Compounds for treatment of diabetes and related conditions

Abstract

Compounds useful in the treatment or prevention of diabetes and related conditions and in the prevention, inhibition or reversal of amylin-amyloid fiber formation, the prevention of Langerhans island (beta) cell death, the prevention or reversal of the conversion of soluble human amylin to insoluble human amylin, the prevention and inhibition or reversal of cytotoxic amylin fiber formation. The present invention relates to compounds of formula (I): (I).

Description

[0001] Invention Field

[0002] This invention relates to compounds that can be used to treat or prevent diabetes and related conditions, and to prevent, inhibit, or reverse the formation of amylin-amyloid fibrils, prevent Langerhans island (β) cell death, prevent or reverse the conversion of soluble human amylin to insoluble human amylin, and prevent and inhibit or reverse the formation of cytotoxic amylin fibrils. The invention also relates to pharmaceutical compositions comprising the compounds of the invention, the use of said compounds for the above-described purposes, and their use in the preparation of medicaments for the above-described purposes. Background of the Invention

[0004] Misfolded protein aggregates known as amyloids have reportedly played a key role in the pathogenesis of a variety of diseases, including rheumatoid arthritis, atherosclerosis, Alzheimer's disease, Parkinson's disease, Huntington's disease, and diabetes.

[0005] Misfolded human amylin (hA) is a major component of the pancreatic amylin-amyloid protein in diabetic patients and is believed to contribute at least partially to the development and progression of type 2 diabetes. Therefore, human amylin represents a potential target for developing drugs that can prevent or delay the progression of type 2 diabetes.

[0006] Several molecules capable of altering hA-misfolding and aggregation are known in the art, including the broad-spectrum antibiotic tetracycline. However, most of these molecules have drawbacks, such as side effects or off-target activity, making them unsuitable for long-term use. There remains a need for compounds capable of preventing, inhibiting, and / or reversing amylin-amyloid fibril formation, and / or preventing Langerhans island β-cell death, and / or preventing and / or reversing the conversion of soluble human amylin to insoluble human amylin, and / or preventing, inhibiting, and / or reversing cytotoxic amylin fibril formation.

[0007] The object of this invention is to provide compounds that can overcome or at least partially improve some of the above-mentioned disadvantages and / or at least provide the public with a useful alternative.

[0008] Other objects of the invention may become apparent from the following description, which is given by way of example only.

[0009] In this specification, references to external sources of information (including patent specifications and other documents) are generally used to provide context for discussing the features of the invention. Unless otherwise stated, references to such sources in any jurisdiction should not be construed as an admission that the information is prior art or constitutes part of common general knowledge in the art.

[0010] Invention Summary

[0011] In one aspect, the present invention provides a compound of formula (I):

[0012]

[0013] in:

[0014] X is a C1-C5 alkyl, cyclopropyl, or C2-C4 dialkyl ether;

[0015] A is a C1-C6 alkyl group that is absent or optionally substituted with a halogen, a C1-C3 alkoxy or a C1-C3 haloalkoxy, or A is a phenyl group;

[0016] B is either absent or a C1-C6 alkyl group;

[0017] R1 is independently selected from OH, halogen, C1-C3 alkoxy, C1-C3 deuterated alkoxy, C1-C3 haloalkoxy, C1-C3 deuterated haloalkoxy, CO2R3, OCO2R4 and O-glycosyl;

[0018] R3 is H or a C1-C3 alkyl group;

[0019] R4 is a C1-C3 alkyl group; and

[0020] u is 1, 2, or 3;

[0021] R2 is independently selected from OH, halogen, C1-C3 alkoxy, C1-C3 deuterated alkoxy, C1-C3 haloalkoxy, and C1-C3 deuterated haloalkoxy; and

[0022] v is 1, 2, or 3;

[0023] Or its pharmaceutically acceptable salt.

[0024] In some embodiments of the invention, X is CH2. In other embodiments, X is (CH2)2, (CH2)3, or (CH2)4. In other embodiments, X is CH2-O-CH2. In other embodiments, X is cyclopropyl.

[0025] In some embodiments of the present invention, A is a C1-C5 alkyl group substituted with OCH3, OCD3, OCH2F or F.

[0026] In some embodiments of the present invention, R1 is OH, F, CO2H, OCH3, OCD3, OCF2H, OCOEt, OCO2Pr, or O-glycosyl.

[0027] In some embodiments of the present invention, R2 is OH, F, OCH3 or OCD3.

