group is 2'-O-methyl. Polyethylene glycols are of the structure (O — CH2 — CH2)n — O-alkyl. In a particular aspect, the substituent is a polyethylene glycol substituent of the formula ( — O — CH2 — CH2)n — O-alkyl, wherein n=1 and alkyl=CH3. This modification has been shown to increase both affinity of an oligonucleotide for its target301871278 - 12 -Atty. Docket Number: UTSDP3820WO- 1001380566and nuclease resistance of an oligonucleotide. See U.S. Pat. No. 7,629,321 cited above. A further particularly useful 2'-substituent group for increasing the binding affinity is the 2'-fluoro group.

[0043] Examples of modified nucleoside and nucleotide sugar backbone variants known in the art include, without limitation, those having, e.g., 2' ribosyl substituents such as F, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2, CH3, ONO2, NO2, N3, NH2, OCH2CH2OCH3, O(CH2)2ON(CH3)2, OCH2OCH2N(CH3)2, O(C1-10 alkyl), O(C2-10 alkenyl), O(C2-10 alkynyl), S(C1-10 alkyl), S(C2-10 alkenyl), S(C2-10 alkynyl), NH(C1-10 alkyl), NH(C2-10 alkenyl), NH(C2-10 alkynyl), and O-alkyl-O-alkyl. Desirable 2' ribosyl substituents include 2'-methoxy (2-OCH3), 2'-aminopropoxy (2' OCH2CH2CH2NH2), 2-0-allyl (2-CH2— CH=CH2), 2'-O-allyl (2'-O— CH2— CH=CH2), 2'-amino (2'-NH2), and 2'-fluoro (2'-F). The 2'-substituent may be in the arabino (up) position or ribo (down) position. One or more of these variants may be excluded from aspects of the disclosure.

[0044] Another class of modified nucleic acids known in the art and that may be utilized in the nucleic acids of the disclosure contain alkyl modifications at the 2' position of the ribose moiety. These nucleic acids were developed to improve the binding affinity and hybridization stability with target mRNA, and to increase the nuclease resistance of the nucleic acids. In this category, the most commonly used nucleic acids are 2'-O-Methyl (2-OME) and 2-0-Methoxyethyl (2-MOE) nucleic acids. Nucleic acids with this type of modification are incapable of activating RNAse H. Therefore, to induce RNAse H activation, chimeric nucleic acids have been developed in which a central gap region consisting of a phosphorothioate deoxyribose core is flanked with nuclease resistant arms such as 2-OME or 2-MOE that possess greater nuclease resistance. A “gapmer” is produced as a result, in which RNAse H can sit in the central gap and activate target specific mRNA degradation, while the arms prevent the nucleic acid degradation. Nucleic acids in this category may possess higher affinity for mRNA, show better tissue uptake, and have increased resistance to nucleases, longer in vivo half-life, and lesser toxicity, as compared to the modified nucleic acids of the first class.

[0045] A further class of nucleic acids known in the art and that may be utilized in the nucleic acids of the disclosure contain modifications of the furanose ring along with modifications of the phosphate linkage, the ribose moiety, or the nucleotides. These modifications were designed to improve the nuclease stability, target affinity and pharmacokinetic profiles of the nucleic acids. Common examples of third category of nucleic acids are Locked nucleic acid (LNA), Peptide nucleic acid (PNA) and Morpholino phosphoroamidates (MF) nucleic acids in this category are more stable in biological fluids because of their high resistance to degradation by nucleases and peptidases. They also exhibit a strong hybridization affinity with301871278 - 13 -Atty. Docket Number: UTSDP3820WO- 1001380566the mRNA. Further, PNAs recognize double stranded DNA, and are able to modulate gene expression or induce mutation by strand invasion of chromosomal duplex DNA. Nucleic acids in this category also do not activate RNAse H and rely on sterically hindering the ribosomal machinery to cause translational arrest. They do not bind to serum proteins as they are uncharged. Lack of charge reduces the odds of non-specific interactions but increases the rate of clearance from the body. Their electrostatically neutral backbones may reduce solubility and make uptake more difficult.

[0046] A representative list of preferred modified sugars includes but is not limited to bicyclic modified sugars (BNA’s), including methyleneoxy (4-CH2 — O-2') BNA and ethyleneoxy (4-(CH2)2 — O-2' bridge) BNA; substituted sugars, especially 2'-substituted sugars having a 2'-F, 2-OCH3 or a 2'-O(CH2)2 — OCH3 substituent group; and 4'-thio modified sugars. Sugars can also be replaced with sugar mimetic groups among others. Methods for the preparations of modified sugars are well known to those skilled in the art.Modified Internucleotide Linkages

[0047] Nucleic acid therapeutics may further comprise at least one phosphorothioate or methylphosphonate internucleotide linkage. The phosphorothioate or methylphosphonate internucleotide linkage modification may occur on any nucleotide of the sense strand or antisense strand or both (in nucleic acid therapeutics including a sense strand) in any position of the strand. For instance, the internucleotide linkage modification may occur on every nucleotide on the sense strand or antisense strand; each internucleotide linkage modification may occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand may contain both internucleotide linkage modifications in an alternating pattern. The alternating pattern of the internucleotide linkage modification on the sense strand may be the same or different from the antisense strand, and the alternating pattern of the internucleotide linkage modification on the sense strand may have a shift relative to the alternating pattern of the internucleotide linkage modification on the antisense strand.

[0048] In certain aspects, the nucleic acids of the disclosure comprise or exclude one or more nucleoside subunits connected by phosphorus linkages including phosphodiester, phosphorothioate, 3'(or -5')deoxy-3'-(or -5')thio-phosphorothioate, phosphorodithioate, phosphoroselenates, 3'-(or -5')deoxy phosphinates, borano phosphates, 3'-(or 5'-)amino phosphoramidates, hydrogen phosphonates, borano phosphate esters, phosphoramidates, alkyl or aryl phosphonates and phosphotriester phosphorus linkages. In some aspects, the nucleic acids of the disclosure comprise nucleoside subunits connected by carbonate, carbamate, silyl, sulfur, sulfonate, sulfonamide, formacetal, thioformacetyl, oxime,301871278 - 14 -Atty. Docket Number: UTSDP3820WO- 1001380566methyleneimino, methylenemethylimino, methylenehydrazo, methylenedimethylhydrazo and methyleneoxymethylimino linkages.

[0049] For example, one class of modified nucleic acids described in the art and that may be utilized in the nucleic acids of the disclosure are those that have one of the non-bridging oxygen atoms in the phosphate group of the nucleic acids replaced with either a sulfur group (phosphorothioates), a methyl group (methyl phosphonates) or an amine group (phosphoramidates). These nucleic acids have greater resistance to nucleases and longer plasma half-life as compared with phosphodiester oligonucleotides. They are capable of activating RNAse H, carry negative charges which facilitate their delivery to cells, and have suitable pharmacokinetics. Among these modifications, phosphorothioate modifications are used most widely. For example, Vitravene, an FDA approved ASO drug, and most of the other ASO drugs in clinical trials are phosphorothioate ASOs.

[0050] In addition, the bases in nucleotide may be joined by a linkage other than a phosphodiester bond, so long as it does not interfere with hybridization. Thus, inhibitory nucleic acids may be peptide nucleic acids in which the constituent bases are joined by peptide bonds rather than phosphodiester linkages. The inhibitory nucleic acids may be prepared by converting the RNA to cDNA using known methods (see, e.g., Ausubel et. al., Current Protocols in Molecular Biology Wiley 1999). The inhibitory nucleic acids can also be cRNA (see, e.g., Park et. al., (2004) Biochem. Biophys. Res. Commun. 325(4): 1346-52).Covalent linkers

[0051] Conjugate moieties are attached to oligonucleotides through covalent linkers. In certain oligomeric compounds, the covalent linker is a single chemical bond (i.e., the conjugate moiety is attached directly to an oligonucleotide through a single bond). In certain oligomeric compounds, a conjugate moiety is attached to an oligonucleotide via a more complex covalent linker comprising one or more covalent linker moieties, which are sub-units making up a covalent linker. In certain aspects, the covalent linker comprises a chain structure, such as a hydrocarbyl chain, or an oligomer of repeating units such as ethylene glycol, nucleosides, or amino acid units.

[0052] In some aspects, the linker links 3’ strand of the first passenger strand with 3’ of the second passenger strand, 3’ strand of the first passenger strand with 5’ of the second passenger strand, 5’ strand of the first passenger strand with 3’ of the second passenger strand, or 5’ strand of the first passenger strand with 5’ of the second passenger strand.301871278 - 15 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0053] In certain aspects, a covalent linker comprises pyrrolidine. In certain aspects, a covalent linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain such aspects, the covalent linker comprises groups selected from alkyl, amino, oxo, amide and ether groups. In certain aspects, the covalent linker comprises groups selected from alkyl and amide groups. In certain aspects, the covalent linker comprises groups selected from alkyl and ether groups. In certain aspects, the covalent linker comprises at least one phosphorus moiety. In certain aspects, the covalent linker comprises at least one phosphate group. In certain aspects, the covalent linker includes at least one neutral linking group.

[0054] In certain aspects, covalent linkers, including the covalent linkers described above, are bifunctional linking moieties, e.g., those known in the art to be useful for attaching conjugate groups to parent compounds, such as the oligonucleotides provided herein. In general, a bifunctional linking moiety comprises at least two functional groups. One of the functional groups is selected to bind to a particular site on a parent compound and the other is selected to bind to a conjugate group. Examples of functional groups used in a bifunctional linking moiety include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In certain aspects, bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.

[0055] Examples of covalent linkers include but are not limited to polyethylene glycol (PEG), tetraethylene Glycol (TEG), a peptide linker, a phosphodiester linker, a phosphorothioate linker, vinyl phosphonate linker, alkylene, polyamide, 1,2,3-triazole from click chemistry, disulfide, or an enzymatically cleavable peptide linkers, pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC) and 6-aminohexanoic acid (AHEX or AHA). Other covalent linkers include but are not limited to substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted C2-C10 alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.

[0056] In certain aspects, covalent linkers comprise 1-10 linker-nucleosides. In certain aspects, covalent linkers comprise 2-5 linker-nucleosides. In certain aspects, covalent linkers comprise exactly 3 linker-nucleosides. In certain aspects, covalent linkers comprise the TCA motif. In certain aspects, such linker-nucleosides are modified nucleosides. In certain aspects such linker-nucleosides comprise a modified sugar moiety. In certain aspects, linker-nucleosides are unmodified. In certain aspects, linker-nucleosides comprise an optionally301871278 - 16 -Atty. Docket Number: UTSDP3820WO- 1001380566protected heterocyclic base selected from a purine, substituted purine, pyrimidine or substituted pyrimidine. In certain aspects, a cleavable moiety is a nucleoside selected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methylcytosine, adenine, 6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine. It is typically desirable for linker-nucleosides to be cleaved from the oligomeric compound after it reaches a target tissue. Accordingly, linker-nucleosides are typically linked to one another and to the remainder of the oligomeric compound through cleavable bonds. In certain aspects, such cleavable bonds are phosphodiester bonds.

[0057] Herein, linker-nucleosides are not considered to be part of the oligonucleotide. Accordingly, in aspects in which an oligomeric compound comprises an oligonucleotide consisting of a specified number or range of linked nucleosides and / or a specified percent complementarity to a reference nucleic acid and the oligomeric compound also comprises a conjugate group comprising a covalent linker comprising linker-nucleosides, those linker-nucleosides are not counted toward the length of the oligonucleotide and are not used in determining the percent complementarity of the oligonucleotide for the reference nucleic acid. For example, an oligomeric compound may comprise (1) a modified oligonucleotide consisting of 8-30 nucleosides and (2) a conjugate group comprising 1-10 linker-nucleosides that are contiguous with the nucleosides of the modified oligonucleotide. The total number of contiguous linked nucleosides in such an oligomeric compound is more than 30. Alternatively, an oligomeric compound may comprise a modified oligonucleotide consisting of 8-30 nucleosides and no conjugate group. The total number of contiguous linked nucleosides in such an oligomeric compound is no more than 30. Unless otherwise indicated covalent linkers comprise no more than 10 linker-nucleosides. In certain aspects, covalent linkers comprise no more than 5 linker-nucleosides. In certain aspects, covalent linkers comprise no more than 3 linker-nucleosides. In certain aspects, covalent linkers comprise no more than 2 linker-nucleosides. In certain aspects, covalent linkers comprise no more than 1 linker-nucleoside.

[0058] In certain aspects, it is desirable for a conjugate group to be cleaved from the oligonucleotide. For example, in certain circumstances oligomeric compounds comprising a particular conjugate moiety are better taken up by a particular cell type, but once the oligomeric compound has been taken up, it is desirable that the conjugate group be cleaved to release the unconjugated or parent oligonucleotide. Thus, certain covalent linkers may comprise one or more cleavable moieties. In certain aspects, a cleavable moiety is a cleavable bond. In certain aspects, a cleavable moiety is a group of atoms comprising at least one cleavable bond. In certain aspects, a cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain aspects, a cleavable moiety is301871278 - 17 -Atty. Docket Number: UTSDP3820WO- 1001380566selectively cleaved inside a cell or subcellular compartment, such as a lysosome. In certain aspects, a cleavable moiety is selectively cleaved by endogenous enzymes, such as nucleases.

[0059] In certain aspects, a cleavable bond is selected from among an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, or a disulfide. In certain aspects, a cleavable bond is one or both of the esters of a phosphodiester. In certain aspects, a cleavable moiety comprises a phosphate or phosphodiester. In certain aspects, the cleavable moiety is a phosphate or phosphodiester linkage between an oligonucleotide and a conjugate moiety or conjugate group.

[0060] In certain aspects, a cleavable moiety comprises or consists of one or more linker-nucleosides. In certain such aspects, the one or more linker-nucleosides are linked to one another and / or to the remainder of the oligomeric compound through cleavable bonds. In certain aspects, such cleavable bonds are unmodified phosphodiester bonds. In certain aspects, a cleavable moiety is 2'-deoxynucleoside that is attached to either the 3' or 5'-terminal nucleoside of an oligonucleotide by a phosphodiester internucleoside linkage and covalently attached to the remainder of the covalent linker or conjugate moiety by a phosphate or phosphorothioate internucleoside linkage. In certain such aspects, the cleavable moiety is 2'-deoxyadenosine.Terminal Groups

[0061] In certain aspects, oligomeric compounds comprise one or more terminal groups. In certain such aspects, oligomeric compounds comprise a stabilized 5'-phosphate. Stabilized 5'-phosphates include, but are not limited to 5'-phosphoanates, including, but not limited to 5'-vinylphosphonates. In certain aspects, terminal groups comprise one or more abasic nucleosides and / or inverted nucleosides. In certain aspects, terminal groups comprise one or more 2'-linked nucleosides. In certain such aspects, the 2'-linked nucleoside is an abasic nucleoside.Conjugated functional moieties

[0062] In other aspects, the disclosed aptamer may be modified with one or more functional moieties. A functional moiety is a molecule that confers one or more additional activities to the aptamer. In certain aspects, the functional moieties enhance cellular uptake by target cells (e.g., neuronal cells). Thus, the invention includes aptamer which are conjugated or unconjugated (e.g., at its 5’ and / or 3’ terminus) to another moiety (e.g. a non-nucleic acid moiety such as a peptide), an organic compound (e.g., a dye), or the like. The conjugation can301871278 - 18 -Atty. Docket Number: UTSDP3820WO- 1001380566be accomplished by methods known in the art, e.g., using the methods of Lambert et al., Drug Deliv. Rev.: 47(1), 99-112 (2001) (describes nucleic acids loaded to polyalkylcyanoacrylate (PACA) nanoparticles); Fattal et al., J. Control Release 53(1 -3): 137-43 (1998) (describes nucleic acids bound to nanoparticles); Schwab et al., Ann. Oncol. 5 Suppl. 4:55-8 (1994) (describes nucleic acids linked to intercalating agents, hydrophobic groups, polycations or PACA nanoparticles); and Godard et al., Eur. J. Biochem.232(2):404-10 (1995) (describes nucleic acids linked to nanoparticles).

