Benzoxaborole polymers and methods of use

Abstract

To provide benzoxaborole polymers and methods of use.SOLUTION: The disclosure relates to benzoxaborole-containing polymers and to methods of using the same to treat metabolic and gastrointestinal disorders. Suitable cationic polymers that can be modified to include pendant benzoxaborole groups include: vinylic polyamines, polyethyleneimines, acrylamido-based polyamines, chitosan, chemically modified chitosan derivatives such as thiolated chitosan, polyamidoamine (PANAM) dendrimers, poly(vinylpyridine), poly(vinylimidazole), and poly(vinyl aniline).SELECTED DRAWING: None

Description

[Technical field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Application No. 62 / 903,332, filed Sep. 20, 2019, the entire disclosure of which is incorporated herein by reference. [Background technology]

[0002] 2. Background of the Invention Approximately 11.3% of the US adult population suffers from type 2 diabetes mellitus (T2DM), and 35% of US adults have prediabetic conditions. US medical costs attributable to diabetes are approaching $200 billion annually. The incidence of T2DM continues to rise in parallel with the obesity epidemic, and most current treatments for T2DM consist of oral medication regimens that may be suboptimal for many subjects, in part due to side effects associated with systemic absorption of the medication. Bariatric surgery to bypass or remove the duodenum from the digestive tract has been shown to improve T2DM. The Diabetes Surgery Summit recommends bariatric surgery to treat T2DM in some obese patients (grade III obesity) and that bariatric surgery should be considered for the treatment of others (see, e.g., Koliaki, C. et al., BMC Endocr Disord. (2017) 17:50 DOI 10.1186 / s12902-017-0202-6).

[0003] Analysis of the typical diabetic patient path from first-line medication to insulin and surgery and other highly invasive procedures reveals significant gaps, not least ineffective treatments and clinical inertia. Surgery and other solutions have also never been widely adopted. The inclusion of specialty clinicians in the care pathway has contributed to these failures. Thus, effective treatments under the control of a primary care physician are likely to reach a much larger segment of the target population and therefore have a greater impact than those requiring a specialist, e.g., an endocrinologist, gastroenterologist, or surgeon.

[0004] WO 2017 / 024237 discloses certain cationic polymers and the use of the polymers to complex mucus to form an occlusive barrier in the duodenum. Thus, there is a need for new pharmaceuticals and non-invasive methods for treating subjects exhibiting T2MD and associated metabolic disorders. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2017 / 024237 [Non-patent literature]

[0006] [Non-Patent Document 1] Koliaki, C. et al., BMC Endocr Disord. (2017) 17:50 DOI 10.1186 / s12902-017-0202-6 Summary of the Invention [Means for solving the problem]

[0007] Summary of the Invention The present invention provides polymer compositions for forming a physical barrier between the intestinal lining and the luminal contents of the gastrointestinal (GI) tract of a subject. The polymers of the present invention are mucin-interactive agents, which in turn form a physical barrier by interacting with resident mucins in the GI tract. The present inventors have discovered that the incorporation of pendant benzoxaborole groups into certain cationic polymers can dramatically improve the mucin complexing activity of the polymer.

[0008] As shown in the examples, the polymers described herein have improved mucin and mucus complexing activity compared to comparable cationic polymers and can effectively concentrate mucin and mucus at duodenal pH. The polymers bind tightly to mucus at duodenal pH and, once bound to mucus, are resistant to removal by high concentrations of salt (e.g., 1M NaCl). The resulting polymer-mucous complexes have dramatically different properties compared to free mucus. Without wishing to be bound by any particular theory or mechanism, these data demonstrate that the polymers, when administered orally, are capable of complexing mucus in the duodenum to form an obstructive nonabsorbable luminal barrier. The formation of an obstructive barrier in the duodenum is functionally similar or equivalent to bypassing, removing, or resecting the duodenum by surgical or endoscopic procedures, resulting in improved glucose homeostasis (e.g., Koliaki, C. et al., BMC Endocr Disord. (2017) 17:50 DOI 10.1186 / s12902-017-0202-6).

[0009] The present disclosure relates to cationic polymers containing pendant benzoxaborole groups attached directly or indirectly via covalent bonds to the polymer backbone, and methods of treating metabolic diseases comprising administering a therapeutically effective amount of such polymers to a subject in need thereof.

[0010] Another aspect of the present invention is a pharmaceutical composition comprising the polymer of the present invention together with a carrier or diluent.The pharmaceutical composition may be used for therapy, for example, in the treatment of disorders as described herein.Similarly, the present invention provides the use of the polymer disclosed herein as a medicament, and the use of the polymer disclosed herein in the manufacture of a medicament for treating disorders as described herein. [Brief description of the drawings]

[0011] [Figure 1]FIG. 1 illustrates the mucin complexation assay. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Detailed Description of the Invention The present disclosure relates to cationic polymers containing pendant benzoxaborole groups directly or indirectly bonded to the polymer backbone via covalent bonds. The cationic polymer is preferably a polycation that contains amine or ammonium-containing repeat units and may optionally contain other cationic groups, such as imidazolyl, pyridinyl, and guanidino. The cationic polymer preferably contains cationic repeat units and benzoxaborole repeat units. The pendant benzoxaborole groups are substituted with one or more suitable substituents, including electron-withdrawing groups, electron-donating groups, as described herein. In some embodiments, the polymer may contain repeat units that contain cationic groups and benzoxaborole groups. The cationic polymer may also be a copolymer that contains any desired neutral or anionic repeat units, as described further herein, provided that the polymer retains a net cationic charge.

[0013] Also disclosed are pharmaceutical compositions comprising these polymers and methods of treatment using these polymers to treat metabolic disorders, such as type 2 diabetes mellitus (T2DM), type 1 diabetes mellitus (T1DM), prediabetes, hyperlipidemia, obesity, overweight, metabolic syndrome, nonalcoholic steatohepatitis, nonalcoholic fatty liver and polycystic ovary syndrome (PCOS).

[0014] Exemplary polymers include pendant benzoxaborole repeat units of formula (I):

[0015] [ka]

[0016] (In the formula,

[0017] [ka] is a polymer repeat unit;

[0018] Q is a direct bond or a linker;

[0019] X 1 and X 2 are hydrogen, halo, -CN, -NO2, -CF3, -SO2CF3, -SO3(R 1 ), -SO2(R 1 ), -N + (R 1 )(R 2 )(R 3 ), -CON(R 1 )(R 2 ), -COO(R 1 ), substituted or unsubstituted alkyl, -NHCO(R 1 ), -(CH2) m -N(R 1 )(R 2 ), -OCO(R 1 ) and -O(R 1 ) independently selected from the group consisting of;

[0020] R 1 , R 2 and R 3 is, at each occurrence, independently, hydrogen or substituted or unsubstituted alkyl (preferably substituted or unsubstituted C1-C6 alkyl);

[0021] n is an integer from 1 to 100,000;

[0022] and m is an integer from 0 to 4.

[0023] In some embodiments of formula (I), X 1 are halo, -CN, -NO2, -CF3, -SO2CF3, -SO3(R 1 ), -SO2(R 1 ), -N + (R 1 )(R 2)(R 3 ), -CON(R 1 )(R 2 ) or -COO(R 1 ) and X 2 is hydrogen.

[0024] In some embodiments of formula (I), X 1 is a substituted or unsubstituted alkyl, -NHCO(R 1 ), -(CH2) m -N(R 1 )(R 2 ), -OCO(R 1 ) or -O(R 1 ) and X 2 is hydrogen.

[0025] In some embodiments of formula (I), X 1 is halo, preferably fluoro; X 2 is hydrogen.

[0026] In some embodiments of formula (I), X 1 is halo, preferably fluoro; X 2 is halo, preferably fluoro.

[0027] In some embodiments of formula (I), X 1 is -CF3, and X 2 is hydrogen.

[0028] In some embodiments of formula (I), X 1 is hydrogen, and X 2 is hydrogen.

[0029] -F, -Cl, -CN, -NO2, -CF3, -SO2CF3, -SO3(R 1 ), -SO2(R 1 ), -N + (R 1 )(R 2 )(R 3 ), -CON(R 1 )(R 2 ) and -COO(R 1It is advantageous to have substituents such as 1,2-dichlorophenyl) on the benzoxaborole since these groups are electron-withdrawing in nature and can lower the pKa of the benzoxaborole.

