Compositions comprising synthetic mucins and uses thereof

Abstract

In one aspect, the disclosure relates to relates to pharmaceutical compositions that include copolymers and methods of treating gastrointestinal and other disorders associated with mucus depletion in a mucosal membrane. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

Description

ATTORNEY DOCKET NO.222112-2430 COMPOSITIONS COMPRISING SYNTHETIC MUCINS AND USES THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This Application claims the benefit of and priority to U.S. Provisional Application No. 63 / 653,036, filed on May 29, 2024 which is incorporated herein by reference in its entirety. BACKGROUND

[0002] The intestinal barrier includes the chemical barrier of the mucus layer, the mechanical barrier of the epithelial cell layer, and the immune barrier of the lamina propria. In a healthy individual, the intestinal epithelial barrier is highly selective to the luminal contents and does not permit the passage of microbial organisms or antigens. This physical barrier is enforced by a thick mucus layer lining the colonic epithelium and creating separation between microbes and enterocytes and paracellular tight junctions. This viscoelastic substance is structurally supported by heavily O-glycosylated mucin proteins, which act as a biological sieve and constitute only 1-5% of the aqueous gel. While the mucosal layer prevents microbial invasion, it forms a diffusion barrier in which small molecules, such as ions, water, nutrients, and gases, can readily diffuse through it and reach the epithelium.

[0003] A compromised mucus layer leaves mucosal membranes, especially the gut, at risk of a weakened defense against infections, increased inflammation, and damaged epithelial tissue. Increased intestinal permeability can result in the translocation of bacteria or their microbial products (e.g., lipopolysaccharides) to the bloodstream. Exploring mucin as a potential therapeutic target may aid immune tolerance, reduce bacterial translocation from the lumen into the intestinal epithelium, and consequently ameliorate gastrointestinal disorders. Historically, synthetic mucins have failed to replicate the intricate properties of natural mucins, and the therapeutic potential of synthetic mucins is effectively unexplored in vivo. Despite advances in research directed towards the design and synthesis of therapeutically useful and effective synthetic mucins, there remains a scarcity of compounds that can replicate the properties of natural mucins. These needs and other needs are satisfied by the present disclosure. SUMMARY

[0004] In accordance with the purpose(s) of the invention, as embodied and broadly described herein, the invention, in one aspect, relates to copolymer compounds that include mucoadhesive monomers, methods of making the same, pharmaceutical compositions comprising the same, and methods of treating gastrointestinal and other disorders associated with mucus depletion in a mucosal membrane.

[0005] Disclosed are pharmaceutical compositions comprising a therapeutically effective amount of at least one block copolymer, a pharmaceutically acceptable salt thereof, or aATTORNEY DOCKET NO.222112-2430 pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the block copolymer comprises a first block and a second block; where the first block can be selected from a mucoadhesive polymer block and a backbone polymer block; where the second block can be selected from a mucoadhesive polymer block and a backbone polymer block, provided that the first block and the second block are not the same; where the mucoadhesive polymer block can comprise at least one mucoadhesive monomer; where the backbone polymer block can comprise at least one backbone monomer; where the backbone polymer block does not contain a mucoadhesive monomer; where the mucoadhesive monomer can comprise at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and where the backbone monomer can comprise at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof.

[0006] Also disclosed are pharmaceutical compositions comprising a therapeutically effective amount of at least one random copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the random copolymer comprises: at least one mucoadhesive unit, where the mucoadhesive unit can comprise at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and at least one backbone unit, where the backbone unit can comprise at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof.

[0007] Also disclosed are pharmaceutical compositions comprising a therapeutically effective amount of at least one gradient copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the gradient copolymer comprises: at least one mucoadhesive unit, wherein the mucoadhesive unit can comprise at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and at least one backbone unit, wherein the backbone unit can comprise at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof; where greater than 50% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.ATTORNEY DOCKET NO.222112-2430

[0008] Also disclosed are pharmaceutical compositions comprising a therapeutically effective amount of one or more disclosed copolymer compounds, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0009] Also disclosed are methods for the treatment of a gastrointestinal disorder associated with mucus depletion in a mucosal membrane in a mammal comprising the step of administering to the mammal a therapeutically effective amount of at least one disclosed compound or pharmaceutically acceptable salt thereof.

[0010] Also disclosed are kits comprising at least one disclosed compound, or a pharmaceutically acceptable salt, hydrate, solvate, or polymorph thereof, and one or more of: (a) at least one agent known to treat a gastrointestinal condition; (b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane; (c) instructions for treating a gastrointestinal condition; (d) instructions for treating a disorder associated with mucus depletion in a mucosal membrane.

[0011] Also disclosed are uses of a disclosed compound, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a disorder associated with mucus depletion in a mucosal membrane in a mammal.

[0012] While aspects of the present disclosure can be described and claimed in a particular statutory class, such as the system statutory class, this is for convenience only and one of skill in the art will understand that each aspect of the present disclosure can be described and claimed in any statutory class. Unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification. BRIEF DESCRIPTION OF THE FIGURES

[0013] The accompanying figures, which are incorporated in and constitute a part of this specification, illustrate several aspects and, together with the description, serve to explain the principles of the disclosure.

[0014] FIG.1A shows a representative structure for copolymer AT1-104.

[0015] FIG.1B shows representative SEC-LS results for copolymer AT1-104 and calculated Mnand Đ.

[0016] FIG.1C shows a representative1H NMR spectrum for copolymer AT1-104.

[0017] FIG.2A shows a representative structure for copolymer AT1-105.ATTORNEY DOCKET NO.222112-2430

[0018] FIG.2B shows representative SEC-LS results for copolymer AT1-105 and calculated Mnand Đ.

[0019] FIG.2C shows a representative1H NMR spectrum for copolymer AT1-105.

[0020] FIG.3A shows representative SEC-LS results for copolymer AT1-106[macroCTA] and calculated Mnand Đ.

[0021] FIG.3B shows representative SEC-LS results for copolymer AT1-106 and calculated Mnand Đ.

[0022] FIG.3C shows a representative1H NMR spectrum for copolymer AT1-106.

[0023] FIG.4A shows representative SEC-LS results for copolymer AT1-111[macroCTA] and calculated Mnand Đ.

[0024] FIG.4B shows a representative1H NMR spectrum for copolymer AT1-111.

[0025] FIG.5 shows representative SEC-LS results for copolymer JA1-032 and calculated Mnand Đ.

[0026] FIG.6A shows representative SEC-LS results for copolymer AT1-114 and calculated Mnand Đ

[0027] FIG.6B shows a representative1H NMR spectrum for copolymer AT1-114.

[0028] FIG.7 shows a representative1H NMR spectrum for copolymer AT1-118.

[0029] FIGS.8A-8E show a representative Shear-Labile Interaction Polymers (SLIPs) design strategy: photoiniferter polymerization (FIG.8A) leads to ultra-high molecular weight, water- soluble ABA triblock copolymers, where (FIG.8B) terminal A blocks contain functionality for reversible crosslinking, and the central B block is composed of units that promote hydration. This approach leads to well-defined polymers with targeted molecular weights up to 10 MDa (FIG.8C). The resulting SLIPs (orange) have sizes on the order of natural mucins (blue) (FIG. 8D) and are expected to form gels with comparable mesh sizes. The polymerization approach is capable of yielding polymers with controlled architecture (FIG.8E) for fine tuning rheological properties and gel behavior.

[0030] FIG.9A shows representative viability data for hTCEpi cells after 5-day exposure to SLIP backbone polymer (PDMA) with two molecular sizes (1.5 and 5 MDa) and at three concentrations (1.8, 18 and 90 mg / mL). PDMA-EYA = backbone with fluorescent label attached to allow tracking.

[0031] FIG.9B shows a representative 3-Dimentional confocal image of epithelial (hTCEpi) cells protected from E. coli by a layer of SLIPs. Glass bottom imaging dishes were seeded at 0.5 million cells per mL. Cells were stained with CellTracker™ Deep Red (Invitrogen™ C34565). The dish was cultured for 2 days to reach confluence before the experiments. E. coli was separately cultured and stained with BactoView™ Live Red (Biotium™, 40101-T). The fully confluent dishes were incubated for 1 h with a solution of SLIPs 1.5 %wt. labelled with Fluorescein acrylamide, prior to E. coli addition.ATTORNEY DOCKET NO.222112-2430

[0032] FIGS. 10A-10B show representative body weight change data (FIG. 10A) and a representative composite image of the small intestine of Wild-type C57BL / 6J mice (FIG.10B) treated with SLIPs for 10 days. Tissue was stained with MUC2 Monoclonal Antibody (HL1724) (red-GFP 470 / 525 nm), Nuclei were stained with DAPI (blue-357 / 447 nm) and SLIPs were labeled with fluorescein (green-531 / 593 nm). Image was visualized and stitched with the EVOS M7000 Imaging System. Magnification: 20X.

[0033] FIGS. 10C-10F show bacterial diversity from stool samples (FIG. 10C); cumulative excretion of SLIPs throughout the feces (FIG. 10D); relative abundance of the 4 most abundant phylum (FIG.10E); and relative abundance of the 10 most abundant species of Wild-type C57BL / 6J mice (FIG. 10F). SLIPs were added to drinking water in 6 different concentrations for 10 days. Stool samples were obtained every other day. Microbial DNA was extracted, and the full length of the 16S rRNA was amplified and sequenced. Shannon diversity was calculated after rarefying the dataset to the same number of sequences. After verifying that the data was not normally distributed, differences were calculated by the non- parametric Wilcox test. Differential abundance among treatments was calculated by Aldex2 (ANOVA-Like Differential Expression tool for compositional data). Data are an average of three biological repetitions per treatment, n = 18.

[0034] FIGS.11A-11I show representative efficacy data for use of a disclosed SLIPs in Agr2- / -mice lacking mucus production through showing body weight changes over time (FIG.11A); representative changes in body composition (FIG. 11B) performed DEXA (dual x-ray absorptiometry) scans; fluorescein isothiocyanate conjugated (FITC)-Dextran assay (FIG. 11C) showing representative reduction of gut permeability; changes in bacterial diversity from stool samples (FIG. 11D); abundance of Enterobacteriaceae in stool samples (FIG. 11E); showing representative immune response data (FIG. 11F) showing representative data comprising quantitation by measuring the plasma samples with the panel ProcartaPlex™ Mouse Th1 / Th2 Cytokine, and the ProcartaPlex Mouse Cytokine Panel 1B with the Luminex xMAP technology; histological changes (FIG.11G) and macroscopic appearances (FIG.11H) of colon tissues H&E stained in Swiss roll; and representative images of DEXA scans (FIG. 11I). SLIPs were added to the drinking water of the Agr2- / -and Wild-type mice (8 weeks old) at zero (control) and 40 mg of SLIPs (treatment) for 10 days. Weight was measured every other day during the intervention and fecal samples were obtained every other day from the start to end of the experiments. Data are averages of six biological repetitions per treatment.

[0035] FIGS. 12A-12D show representative data as follows: a proteome profile of spleen (FIGS.12A and 12B) and colon (FIGS.12C and 12D) samples from Agr2- / -mice treated and untreated with SLIPs: PCA plot showing a clear clustering of samples according to the experimental groups (FIGS. 12A and 12C) and volcano plot showing the differentially expressed proteins between treatments (FIGS.12B and 12D). Significant differences wereATTORNEY DOCKET NO.222112-2430 defined using FDR-adjusted p-values <= 0.01 and Fold Change >= 2. N=4 samples per treatment

[0036] FIG. 13 shows a representative heatmap with the main differentially expressed proteins from spleen samples of Agr2- / -mice involved in the inflammatory response altered by the treatment. Significant differences are defined using FDR-adjusted p-values of 0.01 and Fold Change >= 2. N=4 samples per treatment.

[0037] FIG.14 shows a representative boxplot from colon samples of Agr2- / -mice illustrating the abundance of proteins involved in the immunological response to lipopolysaccharide (LPS). The dataset was normalized by the median and transformed using log10 transformation and range scaling (mean-centered and divided by the range of each variable). Significant differences were defined using FDR-adjusted p-values <= 0.01 and Fold Change >= 2. N=4 samples per treatment.

[0038] FIGS. 15A and 15B show microbial degradability of representative SLIPs via an indirect growth measurement in liquid media with porcine mucin (FIG.15A) or SLIPs (FIG. 15B) as a single carbon source inoculated with fresh mice fecal pellets. Means with the same letter are not significantly different (p-value 0.05, ANOVA followed by Tukey test).

[0039] FIG.16 shows a detection of representative fluorescein-labeled SLIPs over time during treatment (up to 144 hours) and after treatment.

[0040] FIGS.17A-17B shows glucose diffusion across SLIPs at various pH conditions. Data points represent the mean percentage of glucose diffused at each pH, with error bars indicating standard deviations. Means with the same letter are not significantly different (p- value 0.05, ANOVA followed by Tukey test).

[0041] FIG.18 shows a schematic of glucose diffusion across the gastrointestinal tract. This illustrates glucose diffusion percentages across different gastrointestinal (GI) tract regions, correlated with the pH conditions tested and the physiological pH levels observed in each region. The diffusion ratios were converted into percentage values, and the average percentage per pH condition was mapped to their respective GI locations. pH conditions of the GI tract were determined based on available literature.

[0042] FIG.19A shows a schematic illustration of a representative fluorometric approach for detecting unbound FITC-LPS.

[0043] FIG.19B shows a representative standard curve prepared with known concentrations of FITC-LPS.

[0044] FIG. 20 shows a representative dose-response relationship of SLIPs binding to bacterial LPS. The mean binding capacity of SLIPs (8.3 mg / mL) was measured across LPS concentrations. Experimental data points (black circles) represent mean values ± standard error (error bars). A logistic regression model (blue line) was fitted to the data.ATTORNEY DOCKET NO.222112-2430

[0045] FIG. 21 shows differentially abundant species between representative SLIP-treated KO mice and KO controls over the course of 17 days.

[0046] FIGS. 22A and 22B show relative abundance changes in anaerobic species (FIG. 22A) and facultative anaerobic species (FIG. 22B) found in the fecal microbiome due to representative SLIP treatment in KO and WT mice.

[0047] FIGS.23A-23C show a representative effect of SLIPs on the colon proteome of Agr2- / - mice (KO for mucus production) treated with SLIPs, untreated controls, and wild-type mice over 17 days, illustrating significantly negatively enriched Gene Ontology (GO) Biological Functions (FIG.23A); a heat map of selected pro- and anti-inflammatory upstream cytokines inferred as z-scores, showing up-regulation (red) or down-regulation (blue) (*P < 0.05, Z = ±1.96; **P < 0.01; ***P < 0.001) (FIG. 23B); and a modeled mechanism of inflammatory regulation driven by SLIPs (FIG.23C) where proteins are overlaid with z-scores, indicating inhibition (blue) or activation (red) and dashed lines represent indirect relationships, while solid lines indicate direct relationships.

[0048] FIG. 24 shows representative body weight change throughout treatment. Mice (3 females and 3 males per group) were treated with DSS for 4 days, and the treatment group received 6.6 mg / mL of SLIPs in the drinking water, while the control group received only water. Error bars represent the standard error of the mean. A Wilcoxon rank-sum test indicated a significant difference in weight at day 8 (p < 0.05), and the corresponding effect size (r = 0.6) suggested a large effect.

[0049] FIG.25 shows a representative disease activity index throughout treatment. Mice (3 females and 3 males per group) were treated with DSS for 4 days, and the treatment group received 6.6 mg / mL of SLIPs in the drinking water, while the control group received only water. Error bars represent the standard error of the mean. A Wilcoxon rank-sum test indicated a significant difference in the disease activity index at day 8 (p < 0.05), and the corresponding effect size (r = 0.5) suggested a large effect.

[0050] FIG.26 shows a representative gut permeability assessment measured by the FITC- dextran 4 kDa in serum from mice after SLIPs treatment. A Wilcoxon rank-sum test indicated a significant difference in permeability at day 8 (p < 0.05), and the corresponding effect size (r = 0.66) suggested a large effect.

[0051] Additional advantages of the disclosure will be set forth in part in the description which follows, and in part will be obvious from the description, or can be learned by practice of the disclosure. The advantages of the disclosure will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the disclosure, as claimed.ATTORNEY DOCKET NO.222112-2430 DETAILED DESCRIPTION

[0052] Many modifications and other embodiments disclosed herein will come to mind to one skilled in the art to which the disclosed compositions and methods pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the disclosures are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. The skilled artisan will recognize many variants and adaptations of the aspects described herein. These variants and adaptations are intended to be included in the teachings of this disclosure and to be encompassed by the claims herein.

[0053] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

[0054] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure.

[0055] Any recited method can be carried out in the order of events recited or in any other order that is logically possible. That is, unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

[0056] All publications and patents cited in this specification are cited to disclose and describe the methods and / or materials in connection with which the publications are cited. All such publications and patents are herein incorporated by references as if each individual publication or patent were specifically and individually indicated to be incorporated by reference. Such incorporation by reference is expressly limited to the methods and / or materials described in the cited publications and patents and does not extend to any lexicographical definitions from the cited publications and patents. Any lexicographical definition in the publications and patents cited that is not also expressly repeated in the instant application should not be treated as such and should not be read as defining any terms appearing in the accompanying claims. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided could beATTORNEY DOCKET NO.222112-2430 different from the actual publication dates that may need to be independently confirmed.

[0057] While aspects of the present disclosure can be described and claimed in a particular statutory class, such as the system statutory class, this is for convenience only and one of skill in the art will understand that each aspect of the present disclosure can be described and claimed in any statutory class.

[0058] It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed compositions and methods belong. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly defined herein.

[0059] Aspects of the present disclosure will employ, unless otherwise indicated, techniques of molecular biology, microbiology, organic chemistry, biochemistry, physiology, cell biology, blood vessel biology, and the like, which are within the skill of the art. Such techniques are explained fully in the literature.

[0060] Prior to describing the various aspects of the present disclosure, the following definitions are provided and should be used unless otherwise indicated. Additional terms may be defined elsewhere in the present disclosure. A. Definitions

[0061] As used herein, “comprising” is to be interpreted as specifying the presence of the stated features, integers, steps, or components as referred to, but does not preclude the presence or addition of one or more features, integers, steps, or components, or groups thereof. Moreover, each of the terms “by”, “comprising,” “comprises”, “comprised of,” “including,” “includes,” “included,” “involving,” “involves,” “involved,” and “such as” are used in their open, non-limiting sense and may be used interchangeably. Further, the term “comprising” is intended to include examples and aspects encompassed by the terms “consisting essentially of” and “consisting of.” Similarly, the term “consisting essentially of” is intended to include examples encompassed by the term “consisting of.

[0062] As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items. Expressions such as “at least one of,” when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.

[0063] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a polymer block,” “a monomer,” or “a residue,” include, but are not limited to, twoATTORNEY DOCKET NO.222112-2430 or more such polymer blocks, monomers, or residues, including combinations of polymer blocks, monomers, or residues, and the like.

[0064] It should be noted that ratios, concentrations, amounts, and other numerical data can be expressed herein in a range format. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms a further aspect. For example, if the value “about 10” is disclosed, then “10” is also disclosed.

[0065] Where a range is expressed, a further aspect includes from the one particular value and / or to the other particular value. Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure. For example, where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure, e.g. the phrase “x to y” includes the range from ‘x’ to ‘y’ as well as the range greater than ‘x’ and less than ‘y’. The range can also be expressed as an upper limit, e.g. ‘about x, y, z, or less’ and should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘less than x’, less than y’, and ‘less than z’. Likewise, the phrase ‘about x, y, z, or greater’ should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘greater than x’, greater than y’, and ‘greater than z’. In addition, the phrase “about ‘x’ to ‘y’”, where ‘x’ and ‘y’ are numerical values, includes “about ‘x’ to about ‘y’”.

[0066] It is to be understood that such a range format is used for convenience and brevity, and thus, should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. To illustrate, a numerical range of “about 0.1% to 5%” should be interpreted to include not only the explicitly recited values of about 0.1% to about 5%, but also include individual values (e.g., about 1%, about 2%, about 3%, and about 4%) and the sub-rangesATTORNEY DOCKET NO.222112-2430 (e.g., about 0.5% to about 1.1%; about 5% to about 2.4%; about 0.5% to about 3.2%, and about 0.5% to about 4.4%, and other possible sub-ranges) within the indicated range.

[0067] As used herein, "about," "approximately," “substantially,” and the like, when used in connection with a numerical variable, can generally refers to the value of the variable and to all values of the variable that are within the experimental error (e.g., within the 95% confidence interval for the mean) or within + / - 10% of the indicated value, whichever is greater. As used herein, the terms “about,” “approximate,” “at or about,” and “substantially” can mean that the amount or value in question can be the exact value or a value that provides equivalent results or effects as recited in the claims or taught herein. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact, but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art such that equivalent results or effects are obtained. In some circumstances, the value that provides equivalent results or effects cannot be reasonably determined. In general, an amount, size, formulation, parameter or other quantity or characteristic is “about,” “approximate,” or “at or about” whether or not expressly stated to be such. It is understood that where “about,” “approximate,” or “at or about” is used before a quantitative value, the parameter also includes the specific quantitative value itself, unless specifically stated otherwise.

[0068] As used herein, the terms “optional” or “optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0069] As used herein, “administering” can refer to an administration that is oral, topical, intravenous, subcutaneous, transcutaneous, transdermal, intramuscular, intra-joint, intra- arteriole, intradermal, intraventricular, intraosseous, intraocular, intracranial, intraperitoneal, intralesional, intranasal, intracardiac, intraarticular, intracavernous, intrathecal, intraviral, intracerebral, and intracerebroventricular, intratympanic, intracochlear, rectal, vaginal, by inhalation, by catheters, stents or via an implanted reservoir or other device that administers, either actively or passively (e.g. by diffusion) a composition the perivascular space and adventitia. For example a medical device such as a stent can contain a composition or formulation disposed on its surface, which can then dissolve or be otherwise distributed to the surrounding tissue and cells. Administration can be continuous or intermittent. In various aspects, a preparation can be administered therapeutically; that is, administered to treat an existing disease or condition. In further various aspects, a preparation can be administered prophylactically; that is, administered for prevention of a disease or condition.

[0070] As used herein, “therapeutic agent” can refer to any substance, compound, molecule, and the like, which can be biologically active or otherwise can induce a pharmacologic,ATTORNEY DOCKET NO.222112-2430 immunogenic, biologic and / or physiologic effect on a subject to which it is administered to by local and / or systemic action. A therapeutic agent can be a primary active agent, or in other words, the component(s) of a composition to which the whole or part of the effect of the composition is attributed. A therapeutic agent can be a secondary therapeutic agent, or in other words, the component(s) of a composition to which an additional part and / or other effect of the composition is attributed. The term therefore encompasses those compounds or chemicals traditionally regarded as drugs, vaccines, and biopharmaceuticals including molecules such as proteins, peptides, hormones, nucleic acids, gene constructs and the like. Examples of therapeutic agents are described in well-known literature references such as the Merck Index (14th edition), the Physicians' Desk Reference (64th edition), and The Pharmacological Basis of Therapeutics (12th edition), and they include, without limitation, medicaments; vitamins; mineral supplements; substances used for the treatment, prevention, diagnosis, cure or mitigation of a disease or illness; substances that affect the structure or function of the body, or pro-drugs, which become biologically active or more active after they have been placed in a physiological environment. For example, the term “therapeutic agent” includes compounds or compositions for use in all of the major therapeutic areas including, but not limited to, adjuvants; anti-infectives such as antibiotics and antiviral agents; analgesics and analgesic combinations, anorexics, anti-inflammatory agents, anti-epileptics, local and general anesthetics, hypnotics, sedatives, antipsychotic agents, neuroleptic agents, antidepressants, anxiolytics, antagonists, neuron blocking agents, anticholinergic and cholinomimetic agents, antimuscarinic and muscarinic agents, antiadrenergics, antiarrhythmics, antihypertensive agents, hormones, and nutrients, antiarthritics, antiasthmatic agents, anticonvulsants, antihistamines, antinauseants, antineoplastics, antipruritics, antipyretics; antispasmodics, cardiovascular preparations (including calcium channel blockers, beta-blockers, beta-agonists and antiarrhythmics), antihypertensives, diuretics, vasodilators; central nervous system stimulants; cough and cold preparations; decongestants; diagnostics; hormones; bone growth stimulants and bone resorption inhibitors; immunosuppressives; muscle relaxants; psychostimulants; sedatives; tranquilizers; proteins, peptides, and fragments thereof (whether naturally occurring, chemically synthesized or recombinantly produced); and nucleic acid molecules (polymeric forms of two or more nucleotides, either ribonucleotides (RNA) or deoxyribonucleotides (DNA) including both double- and single-stranded molecules, gene constructs, expression vectors, antisense molecules and the like), small molecules (e.g., doxorubicin) and other biologically active macromolecules such as, for example, proteins and enzymes. The agent may be a biologically active agent used in medical, including veterinary, applications and in agriculture, such as with plants, as well as other areas. The term therapeutic agent also includes without limitation, medicaments; vitamins; mineral supplements; substances used for the treatment, prevention,ATTORNEY DOCKET NO.222112-2430 diagnosis, cure or mitigation of disease or illness; or substances which affect the structure or function of the body; or pro- drugs, which become biologically active or more active after they have been placed in a predetermined physiological environment.

[0071] As used herein, “kit” means a collection of at least two components constituting the kit. Together, the components constitute a functional unit for a given purpose. Individual member components may be physically packaged together or separately. For example, a kit comprising an instruction for using the kit may or may not physically include the instruction with other individual member components. Instead, the instruction can be supplied as a separate member component, either in a paper form or an electronic form which may be supplied on computer readable memory device or downloaded from an internet website, or as recorded presentation.

[0072] As used herein, “instruction(s)” means documents describing relevant materials or methodologies pertaining to a kit. These materials may include any combination of the following: background information, list of components and their availability information (purchase information, etc.), brief or detailed protocols for using the kit, trouble-shooting, references, technical support, and any other related documents. Instructions can be supplied with the kit or as a separate member component, either as a paper form or an electronic form which may be supplied on computer readable memory device or downloaded from an internet website, or as recorded presentation. Instructions can comprise one or multiple documents, and are meant to include future updates.

[0073] As used herein, “attached” can refer to covalent or non-covalent interaction between two or more molecules. Non-covalent interactions can include ionic bonds, electrostatic interactions, van der Walls forces, dipole-dipole interactions, dipole-induced-dipole interactions, London dispersion forces, hydrogen bonding, halogen bonding, electromagnetic interactions, π-π interactions, cation-π interactions, anion-π interactions, polar π-interactions, and hydrophobic effects.