[0063] In a certain aspect, the functional moiety is a hydrophobic moiety. In a certain aspect, the hydrophobic moiety is selected from the group consisting of fatty acids, steroids, secosteroids, lipids, gangliosides and nucleoside analogs, endocannabinoids, and vitamins. In a certain aspect, the steroid may be selected from the group consisting of cholesterol and lithocholic acid (LCA). In a certain aspect, the fatty acid selected from the group consisting of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and docosanoic acid (DCA). In some aspects, the vitamin is selected from the group comprising of choline, vitamin A, vitamin E, or derivatives or metabolites thereof. In a certain aspect, the vitamin is selected from the group consisting of retinoic acid and alpha-tocopheryl succinate.

[0064] In a certain aspect, an aptamer of the present disclosure is conjugated to a lipophilic moiety. In one aspect, the lipophilic moiety is a ligand that includes a cationic group. In certain aspects, the lipophilic moiety is selected from the group consisting of cholesterol, vitamin E, vitamin K, vitamin A, folic acid, a cationic dye (e.g., Cy3). In an exemplary aspect, the lipophilic moiety is cholesterol. Other lipophilic moieties include cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-Bis-O(hexadecy1)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, 03-(oleoyl)lithocholic acid, 03-(oleoyl)cholenic acid, dimethoxytrityl, or phenoxazine.

[0065] In certain aspects, the functional moieties may comprise one or more ligands tethered to a aptamer to improve stability, hybridization thermodynamics with a target nucleic acid, targeting to a particular tissue or cell-type, or cell permeability, e.g., by an endocytosis-dependent or -independent mechanism. Ligands and associated modifications can also increase sequence specificity and consequently decrease off-site targeting. A tethered ligand can include one or more modified bases or sugars that can function as intercalators. These can be located in an internal region, such as in a bulge of aptamer / target duplex. The intercalatorcan be an aromatic, e.g., a polycyclic aromatic or heterocyclic aromatic compound. A polycyclic intercalator can have stacking capabilities, and can include systems with 2, 3, or 4 fused rings. The universal bases described herein can be included on a ligand. In one301871278 - 19 -Atty. Docket Number: UTSDP3820WO- 1001380566aspect, the ligand can include a cleaving group that contributes to target gene inhibition by cleavage of the target nucleic acid. The cleaving group can be, for example, a bleomycin (e.g., bleomycin-A5, bleomycin-A2, or bleomycin-B2), pyrene, phenanthroline (e.g., O-phenanthroline), a polyamine, a tripeptide (e.g., lys-tyr-lys tripeptide), ora metal ion chelating group. The metal ion chelating group can include, e.g., an Lu(lll) or Ell(lll) macrocyclic complex, aZn(ll) 2,9-dimethylphenanthroline derivative, a Cu(l I) terpyridine, or acridine, which can promote the selective cleavage of target RNA at the site of the bulge by free metal ions, such as Lu(lll). In some aspects, a peptide ligand can be tethered to a aptamer to promote cleavage of the target RNA, e.g., at the bulge region. For example, l,8-dimethyl-1 ,3,6,8, 10, 13-hexaazacyclotetradecane (cyclam) can be conjugated to a peptide (e.g., by an amino acid derivative) to promote target RNA cleavage. A tethered ligand can be an aminoglycoside ligand, which can cause an aptamer to have improved hybridization properties or improved sequence specificity. Exemplary aminoglycosides include glycosylated polylysine, galactosylated polylysine, neomycin B, tobramycin, kanamycin A, and acridine conjugates of aminoglycosides, such as Neo-N-acridine, Neo-S-acridine, Neo-C-acridine, Tobra-N-acridine, and KanaA-N-acridine. Use of an acridine analog can increase sequence specificity. For example, neomycin B has a high affinity for RNA as compared to DNA, but low sequencespecificity. An acridine analog, neo-5-acridine, has an increased affinity for the HIV Rev-response element (RRE). In some aspects, the guanidine analog (the guanidinoglycoside) of an aminoglycoside ligand is tethered to an aptamer. In a guanidinoglycoside, the amine group on the amino acid is exchanged for a guanidine group. Attachment of a guanidine analog can enhance cell permeability of an aptamer. A tethered ligand can be a poly-arginine peptide, peptoid or peptidomimetic, which can enhance the cellular uptake of an oligonucleotide agent.

[0066] Exemplary ligands are coupled, either directly or indirectly, via an intervening tether, to a ligand-conjugated carrier. In certain aspects, the coupling is through a covalent bond. In certain aspects, the ligand is attached to the carrier via an intervening tether. In certain aspects, a ligand alters the distribution, targeting or lifetime of an aptamer into which it is incorporated. In certain aspects, a ligand provides an enhanced affinity for a selected target, e.g., molecule, cell or cell type, compartment, e.g., a cellular or organ compartment, tissue, organ or region of the body, as, e.g., compared to a species absent such a ligand.

[0067] Exemplary ligands can improve transport, hybridization, and specificity properties and may also improve nuclease resistance of the resultant natural or modified aptamer, or a polymeric molecule comprising any combination of monomers described herein and / or natural or modified ribonucleotides. Ligands in general can include therapeutic modifiers, e.g., for enhancing uptake; diagnostic compounds or reporter groups e.g., for monitoring distribution;301871278 - 20 -Atty. Docket Number: UTSDP3820WO- 1001380566cross-linking agents; nuclease-resistance conferring moieties; and natural or unusual nucleobases. General examples include lipophiles, lipids, steroids (e.g., uvaol, hecigenin, diosgenin), terpenes (e.g., triterpenes, e.g., sarsasapogenin, Friedelin, epifriedelanol derivatized lithocholic acid), vitamins (e.g., folic acid, vitamin A, biotin, pyridoxal), carbohydrates, proteins, protein binding agents, integrin targeting molecules, polycationics, peptides, polyamines, and peptide mimics. Ligands can include a naturally occurring substance, (e.g., human serum albumin (HSA), low-density lipoprotein (LDL), or globulin); carbohydrate (e.g., a dextran, pullulan, chitin, chitosan, inulin, cyclodextrin or hyaluronic acid); amino acid, or a lipid. The ligand may also be a recombinant or synthetic molecule, such as a synthetic polymer, e.g., a synthetic polyamino acid. Examples of polyamino acids include polyamino acid is a polylysine (PLL), poly L-aspartic acid, poly L-glutamic acid, styrene-maleic acid anhydride copolymer, poly(L-lactide-co-glycolied) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydroxypropyl)methacrylamide copolymer (HMPA), polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyurethane, poly(2-ethylacryllic acid), N-isopropylacrylamide polymers, or polyphosphazine. Example of polyamines include: polyethylenimine, polylysine (PLL), spermine, spermidine, polyamine, pseudopeptidepolyamine, peptidomimetic polyamine, dendrimer polyamine, arginine, amidine, protamine, cationic lipid, cationic porphyrin, quaternary salt of a polyamine, or an alpha helical peptide.

[0068] Ligands can also include targeting groups, e.g., a cell or tissue targeting agent, e.g., a lectin, glycoprotein, lipid or protein, e.g., an antibody, that binds to a specified cell type. A targeting group can be a thyrotropin, melanotropin, lectin, glycoprotein, surfactant protein A, mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine (GalNAc) or derivatives thereof, N-acetyl-glucosamine, multivalent mannose, multivalent fucose, glycosylated polyaminoacids, multivalent galactose, transferrin, bisphosphonate, polyglutamate, polyaspartate, a lipid, cholesterol, a steroid, bile acid, folate, vitamin B12, biotin, or an RGD peptide or RGD peptide mimetic. Other examples of ligands include dyes, intercalating agents (e.g. acridines and substituted acridines), cross-linkers (e.g. psoralene, mitomycin C), porphyrins (TPPC4, texaphyrin, Sapphyrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine, phenanthroline, pyrenes), lys-tyr-lys tripeptide, aminoglycosides, guanidium aminoglycodies, artificial endonucleases (e.g. EDTA), lipophilic molecules, e.g, cholesterol (and thio analogs thereof), cholic acid, cholanic acid, lithocholic acid, adamantane acetic acid, 1 -pyrene butyric acid, dihydrotestosterone, glycerol (e.g., esters (e.g., mono, bis, or tris fatty acid esters, e.g., C10, C11 , C12, C13, 014, 01s, 016, 017, 018, 019, or 020 fatty acids) and ethers thereof, e.g., 010, 011, 012, 013, 014, 01s, 016, 017, 018, 019, or 020 alkyl; e.g., 1,3-bis-O(hexadecyl)glycerol, 1,3-bis-O(octaadecyl)glycerol), geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl301871278 - 21 -Atty. Docket Number: UTSDP3820WO- 1001380566group, palmitic acid, stearic acid (e.g., glyceryl distearate), oleic acid, myristic acid, 03-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl, or phenoxazine) and peptide conjugates (e.g., antennapedia peptide, Tat peptide), alkylating agents, phosphate, amino, mercapto, PEG (e.g., PEG-40K), MPEG, [MPEG]2, polyamino, alkyl, substituted alkyl, radiolabeled markers, enzymes, haptens (e.g. biotin), transport / absorption facilitators (e.g., aspirin, naproxen, vitamin E, folic acid), synthetic ribonucleases (e.g., imidazole, bisimidazole, histamine, imidazole clusters, acridine-imidazole conjugates, Eu3+ complexes of tetraazamacrocycles), dinitrophenyl, HRP or AP. In certain aspects, the ligand is GalNAc or a derivative thereof.

[0069] Ligands can be proteins, e.g., glycoproteins, or peptides, e.g., molecules having a specific affinity for a co-ligand, or antibodies e.g., an antibody, that binds to a specified cell type such as a cancer cell, endothelial cell, or bone cell. Ligands may also include hormones and hormone receptors. They can also include non-peptidic species, such as lipids, lectins, carbohydrates, vitamins, cofactors, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine multivalent mannose, or multivalent fucose. The ligand can be, for example, a lipopolysaccharide.

[0070] The ligand can be a substance, e.g., a drug, which can increase the uptake of the aptamer into the cell, for example, by disrupting the cell’s cytoskeleton, e.g., by disrupting the cell’s microtubules, microfilaments, and / or intermediate filaments. The drug can be, for example, taxon, vincristine, vinblastine, cytochalasin, nocodazole, japlakinolide, latrunculin A, phal loidin , swinholide A, indanocine, or myoservin. The ligand can increase the uptake of the aptamer into the cell by activating an inflammatory response, for example. A lipid based ligand can be used to modulate, e.g., control the binding of the conjugate to a target tissue.

[0071] In another aspect, the ligand is a moiety, e.g., a vitamin, which is taken up by a target cell, e.g., a proliferating cell. These can be useful for treating disorders characterized by unwanted cell proliferation, e.g., of the malignant or non-malignant type, e.g., cancer cells. Exemplary vitamins include vitamin A, E, and K. Other exemplary vitamins include are B vitamin, e.g., folic acid, B12, riboflavin, biotin, pyridoxal or other vitamins or nutrients taken up by cancer cells. Also included are HSA and low-density lipoprotein (LDL).

[0072] In another aspect, the ligand is a cell-permeation agent, such as a helical cellpermeation agent. In certain aspects, the agent is amphipathic. An exemplary agent is a peptide such as tat or antennopedia. If the agent is a peptide, it can be modified, including a peptidylmimetic, invertomers, non-peptide or pseudo-peptide linkages, and use of D-amino acids. The helical agent can be an alpha-helical agent, which may have a lipophilic and a lipophobic phase.301871278 - 22 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0073] The ligand can be a peptide or peptidomimetic. A peptidomimetic (also referred to herein as an oligopeptidomimetic) is a molecule capable of folding into a defined three-dimensional structure similar to a natural peptide. The attachment of peptide and peptidomimetics to oligonucleotide agents can affect pharmacokinetic distribution, such as by enhancing cellular recognition and absorption. The peptide or peptidomimetic moiety can be about 5-50 amino acids long, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids long. A peptide or peptidomimetic can be, for example, a cell permeation peptide, cationic peptide, amphipathic peptide, or hydrophobic peptide (e.g., consisting primarily of Tyr, Trp or Phe). The peptide moiety can be a dendrimer peptide, constrained peptide or crosslinked peptide. The peptide moiety can be an L-peptide or D-peptide. In another alternative, the peptide moiety can include a hydrophobic membrane translocation sequence (MTS). A peptide or peptidomimetic can be encoded by a random sequence of DNA, such as a peptide identified from a phage-display library, or one-bead-one-compound (OBOC) combinatorial library (Lam et al., Nature 354:82-84, 1991). In exemplary aspects, the peptide or peptidomimetic tethered to an aptamer via an incorporated monomer unit is a cell targeting peptide such as an arginine-glycine-aspartic acid (RGD)-peptide, or RGD mimic. A peptide moiety can range in length from about 5 amino acids to about 40 amino acids. The peptide moieties can have a structural modification, such as to increase stability or direct conformational properties. Any of the structural modifications described below can be utilized.IV. Compositions

[0074] In some aspects, the current disclosure also encompasses compositions comprising one or more of the disclosed aptamers, or variants thereof. Compositions, in particular pharmaceutical compositions of the present invention can include, without limitation, saline, buffered solutions, lipidoids, liposomes, lipid nanoparticles, polymers, lipoplexes, core-shell nanoparticles, cells transfected with viral vectors (e.g., for transplantation into a subject), nanoparticle mimics and combinations thereof. In some aspects, the compositions disclosed herein are chemical vectors, such as a lipid complex or naked DNA. “Naked DNA” or “naked nucleic acid” refers to a nucleic acid molecule that is not contained in encapsulating means that facilitates delivery of a nucleic acid into the cytoplasm of a target host cell. Naked DNA may be circular or linear (linearized DNA sequence). Optionally, a naked nucleic acid can be associated with standard means used in the art for facilitating its delivery of the nucleic acid to the target host cell, for example to facilitate the transport of the nucleic acid through the cell membrane. In some aspects, the composition is a pharmaceutical composition comprising one or more excipients as disclosed herein below.301871278 - 23 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0075] A pharmaceutical composition in accordance with the present disclosure may be prepared, packaged, and / or sold in bulk, as a single unit dose, and / or as a plurality of single unit doses. As used herein, a “unit dose” refers to a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient (for example, one or more anti-LEAP2 aptamers alone or in combination with one or more anti-LEAP2 shRNAs, and / or Ghrelin receptor agonist). The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and / or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

[0076] Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and / or any additional ingredients in a pharmaceutical composition in accordance with the present disclosure may vary, depending upon the identity, size, and / or condition of the subject being treated and further depending upon the route by which the composition is to be administered. For example, the composition may comprise between 0.1% and 99% (w / w) of the active ingredient. By way of example, the composition may comprise between 0.1% and 100%, e.g., between 0.5 and 50%, between 1-30%, between 5-80%, at least 80% (w / w) active ingredient.

[0077] In some aspects, a pharmaceutically acceptable excipient may be at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some aspects, an excipient is approved for use for humans and for veterinary use. In some aspects, an excipient may be approved by United States Food and Drug Administration. In some aspects, an excipient may be of pharmaceutical grade. In some aspects, an excipient may meet the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeia.

[0078] In certain aspects, compositions disclosed herein may comprise one or more pharmaceutically acceptable excipient(s), diluents, and / or carrier(s). As used herein, a pharmaceutically acceptable diluent, excipient, or carrier, refers to a material suitable for administration to a subject without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained. Pharmaceutically acceptable diluents, carriers, and excipients can include, but are not limited to, physiological saline, Ringer’s solution, phosphate solution or buffer, buffered saline, and other carriers known in the art.

[0079] In some aspects, pharmaceutical compositions herein may include stabilizers, antioxidants, colorants, other medicinal or pharmaceutical agents, carriers, adjuvants, preserving agents, stabilizing agents, wetting agents, emulsifying agents, solution promoters, salts, solubilizers, antifoaming agents, antioxidants, dispersing agents, surfactants, or any301871278 - 24 -Atty. Docket Number: UTSDP3820WO- 1001380566combination thereof. Herein, the term “excipient” refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols. Techniques for formulation and administration of drugs may be found in “Remington’s Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., latest edition, which is incorporated herein by reference.

[0080] In certain aspects, pharmaceutical compositions described herein may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries to facilitate processing of genetically modified endothelial progenitor cells into preparations which can be used pharmaceutically. In some aspects, any of the well-known techniques, carriers, and excipients may be used as suitable and / or as understood in the art.