[0028] In some embodiments of the present invention, u is 2 and R1 are each OH, O-glycosyl or OCO2Pr.

[0029] In some embodiments of the present invention, u is 2 and one R1 is OH and the other R1 is CO2H.

[0030] In some embodiments of the present invention, u is 2 and one R1 is OH and the other R1 is OCF2H.

[0031] In some embodiments of the present invention, u is 2 and one R1 is OH and the other R1 is F.

[0032] In some embodiments of the present invention, v is 1 and R2 is OCH3 or OCD3.

[0033] In some embodiments of the present invention, v is 2 and one R2 is OCH3 or OCD3 and the other R2 is F.

[0034] The compounds of the present invention include, but are not limited to, the following:

[0035]

[0036]

[0037]

[0038]

[0039]

[0040]

[0041]

[0042] In another respect, the present invention relates to pharmaceutical compositions comprising a compound of formula (I) and a pharmaceutically acceptable diluent, carrier or excipient.

[0043] In another aspect, the present invention relates to a method for treating or preventing diabetes, amyloid-associated diseases or Langerhans island β-cell death, the method comprising administering to a patient in need an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0044] In another aspect, the present invention relates to a method for inhibiting, preventing or reversing amylin amyloidosis or the formation of one or more amylin-amyloid fibrils or amylin-amyloid plaques in patients who require it, the method comprising administering to the patient who requires it an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0045] In another aspect, the present invention relates to a method for inhibiting, preventing or reversing the formation of amylin amyloidosis or one or more amylin-amyloid fibrils or amylin-amyloid plaques, the method comprising contacting amylin amyloidosis or one or more amylin-amyloid fibrils or amylin-amyloid plaques with an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0046] In one embodiment, the method is a means of inhibiting, preventing, or reversing the formation of pancreatic amylin-amyloid fibrils.

[0047] On the other hand, the present invention relates to a method for treating or preventing diabetes in patients who require it, the method comprising administering to the patients who require it an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0048] On the other hand, the present invention relates to a method for treating or preventing Langerhans island β-cell death in patients who require it, the method comprising administering to the individual who requires it an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0049] In another aspect, the present invention relates to a method for treating or preventing the death of Langerhans island β cells, the method comprising contacting one or more Langerhans island β cells with an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0050] In another aspect, the present invention provides pharmaceutical compositions comprising one or more compounds of formula (I) and optionally comprising a pharmaceutically acceptable carrier or excipient.

[0051] The present invention also relates to compounds of formula (I) used for one or more of the following: treatment or prevention of amylin-amyloid-related diseases, inhibition, prevention or reversal of amylin-amyloid degeneration or amylin-amyloid fibrils or amylin-amyloid plaque formation, inhibition, prevention or reversal of islet amylin-amyloid fibrils, treatment or prevention of diabetes, or treatment or prevention of Langerhans islet β-cell death, for example in patients in need of it.

[0052] The present invention further relates to the use of compounds of formula (I) in the preparation of medicaments for one or more of the following purposes: treating or preventing amylin-amyloid-related diseases, inhibiting, preventing or reversing amylin-amyloid degeneration or amylin-amyloid fibrils or amylin-amyloid plaque formation, inhibiting, preventing or reversing islet amylin-amyloid fibrils formation, treating or preventing diabetes, or treating or preventing Langerhans island β-cell death, for example in patients in need of it.

[0053] The implementation scheme described herein may involve any of the above aspects.

[0054] Other aspects of the invention are not limited to the information in the summary of the invention, but will become apparent from the following description given by way of example only and with reference to the accompanying drawings.

[0055] The present invention can also be broadly defined as consisting alone or together of any part, element, and feature mentioned or indicated in this application specification, as well as any or all combinations of any two or more of said part, element, or feature; when a particular integer mentioned herein has a known equivalent in the field to which the present invention pertains, the known equivalent is considered to be included herein as if listed separately. Brief description of the attached diagram

[0057] Figure 1 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM ZBBP92 ( ) or HC8768 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are the average + / - SEM (n = 3).

[0058] Figure 2 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM ZBBP92 ( ), HC87610 ( ), HC8762 ( ) or HC8768 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are average values ​​for + / - SEM (n = 3).

[0059] Figure 3 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM ZBBP92 ( ), HC8764 ( ), HC8765 ( ) or HC8766 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are average values ​​for + / - SEM (n = 3).