[0074] As used herein, the term “subject” can be a vertebrate, such as a mammal, a fish, a bird, a reptile, or an amphibian. Thus, the subject of the herein disclosed methods can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The term does not denote a particular age or sex. Thus, adult and juvenile subjects, whether male or female, are intended to be covered. In one aspect, the subject is a mammal. A patient refers to a subject afflicted with a disease or disorder. The term “patient” includes human and veterinary subjects.

[0075] As used herein, the terms "treating" and "treatment" can refer generally to obtaining a desired pharmacological and / or physiological effect. The effect can be, but does not necessarily have to be, prophylactic in terms of preventing or partially preventing a disease, symptom or condition thereof, such as a gastrointestinal disorder, a disorder or diseaseATTORNEY DOCKET NO.222112-2430 associated with mucus depletion in a mucosal membrane, cancer, and / or an enteric infection. The effect can be therapeutic in terms of a partial or complete cure of a disease, condition, symptom or adverse effect attributed to the disease, disorder, or condition. The term "treatment" as used herein can include any treatment of a gastrointestinal disorder, a disorder or disease associated with mucus depletion in a mucosal membrane, cancer, and / or an enteric infection in a subject, particularly a human and can include any one or more of the following: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., mitigating or ameliorating the disease and / or its symptoms or conditions. The term "treatment" as used herein can refer to both therapeutic treatment alone, prophylactic treatment alone, or both therapeutic and prophylactic treatment. Those in need of treatment (subjects in need thereof) can include those already with the disorder and / or those in which the disorder is to be prevented. As used herein, the term "treating", can include inhibiting the disease, disorder or condition, e.g., impeding its progress; and relieving the disease, disorder, or condition, e.g., causing regression of the disease, disorder and / or condition. Treating the disease, disorder, or condition can include ameliorating at least one symptom of the particular disease, disorder, or condition, even if the underlying pathophysiology is not affected, e.g., such as treating the pain of a subject by administration of an analgesic agent even though such agent does not treat the cause of the pain.

[0076] As used herein, “dose,” “unit dose,” or “dosage” can refer to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of a disclosed compound and / or a pharmaceutical composition thereof calculated to produce the desired response or responses in association with its administration.

[0077] As used herein, “therapeutic” can refer to treating, healing, and / or ameliorating a disease, disorder, condition, or side effect, or to decreasing in the rate of advancement of a disease, disorder, condition, or side effect.

[0078] As used herein, “effective amount” can refer to the amount of a disclosed compound or pharmaceutical composition provided herein that is sufficient to effect beneficial or desired biological, emotional, medical, or clinical response of a cell, tissue, system, animal, or human. An effective amount can be administered in one or more administrations, applications, or dosages. The term can also include within its scope amounts effective to enhance or restore to substantially normal physiological function.

[0079] As used herein, the term “therapeutically effective amount” refers to an amount that is sufficient to achieve the desired therapeutic result or to have an effect on undesired symptoms, but is generally insufficient to cause adverse side effects. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorderATTORNEY DOCKET NO.222112-2430 being treated and the severity of the disorder; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration; the route of administration; the rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed and like factors within the knowledge and expertise of the health practitioner and which may be well known in the medical arts. In the case of treating a particular disease or condition, in some instances, the desired response can be inhibiting the progression of the disease or condition. This may involve only slowing the progression of the disease temporarily. However, in other instances, it may be desirable to halt the progression of the disease permanently. This can be monitored by routine diagnostic methods known to one of ordinary skill in the art for any particular disease. The desired response to treatment of the disease or condition also can be delaying the onset or even preventing the onset of the disease or condition.

[0080] For example, it is well within the skill of the art to start doses of a compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. If desired, the effective daily dose can be divided into multiple doses for purposes of administration. Consequently, single dose compositions can contain such amounts or submultiples thereof to make up the daily dose. The dosage can be adjusted by the individual physician in the event of any contraindications. It is generally preferred that a maximum dose of the pharmacological agents of the invention (alone or in combination with other therapeutic agents) be used, that is, the highest safe dose according to sound medical judgment. It will be understood by those of ordinary skill in the art however, that a patient may insist upon a lower dose or tolerable dose for medical reasons, psychological reasons or for virtually any other reasons.

[0081] A response to a therapeutically effective dose of a disclosed compound and / or pharmaceutical composition, for example, can be measured by determining the physiological effects of the treatment or medication, such as the decrease or lack of disease symptoms following administration of the treatment or pharmacological agent. Other assays will be known to one of ordinary skill in the art and can be employed for measuring the level of the response. The amount of a treatment may be varied for example by increasing or decreasing the amount of a disclosed compound and / or pharmaceutical composition, by changing the disclosed compound and / or pharmaceutical composition administered, by changing the route of administration, by changing the dosage timing and so on. Dosage can vary, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of pharmaceutical products.

[0082] As used herein, the term “prophylactically effective amount” refers to an amount effective for preventing onset or initiation of a disease or condition.ATTORNEY DOCKET NO.222112-2430

[0083] As used herein, the term “prevent” or “preventing” refers to precluding, averting, obviating, forestalling, stopping, or hindering something from happening, especially by advance action. It is understood that where reduce, inhibit or prevent are used herein, unless specifically indicated otherwise, the use of the other two words is also expressly disclosed.

[0084] The term “pharmaceutically acceptable” describes a material that is not biologically or otherwise undesirable, i.e., without causing an unacceptable level of undesirable biological effects or interacting in a deleterious manner.

[0085] The term “pharmaceutically acceptable salts”, as used herein, means salts of the active principal agents which are prepared with acids or bases that are tolerated by a biological system or tolerated by a subject or tolerated by a biological system and tolerated by a subject when administered in a therapeutically effective amount. When compounds of the present disclosure contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include, but are not limited to; sodium, potassium, calcium, ammonium, organic amino, magnesium salt, lithium salt, strontium salt or a similar salt. When compounds of the present disclosure contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include, but are not limited to; those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like.

[0086] The term “pharmaceutically acceptable ester” refers to esters of compounds of the present disclosure which hydrolyze in vivo and include those that break down readily in the human body to leave the parent compound or a salt thereof. Examples of pharmaceutically acceptable, non-toxic esters of the present disclosure include C 1 -to-C 6 alkyl esters and C 5 -to-C 7 cycloalkyl esters, although C 1 -to-C 4 alkyl esters are preferred. Esters of disclosed compounds can be prepared according to conventional methods. Pharmaceutically acceptable esters can be appended onto hydroxy groups by reaction of the compound that contains the hydroxy group with acid and an alkylcarboxylic acid such as acetic acid, or with acid and an arylcarboxylic acid such as benzoic acid. In the case of compounds containing carboxylic acid groups, the pharmaceutically acceptable esters are prepared from compoundsATTORNEY DOCKET NO.222112-2430 containing the carboxylic acid groups by reaction of the compound with base such as triethylamine and an alkyl halide, for example with methyl iodide, benzyl iodide, cyclopentyl iodide or alkyl triflate. They also can be prepared by reaction of the compound with an acid such as hydrochloric acid and an alcohol such as ethanol or methanol.

[0087] The term “pharmaceutically acceptable amide” refers to non-toxic amides of the present disclosure derived from ammonia, primary C 1 -to-C 6 alkyl amines and secondary C 1 -to-C 6 dialkyl amines. In the case of secondary amines, the amine can also be in the form of a 5- or 6-membered heterocycle containing one nitrogen atom. Amides derived from ammonia, C 1 -to-C 3 alkyl primary amides and C 1 -to-C 2 dialkyl secondary amides are preferred. Amides of disclosed compounds can be prepared according to conventional methods. Pharmaceutically acceptable amides can be prepared from compounds containing primary or secondary amine groups by reaction of the compound that contains the amino group with an alkyl anhydride, aryl anhydride, acyl halide, or aroyl halide. In the case of compounds containing carboxylic acid groups, the pharmaceutically acceptable amides are prepared from compounds containing the carboxylic acid groups by reaction of the compound with base such as triethylamine, a dehydrating agent such as dicyclohexyl carbodiimide or carbonyl diimidazole, and an alkyl amine, dialkylamine, for example with methylamine, diethylamine, and piperidine. They also can be prepared by reaction of the compound with an acid such as sulfuric acid and an alkylcarboxylic acid such as acetic acid, or with acid and an arylcarboxylic acid such as benzoic acid under dehydrating conditions such as with molecular sieves added. The composition can contain a compound of the present disclosure in the form of a pharmaceutically acceptable prodrug.

[0088] The term “pharmaceutically acceptable prodrug” or “prodrug” represents those prodrugs of the compounds of the present disclosure which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for their intended use. Prodrugs of the present disclosure can be rapidly transformed in vivo to a parent compound having a structure of a disclosed compound, for example, by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, V. 14 of the A.C.S. Symposium Series, and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987).

[0089] As used herein, the term “derivative” refers to a compound having a structure derived from the structure of a parent compound (e.g., a compound disclosed herein) and whose structure is sufficiently similar to those disclosed herein and based upon that similarity, would be expected by one skilled in the art to exhibit the same or similar activities and utilities as the claimed compounds, or to induce, as a precursor, the same or similar activities and utilities asATTORNEY DOCKET NO.222112-2430 the claimed compounds. Exemplary derivatives include salts, esters, amides, salts of esters or amides, and N-oxides of a parent compound.

[0090] As used herein, nomenclature for compounds, including organic compounds, can be given using common names, IUPAC, IUBMB, or CAS recommendations for nomenclature. When one or more stereochemical features are present, Cahn-Ingold-Prelog rules for stereochemistry can be employed to designate stereochemical priority, E / Z specification, and the like. One of skill in the art can readily ascertain the structure of a compound if given a name, either by systemic reduction of the compound structure using naming conventions, or by commercially available software, such as CHEMDRAW™ (Cambridgesoft Corporation, U.S.A.).

[0091] The term “alkyl” as used herein is a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s- butyl, t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like. The alkyl group can be cyclic or acyclic. The alkyl group can be branched or unbranched. The alkyl group can also be substituted or unsubstituted. For example, the alkyl group can be substituted with one or more groups including, but not limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol, as described herein. A “lower alkyl” group is an alkyl group containing from one to six (e.g., from one to four) carbon atoms. The term alkyl group can also be a C1 alkyl, C1-C2 alkyl, C1-C3 alkyl, C1-C4 alkyl, C1-C5 alkyl, C1-C6 alkyl, C1-C7 alkyl, C1-C8 alkyl, C1-C9 alkyl, C1-C10 alkyl, and the like up to and including a C1-C24 alkyl.

[0092] Throughout the specification “alkyl” is generally used to refer to both unsubstituted alkyl groups and substituted alkyl groups; however, substituted alkyl groups are also specifically referred to herein by identifying the specific substituent(s) on the alkyl group. For example, the term “halogenated alkyl” or “haloalkyl” specifically refers to an alkyl group that is substituted with one or more halide, e.g., fluorine, chlorine, bromine, or iodine. Alternatively, the term “monohaloalkyl” specifically refers to an alkyl group that is substituted with a single halide, e.g. fluorine, chlorine, bromine, or iodine. The term “polyhaloalkyl” specifically refers to an alkyl group that is independently substituted with two or more halides, i.e. each halide substituent need not be the same halide as another halide substituent, nor do the multiple instances of a halide substituent need to be on the same carbon. The term “alkoxyalkyl” specifically refers to an alkyl group that is substituted with one or more alkoxy groups, as described below. The term “aminoalkyl” specifically refers to an alkyl group that is substituted with one or more amino groups. The term “hydroxyalkyl” specifically refers to an alkyl group that is substituted with one or more hydroxy groups. When “alkyl” is used in one instance and a specific term such as “hydroxyalkyl” is used in another, it is not meant to imply that the term “alkyl” does not also refer to specific terms such as “hydroxyalkyl” and the like.ATTORNEY DOCKET NO.222112-2430

[0093] This practice is also used for other groups described herein. That is, while a term such as “cycloalkyl” refers to both unsubstituted and substituted cycloalkyl moieties, the substituted moieties can, in addition, be specifically identified herein; for example, a particular substituted cycloalkyl can be referred to as, e.g., an “alkylcycloalkyl.” Similarly, a substituted alkoxy can be specifically referred to as, e.g., a “halogenated alkoxy,” a particular substituted alkenyl can be, e.g., an “alkenylalcohol,” and the like. Again, the practice of using a general term, such as “cycloalkyl,” and a specific term, such as “alkylcycloalkyl,” is not meant to imply that the general term does not also include the specific term.

[0094] The term “thiol” as used herein is represented by the formula —SH.

[0095] As described herein, compounds of the disclosure may contain “optionally substituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds. In is also contemplated that, in certain aspects, unless expressly indicated to the contrary, individual substituents can be further optionally substituted (i.e., further substituted or unsubstituted).

[0096] A residue of a chemical species, as used in the specification and concluding claims, refers to the moiety that is the resulting product of the chemical species in a particular reaction scheme or subsequent formulation or chemical product, regardless of whether the moiety is actually obtained from the chemical species. Thus, a carboxylic acid residue in a compound refers to one or more −C(O)OH units in the compound, regardless of whether a carboxylic acid was used to prepare the compound. Similarly, a phenylboronic acid residue in a compound refers to one or more −PhB(OH)2moieties in the compound, regardless of whether the residue is obtained by reacting phenylboronic acid or a derivative thereof to obtain the compound.

[0097] A very close synonym of the term “residue” is the term “radical,” which as used in the specification and concluding claims, refers to a fragment, group, or substructure of a molecule described herein, regardless of how the molecule is prepared. For example, a 2,4- thiazolidinedione radical in a particular compound has the structure:ATTORNEY DOCKET NO.222112-2430

[0098] regardless of whether thiazolidinedione is used to prepare the compound. In some embodiments the radical (for example an alkyl) can be further modified (i.e., substituted alkyl) by having bonded thereto one or more “substituent radicals.” The number of atoms in a given radical is not critical to the present disclosure unless it is indicated to the contrary elsewhere herein.

[0099] Compounds described herein can contain one or more double bonds and, thus, potentially give rise to cis / trans (E / Z) isomers, as well as other conformational isomers. Unless stated to the contrary, the disclosure includes all such possible isomers, as well as mixtures of such isomers.

[0100] Unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, e.g., each enantiomer and diastereomer, and a mixture of isomers, such as a racemic or scalemic mixture. Compounds described herein can contain one or more asymmetric centers and, thus, potentially give rise to diastereomers and optical isomers. Unless stated to the contrary, the present disclosure includes all such possible diastereomers as well as their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers, and pharmaceutically acceptable salts thereof. Mixtures of stereoisomers, as well as isolated specific stereoisomers, are also included. During the course of the synthetic procedures used to prepare such compounds, or in using racemization or epimerization procedures known to those skilled in the art, the products of such procedures can be a mixture of stereoisomers.

[0101] Many organic compounds exist in optically active forms having the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are non-superimposable mirror images of one another. A specific stereoisomer can also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture. Many of the compounds described herein can have one or more chiral centers and therefore can exist in different enantiomeric forms. If desired, a chiral carbon can be designated with an asterisk (*). When bonds to the chiral carbon are depicted as straight lines in the disclosed formulas, it is understood that both the (R) and (S) configurations of the chiral carbon, and hence both enantiomers and mixtures thereof, are embraced within the formula. As is used in the art, when it is desired to specify the absolute configuration about a chiral carbon, one of the bonds to the chiral carbon can be depicted as a wedge (bonds to atoms above the plane) and the other can be depicted as aATTORNEY DOCKET NO.222112-2430 series or wedge of short parallel lines is (bonds to atoms below the plane). The Cahn-Inglod- Prelog system can be used to assign the (R) or (S) configuration to a chiral carbon.

[0102] Compounds described herein comprise atoms in both their natural isotopic abundance and in non-natural abundance. The disclosed compounds can be isotopically-labeled or isotopically-substituted compounds identical to those described, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of isotopes that can be incorporated into compounds of the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine and chlorine, such as2H,3H,13C,14C,15N,18O,17O,35S,18F, and36Cl, respectively. Compounds further comprise prodrugs thereof and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of this disclosure. Certain isotopically- labeled compounds of the present disclosure, for example those into which radioactive isotopes such as3H and14C are incorporated, are useful in drug and / or substrate tissue distribution assays. Tritiated, i.e.,3H, and carbon-14, i.e.,14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e.,2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labeled compounds of the present disclosure and prodrugs thereof can generally be prepared by carrying out the procedures below, by substituting a readily available isotopically labeled reagent for a non- isotopically labeled reagent.

[0103] Certain materials, compounds, compositions, and components disclosed herein can be obtained commercially or readily synthesized using techniques generally known to those of skill in the art. For example, the starting materials and reagents used in preparing the disclosed compounds and compositions are either available from commercial suppliers such as Aldrich Chemical Co., (Milwaukee, Wis.), Acros Organics (Morris Plains, N.J.), Fisher Scientific (Pittsburgh, Pa.), or Sigma (St. Louis, Mo.) or are prepared by methods known to those skilled in the art following procedures set forth in references such as Fieser and Fieser’s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd’s Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); March’s Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition); and Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., 1989).

[0104] It is understood, that unless otherwise specified, temperatures referred to herein are based on atmospheric pressure (i.e. one atmosphere).ATTORNEY DOCKET NO.222112-2430 B. Abbreviations

[0105] AA – acrylic acid

[0106] APBA – 3-(acrylamido)phenylboronic acid

[0107] CPBA – 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid

[0108] Đ – polymer dispersity

[0109] DDMAT – 2-(dodecylthiocarbonothioylthio)-2-methylpropionic acid

[0110] DMA – N,N-dimethylacrylamide

[0111] DMF – dimethylformamide

[0112] DMSO – dimethyl sulfoxide

[0113] HEAm – 2-(hydroxyethyl)acrylamide

[0114] IBD – inflammatory bowel disease

[0115] Mn– number average molecular weight

[0116] macro-CTA – macro-chain transfer agent

[0117] NBAm – Nile blue acrylamide, N-(5-acrylamido-9H-benzo[a]phenoxazin-9-ylidene)-N- ethylethanaminium chloride

[0118] PBA – phenylboronic acid

[0119] SLIPs – Shear-Labile Interaction Polymers C. Introduction

[0120] Described herein are copolymers comprising mucoadhesive monomers that have therapeutic or clinical utility. Also described herein are pharmaceutical compositions comprising the copolymers. In one aspect, the copolymers can function as synthetic mucins. In one aspect, the copolymers can be referred to as SLIPs. The copolymers include block copolymers, gradient copolymers, and statistical / random copolymers. Also described herein are methods of synthesizing the copolymers. Also described herein are methods of administering the copolymers to a subject in need thereof. In some aspects, the subject can have a gastrointestinal condition, such as an inflammatory disease. Other compositions, compounds, methods, features, and advantages of the present disclosure will be or become apparent to one having ordinary skill in the art upon examination of the following drawings, detailed description, and examples. It is intended that all such additional compositions, compounds, methods, features, and advantages be included within this description, and be within the scope of the present disclosure. D. Mucoadhesive Polymers.

[0121] In one aspect, disclosed herein are block copolymers comprising a first block and a second block; where the first block can be selected from a mucoadhesive polymer block and a backbone polymer block; where the second block can be selected from a mucoadhesive polymer block and a backbone polymer block, provided that the first block and the second block are not the same; where the mucoadhesive polymer block comprises at least oneATTORNEY DOCKET NO.222112-2430 mucoadhesive monomer; where the backbone polymer block comprises at least one backbone monomer; and where the backbone polymer block does not contain a mucoadhesive monomer. The mucoadhesive polymer block can further include at least one backbone monomer. The block copolymer can be from about 1% to about 90%, about 1% to about 75%, about 1% to about 50%, about 1% to about 25%, or about 1% to about 10% of the mucoadhesive polymer block(s) by weight. The mucoadhesive polymer block can be comprised of monomer units, where from about 1% to about 100%, about 1% to about 90%, about 1% to about 70%, about 10% to about 100%, about 10% to about 90%, about 10% to about 70%, or about 30% to about 70% of the monomer units are a mucoadhesive monomer.

[0122] In another aspect, the block copolymer can include a first block comprising a mucoadhesive polymer block, a second block comprising a backbone polymer block, and a third block comprising a mucoadhesive polymer block. In a further aspect, the first and third mucoadhesive polymer blocks are the terminal blocks of the block copolymer. The first and third mucoadhesive polymer blocks can include the same mucoadhesive monomer(s), at least one different mucoadhesive monomer, or entirely different mucoadhesive monomer(s).

[0123] The block copolymer can be a triblock copolymer. In one aspect, the block copolymer can have a formula represented by the following structure: (A)x–block–(B)y–block–(A)z; where block A includes a mucoadhesive polymer block and block B includes a backbone polymer block. In another aspect, the block copolymer can have a formula represented by the following structure: (A1)x–block–(B)y–block–(A2)z; where block A1includes a first mucoadhesive polymer block; block B includes a backbone polymer block; and block A2includes a second mucoadhesive polymer block; where the first mucoadhesive polymer block comprises a different mucoadhesive monomer from the second mucoadhesive polymer block. In one aspect, the first mucoadhesive polymer block of A1can include a greater wt% of mucoadhesive monomers than the second mucoadhesive polymer block of A2. In another aspect, the first mucoadhesive polymer block of A1can bind more strongly to or have higher affinity for mucins than the second mucoadhesive polymer block of A2, e.g., A1can have a dissociation constant, Kd1, and A2can have a dissociation constant, Kd2, such that Kd1< Kd2. In a further aspect, Kd1<< Kd2. The first mucoadhesive polymer block and the second mucoadhesive polymer block can be a mucoadhesive polymer block as disclosed herein.

[0124] In the block copolymer, he mucoadhesive monomer can include at least one at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof (e.g., a derivative of a residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, or acrylamide). The backbone monomer can include at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate,ATTORNEY DOCKET NO.222112-2430 poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof (e.g., a derivative of a residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, or methacrylic acid).

[0125] In a further aspect, the first mucoadhesive polymer block of A1can include at least one mucoadhesive monomer comprising at least one residue of a boronic acid, an organoboron, or a derivative thereof. In a still further aspect, the boronic acid can be a boronic acid with a low pKa. In another further aspect, the organoboron group can be characterized as being able to strongly bind with diols. In a yet further aspect, the first mucoadhesive polymer block of A1and the second mucoadhesive polymer block of A2can include at least one mucoadhesive monomer comprising at least one residue independently selected from a boronic acid, an organoboron, or a derivative thereof. In a still further aspect, the first mucoadhesive polymer block of A1and the second mucoadhesive polymer block of A2can each comprise the same boronic acid, organoboron, or a derivative thereof. In an even further aspect, the first mucoadhesive polymer block of A1and the second mucoadhesive polymer block of A2can each comprise a different boronic acid, organoboron, or a derivative thereof.

[0126] In another aspect, disclosed herein are random / statistical copolymers comprising at least one mucoadhesive unit, where the mucoadhesive unit can include at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof (e.g., a derivative of a residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, or acrylamide); and at least one backbone unit, where the backbone unit comprises at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof (e.g., a derivative of a residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, or methacrylic acid). In the random copolymer, the mucoadhesive units are randomly distributed throughout the entirety of the copolymer. The random copolymer can be comprised of monomer units, where from about 0.01% to about 90%, about 0.01% to about 60%, about 0.01% to about 25%, about 1% to about 90%, about 1% to about 60%, about 1% to about 25%, or about 1% to about 5% of the monomer units are a mucoadhesive monomer.

[0127] In another aspect, disclosed herein are gradient copolymers comprising at least one mucoadhesive unit, where the mucoadhesive unit includes at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof (e.g., a derivative of a residue of boronicATTORNEY DOCKET NO.222112-2430 acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, or acrylamide); and at least one backbone unit, where the backbone unit comprises at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof (e.g., a derivative of a residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, or methacrylic acid). The gradient copolymer can be comprised of monomer units, where from about 0.01% to about 90%, about 0.01% to about 60%, about 0.01% to about 25%, about 1% to about 90%, about 1% to about 60%, about 1% to about 25%, or about 1% to about 5% of the monomer units are a mucoadhesive monomer. In another aspect, the gradient copolymer can include at least two mucoadhesive units, where one of the mucoadhesive units can comprise at least one mucoadhesive monomer that the other mucoadhesive units does not include.

[0128] In the gradient copolymer, there is a gradual change in composition along the copolymer from primarily mucoadhesive units near one terminal end of the polymer to primarily backbone units near the other terminal end of the polymer. More specifically, in one aspect, greater than 50% of the mucoadhesive units are contained within a terminal region of the gradient copolymer. The terminal region is defined as less than 50% of the copolymer’s overall backbone degree of polymerization. In another aspect, about 75% or about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer. In another aspect, greater than 50% to about 99% or about 75% to about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

[0129] For any of the copolymers disclosed herein (block copolymer, random copolymer, or gradient copolymer), the mucoadhesive monomer can be selected from the group of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, 4-(acrylamido)phenylboronic acid, 3- (acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 2-vinylphenylboronic acid, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, pyridyl disulfide ethyl acrylate, pyridyl disulfide ethyl acrylamide, pyridyl disulfide alkyl methacrylamide 2-(pyridin-2-yldisulfaneyl)ethyl acrylate, 2-(pyridin-2-yldisulfaneyl)ethyl acrylamide, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoy1)- 3-bromophenyl)boronic acid, and 4-((2-acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0130] In another aspect, the mucoadhesive monomer can include at least one residue of phenylboronic acid or a derivative thereof. In a further aspect, the mucoadhesive monomer can be selected from the group of 4-(acrylamido)phenylboronic acid, 3-ATTORNEY DOCKET NO.222112-2430 (acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4- ((2-acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0131] For any of the copolymers disclosed herein (block copolymer, random copolymer, or gradient copolymer), the backbone monomer can be selected from the group of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N- dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, poly(ethlylene glycol) acrylate, poly(ethylene glycol) methacrylate, 2-(3- acrylamidopropanamido)phenyl)boronic acid, and N-(2-(3-acetamido-4,5-dihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

[0132] In another aspect, the backbone monomer can include at least one residue of acrylamide, acrylic acid, or a derivative thereof. In a further aspect, the backbone monomer can be selected from the group of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, and N-(2-(3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro- 2H-pyran-2-yl)ethyl)methacrylamide.

[0133] For any of the copolymers disclosed herein (block copolymer, random copolymer, or gradient copolymer), the total molecular weight of the copolymer can range from about 10 kDa to about 50 MDa, about 10 kDa to about 10 MDa, about 100 kDa to about 50 MDa, about 100 kDa to about 10 MDa, about 0.5 MDa to about 50 MDa, or about 0.5 MDa to about 5 MDa.