[0030] Without wishing to be bound by any particular theory, it is believed that certain substituted benzoxaborole polymers have a lower pKa than unsubstituted benzoxaborole polymers, thereby providing greater sensitivity to elevated pH between the stomach and duodenum, enhancing the mucin complexing activity of the polymer at the desired barrier formation site.

[0031] In some embodiments, the polymer contains a repeat unit of formula (I), where Q is a direct bond and X 1 , X 2 and n is as previously described.

[0032] In other embodiments, the polymer contains recurring units of formula (I) where Q is -N(R 1 )-(CH2) m -, -(CH2) m -N(R 1 )-C(O)-, -C(O)-N(R 1 )-, -C(O)-(CH2) m -, -C(O)-N(R 1 )-(CH2) m -N(R 1 )-, -(CH2) m -, -O-(CH2) m -, -S-(CH2) m - and -OC(O)-(CH2) m -, and X is a linker selected from the group consisting of 1 , X 2 , m and n are as defined above.

[0033] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described, and Q is -N(R 1 )-(CH2) m-, m is 0 or 1, R 1 is preferably hydrogen.

[0034] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described and Q is -NH-.

[0035] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described and Q is -NH-CH2-.

[0036] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described, and Q is -(CH2) m -N(R 1 )-C(O)-, m is 0 or 1, and R 1 is preferably hydrogen.

[0037] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described and Q is -NH-C(O)-.

[0038] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described and Q is -CH2-NH-C(O)-.

[0039] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described, and Q is -C(O)-N(R 1 )- and R 1 is preferably hydrogen.

[0040] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described and Q is -C(O)-NH-.

[0041] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described, and Q is -C(O)-(CH2) m - and m is 0 or The answer is 1.

[0042] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described and Q is -C(O)-CH2-.

[0043] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described, and Q is -C(O)-N(R 1 )-(CH2) m -N(R 1 )-, m is 0 or 1, and R 1 is preferably hydrogen.

[0044] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described and Q is -C(O)-NH-CH2-NH-.

[0045] In some embodiments, the polymer comprises a repeat unit of formula (I), X 1 , X 2 and n is as previously described, and Q is -(CH2) m -, -O-(CH2) m -, -S-(CH2) m - and -OC(O)-(CH2) m and m is an integer from 0 to 4.

[0046] In some embodiments, the polymer repeat units are composed of cationic repeat units, amine-functionalized repeat units and nitrogen-containing repeat units.

[0047] In some aspects, the polymer repeat units are comprised of cationic repeat units.

[0048] In some aspects, the polymer repeat units are comprised of amine-functionalized repeat units.

[0049] In some aspects, the polymer repeat units are composed of nitrogen-containing repeat units.

[0050] Suitable cationic polymers that may be modified to contain pendant benzoxaborole groups include vinyl polyamines, polyethyleneimines, acrylamide-based polyamines, chitosan, chemically modified chitosan derivatives such as thiolated chitosan, polyamidoamine (PANAM) dendrimers, poly(vinylpyridine), poly(vinylimidazole), and poly(vinylaniline).

[0051] Vinyl polyamines that may be modified to contain pendant benzoxaborole groups include poly(allylamine) (PAAn), poly(diallylamine) (PDAAn), and poly(vinylamine).

[0052] Acrylamide-based polyamines which may be modified to contain pendant benzoxaborole groups include poly(acrylamidopropylamine), poly(methacrylamidopropylamine), and related poly(acrylamidoalkylamines).

[0053] Suitable nitrogen-containing repeat units for inclusion in polymers are well known in the art and include, for example, polyvinylamines, poly-N-alkylvinylamines, polyacrylamides, polyalkylacrylamides (e.g., polymethacrylamides), poly-N-alkylacrylamides, polyalkyl-N-alkylacrylamides, polyallylamines, poly-N-alkylallylamines, polydiallylamines, poly-N-alkyldiallylamines, polyethyleneimines, polyaminostyrenes, polyvinylimidazoles, polyvinylpyridines, and the like.

[0054] When the amino nitrogen is protonated, the amine-containing repeat unit is cationic. If desired, the cationic character may be modified using known methods, for example by converting the amine to a guanidino, biguanide, quaternary ammonium, or by introducing additional amino groups, for example by alkylating the amine with an alkylamino or alkylammonium group.

[0055] The cationic polymers are preferably polycations which contain amine or ammonium containing repeat units and may optionally contain other cationic groups such as imidazolyl and pyridinyl.

[0056] Polyamines are typically highly charged at duodenal pH (about pH 5-6), but due to the high density of protonated amine sites, they become slightly deprotonated upon passage from the stomach (pH about 2) to the duodenum (pH about 5) after ingestion. Even a small amount of neutralization effectively reduces the charge density of the polymer, leading to these polymer chains becoming more coiled, compacted, and poorly hydrated as the pH increases. Without wishing to be bound by theory, it is believed that this pH responsiveness contributes to the preferential complexation of mucus in the duodenum compared to the stomach. Other polymers of the invention that may respond to increased duodenal pH contain cationic repeat units (e.g., repeat units with protonated amines) that have derivative or structural features that result in a lower pKa value than that of standard protonated aliphatic amines. The lower pKa of these protonated polymers makes them more sensitive to the increase in pH that coincides with passage from the stomach to the duodenum. As a result, these types of polymers can be targeted to interact with the loose mucus of the proximal duodenum, and include polyamines that are substituted with polar groups, such as hydroxyl groups, less than 3 carbon atoms away from the protonated amine.In some embodiments, the polymer comprises an amine that is modified to have a lower pKa than the unmodified amine.For example, the cationic polymer may have a pKa value of less than 9.0, more preferably less than 8.0, and most preferably less than or equal to 7.0.

[0057] The polymer may be a homopolymer, in which case the polymer may contain nitrogen-containing repeat units (e.g., polyamines or polyamides) with the pendant benzoxaborole groups attached directly or indirectly to the polymer backbone via nitrogen atoms of the repeat units.

[0058] Preferably, the polymer is a copolymer containing repeat units of formula (I) and other repeat units which are preferably cationic (e.g., nitrogen-containing repeat units), but may be neutral or anionic, provided that the polymer carries an overall cationic charge.

[0059] In the polymer disclosed herein, at least about 5% of the repeating chemical units contain pendant benzoxaborole groups.In some cases, substantially all of the repeating chemical units in the polymer contain pendant benzoxaborole groups.Preferably, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 5% to about 20%, or about 5% to about 15% of the repeating chemical units contain pendant benzoxaborole groups.

[0060] Suitable benzoxaborole-containing repeat units for inclusion in the polymers described herein include the following: [ka]

[0061] Suitable benzoxaborole-containing repeat units for inclusion in the polymers described herein include the following: [ka]

[0062] Suitable benzoxaborole-containing repeat units for inclusion in the polymers described herein include the following: [ka]

[0063] Suitable benzoxaborole-containing repeat units for inclusion in the polymers described herein include the following: [ka]

[0064] Suitable benzoxaborole-containing repeat units for inclusion in the polymers described herein include the following: [ka]

[0065] Suitable benzoxaborole-containing repeat units for inclusion in the polymers described herein include the following: [ka]

[0066] Suitable nitrogen-containing cationic monomer repeat units include, but are not limited to, the following: [ka] [ka] [ka] (wherein "n" represents an integer from 1 to 100,000).

[0067] Optionally, the cationic polymer containing pendant boronic acid groups may also contain hydrophobic groups, e.g., pendant hydrophobic groups. As used herein, a hydrophobic group is a moiety that is more soluble (as an individual chemical) in octanol than in water. For example, an octyl substituent is a hydrophobic group because octane is more soluble in octanol than in water. Suitable hydrophobic groups are, for example, C6 or higher linear, branched or cyclic hydrocarbons, which may be substituted with one or more hydroxy, halo, and / or aryl (e.g., benzyl) groups.

[0068] If desired, additional repeat units that may be included in the polymers described herein include neutral and acidic repeat units, such as polyacrylates, polyalkylene glycols, polystyrenes, polyvinyl alcohols, polyvinyl phosphates, polyvinyl sulfates, and the like.

[0069] The copolymers of the present invention may exist in a variety of forms, with suitable forms including block copolymers, graft copolymers, comb copolymers, star copolymers, dendrimers, hyperbranched copolymers, random copolymers, gradient block copolymers, and alternating copolymers.