[0060] Figure 4 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM of either ZBBP92 ( ), HC87610 ( ) or HC8769 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are average values ​​for + / - SEM (n = 3).

[0061] Figure 5 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM ZBBP92 ( ) or HC8761 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are the average + / - SEM (n = 3).

[0062] Figure 6 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM ZBBP92 ( ) or HC9534 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are the average + / - SEM (n = 3).

[0063] Figure 7 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 2.5 μM ZBBP92 ( ) or HC9534 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are the average + / - SEM (n = 3).

[0064] Figure 8 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM ZBBP92 ( ), HC9534 ( ), HC9532 ( ) or HC9533 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are average values ​​for + / - SEM (n = 3).

[0065] Figure 9 The figure shows hA (25 μM) and thiosulfate-T (10 μM) in the absence of ( ) or 25 μM ZBBP92 ( ), HC9534 ( ) or HC87682 ( The fluorescence of thiosulfate-T after incubation was determined under the following conditions. The results are average values ​​for + / - SEM (n = 3).

[0066] Figure 10 The results of cell death analysis of CN cells are shown. Values ​​are the mean ± SE of four independent experiments, with each experiment performed in duplicate. P <0.001 compared to the control; # P <0.05; ## P <0.01; ### P <0.001 compared to hA-treated cells.

[0067] Figure 11 The results of cell death analysis of RINm5F cells are shown. Values ​​are the mean ± SE of four independent experiments, with each experiment performed in duplicate. P <0.001 compared to the control; # P <0.05; ## P <0.01; ### P <0.001 compared to hA-treated cells.

[0068] Detailed Explanation

[0069] definition

[0070] The terms “amyloid protein,” “amyloid fibrils,” and “amyloid plaques” refer to protein aggregates that are produced by misfolding of proteins to form forms that promote and / or enhance aggregation and / or can lead to cytotoxicity.

[0071] The term "amyloidosis" refers to the deposition of amyloid protein in the body.

[0072] The terms “amylin amyloid protein,” “amylin-amyloid filaments,” “amylin aggregates,” or “amylin amyloid plaques” refer to amylinoid protein containing human amylin as an insoluble protein component.

[0073] The terms “island amylin amyloid protein,” “island amylin-amyloid filaments,” “island aggregates,” or “island amylin amyloid plaques” refer to amylin amyloid protein containing human amylin as an insoluble protein component, typically found on Langerhans Isles.

[0074] The term "disruption" used in this article refers to both prior disruption and posterior disruption of amylin amyloidosis.

[0075] The term "amylin monomer" refers to a 37-residue peptide hormone produced and secreted by Langerhans Island β cells. The amino acid sequence of the human precursor protein (also known as the islet amylin polypeptide precursor protein) is found in RefSeq NP_000406.1 and UniProtKB / SwissProt A0A024RAU1.

[0076] The term "oligomer" refers to a molecule having two or more monomer units. Amylin oligomers may contain from about 2, 3, 4, 5, 6, 7, 8, 10, 15, 16, 20 or 25 monomer units to about 100 monomer units, or from about 100 to about 1,000, 2,000, 3,000 or more monomer units.

[0077] The term "amylin oligomer" refers to oligomers in which the monomer unit is human amylin.

[0078] In the context of protein aggregation, the term "interference" refers to disrupting the aggregation process through the compounds described herein, thereby slowing down or preventing aggregation.

[0079] The terms “aggregate,” “aggregate,” or “aggregate” refer to the accumulation of proteins (such as human amylin or its precursor proIAPP), particularly in an insoluble form.

[0080] The term "amyloid-associated disease" refers to diseases including, but not limited to, diabetes (including type 2 diabetes (T2D)), metabolic syndrome, syndrome X, glycemic disorders, and insulin resistance.

[0081] The terms “prevention” or “prevention” refer to stopping processes that have not yet begun, such as amylin-amyloid fibrillation, Langerhans island β-cell death, conversion of soluble human amylin to insoluble human amylin, and formation of cytotoxic oligomers.

[0082] The term “inhibition” is used similarly to “prevention,” but refers to stopping a process that has already begun, such as amylin-amyloid fibrillation, Langerhans island β-cell death, the conversion of soluble human amylin to insoluble human amylin, and the formation of cytotoxic oligomers.

[0083] The term “treatment” refers to the suppression or prevention of the development of amyloid-related diseases and / or the reduction, relief or resolution of amyloid-related diseases or one or more of their side effects.