[0134] In various aspects, it is contemplated herein that the disclosed compounds further comprise their biosteric equivalents. The term “bioisosteric equivalent” refers to compounds or groups that possess near equal molecular shapes and volumes, approximately the same distribution of electrons, and which exhibit similar physical and biological properties. Examples of such equivalents are: (i) fluorine vs. hydrogen, (ii) oxo vs. thia, (iii) hydroxyl vs. amide, (iv) carbonyl vs. oxime, (v) carboxylate vs. tetrazole. Examples of such bioisosteric replacements can be found in the literature and examples of such are: (i) Burger A, Relation of chemical structure and biological activity; in Medicinal Chemistry Third ed., Burger A, ed.; Wiley- Interscience; New York, 1970, 64-80; (ii) Burger, A.; “Isosterism and bioisosterism in drug design”; Prog. Drug Res.1991, 37, 287-371; (iii) Burger A, “Isosterism and bioanalogy in drug design”, Med. Chem. Res. 1994, 4, 89-92; (iv) Clark R D, Ferguson A M, Cramer R D, “Bioisosterism and molecular diversity”, Perspect. Drug Discovery Des.1998, 9 / 10 / 11, 213- 224; (v) Koyanagi T, Haga T, “Bioisosterism in agrochemicals”, ACS Symp. Ser.1995, 584,ATTORNEY DOCKET NO.222112-2430 15-24; (vi) Kubinyi H, “Molecular similarities. Part 1. Chemical structure and biological activity”, Pharm. Unserer Zeit 1998, 27, 92-106; (vii) Lipinski C A.; “Bioisosterism in drug design”; Annu. Rep. Med. Chem.1986, 21, 283-91; (viii) Patani G A, LaVoie E J, “Bioisosterism: A rational approach in drug design”, Chem. Rev. (Washington, D.C.) 1996, 96, 3147-3176; (ix) Soskic V, Joksimovic J, “Bioisosteric approach in the design of new dopaminergic / serotonergic ligands”, Curr. Med. Chem. 1998, 5, 493-512 (x) Thornber C W, “Isosterism and molecular modification in drug design”, Chem. Soc. Rev.1979, 8, 563-80.

[0135] In further aspects, bioisosteres are atoms, ions, or molecules in which the peripheral layers of electrons can be considered substantially identical. The term bioisostere is usually used to mean a portion of an overall molecule, as opposed to the entire molecule itself. Bioisosteric replacement involves using one bioisostere to replace another with the expectation of maintaining or slightly modifying the biological activity of the first bioisostere. The bioisosteres in this case are thus atoms or groups of atoms having similar size, shape and electron density. Preferred bioisosteres of esters, amides or carboxylic acids are compounds containing two sites for hydrogen bond acceptance. In one embodiment, the ester, amide or carboxylic acid bioisostere is a 5-membered monocyclic heteroaryl ring, such as an optionally substituted 1H-imidazolyl, an optionally substituted oxazolyl, 1H-tetrazolyl, [1,2,4]triazolyl, or an optionally substituted [1,2,4]oxadiazolyl.

[0136] In various aspects, it is contemplated herein that the disclosed compounds further comprise their isotopically-labelled or isotopically-substituted variants, i.e., compounds identical to those described, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as2H,3H,13C,14C,15N,18O,17O,35S,18F and36Cl, respectively. Compounds further comprise prodrugs thereof, and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labelled compounds of the present invention, for example those into which radioactive isotopes such as3H and14C are incorporated, are useful in drug and / or substrate tissue distribution assays. Tritiated, i.e.,3H, and carbon-14, i.e.,14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e.,2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labelled compounds of the present invention and prodrugs thereof can generally be prepared by carrying out the procedures below, by substituting a readily available isotopically labelled reagent for a non- isotopically labelledATTORNEY DOCKET NO.222112-2430 reagent.

[0137] In various aspects, the disclosed compounds can possess at least one center of asymmetry, they can be present in the form of their racemates, in the form of the pure enantiomers and / or diastereomers or in the form of mixtures of these enantiomers and / or diastereomers. The stereoisomers can be present in the mixtures in any arbitrary proportions. In some aspects, provided this is possible, the disclosed compounds can be present in the form of the tautomers.

[0138] Thus, methods which are known per se can be used, for example, to separate the disclosed compounds which possess one or more chiral centers and occur as racemates into their optical isomers, i.e., enantiomers or diastereomers. The separation can be effected by means of column separation on chiral phases or by means of recrystallization from an optically active solvent or using an optically active acid or base or by means of derivatizing with an optically active reagent, such as an optically active alcohol, and subsequently cleaving off the residue.

[0139] In various aspects, the disclosed compounds can be in the form of a co-crystal. The term “co-crystal” means a physical association of two or more molecules which owe their stability through non-covalent interaction. One or more components of this molecular complex provide a stable framework in the crystalline lattice. In certain instances, the guest molecules are incorporated in the crystalline lattice as anhydrates or solvates, see e.g. “Crystal Engineering of the Composition of Pharmaceutical Phases. Do Pharmaceutical Co-crystals Represent a New Path to Improved Medicines?” Almarasson, O., et. al., The Royal Society of Chemistry, 1889-1896, 2004. Preferred co-crystals include p-toluenesulfonic acid and benzenesulfonic acid.

[0140] The term “pharmaceutically acceptable co-crystal” means one that is compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0141] In a further aspect, the disclosed compounds can be isolated as solvates and, in particular, as hydrates of a disclosed compound, which can be obtained, for example, by crystallization from a solvent or from aqueous solution. In this connection, one, two, three or any arbitrary number of solvate or water molecules can combine with the compounds according to the invention to form solvates and hydrates.

[0142] The disclosed compounds can be used in the form of salts derived from inorganic or organic acids. Pharmaceutically acceptable salts include salts of acidic or basic groups present in the disclosed compounds. Suitable pharmaceutically acceptable salts include base addition salts, including alkali metal salts, e.g., sodium or potassium salts; alkaline earth metal salts, e.g., calcium or magnesium salts; and salts formed with suitable organic ligands, e.g., quaternary ammonium salts, which may be similarly prepared by reacting the drug compound with a suitable pharmaceutically acceptable base. The salts can be prepared in situ during theATTORNEY DOCKET NO.222112-2430 final isolation and purification of the compounds of the present disclosure; or following final isolation by reacting a free base function, such as a secondary or tertiary amine, of a disclosed compound with a suitable inorganic or organic acid; or reacting a free acid function, such as a carboxylic acid, of a disclosed compound with a suitable inorganic or organic base.

[0143] Acidic addition salts can be prepared in situ during the final isolation and purification of a disclosed compound, or separately by reacting moieties comprising one or more nitrogen groups with a suitable acid. In various aspects, acids which may be employed to form pharmaceutically acceptable acid addition salts include such inorganic acids as hydrochloric acid, sulphuric acid and phosphoric acid and such organic acids as oxalic acid, maleic acid, succinic acid and citric acid. In a further aspect, salts further include, but are not limited, to the following: hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzensulfonate, p- toluenesulfonate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, 2-hydroxyethanesulfonate (isethionate), nicotinate, 2-naphthalenesulfonate, oxalate, pectinate, persulfate, 3- phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, phosphate, glutamate, bicarbonate, undecanoate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3- naphthoate)) salts. Also, basic nitrogen-containing groups can be quatemized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides, and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides, and others.

[0144] Basic addition salts can be prepared in situ during the final isolation and purification of a disclosed compound, or separately by reacting carboxylic acid moieties with a suitable base such as the hydroxide, carbonate or bicarbonate of a pharmaceutical acceptable metal cation or with ammonia, or an organic primary, secondary or tertiary amine. Pharmaceutical acceptable salts include, but are not limited to, cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, aluminum salts and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Other representative organic amines useful for the formation of base addition salts include diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. In further aspects, bases which may be used in the preparation of pharmaceutically acceptable salts include the following: ammonia, L-arginine, benethamine, benzathine, calcium hydroxide, choline, deanol,ATTORNEY DOCKET NO.222112-2430 diethanolamine, diethylamine, 2-(diethylamino)-ethanol, ethanolamine, ethylenediamine, N- methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine, magnesium hydroxide, 4-(2- hydroxyethyl)-morpholine, piperazine, potassium hydroxide, 1-(2-hydroxyethyl)-pyrrolidine, secondary amine, sodium hydroxide, triethanolamine, tromethamine and zinc hydroxide. E. Methods of Making the Compounds

[0145] A method for synthesizing a polymer of the present disclosure (e.g., a block copolymer or a gradient copolymer) is as follows. In one aspect, a mixture is formed by dissolving in a solvent (e.g., DMSO): a compound comprising a thiocarbonylthio residue (−C(S)S−; e.g., DDMAT); a compound comprising a backbone residue; and, optionally, a compound comprising a mucoadhesive residue to form an initial mixture. The initial mixture can then be sparged (e.g., under argon) for less than an hour (e.g., about 5 minutes to about 30 minutes). Following sparging, the initial mixture can then be subjected to UV light (e.g., 365 nm at 4.5 mW / cm2) at about room temperature for less than a day (e.g., about 3 hours to about 10 hours) to produce an intermediate mixture comprising a first product. Optionally, the first product can be purified (e.g., by dialysis) and dried (e.g., via lyophilization).

[0146] Examples of compounds comprising a thiocarbonylthio residue include, but are not limited to, 2-(dodecylthiocarbonothioylthio)-2-methylpropionic acid, 2- (dodecylthiocarbonothioylthio)propionic acid, methyl 2-(dodecylthiocarbonothioylthio)-2- methylpropionate, 4-cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl]pentanol, 4-((((2- carboxyethyl)thio)carbonothioyl)thio)-4-cyanopentanoic acid, 2-[[(2- carboxyethyl)sulfanylthiocarbonyl]-sulfanyl]propanoic acid, 4-cyano-4- [(dodecylsulfanylthiocarbonyl)sulfanyl]pentanol, or 2-cyano-2-propyl dodecyl trithiocarbonate

[0147] The intermediate mixture can be mixed with a compound comprising a backbone residue and, optionally, a compound comprising a mucoadhesive residue to form a final mixture. Optionally, if the first product was purified and dried, the first product can be dissolved in a solvent (e.g., DMSO) along with a compound comprising a backbone residue and, optionally, a compound comprising a mucoadhesive residue to form the final mixture. The final mixture be sparged (e.g., under argon) for less than an hour (e.g., about 10 minutes to about 30 minutes) and then subjected to UV light at about room temperature for less than a day (e.g., about 8 hours to about 14 hours) to form a final crude product. The final crude product can then be purified (e.g., by dialysis) and dried (e.g., via lyophilization) to yield the final polymer.

[0148] In one aspect, in order to synthesize a gradient copolymer, the method can include a forced gradient process. In one aspect, this can refer slowly adding a compound comprising a backbone residue and, optionally, a compound comprising a mucoadhesive residue to a mixture including other components (e.g., a solvent and a compound comprising a thiocarbonylthio), rather than mixing all components together at about the same time. AATTORNEY DOCKET NO.222112-2430 method carried out according to the foregoing may utilize monomers, e.g., a compound comprising a backbone residue and, optionally, a compound comprising a mucoadhesive residue, have similar chemical reactivities. In a further aspect, in order to synthesize a gradient copolymer, the method can be carried using two different monomers comprising, e.g., a first monomer comprising a backbone residue, and a second monomer comprising a mucoadhesive residue, to a mixture including other components. In the method carried out according to the immediate foregoing, in various aspects, the first monomer and second monomer can have distinct relative chemical reactivities such that the more reactive monomer, e.g., the first monomer, is consumed preferably initially in the reaction due to a greater chemical reactivity. Thereby, the polymer with a higher concentration of the residue derived from the first monomer and the other end has a lower relative concentration of the first monomer because the first monomer is being depleted during the initial phases of polymerization leading to the distal end of the polymer having a greater relative concentration of residues derived from the second polymer.

[0149] In another aspect, polymers of the present disclosure (e.g., a block copolymer, a gradient copolymer, or a random copolymer) can be formed using various radical polymerization approaches, such conventional free radical polymerization, photoiniferter polymerization, reversible-addition fragmentation chain transfer polymerization, macromolecular design by interchange of xanthate polymerization, and atom-transfer radical polymerization. F. Pharmaceutical Compositions

[0150] In various aspects, the present disclosure relates to pharmaceutical compositions comprising a therapeutically effective amount of at least one disclosed compound (e.g., a block copolymer, a random copolymer, or a gradient copolymer as described herein), at least one product of a disclosed method, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. As used herein, “pharmaceutically-acceptable carriers” means one or more of a pharmaceutically acceptable diluents, preservatives, antioxidants, solubilizers, emulsifiers, coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, and adjuvants. The disclosed pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy and pharmaceutical sciences.

[0151] In a further aspect, the disclosed pharmaceutical compositions comprise a therapeutically effective amount of at least one disclosed compound, at least one product of a disclosed method, or a pharmaceutically acceptable salt thereof as an active ingredient, a pharmaceutically acceptable carrier, optionally one or more other therapeutic agent, and optionally one or more adjuvant. The disclosed pharmaceutical compositions include those suitable for oral, rectal, topical, and nasal administration, although the most suitable route inATTORNEY DOCKET NO.222112-2430 any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. In a further aspect, the disclosed pharmaceutical composition can be formulated to allow administration orally, nasally, via inhalation, paracancerally, transmucosally, intraperitonealy, intraventricularly, intracranially and intratumorally.

[0152] In various aspects, the present disclosure also relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and, as active ingredient, a therapeutically effective amount of a disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, or a stereochemically isomeric form thereof. In a further aspect, a disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, or a stereochemically isomeric form thereof, or any subgroup or combination thereof may be formulated into various pharmaceutical forms for administration purposes.

[0153] Pharmaceutically acceptable salts can be prepared from pharmaceutically acceptable non-toxic bases or acids. For therapeutic use, salts of the disclosed compounds are those wherein the counter ion is pharmaceutically acceptable. However, salts of acids and bases which are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound. All salts, whether pharmaceutically acceptable or not, are contemplated by the present disclosure. Pharmaceutically acceptable acid and base addition salts are meant to comprise the therapeutically active non-toxic acid and base addition salt forms which the disclosed compounds are able to form.

[0154] In various aspects, a disclosed compound comprising an acidic group or moiety, e.g., a carboxylic acid group, can be used to prepare a pharmaceutically acceptable salt. For example, such a disclosed compound may comprise an isolation step comprising treatment with a suitable inorganic or organic base. In some cases, it may be desirable in practice to initially isolate a compound from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert the latter back to the free acid compound by treatment with an acidic reagent, and subsequently convert the free acid to a pharmaceutically acceptable base addition salt. These base addition salts can be readily prepared using conventional techniques, e.g., by treating the corresponding acidic compounds with an aqueous solution containing the desired pharmacologically acceptable cations and then evaporating the resulting solution to dryness, preferably under reduced pressure. Alternatively, they also can be prepared by mixing lower alkanolic solutions of the acidic compounds and the desired alkali metal alkoxide together, and then evaporating the resulting solution to dryness in the same manner as before.

[0155] Bases which can be used to prepare the pharmaceutically acceptable base-additionATTORNEY DOCKET NO.222112-2430 salts of the base compounds are those which can form non-toxic base-addition salts, i.e., salts containing pharmacologically acceptable cations such as, alkali metal cations (e.g., lithium, potassium and sodium), alkaline earth metal cations (e.g., calcium and magnesium), ammonium or other water-soluble amine addition salts such as N-methylglucamine- (meglumine), lower alkanolammonium and other such bases of organic amines. In a further aspect, derived from pharmaceutically acceptable organic non-toxic bases include primary, secondary, and tertiary amines, as well as cyclic amines and substituted amines such as naturally occurring and synthesized substituted amines. In various aspects, such pharmaceutically acceptable organic non-toxic bases include, but are not limited to, ammonia, methylamine, ethylamine, propylamine, isopropylamine, any of the four butylamine isomers, betaine, caffeine, choline, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, N,N'-dibenzylethylenediamine, pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, tromethamine, 2- diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, quinuclidine, pyridine, quinoline and isoquinoline; benzathine, N-methyl-D-glucamine, ethylenediamine, N-ethylmorpholine, N- ethylpiperidine, glucamine, glucosamine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, hydrabamine salts, and salts with amino acids such as, for example, histidine, arginine, lysine and the like. The foregoing salt forms can be converted by treatment with acid back into the free acid form.

[0156] In various aspects, a disclosed compound comprising a protonatable group or moiety, e.g., an amino group, can be used to prepare a pharmaceutically acceptable salt. For example, such a disclosed compound may comprise an isolation step comprising treatment with a suitable inorganic or organic acid. In some cases, it may be desirable in practice to initially isolate a compound from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert the latter back to the free base compound by treatment with an basic reagent, and subsequently convert the free base to a pharmaceutically acceptable acid addition salt. These acid addition salts can be readily prepared using conventional techniques, e.g., by treating the corresponding basic compounds with an aqueous solution containing the desired pharmacologically acceptable anions and then evaporating the resulting solution to dryness, preferably under reduced pressure. Alternatively, they also can be prepared by treating the free base form of the disclosed compound with a suitable pharmaceutically acceptable non-toxic inorganic or organic acid.

[0157] Acids which can be used to prepare the pharmaceutically acceptable acid-addition salts of the base compounds are those which can form non-toxic acid-addition salts, i.e., salts containing pharmacologically acceptable anions formed from their corresponding inorganic and organic acids. Exemplary, but non-limiting, inorganic acids include hydrochloric hydrobromic, sulfuric, nitric, phosphoric and the like. Exemplary, but non-limiting, organicATTORNEY DOCKET NO.222112-2430 acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, isethionic, lactic, maleic, malic, mandelicmethanesulfonic, mucic, pamoic, pantothenic, succinic, tartaric, p-toluenesulfonic acid and the like. In a further aspect, the acid-addition salt comprises an anion formed from hydrobromic, hydrochloric, maleic, phosphoric, sulfuric, and tartaric acids.

[0158] In practice, the compounds of the present disclosure, or pharmaceutically acceptable salts thereof, of the present disclosure can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral. Thus, the pharmaceutical compositions of the present disclosure can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the compounds of the present disclosure, and / or pharmaceutically acceptable salt(s) thereof, can also be administered by controlled release means and / or delivery devices. The compositions can be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.

[0159] It is especially advantageous to formulate the aforementioned pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. The term “unit dosage form,” as used herein, refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. That is, a “unit dosage form” is taken to mean a single dose wherein all active and inactive ingredients are combined in a suitable system, such that the patient or person administering the drug to the patient can open a single container or package with the entire dose contained therein, and does not have to mix any components together from two or more containers or packages. Typical examples of unit dosage forms are tablets (including scored or coated tablets), capsules or pills for oral administration; single dose vials for injectable solutions or suspension; suppositories for rectal administration; powder packets; wafers; and segregated multiples thereof. This list of unit dosage forms is not intended to be limiting in any way, but merely to represent typical examples of unit dosage forms.

[0160] The pharmaceutical compositions disclosed herein comprise a compound of theATTORNEY DOCKET NO.222112-2430 present disclosure (or pharmaceutically acceptable salts thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally one or more additional therapeutic agents. In various aspects, the disclosed pharmaceutical compositions can include a pharmaceutically acceptable carrier and a disclosed compound, or a pharmaceutically acceptable salt thereof. In a further aspect, a disclosed compound, or pharmaceutically acceptable salt thereof, can also be included in a pharmaceutical composition in combination with one or more other therapeutically active compounds. The instant compositions include compositions suitable for oral, rectal, and topical (including subcutaneous and intramuscular) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions can be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.

[0161] Techniques and compositions for making dosage forms useful for materials and methods described herein are described, for example, in the following references: Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al., 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976); Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol 7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate Carriers: Therapeutic Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G. Wilson, Eds.); Modern Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.).

[0162] The compounds described herein are typically to be administered in admixture with suitable pharmaceutical diluents, excipients, extenders, or carriers (termed herein as a pharmaceutically acceptable carrier, or a carrier) suitably selected with respect to the intended form of administration and as consistent with conventional pharmaceutical practices. The deliverable compound will be in a form suitable for oral, rectal, or topical administration. Carriers include solids or liquids, and the type of carrier is chosen based on the type of administration being used. The compounds may be administered as a dosage that has a known quantity of the compound.

[0163] Because of the ease in administration, oral administration can be a preferred dosage form, and tablets and capsules represent the most advantageous oral dosage unit forms inATTORNEY DOCKET NO.222112-2430 which case solid pharmaceutical carriers are obviously employed. However, other dosage forms may be suitable depending upon clinical population (e.g., age and severity of clinical condition), solubility properties of the specific disclosed compound used, and the like. Accordingly, the disclosed compounds can be used in oral dosage forms such as pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. In preparing the compositions for oral dosage form, any convenient pharmaceutical media can be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like can be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets can be coated by standard aqueous or nonaqueous techniques.

[0164] The disclosed pharmaceutical compositions in an oral dosage form can comprise one or more pharmaceutical excipient and / or additive. Non-limiting examples of suitable excipients and additives include gelatin, natural sugars such as raw sugar or lactose, lecithin, pectin, starches (for example corn starch or amylose), dextran, polyvinyl pyrrolidone, polyvinyl acetate, gum arabic, alginic acid, tylose, talcum, lycopodium, silica gel (for example colloidal), cellulose, cellulose derivatives (for example cellulose ethers in which the cellulose hydroxy groups are partially etherified with lower saturated aliphatic alcohols and / or lower saturated, aliphatic oxyalcohols, for example methyl oxypropyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose phthalate), fatty acids as well as magnesium, calcium or aluminum salts of fatty acids with 12 to 22 carbon atoms, in particular saturated (for example stearates), emulsifiers, oils and fats, in particular vegetable (for example, peanut oil, castor oil, olive oil, sesame oil, cottonseed oil, corn oil, wheat germ oil, sunflower seed oil, cod liver oil, in each case also optionally hydrated); glycerol esters and polyglycerol esters of saturated fatty acids C12H24O2to C18H36O2and their mixtures, it being possible for the glycerol hydroxy groups to be totally or also only partly esterified (for example mono-, di- and triglycerides); pharmaceutically acceptable mono- or multivalent alcohols and polyglycols such as polyethylene glycol and derivatives thereof, esters of aliphatic saturated or unsaturated fatty acids (2 to 22 carbon atoms, in particular 10-18 carbon atoms) with monovalent aliphatic alcohols (1 to 20 carbon atoms) or multivalent alcohols such as glycols, glycerol, diethylene glycol, pentacrythritol, sorbitol, mannitol and the like, which may optionally also be etherified, esters of citric acid with primary alcohols, acetic acid, urea, benzyl benzoate, dioxolanes, glyceroformals, tetrahydrofurfuryl alcohol, polyglycol ethers with C1-C12-alcohols, dimethylacetamide, lactamides, lactates, ethylcarbonates, silicones (in particular medium-ATTORNEY DOCKET NO.222112-2430 viscous polydimethyl siloxanes), calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate, magnesium carbonate and the like.

[0165] Other auxiliary substances useful in preparing an oral dosage form are those which cause disintegration (so-called disintegrants), such as: cross-linked polyvinyl pyrrolidone, sodium carboxymethyl starch, sodium carboxymethyl cellulose or microcrystalline cellulose. Conventional coating substances may also be used to produce the oral dosage form. Those that may for example be considered are: polymerizates as well as copolymerizates of acrylic acid and / or methacrylic acid and / or their esters; copolymerizates of acrylic and methacrylic acid esters with a lower ammonium group content (for example EudragitR RS), copolymerizates of acrylic and methacrylic acid esters and trimethyl ammonium methacrylate (for example EudragitR RL); polyvinyl acetate; fats, oils, waxes, fatty alcohols; hydroxypropyl methyl cellulose phthalate or acetate succinate; cellulose acetate phthalate, starch acetate phthalate as well as polyvinyl acetate phthalate, carboxy methyl cellulose; methyl cellulose phthalate, methyl cellulose succinate, -phthalate succinate as well as methyl cellulose phthalic acid half ester; zein; ethyl cellulose as well as ethyl cellulose succinate; shellac, gluten; ethylcarboxyethyl cellulose; ethacrylate-maleic acid anhydride copolymer; maleic acid anhydride-vinyl methyl ether copolymer; styrol-maleic acid copolymerizate; 2-ethyl-hexyl- acrylate maleic acid anhydride; crotonic acid-vinyl acetate copolymer; glutaminic acid / glutamic acid ester copolymer; carboxymethylethylcellulose glycerol monooctanoate; cellulose acetate succinate; polyarginine.

[0166] Plasticizing agents that may be considered as coating substances in the disclosed oral dosage forms are: citric and tartaric acid esters (acetyl-triethyl citrate, acetyl tributyl-, tributyl- , triethyl-citrate); glycerol and glycerol esters (glycerol diacetate, -triacetate, acetylated monoglycerides, castor oil); phthalic acid esters (dibutyl-, diamyl-, diethyl-, dimethyl-, dipropyl- phthalate), di-(2-methoxy- or 2-ethoxyethyl)-phthalate, ethylphthalyl glycolate, butylphthalylethyl glycolate and butylglycolate; alcohols (propylene glycol, polyethylene glycol of various chain lengths), adipates (diethyladipate, di-(2-methoxy- or 2-ethoxyethyl)-adipate; benzophenone; diethyl- and diburylsebacate, dibutylsuccinate, dibutyltartrate; diethylene glycol dipropionate; ethyleneglycol diacetate, -dibutyrate, -dipropionate; tributyl phosphate, tributyrin; polyethylene glycol sorbitan monooleate (polysorbates such as Polysorbar 50); sorbitan monooleate.

[0167] Moreover, suitable binders, lubricants, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents may be included as carriers. The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include, but are not limited to, lactose, terra alba, sucrose, glucose, methylcellulose, dicalcium phosphate, calcium sulfate, mannitol, sorbitol talc, starch, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup,ATTORNEY DOCKET NO.222112-2430 peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0168] In various aspects, a binder can include, for example, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. In a further aspect, a disintegrator can include, for example, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

[0169] In various aspects, an oral dosage form, such as a solid dosage form, can comprise a disclosed compound that is attached to polymers as targetable drug carriers or as a prodrug. Suitable biodegradable polymers useful in achieving controlled release of a drug include, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, caprolactones, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and hydrogels, preferably covalently crosslinked hydrogels.

[0170] Tablets may contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.

[0171] A tablet containing a disclosed compound can be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets can be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent.