[0081] In certain aspects, pharmaceutical compositions described herein may be an aqueous suspension comprising one or more polymers as suspending agents. In some aspects, polymers that may comprise pharmaceutical compositions described herein include: water-soluble polymers such as cellulosic polymers, e.g., hydroxypropyl methylcellulose; waterinsoluble polymers such as cross-linked carboxyl-containing polymers; mucoadhesive polymers, selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran; or a combination thereof. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% total amount of polymers as suspending agent(s) by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of polymers as suspending agent(s) by total weight of the composition.

[0082] In certain aspects, pharmaceutical compositions disclosed herein may comprise a viscous formulation. In some aspects, viscosity of composition herein may be increased by the addition of one or more gelling or thickening agents. In some aspects, compositions disclosed herein may comprise one or more gelling or thickening agents in an amount to provide a sufficiently viscous formulation to remain on treated tissue. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% total amount of gelling or thickening301871278 - 25 -Atty. Docket Number: UTSDP3820WO- 1001380566agent(s) by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of gelling or thickening agent(s) by total weight of the composition. In some aspects, suitable thickening agents for use herein can be hydroxypropyl methylcellulose, hydroxyethyl cellulose, polyvinylpyrrolidone, carboxymethyl cellulose, polyvinyl alcohol, sodium chondroitin sulfate, sodium hyaluronate. In other aspects, viscosity enhancing agents can be acacia (gum arabic), agar, aluminum magnesium silicate, sodium alginate, sodium stearate, bladderwrack, bentonite, carbomer, carrageenan, Carbopol, xanthan, cellulose, microcrystalline cellulose (MCC), ceratonia, chitin, carboxymethylated chitosan, chondrus, dextrose, furcellaran, gelatin, Ghatti gum, guar gum, hectorite, lactose, sucrose, maltodextrin, mannitol, sorbitol, honey, maize starch, wheat starch, rice starch, potato starch, gelatin, sterculia gum, xanthum gum, gum tragacanth, ethyl cellulose, ethyl hydroxyethyl cellulose, ethylmethyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxyethylmethyl cellulose, hydroxypropyl cellulose, poly(hydroxyethyl methacrylate), oxypolygelatin, pectin, polygeline, povidone, propylene carbonate, methyl vinyl ether / maleic anhydride copolymer (PVM / MA), poly(methoxyethyl methacrylate), poly(methoxyethoxyethyl methacrylate), hydroxypropyl cellulose, hydroxypropylmethyl-cellulose (HPMC), sodium carboxymethyl-cellulose (CMC), silicon dioxide, polyvinylpyrrolidone (PVP: povidone), Splenda (dextrose, maltodextrin and sucralose), or any combination thereof.

[0083] In certain aspects, pharmaceutical compositions disclosed herein may comprise additional agents or additives selected from a group including surface-active agents, detergents, solvents, acidifying agents, alkalizing agents, buffering agents, tonicity modifying agents, ionic additives effective to increase the ionic strength of the solution, antimicrobial agents, antibiotic agents, antifungal agents, antioxidants, preservatives, electrolytes, antifoaming agents, oils, stabilizers, enhancing agents, and the like. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% total amount of one or more agents by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more agents by total weight of the composition. In some aspects, one or more of these agents may be added to improve the performance, efficacy, safety, shelflife and / or other property of the muscarinic antagonist composition of the present disclosure. In some aspects, additives may be biocompatible, without being harsh, abrasive, and / or allergenic.301871278 - 26 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0084] In certain aspects, pharmaceutical compositions disclosed herein may comprise one or more acidifying agents. As used herein, “acidifying agents” refers to compounds used to provide an acidic medium. Such compounds include, by way of example and without limitation, acetic acid, amino acid, citric acid, fumaric acid and other alpha hydroxy acids, such as hydrochloric acid, ascorbic acid, and nitric acid and others known to those of ordinary skill in the art. In some aspects, any pharmaceutically acceptable organic or inorganic acid may be used. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more acidifying agents by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more acidifying agents by total weight of the composition.

[0085] In certain aspects, pharmaceutical compositions disclosed herein may comprise one or more alkalizing agents. As used herein, “alkalizing agents” are compounds used to provide alkaline medium. Such compounds include, by way of example and without limitation, ammonia solution, ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium borate, sodium carbonate, sodium bicarbonate, sodium hydroxide, triethanolamine, and trolamine and others known to those of ordinary skill in the art. In some aspects, any pharmaceutically acceptable organic or inorganic base can be used. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more alkalizing agents by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more alkalizing agents by total weight of the composition.

[0086] In certain aspects, pharmaceutical compositions disclosed herein may comprise one or more antioxidants. As used herein, “antioxidants” are agents that inhibit oxidation and thus can be used to prevent the deterioration of preparations by the oxidative process. Such compounds include, by way of example and without limitation, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorous acid, monothioglycerol, propyl gallate, sodium ascorbate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfite and other materials known to one of ordinary skill in the art. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least301871278 - 27 -Atty. Docket Number: UTSDP3820WO- 1001380566about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more antioxidants by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more antioxidants by total weight of the composition.

[0087] In certain aspects, pharmaceutical compositions disclosed herein may comprise a buffer system. As used herein, a “buffer system” is a composition comprised of one or more buffering agents wherein “buffering agents” are compounds used to resist change in pH upon dilution or addition of acid or alkali. Buffering agents include, by way of example and without limitation, potassium metaphosphate, potassium phosphate, monobasic sodium acetate and sodium citrate anhydrous and dihydrate and other materials known to one of ordinary skill in the art. In some aspects, any pharmaceutically acceptable organic or inorganic buffer can be used. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more buffering agents by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more buffering agents by total weight of the composition.

[0088] In some aspects, the amount of one or more buffering agents may depend on the desired pH level of a composition. In some aspects, pharmaceutical compositions disclosed herein may have a pH of about 6 to about 9. In some aspects, pharmaceutical compositions disclosed herein may have a pH greater than about 8, greater than about 7.5, greater than about 7, greater than about 6.5, or greater than about 6.

[0089] In certain aspects, pharmaceutical compositions disclosed herein may comprise one or more preservatives. As used herein, “preservatives” refers to agents or combination of agents that inhibits, reduces or eliminates bacterial growth in a pharmaceutical dosage form. Non-limiting examples of preservatives include Nipagin, Nipasol, isopropyl alcohol and a combination thereof. In some aspects, any pharmaceutically acceptable preservative can be used. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more preservatives by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more preservatives by total weight of the composition.301871278 - 28 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0090] In certain aspects, pharmaceutical compositions disclosed herein may comprise one or more surface-acting reagents or detergents. In some aspects, surface-acting reagents or detergents may be synthetic, natural, or semi-synthetic. In some aspects, compositions disclosed herein may comprise anionic detergents, cationic detergents, zwitterionic detergents, ampholytic detergents, amphoteric detergents, nonionic detergents having a steroid skeleton, or a combination thereof. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more surface-acting reagents or detergents by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more surface-acting reagents or detergents by total weight of the composition.

[0091] In certain aspects, pharmaceutical compositions disclosed herein may comprise one or more stabilizers. As used herein, a “stabilizer” refers to a compound used to stabilize an active agent against physical, chemical, or biochemical process that would otherwise reduce the therapeutic activity of the agent. Suitable stabilizers include, by way of example and without limitation, succinic anhydride, albumin, sialic acid, creatinine, glycine and other amino acids, niacinamide, sodium acetyltryptophonate, zinc oxide, sucrose, glucose, lactose, sorbitol, mannitol, glycerol, polyethylene glycols, sodium caprylate, and sodium saccharin and others known to those of ordinary skill in the art. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more stabilizers by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more stabilizers by total weight of the composition.

[0092] In some aspects, pharmaceutical compositions disclosed herein may comprise one or more tonicity agents. As used herein, a “tonicity agents” refers to a compound that can be used to adjust the tonicity of the liquid formulation. Suitable tonicity agents include, but are not limited to, glycerin, lactose, mannitol, dextrose, sodium chloride, sodium sulfate, sorbitol, trehalose and others known to those or ordinary skill in the art. Osmolarity in a composition may be expressed in milliosmoles per liter (mOsm / L). Osmolarity may be measured using methods commonly known in the art. In some aspects, a vapor pressure depression method is used to calculate the osmolarity of the compositions disclosed herein. In some aspects, the amount of one or more tonicity agents comprising a pharmaceutical composition disclosed301871278 - 29 -Atty. Docket Number: UTSDP3820WO- 1001380566herein may result in a composition osmolarity of about 150 mOsm / L to about 500 mOsm / L, about 250 mOsm / L to about 500 mOsm / L, about 250 mOsm / L to about 350 mOsm / L, about 280 mOsm / L to about 370 mOsm / L or about 250 mOsm / L to about 320 mOsm / L. In some aspects, a composition herein may have an osmolality ranging from about 100 mOsm / kg to about 1000 mOsm / kg, from about 200 mOsm / kg to about 800 mOsm / kg, from about 250 mOsm / kg to about 500 mOsm / kg, or from about 250 mOsm / kg to about 320 mOsm / kg, or from about 250 mOsm / kg to about 350 mOsm / kg or from about 280 mOsm / kg to about 320 mOsm / kg. In some aspects, a pharmaceutical composition described herein may have an osmolarity of about 100 mOsm / L to about 1000 mOsm / L, about 200 mOsm / L to about 800 mOsm / L, about 250 mOsm / L to about 500 mOsm / L, about 250 mOsm / L to about 350 mOsm / L, about 250 mOsm / L to about 320 mOsm / L, or about 280 mOsm / L to about 320 mOsm / L. In some aspects, pharmaceutical compositions disclosed herein may comprise at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% total amount of one or more tonicity modifiers by total weight of the composition. In some aspects, pharmaceutical compositions disclosed herein may comprise about 5% to about 99%, about 10%, about 95%, or about 15% to about 90% total amount of one or more tonicity modifiers by total weight of the composition.V. Delivery particles

[0093] In some aspects, the composition as disclosed herein may comprise a delivery particle. The delivery particle compositions may encapsulate or be associated with one or more active agents (one or more anti-LEAP2 aptamers, alone or in combination with one or more anti-LEAP2 shRNAs, and / or GHSR agonists), which may be, but are not limited to: polynucleotides, engineered protein compositions, and / or small molecules. In some aspects, polynucleotides, engineered protein compositions, and / or small molecules formulated in the delivery particles can have improved pharmacokinetic and / or pharmacodynamic properties. In some aspects, polynucleotides, engineered protein compositions, and / or small molecules delivery particles are better tolerated by a patient. In some aspects, polynucleotides, engineered protein compositions, and / or small molecules are formulated in the delivery particles, and in some aspects, are more effectively delivered to cells to affect their function, than naked engineered proteins, polynucleotides, and / or small molecules. In some aspects, a delivery particle composition confers water solubility to hydrophobic agents, to combinations of hydrophobic agents, and / or to combinations of hydrophobic and hydrophilic agents. In some aspects, a delivery particle composition comprises a liposomal and / or nano-emulsion composition of a therapeutic agent. In some aspects, the delivery particle is a nanoparticle or a microparticle.301871278 - 30 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0094] In several aspects, as disclosed elsewhere herein, a nanoparticle composition (e.g., a mixed micelle composition, a liposomal composition, solid lipid particles, oil-in-water emulsions, water-in-oil-in-water emulsions, water-in-oil emulsions, oil-in-water-in-oil emulsions, etc.) is provided to aid in the delivery of therapeutic agents. In certain aspects, pharmaceutical compositions described herein may be formulated in the form of a nanoparticle. The nanoparticle may have a monolayer enclosing the nanoparticle core, wherein the polynucleotide construct is disposed within the nanoparticle core. In an aspect, the nanoparticle core includes a solid lipid (i.e., lipid that remains solid at room temperature and body temperature) or a liquid lipid (i.e., oil, which remains liquid at room temperature and body temperature, for example, vegetable oil or a lipid extracted from human adipose tissue). In particular, aspects of the present disclosure include nanoparticles and compositions for the controlled and / or sustained release (e.g., release at a predetermined rate to maintain a certain concentration for a certain period of time) of an agent, such as a polynucleotide construct from the nanoparticle. In some aspects, the nanoparticles comprise one or more therapeutic agents. In some aspects, a composition comprising the nanoparticles disclosed herein comprises a therapeutically effective amount of one or more therapeutic agents.

[0095] In several aspects, when formulated, the dry weight % of one or more therapeutic agents present in the nanoparticle compositions is equal to or at least about: 0.1%, 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 12.5%, 15%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5%, 50%, 52.5%, 55%, 57.5%, 60%, or any range derivable therein. In several aspects, the therapeutic agents are provided in an aqueous composition. In several aspects, the wet weight % of one or more therapeutic agents present in the composition (with water included) is equal to or at least about: 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 12.5%, 15%, 20%, 22.5%, 25%, 27.5%, 30%, or any range derivable therein. In several aspects, the one or more therapeutic agents may be provided in the wet composition at a concentration of greater than or equal to about: 0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 5 mg / mL, 20 mg / mL, 30 mg / mL, 50 mg / mL, 100 mg / mL, or any range derivable therein.

[0096] In several aspects, the therapeutic agents, collectively or individually, are present in the aqueous nanoparticle composition at a concentration of less than or equal to about: 150 mg / mL, 100 mg / mL, 75 mg / mL, 50 mg / mL, 25 mg / mL, 20 mg / mL, 10 mg / mL, 5 mg / mL, 2.5 mg / mL, 2 mg / mL, 1.5 mg / mL, 1 mg / mL, 0.5 mg / mL, 0.1 mg / mL, 0.05 mg / mL, 0.01 mg / mL, or ranges including and / or spanning the aforementioned values. In several aspects, the one or more therapeutic agents, collectively or individually, are present in the aqueous composition at a concentration of greater than or equal to about: 150 mg / mL, 100 mg / mL, 75 mg / mL, 50 mg / mL, 25 mg / mL, 20 mg / mL, 10 mg / mL, 5 mg / mL, 2.5 mg / mL, 2 mg / mL, 1.5 mg / mL, 1301871278 - 31 -Atty. Docket Number: UTSDP3820WO- 1001380566mg / mL, 0.5 mg / mL, 0.1 mg / mL, 0.05 mg / mL, 0.01 mg / mL, or ranges including and / or spanning the aforementioned values. In several aspects, the one or more therapeutic agents, collectively or individually, are present in the composition at a dry wt. % of equal to or at least about: 0.1%, 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 12.5%, 15%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5%, 50%, 52.5%, 55%, 57.5%, 60%, or ranges including and / or spanning the aforementioned values. In several aspects, the one or more therapeutic agents, collectively or individually, are present in the composition at a wet wt. % of equal to or at least about: 0.1%, 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 12.5%, 15%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5%, 50%, 52.5%, 55%, 57.5%, 60%, or ranges including and / or spanning the aforementioned values. In several aspects, as disclosed elsewhere herein, the composition is aqueous, while in others it has been dried into a powder (that is free of or substantially free of water). In several aspects, where the composition has been dried, it comprises a water content of less than or equal to 20%, 15%, 10%, 7.5%, 5%, 2.5%, 1%, or ranges including and / or spanning the aforementioned values.

[0097] In several aspects, as disclosed elsewhere herein, the composition is aqueous (e.g., contains water) while in other aspects, the composition is dry (lacks water or substantially lacks water). In several aspects, the composition has been dried (e.g., has been subjected to a process to remove most or substantially all water). In several aspects, the composition comprises nanoparticles in water (e.g., as a solution, suspension, or emulsion). In other aspects, the composition is provided as a powder (e.g., that may be constituted or reconstituted in water). In several aspects, as disclosed elsewhere herein, the water content (in wt. %) of the composition is less than or equal to about: 30%, 20%, 10%, 5%, 2.5%, 1%, 0.5%, 0.1%, 0%, or ranges including and / or spanning the aforementioned values. In several aspects, as disclosed elsewhere herein, the water content (in wt. %) of the composition is greater than or equal to about: 50%, 60%, 70%, 80%, 85%, 90%, 92.5%, 95%, 97.5%, or ranges including and / or spanning the aforementioned values. In several aspects, the water is nanopure, deionized, USP grade, WFI, and / or combinations of the foregoing. In some aspects, the composition is a dried composition comprising a nanoparticle having weight ratios of a first therapeutic agent: a nanoscale coordination polymer NCP: optionally a lipid source, and optionally a surfactant of 1 to 50:1 to 50:1 to 50:0 to 17.5.