[0070] The present disclosure also relates to cationic polymers containing pendant hydrophobic groups and the use of such polymers to treat metabolic diseases as disclosed herein.

[0071] Preferably, the polymer disclosed herein is of a size sufficient to ensure that the polymer is not substantially absorbed when orally administered to a subject, such as a human.The molecular weight threshold above which the polymer is not absorbed from the GI tract into the systemic circulation depends on the specific polymer and the conditions in the GI tract as well as other factors, but it is generally recognized that polymers above 1,000 Da are not substantially absorbed from the GI tract into the systemic circulation.Therefore, the composition of the present invention that is not substantially absorbed from the GI tract is substantially free of polymer chains less than 1,000 Da, preferably 5,000 Da, or more preferably 10,000 Da, and has an average molecular weight (M) of at least about 10,000 Da, preferably in the range of 20,000 to 250,000 Da, or more. w The polymers of the invention may contain a distribution of polymer chain lengths and may have a polydispersity index (PDI) in the range of 1.5 to 4.0, but they are substantially free of material less than 1,000 Da, preferably they are free of material less than 5,000 Da, or more preferably they are free of material less than 10,000 Da.

[0072] The polymers of the present invention are soluble and preferably not crosslinked, in some embodiments the polymers may be slightly crosslinked but remain soluble and do not form extensive networks or gels.

[0073] Pharmaceutically acceptable salts of the disclosed polymers are also included in the present invention. For example, polymers with acid functional groups may also be present in anionic form or in combination with a cation in the form of a conjugate base. Suitable cations include alkaline earth metal ions, such as sodium and potassium ions, alkaline earth ions, such as calcium and magnesium ions, and unsubstituted and substituted (primary, secondary, tertiary and quaternary) ammonium ions. Polymers with basic groups, such as amines, may also be protonated and may be combined with a pharma- ceutically acceptable counter anion, such as a halide (Cl). - and Br - ), CH3OSO3 - , HSO4 - , SO4 2- , HCO 3- , CO3 2- , nitrate, hydroxide, persulfate, sulfite, acetate, formate, sulfate, phosphate, lactate, succinate, propionate, oxalate, butyrate, ascorbate, citrate, dihydrogencitrate, tartrate, taurocholate, glycocholate, cholate, hydrogencitrate, maleate, benzoate, folate, amino acid derivatives, nucleotides, lipids, or phospholipids. Similarly, the ammonium group comprises a pharma- ceutically acceptable counterion.

[0074] The polymers disclosed herein are typically provided as a mixture of polymer chains with some variability in chain length. This distribution of polymer chain length can be measured using size exclusion chromatography (SEC) and a detector capable of measuring the molar mass of a polymer, such as multi-angle laser light scattering (MALLS). This method can also confirm the absence of short, low molecular weight polymer chains. The method can also provide a polydispersity index (PDI), which is typically the ratio M w / M n (M w is the weight fraction average molecular weight, and M n is the number average molecular weight). PDI=M w / M n

[0075] M w , M n Values ​​of PDI and PDI can be obtained by SEC, preferably with a MALLS detector. For synthetic polymeric materials made from standard free radical processes, PDI values ​​above 2, or even above 3, are commonly observed. In contrast, living free radical polymerization processes, such as atom transfer radical polymerization (ATR) P) or reversible addition-fragmentation chain transfer (RAFT) can produce materials with PDIs less than 2, or even less than 1.5.

[0076] The polymers disclosed herein can be prepared using any suitable method, such as by direct polymerization of one, two or more monomers or by polymer modification.

[0077] Polymerization may be carried out using techniques known in the art of polymer synthesis (see, for example, Shalaby et al, ed., Water-Soluble Polymers, American Chemical Society, (See, e.g., John Wiley & Sons, Washington, DC, 1991). Some cationic monomers are It is available as a hydrochloride salt and may be polymerized by methods known in the art, for example, via a free radical addition process. In this case, the polymerization mixture includes a free radical initiator. Suitable free radical initiators include azobis(isobutyronitrile), azobis(4-cyanovaleric acid), 2,2'-azobis(2-amidinopropane) dihydrochloride, potassium persulfate, ammonium persulfate, and potassium hydrogen persulfate. Other suitable initiators include ionizing radiation and ultraviolet light. The free radical initiator is preferably present in the reaction mixture in an amount ranging from about 0.01 mole percent to about 5 mole percent relative to the monomer.

[0078] The polymer modification approach uses polyamines, and the copolymer approach uses acrylamide derivatives. The "M" group in reaction schemes 1, 2, 3 and 4 represents a substituted or unsubstituted benzoxaborole group.

[0079] Polyamines can serve as mucus-interacting agents and starting materials for chemical modification with benzoxaborole groups. Exemplary polyamines include polyethyleneimine, hydroxyethylated polyethyleneimine, polyamidoamine (PAMAM) dendrimers, poly(allylamine) (PAAn) and its copolymers, poly(diallylamine) (PDAAn) and its copolymers, poly(vinylamine) and its copolymers, poly(vinyl imidazole) and its copolymers, poly(vinyl pyridine) and its copolymers, poly(vinyl aniline) and its copolymers, amine-containing acrylamide and methacrylamide copolymers, etc. Preferred polyamines include poly(allylamine) (PAAn), poly(diallylamine) (PDAAn), poly(ethyleneimine) (PEI) and poly(methacrylamidopropylamine) (PMAPAn).

[0080] Polyamine derivatives can be obtained from chemical modification of polyamines via amide-forming chemistry using EDC coupling (Scheme 1). [ka]

[0081] 1% wt / vol of the desired polyamine is prepared in deionized water with the pH adjusted to 5.0. Ethanol or other suitable organic is added to the polymer solution at 50% of the initial polymer solution volume. The M-carboxylic acid to be coupled is placed in water at 25% of the initial polymer solution volume to form a solution or slurry. The EDC coupling agent is dissolved in ethanol or other suitable solvent at 25% of the initial polymer solution volume. The EDC solution is then mixed with the M-carboxylic acid solution or slurry. The combined EDC / M-carboxylic acid solution is added dropwise to the polymer solution via pipette or pressure equalizing dropping funnel over approximately 10 minutes. The reaction solution contains about 0.5% wt / vol of polymer, about 62% vol of water and about 38% vol of ethanol or other suitable organic solvent. The reaction is stirred and the pH is maintained at 5.0. The reaction is stirred at room temperature for about 18 hours while the pH is stabilized at 5.0. The polymer is precipitated with an excess (3 times the volume) of acetone.

[0082] Polyamine derivatives can be obtained from the chemical modification of polyamines via the Michael addition reaction, where the polyamine is the nucleophile and the acrylamide is the Michael acceptor (Scheme 2). [ka]

[0083] 1% wt / vol of the desired polyamine was prepared in deionized water and the pH was adjusted to 8.5. The pH may be increased or decreased depending on the level of modification desired. The desired M-acrylamide is dissolved in ethanol or other suitable solvent at 20% of the initial polymer solution volume. The acrylamide solution is then added to the polymer solution to form a reaction mixture containing about 0.83% wt / vol of polymer with about 83% vol water and about 17% vol ethanol or other suitable solvent. The reaction mixture is heated to 70° C. and stirred for 48 hours. The polymer is precipitated with an excess (3 times the volume) of acetone.

[0084] Polyamine derivatives can be obtained from chemical modification of polyamines by hydroxyalkylation using epoxide ring-opening chemistry (Scheme 3). [ka]

[0085] 2% wt / vol of the desired polyamine is prepared in deionized water and the pH adjusted to 6.0. The pH may be increased or decreased depending on the level of modification desired. The desired M-epoxide is dissolved in water / ethanol (25% / 75%) at 100% of the initial polymer solution volume. This solution is then added to the polymer solution to prepare a reaction solution with approximately 1% wt / vol of polymer, approximately 62% vol of water, and approximately 38% vol of ethanol. The reaction mixture was heated at 60° C. for 48 hours. If the epoxide is not in complete solution at 60° C., a small amount of additional ethanol may be added to aid dissolution. The polymer is precipitated with an excess (3 times the volume) of acetone.