[0172] In various aspects, a solid oral dosage form, such as a tablet, can be coated with an enteric coating to prevent ready decomposition in the stomach. In various aspects, enteric coating agents include, but are not limited to, hydroxypropylmethylcellulose phthalate, methacrylic acid-methacrylic acid ester copolymer, polyvinyl acetate-phthalate and cellulose acetate phthalate. Akihiko Hasegawa “Application of solid dispersions of Nifedipine with enteric coating agent to prepare a sustained-release dosage form” Chem. Pharm. Bull. 33:1615-1619 (1985). Various enteric coating materials may be selected on the basis of testing to achieve an enteric coated dosage form designed ab initio to have a preferableATTORNEY DOCKET NO.222112-2430 combination of dissolution time, coating thicknesses and diametral crushing strength (e.g., see S. C. Porter et al. “The Properties of Enteric Tablet Coatings Made From Polyvinyl Acetate- phthalate and Cellulose acetate Phthalate”, J. Pharm. Pharmacol.22:42p (1970)). In a further aspect, the enteric coating may comprise hydroxypropyl-methylcellulose phthalate, methacrylic acid-methacrylic acid ester copolymer, polyvinyl acetate-phthalate and cellulose acetate phthalate.

[0173] In various aspects, an oral dosage form can be a solid dispersion with a water soluble or a water insoluble carrier. Examples of water soluble or water insoluble carrier include, but are not limited to, polyethylene glycol, polyvinylpyrrolidone, hydroxypropylmethyl-cellulose, phosphatidylcholine, polyoxyethylene hydrogenated castor oil, hydroxypropylmethylcellulose phthalate, carboxymethylethylcellulose, or hydroxypropylmethylcellulose, ethyl cellulose, or stearic acid.

[0174] In various aspects, an oral dosage form can be in a liquid dosage form, including those that are ingested, or alternatively, administered as a mouth wash or gargle. For example, a liquid dosage form can include aqueous suspensions, which contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. In addition, oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may also contain various excipients. The pharmaceutical compositions of the present disclosure may also be in the form of oil-in-water emulsions, which may also contain excipients such as sweetening and flavoring agents.

[0175] For the preparation of solutions or suspensions it is, for example, possible to use water, particularly sterile water, or physiologically acceptable organic solvents, such as alcohols (ethanol, propanol, isopropanol, 1,2-propylene glycol, polyglycols and their derivatives, fatty alcohols, partial esters of glycerol), oils (for example peanut oil, olive oil, sesame oil, almond oil, sunflower oil, soya bean oil, castor oil, bovine hoof oil), paraffins, dimethyl sulphoxide, triglycerides and the like.

[0176] In the case of a liquid dosage form such as a drinkable solutions, the following substances may be used as stabilizers or solubilizers: lower aliphatic mono- and multivalent alcohols with 2-4 carbon atoms, such as ethanol, n-propanol, glycerol, polyethylene glycols with molecular weights between 200-600 (for example 1 to 40% aqueous solution), diethylene glycol monoethyl ether, 1,2-propylene glycol, organic amides, for example amides of aliphatic C1-C6-carboxylic acids with ammonia or primary, secondary or tertiary C1-C4-amines or C1- C4-hydroxy amines such as urea, urethane, acetamide, N-methyl acetamide, N,N-diethyl acetamide, N,N-dimethyl acetamide, lower aliphatic amines and diamines with 2-6 carbon atoms, such as ethylene diamine, hydroxyethyl theophylline, tromethamine (for example as 0.1 to 20% aqueous solution), aliphatic amino acids.ATTORNEY DOCKET NO.222112-2430

[0177] In preparing the disclosed liquid dosage form can comprise solubilizers and emulsifiers such as the following non-limiting examples can be used: polyvinyl pyrrolidone, sorbitan fatty acid esters such as sorbitan trioleate, phosphatides such as lecithin, acacia, tragacanth, polyoxyethylated sorbitan monooleate and other ethoxylated fatty acid esters of sorbitan, polyoxyethylated fats, polyoxyethylated oleotriglycerides, linolizated oleotriglycerides, polyethylene oxide condensation products of fatty alcohols, alkylphenols or fatty acids or also 1-methyl-3-(2-hydroxyethyl)imidazolidone-(2). In this context, polyoxyethylated means that the substances in question contain polyoxyethylene chains, the degree of polymerization of which generally lies between 2 and 40 and in particular between 10 and 20. Polyoxyethylated substances of this kind may for example be obtained by reaction of hydroxyl group-containing compounds (for example mono- or diglycerides or unsaturated compounds such as those containing oleic acid radicals) with ethylene oxide (for example 40 Mol ethylene oxide per 1 Mol glyceride). Examples of oleotriglycerides are olive oil, peanut oil, castor oil, sesame oil, cottonseed oil, corn oil. See also Dr. H. P. Fiedler “Lexikon der Hillsstoffe für Pharmazie, Kostnetik und angrenzende Gebiete” 1971, pages 191-195.

[0178] In various aspects, a liquid dosage form can further comprise preservatives, stabilizers, buffer substances, flavor correcting agents, sweeteners, colorants, antioxidants and complex formers and the like. Complex formers which may be for example be considered are: chelate formers such as ethylene diamine retrascetic acid, nitrilotriacetic acid, diethylene triamine pentacetic acid and their salts.

[0179] It may optionally be necessary to stabilize a liquid dosage form with physiologically acceptable bases or buffers to a pH range of approximately 6 to 9. Preference may be given to as neutral or weakly basic a pH value as possible (up to pH 8).

[0180] In order to enhance the solubility and / or the stability of a disclosed compound in a disclosed liquid dosage form, it can be advantageous to employ α-, β- or γ-cyclodextrins or their derivatives, in particular hydroxyalkyl substituted cyclodextrins, e.g.2-hydroxypropyl-β- cyclodextrin or sulfobutyl-β-cyclodextrin. Also co-solvents such as alcohols may improve the solubility and / or the stability of the compounds according to the present disclosure in pharmaceutical compositions.

[0181] In various aspects, a disclosed liquid dosage form can further comprise liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.

[0182] Pharmaceutical compositions of the present disclosure can be in a form suitable for topical administration. As used herein, the phrase “topical application” means administration onto a biological surface, whereby the biological surface includes, for example, a skin area (e.g., hands, forearms, elbows, legs, face, nails, anus and genital areas) or a mucosalATTORNEY DOCKET NO.222112-2430 membrane. By selecting the appropriate carrier and optionally other ingredients that can be included in the composition, as is detailed herein below, the compositions of the present invention may be formulated into any form typically employed for topical application. A topical pharmaceutical composition can be in a form of a cream, an ointment, a paste, a gel, a lotion, milk, a suspension, an aerosol, a spray, foam, a dusting powder, a pad, and a patch. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations can be prepared, utilizing a compound of the present disclosure, or pharmaceutically acceptable salts thereof, via conventional processing methods. As an example, a cream or ointment is prepared by mixing hydrophilic material and water, together with about 5 wt% to about 10 wt% of the compound, to produce a cream or ointment having a desired consistency.

[0183] Ointments are semisolid preparations, typically based on petrolatum or petroleum derivatives. The specific ointment base to be used is one that provides for optimum delivery for the active agent chosen for a given formulation, and, preferably, provides for other desired characteristics as well (e.g., emollience). As with other carriers or vehicles, an ointment base should be inert, stable, nonirritating and nonsensitizing. As explained in Remington: The Science and Practice of Pharmacy, 19th Ed., Easton, Pa.: Mack Publishing Co. (1995), pp. 1399-1404, ointment bases may be grouped in four classes: oleaginous bases; emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginous ointment bases include, for example, vegetable oils, fats obtained from animals, and semisolid hydrocarbons obtained from petroleum. Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin and hydrophilic petrolatum. Emulsion ointment bases are either water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions, and include, for example, cetyl alcohol, glyceryl monostearate, lanolin and stearic acid. Preferred water-soluble ointment bases are prepared from polyethylene glycols of varying molecular weight.

[0184] Lotions are preparations that are to be applied to the skin surface without friction. Lotions are typically liquid or semiliquid preparations in which solid particles, including the active agent, are present in a water or alcohol base. Lotions are typically preferred for treating large body areas, due to the ease of applying a more fluid composition. Lotions are typically suspensions of solids, and oftentimes comprise a liquid oily emulsion of the oil-in-water type. It is generally necessary that the insoluble matter in a lotion be finely divided. Lotions typically contain suspending agents to produce better dispersions as well as compounds useful for localizing and holding the active agent in contact with the skin, such as methylcellulose, sodium carboxymethyl-cellulose, and the like.

[0185] Creams are viscous liquids or semisolid emulsions, either oil-in-water or water-in-oil. Cream bases are typically water-washable, and contain an oil phase, an emulsifier and an aqueous phase. The oil phase, also called the “internal” phase, is generally comprised ofATTORNEY DOCKET NO.222112-2430 petrolatum and / or a fatty alcohol such as cetyl or stearyl alcohol. The aqueous phase typically, although not necessarily, exceeds the oil phase in volume, and generally contains a humectant. The emulsifier in a cream formulation is generally a nonionic, anionic, cationic or amphoteric surfactant. Reference may be made to Remington: The Science and Practice of Pharmacy, supra, for further information.

[0186] Pastes are semisolid dosage forms in which the bioactive agent is suspended in a suitable base. Depending on the nature of the base, pastes are divided between fatty pastes or those made from a single-phase aqueous gel. The base in a fatty paste is generally petrolatum, hydrophilic petrolatum and the like. The pastes made from single-phase aqueous gels generally incorporate carboxymethylcellulose or the like as a base. Additional reference may be made to Remington: The Science and Practice of Pharmacy, for further information.

[0187] Gel formulations are semisolid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially uniformly throughout the carrier liquid, which is typically aqueous, but also, preferably, contain an alcohol and, optionally, an oil. Preferred organic macromolecules, i.e., gelling agents, are crosslinked acrylic acid polymers such as the family of carbomer polymers, e.g., carboxypolyalkylenes that may be obtained commercially under the trademark Carbopol™. Other types of preferred polymers in this context are hydrophilic polymers such as polyethylene oxides, polyoxyethylene- polyoxypropylene copolymers and polyvinylalcohol; modified cellulose, such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. In order to prepare a uniform gel, dispersing agents such as alcohol or glycerin can be added, or the gelling agent can be dispersed by trituration, mechanical mixing or stirring, or combinations thereof.

[0188] Sprays generally provide the active agent in an aqueous and / or alcoholic solution which can be misted onto the skin for delivery. Such sprays include those formulated to provide for concentration of the active agent solution at the site of administration following delivery, e.g., the spray solution can be primarily composed of alcohol or other like volatile liquid in which the active agent can be dissolved. Upon delivery to the skin, the carrier evaporates, leaving concentrated active agent at the site of administration.

[0189] Foam compositions are typically formulated in a single or multiple phase liquid form and housed in a suitable container, optionally together with a propellant which facilitates the expulsion of the composition from the container, thus transforming it into a foam upon application. Other foam forming techniques include, for example the “Bag-in-a-can” formulation technique. Compositions thus formulated typically contain a low-boiling hydrocarbon, e.g., isopropane. Application and agitation of such a composition at the body temperature cause the isopropane to vaporize and generate the foam, in a manner similar toATTORNEY DOCKET NO.222112-2430 a pressurized aerosol foaming system. Foams can be water-based or aqueous alkanolic, but are typically formulated with high alcohol content which, upon application to the skin of a user, quickly evaporates, driving the active ingredient through the upper skin layers to the site of treatment.

[0190] Skin patches typically comprise a backing, to which a reservoir containing the active agent is attached. The reservoir can be, for example, a pad in which the active agent or composition is dispersed or soaked, or a liquid reservoir. Patches typically further include a frontal water permeable adhesive, which adheres and secures the device to the treated region. Silicone rubbers with self-adhesiveness can alternatively be used. In both cases, a protective permeable layer can be used to protect the adhesive side of the patch prior to its use. Skin patches may further comprise a removable cover, which serves for protecting it upon storage.

[0191] Examples of patch configuration which can be utilized with the present invention include a single-layer or multi-layer drug-in-adhesive systems which are characterized by the inclusion of the drug directly within the skin-contacting adhesive. In such a transdermal patch design, the adhesive not only serves to affix the patch to the skin, but also serves as the formulation foundation, containing the drug and all the excipients under a single backing film. In the multi-layer drug-in-adhesive patch a membrane is disposed between two distinct drug- in-adhesive layers or multiple drug-in-adhesive layers are incorporated under a single backing film.

[0192] Examples of pharmaceutically acceptable carriers that are suitable for pharmaceutical compositions for topical applications include carrier materials that are well-known for use in the cosmetic and medical arts as bases for e.g., emulsions, creams, aqueous solutions, oils, ointments, pastes, gels, lotions, milks, foams, suspensions, aerosols and the like, depending on the final form of the composition. Representative examples of suitable carriers according to the present invention therefore include, without limitation, water, liquid alcohols, liquid glycols, liquid polyalkylene glycols, liquid esters, liquid amides, liquid protein hydrolysates, liquid alkylated protein hydrolysates, liquid lanolin and lanolin derivatives, and like materials commonly employed in cosmetic and medicinal compositions. Other suitable carriers according to the present invention include, without limitation, alcohols, such as, for example, monohydric and polyhydric alcohols, e.g., ethanol, isopropanol, glycerol, sorbitol, 2- methoxyethanol, diethyleneglycol, ethylene glycol, hexyleneglycol, mannitol, and propylene glycol; ethers such as diethyl or dipropyl ether; polyethylene glycols and methoxypolyoxyethylenes (carbowaxes having molecular weight ranging from 200 to 20,000); polyoxyethylene glycerols, polyoxyethylene sorbitols, stearoyl diacetin, and the like.

[0193] Topical compositions of the present disclosure can, if desired, be presented in a pack or dispenser device, such as an FDA-approved kit, which may contain one or more unit dosage forms containing the active ingredient. The dispenser device may, for example, comprise aATTORNEY DOCKET NO.222112-2430 tube. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser device may also be accompanied by a notice in a form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the compositions for human or veterinary administration. Such notice, for example, may include labeling approved by the U.S. Food and Drug Administration for prescription drugs or of an approved product insert. Compositions comprising the topical composition of the invention formulated in a pharmaceutically acceptable carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0194] Another patch system configuration which can be used by the present invention is a reservoir transdermal system design which is characterized by the inclusion of a liquid compartment containing a drug solution or suspension separated from the release liner by a semi-permeable membrane and adhesive. The adhesive component of this patch system can either be incorporated as a continuous layer between the membrane and the release liner or in a concentric configuration around the membrane. Yet another patch system configuration which can be utilized by the present invention is a matrix system design which is characterized by the inclusion of a semisolid matrix containing a drug solution or suspension which is in direct contact with the release liner. The component responsible for skin adhesion is incorporated in an overlay and forms a concentric configuration around the semisolid matrix.

[0195] Pharmaceutical compositions of the present disclosure can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories can be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.

[0196] Pharmaceutical compositions containing a compound of the present disclosure, and / or pharmaceutically acceptable salts thereof, can also be prepared in powder or liquid concentrate form.

[0197] The pharmaceutical composition (or formulation) may be packaged in a variety of ways. Generally, an article for distribution includes a container that contains the pharmaceutical composition in an appropriate form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, foil blister packs, and the like. The container may also include a tamper proof assemblage to prevent indiscreet access to the contents of the package. In addition, the container typically has deposited thereon a label that describes the contents of the container and any appropriate warnings or instructions.

[0198] The disclosed pharmaceutical compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the activeATTORNEY DOCKET NO.222112-2430 ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. Pharmaceutical compositions comprising a disclosed compound formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0199] The exact dosage and frequency of administration depends on the particular disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, solvate, or polymorph thereof, a hydrate thereof, a solvate thereof, a polymorph thereof, or a stereochemically isomeric form thereof; the particular condition being treated and the severity of the condition being treated; various factors specific to the medical history of the subject to whom the dosage is administered such as the age; weight, sex, extent of disorder and general physical condition of the particular subject, as well as other medication the individual may be taking; as is well known to those skilled in the art. Furthermore, it is evident that said effective daily amount may be lowered or increased depending on the response of the treated subject and / or depending on the evaluation of the physician prescribing the compounds of the present disclosure.

[0200] Depending on the mode of administration, the pharmaceutical composition will comprise from 0.05 to 99 % by weight, preferably from 0.1 to 70 % by weight, more preferably from 0.1 to 50 % by weight of the active ingredient, and, from 1 to 99.95 % by weight, preferably from 30 to 99.9 % by weight, more preferably from 50 to 99.9 % by weight of a pharmaceutically acceptable carrier, all percentages being based on the total weight of the composition.

[0201] In the treatment conditions associated with depleted mucus activity an appropriate dosage level will generally be about 0.01 to 1000 mg per kg patient body weight per day and can be administered in single or multiple doses. In various aspects, the dosage level will be about 0.1 to about 500 mg / kg per day, about 0.1 to 250 mg / kg per day, or about 0.5 to 100 mg / kg per day. A suitable dosage level can be about 0.01 to 1000 mg / kg per day, about 0.01 to 500 mg / kg per day, about 0.01 to 250 mg / kg per day, about 0.05 to 100 mg / kg per day, or about 0.1 to 50 mg / kg per day. Within this range the dosage can be 0.05 to 0.5, 0.5 to 5.0 or 5.0 to 50 mg / kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 mg of the active ingredient, particularly 1.0, 5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900 and 1000 mg of theATTORNEY DOCKET NO.222112-2430 active ingredient for the symptomatic adjustment of the dosage of the patient to be treated. The compound can be administered on a regimen of 1 to 4 times per day, preferably once or twice per day. This dosing regimen can be adjusted to provide the optimal therapeutic response.

[0202] Such unit doses as described hereinabove and hereinafter can be administered more than once a day, for example, 2, 3, 4, 5 or 6 times a day. In various aspects, such unit doses can be administered 1 or 2 times per day, so that the total dosage for a 70 kg adult is in the range of 0.001 to about 15 mg per kg weight of subject per administration. In a further aspect, dosage is 0.01 to about 1.5 mg per kg weight of subject per administration, and such therapy can extend for a number of weeks or months, and in some cases, years. It will be understood, however, that the specific dose level for any particular patient will depend on a variety of factors including the activity of the specific compound employed; the age, body weight, general health, sex and diet of the individual being treated; the time and route of administration; the rate of excretion; other drugs that have previously been administered; and the severity of the particular disease undergoing therapy, as is well understood by those of skill in the area.

[0203] A typical dosage can be one 1 mg to about 100 mg tablet or 1 mg to about 300 mg taken once a day, or, multiple times per day, or one time-release capsule or tablet taken once a day and containing a proportionally higher content of active ingredient. The time-release effect can be obtained by capsule materials that dissolve at different pH values, by capsules that release slowly by osmotic pressure, or by any other known means of controlled release.

[0204] It can be necessary to use dosages outside these ranges in some cases as will be apparent to those skilled in the art. Further, it is noted that the clinician or treating physician will know how and when to start, interrupt, adjust, or terminate therapy in conjunction with individual patient response.

[0205] The present disclosure is further directed to a method for the manufacture of a medicament for enhancing a mucus layer (e.g., treatment of one or more mucosal disorders associated with mucosal barrier or mucosal membrane dysfunction) in mammals (e.g., humans) comprising combining one or more disclosed compounds, products, or compositions with a pharmaceutically acceptable carrier or diluent. Thus, in one aspect, the present disclosure further relates to a method for manufacturing a medicament comprising combining at least one disclosed compound or at least one disclosed product with a pharmaceutically acceptable carrier or diluent.

[0206] The disclosed pharmaceutical compositions can further comprise other therapeutically active compounds, which are usually applied in the treatment of the above mentioned pathological or clinical conditions.

[0207] It is understood that the disclosed compositions can be prepared from the disclosed compounds. It is also understood that the disclosed compositions can be employed in theATTORNEY DOCKET NO.222112-2430 disclosed methods of using.

[0208] As already mentioned, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, and a pharmaceutically acceptable carrier. Additionally, the present disclosure relates to a process for preparing such a pharmaceutical composition, characterized in that a pharmaceutically acceptable carrier is intimately mixed with a therapeutically effective amount of a compound according to the present disclosure.

[0209] As already mentioned, the present disclosure also relates to a pharmaceutical composition comprising a disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, and one or more other drugs in the treatment, prevention, control, amelioration, or reduction of risk of diseases or conditions for a disclosed compound or the other drugs may have utility as well as to the use of such a composition for the manufacture of a medicament. The present disclosure also relates to a combination of disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, or a polymorph thereof. The present disclosure also relates to such a combination for use as a medicine. The present disclosure also relates to a product comprising (a) disclosed compound, a product of a disclosed method of making, a pharmaceutically acceptable salt, a hydrate thereof, a solvate thereof, a polymorph thereof, and (b) an additional gastrointestinal therapeutic agent, as a combined preparation for simultaneous, separate or sequential use in the treatment or prevention of a condition in a mammal, including a human, the treatment or prevention of which is affected or facilitated by the modulatory effect of the disclosed compound and the additional therapeutic agent. The different drugs of such a combination or product may be combined in a single preparation together with pharmaceutically acceptable carriers or diluents, or they may each be present in a separate preparation together with pharmaceutically acceptable carriers or diluents. G. Methods of Using the Compounds

[0210] In a further aspect, the present disclosure provides methods of treatment comprising administration of a therapeutically effective amount of a disclosed compound or pharmaceutical composition as disclosed herein above to a subject in need thereof. In particular, the disclosed compounds and disclosed pharmaceutical compositions can be used in methods of treating a disease, disorder, or condition associated with dysfunction in a mucosal membrane (e.g., mucus depletion in a mucus layer in contact with epithelial cells). The disclosed compounds and disclosed pharmaceutical compositions can be used to enhance or restore a mucosal barrier to provide a clinical or therapeutic benefit to a subject which has been determined to or been diagnosed to have a condition associated with aATTORNEY DOCKET NO.222112-2430 depleted mucus layer in contact with epithelial cells, such as a gastrointestinal condition.

[0211] In some aspects of the disclosed method, the subject has been diagnosed with a disorder treatable by enhancing or restoring a mucosal barrier prior to the administering step. In some aspects of the disclosed method, the subject has been diagnosed with a gastrointestinal condition, an inflammatory disease (e.g., inflammatory bowel disease), colorectal cancer, or an enteric infection. In some aspects of the disclosed methods, the subject has been identified with a need for treatment prior to the administering step.

[0212] The disclosed compounds can be used as single agents or in combination with one or more other drugs in the treatment, prevention, control, amelioration or reduction of risk of the aforementioned diseases, disorders and conditions for which compounds of or the other drugs have utility, where the combination of drugs together are safer or more effective than either drug alone. The other drug(s) can be administered by a route and in an amount commonly used therefore, contemporaneously or sequentially with a disclosed compound. When a disclosed compound is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the disclosed compound is preferred. However, the combination therapy can also be administered on overlapping schedules. It is also envisioned that the combination of one or more active ingredients and a disclosed compound will be more efficacious than as a single agent.

[0213] The disclosed compounds can be used in combination with one or more other drugs in the treatment, prevention, control, amelioration or reduction of risk of inflammatory bowel disease for which compounds of or the other drugs have utility, where the combination of drugs together are safer or more effective than either drug alone. Agents known to treat inflammatory bowel disease include anti-inflammatory drugs, immune system suppressors, antibiotics, biologics, and the like. In a further aspect, an agent known to treat inflammatory bowel disease can include mesalamine, balsalazide, olsalazine, azathioprine, mercaptopurine, methotrexate, tofacitinib, upadacitinib, ozanimod, infliximab, adalimumab, golimumab, certolizumab, vedolizumab, ustekinumab, risankizumab, ciprofloxacin, metronidazole, or combinations thereof.

[0214] Gut barrier dysfunction plays a role in developing and progressing inflammatory diseases such as inflammatory bowel disease (IBD). The gut barrier refers to the lining of the gastrointestinal tract, which acts as a protective barrier between the intestinal contents and the underlying tissues. When this barrier becomes compromised or "leaky," it can have several implications for IBD. For instance, a leaky gut allows the passage of harmful substances, such as bacteria and toxins, from the intestinal lumen into the underlying tissues. These substances trigger an immune response and inflammation, contributing to the pathogenesis of IBD and leading to chronic inflammation. Gut barrier dysfunction can also disrupt the balance of the gut microbiota, allowing for the overgrowth of harmful bacteria such as Enterobacteriaceae,ATTORNEY DOCKET NO.222112-2430 E. coli, and Fusobacterium, and a decrease in beneficial bacteria (Firmicutes, Bacteroidetes, Lactobacillus, and Eubacterium), leading to dysbiosis. The dysbiotic state can perpetuate inflammation, impair immune regulation, and contribute to the progression of IBD.

[0215] Addressing gut barrier dysfunction is one therapeutic target in IBD management. Improving gut barrier function and reducing intestinal permeability may help mitigate inflammation, promote mucosal healing, and prevent disease progression. These strategies may include using medications, such as anti-inflammatory agents and immunomodulators, as well as dietary modifications, probiotics, and prebiotics to restore the balance of the gut microbiota. Mucus depletion leaves mucosal membranes, especially in the gut, at risk of a weakened defense against infections, increased inflammation, and damaged epithelial tissue.

[0216] Accordingly, in various aspects, the present disclosure pertains to methods of treating a variety of diseases or disorders, including, but not limited to, gastrointestinal conditions, autoimmune diseases, immune and inflammatory diseases, cancers such as colorectal cancer, and infectious diseases. H. Kits.

[0217] In a further aspect, the present disclosure relates to kits comprising at least one disclosed compound, or a pharmaceutically acceptable salt, hydrate, solvate, or polymorph thereof, and one or more of: (a) at least one agent known to treat a gastrointestinal condition; (b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane; (c) instructions for treating a gastrointestinal condition; (d) instructions for treating a disorder associated with mucus depletion in a mucosal membrane.

[0218] The disclosed compounds and / or pharmaceutical compositions comprising the disclosed compounds can conveniently be presented as a kit, whereby two or more components, which may be active or inactive ingredients, carriers, diluents, and the like, are provided with instructions for preparation of the actual dosage form by the patient or person administering the drug to the patient. Such kits may be provided with all necessary materials and ingredients contained therein, or they may contain instructions for using or making materials or components that must be obtained independently by the patient or person administering the drug to the patient. In further aspects, a kit can include optional components that aid in the administration of the unit dose to patients, such as vials for reconstituting powder forms, syringes for injection, customized IV delivery systems, inhalers, etc. Additionally, a kit can contain instructions for preparation and administration of the compositions. The kit can be manufactured as a single use unit dose for one patient, multiple uses for a particular patient (at a constant dose or in which the individual compounds may vary in potency as therapy progresses); or the kit may contain multiple doses suitable for administration to multiple patients (“bulk packaging”). The kit components may be assembled in cartons, blister packs, bottles, tubes, and the like.ATTORNEY DOCKET NO.222112-2430

[0219] In a further aspect, the disclosed kits can be packaged in a daily dosing regimen (e.g., packaged on cards, packaged with dosing cards, packaged on blisters or blow-molded plastics, etc.). Such packaging promotes products and increases patient compliance with drug regimens. Such packaging can also reduce patient confusion. The present invention also features such kits further containing instructions for use.