[0098] In some aspects, a nanoparticle composition provides an oil-in-water emulsion (e.g., a nanoemulsion), water-in-oil emulsion, a water-in-oil-in-water emulsion, an oil-in-water-in-oil emulsion, a liposome (and variants including multi-lamellar, double liposome preparations, etc.), micelle, and / or solid lipid particles. Any one of these structures may be provided as a nanoparticle or microparticle.301871278 - 32 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0099] In some aspects, the nanoparticle composition comprises a lipid source. In some aspects, the lipid source comprises a charged lipid, which can impart a charge to the nanoparticle. In some aspects, the lipid source comprises a neutral lipid. In some aspects, the lipid source comprises one or more phospholipids. In some aspects, the one or more phospholipids comprises one or more of phosphatidic acid, phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylinositol phosphate, phosphatidylinositol bisphosphate, phosphatidylinositol trisphosphate, lipoid H 100-3, phospholipon 90H, phospholipon 80H, lipoid 100-3, lipoid P75-3, 1,2-dioleoyl-sn-glycero-3-phosphate (DOPA), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-diastearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)2000] (DSPE-PEG2000) or any combination of the foregoing. In some aspects, the lipid source is a phosphatidylcholine. In some aspects, the one or more lipid source lipid(s) (collectively or individually) are present in the composition at a dry wt. % of equal to or less than about: 0%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, or ranges including and / or spanning the aforementioned values. In some aspects, the one or more lipid source lipid(s) (collectively or individually) are present in the composition at a wet wt. % of equal to or less than about: 0%, 0.1%, 0.5%, 1.0%, 2.5%, 4%, 5%, 6%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or ranges including and / or spanning the aforementioned values. In some aspects, the one or more lipid source lipid(s) (collectively or individually) are present in the composition at a wet w / v of equal to or less than about: 0 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 2.5 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7.5 mg / mL, 10 mg / mL, 12.5 mg / mL, 15 mg / mL, 17.5 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, or ranges including and / or spanning the aforementioned values. In some aspects, the composition is aqueous, while in others it has been dried into a powder. For instance, in several aspects, the composition is aqueous (wet), while in others it has been dried into a powder (dry). In some aspects, the one or more lipid(s) of the lipid source are synthetic, derived from sunflower, soy, egg, or mixtures thereof. In some aspects, the one or more lipids of the lipid source can be hydrogenated or non-hydrogenated. In some aspects, the lipid source exceeds requirements of the United States Pharmacopeia (is USP grade) and / or is National Formulary (NF) grade.

[0100] In some aspects, the one or more lipids of the lipid source has a purity of greater than or equal to about: 92.5%, 95%, 96%, 96.3%, 98%, 99%, 100%, or ranges including and / or spanning the aforementioned values. In several aspects, the one or more lipids of the lipid source has a total % impurity content by weight of less than or equal to about: 8.5%, 5%, 4%, 3.7%, 2%, 1%, 0%, or ranges including and / or spanning the aforementioned values.301871278 - 33 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0101] In some aspects, a nanoparticle composition comprises a surfactant. In certain aspects, a nanoparticle composition does not comprise a surfactant. In some aspects, the surfactant is a pharmaceutically acceptable surfactant. In some aspects, the surfactant is a food surfactant. In some aspects, the surfactant comprises one or more of a polyoxyethylene sorbitan esters (e.g., polysorbates / tweens, including polysorbate 80, polysorbate 20, etc.), cremophor (e.g., a non-ionic solubilizer and emulsifier that is made by reacting ethylene oxide with castor oil), propylene oxide-modified polymethylsiloxane, dodecyl betaine, lauramidopropyl betaine, cocoamido-2-hydroxypropyl sulfobetaine, sodium stearate (or other stearate salts), polyoxyethylene alcohol, lecithins, mono- and diglycerides of fatty acids (MDG), acetic acid esters of MDG, lactic acid esters of MDG, citric acid esters of MDG, mono-and diacetyl tartaric acid esters of MDG, sucrose esters of fatty acids, polyglycerol esters of fatty acids (e.g., polyglycerol esters), polyglycerol polyricinoleate, propane-1, 2-diol esters of fatty acids, propylene glycol esters, sodium stearoyl-2-lactylate, calcium stearoyl-2-lactylate, sorbitan fatty acid esters, quillaja extract surfactant, yucca extract surfactant, saponins, silicone emulsifiers, sorbitan trioleate, soya lecithin, dioctyl sodium sulfosuccinate, dioctyl sodium sulfonate, polyoxyethylene, hydrogenated castor oil, sucrose fatty acid ester, or combinations of any of the foregoing. Natural or synthetic surfactants can be used, including polyethylene glycol and dextrans, such as cyclodextran. In some aspects, the one or more surfactants are present in a nanoparticle composition (collectively or individually) at a dry wt. % of equal to or less than about: 0%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, or ranges including and / or spanning the aforementioned values. In some aspects, surfactants can include cationic, anionic, non-ionic, and zwitterionic surfactants. In some aspects, one or more surfactants (collectively or individually) are present in the composition at a wet wt. % of equal to or less than about: 0%, 0.1%, 0.5%, 1.0%, 2.5%, 4%, 5%, 6%, 7.5%, 10%, 12.5%, 15%, 17.5%, or ranges including and / or spanning the aforementioned values. In some aspects, one or more surfactants (collectively or individually) are present in the composition at a wet w / v of equal to or less than about: 0 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 2.5 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7.5 mg / mL, 10 mg / mL, 12.5 mg / mL, 15 mg / mL, 17.5 mg / mL, or ranges including and / or spanning the aforementioned values. In some aspects, a surfactant exceeds requirements of the United States Pharmacopeia (is USP grade) and / or is National Formulary (NF) grade.

[0102] In several aspects, one or more co-emulsifiers are used. In certain aspects, a nanoparticle composition does not comprise a co-emulsifier. In some aspects, a co-emulsifier is a pharmaceutically acceptable co-emulsifier. In some aspects, a co-emulsifier is selected from the group consisting of oleic acid, miglyol 812N (all versions), cetearyl olivate, isoprpyle myristate, celluloses, polysaccharides (e.g., methylcellulose, propylmethylcellulose,301871278 - 34 -Atty. Docket Number: UTSDP3820WO- 1001380566hydroxypropyl methylcellulose, xanthan gum, etc.), capric acid, caprylic acid, triglycerides (e.g., triglycerides of oleic acid, capric acid, caprylic acid (Captex 8000, Captex GTO, Captex 1000)), glycerol monooleate, glyceryl stearate, glycerol monostearate (Geleol™ Mono and Diglyceride NF), omega-3 fatty acids (a-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), Tonalin, Pronova Pure® 46:38, free fatty acid Tonalin FFA 80), conjugated linoleic acid (CLA), alpha glycerylphosphorylcholine (alpha GPC), palmitoylethanolamide (PEA), cetyl alcohol, or emulsifying wax and / or combinations of any of the foregoing. In some aspects, one or more co-emulsifiers are present in the nanoparticle composition (collectively or individually) at a dry wt. % of equal to or less than about: 0%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, or ranges including and / or spanning the aforementioned values. In some aspects, one or more co-emulsifiers (collectively or individually) are present in the composition at a wet wt. % of equal to or less than about: 0%, 0.1%, 0.5%, 1.0%, 2.5%, 4%, 5%, 6%, 7.5%, 10%, 12.5%, 15%, 17.5%, or ranges including and / or spanning the aforementioned values. In some aspects, one or more co-emulsifiers (collectively or individually) are present in the composition at a wet w / v of equal to or less than about: 0 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 2.5 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7.5 mg / mL, 10 mg / mL, 12.5 mg / mL, 15 mg / mL, 17.5 mg / mL, or ranges including and / or spanning the aforementioned values. In some aspects, a co-emulsifiers exceeds requirements of the United States Pharmacopeia (is USP grade) and / or is National Formulary (NF) grade. In some aspects, a co-emulsifier component comprises a medium chain triglyceride (MCT). In some aspects, a medium chain triglyceride comprises a fatty acid selected from one or more of caprioc acid, octanoic acid, capric acid, caprylic acid, and / or lauric acid (e.g., is formed from). In some aspects, a medium chain triglyceride comprises a fatty acid 6-12 carbons in length (e.g., 6, 7, 8, 9, 10, 11, or 12). In some aspects, a co-emulsifier component comprises a long chain triglyceride (LCT). In some aspects, a long chain triglyceride comprises a fatty acid greater than 12 carbons in length (e.g., greater than or equal to 13, 14, 15, 16, 17, 18, 19, or 20 carbons in length, or ranges including and / or spanning the aforementioned values). In some aspects, a co-emulsifier component is a single lipid. In some aspects, a co-emulsifier component is highly pure. In some aspects, a co-emulsifier component has a purity by weight % of equal to or greater than about: 90%, 95%, 97%, 98%, 99%, 100%, or ranges including and / or spanning the aforementioned values. In some aspects, a co-emulsifier component is present in the nanoparticle composition at dry weight % of equal to or greater than about: 10%, 20%, 30%, 35%, 40%, 45%, 50%, or ranges including and / or spanning the aforementioned values.

[0103] In some aspects, a nanoparticle composition comprises one or more sterols. In certain aspects, a nanoparticle composition does not comprise a sterol. In some aspects, one or more301871278 - 35 -Atty. Docket Number: UTSDP3820WO- 1001380566sterols comprises one or more cholesterols, ergosterols, hopanoids, hydroxysteroids, phytosterols (e.g., vegapure), ecdysteroids, and / or steroids. In some aspects, a sterol comprises a cholesterol. In some aspects, a sterol component is a single sterol. In some aspects, a sterol component is cholesterol. In some aspects, a cholesterol (or other sterol) is highly pure. In some aspects, one or more sterol(s) (e.g., cholesterol, and / or other sterols), collectively or individually, are present in the aqueous composition at a concentration of less than or equal to about: 50 mg / mL, 40 mg / mL, 20 mg / mL, 10 mg / mL, 5 mg / mL, or ranges including and / or spanning the aforementioned values. In some aspects, one or more sterol(s) are present in the composition at a dry wt. % of equal to or less than about: 0.25%, 0.5%, 1%, 5%, 7.5%, 10%, 15%, 20%, 25%, or ranges including and / or spanning the aforementioned values. In some aspects, one or more sterol(s) (collectively or individually) are present in the composition at a wet wt. % of equal to or less than about: 0.1%, 0.25%, 0.5%, 1%, 2%, 3%, 4%, 5%, 7.5%, 10%, or ranges including and / or spanning the aforementioned values. In some aspects, cholesterol used in the composition comprises cholesterol from one or more of sheep’s wool, synthetic cholesterol, or semisynthetic cholesterol from plant origin. In some aspects, a sterol has a purity of greater than or equal to about: 92.5%, 95%, 96%, 98%, 99%, 99.9%, 100.0%, or ranges including and / or spanning the aforementioned values. In some aspects, a sterol has a total % impurity content by weight of less than or equal to about: 8.5%, 5%, 4%, 3.7%, 2%, 1%, 0%, or ranges including and / or spanning the aforementioned values. In some aspects, a sterol is not cholesterol.

[0104] In some aspects, a nanoparticle composition comprises a preservative. In certain aspects, a nanoparticle composition does not comprise a preservative. In several aspects, a preservative includes one or more benzoates (such as sodium benzoate or potassium benzoate), nitrites (such as sodium nitrite), sulfites (such as sulfur dioxide, sodium or potassium sulphite, bisulphite or metabisulphite), sorbates (such as sodium sorbate, potassium sorbate), ethylenediaminetetraacetic acid (EDTA) (and / or the disodium salt thereof), polyphosphates, organic acids (e.g., citric, succinic, malic, tartaric, benzoic, lactic and propionic acids), and / or antioxidants (e.g., vitamins such as vitamin E and / or vitamin C, butylated hydroxytoluene). In several aspects, sorbates and benzoates may be used in acidic pH formulations. In several aspects, one or more preservatives (collectively or individually) are present in the composition at a dry wt. % of equal to or at less than about: 0.01 %, 0.1 %, 0.25%, 0.5%, 1%, 5%, 7.5%, 10%, 15%, 20%, 25%, or ranges including and / or spanning the aforementioned values. In several aspects, one or more preservatives (collectively or individually) are present in the composition at a wet wt. % of equal to or less than about: 0.001%, 0.01%, 0.025%, 0.05%, 0.1%, 0.5%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 5%, or ranges including and / or spanning the aforementioned values. In several aspects, one or more301871278 - 36 -Atty. Docket Number: UTSDP3820WO- 1001380566preservatives (collectively or individually) are present in the composition at a wet w / v of equal to or less than about: 0 mg / mL, 0.001 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 2.5 mg / mL, 4 mg / mL, 5 mg / mL, or ranges including and / or spanning the aforementioned values. In several aspects, as disclosed elsewhere herein, the composition is aqueous, while in others it has been dried into a powder. For instance, as disclosed elsewhere herein, in several aspects, the composition is aqueous (wet), while in others it has been dried into a powder (dry). In several aspects, preservatives inhibit or prevent growth of mold, bacteria, and / or fungus.

[0105] In some aspects, a nanoparticle composition comprises a metal. In some aspects, a metal may be zinc. In certain aspects, a nanoparticle composition does not comprise a metal. In several aspects, one or more metals (collectively or individually) are present in the composition at a dry wt. % of equal to or at less than about: 0.01%, 0.1%, 0.25%, 0.5%, 1%, 5%, 7.5%, 10%, 15%, 20%, 25%, or ranges including and / or spanning the aforementioned values. In several aspects, one or more metals (collectively or individually) are present in the composition at a wet wt. % of equal to or less than about: 0.001%, 0.01%, 0.025%, 0.05%, 0.1%, 0.5%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 5%, or ranges including and / or spanning the aforementioned values. In several aspects, one or more metals (collectively or individually) are present in the composition at a wet w / v of equal to or less than about: 0 mg / mL, 0.001 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 2.5 mg / mL, 4 mg / mL, 5 mg / mL, or ranges including and / or spanning the aforementioned values. In some aspects, metals may be combined with metalloligands to generate a nanoscale coordination polymer. In some aspects, nanoscale coordination polymers may comprise metal-connecting points and organic bridging ligands. In some aspects, a nanoscale coordination polymers (NCP) may self-assemble into nanoparticles.

[0106] In several aspects, as disclosed elsewhere herein, a nanoparticle composition provides particles in the nano-measurement range. In several aspects, a nanoparticle is spherical or substantially spherical. In several aspects, a solid lipid nanoparticle possesses a solid lipid core matrix that can solubilize lipophilic molecules. In several aspects, a lipid core is stabilized by surfactants and / or emulsifiers as disclosed elsewhere herein, while in other aspects, surfactants are absent. In several aspects, the size of the particle is measured as a mean diameter. In several aspects, the size of the particle can be measured by dynamic light scattering. In several aspects, the size of the particle can be measured using a zeta-sizer. In several aspects, the size of the particle can be measured using Scanning Electron Microscopy (SEM). In several aspects, the size of the particle can be measured using a cryogenic SEM (cryo-SEM). Where the size of a nanoparticle is disclosed elsewhere herein, any one or more of these instruments or methods may be used to measure such sizes.301871278 - 37 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0107] In several aspects, a nanoparticle composition may comprise nanoparticles having an average size of less than or equal to about: 10 nm, 25 nm, 40 nm, 50 nm, 100 nm, 250 nm, 500 nm, 1000 nm, or ranges including and / or spanning the aforementioned values. In several aspects, a composition comprises nanoparticles having an average size of between about 50 nm and 150 nm or between about 50 and about 250 nm. In several aspects, the size distribution of the nanoparticles for at least 50%, 75%, 80%, 90% (or ranges including and / or spanning the aforementioned percentages) of the particles present is equal to or less than about: 20 nm, 40 nm, 60 nm, 80 nm, 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 160 nm, 180 nm, 200 nm, 300 nm, 400 nm, 500 nm, or ranges including and / or spanning the aforementioned nm values. In several aspects, a composition comprises nanoparticles having an average size of less than or equal to about: 10 nm, 50 nm, 100 nm, 250 nm, 500 nm, 1000 nm, or ranges including and / or spanning the aforementioned values. In several aspects, the size distribution of the nanoparticles for at least 90% of the particles present is equal to or less than about: 20 nm, 40 nm, 60 nm, 80 nm, 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 160 nm, 180 nm, 200 nm, 300 nm, 400 nm, 500 nm, or ranges including and / or spanning the aforementioned nm values. In several aspects, the size distribution of the nanoparticles for at least 90% of the particles present is equal to or less than about: 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 160 nm, 180 nm, 200 nm, or ranges including and / or spanning the aforementioned nm values. In several aspects, the D90 of the particles present is equal to or less than about: 80 nm, 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 160 nm, 180 nm, 200 nm, 300 nm, 400 nm, 500 nm, or ranges including and / or spanning the aforementioned values. In several aspects, the size of the nanoparticle is the diameter of the nanoparticle as measured using any of the techniques as disclosed elsewhere herein. For instance, in some aspects, the size of the nanoparticle is the measured using dynamic light scattering. In several aspects, the size of the nanoparticle is the measured using a zeta sizer. In several aspects, consistency in size over time, or within a sample, allows predictable stability for the active agent encapsulated therein.