[0086] Polyacrylamide derivatives can be obtained, for example, by polymerization of 3-acrylamidopropylamine with the desired M-acrylate or M-acrylamide (Scheme 4). [ka]

[0087] The desired amount of the desired M-acrylate or M-acrylamide monomer is placed in a 30 ml glass vial equipped with magnetic stirring and N2(g) inlet. The desired M-acrylate or M-acrylamide monomer is dissolved in dimethylformamide or other suitable water-miscible organic solvent. The desired amount of cationic, neutral or anionic comonomer is then added. A small amount of water may be required to fully dissolve the charged comonomer in the binary solvent system. An appropriate amount of AIBN initiator is added. The comonomer solution is purged with N2(g) for at least about 15 minutes. The reaction is then heated at 65°C under N2(g). After heating for several hours, the copolymer solution or suspension is precipitated from acetone. The polymer is isolated by precipitation in acetone. It may be necessary for some polymers to add some water and adjust the pH down to facilitate precipitation. Finally, the product may be dissolved in deionized water, frozen in IPA / dry ice, and lyophilized.

[0088] Thus, in some aspects, the invention provides a method for applying a physical barrier between the intestinal lining and the luminal contents of the gastrointestinal (GI) tract of a subject, the method comprising administering a therapeutically effective amount of a polymer described herein to the GI tract of the subject.

[0089] As used herein, the term "physical barrier" or "luminal barrier" refers to a polymer-mucus complex that prevents or reduces chyme contact with the mucosal epithelium underlying the polymer-mucous complex in the intestinal tract. A physical barrier is created when the polymer binds in situ with anionic mucins contained in the mucus lining the intestinal wall. A physical barrier may be substantially complete or partial. A substantially complete physical barrier extends to substantially cover the entire epithelial lining of a target area, e.g., the proximal duodenum. A partial physical barrier extends to cover a portion of a target area, e.g., a portion of the epithelial lining of the duodenum. For example, a partial barrier may cover at least about 1%, 5%, 10%, 20%, 25%, 30%, 40%, 50% or more of the epithelium at the target site.

[0090] A partial physical barrier may be discontinuous and spatially distributed and may have varying degrees of permeability, for example, a physical barrier may be a semi-permeable complex of a polymer and mucus or mucin on the intestinal luminal surface, preferably in the duodenum.

[0091] In embodiments, the physical barrier or a formulation thereof passes through the subject's natural digestive process, hi yet other embodiments, the physical barrier is removable or reversible by ingesting a liquid or solvent.

[0092] The polymers described herein bind tightly to mucus and mucin to form polymer-mucus complexes that are resistant to dissociation (e.g., by high salt and low pH). Thus, once formed, the physical barrier typically exists for a "retention period" or "dwell time" before being removed by the natural action of the digestive system. Typical retention periods may range from about 30 minutes to about 7 days, including times ranging from about 1 hour to about 3 hours, from about 1 hour to about 5 hours, from about 1 hour to about 24 hours, from about 1 to about 3 days, etc.

[0093] The desired residence time may vary depending on the clinical application and may be adjusted based on the amount of polymer administered, as well as the frequency and interval of administration. For example, up to 50% of subjects presenting with T2DM have gastroparesis or delayed gastric emptying and may require a mucoadhesive lining that remains in place for a longer period of time than prediabetic or nondiabetic obese subjects. Blood glucose levels often spike within the first 2 hours of eating, and most often within the first 60 minutes; therefore, in one embodiment, the lining should be adhered for a minimum of 60 minutes. In another embodiment, a longer lasting mucoadhesive lining may be required for prediabetic subjects who may not take medication before every meal and therefore may not be compatible with treatment and would need to change their behavior. In this application, the lining may be adhered for a minimum of 6-8 hours and may be required for up to 24 hours. Residence time will also be influenced by the mucus layer to which the polymer develops the most affinity. For example, a superficial loosely adherent layer will fall off in minutes to hours, whereas an affinity for a deep, tightly adherent layer will result in a longer lasting mucoadhesive coating. Overall, residence times may be tailored to various clinical and technical considerations in the embodiments outlined in this disclosure.

[0094] The polymers of the present invention are capable of forming an occlusive barrier layer in the proximal intestine, particularly the duodenum. Preferably, the occlusive barrier is formed in the proximal duodenum or duodenal bulb, such that the polymer is released from the stomach and is able to completely form a barrier layer immediately upon entering the proximal duodenum.

[0095] The polymer is orally administered in any suitable dosage form. A variety of dosage forms suitable for oral administration are well known in the art and include liquid formulations (e.g., solutions, suspensions, slurries, syrups), gels, ointments, powders, tablets, caplets, capsules, and the like.

[0096] In one example, polymers may be administered in liquid form, and are typically sufficiently stable and soluble in the stomach to allow immediate delivery to the duodenum in an active state, without needing further swelling, solubilization, or equilibration with the surrounding environment.The polymers described herein are typically polyamines, which undergo some degree of deprotonation when moving from the highly acidic stomach (pH about 2) to the duodenum (pH about 5), which targets the complexing activity of the polymer to the duodenum.However, polymers may also form a barrier layer in the stomach.

[0097] In other examples, the polymer is administered in a solid form that can hydrate in the stomach. The solid form may be formulated to dissolve slowly, which can protect the polymer from the acidity of the stomach, but allows the polymer to enter the proximal duodenum in a fully active state. In other examples, the polymer is administered in the form of an enteric-coated tablet, caplet, capsule, or other enteric-coated dosage form to protect the polymer from the acidity of the stomach. In such examples, the enteric coating is formulated to dissolve or degrade as soon as possible after or during passage through the pyloric valve (when the pH increases from about pH 2), allowing for immediate release of the polymer. Such dosage forms may include a superdisintegrant to facilitate immediate release of the polymer at the desired site in the intestine, for example, the proximal duodenum. Suitable superdisintegrating excipients are well known in the art (see, for example, Mohanachandran, PS et al, Superdisintegrants: An Overview, Int. J. Pharma. Sci. Review and Research, (2011) 6:1 pp 105-109). For example, enteric capsules that can target delivery to the duodenum have been described in the literature (see, for example, Reix N. et al. Intl J Pharm (2012) 422:1-2 pp. 338-340).

[0098] The polymers of the present invention can dissolve rapidly in the stomach, duodenum or on other mucosal surfaces after oral administration.

[0099] In some aspects, pharmaceutical formulations of the polymers of the invention can optionally include a calcium salt, such as calcium chloride or calcium citrate. It is believed that the formation of a physical barrier can be facilitated in the presence of calcium salts.

[0100] If desired, the polymer may be administered to the gastrointestinal tract of the subject via an endoscope, a nasogastric tube, an oral feeding tube, or a similar device. The polymer may also be sprayed onto the mucosa at the desired site of action, e.g., spraying may be performed endoscopically.

[0101] For therapeutic purposes, a "therapeutically effective amount" of the polymer is administered. A therapeutically effective amount, as used herein, is an amount sufficient to affect a desired response, including a clinical response, under the conditions of administration. A therapeutically effective amount may be, for example, sufficient to improve glucose homeostasis, reduce insulin resistance, cause weight loss, and / or improve other signs and / or symptoms of T1DM, T2DM or other metabolic disorders, such as hyperlipidemia, nonalcoholic steatohepatitis, nonalcoholic fatty liver, and other conditions, such as obesity and overweight. For example, a therapeutically effective amount may be an amount sufficient to lower blood glucose levels and / or reduce HbA1C.

[0102] The exact amount to be administered will depend on several known factors, including age, weight, sex, the particular condition to be treated and its severity, sensitivity to the drug, and the overall health of the subject. A skilled clinician can determine the appropriate amount to administer based on these and other factors. Typically, 1 to 5 tablets / capsules are administered per dose, each size being 0 or 0E or 00 or 00E or 000. The dose may be administered 1, 2, 3 or 4 times per day. The timing of dosing will be based on the underlying indication. Preferably, the dose is administered at least 5, 10, 15, 30 or 60 minutes before a meal and within 12 hours before a meal for the treatment of metabolic conditions. For other indications, dosing immediately before or with a meal may be preferred.

[0103] As used herein, and as is well understood in the art, "treatment" is an approach to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results may include, but are not limited to, alleviation or improvement of one or more symptoms or conditions, whether detectable or not, reduction in the severity of a disease or affliction, a stabilized (i.e., non-worsening) state of a disease or affliction, prevention of the spread of a disease or affliction, delay or slowing of the progression of a disease or affliction, improvement or alleviation of the state of a disease or affliction, and remission (whether partial or total). "Treatment" may also mean prolonging survival compared to expected survival in the absence of treatment.