[0220] In a further aspect, the present disclosure also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention. Associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.

[0221] In various aspects, the disclosed kits can also comprise compounds and / or products co-packaged, co-formulated, and / or co-delivered with other components. For example, a drug manufacturer, a drug reseller, a physician, a compounding shop, or a pharmacist can provide a kit comprising a disclosed compound and / or product and another component for delivery to a patient.

[0222] It is contemplated that the disclosed kits can be used in connection with the disclosed methods of making, the disclosed methods of using or treating, and / or the disclosed compositions. I. Aspects

[0223] Aspect 1. A pharmaceutical composition comprising a therapeutically effective amount of at least one block copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the block copolymer comprises a first block and a second block; wherein the first block is selected from a mucoadhesive polymer block and a backbone polymer block; wherein the second block is selected from a mucoadhesive polymer block and a backbone polymer block, provided that the first block and the second block are not the same; wherein the mucoadhesive polymer block comprises at least one mucoadhesive monomer; wherein the backbone polymer block comprises at least one backbone monomer; wherein the backbone polymer block does not contain a mucoadhesive monomer; wherein the mucoadhesive monomer comprises at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and wherein the backbone monomer comprises at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof.ATTORNEY DOCKET NO.222112-2430

[0224] Aspect 2. The pharmaceutical composition of aspect 1, wherein the mucoadhesive monomer is selected from the group consisting of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, 4-(acrylamido)phenylboronic acid, 3-(acrylamido)phenylboronic acid, 2- (acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 2- vinylphenylboronic acid, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, pyridyl disulfide ethyl acrylate, pyridyl disulfide ethyl acrylamide, pyridyl disulfide alkyl methacrylamide 2-(pyridin-2- yldisulfaneyl)ethyl acrylate, 2-(pyridin-2-yldisulfaneyl)ethyl acrylamide, 2-(pyridin-2- yldisulfaneyl)ethyl methacrylate, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 4-((2- acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoyl)- 3-fluorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2-acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0225] Aspect 3. The pharmaceutical composition of aspect 1, wherein the mucoadhesive monomer is selected from the group consisting of 4-(acrylamido)phenylboronic acid, 3- (acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4- ((2-acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0226] Aspect 4. The pharmaceutical composition of any one of aspects 1-3, wherein the mucoadhesive monomer comprises at least one residue of phenylboronic acid or a derivative thereof.

[0227] Aspect 5. The pharmaceutical composition of any one of aspects 1-4, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N- alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, poly(ethlylene glycol) acrylate, poly(ethylene glycol) methacrylate, 2-(3-acrylamidopropanamido)phenyl)boronic acid, and N- (2-(3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)ethyl)methacrylamide.

[0228] Aspect 6. The pharmaceutical composition of any one of aspects 1-4, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N- alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, and N-(2-(3-acetamido-4,5- dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

[0229] Aspect 7. The pharmaceutical composition of any one of aspects 1-4, wherein the backbone monomer is selected from acrylic acid and N,N-dimethylacrylamide.ATTORNEY DOCKET NO.222112-2430

[0230] Aspect 8. The pharmaceutical composition of any one of aspects 1-7, wherein the backbone monomer comprises at least one residue of acrylamide, acrylic acid, or a derivative thereof.

[0231] Aspect 9. The pharmaceutical composition of any one of aspects 1-8, wherein the mucoadhesive polymer block further comprises at least one backbone monomer.

[0232] Aspect 10. The pharmaceutical composition of any one of aspects 1-9, wherein the block copolymer further comprises a third block; wherein the block copolymer comprises a first block comprising a first mucoadhesive polymer block and a third block comprising a second mucoadhesive polymer block; and wherein the first mucoadhesive polymer block and the second mucoadhesive polymer block are terminal polymer blocks.

[0233] Aspect 11. The pharmaceutical composition of aspect 10, wherein the first block comprises the same mucoadhesive monomer as the third block.

[0234] Aspect 12. The pharmaceutical composition of aspect 10, wherein the first block comprises a different mucoadhesive monomer from the third block.

[0235] Aspect 13. The pharmaceutical composition of any one of aspects 1-10, wherein the block copolymer has a formula represented by the following structure: (A)x–block–(B)y–block– (A)z; wherein block A is the first block comprising a mucoadhesive polymer block; and wherein block B is the second block comprising a backbone polymer block.

[0236] Aspect 14. The pharmaceutical composition of any one of aspects 1-10, wherein the block copolymer has a formula represented by the following structure: (A1)x–block–(B)y–block– (A2)z; wherein block A1is the first block comprising a first mucoadhesive polymer block; wherein block B is the second block comprising a backbone polymer block; wherein block A2is the third block comprising a second mucoadhesive polymer block; and wherein the first block comprises a different mucoadhesive monomer from the third block.

[0237] Aspect 15. The pharmaceutical composition of aspect 14, wherein the first block comprises a greater wt% of mucoadhesive monomers than the third block.

[0238] Aspect 16. The pharmaceutical composition of aspect 14, wherein the first block comprises at least one mucoadhesive monomer comprising at least one residue of a boronic acid, an organoboron, or a derivative thereof.

[0239] Aspect 17. The pharmaceutical composition of any one of aspects 1-16, wherein the total molecular weight of the block copolymer ranges from about 10 kDa to about 50 MDa.

[0240] Aspect 18. The pharmaceutical composition of any one of aspects 1-16, wherein the total molecular weight of the block copolymer ranges from about 100 kDa to about 10 MDa.

[0241] Aspect 19. The pharmaceutical composition of any one of aspects 1-16, wherein the total molecular weight of the block copolymer ranges from about 0.5 MDa to about 5 MDa.ATTORNEY DOCKET NO.222112-2430

[0242] Aspect 20. The pharmaceutical composition of any one of aspects 1-19, wherein the block copolymer is from about 1% to about 90% of the at least one mucoadhesive polymer block by weight.

[0243] Aspect 21. The pharmaceutical composition of any one of aspects 1-19, wherein the block copolymer is from about 1% to about 25% of the at least one mucoadhesive polymer block by weight.

[0244] Aspect 22. The block copolymer of any one of aspects 1-19, wherein the block copolymer is from about 1% to about 10% of the at least one mucoadhesive polymer block by weight.

[0245] Aspect 23. The pharmaceutical composition of any one of aspects 1-22, wherein the mucoadhesive polymer block is comprised of monomer units, wherein from about 1% to about 100% of the monomer units are the mucoadhesive monomer.

[0246] Aspect 24. The pharmaceutical composition of any one of aspects 1-22, wherein the mucoadhesive polymer block is comprised of monomer units, wherein from about 10% to about 90% of the monomer units are the mucoadhesive monomer.

[0247] Aspect 25. The pharmaceutical composition of any one of aspects 1-22, wherein the mucoadhesive polymer block is comprised of monomer units, wherein from about 30% to about 70% of the monomer units are the mucoadhesive monomer.

[0248] Aspect 26. The pharmaceutical composition of any one of aspects 1-25, further comprising at least one agent known to treat a gastrointestinal condition.

[0249] Aspect 27. The pharmaceutical composition of aspect 26, wherein the gastrointestinal condition is an inflammatory disease.

[0250] Aspect 28. The pharmaceutical composition of aspect 27, wherein the inflammatory disease is an inflammatory bowel disease.

[0251] Aspect 29. The pharmaceutical composition of aspect 28, wherein the at least one agent known to treat the inflammatory bowel disease is an anti-inflammatory drug, an immune system suppressor, an antibiotic, a biologic, or other agent known to treat the inflammatory bowel disease.

[0252] Aspect 30. The pharmaceutical composition of aspect 28 or aspect 29, wherein the at least one agent known to treat the inflammatory bowel disease is selected from the group consisting of mesalamine, balsalazide, olsalazine, azathioprine, mercaptopurine, methotrexate, tofacitinib, upadacitinib, ozanimod, infliximab, adalimumab, golimumab, certolizumab, vedolizumab, ustekinumab, risankizumab, ciprofloxacin, metronidazole, or combinations thereof.

[0253] Aspect 31. The pharmaceutical composition of aspect 26, wherein the gastrointestinal condition is colorectal cancer.ATTORNEY DOCKET NO.222112-2430

[0254] Aspect 32. The pharmaceutical composition of aspect 26, wherein the gastrointestinal condition is an enteric infection.

[0255] Aspect 33. A pharmaceutical composition comprising a therapeutically effective amount of at least one random copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the random copolymer comprises: at least one mucoadhesive unit, wherein the mucoadhesive unit comprises at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and at least one backbone unit, wherein the backbone unit comprises at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof. wherein the mucoadhesive units are randomly distributed throughout the random copolymer.

[0256] Aspect 34. The pharmaceutical composition of aspect 33, wherein the mucoadhesive monomer is selected from the group consisting of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, 4-(acrylamido)phenylboronic acid, 3-(acrylamido)phenylboronic acid, 2- (acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 2- vinylphenylboronic acid, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, pyridyl disulfide ethyl acrylate, pyridyl disulfide ethyl acrylamide, pyridyl disulfide alkyl methacrylamide 2-(pyridin-2- yldisulfaneyl)ethyl acrylate, 2-(pyridin-2-yldisulfaneyl)ethyl acrylamide, 2-(pyridin-2- yldisulfaneyl)ethyl methacrylate, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 4-((2- acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoyl)- 3-fluorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2-acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0257] Aspect 35. The pharmaceutical composition of aspect 33, wherein the mucoadhesive monomer is selected from the group consisting of 4-(acrylamido)phenylboronic acid, 3- (acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4- ((2-acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0258] Aspect 36. The pharmaceutical composition of any one of aspects 33-35, wherein the mucoadhesive monomer comprises at least one residue of phenylboronic acid or a derivative thereof.

[0259] Aspect 37. The pharmaceutical composition of any one of aspects 33-36, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid,ATTORNEY DOCKET NO.222112-2430 acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N- alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, poly(ethlylene glycol) acrylate, poly(ethylene glycol) methacrylate, 2-(3-acrylamidopropanamido)phenyl)boronic acid, and N- (2-(3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)ethyl)methacrylamide.

[0260] Aspect 38. The pharmaceutical composition of any one of aspects 33-36, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N- alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, and N-(2-(3-acetamido-4,5- dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

[0261] Aspect 39. The pharmaceutical composition of any one of aspects 33-36, wherein the backbone monomer is selected from acrylic acid and N,N-dimethylacrylamide.

[0262] Aspect 40. The pharmaceutical composition of any one of aspects 33-39, wherein the backbone monomer comprises at least one residue of acrylamide, acrylic acid, or a derivative thereof.

[0263] Aspect 41. The pharmaceutical composition of any one of aspects 33-40, wherein the mucoadhesive unit further comprises at least one backbone monomer.

[0264] Aspect 42. The pharmaceutical composition of any one of aspects 33-41, wherein the random copolymer comprises at least two mucoadhesive units, wherein one of the mucoadhesive units comprises at least one mucoadhesive monomer that the other mucoadhesive unit does not.

[0265] Aspect 43. The pharmaceutical composition of any one of aspects 33-42, wherein the total molecular weight of the random copolymer ranges from about 10 kDa to about 50 MDa.

[0266] Aspect 44. The pharmaceutical composition of any one of aspects 33-42, wherein the total molecular weight of the random copolymer ranges from about 100 kDa to about 10 MDa.

[0267] Aspect 45. The pharmaceutical composition of any one of aspects 33-42, wherein the total molecular weight of the random copolymer ranges from about 0.5 MDa to about 5 MDa.

[0268] Aspect 46. The pharmaceutical composition of any one of aspects 33-45, wherein the random copolymer is comprised of monomer units, wherein from about 0.01% to about 90% of the monomer units are the mucoadhesive unit.

[0269] Aspect 47. The pharmaceutical composition of any one of aspects 33-45, wherein the random copolymer is comprised of monomer units, wherein from about 0.01% to about 60% of the monomer units are the mucoadhesive unit.ATTORNEY DOCKET NO.222112-2430

[0270] Aspect 48. The pharmaceutical composition of any one of aspects 33-45, wherein the random copolymer is comprised of monomer units, wherein from about 1% to about 25% of the monomer units are the mucoadhesive unit.

[0271] Aspect 49. The pharmaceutical composition of any one of aspects 33-45, wherein the random copolymer is comprised of monomer units, wherein from about 1% to about 5% of the monomer units are the mucoadhesive unit.

[0272] Aspect 50. The pharmaceutical composition of any one of aspects 33-49, further comprising at least one agent known to treat a gastrointestinal condition.

[0273] Aspect 51. The pharmaceutical composition of aspect 50, wherein the gastrointestinal condition is an inflammatory disease.

[0274] Aspect 52. The pharmaceutical composition of aspect 51, wherein the inflammatory disease is an inflammatory bowel disease.

[0275] Aspect 53. The pharmaceutical composition of aspect 52, wherein the at least one agent known to treat the inflammatory bowel disease is an anti-inflammatory drug, an immune system suppressor, an antibiotic, a biologic, or other agent known to treat the inflammatory bowel disease.

[0276] Aspect 54. The pharmaceutical composition of aspect 52 or aspect 53, wherein the at least one agent known to treat the inflammatory bowel disease is selected from the group consisting of mesalamine, balsalazide, olsalazine, azathioprine, mercaptopurine, methotrexate, tofacitinib, upadacitinib, ozanimod, infliximab, adalimumab, golimumab, certolizumab, vedolizumab, ustekinumab, risankizumab, ciprofloxacin, metronidazole, or combinations thereof.

[0277] Aspect 55. The pharmaceutical composition of aspect 50, wherein the gastrointestinal condition is colorectal cancer.

[0278] Aspect 56. The pharmaceutical composition of aspect 50, wherein the gastrointestinal condition is an enteric infection.

[0279] Aspect 57. A pharmaceutical composition comprising a therapeutically effective amount of at least one gradient copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the gradient copolymer comprises: at least one mucoadhesive unit, wherein the mucoadhesive unit comprises at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and at least one backbone unit, wherein the backbone unit comprises at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylicATTORNEY DOCKET NO.222112-2430 acid, methacrylic acid, or a derivative thereof; wherein greater than 50% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

[0280] Aspect 58. The pharmaceutical composition of aspect 57, wherein about 75% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

[0281] Aspect 59. The pharmaceutical composition of aspect 57, wherein about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

[0282] Aspect 60. The pharmaceutical composition of aspect 57, wherein greater than 50% to about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

[0283] Aspect 61. The pharmaceutical composition of aspect 57, wherein about 75% to about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

[0284] Aspect 62. The pharmaceutical composition of any one of aspects 57-61, wherein the mucoadhesive monomer is selected from the group consisting of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, 4-(acrylamido)phenylboronic acid, 3-(acrylamido)phenylboronic acid, 2- (acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 2- vinylphenylboronic acid, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, pyridyl disulfide ethyl acrylate, pyridyl disulfide ethyl acrylamide, pyridyl disulfide alkyl methacrylamide 2-(pyridin-2- yldisulfaneyl)ethyl acrylate, 2-(pyridin-2-yldisulfaneyl)ethyl acrylamide, 2-(pyridin-2- yldisulfaneyl)ethyl methacrylate, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 4-((2- acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoyl)- 3-fluorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2-acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0285] Aspect 63. The pharmaceutical composition of any one of aspects 57-61, wherein the mucoadhesive monomer is selected from the group consisting of 4-(acrylamido)phenylboronic acid, 3-(acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4- vinylphenylboronic acid, 3-vinylphenylboronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3- chlorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4- ((2-acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

[0286] Aspect 64. The pharmaceutical composition of any one of aspects 57-63, wherein the mucoadhesive monomer comprises at least one residue of phenylboronic acid or a derivative thereof.

[0287] Aspect 65. The pharmaceutical composition of any one of aspects 57-64, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N- alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethyleneATTORNEY DOCKET NO.222112-2430 glycol) acrylamide, poly(ethylene glycol) methacrylamide, poly(ethlylene glycol) acrylate, poly(ethylene glycol) methacrylate, 2-(3-acrylamidopropanamido)phenyl)boronic acid, and N- (2-(3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)ethyl)methacrylamide.

[0288] Aspect 66. The pharmaceutical composition of any one of aspects 57-64, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N- alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, and N-(2-(3-acetamido-4,5- dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

[0289] Aspect 67. The pharmaceutical composition of any one of aspects 57-64, wherein the backbone monomer is selected from acrylic acid and N,N-dimethylacrylamide.

[0290] Aspect 68. The pharmaceutical composition of any one of aspects 57-67, wherein the backbone monomer comprises at least one residue of acrylamide, acrylic acid, or a derivative thereof.

[0291] Aspect 69. The pharmaceutical composition of any one of aspects 57-68, wherein the mucoadhesive unit further comprises at least one backbone monomer.

[0292] Aspect 70. The pharmaceutical composition of any one of aspects 57-69, wherein the gradient copolymer comprises at least two mucoadhesive units, wherein one of the mucoadhesive units comprises at least one mucoadhesive monomer that the other mucoadhesive unit does not.

[0293] Aspect 71. The pharmaceutical composition of any one of aspects 57-70, wherein the total molecular weight of the gradient copolymer ranges from about 10 kDa to about 50 MDa.

[0294] Aspect 72. The pharmaceutical composition of any one of aspects 57-70, wherein the total molecular weight of the gradient copolymer ranges from about 100 kDa to about 10 MDa.

[0295] Aspect 73. The pharmaceutical composition of any one of aspects 57-70, wherein the total molecular weight of the gradient copolymer ranges from about 0.5 MDa to about 5 MDa.

[0296] Aspect 74. The pharmaceutical composition of any one of aspects 57-73, wherein the gradient copolymer is comprised of monomer units, wherein from about 0.01% to about 90% of the monomer units are the mucoadhesive unit.

[0297] Aspect 75. The pharmaceutical composition of any one of aspects 57-73, wherein the gradient copolymer is comprised of monomer units, wherein from about 0.01% to about 60% of the monomer units are the mucoadhesive unit.

[0298] Aspect 76. The pharmaceutical composition of any one of aspects 57-73, wherein the gradient copolymer is comprised of monomer units, wherein from about 1% to about 25% of the monomer units are the mucoadhesive unit.ATTORNEY DOCKET NO.222112-2430

[0299] Aspect 77. The pharmaceutical composition of any one of aspects 57-73, wherein the gradient copolymer is comprised of monomer units, wherein from about 1% to about 5% of the monomer units are the mucoadhesive unit.

[0300] Aspect 78. The pharmaceutical composition of any one of aspects 57-77, further comprising at least one agent known to treat a gastrointestinal condition.

[0301] Aspect 79. The pharmaceutical composition of aspect 78, wherein the gastrointestinal condition is an inflammatory disease.

[0302] Aspect 80. The pharmaceutical composition of aspect 79, wherein the inflammatory disease is an inflammatory bowel disease.

[0303] Aspect 81. The pharmaceutical composition of aspect 80, wherein the at least one agent known to treat the inflammatory bowel disease is an anti-inflammatory drug, an immune system suppressor, an antibiotic, a biologic, or other agent known to treat the inflammatory bowel disease.

[0304] Aspect 82. The pharmaceutical composition of aspect 80 or aspect 81, wherein the at least one agent known to treat the inflammatory bowel disease is selected from the group consisting of mesalamine, balsalazide, olsalazine, azathioprine, mercaptopurine, methotrexate, tofacitinib, upadacitinib, ozanimod, infliximab, adalimumab, golimumab, certolizumab, vedolizumab, ustekinumab, risankizumab, ciprofloxacin, metronidazole, or combinations thereof.

[0305] Aspect 83. The pharmaceutical composition of aspect 78, wherein the gastrointestinal condition is colorectal cancer.

[0306] Aspect 84. The pharmaceutical composition of aspect 78, wherein the gastrointestinal condition is an enteric infection.

[0307] Aspect 85. A method for the treatment of a disease or disorder in a subject comprising the step of administering to the subject the pharmaceutical composition of aspect 1.

[0308] Aspect 86. The method of aspect 85, wherein the disease or disorder is associated with mucus depletion in a mucosal membrane.

[0309] Aspect 87. The method of aspect 85, wherein the disease or disorder is a gastrointestinal condition.

[0310] Aspect 88. The method of aspect 85, wherein the disease or disorder is an inflammatory disease.

[0311] Aspect 89. The method of aspect 88, wherein the inflammatory disease is an inflammatory bowel disease.

[0312] Aspect 90. The method of aspect 85, wherein the disease or disorder is colorectal cancer.

[0313] Aspect 91. The method of aspect 85, wherein the disease or disorder is an enteric infection.ATTORNEY DOCKET NO.222112-2430

[0314] Aspect 92. The method of aspect 85 or aspect 87, further comprising the step of administering a therapeutically effective amount of at least one agent known to treat a gastrointestinal condition.

[0315] Aspect 93. The method of aspect 92, wherein the pharmaceutical composition and the at least one agent are administered sequentially.

[0316] Aspect 94. The method of aspect 92, wherein the pharmaceutical composition and the at least one agent are administered simultaneously.

[0317] Aspect 95. The method of aspect 92, wherein the pharmaceutical composition and the at least one agent are co-formulated.

[0318] Aspect 96. The method of aspect 92, wherein the pharmaceutical composition and the at least one agent are co-packaged.

[0319] Aspect 97. A method for the treatment of a disease or disorder in a subject comprising the step of administering to the subject the pharmaceutical composition of aspect 33.

[0320] Aspect 98. The method of aspect 97, wherein the disease or disorder is associated with mucus depletion in a mucosal membrane.

[0321] Aspect 99. The method of aspect 97, wherein the disease or disorder is a gastrointestinal condition.

[0322] Aspect 100. The method of aspect 97, wherein the disease or disorder is an inflammatory disease.

[0323] Aspect 101. The method of aspect 100, wherein the inflammatory disease is an inflammatory bowel disease.

[0324] Aspect 102. The method of aspect 97, wherein the disease or disorder is colorectal cancer.

[0325] Aspect 103. The method of aspect 97, wherein the disease or disorder is an enteric infection.

[0326] Aspect 104. The method of aspect 97 or aspect 99, further comprising the step of administering a therapeutically effective amount of at least one agent known to treat a gastrointestinal condition.

[0327] Aspect 105. The method of aspect 104, wherein the pharmaceutical composition and the at least one agent are administered sequentially.

[0328] Aspect 106. The method of aspect 104, wherein the pharmaceutical composition and the at least one agent are administered simultaneously.

[0329] Aspect 107. The method of aspect 104, wherein the pharmaceutical composition and the at least one agent are co-formulated.

[0330] Aspect 108. The method of aspect 104, wherein the pharmaceutical composition and the at least one agent are co-packaged.ATTORNEY DOCKET NO.222112-2430

[0331] Aspect 109. A method for the treatment of a disease or disorder in a subject comprising the step of administering to the subject the pharmaceutical composition of aspect 57.

[0332] Aspect 110. The method of aspect 109, wherein the disease or disorder is associated with mucus depletion in a mucosal membrane.

[0333] Aspect 111. The method of aspect 109, wherein the disease or disorder is a gastrointestinal condition.

[0334] Aspect 112. The method of aspect 109, wherein the disease or disorder is an inflammatory disease.

[0335] Aspect 113. The method of aspect 112, wherein the inflammatory disease is an inflammatory bowel disease.

[0336] Aspect 114. The method of aspect 109, wherein the disease or disorder is colorectal cancer.

[0337] Aspect 115. The method of aspect 109, wherein the disease or disorder is an enteric infection.

[0338] Aspect 116. The method of aspect 109 or aspect 111, further comprising the step of administering a therapeutically effective amount of at least one agent known to treat a gastrointestinal condition.

[0339] Aspect 117. The method of aspect 116, wherein the pharmaceutical composition and the at least one agent are administered sequentially.

[0340] Aspect 118. The method of aspect 116, wherein the pharmaceutical composition and the at least one agent are administered simultaneously.

[0341] Aspect 119. The method of aspect 116, wherein the pharmaceutical composition and the at least one agent are co-formulated.

[0342] Aspect 120. The method of aspect 116, wherein the pharmaceutical composition and the at least one agent are co-packaged.

[0343] Aspect 121. A kit comprising the pharmaceutical composition of aspect 1 and one or more of: a) at least one agent known to treat a gastrointestinal condition; b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane; c) instructions for treating a gastrointestinal condition; or d) instructions for treating a disease or disorder associated with mucus depletion in a mucosal membrane.

[0344] Aspect 122. The kit of aspect 124, wherein the pharmaceutical composition and the at least one agent are co-formulated.

[0345] Aspect 123. The kit of aspect 124, wherein the pharmaceutical composition and the at least one agent are co-packaged.

[0346] Aspect 124. A kit comprising the pharmaceutical composition of aspect 33 and one or more of: a) at least one agent known to treat a gastrointestinal condition; b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane;ATTORNEY DOCKET NO.222112-2430 c) instructions for treating a gastrointestinal condition; or d) instructions for treating a disease or disorder associated with mucus depletion in a mucosal membrane.

[0347] Aspect 125. The kit of aspect 124, wherein the pharmaceutical composition and the at least one agent are co-formulated.

[0348] Aspect 126. The kit of aspect 124, wherein the pharmaceutical composition and the at least one agent are co-packaged.

[0349] Aspect 127. A kit comprising the pharmaceutical composition of aspect 57, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more of: a) at least one agent known to treat a gastrointestinal condition; b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane; c) instructions for treating a gastrointestinal condition; or d) instructions for treating a disease or disorder associated with mucus depletion in a mucosal membrane.

[0350] Aspect 128. The kit of aspect 127, wherein the pharmaceutical composition and the at least one agent are co-formulated.

[0351] Aspect 129. The kit of aspect 127, wherein the pharmaceutical composition and the at least one agent are co-packaged.

[0352] Aspect 130. The use of the pharmaceutical composition of aspect 1 in the manufacture of a medicament for the treatment of a disease disorder associated with mucus depletion in a mucosal membrane in a subject.

[0353] Aspect 131. The use of the pharmaceutical composition of aspect 33 in the manufacture of a medicament for the treatment of a disease disorder associated with mucus depletion in a mucosal membrane in a subject.

[0354] Aspect 132. The use of the pharmaceutical composition of aspect 57 in the manufacture of a medicament for the treatment of a disease disorder associated with mucus depletion in a mucosal membrane in a subject.

[0355] From the foregoing, it will be seen that aspects herein are well adapted to attain all the ends and objects hereinabove set forth together with other advantages which are obvious and which are inherent to the structure.

[0356] While specific elements and steps are discussed in connection to one another, it is understood that any element and / or steps provided herein is contemplated as being combinable with any other elements and / or steps regardless of explicit provision of the same while still being within the scope provided herein.