[0108] In several aspects, over 50%, 75%, 95% (or ranges spanning and or including the aforementioned values) of nanoparticles prepared by methods disclosed herein have a particle size of between about 20 to about 500 nm (e.g., as measured by zeta sizing (e.g., refractive index)). In several aspects, over 50%, 75%, 95% (or ranges spanning and or including the aforementioned values) of nanoparticles prepared by methods disclosed herein have a particle size of between about 50 nm to about 200 nm (e.g., as measured by zeta sizing (e.g., refractive index)). In several aspects, over 50%, 75%, 95% (or ranges spanning and or including the aforementioned values) of nanoparticles prepared by methods disclosed herein have a particle size of between about 90 nm to about 150 nm (e.g., as measured by301871278 - 38 -Atty. Docket Number: UTSDP3820WO- 1001380566zeta sizing (e.g., refractive index)). In several aspects, maintaining consistency in size allows predictable delivery to subjects. In several aspects, the D90 particle size measurement varies between 150 and 500 nm.

[0109] In several aspects, the average size of the nanoparticles of a composition as disclosed herein may be substantially constant and / or does not change significantly over time (e.g., it is a stable nanoparticle). In several aspects, after formulation and storage for a period of at least about 1 month (30 days), about 3 months (90 days), or about 6 months (180 days) (e.g., at ambient conditions, at 25 degrees C with 60% relative humidity, or under the other testing conditions disclosed elsewhere herein), the average size of nanoparticles comprising the composition changes less than or equal to about: 1%, 5%, 10%, 20%, or ranges including and / or spanning the aforementioned values.

[0110] In several aspects, the polydispersity index (PDI) of the nanoparticles of a composition as disclosed herein is less than or equal to about: 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.40, 0.50, 0.60, 0.70, 0.80, or ranges including and / or spanning the aforementioned values. In several aspects, the size distribution of the nanoparticles is highly monodisperse with a polydispersity index of less than or equal to about: 0.05, 0.10, 0.15, 0.20, 0.25, or ranges including and / or spanning the aforementioned values.

[0111] In several aspects, the zeta potential of the nanoparticles of a composition as disclosed herein is less than or equal to about: 1 mV, 3 mV, 4 mV, 5 mV, 6 mV, 7 mV, 8 mV, 10 mV, 20 mV, or ranges including and / or spanning the aforementioned values. In several aspects, the zeta potential of the nanoparticles is greater than or equal to about: -3 mV, -1 mV, 0 mV, 1 mV, 3 mV, 4 mV, 5 mV, 6 mV, 7 mV, 8 mV, 4 mV, 10 mV, 20 mV, or ranges including and / or spanning the aforementioned values. In several aspects, the zeta potential and / or diameter of the particles (e.g., measured using dynamic light scattering) is acquired using a zetasizer (e.g., a Malvern ZS90 or similar instrument).

[0112] In several aspects, as disclosed elsewhere herein, a nanoparticle composition is an oil-in-water emulsion, water-in-oil emulsion, water-in-oil-in-water emulsion, oil-in-water-in-oil emulsion, liposome, solid lipid particles formulation, etc. For brevity, these may just be referred to as the composition. In several aspects, a nanoparticle composition can be processed to comprises one or more of solid lipid nanoparticles, liposomes (and variants including multi-lamellar, double liposome preparations, etc.), niosomes, ethosomes, electrostatic particulates, microemulsions, nanoemulsions, microsuspensions, nanosuspensions, or combinations thereof. In several aspects, polymeric nanoparticles may be formed. In several aspects, cyclodextrin is added.301871278 - 39 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0113] In several aspects, a solid lipid nanoparticle compositions comprises a lipid core matrix. In several aspects, the lipid core matrix is solid. In several aspects, the solid lipid comprises one or more ingredients as disclosed elsewhere herein. In several aspects, the core of the solid lipid comprises one or more lipids, surfactants, active ingredients, etc. In several aspects, the surfactant acts as an emulsifier. In several aspects, emulsifiers can be used to stabilize the lipid dispersion (with respect to charge and molecular weight). In several aspects, the core ingredients (e.g., the components of the core) are present in the composition (collectively or individually) at a dry wt. % of equal to or less than about: 0.5%, 1.0%, 2.5%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 80% or ranges including and / or spanning the aforementioned values. In several aspects, the core ingredients and / or the emulsifiers (collectively or individually) are present in the composition at a wet wt. % of equal to or less than about: 0.5%, 1.0% 2.5%, 5%, 7.5%, 10%, 12.5%, 15%, 20%, 30%, 40%, 60% or ranges including and / or spanning the aforementioned values.

[0114] In several aspects, a nanoparticle composition (e.g., when in water or dried) comprises multilamellar nanoparticle vesicles, unilamellar nanoparticle vesicles, multivesicular nanoparticles, emulsion particles, irregular particles with lamellar structures and bridges, partial emulsion particles, combined lamellar and emulsion particles, and / or combinations thereof. In certain aspects, the nanoparticle compositions do not comprise multilamellar nanoparticle vesicles, unilamellar nanoparticle vesicles, multivesicular nanoparticles, emulsion particles, irregular particles with lamellar structures and bridges, partial emulsion particles, combined lamellar and emulsion particles, and / or combinations thereof. In several aspects, the composition is characterized by having multiple types of particles (e.g., lamellar, emulsion, irregular, etc.). In other aspects, a majority of the particles present are emulsion particles. In several aspects, a majority of the particles present are lamellar (multilamellar and / or unilamellar). In other aspects, a majority of the particles present are irregular particles. In still other aspects, a minority of the particles present are emulsion particles. In several aspects, a minority of the particles present are lamellar (multilamellar and / or unilamellar). In other aspects, a minority of the particles present are irregular particles.

[0115] In several aspects, multilamellar nanoparticles comprise equal to or at least about 5%, 8%, 9%, 10%, 15%, 25%, 50%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in the composition (e.g., the aqueous composition). For example, in some aspects, between about 5% and about 10% of the particles present are multilamellar. In several aspects, unilamellar nanoparticles comprise equal to or at least about 5%, 8%, 9%, 10%, 15%, 20%, 25%, 50%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in301871278 - 40 -Atty. Docket Number: UTSDP3820WO- 1001380566the composition (e.g., the aqueous composition). For example, in some aspects, between about 10% and about 15% of the particles present are unilamellar.

[0116] In several aspects, emulsion particles comprise equal to or at least about 5%, 8%, 9%, 10%, 15%, 25%, 50%, 60%, 65%, 70%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in the composition (e.g., the aqueous composition). For example, in some aspects, between about 60% to about 75% of the particles present are emulsion particles.

[0117] In several aspects, micelle particles comprise equal to or at least about 5%, 8%, 9%, 10%, 15%, 25%, 50%, 60%, 65%, 70%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in the composition (e.g., the aqueous composition). In several aspects, liposomes comprise equal to or at least about 5%, 8%, 9%, 10%, 15%, 25%, 50%, 60%, 65%, 70%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in the composition (e.g., the aqueous composition). In several aspects, irregular particles (including particles with lamellar structures and / or bridges) comprise equal to or at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 25%, 50%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in the composition (e.g., the aqueous composition). For example, in some aspects, between about 1% to about 5% of the particles present are irregular particles. In several aspects, combined lamellar and emulsion particles comprise equal to or at least about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 25%, 50%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in the composition (e.g., the aqueous composition). In several aspects, mixed-micelle particles comprise equal to or at least about 5%, 6%, 7%, 8%, 9%, 10%, 15%, 25%, 50%, 75%, 85%, 95%, or 100% (or ranges spanning and / or including the aforementioned values) of the particles present in the composition (e.g., the aqueous composition).

[0118] The nanoparticle compositions can comprise, but are not limited to, combinations of multilamellar nanoparticles, unilamellar nanoparticles, emulsion nanoparticles, micelle nanoparticles, irregular particles, and / or liposomes.

[0119] The percentages and / or concentrations of particles present in the composition may be purposefully modified. In some aspects, the percentage and / or concentration of the particles present in the composition are tailored to the active compound and / or the liquid comprising the particles. Such tailoring may lead to more homogenization and / or dispersion in the liquid. The tailoring may stabilize dispersion in the liquid.301871278 - 41 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0120] In several aspects, the formulations and / or compositions disclosed herein are stable during sterilization. In several aspects, the sterilization may include one or more of ozonation, LIV treatment, and / or heat treatment. In several aspects, the particle size and / or PDI after sterilization (e.g., exposure to techniques that allow sterilization of the composition) varies by less than or equal to about: 1%, 5%, 10%, 20%, 30%, or ranges including and / or spanning the aforementioned values. In several aspects, the therapeutic agent concentration after sterilization (e.g., exposure to techniques that allow sterilization of the composition) varies (e.g., drops) by less than or equal to about: 1%, 5%, 10%, 15%, or ranges including and / or spanning the aforementioned values.

[0121] In several aspects, the nanoparticle compositions (including after stabilization) disclosed herein have a shelf life of equal to or greater than 1 month, 3 months, 6 months, 12 months, 14 months, 16 months, 18 months, 19 months, or ranges including and / or spanning the aforementioned values. The shelf-life can be determined as the period of time in which there is 95% confidence that at least 50% of the response (active agent(s) concentration or particle size) is within the specification limit. This refers to a 95% confidence interval and when linear regression predicts that at least 50% of the response is within the set specification limit.VI. Dosage formulations

[0122] In certain aspects, the present disclosure provides compositions comprising one or more inhibitors disclosed herein, formulated for one or more routes of administration. Suitable routes of administration may, for example, include intravenous, intramuscular, intraperitoneal, oral, subcutaneous, intranasal route, cranial, transmucosal, trans-nasal, intestinal, and / or parenteral delivery. In some aspects, compositions herein formulated can be formulated for parenteral delivery, specifically intravenous delivery. In some aspects, compositions can be formulated for intramuscular, subcutaneous, intramedullary, intravenous, intraperitoneal, and / or intranasal injections.

[0123] In certain aspects, one may administer a composition herein in a local or systemic manner, for example, via local injection of the pharmaceutical composition directly into a tissue region of a patient. In some aspects, a pharmaceutical composition disclosed herein can be administered parenterally, e.g., by intravenous injection, intracerebroventricular injection, intra-cisterna magna injection, intra-parenchymal injection, or a combination thereof. In some aspects, a pharmaceutical composition disclosed herein can administered to subject as disclosed herein. In some aspects, a pharmaceutical composition disclosed herein can administered to human patient. In some aspects, a pharmaceutical composition disclosed herein can administered to a human patient via at least two administration routes.301871278 - 42 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0124] In certain aspects, pharmaceutical compositions of the present disclosure may be manufactured by processes well known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.

[0125] In certain aspects, pharmaceutical compositions for use in accordance with the present disclosure, thus may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active ingredients into preparations which, can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. For injection, the active ingredients of a pharmaceutical composition herein may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank’s solution, Ringer’s solution, physiological salt buffer, or any combination thereof.

[0126] In certain aspects, pharmaceutical compositions described herein may be formulated for parenteral administration, e.g., by bolus injection or continuous infusion. Formulations for injection herein may be presented in unit dosage form, e.g., in ampoules or in multidose containers with optionally, an added preservative. In some aspects, compositions herein may be suspensions, solutions or emulsions in oily or aqueous vehicles, and / or may contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0127] In certain aspects, pharmaceutical compositions herein formulated for parenteral administration may include aqueous solutions of the active preparation (e.g., a polynucleotide construct) in water-soluble form. In some aspects, compositions herein comprising suspensions of the active preparation may be prepared as oily or water-based injection suspensions. Suitable lipophilic solvents and / or vehicles for use herein may include, but are not limited to, fatty oils such as sesame oil, or synthetic fatty acids esters such as ethyl oleate, triglycerides or liposomes. In some aspects, compositions herein comprising aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, and / or dextran. In some aspects, compositions herein comprising a suspension may also contain one or more suitable stabilizers and / or agents which increase the solubility of the active ingredients (e.g., a polynucleotide construct / vector molecule) to allow for the preparation of highly concentrated solutions.

[0128] In some aspects, compositions herein may comprise the active ingredient in a powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water-based solution, before use.301871278 - 43 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0129] Pharmaceutical compositions suitable for use in context of the present disclosure may include compositions wherein the active ingredients can be contained in an amount effective to achieve the intended purpose. In some aspects, a therapeutically effective amount means an amount of active ingredients (e.g., a polynucleotide construct molecule) effective to prevent, slow, alleviate or ameliorate symptoms of a disorder (e.g., EDE) or prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. For any preparation used in the methods of the present disclosure, the therapeutically effective amount or dose can be estimated initially from in vitro and cell culture assays and or screening platforms disclosed herein. For example, a dose can be formulated in animal models to achieve a desired concentration or titer. Such information can be used to more accurately determine useful doses in humans.

[0130] In some aspects, toxicity and therapeutic efficacy of the active ingredients disclosed herein (e.g., a polynucleotide construct / vector) can be determined by standard pharmaceutical procedures in vitro, in cell cultures or experimental animals. In some aspects, data obtained from these in vitro and cell culture assays and animal studies can be used in formulating a range of dosage for use in a human subject. In some aspects, a dosage for use herein may vary depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient’s condition.

[0131] In certain aspects, dosage amounts and / or dosing intervals may be adjusted individually to brain or blood levels of the active ingredient that are sufficient to induce or suppress the biological effect (minimal effective concentration, MEC). In some aspects, the MEC for an active ingredient (e.g., a polynucleotide construct / vector molecule or composition disclosed herein) may vary for each preparation but can be estimated from in vitro data. In some aspects, dosages necessary to achieve the MEC herein may depend on individual characteristics and route of administration. Detection assays can be used to determine plasma concentrations.

[0132] In certain aspects, depending on the severity and responsiveness of the condition to be treated, dosing with compositions herein can be of a single or a plurality of administrations, with course of treatment lasting from several days to several weeks or until cure is effected or diminution of the disease state is achieved.

[0133] In certain aspects, amounts of a composition herein to be administered will be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, and the like. In some aspects,301871278 - 44 -Atty. Docket Number: UTSDP3820WO- 1001380566effective doses may be extrapolated from dose-responsive curves derived from in vitro or in vivo test systems.

[0134] The active ingredient may be administered at once or may be divided into a number of smaller doses to be administered at intervals of time. The precise dosage and duration of treatment is a function of the disease being treated and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test data. Concentrations and dosage values may also vary with the severity of the condition to be alleviated. For any particular subject, specific dosage regimens can be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.VII. Methods of treatment

[0135] GHSR (growth hormone secretagogue receptor, the ghrelin receptor) is a G-protein-coupled receptor (GPCR) that primarily binds to ghrelin, a hunger-stimulating hormone. GHSR is widely expressed in the several brain regions, pituitary gland, pancreatic islets of Langerhans, gastrointestinal tract, and other tissues, playing a crucial role in regulating metabolism, energy homeostasis, and appetite control. The GHSR binds ghrelin.