[0104] The present disclosure relates to a method of treating metabolic diseases by administering a therapeutically effective amount of the polymers disclosed herein to a subject in need thereof. Metabolic diseases that can be treated using the method include, for example, glucose intolerance, T1DM, T2DM, prediabetes, hyperlipidemia, obesity, overweight, obesity, dyslipidemia, hypertension, hyperglycemia, impaired glucose tolerance, insulin resistance, metabolic syndrome, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD) and polycystic ovary syndrome (PCOS).

[0105] The present disclosure also relates to a method of treating a gastrointestinal disorder by administering a therapeutically effective amount of the polymers disclosed herein to a subject in need thereof. Gastrointestinal disorders that can be treated using the method include, for example, celiac disease, irritable bowel syndrome, inflammatory bowel disease, colitis, Clostridium difficile, endotoxemia, diarrhea, and constipation.

[0106] The method of the present invention is also useful in addressing leaky gut syndrome and related conditions. Leaky gut syndrome is a technical term that describes a condition in which intestinal permeability increases due to alterations / damage to epithelial tight junctions, resulting in impaired epithelial barrier function. This impaired barrier acts as a conduit for intraluminal macromolecules and antigens to pass through the intestinal wall, triggering inflammatory immunological responses that result in a variety of health conditions. For example, leaky gut syndrome is involved in IBS (irritable bowel syndrome). Certain proteins in food may act as antigens that induce an immune response. For example, in celiac disease, preventing gluten from contacting the epithelium may reduce the immunological response. Leaky gut is also involved in other immunological conditions such as inflammatory bowel disease (Crohn's disease, ulcerative colitis). Strengthening the intestinal barrier can reduce the absorption of endotoxins in the gastrointestinal tract. Some of these endotoxins are the result of normal bacterial metabolism / degradation or due to bacterial overgrowth. This is particularly relevant in patients with impaired liver function, for example in those with cirrhosis, in whom endotoxins are not metabolized (detoxified) by the liver, leading to impaired brain function (referred to as hepatic encephalopathy). The method described herein can be used to strengthen intestinal barrier properties, and can treat or reduce the occurrence of hepatic encephalopathy.Uremia is another condition associated with impaired intestinal barrier function, and can be treated or reduced using the method described herein.Similarly, clinical evidence has demonstrated that intestinal permeability is high in patients with advanced chronic kidney disease (CKD), so the method described herein can be used to treat or reduce the occurrence of CKD.

[0107] Therapeutic methods may also provide benefit by reducing clinical biomarkers associated with various disorders, for example, reducing systemic inflammation, oxidative stress and hyperuricemia.

[0108] The disclosed polymers may be administered to a subject in the form of a pharmaceutical composition that includes a pharma- ceutically acceptable carrier, excipient, buffer, or diluent.

[0109] For oral administration, the pharmaceutical composition of the present invention may be presented in dosage form, such as capsules, tablets, caplets, powders, granules, gels, suspensions, liquids or other suitable dosage forms. Capsules may be gelatin, soft gel or solid. Tablet, caplet and capsule formulations may further contain one or more adjuvants, binders, diluents, disintegrants, excipients, fillers or lubricants, each of which is known in the art. Examples of such include carbohydrates, such as lactose or sucrose, anhydrous calcium hydrogen phosphate, corn starch, mannitol, xylitol, cellulose or its derivatives, microcrystalline cellulose, gelatin, stearate, silicon dioxide, talc, sodium starch glycolate, acacia, flavoring agents, preservatives, buffers, disintegrants and coloring agents.

[0110] Orally administered compositions may contain one or more optional agents to provide a pharma- ceutically palatable preparation, such as sweeteners, e.g., fructose, aspartame, or saccharin; flavoring agents, e.g., peppermint, oil of wintergreen, or cherry; coloring agents; and preservatives. Pharmaceutical formulations suitable for oral administration and methods for preparing them are well known in the art. For example, see Remington: The Science and See Practice of Pharmacy, twentieth edition, 2000.

[0111] The pharmaceutical preparations that may be used orally include push-fit capsules made of suitable materials, such as gelatin, and soft sealed capsules made of suitable materials, such as gelatin, and plasticizers, such as glycerol or sorbitol. Push-fit capsules may also contain active ingredients mixed with fillers, such as lactose, binders, such as starches, and / or lubricants, such as talc or magnesium stearate, and stabilizers as needed. In soft capsules, active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers may be added. All preparations for oral administration should be in dosages suitable for such administration. Methods for encapsulating compositions (e.g., in hard gelatin or cyclodextran coatings) are known in the art (Baker, et al., "Controlled Release of Biological Active Agents", John Wiley and Sons, 1986).

[0112] The method of the present invention involves the co-formulation of a polymer composition with probiotics. The probiotic formulation helps build beneficial probiotic bacteria in the intestinal tract. Probiotics have been shown in the art to significantly affect the reduction of blood glucose, HbA1c, insulin levels and insulin resistance in subjects with diabetes. Suitable probiotics include, but are not limited to, Lactobacillus Bifidobacteria, Saccharomyces boulardii, and Bacillus coagulans, Akkermansia muciniphila, Bifidobacterium spp., Escherichia spp. Methods for preparing formulations containing probiotics are well known in the art.

[0113] Optionally, the therapeutic methods described herein may include co-administering the polymer composition with one or more additional therapeutic agents. The therapeutic agents for co-administration in subjects with diabetes may include GLP-1 receptor agonists, DPP-4 inhibitors, SGLT-2 inhibitors, glucosidase inhibitors, insulin, metformin, sulfonylureas and drugs from the thiazolidenedione classes.

[0114] In certain examples, the additional therapeutic agent is one or more agents indicated for the treatment of diabetes (type 1 and / or type 2), prediabetes, hyperglycemia, impaired glucose tolerance, or insulin resistance. Such agents include biguanides (e.g., metformin), sulfonylureas (e.g., rimepiride, gliclazide, gilpizide, glimepiride, tolbutamide, glibenclamide (glyburide), gliquidone, and glyclopyramide), meglitinides (e.g., repaglinide and nateglinide); thiazolidinediones (e.g., pioglitazone and rosiglitazone), alpha-glucosidase inhibitors (e.g., acarbose and miglitol), dipeptidyl peptidase 4 (DPP4) inhibitors (e.g., vildagliptin, sitagliptin, saxagliptin, and linagliptin), GLP-1 analogs (e.g., exenatide, lixiglitin, saxagliptin, and linagliptin), and the like. senatide, dulaglutide, and liraglutide), sodium-glucose cotransporter 2 (SGLT2) inhibitors (e.g., dapagliflozin, ganagliflozin, and empagliflozin), amylin mimetics (e.g., pramlinitide), D2-dopamine agonists (e.g., bromocriptine), bile acid sequestrants (e.g., cholestyramine, colesevelam, colestilan, and colestimide), and insulins (e.g., human insulin, insulin glulisine, insulin lispro, insulin isophane human, insulin zinc suspension mixed bovine, insulin protamine zinc bovine, insulin isophane porcine, insulin isophane human, etc.).

[0115] Combination therapy can provide several advantages over monotherapy. For example, administration of a polymer and an additional therapeutic agent may enhance the effectiveness and / or reduce the amount of the additional therapeutic agent required for a desired effect. Thus, undesirable side effects of the additional therapeutic agent may be reduced or eliminated. Furthermore, the polymer of the present invention and the additional therapeutic agent may provide superior therapy compared to each agent as a monotherapy, and combination therapy may provide additive or synergistic effects.

[0116] The subject to be treated by the method disclosed herein is typically a mammal, preferably a human subject.Suitable subjects include, but are not limited to, primates, such as humans, monkeys, apes, etc.; cattle, such as cattle, bulls, etc.; sheep, such as sheep; goats, such as goats; pigs, such as pigs, boars, etc.; horses, such as horses, donkeys, zebras, etc.; cats, including wild and domestic cats; dogs, including dogs; rabbits, including rabbits, hares, etc.; and rodents, including mice, rats, etc.Preferably, the subject is a human, including, but are not limited to, fetuses, newborns, infants, juveniles, and adults.

[0117] The terms "a," "an," and "the" as used in this application, including the claims, refer to "one or more." Thus, for example, a reference to "a subject" is to be construed as including "one or more" unless the context clearly indicates otherwise (e.g., a reference to a plurality of subjects). ) includes multiple subjects.