[0357] It will be understood that certain features and subcombinations are of utility and may be employed without reference to other features and subcombinations. This is contemplated by and is within the scope of the claims.

[0358] Since many possible aspects may be made without departing from the scope thereof, it is to be understood that all matter herein set forth or shown in the accompanying drawingsATTORNEY DOCKET NO.222112-2430 and detailed description is to be interpreted as illustrative and not in a limiting sense.

[0359] It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting. The skilled artisan will recognize many variants and adaptations of the aspects described herein. These variants and adaptations are intended to be included in the teachings of this disclosure and to be encompassed by the claims herein.

[0360] Now having described the aspects of the present disclosure, in general, the following Examples describe some additional aspects of the present disclosure. While aspects of the present disclosure are described in connection with the following examples and the corresponding text and figures, there is no intent to limit aspects of the present disclosure to this description. On the contrary, the intent is to cover all alternatives, modifications, and equivalents included within the spirit and scope of the present disclosure. J. Examples

[0361] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated, and are intended to be purely exemplary of the disclosure and are not intended to limit the scope of what the inventors regard as their disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in ^C or is at ambient temperature, and pressure is at or near atmospheric. 1. Synthesis and Characterization of Random and Block Copolymers

[0362] Herein are examples of synthesis schemes for random and block copolymers. The final copolymers (post-drying and purification) are characterized using H1-NMR. In preparation for NMR measurements, copolymers are dissolved in MeOD for about 2 to about 5 hours. The number average molecular weight and dispersity of the final copolymers is determined from size-exclusion chromatography-light-scattering (SEC-LS). In cases where the polymers with fluorescent tags (e.g., fluorescein, Nile Blue) have emissions that interfere with light scattering, conventional calibration from polymethylmethacrylate (PMMA) standards was used. A brief overview of the copolymers discussed herein is provided in Table 1. Table 1: Properties of Example Random Copolymers and Block Copolymers. Composition Sample Functionalization (mol%) Mn Đ % PBAATTORNEY DOCKET NO.222112-2430 DMA / AA / CPBA 81.75 DMA / 13.25 AA / 5 1.15 AT1-105 1.15 4.7% P(DMA-co-CPBA-co-AA) CPBA random MDaT1- 105, P(DMA-co-CPBA-co-AA), are represented by the structures provided in FIGS.1A and 2A, respectively. SEC-LS results along with the number average molecular weight of the copolymers and the dispersity are de presented in FIGS. 1B and 2B. H1-NMR for the copolymers are provided in FIGS.1C and 2C.

[0364] The synthesis procedure for producing AT1-104 comprises the steps of: to a 20 mL scintillation vial, DMA (1.60 mL, 15.5 mmol, 11600 equiv.), CPBA (281 mg, 0.949 mmol, 712 equiv.), AA (0.173 mL, 2.51 mmol, 1890 equiv.), and DDMAT (0.485 mg, 0.00133 mmol, 1 equiv.) were dissolved in DMSO (3.15 mL); DMF (0.3 mL) was added to the solution as an internal standard; the mixture was sparged with Ar for 15 min; and the solution was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 12 h or until full monomer conversion was observed by NMR. For this and the other copolymers discussed herein, monomer conversion was monitored via H1-NMR for decreasing vinylic protons (~5.5-6.5 ppm). The resulting polymer was purified by dialysis and dried via lyophilization.

[0365] The synthesis procedure for producing AT1-105 followed the same steps as AT1-104, with the exception of, in the first step, adding different amounts of DMA, CPBA, and AA to the scintillation vial, specifically: DMA (1.73 mL, 16.8 mmol, 12600 equiv.), CPBA (147 mg, 0.498 mmol, 373 equiv.), AA (0.181 mL, 2.64 mmol, 1980 equiv.)

[0366] Block Copolymer AT1-106: Synthesis for this diblock copolymer (Scheme 1) was a two-step, two-pot synthesis. To a 20 mL scintillation vial, DMA (0.147 mL, 1.43 mmol, 688ATTORNEY DOCKET NO.222112-2430 equiv.), CPBA (422 mg, 1.43 mmol, 688 equiv.), and DDMAT (0.755 mg, 0.00207 mmol, 1 equiv.) were dissolved in DMSO (5.66 mL). DMF (0.5 mL) was added to the solution as an internal standard. The mixture was sparged with Ar for 15 min, and the solution was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 5-8 h or until full monomer conversion was observed by NMR. The resulting macro-CTA was purified by dialysis and dried via lyophilization. Next, in a 20 mL scintillation vial, the macro-CTA (564 mg, 0.00207 mmol, 1 equiv.) was redissolved in DMSO (5.66 mL) and DMF (0.5 mL). DMA (2.65 mL, 25.7 mmol, 12400 equiv.) monomer was added to the reaction vial and the solution sparged under Ar for 15 min. The reaction was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 10-12 h or until full monomer conversion was observed by NMR. The crude mixture was purified by dialysis and dried via lyophilization to yield the final diblock copolymer.

[0367] SEC-LS results along with the number average molecular weight and dispersity of the macro-CTA and AT1-106 copolymers are presented in FIGS. 3A and 3B, respectively. H1- NMR for AT1-106 is provided in FIG.3C.ATTORNEY DOCKET NO.222112-2430 Scheme 1

[0368] Block Copolymer AT1-111: Synthesis for this diblock copolymer (Scheme 2) was a two-step, two-pot synthesis. To a 20 mL scintillation vial, DMA (0.206 mL, 2.00 mmol, 965 equiv.), CPBA (197 mg, 0.667 mmol, 322 equiv.), and DDMAT (0.755 mg, 0.00207 mmol, 1 equiv.) were dissolved in DMSO (5.86 mL). DMF (0.5 mL) was added to the solution as an internal standard. The mixture was sparged with Ar for 15 min, and the solution was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 5-8 h or until full monomer conversion was observed by NMR. The resulting macro-CTA was purified by dialysis and dried via lyophilization. Next, in a 20 mL scintillation vial, the macro-CTA (396 mg, 0.00207 mmol, 1 equiv.) was redissolved in DMSO (5.86 mL) and DMF (0.5 mL). HEAm (2.82 mL, 24.0 mmol, 11600 equiv.) monomer was added to the reaction vial and the solution sparged under Ar for 15 min. The reaction was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 10-12 h or until full monomer conversion was observed by NMR. The crude mixture was purified by dialysis and dried via lyophilization to yield the final diblock copolymer.

[0369] SEC-LS results along with the number average molecular weight and dispersity of the macro-CTA copolymer is presented in FIG.4A. The AT1-111 copolymer was insoluble, so no SEC-LS data was obtained. H1-NMR for AT1-111 is provided in FIG.4B.ATTORNEY DOCKET NO.222112-2430Scheme 2

[0370] Block Copolymer JA1-032: Synthesis for this diblock copolymer (Scheme 3) was a two- step, one-pot synthesis. To a 40 mL scintillation vial, DMA (10.7 mL, 104 mmol, 1.30 x 103equiv.), NBAm (5.99 mg, 0.0147 mmol, 11.0 equiv.), and DDMAT (2.91 mg, 0.008 mmol, 1 equiv.) were dissolved in DMSO (16.7 mL). DMF (3 mL) was added to the solution as an internal standard. The mixture was sparged with Ar for 20 min, and the solution was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 5-8 h to yield the viscous, red gel. APBA (1.10 g, 5.78 mmol, 723 equiv.) and DMA (0.596 mL, 5.78 mmol, 723 equiv.) monomer was added to the reaction vial, vortexed to homogenize, and the solution sparged under Ar for 20 min. The reaction was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 10-12 h or until full monomer conversion was observed by NMR. The crude mixture was purified by dialysis and dried via lyophilization to yield the final diblock copolymer.

[0371] SEC-LS results along with the number average molecular weight and dispersity of the JA1-032 copolymer is presented in FIG.5.ATTORNEY DOCKET NO.222112-2430Scheme 3

[0372] Block Copolymer AT1-114: Synthesis for this triblock copolymer (Scheme 4) was a two-step, two-pot synthesis. To a 20 mL scintillation vial, DMA (0.185 mL, 1.79 mmol, 775 equiv.), CPBA (531 mg, 1.79 mmol, 775 equiv.), and telechelic iniferter 2,2'- (thiocarbonylbis(sulfanediyl))bis(2-methylpropanoic acid) (0.654 mg, 0.00231 mmol, 1 equiv.) were dissolved in DMSO (6.25 mL). DMF (0.5 mL) was added to the solution as an internal standard. The mixture was sparged with Ar for 15 min, and the solution was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 5-8 h or until full monomer conversion was observed by NMR. The resulting macro-CTA was purified by dialysis and dried via lyophilization. Next, in a 20 mL scintillation vial, the macro-CTA (708 mg, 0.00231 mmol, 1 equiv.) was redissolved in DMSO (6.25 mL) and DMF (0.5 mL). DMA (2.84 mL, 27.5 mmol, 11900 equiv.) and AA (0.326 mL, 4.75 mmol, 2050 equiv.) monomers were added to the reaction vial and the solution sparged under Ar for 15 min. The reaction was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 10-12 h or until full monomer conversion was observed by NMR. The crude mixture was purified by dialysis and dried via lyophilization to yield the final triblock copolymer.

[0373] SEC-LS results along with the number average molecular weight and dispersity of AT1-114 is presented in FIG. 6A. The macro-CTA copolymer was insoluble in dimethylacetamide, so no SEC-LS data was obtained. H1-NMR for AT1-114 is provided in FIG.6B.ATTORNEY DOCKET NO.222112-2430

[0374] Block Copolymer AT1-118: Synthesis for this triblock copolymer (Scheme 5) was a two-step, two-pot synthesis. To a 20 mL scintillation vial, DMA (0.298 mL, 2.89 mmol, 362 equiv.), APBA (552 mg, 2.89 mmol, 362 equiv.), and telechelic iniferter 2,2'- (thiocarbonylbis(sulfanediyl))bis(2-methylpropanoic acid) (0.654 mg, 0.00231 mmol, 1 equiv.) were dissolved in DMSO (6.25 mL). DMF (0.5 mL) was added to the solution as an internal standard. The mixture was sparged with Ar for 15 min, and the solution was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 5-8 h or until full monomer conversion was observed by NMR. The resulting macro-CTA was purified by dialysis and dried via lyophilization. Next, in a 20 mL scintillation vial, the macro-CTA (839 mg, 0.00231 mmol, 1 equiv.) was redissolved in DMSO (6.25 mL) and DMF (0.5 mL). DMA (0.508 mL, 4.92 mmol, 617 equiv.) and AA (0.0638 mL, 0.852 mmol, 106 equiv.) monomers were added to the reaction vial and the solution sparged under Ar for 15 min. The reaction was subjected to UV light (365 nm, 4.5 mW cm2) at room temperature for 10-12 h or until full monomer conversion was observed by NMR. The crude mixture was purified by dialysis and dried via lyophilization to yield the final triblock copolymer.ATTORNEY DOCKET NO.222112-2430

[0375] AT1-118 and the macro-CTA copolymer were insoluble, so no SEC-LS data was obtained. H1-NMR for AT1-118 is provided in FIG.7. O O HOS SOH AT1-118[macroCTA] S OH2. Safety and Non-Toxicity of Synthetic Mucins

[0376] The viability of the epithelial cell line hTCEpi was determined via exposure to SLIPs for 5 days (FIG. 9A). The cells maintained high viability levels at high concentrations of 90 mg / mL. The physical barrier promoted by SLIPs was further tested in live cell imaging dishes (FIG. 9B). SLIPs were capable of interacting with mucins produced by the epithelial cells to create a robust mucus that was able to create a robust barrier between the epithelial cells and the bacterial cells, recapitulating a first line of defense against the bacteria. 3. Safety of SLIPs in Wild-Type Mice

[0377] The safety of oral doses (0, 2.5, 5, 10, 15, and 40 mg / day) was assessed in adult (12 weeks old) wild-type C57BL / 6 mice. The SLIPs contained 6.2 mol% phenylboronic acid content and a molecular weight of 1.5 MDa. Endpoints included clinical symptoms (dysphagia, consistency of feces, intestinal bleeding), body weight, rectal prolapses, gut microbiome changes, and mortality in 10 days (FIGS.10A-10F). Notably, none of the tested doses caused any adverse effects (average disease activity index (DAI) score = 0). SLIPs adhere and integrate with natural mucus, reinforcing the mucosal barrier (FIG.10B), and were constantly replenished through the drinking water. Like natural mucus, SLIPs were excreted through the feces (FIG.10D), showing that transport along the gut epithelium is the removal mechanism. SLIPs had no significant impact on the microbiome (FIG.10C, 10E, and 10F C, E, and F), butATTORNEY DOCKET NO.222112-2430 a beneficial reduction in the Firmicutes / Bacteroidetes ratio was observed in 40 mg of SLIPs per day (FIG.10E). 4. Efficacy of SLIPs in a Murine Model (Agr2- / -) Deficient in Mucin Production.

[0378] The endpoints included weight change, inflammatory response, gut permeability, and microbiome changes. The model chosen lacks exons 2-3 of the anterior gradient 2 (Agr2) gene, abolishing gene expression. Consequently, MUC2 synthesis decreases, and no mucus is produced. Eight-week-old Agr2- / -and wild-type mice received 40 mg of SLIPs daily for 10 days, and the outcomes were compared with untreated mice. Agr2- / -untreated mice displayed symptoms such as bloody stool, reduced weight, and rectal prolapse. The treatment effectively alleviated the clinical symptoms of colitis. Mice undergoing SLIPs treatment increased weight by 35% in 10 days, reaching a weight similar to that observed in their wild-type counterparts (FIG.11A). Unlike the untreated mice, SLIPs treated mice presented normal stool consistency and no blood in stools. The treatment increased fat, lean mass, and the total tissue area (FIG. 11B). SLIPs treatment also reduced the inflammatory response of Agr2- / -mice. The concentration of CXCL5, G-CSF, IFN-γ, and TNF-α significantly decreased in Agr2- / -mice treated with SLIPs (FIG.11F). In the pathogenesis of IBD, CXCL5 has been recognized as a major CXC chemokine expressed in epithelial cells of the intestinal mucosa (Refs.3 and 4). The inflammatory cytokine G-CSF is recognized as a key mediator of visceral pain in inflammatory diseases (Basso et al., 2017), and IFN-γ is a major cytokine in IBD pathogenesis (Ref.5). The Secretion of proinflammatory cytokines such as TNF-α results in tissue damage and adaptive immune system activation. Tissue damage, in turn, may result in increased exposure of the gut wall to luminal antigens, inducing a stronger activation of both the innate and adaptive immune system, perpetuating the inflammatory state and resulting in chronic inflammation. The anti-inflammatory cytokines IL-4 and IL-13 increased after treatment. To assess gut permeability, the fluorescein isothiocyanate conjugated dextran (FITC-Dextran assay 4.4-kDa) was utilized as a marker for transmucosal transport after 10 days of SLIPs treatment. The findings revealed that SLIPs treatment led to a remarkable 2.5-fold reduction in gut permeability compared to the untreated group, as illustrated in FIG.11C. Additionally, a microbiome analyses of fecal samples was performed. The results indicated that the dysbiotic outgrowth of Enterobacteriaceae tended to be reduced by SLIPs treatment (FIG.11E). When the intestinal epithelium is disrupted and inflamed, intraluminal oxygen increases, leading to the outgrowth of Enterobacteriaceae and the luminal expansion of aerobic S. Typhimurium (Refs. 6 and 7). Finally, increased microbial diversity in Agr2- / -SLIPs-treated mice was observed (FIG.11D). In IBD, reduced microbial diversity correlates with disease severity, and increased diversity is observed during remission (Refs.8 and 9). While these effects of the proposed therapy with SLIPs observed in Agr2- / -mice are not exhaustive, this initial screen provides the proof of concept for the studies proposed. The treatment effectively restored cryptATTORNEY DOCKET NO.222112-2430 structures and mitigated histological inflammation, reducing histological scores (FIG.11G and 11H). Thus, the SLIPs could reasonably be explored as an effective approach to prevent and control gut barrier dysfunction. 5. Determining the SLIPs-driven Reduction of Intestinal Permeability and Inflammatory Response

[0379] Preliminary research demonstrated the effectiveness of 40 mg of SLIPs, ingested daily for 10 days in a murine model lacking mucus synthesis. Considering the duration of the treatment and the concentration of SLIPs given to each animal, these results suggest that SLIPs should be capable of significantly reverting the expected phenotype of this model by reducing weight loss, diarrhea, blood in stool, and rectal prolapse. These findings support the need to determine intestinal permeability and inflammatory response reduction in response to treatment and identify the mechanisms by which SLIPs alleviate inflammation and restore gut health. The tendency to decrease the F / B ratio should also be investigated, as longer exposure to SLIPs may stop the leaky gut, reduce inflammation, and revert dysbiosis. However, efforts to minimize biases or limitations inherent to a particular model must be considered. Using multiple models and sex as a biological variable reduces the risk of bias, obtaining a broader perspective and minimizing the impact of model-specific biases. As such, more mechanistic data could be provided to confirm that SLIPs decrease inflammatory responses and reduce gut permeability in 3 different models.

[0380] Organoid model. Testing the effectiveness of SLIPs in reducing intestinal permeability in the human 2D epithelial barrier model using human intestinal organoids seeded on transwell dishes: It has been established that primary human organoids can be trypsinized from their 3D structures and seeded onto transwell dishes (Ref. 10). In this manner, their apical membranes face the upper chamber, and their basolateral membranes face the bottom of the membrane. Organoids from 4-6 donors will be seeded in the transwell in the presence or absence of SLIPs. Barrier permeability will be monitored by three assays: 1) The rate and ability to reach an ion-resistant monolayer will be determined by monitoring the trans-epithelial electrical resistance (TEER) of the organoids every day for seven days.2) The permeability of the monolayer will be confirmed by a dextran flow through assay, where FITC labeled dextran is added to the apical membrane and is sampled from the basolateral supernatant 3 hours post-treatment. Monolayers that have formed a tight, non-permeable layer will retain the dextran in the apical chamber, while monolayers that are damaged or leaky will allow the dextran to flow through. 3) The appearance of tight junction belts will be monitored through the staining of ZO-1 and claudin. These assays will be performed with healthy organoids from both the ileum and the colon to determine if a region-specific response to SLIPs exists. In addition, to determine if SLIPs can impact barrier formation in the presence of inflammation, organoids will be seeded in the presence of an inflammatory cocktail (TNFa and INFg) in theATTORNEY DOCKET NO.222112-2430 presence or absence of SLIPs. The barrier integrity of these organoids will be monitored by TEER, dextran flow through, and ZO-1 staining. Each of these assays will allow for an analysis SLIPs effect on the rate of monolayer formation or impact on gut permeability.

[0381] Determine to what extent SLIPs can protect the intestinal epithelial layer from the damage caused by enteric viruses and their pro-inflammatory cytokines: Enteric viruses damage the intestinal epithelium and lead to water loss and diarrhea. It has been found that enteric viruses and their pro-inflammatory cytokines can damage intestinal barriers and lead to a leaky gut. To determine if SLIPs can help protect the intestinal epithelial monolayer from this infection-induced damage, a human organoid transwell model will be employed (akin to that previously discussed). Human intestinal organoids from 4-6 healthy donors will be seeded in transwells for 7 days. Following confirmation that the monolayers have reached polarization and have formed permeable barriers (using the assay described previously), the transwells will be treated with the cytokines (IL-1β, IL-6, IL-8, TNFα) in the presence or absence of SLIPs. Previous work has shown that IL-6 alone causes the death of intestinal epithelial cells and monolayer permeability within 24 hours of treatment (Ref. 1). Following cytokine treatment, transwells will be monitored by TEER, dextran flow-through assay, and immunofluorescence of ZO-1 to determine whether SLIPs-treated samples maintain their barrier integrity better than non-treated cells. To further determine how enteric infections can impact barrier permeability, human intestinal organoids will be seeded in transwells and allowed to polarize and form an impermeable monolayer. Following monolayer formation, organoids will be treated with TLR ligands LPS and flagellin to mimic bacterial infection or will be infected with the enteric viruses (rotavirus and mammalian reovirus) in the presence or absence of SLIPs. Following TLR stimulation or virus infection, transwells will be monitored by TEER, dextran flow-through assay, and immunofluorescence of ZO-1 to determine whether SLIPs-treated samples protect the monolayer from damage induced by TLR stimulation and virus infection.

[0382] Agr2- / -model deficient in the synthesis of mucin (MUC2). Efficacy of SLIPs in reducing gut permeability and inflammatory response: The expression of mucin and TJs are dynamic and interdependent such that a decrease in one alters the other (Refs.11 and 12). In Muc2- / -mice, the structural impairment of tight junctions has been observed, as well as the delocalization of Claudin-3 (Ref.13). Intestinal barrier dysfunction results in fluid and solute leakage out of the lumen, contributing to leak-flux diarrhea and increased antigen translocation to the lamina propria (Ref. 14). Among other factors, intestinal permeability is regulated by zonula occludens toxin (Zot, or zonulin) -mediated disengagement of the protein ZO-1 from the TJ protein complex (Ref.15). Zonulin is the only known regulator of tight junctions in the gut epithelium, which is believed to increase intestinal permeability and is linked to Inflammatory Bowel Disease (Refs.16 and 17) and rheumatoid arthritis (Ref.18). The enteric mucosa not only acts as a physical sieve to exclude bacteria but contains antimicrobialATTORNEY DOCKET NO.222112-2430 peptides released by Paneth cells, such as defensins and lysozymes (Ref.19), and shares intimate communication with the innate immune system. Mucins constrain the immunogenicity of bacterial antigens and prevent inflammation by inducing dendritic cell-conditioning signals in the intestinal epithelium (Refs.2). Muc2 glycans reduce the expression of pro-inflammatory cytokines IL-6, IL-8, IL-12, and TNF-a in response to toll-like receptor (TLR) ligands such as LPS, flagellin, or TNF while enhancing expression of the anti-inflammatory cytokine IL-10 (Ref. 2). As such, Muc2 mediates gut homeostasis of resident microbiota by delivering tolerogenic signals to dendritic cells. Agr2- / -mice deficient in mucus production are predisposed to colitis and have a significant increase in the key pro-inflammatory cytokines TNF-a (7 fold) and IL- 1β (5.7 fold) (Ref.20). Preliminary results with the Agr2- / -mice showed that 40 mg of SLIPs daily for 10 days improved the body condition and reduced inflammation as measured by the DAI and inflammatory markers (FIG.3). Without intending to be bound by theory, it is possible that SLIPs reduce intestinal permeability and prevent the activation of inflammatory factors caused by mucin depletion, supporting gut homeostasis, and reducing inflammation. SLIPs will reduce the expression of pro-inflammatory cytokines IL-6, IL-8, IL-12, and TNF- α in response to toll-like receptor (TLR) ligands such as LPS, flagellin, or TNF while enhancing the expression of the anti-inflammatory cytokines.

[0383] Rigor, Reproducibility, and Sex as Biological Variable. The probe FITC-dextran, which leaks out when the gut barrier is dysregulated, will be tracked in SLIPs-treated and untreated mucin-deficient mice (Agr2- / -). The expression of tight junctions will also be measured. The protein disulfide isomerase AGR2 has already been proven essential for producing intestinal mucus (Ref.20). Based on previous data, the goal will be to collect at least 6 samples per animal group per time point in this study. In addition to p-values, effect size measures will be used to describe the results and assess the extent to which SLIPs influence the leaking gut. Animals from the same treatment will be co-housed to reduce the cage effect, and the cage will be evaluated as a confounding variable.

[0384] Specifically, Agr2- / -mice (5-7 weeks old) will be randomly assigned to receive SLIPS or not, sex-segregated, and divided into Agr2- / -treatment and Agr2- / -control groups, each comprising 6 mice. Wild-type C57BL6J mice, treated and untreated, will serve as baseline controls, bringing the total sample size to 24. Sex will be considered a biological variable randomized and balanced in the study and control groups. The treatment groups will be administered 40 mg SLIPs daily via water ad libitum for 20 days (SLIPs are well accepted as they have no taste and are charge neutral). The control groups will be given water without SLIPs in a similar manner.

[0385] Blood samples will be collected at the experiment's beginning and end. After removing the blood clot, the serum will be extracted by centrifuging at 2,000 x g for 10 minutes in a refrigerated centrifuge. To confirm the gut permeability, samples will be fractioned andATTORNEY DOCKET NO.222112-2430 assayed using enzyme-linked immunoassay (ELISA) to quantify the protein levels of the tight junction regulator Zonulin using the IDK® Zonulin (Serum) ELISA (Immundiagnostik AG, Bensheim, Germany). The standard FITC-dextran assay will be implemented to investigate intestinal permeability directly, as described previously (Ref. 21). After fasting for 4 hours, FITC-dextran will be orally gavaged, followed by euthanasia. Only a minimal amount of the 4- kDa molecule will enter the bloodstream in a normal intestinal barrier since it is nondegradable and too large to pass, therefore serving as a marker for a disrupted barrier (Refs.21 and 22). After serum dilution, the FITC fluorescence intensity will be measured using a fluorescent microplate reader. In addition, a sub-sample of the serum will be used to measure the immune response using the panel ProcartaPlex™ Mouse Th1 / Th2 Cytokine Panel, 11plex (Catalog number: EPX110-20820-901, Thermo Fisher Scientific, Carlsbad, CA) and the ProcartaPlex Mouse Cytokine Panel 1B 10plex (cat no. EPX100-26091-901, Thermo Fisher Scientific, Carlsbad, CA) with the Luminex xMAP technology. Finally, the DAI will be monitored. The DAI is the combined weight loss score compared to initial weight, stool consistency, and bleeding. Scores are defined as follows: weight loss: 0 (no loss), 1 (1-5%), 2 (5-10%), 3 (10-20%), and 4 (>20%); stool consistency: 0 (normal), 2 (loose stool), and 4 (diarrhea); and bleeding: 0 (no blood), 1 (Hemoccult positive), 2 (Hemoccult positive and visual pellet bleeding), and 4 (gross bleeding, blood around anus).

[0386] Colonic tissues from untreated and treated mice will be processed as described before (Ref. 23). Briefly, the tissue will be cleaned from adipose tissue and cut longitudinally; the luminal content will be removed by washing it in cold phosphate-buffered saline (PBS). Starting from the most distal portion (i.e., rectum) and with the luminal side facing upward, the colon will be rolled, resulting in a Swiss roll with the distal colon in the center and the proximal colon in the outer portion. The Swiss roll will be fixed in a 10% neutral buffered formalin solution, paraffin-embedded, sectioned, and H&E stained by the Molecular Pathology Core at the University of Florida. Histological scoring of inflammation will be performed blindly and calculated as the average between the proximal and distal colon region scores.