[0136] Ghrelin is primarily produced by a subset of gastric and duodenal enteroendocrine cells in adults. Beyond its role as a growth hormone (GH) secretagogue, ghrelin also helps respond to changes in the metabolic state. Its plasma levels increase during caloric restriction and shows an inverse relationship with body weight and adiposity. Administration of ghrelin or agonists of its receptor, the growth hormone secretagogue receptor (GHSR), has been shown to enhance food intake and reward-driven eating behaviors, reduce energy expenditure, and promote weight gain. Conversely, research indicates that inhibiting ghrelin activity can lead to reduced baseline food intake, diminished refeeding after fasting, lower food consumption following exercise, decreased hedonic eating, and prevention of weight gain and fat accumulation.

[0137] Upon ghrelin binding, GHSR activation triggers intracellular signaling pathways, including the cAMP / PKA and PI3K / Akt pathways, which influence appetite, growth hormone secretion, and insulin sensitivity. GHSR is a key regulator of energy balance and metabolic processes. Activation of the receptor stimulates appetite by promoting the release of neuropeptide Y (NPY) and agouti-related peptide (AgRP) in the hypothalamus, leading to increased food intake. Additionally, GHSR influences glucose metabolism by modulating insulin secretion and sensitivity, as well as promoting lipogenesis and inhibiting fat oxidation. In the gastrointestinal tract, ghrelin signaling affects gastric motility, acid secretion, and gut-301871278 - 45 -Atty. Docket Number: UTSDP3820WO- 1001380566brain communication, further contributing to metabolic regulation. Dysfunction of the ghrelin receptor has been implicated in several metabolic and neuropsychiatric disorders. Conditions like eating disorders, cachexia, Prader-Willi Syndrome, and short stature, have been associated with altered ghrelin levels and / or ghrelin signaling, contributing to severe weight loss and metabolic imbalances. Moreover, its role in reward processing and stress response links it to neuropsychiatric disorders such as depression, anxiety, and addiction. Emerging research also suggests that GHSR may play a role in neurodegenerative diseases, including Parkinson’s disease, amyotrophic lateral sclerosis or other neurodegenerative disorders, and aging-related memory decline, where ghrelin signaling may have neuroprotective effects.

[0138] LEAP2 (Liver-expressed antimicrobial peptide 2) is a metabolic hormone, produced mainly by the liver and small intestine, which serves as an antagonist and inverse agonist to GHSR. By binding to GHSR, LEAP2 inhibits both the constitutive (ligand-independent) activity of the receptor and the activity induced by ghrelin. This antagonistic action effectively suppresses the orexigenic (appetite-stimulating) and growth hormone-releasing effects typically mediated by ghrelin. LEAP2 reduces ghrelin-induced food intake and GH secretion and is found elevated in obesity but lowered upon fasting. Thus, the plasma LEAP2 / ghrelin molar ratio could be a key determinant modulating GHSR signaling in response to changes in body mass and feeding status. Additionally, studies have shown that LEAP2 analogs may improve hepatic steatosis, lobular inflammation, fibrosis, notable reductions in body weight and plasma total cholesterol levels, suggesting LEAP2's therapeutic potential in addressing metabolic-associated fatty liver disease (MAFLD) and associated metabolic disorders. Conversely, presence of LEAP2 can negatively impact ghrelin associated physiological functions. Without being bound by theory, reducing the levels of LEAP2 can therefore positively impact GHSR and ghrelin related signaling and related functions. LEAP2 deletion in mice enhances sensitivity to ghrelin, resulting in increased food intake and growth hormone secretion. In female mice, the absence of LEAP2 leads to increased body length and exacerbates hepatic fat accumulation when subjected to a high-fat diet, highlighting LEAP2's role in growth regulation and lipid metabolism.

[0139] Therefore, in some aspects, the current disclosure encompasses a method of treating a disease or a disorder, in a subject in need thereof, by reducing the levels of LEAP2, and / or enhancing GHSR function, for example ghrelin related signaling. In some aspects, the method comprises administering to the subject one or more of the anti-LEAP2 aptamer disclosed herein. In some aspects, the subject has or is suspected of having a ghrelin and / or GHSR function linked disease or disorder. Ghrelin and GHSR-linked diseases or disorders involve dysregulation of ghrelin signaling, leading to abnormalities in appetite control, metabolism, and hormone regulation. In particular, diseases and / or disorders that can be treated using the301871278 - 46 -Atty. Docket Number: UTSDP3820WO- 1001380566methods of the current disclosure may correspond to those caused by inhibition or reduced functioning of the ghrelin receptor (GHSR), characterized by impaired appetite regulation, growth hormone secretion, and metabolic balance. Conditions such as cachexia, growth hormone deficiency, and gastroparesis can arise when GHSR signaling is diminished, leading to loss of appetite, unintended weight loss, reduced energy intake, and slowed gastric motility. In cases like cachexia associated with cancer or chronic illness, ghrelin resistance may limit the body’s attempts to minimized muscle wasting and metabolic decline. In some aspects, the ghrelin and / or GHSR linked disease, or disorder is cachexia. Thus, in some aspects, the current disclosure encompasses of treating, preventing or minimizing cachexia in a subject comprising administering to a subject in need thereof a pharmaceutical composition or compositions comprising one or more anti-LEAP2 aptamers disclosed herein. In some aspects, the cachexia may be associated with cancer, or a treatment for cancer. Certain anticancer agents, such as cisplatin, have one or more undesirable adverse effects that involve causing or increasing one or more syndromes such as cachexia, sarcopenia, muscle wasting, bone wasting or involuntary body weight loss. Accordingly, in certain aspects, the present disclosure comprises a method of treating cancer, while preventing, minimizing or reducing the occurrence, frequency or severity of cachexia, sarcopenia, or muscle wasting, bone wasting or involuntary loss of body weight in a subject, comprising administering to the subject in need thereof a pharmaceutical composition comprising one or more anti-LEAP2 aptamers. In some aspects, the subject may be administered a combination therapeutic comprising one or more anti-LEAP2 aptamers in combination with one or more anti-LEAP2 shRNAs, one or more GHSR agonists or any combination thereof as disclosed herein below. Other forms of cachexia, including, but not limited to cardiac cachexia, pulmonary cachexia, renal cachexia, or cachexia of aging can also be treated by a method disclosed herein.

[0140] In some aspects, the current disclosure also encompasses treating of other ghrelin and / or GHSR signaling related diseases or disorders. Non-limiting examples include sarcopenia, anorexia nervosa, bulimia nervosa, eating disorders, hypoglycemia, depression, anxiety, Alzheimer’s disease, dementia, age-related memory decline, gastrointestinal dysmotility disorders (postoperative ileus, diabetic gastroparesis), hypoglycemia unawareness, post-traumatic stress disorder, amyotrophic lateral sclerosis or other neurodegenerative disorders, type 1 diabetes, type 2 diabetes, obesity, or short stature. Additionally, emesis such as caused by cancer chemotherapy, constipation such as associated with the hypomotility phase of irritable bowel syndrome (IBS), delayed gastric emptying associated with wasting conditions, gastroesophageal reflux disease (GERD), gastric ulcers may also be treated using one or more of the compositions disclosed herein. In some aspects, the LEAP2 aptamers enhance selective activation of the ghrelin receptor to301871278 - 47 -Atty. Docket Number: UTSDP3820WO- 1001380566provide an avenue for the treatment of diseases characterized by gastrointestinal (Gl) dysmotility. In some aspects, surgical manipulation of the stomach or intestine causes a disorganization of the gut-brain signaling pathways, impairing Gl activity. GHSR activation stimulate and coordinate the firing of vagal afferent neurons and thereby modulate gut motility, thus accelerating gastric emptying. In some aspects, administration of the anti-LEAP2 aptamers, or compositions thereof, can enhance GHSR signaling, and hence can be used for treatment of any of the aforementioned conditions. In some aspects, administration of the anti-LEAP2 aptamers, or compositions thereof, can enhance ghrelin activity by reducing antagonism to GHSR function, thereby improving coordinated firing of vagal afferent neurons and thereby gut motility. Gastroparesis (“stomach paralysis”) is a syndrome characterized by delayed gastric emptying in the absence of any mechanical obstruction. It is variably characterized by abdominal pain, nausea, vomiting, weight loss, anorexia, early satiety, malnutrition, dehydration, gastroesophageal reflux, cramping and bloating. This chronic condition can lead to frequent hospitalization, increased disability and decreased quality of life. Severe, symptomatic gastroparesis is common in individuals suffering from diabetes, affecting from 5-10% of diabetics for a total patient population of 1 million in the U.S. alone. Neuropathy is a frequent, debilitating complication of diabetes. Visceral neuropathy results in Gl dysfunction, especially involving the stomach, leading to impaired gastric motility. Enhancing ghrelin function and / or GHSR function by decreasing antagonism to the GHSR receptor would therefore be highly effective in overcoming the fundamental motility barrier faced by gastroparesis patients and correcting this condition. Opioid-induced bowel dysfunction is the term applied to the confluence of symptoms involving the reduced Gl motility that results from treatment with opioid analgesics. Approximately 40-50% of patients taking opioids for pain control experience OBD. It is characterized by hard, dry stools, straining, incomplete evacuation, bloating, abdominal distension and increased gastric reflux. In addition to the obvious short-term distress, this condition leads to physical and psychological deterioration in patients undergoing long term opioid treatment. Further, the dysfunction can be so severe as to become a dose-limiting adverse effect that actually prevents adequate pain control. In some aspects, the anti-LEAP2 aptamers may be useful to alleviate symptoms, slow disease progression, and improve the quality of life for subjects suffering from any one or more of the diseases or disorders disclosed herein above.

[0141] In some aspects, administration of the aptamer into a subject in need thereof, may reduce the level of functional LEAP2 peptide by at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% compared to baseline, wherein the baseline corresponds to the level of LEAP2 peptide in the subject prior to administration. In certain aspects, treatments administered according to the methods disclosed herein reduce301871278 - 48 -Atty. Docket Number: UTSDP3820WO- 1001380566symptoms and / or can improve patient life expectancy compared to the life expectancy and / or symptoms an untreated subject with identical disease condition and predicted outcome. As used herein, “patient life expectancy” is defined as the time at which 50 percent of subjects are alive and 50 percent have passed away. In some aspects, patient life expectancy can be indefinite following treatment according to the methods disclosed herein. In other aspects, patient life expectancy can be increased at least about 5% or greater to at least about 100%, at least about 10% or greater to at least about 95% or greater, at least about 20% or greater to at least about 80% or greater, at least about 40% or greater to at least about 60% or greater compared to an untreated subject with identical disease condition and predicted outcome. In some aspects, patient life expectancy can be increased at least about 5% or greater, at least about 10% or greater, at least about 15% or greater, at least about 20% or greater, at least about 25% or greater, at least about 30% or greater, at least about 35% or greater, at least about 40% or greater, at least about 45% or greater, at least about 50% or greater, at least about 55% or greater, at least about 60% or greater, at least about 65% or greater, at least about 70% or greater, at least about 75% or greater, at least about 80% or greater, at least about 85% or greater, at least about 90% or greater, at least about 95% or greater, at least about 100% compared to an untreated subject with identical disease condition and predicted outcome. In some aspects, patient life expectancy can be increased at least about 5% or greater to at least about 10% or greater, at least about 10% or greater to at least about 15% or greater, at least about 15% or greater to at least about 20% or greater, at least about 20% or greater to at least about 25% or greater, at least about 25% or greater to at least about 30% or greater, at least about 30% or greater to at least about 35% or greater, at least about 35% or greater to at least about 40% or greater, at least about 40% or greater to at least about 45% or greater, at least about 45% or greater to at least about 50% or greater, at least about 50% or greater to at least about 55% or greater, at least about 55% or greater to at least about 60% or greater, at least about 60% or greater to at least about 65% or greater, at least about 65% or greater to at least about 70% or greater, at least about 70% or greater to at least about 75% or greater, at least about 75% or greater to at least about 80% or greater, at least about 80% or greater to at least about 85% or greater, at least about 85% or greater to at least about 90% or greater, at least about 90% or greater to at least about 95% or greater, at least about 95% or greater to at least about 100% compared to an untreated patient with identical disease condition and predicted outcome.

[0142] As used herein, “an effective amount” refers to the amount of each active agent required to confer therapeutic effect on the subject, either alone or in combination with one or more other active agents. An effective amount may refer to a dose of compound sufficient to provide circulating or local concentrations high enough to impart a beneficial effect on the301871278 - 49 -Atty. Docket Number: UTSDP3820WO- 1001380566recipient thereof. The precise amount to be administered can be determined by the skilled practitioner in view of desired dosages, side effects, and medical history of the patient. Determination of whether an amount of the nucleic acids, the small molecules, or the polypeptides / peptide, disclosed herein has achieved a therapeutic effect would be evident to one of skill in the art. Effective amounts vary, as recognized by those skilled in the art, depending on the condition being treated, the severity of the condition, the individual patient parameters including age, physical condition, size, gender and weight, the duration of the treatment, the nature of concurrent therapy (if any), the specific route of administration and like factors within the knowledge and expertise of the health practitioner. These factors are well known to those of ordinary skill in the art and can be addressed with no more than routine experimentation. In some aspects, a maximum dose of the individual components or combinations thereof may be used, that is, the highest safe dose according to sound medical judgment. In some aspects, empirical considerations, such as the half-life, generally will contribute to the determination of the dosage. In some aspects, frequency of administration may be determined and adjusted over the course of therapy, and is generally, but not necessarily, based on treatment and / or suppression and / or amelioration and / or delay of a target disease / disorder. In some aspects, sustained continuous release formulations of an antibody herein may be appropriate. Various formulations and devices for achieving sustained release are known in the art and provided herein.

[0143] In some aspects, dosages as described herein may be determined empirically in individuals who have been given one or more administration(s) of an activator disclosed herein. In accordance with some aspects herein, individuals can be given incremental dosages. To assess efficacy of the antibody, polynucleotide or small molecule, an indicator of the disease / disorder can be followed.

[0144] In some aspects, for administration of any of the pharmaceutical compositions described herein, an initial candidate dosage can be about 2-200 mg / kg. For the purpose of the present disclosure, a typical daily dosage may range from about any of 0.1 pg / kg to 3 pg / kg to 30 pg / kg to 300 pg / kg to 3 mg / kg, to 30 mg / kg to 300 mg / kg weight of the subject or more, depending on the factors mentioned above. In some aspects, a typical daily dosage may be 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg, 34 mg / kg, 35 mg / kg, 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg, 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg weight of the subject. In some aspects, for an adult patient of normal weight, doses ranging from about 0.5 to about 200 mg / kg may be administered. In some301871278 - 50 -Atty. Docket Number: UTSDP3820WO- 1001380566aspects, the dosage described herein can be less than, more than, equal to or about 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg, 34 mg / kg, 35 mg / kg, 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg, 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg, or more. The particular dosage regimen, i.e., dose, timing and repetition, can depend on the particular individual and that individual’s medical history, as well as the properties of the individual agents (such as the half-life of the agent, and other considerations well known in the art). For repeated administrations over several days or longer, depending on the condition, treatment methods of the present disclosure may be sustained until a desired suppression of symptoms occurs and / or until sufficient therapeutic levels are achieved to alleviate a target disease or disorder, or a symptom thereof. In some aspects, a dosing regimen herein may comprise administering a dose of about 2-200 mg / kg. In some aspects, other dosage regimens may be useful, depending on the pattern of pharmacokinetic decay that the practitioner wishes to achieve. In accordance with some aspects herein, dosing from one-four times a week is contemplated. In some aspects, dosing ranging from about 0.5 mg / kg to about 200 mg / kg may be used. In some aspects, dosing frequency may be more than, less than, about or equal to twice daily, once daily, twice weekly, once every week, every 2 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or every 10 weeks; or once every month, every 2 months, or every 3 months, or longer. In some aspects, the progress of this therapy can be easily monitored by conventional techniques and assays. In some aspects, a dosing regimen (including the disclosed antibody used) suitable for use herein can vary over time.

[0145] In some aspects, the pharmaceutical compositions disclosed herein may be administered to the subject by a variety of routes. For example, one or more of the compositions disclosed herein may be administered orally via a solid or liquid dosage form (tablet, gel cap, time release capsule powder, solution, or suspension in aqueous or nonaqueous liquid), parenterally (i.e., subcutaneously, intradermally, intravenously, (i.e., as a solution, suspension, or emulsion in a carrier), and / or intramuscularly, intracranially, or intraperitoneally). In one aspect, the compounds may be administered in saline or with a pharmaceutically acceptable excipient as described above.