[0118] Throughout this specification and the claims, the terms "comprise," "comprises," and "comprising" are used in a non-exclusive sense, unless the context requires otherwise. Similarly, the term "include" and its grammatical variations are intended to be open-ended, such that the recitation of items in a list does not exclude other similar items that may be substituted for or added to the listed items.

[0119] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation. Unless otherwise defined, 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 subject matter described herein belongs.

[0120] The term "about" when referring to a value means ±20% or ±10. Furthermore, when the term "about" is used in connection with one or more numbers or numerical ranges, it should be understood to refer to all such numbers and include all numbers within the range, modifying that range by expanding the upper and lower limits of the numerical values ​​set forth. Recitation of numerical ranges by endpoints includes all numbers contained within that range, for example, whole integers including these fractions (e.g., recitation of 1 to 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, etc.), and any range within that range.

[0121] As used herein, the term "alkyl" refers to a monovalent aliphatic hydrocarbon, typically containing 1 to about 6 carbon atoms. The alkyl group may be a straight chain, a branched chain, a monocyclic or a polycyclic moiety, or a combination thereof. Suitable substituents for the alkyl group include aryl, -OH, halogen (-Br, -Cl, -I and -F), -O(R'), -O-CO-(R'), -CN, -NO2, -COOH, -NH2, -NH(R'), -N(R')2, -COO(R'), -CONH2, -CONH(R'), -CON(R')2, -S(O)R', -S(O)2R', -SH and -S(R'). Each R' is independently an alkyl group or an aryl group. Substituted alkyl groups may have more than one substituent. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornl, and the like.

[0122] As used herein, the term "alkylene" refers to an optionally substituted -(CH) x - where x is an integer between 1 and 5. Preferably, x is between 1 and 3, more preferably x is 1 or 2. Suitable substituents for an alkylene group are the same as those suitable for an alkyl group.

[0123] As used herein, the term "alkoxy" refers to a group of the formula -O-alkyl. Example alkoxy groups include methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), tert-butoxy, and the like.

[0124] As used herein, the term "aryl" refers to a stable aromatic monocyclic ring system having 3 to 7 ring atoms, all of which are carbon and may be substituted or unsubstituted. Aryl substituents include -OH, halogens (-Br, -Cl, -I and -F), -O(R'), -O-CO-(R'), -CN, -NO2, -COOH, -NH2, -NH(R'), -N(R'), -COO(R'), -CONH2, -CON H(R'), -CON(R'), -S(O)R', -S(O)R', -SH and -S(R'). Each R' is independently an alkyl group.

[0125] As used herein, the term "aryloxy" refers to a group of formula -O-aryl, where the aryl group may be optionally substituted.

[0126] As used herein, the term "electron-withdrawing" refers to an atom or group that attracts electron density from adjacent atoms toward itself, usually by resonance or inductive effects. Suitable electron-withdrawing groups include, but are not limited to, halo, -CN, -NO2, -CF3, -SO2CF3, -SO3(R 1 ), -SO2(R 1 ), -N + (R 1 )(R2 )(R 3 ), -CON(R 1 )(R 2 ) and -COO(R 1 ) is available.

[0127] As used herein, the term "electron donating" refers to an atom or group that donates electron density to a neighboring atom, usually by resonance or inductive effects. Suitable electron donating groups include, but are not limited to, alkyl, amino, -NHCO(R 1 ), -(CH2) m -N(R 1 )(R 2 ), -OCO(R 1 ) and -O(R 1 ) is available.

[0128] As used herein, the term "hydroxy" refers to a group of formula --OH.

[0129] As used herein, "halo" or "halogen" refers to F, Cl, Br, or I.

[0130] As used herein, the phrase "optionally substituted" means unsubstituted or substituted. As used herein, the term "substituted" means that a hydrogen atom has been removed and replaced with a substituent. It is understood that substitution at a given atom is limited by the valence of the atom.

[0131] The following examples are provided to further illustrate embodiments of the present invention, but are not intended to limit the scope of the invention. They are typical of those that might be used, but other procedures, methodologies, or techniques known to those skilled in the art may alternatively be used. EXAMPLES