[0387] Fecal samples will be collected every other day from the start to end of the experiments, frozen promptly for microbiome analysis and SLIPs fluorescence tracking.

[0388] DSS-induced colitis model. ADSS-induced colitis model will be used to provide more mechanistic data to verify that SLIPs decrease inflammatory responses and reduce gut permeability. In the DSS model, the sulfated polysaccharide does not directly induce intestinal inflammation but acts as a direct chemical toxin to colonic epithelium, resulting in epithelial cell injury. This model is widely used and well-described in the literature (Ref. 24) and morphologically and symptomatically resembles epithelial damage seen in human ulcerative colitis.ATTORNEY DOCKET NO.222112-2430

[0389] The same rigorous experimental design described in previously will be used in this set of experiments. Wild-type C57BL / 6 mice (6 weeks old) will be randomized and separated into treatment groups to ensure unbiased distribution, with 6 mice in each group. Two different experiments will be carried out. In the first experiment, mice will receive 2.5% (w / v) DSS on days 1-3 to induce colitis and then be given 40 mg / day of SLIPs via water ad libitum for 10 days. Two control treatments will be used. The SLIP control groups will be given water with SLIPs but not treated with DSS. The DSS-control group will receive DSS but will not be treated with SLIPs. Sample collection will be done on day zero and every other day until the end of the experiment.

[0390] In a second experiment, wild-type C57BL / 6 mice (6 weeks old) will be first treated with SLIPs for 3 consecutive days (same dose described above) and then will receive 2.5% (w / v) DSS for 3 days. SLIPs will no longer be administrated, and mice will be monitored for 3 days. As described above, the control groups will be given water with SLIPs but not treated with DSS. Another control group will receive DSS but will not be treated with SLIPs. Sample collection (DAI Assessment, Stool samples) will be done after DSS treatment, SLIPs treatment, and 3 days after SLIPs treatment. Blood samples will be taken at the experiment's beginning and end. FITC-dextran assay and colon extraction will be performed at the end of the experiment.

[0391] To investigate the effect of synthetic mucin on inflammation, changes in body weight, stool consistency, and rectum bleeding will be analyzed. Data will be used to calculate the DAI. Blood samples will be used to measure pro-inflammatory activation by quantifying TNF- α, IL-1β, and IL-6 and the anti-inflammatory response through IFN-γ and IL-10 as described above. Length of the small and large intestines will be measured, and histological damage will be evaluated based on Robarts Histopathology Index scores. To confirm the gut permeability, blood samples will be used to quantify the protein levels of Zonulin, and the FITC-dextran assay will be implemented. All analyses are detailed and described above. 6. Determining how SLIPs interact with the host and the gut microbiota to reduce inflammation.

[0392] Mucus is critical in protecting and mediating the immune system. It also serves as an energy source for the gut microbiota when microbially accessible carbohydrates (MACs) are unavailable, selecting for mucolytic gut bacteria, whether beneficial such as Akkermansia muciniphila (Ref. 25) or pathogenic such as Entamoeba histolytica (Ref. 26) and other members of the Enterobacteriaceae family (Ref. 23). SLIPs, however, have an all-carbon backbone and no biological mechanism for degradation. Synthetic mucins can offer new areas for microbial colonization; however, like natural mucins, synthetic ones form a loose mucus layer that is constantly excreted and renewed (FIG. 3). This xenobiotic could prevent an accumulation of less complex sugars such as lactose, galactinol, raffinose, and melibioseATTORNEY DOCKET NO.222112-2430 contributing to the depletion of commensal Clostridia and Bacteroidetes phyla (Refs.27 and 28). As SLIPs successfully reduce the inflammatory response (FIG.3), it can also decrease dysbiosis in mucus-deficient mice potentially caused by inflammatory activation (Refs.6, 29, and 30). This interaction can activate or suppress specific immune responses, affecting the expression of inflammation-related genes, immune signaling, and defense mechanisms (Ref. 31). Therefore, understanding the mechanisms by which SLIPs treatment change gut microbial diversity and host inflammatory response is key to understanding how SLIPs reduce inflammation. Without intending to be bound by theory, it is possible that SLIPs can change the transcriptional activation of selective pro-inflammatory genes and increase gut microbial diversity, reducing enteric pathogens' survival, attachment, and growth. Molecular approaches will be used to determine the regulatory network controlling inflammation responses and microbial interactions in animal models (Agr2- / -and DSS-induced) treated with SLIPs.

[0393] Host gene expression: The mice models described in Example 5 will be used to determine the host gene expression with and without SLIPs treatment. At the endpoint, tissue samples from the colon's distal, mid, and proximal will be obtained from the mice models proposed above. After harvesting, colons will be flushed with ice-cold phosphate-buffered saline and cut open longitudinally. Samples (0.5–1^cm long) will be stored in RNAlater (Invitrogen) at -20^°C. To extract RNA, tissues will be placed in RLT Plus Buffer (Qiagen) with 2% b-mercapto-ethanol and lysed by bead-beating. RNA isolation will be performed using the RNAeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. After RNA isolation, total RNA integrity will be checked using a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). Whole RNA-seq library construction and sequencing will be performed on an Illumina NovaSeq X Plus at the UF-ICBR NextGen Sequencing (NGS) core. RNA-seq analysis will be performed as previously described (Ref. 32) and following the standard protocol for data analysis in RNA-seq experiments (Ref.33). Briefly, reads will be quality filtered at Phred score 20 and trimmed to remove remaining adapters using Trimmomatic. The resulting reads will be aligned to the Mus musculus reference genome utilizing TopHat2 (Ref.32). HTSeq2 will be used to quantify the number of reads mapped to each gene (Ref.35). Differential gene expression will be analyzed using DESeq2 (Ref.36). A gene will be considered as differentially expressed if the false discovery rate is <0.05 and the fold change is >2 for comparisons between two independent groups.

[0394] 2.1.2 Absolute Q Digital PCR validation. To verify the accuracy of the RNA-seq data and detect differentially expressed genes (DEGs) in colon samples, the most important DEGs will be selected for Absolute Q Digital PCR analysis. Genes will be selected based on DESEq2 results (FDR and log2 fold change) and their biological significance. Appropriate oligonucleotide primers will be designed using NCBI Primer-BLAST software with murine mRNA template, or the sequence will be obtained from the literature. The differentialATTORNEY DOCKET NO.222112-2430 expression will be quantified using the QuantStudio Absolute Q Digital PCR System (Applied Biosystems, USA) following the manufacturer’s instructions. The RT-dPCR reactions will be carried out in a microfluidic array plate (MAP) with Absolute Q™ 1-step RT-dPCR Master Mix (Applied Biosystems, USA) following the manufacturer’s protocol. Probes will be detected in the FAM channel. Positive control reactions will be performed as needed. Confirmation of the PCR product will be done by Sanger sequencing. Minimum Information for Publication of Quantitative Digital PCR Experiments for 2020 (Ref. 37) will be adopted. The target gene differential expression between treatments will be assessed using χ2-tests and quantified using odds ratios (ORs) with 95% confidence intervals (CIs).

[0395] Bacterial profile at the species level: This protocol has been fully optimized with high- quality control, allowing a better-quality microbial taxonomy classification at the species level. Stool samples will be collected initially and every two days until the experiment concludes. For DNA extraction, a previously described protocol will be followed (Ref.39).

[0396] All fastq files will be processed as described in (Ref.38) and the resultant contingency table with taxonomy assignments will be analyzed through the phyloseq (Ref. 40) and Microbiome (Ref.41) packages after rarefying the dataset to the minimum library size (Ref. 42). To verify whether the number of sequences obtained, where a representative of the microbial community Good's coverage, will be calculated. Possible confounding variables will be tested by Permutational Multivariate Analysis of Variance (PERMANOVA) in the vegan package (Ref.43) and controlled in the subsequent analysis. Differentially abundant species between SLIPs treatment and controls will be determined using the ALDEx2 ANOVA-Like Differential Expression tool for the compositional data (Ref. 44). Differences in microbial community structure will be observed by Principal Coordinates Analysis (PCoA) plots and tested by permutational analysis of variance (PERMANOVA) using the Vegan package with 999 permutations. As taxa counts acquired through high throughput sequencing are not absolute molecule counts, when necessary, Aitchison’s centered log-ratio transformation will be applied before calculations of all standard multivariate analysis techniques.

[0397] Fungal profile: Previous research indicates that individuals with Crohn’s Disease (CD), unlike healthy controls, exhibit elevated levels of intestinal Candida tropicalis. This fungal species is the primary one detected in the colons of mice with colitis following a dextran sodium sulfate (DSS) challenge (Refs.45 and 46). The identity and the relative abundance of microbial eukaryotes in stool samples will be determined by amplifying the ITS region using the ITS86F / ITS4 primer pair. Two hundred barcoded forward primers are available to allow 200 samples to be sequenced in a single MiSeq run. The Illumina demultiplexed sequenced dataset will be processed by the R package DADA2 to correct amplicon errors and identify chimeras and taxonomic assignments against the UNITE database. The resulting contingency table with taxonomy assignments will be analyzed as described in sub-item 2.1.ATTORNEY DOCKET NO.222112-2430

[0398] Microbial functional profile: Microbial community metabolic potential (metagenomics) will be determined by shotgun sequencing DNA from stool samples obtained on the animal experiments using the R10.4 flow cell from Oxford Nanopore on an ONT GridION sequencing instrument in the Roesch lab. Based on recent experience with these flow cells, it will be expected to obtain 30 billion bases from each flow cell. Base calling will be done by the latest version of Guppy on the ONT GridION. Choosing one flow cell per six barcoded samples taken at the beginning and end of the mice experiments can give a high level of microbial functions in each sample. The raw reads will be quality-filtered to allow no ambiguous nucleotides (N) and a maximum expected error rate of 2. The HUMAnN 3.0 will accurately profile the presence / absence and abundance of microbial pathways from sequencing data. In HUMAnN 3.0, the abundances of functions are reported by per-organism contributions rather than community-level abundances as obtained by classical approaches. The approach provides taxonomic stratification of microbial functions at the species level, thus quantifying the community abundance of functions while assigning them to specific contributors. The approach proposed by HUMAnN allows for obtaining microbial community functional profiles stratified by known and unclassified organisms, including Archaea, Bacteria, Eukaryotes, and Viruses. Differential expression between groups will be determined using ALDEx2 ANOVA- Like Differential Expression tool for compositional data. Covariates will also be investigated by PERMANOVA. 7. Synthetic Mucins for Mitigating Gut Permeability and Inflammation

[0399] The synthetic mucins (herein called SLIPs) can reduce the inflammatory response and intestinal permeability caused by mucosal disruption or inhibition of mucus secretion during pathogenesis by protecting the intestinal tract from gut microbes. Using Nano-liquid chromatography-tandem mass spectrometry (Nano-LC / MS / MS), the proteome of spleen and colon samples was quantified from Agr2- / -mice SLIPs untreated (controls) and treated with 40 mg / day of SLIPs for 17 days. From a total of 6,228 high-confidence proteins found in the spleen samples, 493 decreased after treatment. (FIGS. 12A-12D). Notably, a distinctive proteomic signature characterized by a significant decrease in the expression of proteins involved in various aspects of inflammatory signaling was observed in spleen samples of mice treated with SLIPs (FIG. 13). The main differentially expressed proteins included IL1RAP (Interleukin 1 receptor accessory protein), involved in the signaling of interleukin-1 (IL-1), a key pro-inflammatory cytokine; NOD1 (Nucleotide-binding oligomerization domain-containing protein 1), a pattern recognition receptor that detects bacterial peptidoglycan fragments. Activation of NOD1 leads to the production of pro-inflammatory cytokines and the initiation of inflammatory responses to bacterial infection; GP2 (Glycoprotein 2), involved in recognizing and clearing pathogens in the intestinal tract; NLRP10 (NLR family pyrin domain-containing 10), a NOD-like receptor (NLR) family member, which plays a critical role in inflammasomeATTORNEY DOCKET NO.222112-2430 activation and regulation. The spleen is considered a reservoir for neutrophils during colonic inflammation, being strongly involved in the immune response. In Chron's disease, it is suspected that the inflammatory cells and cytokines enter the blood vessels and affect the spleen due to the breakdown of the intestinal tract barrier. Mechanistically, restoring the epithelial barrier with SLIPs can dampen inflammatory signaling pathways, which reduce the levels of pro-inflammatory cytokines circulating in the bloodstream, thereby mitigating systemic inflammation and its effects on the spleen. The activation of immune cells within the spleen gets altered, leading to a shift towards a less inflammatory or more regulatory phenotype. Proteome analysis of colon samples revealed that SLIPs led to a significant reduction in the protein levels of Neutrophilic Granule Protein (NGP), as depicted in FIG.14 (3.71 log2Fold Change, p-value < 0.01). NGP plays a crucial role in attenuating the inflammatory response triggered by lipopolysaccharide (LPS). Therefore, the observed decrease in NGP levels suggests a concomitant reduction in LPS levels in response to the treatment. Lipopolysaccharide binding protein (LBP) and Lipocalin-2 (LCN2), two important proteins involved in immunological responses to bacterial LPS, also decreased after treatment. In total, 6081 proteins were detected in the colon samples. Among them, 815 decreased after SLIPs treatment. These findings suggest SLIPs reduce the inflammatory response and intestinal permeability caused by mucosal disruption or inhibition of mucin secretion during pathogenesis by covering and protecting the intestinal tract from gut microbes while still allowing for immunological communication. 8. In vitro Study

[0400] SLIPs Degradability. A stable, long-term solution for mucin replacement and integration with remaining natural mucins requires a material that retains functionality even in the presence of enzymatic activity. SLIPs have an all-carbon backbone and no biological mechanism for degradation. To empirically test how well SLIPs resist biological breakdown, a liquid culture media containing SLIPs as the sole carbon source was formulated. Then, it was inoculated with wild-type C57BL / 6J mice fecal pellets at various dilutions. Porcine mucin was utilized as a positive control to provide evidence that the fecal microbial community could break down natural mucins (FIG.15A). Porcine mucin (PM) (0.6 g / L) and SLIPs media (0.6 g / L) were prepared and autoclaved as described in a previous study (Ref.47). Murine fecal homogenates (100 µL) were inoculated into 1 mL of PM or SLIPs media at 1:10, 1:100, and 1:1000 dilutions, each in triplicate alongside uninoculated controls. The experiment involved five 72-hour growth cycles, incubated at 37ºC in a Mitsubishi™ AnaeroPack 7.0L Rectangular Anaerobic Jar (Thermo Scientific™ R685070) with three Mitsubishi™ AnaeroPack-Anaero Gas Generation Sachets (Thermo Scientific™ R681001). After each cycle, 100 µL of the inoculated media was used to inoculate the next batch. Anaerobic bacterial growth was quantified by measuring absorbance at 600 nm using the Biotek Epoch MicroplateATTORNEY DOCKET NO.222112-2430 Spectrophotometer. After five growth cycles (post 72-hour incubation in anaerobic conditions), no significant microbial growth, as measured by the absorbance at 600 nm, was observed when SLIPs were used as the sole carbon source (FIG.15B). In contrast, the fecal microbial community could grow using porcine mucin as the sole carbon source, demonstrating the community is well-adapted to utilize mucin as a carbon source. This indicates that SLIPs do not provide the conditions suitable for anaerobic growth.

[0401] Transit Along the Gut. SLIPs transport across the gastrointestinal tract over time and their clearance post-treatment were examined (FIG. 16). Fluorescein-tagged SLIPs were tracked in stool samples by first collecting the samples in 200 μL of PBS, then vortexing until dissolved and centrifuging at 5,000 RPM for 5 minutes. The supernatant was then transferred to a black Costar 96-well plate and analyzed using the Cytation™3 instrument (BioTek®) (excitation 487 nm / emission 528 nm). To demonstrate the control and flexibility of the labeling approach, SLIPs were also labeled with Nile Blue instead of Fluorescein. SLIPs tagged with Nile Blue were administered via drinking water at 6.66 mg / ml. Fluorescence readings from experimental samples were normalized by dividing the raw fluorescence units (FU) by their respective control. Wild-type mice (n=3) received SLIPs daily via drinking water (6.66 mg per mL), with fecal pellets sampled up to 8 days (192 hours). Treatment stopped on day 6 (144 hours), and water was provided instead. Fluorescence plateaued 20 hours after starting treatment and remained constant until the treatment was interrupted. Once access to SLIPs was removed, fluorescence decreased exponentially.

[0402] Diffusion of nutrients. It is beneficial for the hydrogel to maintain its protective barrier function while allowing essential nutrient diffusion, which is relevant for maintaining cell viability. It is beneficial for SLIPs to trap bacterial antigens and toxins while also allowing nutrients to pass. The ability of SLIPs to allow sugar to cross the synthetic mucus barrier by glucose diffusion assays was measured. SLIPs were prepared at 40 mg / mL concentration in phosphate-buffered saline (PBS), with pH adjusted to 1.5, 4.0, 5.5, 6.6, and 7.8. A 25 mM glucose solution was prepared in PBS at corresponding pH values. The pH values selected for testing (1.5-7.8) reflect key physiological conditions in the gastrointestinal tract. These pH values range from highly acidic conditions found in the stomach (pH 1.5-4.0) to the near- neutral environment of the small intestine (pH 6.6-7.8).

[0403] The SLIP-glucose mixture was placed in the apical chamber of the transwell system, and samples from the basal chamber were collected after 24-hour incubation at room temperature. Triplicates were performed per experimental conditions along with controls. Glucose concentrations were determined using a colorimetric glucose assay kit (MAK476, Sigma-Aldrich), measuring absorbance at 570 nm. The kit utilizes glucose oxidase and peroxidase enzymes to generate a colored product proportional to glucose concentration. Standard curves were generated using 25 mM glucose solutions prepared at each tested pHATTORNEY DOCKET NO.222112-2430 value. The diffusion ratio was normalized by dividing the glucose concentration measured after 24 hours in samples containing SLIPs by the glucose concentration in control samples without SLIPs at the same pH. A ratio of 1.0 indicates equivalent glucose diffusion between SLIP and control samples, while values below 1.0 indicate reduced diffusion through the SLIP barrier compared to controls. Statistical analysis of glucose diffusion based on pH condition was performed using one-way Analysis of Variance (ANOVA) followed by a post-hoc Tukey test (FIGS.17A-17B).

[0404] At a physiologically neutral pH, SLIPs exhibited the highest diffusion ratio, indicating enhanced glucose transport compared to control conditions (FIG. 17B). Neutral and slightly acidic conditions (pH 7.8, 6.6, and 5.5) maintained relatively high diffusion, suggesting efficient nutrient transport. As the environment became more acidic, glucose diffusion was reduced, indicating restriction of glucose transport through the SLIP barrier under highly acidic conditions. The reduction in glucose diffusion at acidic pH is consistent with physiological processes, as nutrient absorption does not typically occur in the stomach (FIG. 18). This selective permeability aligns with the functional role of the hydrogel, reinforcing its suitability as a barrier that preserves glucose diffusion at neutral or near-neutral pH, corresponding to the conditions of the small intestine. Thus, the observed results do not pose concerns regarding nutrient absorption, as the hydrogel effectively permits glucose transport in the relevant absorptive regions of the gastrointestinal tract.

[0405] SLIPs capacity to trap bacterial Lipopolysaccharides (LPS). SLIPs are designed for mucoadhesion with mucosal membranes and mucointegration with the residual natural mucus; their non-biodegradability prevents microbial degradation and access for bacterial para- or transcellular invasion. The phenylboronic acid functional group (sticky) increases targeted binding to cells, biological mucins, and the sialic acid component of the bacterial LPS by reversible covalent and non-covalent bonding. SLIPs also form dynamic covalent bonds with sialic acid residues of bacterial lipopolysaccharides (LPS), trapping the microbes and facilitating toxin clearance. Without wishing to be bound by theory, it is possible that SLIPs act as a selective trapping barrier, preventing the activation of pro-inflammatory responses to LPS and other bacterial toxins. Since bacteria often mimic host cells by displaying sialic acid on surface structures (e.g., LPS, flagella, capsular polysaccharides), SLIPs can disrupt this immune evasion strategy by binding to these residues, thereby neutralizing LPS activity, aiding antigen clearance, and reducing pathogenic load.

[0406] FITC-Lipopolysaccharides conjugate (cat no. F3665, Sigma-Aldrich) was used to detect and quantify SLIPs-Bacterial LPS binding capacity (illustrated in FIG.19A). SLIPS was mixed with FITC-Lipopolysaccharides conjugate at different concentrations and paramagnetic beads using a 1:1:1 ratio (total volume of 300ul) for 30 minutes and then placed on a magnetic rack. The paramagnetic beads were added to facilitate the separation of bound and unboundATTORNEY DOCKET NO.222112-2430 components in the SLIPs-LPS binding assay. SLIPs were resuspended in sterile water pH 8.6 to ensure that SLIPS, with a pKa of 8.5, predominately exist in a deprotonated state, optimizing binding conditions. In this state, boron becomes negatively charged (forming a tetrahedral boronate ion), which enhances its ability to bind to diols, such as those found in sialic acid residues on bacterial LPS.

[0407] Fluorescence measurements at 528 nm (excitation: 487 nm) quantified unbound FITC- LPS. Experimental controls included sterile water (pH 8.6), FITC-LPS and beads, FITC-LPS alone, SLIPs alone, and paramagnetic beads alone. Fluorescent readings were adjusted by subtracting the fluorescence of background controls. A standard curve of FITC-LPS was generated to quantify the concentration of trapped LPS under SLIPs exposure (FIG. 19B). SLIPs were applied at a fixed final concentration of 8.3^mg / mL, while FITC-LPS was tested across a range of concentrations (1–10^µg / mL). Binding efficiency was quantified by measuring the reduction in unbound fluorescent signal, providing an indirect yet robust metric for LPS sequestration by SLIPs (FIG.20).

[0408] To model the binding behavior, a three-parameter logistic regression was applied using the equation: a / (1 + exp(-(Concentration - x0) / b)), where a represents the maximum fluorescence response (upper asymptote), x₀ is the inflection point corresponding to the concentration at which half-maximal binding occurs. B denotes the slope parameter that determines the steepness of the curve. Model fitting was performed using non-linear least squares (via the nls function in R), yielding a strong fit with a residual standard error of 0.05 and a pseudo-R² of 0.99 (FIG.20).

[0409] The fitted model estimated that the maximum binding capacity of SLIPs was 4.48 ± 0.08 ^g of LPS. The calculations imply that each milligram of SLIPs can trap approximately 0.53 ^g of LPS. These findings support the proposed mechanism by which SLIPs trap LPS through dynamic covalent bonding between phenylboronic acid groups on SLIPs and diol- containing sialic acid residues on LPS. This interaction enables SLIPs to efficiently sequester bacterial endotoxins, underscoring their potential as a protective mucosal barrier to limit host exposure to pro-inflammatory bacterial components. 9. In vivo Studies

[0410] Agr2 Knockout Model. To test the in-vivo applicability of SLIPs as a mucin analog, a mouse model lacking a gut mucus barrier predisposed to colitis was employed. The knockout of anterior gradient 2 (Agr2) by exon 2-3 removal results in the misfolding and secretory inhibition of Muc2, the dominant intestinal mucin. Wildtype (7 weeks) and Agr2 KO mice (8 ± 2 weeks) were randomized into control and treatment groups (n=7-9 per treatment) at a dose of 6.66 mg / ml (approximately 40 mg / mouse / day) for 17 days.

[0411] Mice were housed in a 12-hour light-dark cycle at a controlled temperature (20-25°C) and humidity (31-67%), with free access to a standard diet. Autoclaved water or SLIP-treatedATTORNEY DOCKET NO.222112-2430 water was provided ad libitum. Health and well-being were monitored daily, and mice were euthanized according to animal care guidelines.

[0412] The generation of Agr2 knockout mice has been described previously (Ref.20). Briefly, the heterozygous knockouts purchased from Jackson Laboratory (B6.129S4(FVB)- Agr2<tm1.2Erle> / J, strain: Strain #:025630) were interbred to produce experimental homozygous KO and wild type littermate control mice. Genotyping for Agr2 KO mice was performed by PCR using tail DNA and a set of 20629 (5′- GGT TTG GGC CTG AAA CTC TG -3′), 20630 (5′- ACC ATC AAG GGT CTG TTG CT -3′), and 20631 (5′- GGC CAT GGG TAC CTT TAG TG -3′) primers. The expected PCR product for the mutant is a single band of 400 bp. For heterozygous, it is two bands, one of 252 bp and another of 400 bp. Wild-type mice produce a single band of 252 bp.

[0413] Fecal samples were collected every two days and immediately preserved at -80ºC. Genomic DNA was extracted using the QIAmp Fast DNA Stool Mini Kit (QIAGEN, Germany; Cat No.#51604) following the manufacturer's protocol. DNA was quantified using the dsDNA High Sensitivity Assay (Thermo Fisher Scientific, Massachusetts, USA) on the Qubit 4.0. The 16S rRNA gene was amplified using LongAmp Hot Start Taq 2X Master Mix (New England Biolabs, Inc.) with two universal 24-mer-barcoded-primers: 27F (AGAGTTTGATCMTGGCTCAG-3’) and 1492R (CGGTTACCTTGTTACGACTT-3’). The fusion barcode primer was synthesized using the same sequences as the Native Barcoding Kit 96 V14 (SQK-NBD114.96). A common 24-base barcode was utilized for each primer pair. Library preparation adhered to the Ligation sequencing DNA V14 (SQK-LSK114, Oxford Nanopore Technologies) protocol and the final library was loaded onto an R10.4.1 (FLO- MIN106) flow cell and sequenced using a GridION or MinION device for 72 hours. The raw nanopore signal was basecalled using Dorado v5.0.0 on the super high accuracy model, sup. The contingency table, assembled at the bacterial species level, was obtained using the RESCUE pipeline (Ref.38). The dataset was rarefied to the minimum library size to perform all microbiome analyses as previously recommended (Ref. 42). Differential abundance analysis was performed using ALDEx2.

[0414] The Differential abundance of species in treated KO mice shows increased SCFA producers and decreased opportunistic pathogens (FIG.21). Notably, Roseburia hominis, R. intestinalis, and R. faecis, known for their roles in producing short-chain fatty acids (SCFAs) like butyrate, were significantly increased in SLIP-treated Agr2- / -mice compared to controls. Additionally, Anaerotruncus colihominis, Oscillibacter valericigenes, and Alistipes communis, which also contribute to SCFA production, were significantly elevated, promoting eubiosis and supporting gut barrier integrity. Pathogenic species Klebsiella variicola and Klebsiella pneumoniae showed substantial reductions, which are beneficial as these bacteria are often associated with dysbiosis and IBD exacerbation.ATTORNEY DOCKET NO.222112-2430

[0415] The SLIPs treatment re-established the gut anaerobiosis as indicated by the significant reduction in the relative abundance of fecal anaerobic microbes to levels similar to those found in the wild-type mice (FIGS.22A and 22B).