[0146] In some aspects, a suitable subject, or a subject in need thereof is a subject diagnosed with, showing symptoms of, suspected of having, or having susceptibility to a ghrelin or ghrelin receptor (GHSR) linked disease or disorder. Non-limiting examples include cachexia, anorexia301871278 - 51 -Atty. Docket Number: UTSDP3820WO- 1001380566nervosa, hypoglycemia, depression, anxiety, Alzheimer’s disease, dementia, age related memory decline, gastrointestinal dysmotility disorders (postoperative ileus, diabetic gastroparesis), hypoglycemia unawareness, post-traumatic stress disorder, amyotrophic lateral sclerosis or other neurodegenerative disorders, type 1 diabetes, type 2 diabetes, obesity, or short stature. Suitable subjects may include, without limit, humans, as well as companion animals such as cats, dogs, rodents, and horses; research animals such as rabbits, sheep, pigs, dogs, primates, mice, rats, and other rodents; agricultural animals such as cows, cattle, pigs, goats, sheep, horses, deer, chickens, other fowl; zoo animals; and primates such as chimpanzees, monkeys, and gorillas. The subject can be of any age without limitation. In an aspect, the subject may be a human.

[0147] Generally, the composition will be administered in a therapeutically effective amount which includes prophylactic amounts or lower dosages for example, when combined with another agent. In certain aspects, a compound disclosed herein may be administered to a subject intravenously at a concentration ranging from about 0.5 mg / kg to about 200 mg / kg. In some aspects, a compound disclosed herein may be administered to a subject intravenously at a concentration of about less than, more than, equal to or about 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, about 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg, or more. In some aspects, a compound disclosed herein may be administered to a subject intravenously at least once a day, at least twice a day, at least three times a day, or more.

[0148] In certain aspects, a compound disclosed herein may be administered to a subject orally at a concentration ranging from about 5 mg / kg to about 500 mg / kg. In some aspects, a compound disclosed herein may be administered to a subject orally at a concentration of about less than, more than, equal to or about 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, about 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 150 mg / kg, 200 mg / kg, or more. In some aspects, a compound disclosed herein may be administered to a subject orally at least once a day, at least twice a day, at least three times a day or more.Additional therapies

[0149] In some aspects, the one or more of the pharmaceutical compositions disclosed herein may be administered together with other therapeutic interventions. In some aspects, the compositions disclosed herein can be administered before, after or along with additional therapeutics and treatment methods. In some aspects, the additional therapeutic or treatment method may be administered concurrently. As used herein, a therapy or therapeutic that is administered “concurrently” with another drug is administered within 1 day of the other drug.301871278 - 52 -Atty. Docket Number: UTSDP3820WO- 1001380566In some aspects, a therapy or therapeutic that is administered concurrently with another drug is administered at about the same time, within about 5 minutes, within about 10 minutes, within about 15 minutes, within about 20 minutes, within about 30 minutes, within about 45 minutes, within about 1 hour, within about 2 hours, within about 3 hours, within about 4 hours, within about 5 hours, within about 6 hours, within about 7 hours, within about 8 hours, within about 9 hours, within about 10 hours, within about 11 hours, within about 12 hours, within about 13 hours, within about 14 hours, within about 15 hours, within about 16 hours, within about 17 hours, within about 18 hours, within about 19 hours, within about 20 hours, within about 21 hours, within about 22 hours, within about 23 hours, or within about 24 hours of administration of the other drug. In some aspects, additional therapies or therapeutics aim to alleviate symptoms, slow disease progression, and improve the quality of life for subjects in need thereof, for example a subject diagnosed with, showing symptoms of, suspected of having, or having susceptibility to a ghrelin, and / or GHSR related disease or disorder. These additional therapies may include, but are not limited to pharmacological treatments, non-pharmacological interventions, and emerging experimental therapies.

[0150] In some aspects, the one or more additional therapies may include administration of a shRNA that targets a LEAP2 mRNA. In some aspects, the shRNA induces gene silencing and / or reduction in mRNA levels by targeting complementary sequences within the LEAP2 mRNA, leading to its degradation or translational repression. In some aspects, the shRNA may be formulated with the disclosed anti-LEAP2 aptamers, or separately for administration concurrent to, or staggered with the disclosed aptamers.

[0151] In some aspects, the one or more additional therapies may comprise a GHSR agonist. GHSR (growth hormone secretagogue receptor) analogs are compounds that activate the receptor, influencing processes such as appetite regulation, metabolism, and growth hormone (GH) secretion. These analogs are primarily designed to mimic the effects of ghrelin, the natural ligand of GHSR, and have been explored for various medical applications, including metabolic disorders, growth hormone deficiencies, and gastrointestinal conditions. In some aspects, the GHSR agonist is a ghrelin analog. Ghrelin analogs are synthetic or modified peptides designed to mimic the effects of ghrelin by binding to and activating GHSR. These analogs have been developed primarily for their ability to stimulate appetite, promote growth hormone (GH) secretion, and influence metabolism. Some of the most well-known ghrelin analogs include Macimorelin, Anamorelin, and Relamorelin, each with distinct medical applications.

[0152] Macimorelin is an orally active ghrelin receptor agonist approved by the FDA and EMA for diagnosing adult growth hormone deficiency (AGHD). It stimulates GH release in a manner similar to endogenous ghrelin, allowing for a non-invasive diagnostic alternative to traditional301871278 - 53 -Atty. Docket Number: UTSDP3820WO- 1001380566GH stimulation tests. Anamorelin, another ghrelin analog, has been investigated for its role in treating cancer-associated cachexia. By stimulating appetite and increasing lean body mass, Anamorelin has shown promise in improving the quality of life for patients suffering from severe weight loss due to cancer. Relamorelin, a synthetic ghrelin analog, has been studied for its potential in treating gastroparesis, a condition characterized by delayed gastric emptying often seen in diabetes. It enhances gastrointestinal motility by mimicking ghrelin’s effects on gut function, making it a potential therapeutic option for patients with gastrointestinal disorders. These ghrelin analogs highlight the broad therapeutic potential of ghrelin receptor agonists in treating metabolic, endocrine, and gastrointestinal conditions.EXAMPLES

[0153] LEAP2 (Liver-expressed antimicrobial peptide 2) is a metabolic hormone, produced mainly by the liver and small intestine, which serves as an antagonist and inverse agonist to the ghrelin receptor (growth hormone secretagogue receptor (GHSR)). In some aspects, the current disclosure is based on the discovery that activity of GHSR agonists may be limited by the presence of LEAP2. Decreasing LEAP2 levels with anti-LEAP2 aptamer(s) reduces ghrelin resistance and enhances the activity of endogenous ghrelin, ghrelin-independent GHSR activity, and the activity of administered GHSR agonists, thus providing effective treatment options against diseases like cachexia. Additionally, it is also discussed herein, that the therapeutic benefits of anti-LEAP2 aptamer(s) may be further enhanced by combining these therapeutics with additional therapies, including one or more anti-LEAP2 shRNAs, and / or GHSR agonists. Presented here is data showing the development, selection, and in vitro and in vivo testing of these aptamers.Example 1 : Library Selection and SELEX (Systematic Evolution of Ligands by Exponential enrichment)

[0154] In order to design effective aptamers, a library of aptamers were generated and subjected to SELEX (Systematic Evolution of Ligands by Exponential enrichment). A plate based selection method was utilized where a commercially-available anti-LEAP2 antibody (Phoenix Pharmaceuticals) was coated onto the walls of the plate wells (wells with a “negative protein mix”). A separate set of plates were made by coating the walls of the plate wells with anti-LEAP2 antibody followed by the addition of LEAP2. The aptamer pool was incubated in wells containing the negative protein mix to deplete non-specific binding DNAs before being transferred to a well containing the anti-LEAP2 antibody and LEAP2 protein. After incubation the supernatant was discarded, the wells washed, and the bound aptamers eluted for PCR and single strand DNA (ssDNA) generation. The ssDNA generated from each round was301871278 - 54 -Atty. Docket Number: UTSDP3820WO- 1001380566carried forward into the subsequent round and monitored for binding using biotinylated aptamer and streptavidin-HRP mediated colorimetric readouts.

[0155] During selections the pool showed moderate signal, but also displayed some background binding, especially when used in conjunction with an antibody. Therefore, experiments were conducted reverting to an earlier round and reselection was started with a more stringent selection method. After reselection an aptamer library was generated that had a high likelihood of having specific aptamers for the LEAP2 protein. Before sending the library for sequencing a final binding evaluation was performed to ensure at least 2x signal-to-noise ratio (SNR). The final test is shown below in Table 1.Table 1 : Test results for LEAP2 aptamer libraryExample 2: Next-Generation Sequencing (NGS) and Aptamer Family Characterization

[0156] Aptamer library was sequenced using NGS technology. Libraries were constructed by using a common set of 5’ and 3’ primers, each of which is a 30-mer and is incorporated into the aptamer sequences. Once the NGS data was received the data was analyzed and it was discovered that a single sequence represented approximately 16% of the total reads and was thus the most common sequence. This is a very high percentage of the population, with single sequences in the low single digit percentage ranges generally being considered “high”. There were 3 other sequences of particular note, a sequence that had 2 occurrences in the top 10 sequences, varying only by a single base mutation, which represented 1.8% of the total population. A third sequence also represented 0.5% of the total population. There was a significant 8 base motif that was present throughout the library, “GCTACGTG”, which represented over 1% of the total population. This motif tended to be toward the 3’ end of the random region, with little appreciable pattern in regards to the remaining random sequence. A total of 10 sequences, which included the top 8 most populous sequences (the two sequences that varied by only a single base, only the more populous one was used) as well as 2 sequences that contained the “GTACGTG” motif were used for testing.301871278 - 55 -Atty. Docket Number: UTSDP3820WO- 1001380566Example 3: Candidate testing (binding affinity)

[0157] The initial test of the sequences showed that all of the selected sequences showed positive binding for LEAP2 over the negative control. For all tests below the aptamer was folded by heating to 95 °C for 10 minutes before cooling on an ice bath. The folding buffer used was 5 mM KCI, 20 mM NaCI, 1 mM CaCh, 100 JJM MgCh, 20 mM Tris pH 7.6. Results are shown in Table 2.Table 2: Binding data

[0158] Based on the binding assay four candidate aptamers were identified (SEQ ID NOS: 7-10). In the sequences below the sequence that is unique to each aptamer is underlined, while the remaining sequence corresponds to the 5’ and 3’ primer sequences.Aptamer 2 5’CACACACTGTCCTCGCTCTATTTTCTTCACGCAGAAAAAGCACTCCAGGTATGGGAGA GCGTGGGACTTGACATCTGACTATTACGACTG -3’ [SEQ ID NO: 7]Aptamer 3:5’CACACACTGTCCTCGCTCTATTTTCTTCACGATGGCCCCTTGTGGTCCCTGGCTCGGT GGGACTTGACATCTGACTATTACGACTG-3’ [SEQ ID NO: 8]Aptamer 7:5’CACACACTGTCCTCGCTCTATTTTCTTCACTGGTCACGATTCGGAGTTGGCTGGTTAT GGGTGGGACTTGACATCTGACTATTACGACTG-3 [SEQ ID NO: 9]Aptamer 8:5’CACACACTGTCCTCGCTCTATTTTCTTCACCACTATTTGTTAACTCGGATAGGCTACGT GGTGGGACTTGACATCTGACTATTACGACTG-3’ [SEQ ID NO: 10]

[0159] Interestingly, sequence 8, which was one of the two sequences that contained the “GCTACGTG” motif had the highest signal. However, upon repeat testing the high signal could not be replicated, and sequence 7 emerged as a strong candidate. The 8-mer sequence in Aptamer 8 that is in bold corresponds to a motif enriched in many (1%) of the anti-LEAP2 aptamer candidates identified in an early screen.301871278 - 56 -Atty. Docket Number: UTSDP3820WO- 1001380566

[0160] The selected candidate aptamers were synthesized and tested with 4 distinct modifications each. (These modifications were made using standard phosphonothioate chemistry):1. 5’-biotinylated version [with a biotin group at the 5’-end]2. 5’-PEGylated version [with 3 ethylene oxides: H-(O-CH2-CH2)3-OH - or rather, a triethylene glycol spacer - at the 5’-end]3. 3’-PEGylated version [with 3 ethylene oxides: H-(O-CH2-CH2)3-OH - or rather, a triethylene glycol spacer - at the 3’-end]4. 5’+3’-PEGylated version [with 3 ethylene oxides: H-(O-CH2-CH2)3-OH- or rather, a triethylene glycol spacer - at both the 5’-end and 3’-end]

[0161] Aptamer 7 was used as a candidate for subsequent studies. Aptamer dilution showed that increasing the aptamer concentration of sequence 7 (aptamer 7) increased the signal given by the aptamer, but did not increase the background binding. Using 5 ug / mL of coated LEAP2 antigen, binding using as little as 800 pM aptamer was obtained (see Table 3), although based on the results higher amounts of aptamer appears to give much stronger signal without an increase in background. This highlights the specificity of the aptamer binding to the antigen.Table 3: Binding studies with diluted aptamer 7

[0162] It was further tested, if non-specific DNA could inhibit recognition of the target antigen. LEAP2 antigen coated plate were blocked with 1 pg / mL of salmon sperm DNA, and the DNA was also added to the aptamer incubation mix to determine if the non-specific DNA would301871278 - 57 -Atty. Docket Number: UTSDP3820WO- 1001380566interfere with binding. No degradation in the signal was observed when exogenous DNA was added to the assay (see Table 4).Table 4: Non-specific DNA could not compete-out the binding of the aptamer.

[0163] Finally, the limit of detection (LOD) of the aptamer was tested to detect plate bound LEAP2 antigen. The aptamer is able to detect antigen down to approximately 312 ng / mL (see Table 5).Table 5: Limit-of-detection of antigen (ag) by the LEAP2 aptamer 7Example 4: Cell based assays

[0164] Cell based assays were conducted to confirm that the aptamers could enhance ghrelin-dependent GHSR signaling which otherwise is reduced by LEAP2. GHSR-Tango p-arrestin recruitment assay was used as elaborated below.

[0165] GHSR-Tango P-arrestin recruitment assay: GHSR activity was examined by the Tango p-arrestin recruitment assay using HTLA cells that stably express tTA-dependent luciferase reporter and p-arrestin2-TEV fusion gene. Experimentally, HTL cells were maintained in DMEM (Corning) containing 10% FBS (Thermo Fisher), 100 ll / ml penicillin, 100|jg / ml streptomycin (Gibco), 2 pg / ml puromycin (Tocris Bioscience), and 100 pg / ml hygromycin B301871278 - 58 -Atty. Docket Number: UTSDP3820WO- 1001380566(Corning) at 37 °C with 5% CO2. Cells were passaged to poly-D-lysine-coated 96-well clear bottom plate (Costar 3610) at 2 x 104per well and maintained for 24 hours before transfection. Thereafter, HTLA cells were transfected with GHSR-Tango plasmid expressing human GHSR and a V2 tail (Addgene 66293, 100ng / well) using calcium phosphate transfection and maintained for 24 hours prior to the treatment. The ECso of human ghrelin peptide (Phoenix Pharmaceuticals, #031-30) was determined using varying doses of tested compounds. To determine the ICso of LEAP2 peptide (Phoenix Pharmaceuticals, #075-40), GHSR-Tango transfected cells were pre-treated with serial doses of LEAP2 peptide for 2 hours followed by human ghrelin peptide treatment at its ECso. T o establish the ECso of LEAP2 aptamers, GHSR-Tango transfected cells were treated with LEAP2 peptide at its predetermined ICso and serial doses of LEAP2 aptamers for two hours followed by human ghrelin peptide treatment at its ECso- To examine the effect of LEAP2 aptamers, GHSR-Tango transfected HTLA cells were exposed to different doses of LEAP2 peptide in the presence or absence of LEAP2 aptamers at their ECso for 2 hours followed by the coincubation with human ghrelin peptide at its ECso for another 16-18 hours. The cells were then incubated with Bright-Glo Luciferase Assay solution (Promega #E2610), and the intensity of luciferase was measured on a Biotek Neo2 microplate reader. Data were analyzed using GraphPad Prism 9 software. The nonlinear regression model of log(agonist) or log(antagonist) vs response were employed to calculate the ECso of human ghrelin peptide and LEAP2 aptamers, or ICso of LEAP2 peptide. The competitive enzyme kinetics equation was used to determine the Ki of LEAP2, with and without anti-LEAP2 aptamers.