[0132] I. Synthesis: Reagents: Poly(allylamine hydrochloride) was obtained from Nittobo Medical, Japan (PAAn-HCl, catalog number PAA-HCl-3L, 50.3% solution in water) and used as received. Materials were characterized by 1H-NMR, TGA, and size-exclusion chromatography (SEC-MALLS). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride was obtained from Chem-Impex International, Wood Dale, IL (EDC-HCl, catalog number 00050, 99.8%) and used as received. Materials were characterized by 1H-NMR, FT-IR, and melting point. 1-Hydroxybenzotriazole hydrate was obtained from Chem-Impex International, Wood Dale, IL (HOBt, catalog number 24755, 99.8% (odb), 21.3% water) and used as received. Materials were characterized by 1H-NMR, and FT-IR. 4-Carboxyphenylboronic acid was obtained from Chem-Impex International, Wood Dale, IL (CPBA, Catalog No. 28086, 99.7%) and used as received. Materials were characterized by 1H-NMR, FT-IR, and melting point. 1-Hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-6-carboxylic acid was obtained from Combi-Blocks, San Diego, CA (Cat. No. QB-6821, 95%). Materials were characterized by 1H-NMR, and FT-IR. 1-Hydroxyphenylboronic acid was obtained from Chem-Impex International, Wood Dale, IL (CPBA, Catalog No. 28086, 99.7%) and used as received. Materials were characterized by 1H-NMR, FT-IR, and melting point. 6-fluoro-1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-6-carboxylic acid and 6-fluoro-1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-5-carboxylic acid were obtained from Jiangyin PharmaAdvance Inc., Jiangyin, Jiangsu Province, PRChina, and were fully characterized by HPLC, 1H-NMR, and mass spectrometry. Example 1 Synthesis of PAAn modified with 4-carboxyphenylboronic acid: 50.3% PAAn-HCl solution (5.964 g, 32 mmol amine equivalent) was placed in a 250 ml beaker along with a magnetic stir bar and deionized water (90 ml). The resulting clear solution was magnetically stirred and a pH electrode was placed in it. The pH was adjusted to 8.0 by dropwise addition of 1 N NaOH solution with stirring. 4-Carboxyphenylboronic acid, 99.7% (0.907 g, 5 mmol) was added to the reaction mixture and the resulting suspension was stirred. After 20 min of stirring, the pH had dropped to 6.8 and some solids remained suspended in the solution. Additional NaOH solution was added in portions with stirring to completely dissolve the suspension. The pH of the resulting clear solution was 7.5. Solid HOBt-hydrate powder (0.046 g, 0.25 mmol) was then added to the clear reaction solution and it dissolved after 20 min of stirring. Hydrochloric acid (1N) was added to the reaction mixture to lower the pH to 5.4. The reaction mixture remained clear. EDC-HCl (1.152 g, 6 mmol) dissolved in 10 ml of deionized water was then slowly pipetted into the reaction mixture. The final volume of the clear reaction mixture was 120 ml with a pH of 5.6. The reaction mixture was stirred for 18 hours. The reaction mixture was adjusted to pH=2.5 with 1N HCl and then dialyzed using tangential flow filtration using a Pall Minimate™ TFF system supplemented with saline solution (2.5% NaCl, pH=2.5). After removing 5 diafiltration volumes, the retentate solution was adjusted to pH=8.0 with 1N NaOH. Desalting was then performed by TFF supplemented with deionized water until the conductivity of the filtrate was <200 μS / cm. The retentate solution was collected in a lyophilization bottle. The purified product solution was frozen in an IPA / dry ice slurry and lyophilized until dry (3 days). The yield was 1.492 g as a fluffy white solid. 1H-NMR spectrum (DO / DCl) confirmed the expected structure (15 mol% substitution) and the absence of small molecule impurities. Example 2 Synthesis of PAAn modified with 1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-6-carboxylic acid: 50.3% PAAn solution (0.5017 g, 2.70 mmol amine equivalents) was placed in a 30 ml glass vial along with a magnetic stir bar and deionized water (8.5 ml). The resulting clear solution was magnetically stirred and a pH electrode was placed in it. The solution was adjusted to pH = 8.0 by dropwise addition of 1 N NaOH solution with stirring. 1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-6-carboxylic acid powder (0.0874 g, 0.491 mmol) was added to the reaction mixture and the resulting suspension was stirred. After 20 min of stirring, the pH had dropped to 5.9, with some solids remaining suspended in the solution. Additional NaOH solution was added in portions with stirring to completely dissolve the remaining solids. The pH of the resulting clear solution was 7.4. Solid HOBt hydrate powder (0.0084 g, 0.0491 mmol) was then added to the clear reaction solution, which dissolved after 20 min of stirring. Hydrochloric acid (1 N) was added to the reaction mixture to lower the pH to 5.4. The reaction solution remained clear. EDC-HCl (0.1097 g, 0.572 mmol) dissolved in 1.0 ml of deionized water was then slowly poured into the reaction mixture with a pipette. The final volume of the clear reaction mixture was 10 ml and the pH was 5.4. The reaction mixture was stirred for 18 h. The reaction mixture was precipitated in excess acetone. The precipitated solid was dissolved in water, the pH was adjusted to 2.8 with 1 N HCl, and then purified by dialysis using 6k to 8k MWCO cellulose membrane dialysis tubing (Spectrum Laboratories) against 5 gallons of acidified water (pH = 2-3) for 8 h, then against deionized water for 2 days with three additional changes of water. Dialysis bags The solution contained within was collected in a lyophilization bottle. The solution was frozen in IPA / dry ice and placed on a lyophilizer until dry (3 days). The yield was 243 mg as a fluffy white solid. 1H-NMR spectrum (D2O / DCl) confirmed the expected structure (18 mol % substitution) and the absence of small molecule impurities. Example 3 Synthesis of PAAn modified with 1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-5-carboxylic acid: 50.3% PAAn solution (5.004 g, 26.90 mmol amine equivalents) was placed in a 100 ml glass beaker along with a magnetic stir bar and deionized water (40.0 ml). The resulting clear solution was magnetically stirred and a pH electrode was placed in it. The solution was adjusted to pH 8.1 by dropwise addition of 5 N NaOH solution with stirring. 1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-6-carboxylic acid powder (0.839 g, 4.71 mmol) was added to the reaction mixture and the resulting suspension was stirred. After 20 min of stirring, the pH had dropped to 6.3, with some solids remaining suspended in the solution. Additional NaOH solution was added in portions with stirring to completely dissolve the remaining solids. The pH of the resulting clear solution was 8.1. Solid HOBt hydrate powder (0.039 g, 0.22 mmol) was then added to the clear reaction solution, which dissolved after 20 minutes of stirring. Hydrochloric acid (4N) was added to the reaction mixture to lower the pH from 7.9 to 5.8. The reaction solution remained clear. EDC-HCl (0.900 g, 4.69 mmol) dissolved in 1.25 ml of deionized water was then slowly poured into the reaction mixture with a pipette. The final volume of the clear reaction mixture was about 50 ml with a pH of 5.9. The reaction mixture was stirred for 18 hours. The reaction mixture was adjusted from pH 5.3 to pH 2.6 with 4N HCl and then dialyzed against 5 gallons of acidified water (pH=2.5) using 6k to 8k MWCO cellulose membrane dialysis tubing (Spectrum Laboratories) for 18 hours and then against deionized water for 2 days with two additional changes of water. The solution contained within the dialysis bag was collected in a lyophilization bottle. The solution was frozen in IPA / dry ice and placed on a freeze dryer until dry (3 days). The yield was 2.72 g as a fluffy white solid. 1H-NMR spectrum (DO / DCl) confirmed the expected structure (15 mol % substitution) and the absence of small molecule impurities. Example 4 Synthesis of PAAn modified with 6-fluoro-1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-5-carboxylic acid: 50.3% PAAn solution (4.999 g, 26.88 mmol amine equivalent) was placed in a 100 ml glass beaker along with a magnetic stir bar and deionized water (40.0 ml). The resulting clear solution was magnetically stirred and a pH electrode was placed in it. The solution was adjusted to pH 8.1 by dropwise addition of 5 N NaOH solution with stirring. 6-fluoro-1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborole-5-carboxylic acid powder (0.829 g, 4.23 mmol) was added to the reaction mixture and the resulting suspension was stirred. After 20 min of stirring, the pH had dropped to 6.9, with some solids remaining suspended in the solution. Additional NaOH solution was added in portions with stirring to completely dissolve the remaining solids. The pH of the resulting clear solution was 8.1. Solid HOBt hydrate powder (0.038 g, 0.22 mmol) was then added to the clear reaction solution, which dissolved after 20 minutes of stirring. Hydrochloric acid (4N) was added to the reaction mixture to lower the pH from 8.1 to 5.5. The reaction solution remained clear. EDC-HCl (0.9057 g, 4.72 mmol) dissolved in 1.25 ml of deionized water was then slowly poured into the reaction mixture with a pipette. The final volume of the clear reaction mixture was about 50 ml with a pH of 5.6. The reaction mixture was stirred for 18 hours. The reaction mixture was adjusted from pH 5.5 to pH 2.6 with 4N HCl and then dialyzed using 6k to 8k MWCO cellulose membrane dialysis tubing (Spectrum Laboratories) against 5 gallons of acidified water (pH=2.5) for 18 hours and then against deionized water for 2 more days with two changes of water. The dialysis solution contained within the dialysis bag was 0.01 g. The dried solution was collected in a lyophilization bottle. The solution was frozen in IPA / dry ice and placed on the lyophilizer until dry (3 days). The yield was 2.76 g as a fluffy white solid. 1H-NMR spectrum (DO / DCl) confirmed the expected structure (14 mol % substitution) and the absence of small molecule impurities. II. Mucin Mixture Assay:

[0133] The scheme for the mucin mixture assay is shown in FIG.

[0134] When water-soluble mucin glycoproteins are mixed with soluble polymeric complexing agents, a variety of results can be obtained. The solution may remain clear or may become cloudy or even opaque. The physical state of the mixture may remain homogeneous and fluid, or may contain particles or even take on a gel-like appearance. The opacity and physical appearance of the mixture provides a qualitative assessment of the interaction between the complexing agent and the mucin, and more importantly, the physical properties of the complex that is formed.

[0135] We seek polymers that can form a wide range of network structures when mixed with mucin. We have found that when certain polymers are combined in solution with mucin glycoproteins, complexation can be observed by the appearance of a gel-like or opaque mixture with substantial phase separation. Conversely, in the absence of complexation, the solution can remain clear and fluid. In our search for polymers that can strongly complex mucin, we are interested in classifying such mixtures according to a well-defined description of transparency and physical state.

[0136] For example, in some cases, a dispersion may be formed in which the fine particles are not visible. The dispersion may be stable for several days without settling or agglomeration. In other cases, a suspension of small particles may be formed. The suspension may settle over time (hours, days), but is generally stable and can be resuspended by mixing. There is prior literature for using a turbidity metric derived from optical absorbance at a particular visible light wavelength, and the relationship of this metric to assess the degree of complexation in some related experiments involving complexation with polymers.

[0137] However, in some cases, complexation of mucin with certain polymers results in gross precipitation of larger particles, sometimes with irregular or complex morphology. This can be indicative of extensive network formation. In some cases, these precipitated solids have an adhesive nature and adhere to the walls of the vial in which the mixing experiment is carried out. In some cases, complex fiber-like precipitates or gels are formed. Furthermore, many cases of gross precipitation are accompanied by phase separation (syneresis) in which the polymer / mucin complexes are deposited as an adhesive film or gel-like mass, and a separate liquid phase (usually clear or very slightly hazy) is observed. In cases of gross precipitation such as these, the degree of complexation is likely to be very high, and it is useful to classify the formation and appearance of the mucin / polymer complexes in comparison to the related materials.

[0138] The objective of the mucin mixture assay is to determine whether a test polymer forms an insoluble complex when mixed with soluble mucin glycoprotein under a set of standard conditions and to observe the general properties of the resulting mixture. The results of the assay are: 1) the assignment of a transparency descriptor, 2) the assignment of a physical state descriptor, and 3) an explanation and comment, if necessary.

[0139] Isolation of the water-soluble fraction of Sigma (cat. no. 1778) mucin from porcine stomach type 3 (MPS-3): The protocol is as follows: Mix 1.0 g of MPS-3 with 40 ml of MilliQ deionized water in a 50 mL conical tube. The tube is then transferred to a rotating carousel mixer. The suspension was left overnight by attaching it to the centrifuge. The 50 ml conical tube was then centrifuged in a Beckman Avanti centrifuge (5,300 rpm, 60 min). The supernatant was carefully poured into a new 50 ml conical tube without disturbing the solid pellet at the bottom. The centrifugation of the supernatant was then repeated (Beckman Avanti centrifuge, 5,300 rpm, 60 min). The supernatant was again collected in a weighed 50 ml conical tube. The mucin solution was frozen at -80°C for at least 2 hours. The frozen samples were placed in a freeze dryer for at least 3 days. The freeze-dried product was collected, the tubes were weighed, and the % yield was calculated, expecting a recovery of 0.60-0.65 g. The solids were stored in a refrigerator at 2-5°C.