[0416] Colon samples were obtained for proteome analysis. The quantification was done using nano-liquid chromatography-tandem mass spectrometry (Nano-LC / MS / MS). Gene Set Enrichment Analysis (GSEA) and Ingenuity Pathway Analysis (IPA) software (v.24.0.1, Qi- agen, USA) were utilized to integrate and interpret proteomic data from colon samples of Agr2 KO mice, comparing untreated controls to those treated with 40 mg / day of SLIPs for 17 days.

[0417] FIG. 23A shows significantly reduced enrichment of immune response processes, pathogen defense, and symbiont interactions in SLIP-treated mice. Pro-inflammatory cytokine regulators were inhibited following treatment (FIG. 23B). The mechanism driving immune deactivation promoted by treatment with SLIPs was modeled, which is interlinked with the inhibition of lipopolysaccharide. Based on the proteome analysis, FIG. 23C illustrates the mechanism of immune deactivation, showing that SLIPs inhibit LPS activity. In this pathway, STAT3 serves as a central mediator, integrating signals from pro-inflammatory cytokines like TNF and IL-6 as well as from IFNG through the activation of STAT1. Additionally, LPS initiates this cascade, with predicted activations such as STAT6 and inhibitions such as STAT1 shaping the immune response. Additional influences were exerted by transcription factors such as MYC and TP53, which are key regulators of cell survival and proliferation in inflammation.

[0418] DSS-induced Colitis Model. In the DSS model, the sulfated polysaccharide does not directly induce intestinal inflammation but acts as a direct chemical toxin to colonic epithelium, resulting in epithelial cell injury. This model is widely used and well-described in the literature (Ref. 24) and morphologically and symptomatically resembles epithelial damage seen in human ulcerative colitis. Wild-type C57BL / 6 mice (8 weeks old) were randomized and separated into treatment groups to ensure unbiased distribution, with six mice in each group (3 males and 3 females per group). Mice received 2.5% (w / v) DSS on days 1-4 to induce acute colitis and then were given 40 mg / day of SLIPs via water ad libitum for 4 days. The DSS- control group received DSS but was not treated with SLIPs. Fecal sample collection and body measurements were done every other day until the end of the experiment. The disease activity index (DAI) was calculated based on changes in body weight, stool consistency, and rectum bleeding. The DAI is the combined weight loss score compared to initial weight, stool consistency, and bleeding. Scores are defined as follows: weight loss: 0 (no loss), 1 (1-5%), 2 (5-10%), 3 (10-20%), and 4 (>20%); stool con-sistency: 0 (normal), 2 (loose stool), and 4 (diarrhea); and bleeding: 0 (no blood), 1 (Hemoccult positive), 2 (Hemoccult positive and visual pellet bleeding), and 4 (gross bleeding, blood around anus). The body weight measurements and the Disease Activity index are shown in FIGS.24 and 25.ATTORNEY DOCKET NO.222112-2430

[0419] To assess gut permeability, the fluorescein isothiocyanate conjugated dextran (FITC- Dextran assay 4.4-kDa) was utilized as a marker for gut permeability after inducing colitis for 4 days and SLIPs treatment for 4 days. Only a minimal amount of the 4-kDa molecule can enter the bloodstream in a normal intestinal barrier since it is nondegradable and too large to pass, therefore serving as a marker for a disrupted barrier. The findings revealed that SLIPs treatment led to a 7-fold reduction in gut permeability compared to the untreated group, as illustrated in FIG.26. K. References

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[0468] It will be apparent to those skilled in the art that various modifications and variations can be made in the present disclosure without departing from the scope or spirit of the disclosure. Other aspects of the disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the disclosure disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the disclosure being indicated by the following claims.

Claims

ATTORNEY DOCKET NO.222112-2430 CLAIMS What is claimed:

1. A pharmaceutical composition comprising a therapeutically effective amount of at least one block copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the block copolymer comprises a first block and a second block; wherein the first block is selected from a mucoadhesive polymer block and a backbone polymer block; wherein the second block is selected from a mucoadhesive polymer block and a backbone polymer block, provided that the first block and the second block are not the same; wherein the mucoadhesive polymer block comprises at least one mucoadhesive monomer; wherein the backbone polymer block comprises at least one backbone monomer; wherein the backbone polymer block does not contain a mucoadhesive monomer; wherein the mucoadhesive monomer comprises at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and wherein the backbone monomer comprises at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof.

2. The pharmaceutical composition of claim 1, wherein the mucoadhesive monomer is selected from the group consisting of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, 4- (acrylamido)phenylboronic acid, 3-(acrylamido)phenylboronic acid, 2- (acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 2- vinylphenylboronic acid, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, pyridyl disulfide ethyl acrylate, pyridyl disulfide ethyl acrylamide, pyridyl disulfide alkyl methacrylamide 2- (pyridin-2-yldisulfaneyl)ethyl acrylate, 2-(pyridin-2-yldisulfaneyl)ethyl acrylamide, 2- (pyridin-2-yldisulfaneyl)ethyl methacrylate, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

3. The pharmaceutical composition of claim 1, wherein the mucoadhesive monomer is selected from the group consisting of 4-(acrylamido)phenylboronic acid, 3- (acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4-vinylphenylboronicATTORNEY DOCKET NO.222112-2430 acid, 3-vinylphenylboronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

4. The pharmaceutical composition of claim 1, wherein the mucoadhesive monomer comprises at least one residue of phenylboronic acid or a derivative thereof.

5. The pharmaceutical composition of claim 1, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, poly(ethlylene glycol) acrylate, poly(ethylene glycol) methacrylate, 2-(3-acrylamidopropanamido)phenyl)boronic acid, and N-(2-(3-acetamido- 4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

6. The pharmaceutical composition of claim 1, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, and N-(2-(3-acetamido-4,5-dihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

7. The pharmaceutical composition of claim 1, wherein the backbone monomer is selected from acrylic acid and N,N-dimethylacrylamide.

8. The pharmaceutical composition of claim 1, wherein the backbone monomer comprises at least one residue of acrylamide, acrylic acid, or a derivative thereof.

9. The pharmaceutical composition of claim 0, wherein the mucoadhesive polymer block further comprises at least one backbone monomer.

10. The pharmaceutical composition of claim 0, wherein the block copolymer further comprises a third block; wherein the block copolymer comprises a first block comprising a first mucoadhesive polymer block and a third block comprising a second mucoadhesive polymer block; and wherein the first mucoadhesive polymer block and the second mucoadhesive polymer block are terminal polymer blocks.

11. The pharmaceutical composition of claim 10, wherein the first block comprises the same mucoadhesive monomer as the third block.

12. The pharmaceutical composition of claim 10, wherein the first block comprises a different mucoadhesive monomer from the third block.

13. The pharmaceutical composition of claim 1, wherein the block copolymer has a formula represented by the following structure: (A)x–block–(B)y–block–(A)z;ATTORNEY DOCKET NO.222112-2430 wherein block A is the first block comprising a mucoadhesive polymer block; and wherein block B is the second block comprising a backbone polymer block.

14. The pharmaceutical composition of claim 0, wherein the block copolymer has a formula represented by the following structure: (A1)x–block–(B)y–block–(A2)z; wherein block A1is the first block comprising a first mucoadhesive polymer block; wherein block B is the second block comprising a backbone polymer block; wherein block A2is the third block comprising a second mucoadhesive polymer block; and wherein the first block comprises a different mucoadhesive monomer from the third block.

15. The pharmaceutical composition of claim 14, wherein the first block comprises a greater wt% of mucoadhesive monomers than the third block.

16. The pharmaceutical composition of claim 14, wherein the first block comprises at least one mucoadhesive monomer comprising at least one residue of a boronic acid, an organoboron, or a derivative thereof.

17. The pharmaceutical composition of claim 1, wherein the total molecular weight of the block copolymer ranges from about 10 kDa to about 50 MDa.

18. The pharmaceutical composition of claim 1, wherein the total molecular weight of the block copolymer ranges from about 100 kDa to about 10 MDa.

19. The pharmaceutical composition of claim 1, wherein the total molecular weight of the block copolymer ranges from about 0.5 MDa to about 5 MDa.

20. The pharmaceutical composition of claim 1, wherein the block copolymer is from about 1% to about 90% of the at least one mucoadhesive polymer block by weight.

21. The pharmaceutical composition of claim 1, wherein the block copolymer is from about 1% to about 25% of the at least one mucoadhesive polymer block by weight.

22. The block copolymer of claim 1, wherein the block copolymer is from about 1% to about 10% of the at least one mucoadhesive polymer block by weight.

23. The pharmaceutical composition of claim 1, wherein the mucoadhesive polymer block is comprised of monomer units, wherein from about 1% to about 100% of the monomer units are the mucoadhesive monomer.

24. The pharmaceutical composition of claim 1, wherein the mucoadhesive polymer block is comprised of monomer units, wherein from about 10% to about 90% of the monomer units are the mucoadhesive monomer.

25. The pharmaceutical composition of claim 1, wherein the mucoadhesive polymer block is comprised of monomer units, wherein from about 30% to about 70% of the monomer units are the mucoadhesive monomer.ATTORNEY DOCKET NO.222112-2430 26. The pharmaceutical composition of claim 1, further comprising at least one agent known to treat a gastrointestinal condition.

27. The pharmaceutical composition of claim 26, wherein the gastrointestinal condition is an inflammatory disease.

28. The pharmaceutical composition of claim 27, wherein the inflammatory disease is an inflammatory bowel disease.

29. The pharmaceutical composition of claim 28, wherein the at least one agent known to treat the inflammatory bowel disease is an anti-inflammatory drug, an immune system suppressor, an antibiotic, a biologic, or other agent known to treat the inflammatory bowel disease.

30. The pharmaceutical composition of claim 28, wherein the at least one agent known to treat the inflammatory bowel disease is selected from the group consisting of mesalamine, balsalazide, olsalazine, azathioprine, mercaptopurine, methotrexate, tofacitinib, upadacitinib, ozanimod, infliximab, adalimumab, golimumab, certolizumab, vedolizumab, ustekinumab, risankizumab, ciprofloxacin, metronidazole, or combinations thereof.

31. The pharmaceutical composition of claim 26, wherein the gastrointestinal condition is colorectal cancer.

32. The pharmaceutical composition of claim 26, wherein the gastrointestinal condition is an enteric infection.

33. A pharmaceutical composition comprising a therapeutically effective amount of at least one random copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the random copolymer comprises: at least one mucoadhesive unit, wherein the mucoadhesive unit comprises at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and at least one backbone unit, wherein the backbone unit comprises at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof. wherein the mucoadhesive units are randomly distributed throughout the random copolymer.

34. The pharmaceutical composition of claim 33, wherein the mucoadhesive monomer is selected from the group consisting of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, 4- (acrylamido)phenylboronic acid, 3-(acrylamido)phenylboronic acid, 2- (acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 2-ATTORNEY DOCKET NO.222112-2430 vinylphenylboronic acid, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, pyridyl disulfide ethyl acrylate, pyridyl disulfide ethyl acrylamide, pyridyl disulfide alkyl methacrylamide 2- (pyridin-2-yldisulfaneyl)ethyl acrylate, 2-(pyridin-2-yldisulfaneyl)ethyl acrylamide, 2- (pyridin-2-yldisulfaneyl)ethyl methacrylate, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

35. The pharmaceutical composition of claim 33, wherein the mucoadhesive monomer is selected from the group consisting of 4-(acrylamido)phenylboronic acid, 3- (acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

36. The pharmaceutical composition of claim 33, wherein the mucoadhesive monomer comprises at least one residue of phenylboronic acid or a derivative thereof.

37. The pharmaceutical composition of claim 33, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, poly(ethlylene glycol) acrylate, poly(ethylene glycol) methacrylate, 2-(3-acrylamidopropanamido)phenyl)boronic acid, and N-(2-(3-acetamido- 4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

38. The pharmaceutical composition of claim 33, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, and N-(2-(3-acetamido-4,5-dihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

39. The pharmaceutical composition of claim 33, wherein the backbone monomer is selected from acrylic acid and N,N-dimethylacrylamide.

40. The pharmaceutical composition of claim 33, wherein the backbone monomer comprises at least one residue of acrylamide, acrylic acid, or a derivative thereof.

41. The pharmaceutical composition of claim 33, wherein the mucoadhesive unit further comprises at least one backbone monomer.ATTORNEY DOCKET NO.222112-2430 42. The pharmaceutical composition of claim 33, wherein the random copolymer comprises at least two mucoadhesive units, wherein one of the mucoadhesive units comprises at least one mucoadhesive monomer that the other mucoadhesive unit does not.

43. The pharmaceutical composition of claim 33, wherein the total molecular weight of the random copolymer ranges from about 10 kDa to about 50 MDa.

44. The pharmaceutical composition of claim 33, wherein the total molecular weight of the random copolymer ranges from about 100 kDa to about 10 MDa.

45. The pharmaceutical composition of claim 33, wherein the total molecular weight of the random copolymer ranges from about 0.5 MDa to about 5 MDa.

46. The pharmaceutical composition of claim 33, wherein the random copolymer is comprised of monomer units, wherein from about 0.01% to about 90% of the monomer units are the mucoadhesive unit.

47. The pharmaceutical composition of claim 33, wherein the random copolymer is comprised of monomer units, wherein from about 0.01% to about 60% of the monomer units are the mucoadhesive unit.

48. The pharmaceutical composition of claim 33, wherein the random copolymer is comprised of monomer units, wherein from about 1% to about 25% of the monomer units are the mucoadhesive unit.

49. The pharmaceutical composition of claim 33, wherein the random copolymer is comprised of monomer units, wherein from about 1% to about 5% of the monomer units are the mucoadhesive unit.

50. The pharmaceutical composition of claim 33, further comprising at least one agent known to treat a gastrointestinal condition.

51. The pharmaceutical composition of claim 50, wherein the gastrointestinal condition is an inflammatory disease.

52. The pharmaceutical composition of claim 51, wherein the inflammatory disease is an inflammatory bowel disease.

53. The pharmaceutical composition of claim 52, wherein the at least one agent known to treat the inflammatory bowel disease is an anti-inflammatory drug, an immune system suppressor, an antibiotic, a biologic, or other agent known to treat the inflammatory bowel disease.

54. The pharmaceutical composition of claim 52, wherein the at least one agent known to treat the inflammatory bowel disease is selected from the group consisting of mesalamine, balsalazide, olsalazine, azathioprine, mercaptopurine, methotrexate, tofacitinib, upadacitinib, ozanimod, infliximab, adalimumab, golimumab, certolizumab, vedolizumab, ustekinumab, risankizumab, ciprofloxacin, metronidazole, or combinations thereof.ATTORNEY DOCKET NO.222112-2430 55. The pharmaceutical composition of claim 50, wherein the gastrointestinal condition is colorectal cancer.

56. The pharmaceutical composition of claim 50, wherein the gastrointestinal condition is an enteric infection.

57. A pharmaceutical composition comprising a therapeutically effective amount of at least one gradient copolymer, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof and a pharmaceutically acceptable carrier, wherein the gradient copolymer comprises: at least one mucoadhesive unit, wherein the mucoadhesive unit comprises at least one mucoadhesive monomer comprising at least one residue of boronic acid, carboxylic acid, a thiol group, a disulfide group, a primary amine, a secondary amine, a tertiary amine, benzoic acid, acrylic acid, acrylate, acrylamide, or a derivative thereof; and at least one backbone unit, wherein the backbone unit comprises at least one backbone monomer comprising at least one residue of acrylamide, methacrylamide, poly(ethylene glycol) acrylamide, acrylate, methacrylate, poly(ethylene glycol) acrylate, acrylic acid, methacrylic acid, or a derivative thereof; wherein greater than 50% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

58. The pharmaceutical composition of claim 57, wherein about 75% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

59. The pharmaceutical composition of claim 57, wherein about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

60. The pharmaceutical composition of claim 57, wherein greater than 50% to about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

61. The pharmaceutical composition of claim 57, wherein about 75% to about 99% of the mucoadhesive units are contained within a terminal region of the gradient copolymer.

62. The pharmaceutical composition of claim 57, wherein the mucoadhesive monomer is selected from the group consisting of acrylic acid, methacrylic acid, 4-vinylbenzoic acid, 4- (acrylamido)phenylboronic acid, 3-(acrylamido)phenylboronic acid, 2- (acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 2- vinylphenylboronic acid, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, pyridyl disulfide ethyl acrylate, pyridyl disulfide ethyl acrylamide, pyridyl disulfide alkyl methacrylamide 2- (pyridin-2-yldisulfaneyl)ethyl acrylate, 2-(pyridin-2-yldisulfaneyl)ethyl acrylamide, 2- (pyridin-2-yldisulfaneyl)ethyl methacrylate, 2-(pyridin-2-yldisulfaneyl)ethyl methacrylate, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2-ATTORNEY DOCKET NO.222112-2430 acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

63. The pharmaceutical composition of claim 57, wherein the mucoadhesive monomer is selected from the group consisting of 4-(acrylamido)phenylboronic acid, 3- (acrylamido)phenylboronic acid, 2-(acrylamido)phenylboronic acid, 4-vinylphenylboronic acid, 3-vinylphenylboronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-chlorophenyl)boronic acid, 4-((2-acrylamidoethyl)carbamoyl)-3-fluorophenyl)boronic acid, 4-((2- acrylamidoethyl)carbamoy1)-3-bromophenyl)boronic acid, and 4-((2- acrylamidoethyl)carbamoyl)-3-iodophenyl)boronic acid.

64. The pharmaceutical composition of claim 57, wherein the mucoadhesive monomer comprises at least one residue of phenylboronic acid or a derivative thereof.

65. The pharmaceutical composition of claim 57, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, poly(ethlylene glycol) acrylate, poly(ethylene glycol) methacrylate, 2-(3-acrylamidopropanamido)phenyl)boronic acid, and N-(2-(3-acetamido- 4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

66. The pharmaceutical composition of claim 57, wherein the backbone monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, methacrylamide, N,N-dimethylacrylamide, N,N-dialkylacrylamides, N-alkylacrylamides, N,N-dialkyl methacrylamides, N-alkyl methacrylamides, poly(ethylene glycol) acrylamide, poly(ethylene glycol) methacrylamide, and N-(2-(3-acetamido-4,5-dihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethyl)methacrylamide.

67. The pharmaceutical composition of claim 57, wherein the backbone monomer is selected from acrylic acid and N,N-dimethylacrylamide.

68. The pharmaceutical composition of claim 57, wherein the backbone monomer comprises at least one residue of acrylamide, acrylic acid, or a derivative thereof.

69. The pharmaceutical composition of claim 57, wherein the mucoadhesive unit further comprises at least one backbone monomer.

70. The pharmaceutical composition of claim 57, wherein the gradient copolymer comprises at least two mucoadhesive units, wherein one of the mucoadhesive units comprises at least one mucoadhesive monomer that the other mucoadhesive unit does not.

71. The pharmaceutical composition of claim 57, wherein the total molecular weight of the gradient copolymer ranges from about 10 kDa to about 50 MDa.

72. The pharmaceutical composition of claim 57, wherein the total molecular weight of the gradient copolymer ranges from about 100 kDa to about 10 MDa.ATTORNEY DOCKET NO.222112-2430 73. The pharmaceutical composition of claim 57, wherein the total molecular weight of the gradient copolymer ranges from about 0.5 MDa to about 5 MDa.

74. The pharmaceutical composition of claim 57, wherein the gradient copolymer is comprised of monomer units, wherein from about 0.01% to about 90% of the monomer units are the mucoadhesive unit.

75. The pharmaceutical composition of claim 57, wherein the gradient copolymer is comprised of monomer units, wherein from about 0.01% to about 60% of the monomer units are the mucoadhesive unit.

76. The pharmaceutical composition of claim 57, wherein the gradient copolymer is comprised of monomer units, wherein from about 1% to about 25% of the monomer units are the mucoadhesive unit.

77. The pharmaceutical composition of claim 57, wherein the gradient copolymer is comprised of monomer units, wherein from about 1% to about 5% of the monomer units are the mucoadhesive unit.

78. The pharmaceutical composition of claim 57, further comprising at least one agent known to treat a gastrointestinal condition.

79. The pharmaceutical composition of claim 78, wherein the gastrointestinal condition is an inflammatory disease.

80. The pharmaceutical composition of claim 79, wherein the inflammatory disease is an inflammatory bowel disease.

81. The pharmaceutical composition of claim 80, wherein the at least one agent known to treat the inflammatory bowel disease is an anti-inflammatory drug, an immune system suppressor, an antibiotic, a biologic, or other agent known to treat the inflammatory bowel disease.

82. The pharmaceutical composition of claim 80, wherein the at least one agent known to treat the inflammatory bowel disease is selected from the group consisting of mesalamine, balsalazide, olsalazine, azathioprine, mercaptopurine, methotrexate, tofacitinib, upadacitinib, ozanimod, infliximab, adalimumab, golimumab, certolizumab, vedolizumab, ustekinumab, risankizumab, ciprofloxacin, metronidazole, or combinations thereof.

83. The pharmaceutical composition of claim 78, wherein the gastrointestinal condition is colorectal cancer.

84. The pharmaceutical composition of claim 78, wherein the gastrointestinal condition is an enteric infection.

85. A method for the treatment of a disease or disorder in a subject comprising the step of administering to the subject the pharmaceutical composition of claim 1.

86. The method of claim 85, wherein the disease or disorder is associated with mucus depletion in a mucosal membrane.ATTORNEY DOCKET NO.222112-2430 87. The method of claim 85, wherein the disease or disorder is a gastrointestinal condition.

88. The method of claim 85, wherein the disease or disorder is an inflammatory disease.

89. The method of claim 88, wherein the inflammatory disease is an inflammatory bowel disease.

90. The method of claim 85, wherein the disease or disorder is colorectal cancer.

91. The method of claim 85, wherein the disease or disorder is an enteric infection.

92. The method of claim 85, further comprising the step of administering a therapeutically effective amount of at least one agent known to treat a gastrointestinal condition.

93. The method of claim 92, wherein the pharmaceutical composition and the at least one agent are administered sequentially.

94. The method of claim 92, wherein the pharmaceutical composition and the at least one agent are administered simultaneously.

95. The method of claim 92, wherein the pharmaceutical composition and the at least one agent are co-formulated.

96. The method of claim 92, wherein the pharmaceutical composition and the at least one agent are co-packaged.

97. A method for the treatment of a disease or disorder in a subject comprising the step of administering to the subject the pharmaceutical composition of claim 33.

98. The method of claim 97, wherein the disease or disorder is associated with mucus depletion in a mucosal membrane.

99. The method of claim 97, wherein the disease or disorder is a gastrointestinal condition.

100. The method of claim 97, wherein the disease or disorder is an inflammatory disease.

101. The method of claim 100, wherein the inflammatory disease is an inflammatory bowel disease.

102. The method of claim 97, wherein the disease or disorder is colorectal cancer.

103. The method of claim 97, wherein the disease or disorder is an enteric infection.

104. The method of claim 97, further comprising the step of administering a therapeutically effective amount of at least one agent known to treat a gastrointestinal condition.

105. The method of claim 104, wherein the pharmaceutical composition and the at least one agent are administered sequentially.

106. The method of claim 104, wherein the pharmaceutical composition and the at least one agent are administered simultaneously.

107. The method of claim 104, wherein the pharmaceutical composition and the at least one agent are co-formulated.

108. The method of claim 104, wherein the pharmaceutical composition and the at least one agent are co-packaged.ATTORNEY DOCKET NO.222112-2430 109. A method for the treatment of a disease or disorder in a subject comprising the step of administering to the subject the pharmaceutical composition of claim 57.

110. The method of claim 109, wherein the disease or disorder is associated with mucus depletion in a mucosal membrane.

111. The method of claim 109, wherein the disease or disorder is a gastrointestinal condition.

112. The method of claim 109, wherein the disease or disorder is an inflammatory disease.

113. The method of claim 112, wherein the inflammatory disease is an inflammatory bowel disease.

114. The method of claim 109, wherein the disease or disorder is colorectal cancer.

115. The method of claim 109, wherein the disease or disorder is an enteric infection.

116. The method of claim 109, further comprising the step of administering a therapeutically effective amount of at least one agent known to treat a gastrointestinal condition.

117. The method of claim 116, wherein the pharmaceutical composition and the at least one agent are administered sequentially.

118. The method of claim 116, wherein the pharmaceutical composition and the at least one agent are administered simultaneously.

119. The method of claim 116, wherein the pharmaceutical composition and the at least one agent are co-formulated.

120. The method of claim 116, wherein the pharmaceutical composition and the at least one agent are co-packaged.

121. A kit comprising the pharmaceutical composition of claim 1 and one or more of: a) at least one agent known to treat a gastrointestinal condition; b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane; c) instructions for treating a gastrointestinal condition; or d) instructions for treating a disease or disorder associated with mucus depletion in a mucosal membrane.

122. The kit of claim 121, wherein the pharmaceutical composition and the at least one agent are co-formulated.

123. The kit of claim 121, wherein the pharmaceutical composition and the at least one agent are co-packaged.

124. A kit comprising the pharmaceutical composition of claim 33 and one or more of: a) at least one agent known to treat a gastrointestinal condition; b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane; c) instructions for treating a gastrointestinal condition; orATTORNEY DOCKET NO.222112-2430 d) instructions for treating a disease or disorder associated with mucus depletion in a mucosal membrane.

125. The kit of claim 124, wherein the pharmaceutical composition and the at least one agent are co-formulated.

126. The kit of claim 124, wherein the pharmaceutical composition and the at least one agent are co-packaged.

127. A kit comprising the pharmaceutical composition of claim 57, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more of: a) at least one agent known to treat a gastrointestinal condition; b) at least one agent known to treat a disease or disorder associated with mucus depletion in a mucosal membrane; c) instructions for treating a gastrointestinal condition; or d) instructions for treating a disease or disorder associated with mucus depletion in a mucosal membrane.

128. The kit of claim 127, wherein the pharmaceutical composition and the at least one agent are co-formulated.

129. The kit of claim 127, wherein the pharmaceutical composition and the at least one agent are co-packaged.

130. The use of the pharmaceutical composition of claim 1 in the manufacture of a medicament for the treatment of a disease disorder associated with mucus depletion in a mucosal membrane in a subject.

131. The use of the pharmaceutical composition of claim 33 in the manufacture of a medicament for the treatment of a disease disorder associated with mucus depletion in a mucosal membrane in a subject.

132. The use of the pharmaceutical composition of claim 57 in the manufacture of a medicament for the treatment of a disease disorder associated with mucus depletion in a mucosal membrane in a subject.

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