[0166] To investigate the efficacy of LEAP2 inhibition using DNA aptamers, libraries were screened for single-strand DNA aptamers that would bind and neutralize circulating LEAP2. Of the top ten hits with the strongest and most specific binding affinity to plate-bound LEAP2, aptamers 2, 3, 7, and 8 were tested. In view of the capacity of LEAP2 to block ligand-mediated GHSR activation by competing with ghrelin, the selected aptamers were tested for their ability to obstruct LEAP2 function with a ghrelin-induced p-arrestin recruitment assay. To examine the effect of the four aforementioned anti-LEAP2 aptamers 2, 3, 7, and 8, GHSR-Tango-expressing HELA cells were treated with 5’-biotinylated versions of aptamers at doses from 1 nM, 10 nM, and 100 nM in the presence of both 10 nM ghrelin and 10 nM LEAP2. Vehicle treatment without aptamer and LEAP2 was used as a critical positive control for ghrelin-induced GHSR activation. Analysis of the results indicate that among the 4 tested aptamers, anti-LEAP2 aptamer 7 had the greatest efficacy against LEAP2’s effect on GHSR activation (FIG. 1A). In contrast, anti-LEAP2 aptamer 8 demonstrated no effect (FIG. 1A) and was subsequently chosen as a negative control for future planned studies. Further tests were done to determine which of the 4 different modified versions of aptamer 7 worked best. The 5’-301871278 - 59 -Atty. Docket Number: UTSDP3820WO- 1001380566biotinylated version and the 5’- PEGylated version best protected ghrelin-mediated GHSR activation from LEAP2 (FIG. 1B), followed by the 3’+5’-PEGylated version and the 3’-PEGylated version (FIG. 1B). Next, the ECso of the 5’-biotinylated version of Aptamer 7 was determined to be 1.26 nM (FIG. 1C). Next, the effect of the 5’-biotinylated aptamer 7 on the ability of LEAP2 to inhibit ghrelin-dependent GHSR activity was tested. Dose-response curves were generated using a consistent dose of ghrelin but with increasing concentrations of LEAP2 with or without 5’ biotin-anti-LEAP2 aptamer 7. As shown in FIG. 1D, 5’ biotinylated aptamer 7 raised the IC50 of LEAP2 from 14.58 nM to 74.08 nM with reduced LEAP2 binding affinity to GHSR demonstrated by the Kj shifted from 14.8 nM to 48.71 nM.Example 5: In vivo mouse experiments

[0167] Next, experiments were designed to test if animals (mice) treated with aptamer showed a greater stimulation (P=0.088) in ghrelin-induced growth hormone (GH) release. For this, a quantitative PCR based assay was developed to detect administered biotinylated anti-LEAP2 aptamer 7 in plasma of mice. A serial dilution of aptamer was prepared ranging from 616 ng to 6.16 pg. One pL of each dilution was used in a 10 pL qPCR reaction. The results are seen in FIG. 2. In FIG. 2, the Y axis is the CT value of the qPCR, and the X axis is the log of the actual concentration of diluted aptamer added to the PCR reaction tube. Results show that this newly-designed custom TaqMan Gene expression assay can detect biotinylated anti-LEAP2 aptamer 7 with >95% efficiency and R2 value of 0.9973, and the range of aptamer detection for this assay was found to be: 0.616 pg to 0.00000616 pg.

[0168] Once a detection assay was developed, experiments were conducted to detect administered biotinylated anti-LEAP2 aptamer 7 in plasma following using different modes of administration - intravenous (IV), intraperitoneal (IP), and subcutaneous (SC). Three mice were injected either using IV (via tail vein), IP, or SC administration with aptamer at a dose of 1.4 pg / g bodyweight. This dose is estimated to be 100X the amount of circulating LEAP2 in obese mice. Blood was collected at 15-, 60- , 120-, and 240-min post-injection. Blood was spun down to isolate the plasma and then diluted 1:10 with water. Then, 2 pL of this dilution was used in the qPCR assay. Concentrations were estimated based on the LEAP2 aptamer assay test designed above. FIG. 3 provides the results of the in vivo test. As seen from the figure, there was little difference between the amount of aptamer detected in the mice injected IP vs IV, however in general, a greater amount of aptamer was detected following IV administration than the other methods. There was one mouse in the IV group that had a much larger amount of aptamer detected at all time points. This could be due to a more successful tail vein injection in that single mouse. Due to similar amounts detected in the remaining two mice in the IV group vs. the 3 in the IP group, and considering the enhanced technical feasibility of IP delivery, subsequent sets of in vivo GH and food intake experiments were performed using IP301871278 - 60 -Atty. Docket Number: UTSDP3820WO- 1001380566injections. SC delivery resulted in much less detectable aptamer in plasma at all timepoints. By 120 min, and even more so by 240 min, there is very little aptamer that can still be detected by the qPCR assay.Example 6: In vivo effects on growth hormone (GH) secretion

[0169] Next, stimulation of ghrelin induced GH release was tested. Male C57BL / 6N mice, 12-14 weeks of age were purchased from Charles River for this study. The mice were single housed one week before the experiment. Further, mice were handled for three days prior to the study. Biotinylated anti-LEAP2 Aptamer 7 (1.4 pg / gram body weight or saline solution was administered i.p. twice, at 2 hours prior and again 30 minutes prior to the injection of ghrelin (Innovagen AB, Lund, Sweden; 1 mg / kg, s.c.). Ten minutes after the administration of the second injection of LEAP2 Aptamer 7 or saline, anesthesia with pentobarbital (50 mg / kg body weight, IP) was administered. Blood samples were obtained at t=5 min before and t=5 min (GH) and t=15 min after the administration of ghrelin or saline. The collected samples were immediately centrifuged at 4°C at 1,500 g for 15 minutes, and the plasma was collected and stored in a -80°C freezer. Plasma GH levels were determined using a GH ELISA kit(#EZRMGH-45K; Millipore-Merck). All data are presented as mean ± s.e.m. Two-way ANOVA followed by Tukey’s comparisons test were used for GH assay to test for significant differences among test groups.

[0170] As shown in FIG. 4A, animals treated with aptamer showed a greater stimulation (P=0.088) in ghrelin-induced GH release.Example 7: In vivo effects of aptamer administration on food intake

[0171] Two weeks after the growth hormone secretion studies, the mice were handled for three days to allow acclimatization. On the fourth day, food access was restricted for 2 hours before administering a subcutaneous injection of either saline or rat acyl-ghrelin. Next, biotinylated anti-LEAP2 Aptamer 7 (1.4 pg / gram body weight) or saline solution was administered intra-peritoneally twice, first injection 2 hours prior and then 30 minutes prior to the injection of ghrelin (Innovagen AB, Lund, Sweden; 0.5 mg / kg body weight, subcutaneously). Immediately after the ghrelin injection, food was reintroduced. A single preweighed standard chow pellet was placed on the cage floor, and the amount consumed during the following 1-hour and 2-hour periods was measured. As shown in FIGS. 5A and FIG. 5B, while ghrelin-induced food intake at 1 hour was not significantly increased over that induced by saline, treatment with LEAP2 aptamer led to a significant increase in ghrelin-induced food intake.301871278 - 61 -Atty. Docket Number: UTSDP3820WO- 1001380566Example 8: In vivo efficacy experiments

[0172] To test the in vivo efficacy of these novel reagents, 8-10 wk-old male C57BL / 6N (n=4 / group) were injected in the lateral tail vein with AAV with the LEAP2 shRNA construct 1 (4x1011GC / mL / Kg), PBS, or AAV with the scrambled shRNA. Injections were repeated 7d later. Five days later, qRT-PCR was performed in liver and jejunum, which have the greatest Leap2 expression and represent the major sources of circulating LEAP2. Leap2 mRNA was reduced by 82% in liver and 74% in jejunum (n=4 mice / group; p<0.01).Example 9: Summary

[0173] The anti-LEAP2 short hairpin (sh)RNA works by “knocking down” LEAP2 mRNA expression. The siDirect 2.0 software was used to design short hairpin RNA constructs that specifically silence Leap2 gene expression via RNA interference. These were then synthesized and a corresponding scrambled control version was generated as control. Stable transfection of Hepa1-6 mouse hepatoma cells with these constructs reduced Leap2 mRNA by 66.8% (construct 1) and 60.5% (construct 2) vs. cells transfected with scrambled control (n=3 experiments; p=0.07). The constructs were then packaged into an AAV serotype 8 viral delivery system (VectorBuilder) for in vivo Leap2 knockdown.

[0174] In vivo, biotinylated anti-LEAP2 aptamer 7 (1.4 pg / g body weight, IP) enhanced ghrelin-induced growth hormone release (P=0.088) and significantly increased ghrelin-induced food intake in male C57BL / 6N mice A validated qPCR-based detection assay confirmed comparable plasma aptamer levels following IV and IP administration, supporting IP as the preferred delivery route 3. Additionally, AAV-delivered LEAP2 shRNA reduced Leap2 mRNA by 82% in liver and 74% in jejunum (p<0.01), demonstrating the feasibility of a complementary gene silencing strategy301871278 - 62 -

Claims

Atty. Docket Number: UTSDP3820WO- 1001380566CLAIMSWhat is claimed is:

1. An anti-LEAP2 aptamer that specifically binds the liver expressed antimicrobial peptide 2 (LEAP2, LEAP-2) protein, or a derivative thereof.

2. The anti-LEAP2 aptamer of claim 1, wherein the aptamer comprises a nucleic acid sequence.

3. The anti-LEAP2 aptamer of claim 2, wherein the nucleic acid sequence is a deoxyribonucleic acid, or a variant thereof.

4. The anti-LEAP2 aptamer of claim 2, wherein the nucleic acid sequence comprises a sequence as set forth in SEQ ID NO: 2, or a sequence at least 95% identical thereto.

5. The anti-LEAP2 aptamer of claim 2, wherein the nucleic acid sequence comprises a sequence as set forth in SEQ ID NOs: 3-6, or a sequence at least 80% identical thereto.

6. The anti-LEAP2 aptamer of claim 2, wherein the nucleic acid sequence comprises a sequence as set forth in SEQ ID NOs: 7-10, or a sequence at least 80% identical thereto.

7. The anti-LEAP2 aptamer of any one of claims 1-6, wherein the aptamer further comprises one or more modifications.

8. The anti-LEAP2 aptamer of claim 7, wherein the one or more modifications comprise nucleobase modifications, sugar modifications, end modifications, or backbone modifications, or any combination thereof.

9. The anti-LEAP2 aptamer of claim 8, wherein the nucleobase modifications comprise incorporation of 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, di hydrouracil, beta-D-galactosylqueosine, inosine, N6-sopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methyl cytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid, 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, 2,6-301871278 - 63 -Atty. Docket Number: UTSDP3820WO- 1001380566diaminopurine, biotinylated analogs, 2'-Fluoro (2'-F) modification, 2'-O-Methyl (2'-OMe) modification, thiol groups, amine groups, or fluorescent labels, or any combination thereof.

10. The anti-LEAP2 aptamer of claim 8 or claim 9, wherein the sugar modifications comprise incorporation of 2'-Fluoro (2'-F), 2'-O-Methyl (2'-OMe), 2’-O-Methoxyethyl (2’-MOE), 2’-O-Amino (2’-NH2), arabinose, 2-fluoroarabinose, xylose, or hexose, or any combination thereof.

11. The anti-LEAP2 aptamer of any one of claims 8-10, wherein the one or more backbone modifications comprise incorporation of phosphorothioate, a phosphorodithioate, a phosphoramidothioate, a phosphoramidate, a phosphordiamidate, a methylphosphonate, an alkyl phosphotriester, formacetal PEGylation, circularization, spacer additions, multimerization, locked nucleic acids (LNA), unlocked nucleic acids, xeno nucleic acids, peptide nucleic acids (PNA), or end caps, or any combination thereof.

12. The anti-LEAP2 aptamer of claim 8, wherein the one or more modifications comprise 5’-biotinylation, 5’-PEGylation, 3’-PEGylation, or 5’-PEGylation and 3’-PEGylation.

13. A polynucleotide comprising the anti-LEAP2 aptamer of any one of claims 2-12.

14. A composition comprising one or more aptamers wherein the one or more aptamers specifically bind the liver expressed antimicrobial peptide 2 (LEAP2, LEAP-2) protein, or a derivative thereof.

15. The composition of claim 14, further comprising one or more anti-LEAP2 shRNAs.

16. The composition of claim 14 or claim 15, wherein the composition is a pharmaceutical composition, comprising at least one pharmaceutically acceptable excipient.

17. The composition of claim 16, wherein the administration of the pharmaceutical composition to a subject in need thereof reduces synthesis of LEAP2 protein, levels of LEAP2 protein, and / or LEAP2 binding with growth hormone secretagogue receptor (GHSR) in the subject.

18. The composition of claim 17, wherein the subject has or is suspected of having any one or more of cachexia (cancer cachexia, cardiac cachexia, pulmonary cachexia, renal cachexia, cachexia of aging), anorexia nervosa, hypoglycemia, depression, anxiety, Alzheimer’s disease, gastrointestinal dysmotility disorders (postoperative ileus, diabetic gastroparesis), and short stature.301871278 - 64 -Atty. Docket Number: UTSDP3820WO- 100138056619. The composition of any one of claims 16-18, wherein the pharmaceutical composition reduces ghrelin resistance and / or enhances the activity of endogenous ghrelin, or ghrelin-independent GHSR activity, and / or increases the activity of an administered ghrelin, and / or increases the activity of an administered GHSR agonist in the subject.

20. The composition of any one of claims 14-19, wherein the composition comprises a nanoparticle, a conjugate particle, a viral vector, a hydrogel, a liposome, an exosome, or a cell.

21. The composition of any one of claims 14-20, wherein the composition is for intravenous, intracranial, intrathecal, subcutaneous, intranasal, cranial, transmucosal, transnasal, transcranial, intracerebroventricular, intestinal, and / or parenteral delivery.

22. The composition of any one of claims 14-21, further comprising one or more GHSR agonists.

23. A method of treating a disease or a disorder in a subject in need thereof, the method comprising administering to the subject one or more anti-LEAP2 aptamers, and / or a composition thereof.

24. The method of claim 23, wherein the subject has or is suspected of having a ghrelin or growth hormone secretagogue receptor (GHSR) linked disease or disorder.

25. The method of claim 24, wherein the ghrelin or GHSR linked disease or disorder is a cachexia.

26. The method of claim 25, wherein the cachexia is a cancer cachexia, cardiac cachexia, pulmonary cachexia, renal cachexia, or cachexia of aging.

27. The method of claim 24, wherein the ghrelin or GHSR linked disease or disorder is anorexia nervosa, hypoglycemia, depression, anxiety, Alzheimer’s disease, dementia, age related memory decline, gastrointestinal dysmotility disorders (postoperative ileus, diabetic gastroparesis), hypoglycemia unawareness, post-traumatic stress disorder, amyotrophic lateral sclerosis or other neurodegenerative disorders, type 1 diabetes, type 2 diabetes, obesity, or short stature.

28. The method of any one of claims 23-27, wherein the method comprises administering to the subject the one or more anti-LEAP2 aptamers, and one or more anti-LEAP2 shRNAs.301871278 - 65 -Atty. Docket Number: UTSDP3820WO- 100138056629. The method of claim 28, wherein the one or more anti-LEAP2 aptamers, and the one or more anti-LEAP2 shRNAs are administered concurrently or separately.

30. The method of any one of claims 23-29, wherein the method further comprises administering one or more GHSR agonists, administering ghrelin or a functional variant thereof, increasing the activity of an administered ghrelin, or increasing the activity of an administered GHSR agonist, or any combination thereof.

31. The method of claim 30, wherein the GHSR agonist is administered concurrently or separately with the administration of the one or more anti-LEAP2 aptamers and / or the one or more anti-LEAP2 shRNAs.301871278 - 66 -

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