[0140] Preparation of 0.1M MES-Saline Buffer: 21.3 g of (2-(N-morpholino)ethanesulfonic acid monohydrate) (MES, JT Baker Bioreagent, >98%) was added to a 1 liter bottle. 9.0 g of sodium chloride (Fisher Chemical, USP grade) was added to the bottle. A 1 liter graduated cylinder was filled with mQ deionized water (DIW, 18 MΩ). Approximately 750 ml of DIW was added to the bottle, capped, and shaken to dissolve all solids. A magnetic stir bar and pH electrode were added to the bottle and stirring was started. 1N NaOH solution was added in portions to bring the pH of the solution to 6.0 (or other desired pH) and the volume of solution added was recorded. Additional DIW was added as needed to bring the total volume to 1 liter. The bottle was capped and the buffer was stored at 2-5°C.

[0141] Polymer / Mucin Mixture Turbidity and Observation Assay: A 1.0% w / w solution was made of each test polymer using MES-saline buffer adjusted to pH 6.0 (or other desired pH). A 1.0% w / w solution of water-soluble MPS-3 was made using MES-saline buffer adjusted to pH 6.0 (or other desired pH). The mucin solution was slightly hazy. 0.25-0.5 mL of the MPS-3 solution was placed in a 2-dram (4 mL) vial. An equal volume of the test polymer solution was slowly added to the vial. The vial was capped and swirled gently to mix while simultaneously observing any physical changes. The resulting mixture can be read by choosing one of the following descriptors for the clarity of the mixture: clear (only slightly hazy, similar to the initial mucin solution); hazy (hazier than the initial mucin solution, text can be read through it); cloudy (not transparent, cannot be read through it, but light transmits well); opaque (milk-like, not transparent, not much light transmits through it), and not applicable (material shows gross phase separation). The resulting mixture can be read by choosing one of the following descriptors for the physical state of the mixture: clear; dispersion (no particles visible); suspension (very fine particulates visible), precipitate (large irregular particles, may be sticky) and phase separation (film, gel deposited, clear or hazy supernatant fluid may be visible). The mixture in the capped vial was observed for approximately 1 hour to note any changes.

[0142] Mucin Mixing Observation Assay Results: Following the protocol described above, a series of test compounds were tested for their ability to form insoluble complexes with mucin in MES buffer at pH=5.5, 6.0, and 6.5. Polymer solutions were made and mixed gently on a rotisserie at least 1 hour prior to the experiment. Complete dissolution was confirmed for all test polymers before the start of the experiment.

[0143] [Table 1-1] [Table 1-2]

[0144] The results of the mucin mixing assay show that when the parent polymer PAAn-HCl is mixed with mucin, hazy dispersions are obtained at all pH values. This result suggests that in this case the polymer / mucin complex does not form an extensive network structure. When the modified PAAn of Example 1 is mixed with mucin, similar hazy dispersions are observed at pH=5.5 and 6.0. However, at pH=6.5, strong phase separation is observed with a white film depositing on the container walls. At this higher pH value, the polymer of Example 1 can act as a mucin crosslinker to form an extensive network.

[0145] The polymers of Examples 2, 3 and 4 are structurally related to Example 1. However, the boronic acid group is present as a cyclized benzoxaborole functional group. For the benzoxaborole modified polymers such as Examples 2, 3 and 4, strong phase separation behavior is observed at all pH values ​​tested in this experiment. The benzoxaborole functional group represents an activated phenylboronic acid group that can interact more strongly with mucin at lower pH. Due to this stronger interaction, a stronger and more robust extensive network formation is observed at pH=6.0 and pH=5.5.

[0146] Additionally, a much stronger interaction is observed at pH=5.5 in Example 4 compared to Examples 2 and 3. This is due to the presence of electron-withdrawing fluoro substituents which act to further lower the pKa of the boronic acid moiety. While the present invention has been particularly shown and described with reference to certain preferred embodiments, it should be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. The present invention provides, for example, the following items. (Item 1) A polymer comprising pendant benzoxaborole repeat units of formula (I): [ka] During the ceremony, [ka] is a polymer repeat unit; Q is a direct bond or a linker; X 1 and X 2 are hydrogen, halo, -CN, -NO2, -CF3, -SO2CF3, -SO3(R 1 ), -SO2(R 1 ), -N + (R 1 )(R 2 )(R 3 ), -CON(R 1 )(R 2 ), -COO(R 1 ), substituted or unsubstituted alkyl, -NHCO(R 1 ), -(CH2) m -N(R 1 )(R 2 ), -OCO(R 1 ) and -O(R 1 ) independently selected from the group consisting of; R 1 , R 2 and R 3 is, at each occurrence, independently, hydrogen or substituted or unsubstituted alkyl (preferably substituted or unsubstituted C1-C6 alkyl); n is an integer from 1 to 100,000; m is an integer from 0 to 4; polymer. (Item 2) 2. A polymer comprising pendant benzoxaborole repeat units of formula (I) according to item 1, wherein the polymer repeat units are composed of cationic repeat units, amine-functionalized repeat units and nitrogen-containing repeat units. (Item 3) 3. The polymer comprising pendant benzoxaborole repeat units of formula (I) according to item 2, wherein the polymer repeat units are composed of nitrogen-containing repeat units. (Item 4) The Q is -N(R 1 )-(CH2) m -, -(CH2) m -N(R 1 )-C(O)-, -C(O)-N(R 1 )-, -C(O)-(CH2) m -, -C(O)-N(R 1 )-(CH2) m -N(R 1 )-, -(CH2) m -, -O-(CH2) m -, -S-(CH2) m - and -OC(O)-(CH2) m -, wherein the linker is selected from the group consisting of: (Item 5) [ka] [ka] [ka] (wherein "n" represents an integer from 1 to 100,000). A polymer comprising pendant benzoxaborole repeat units of formula (I) according to item 1, comprising repeat units of at least one cationic monomer selected from the group consisting of: (Item 6) [ka] wherein n is an integer from 1 to 100,000. (Item 7) [ka] wherein n is an integer from 1 to 100,000. (Item 8) [ka] wherein n is an integer from 1 to 100,000. (Item 9) [ka] wherein n is an integer from 1 to 100,000. (Item 10) [ka] wherein n is an integer from 1 to 100,000. (Item 11) [ka] wherein n is an integer from 1 to 100,000. (Item 12) A pharmaceutical composition comprising the polymer of any one of the preceding items and a pharma- ceutically acceptable excipient. (Item 13) 13. The pharmaceutical composition according to item 12, wherein the composition is a liquid, tablet, caplet or capsule. (Item 14) 13. The pharmaceutical composition according to item 12, wherein the composition is an enteric coated tablet, caplet or capsule. (Item 15) 15. The pharmaceutical composition according to item 14, wherein the enteric coated tablet, caplet or capsule is for targeted delivery to the duodenum. (Item 16) A method for treating a metabolic disorder in a subject, comprising administering a therapeutically effective amount of the polymer according to any one of items 1 to 11 or the pharmaceutical composition according to any one of items 12 to 15 to a subject in need thereof. (Item 17) 17. The method of claim 16, further comprising administering to the subject in need thereof another therapeutic agent. (Item 18) The method according to item 16 or 17, wherein the metabolic disorder is glucose intolerance, T1DM, T2DM, prediabetes, hyperlipidemia, obesity, overweight, dyslipidemia, hypertension, hyperglycemia, impaired glucose tolerance, insulin resistance, metabolic syndrome, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD) and polycystic ovary syndrome (PCOS). (Item 19) The method of claim 16 or 17, wherein the metabolic disorder is type 2 diabetes mellitus. (Item 20) A method for treating a gastrointestinal disorder in a subject, comprising administering a therapeutically effective amount of a polymer according to any one of items 1 to 11 or a pharmaceutical composition according to any one of items 12 to 15 to a subject in need thereof.

Claims

[Claim 1] The invention described in the specification.

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