[0084] The term “treatment” refers to the suppression or prevention of the development of amyloid-related diseases and / or the reduction, relief or resolution of amyloid-related diseases or one or more of their side effects.

[0085] The term "reversal" refers to a return of amyloid-related disease to a previous or milder state. The terms "remission" and "regression" are interpreted in a similar way.

[0086] The term "reversal" refers to a return of amyloid-related disease to a previous or milder state. The terms "remission" and "regression" are interpreted in a similar way.

[0087] The term "administration" refers to providing a patient with a therapeutically effective amount of a compound using one or more methods of administration known in the art. These methods include administration via oral, sublingual, intravenous, subcutaneous, transdermal, intramuscular, intradermal, intrathecal, epidural, intraocular, intracranial, inhalation, rectal, vaginal, and other methods.

[0088] The term "therapeutic effective amount" refers to a dose of compound that is sufficient to provide a high enough concentration to achieve the desired results. For example, in some embodiments, a therapeutic effective amount refers to a dose of the compound described herein that is sufficient to produce one or more of the following: prevention, inhibition, or reversal of amylin-amyloid fibrillation, Langerhans island β-cell death, conversion of soluble human amylin to insoluble human amylin, and formation of cytotoxic oligomers.

[0089] The term "pharmaceutically acceptable salt" refers to a salt of a given compound that is suitable for administration as a drug. For example, such salts can be formed by reacting an acid or base with an amine or carboxylic acid group, respectively. Acid / base addition salts are more soluble in aqueous solvents than their corresponding free acid / base forms.

[0090] The term "prodrug" is used in the broadest sense to encompass those derivatives that are converted in vivo into the compounds of the present invention. Such derivatives are readily apparent to those skilled in the art, including compounds in which the free hydroxyl group is converted into an ester derivative or the cyclic nitrogen atom is converted into an N-oxide. Examples of ester derivatives include alkyl esters (e.g., acetates, lactates, and glutamine), phosphate esters, and those formed from amino acids (e.g., valine).

[0091] The term "solvent" refers to a stoichiometric complex formed by a solute (in this invention, the compound of this invention) and a solvent.

[0092] The term "composition" is intended to refer to a formulation comprising an active ingredient carried by an encapsulating material, thereby forming a capsule in which the active ingredient (with or without another carrier) is surrounded by the carrier.

[0093] The terms “including” and “contains” are used in this document in an open-ended, non-restrictive sense.

[0094] The terms “(C1-C6)” and so on refer to portions having 1 to 6 carbon atoms, respectively. In compound terms such as “hydroxyl-(C0-C4)-alkyl”, the option “(C0)-alkyl” refers to a valence bond (i.e., in this case, a directly bonded hydroxyl group), or in the unsubstituted case, “(C0)-alkyl” refers to hydrogen.

[0095] The term "alkyl" refers to a saturated monovalent hydrocarbon group.

[0096] The term "alkenyl" refers to a monovalent hydrocarbon group containing at least one carbon-carbon double bond, where each double bond may have an E or Z configuration.

[0097] The term "alkynyl" refers to a monovalent hydrocarbon group containing at least one carbon-carbon triple bond.

[0098] Alkyl, alkenyl, and alkynyl groups can be linear (i.e., straight-chain) or branched. This also applies when they are part of other groups (e.g., alkyloxy (= alkoxy, O-alkyl), alkoxycarbonyl, or alkyl-substituted amino groups) or when they are substituted. The number of carbon atoms in an alkyl group can be 1–22, for example 1–12, such as 1, 2, 3, 4, 5, or 6, or 1, 2, 3, or 4. Examples of alkyl groups include methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, sec-butyl, isobutyl, and tert-butyl), pentyl (including n-pentyl, 1-methylbutyl, isopentyl, neopentyl, and tert-pentyl), and hexyl (including n-hexyl, 3,3-dimethylbutyl, and isohexyl). The double and triple bonds in alkenyl and alkynyl groups can be in any position. Examples of alkenyl and alkynyl groups include vinyl, propenyl, propenyl (=allyl), butenyl, 2-methylpropenyl, 3-methylbutenyl, hexenyl, hexenyl, propenyl (=propynyl), butenyl, butenyl, hexenyl, or hexenyl-5-alkynyl. Substituted alkyl, alkenyl, and alkynyl groups can be substituted at any position, provided that the corresponding compound is sufficiently stable and suitable for its intended use, such as as a therapeutic substance. The prerequisite that specific groups and compounds of formula (I) are sufficiently stable and suitable for their intended purpose, such as use as a therapeutic substance, generally also applies to the definition of all groups in compounds of formula (I).

[0099] Unless otherwise stated, the term "alkoxy" (also known as "alkyloxy") refers to the group -OR a , where R a It is an alkyl group as defined above, such as methoxy, ethoxy, propoxy, butoxy, etc.

[0100] The term "deuterated alkoxy" refers to an alkoxy group in which one or more hydrogen atoms are replaced by deuterium atoms. Examples include monodeuterated alkoxy, dideuterated alkoxy, and trideuterated alkoxy, such as OCDH2, OCD2H, and OCD3.

[0101] Unless otherwise stated, the term "halogen" refers to fluorine, chlorine, bromine, or iodine, preferably fluorine and chlorine. In some embodiments, the halogen is F. It is understood that in some cases, the fluorine atom can function as an isostere of hydrogen, and therefore, those skilled in the art can substitute one or more hydrogen atoms from alkyl, alkenyl, aryl, and / or cycloalkyl groups for the fluorine atom in some embodiments.

[0102] Unless otherwise stated, the term "haloalkyl" refers to an alkyl group substituted with one or more, preferably one, two or three identical or different halogen atoms, such as -CH2Cl, -CF3, -CH2CF3, -CH2CCl3, etc.

[0103] Unless otherwise stated, the term "haloalkoxy" refers to -OR b Group, wherein R b It is a haloalkyl group as defined above, such as trifluoromethoxy, trichloroethoxy, 2,2-dichloropropoxy, etc.

[0104] Unless otherwise stated, the term "deuterated haloalkoxy" refers to a haloalkoxy group in which one or more hydrogen atoms are replaced by deuterium atoms. Examples include monodeuterated haloalkoxy, dideuterated haloalkoxy, and trideuterated haloalkoxy, such as OCDF2, OCD2F, and OCD3.

[0105] The term “and / or” means “and” or “or”, or both.

[0106] The numerical ranges disclosed herein (e.g., 1 to 10) are intended to include all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10) and any range of rational numbers within that range (e.g., 2 to 8, 1.5 to 5.5, and 3.1 to 4.7); therefore, all subranges of all ranges explicitly disclosed herein are also explicitly disclosed herein. These are merely examples of specific intentions, and all combinations of numerical values ​​between the listed minimum and maximum values ​​are considered to have been expressly stated in a similar manner in this application.

[0107] As used in this specification, the term "comprising" means "consisting of at least part of...". When interpreting expressions in this specification that include this term, the feature beginning with that term in each statement or claim must be present, but other features may also be present. Related terms such as "comprising" should also be interpreted in the same manner.

[0108] The compounds of the present invention

[0109] This invention provides various compounds suitable for treating amylin amyloidosis, including, for example, diabetes. These compounds possess favorable physicochemical and / or therapeutic properties, making them particularly suitable for treating amylin amyloid-related diseases such as diabetes.

[0110] This invention provides a compound of formula (I):

[0111]

[0112] (I)

[0113] in:

[0114] X is a C1-C5 alkyl, cyclopropyl, or C2-C4 dialkyl ether;

[0115] A is a C1-C6 alkyl group that is absent or optionally substituted with a halogen, a C1-C3 alkoxy or a C1-C3 haloalkoxy, or A is a phenyl group;

[0116] B is either absent or a C1-C6 alkyl group;

[0117] R1 is independently selected from OH, halogen, C1-C3 alkoxy, C1-C3 deuterated alkoxy, C1-C3 haloalkoxy, C1-C3 deuterated haloalkoxy, CO2R3, OCO2R4 and O-glycosyl;

[0118] R3 is H or a C1-C3 alkyl group;

[0119] R4 is a C1-C3 alkyl group; and

[0120] u is 1, 2, or 3;

[0121] R2 is independently selected from OH, halogen, C1-C3 alkoxy, C1-C3 deuterated alkoxy, C1-C3 haloalkoxy, and C1-C3 deuterated haloalkoxy; and

[0122] v is 1, 2, or 3;

[0123] Or its pharmaceutically acceptable salt.

[0124] In some embodiments of the invention, the compound incorporates one or more deuterium atoms in place of hydrogen atoms. Depending on the compound's metabolic pathway and the location of the deuterium, deuteration can be metabolically quiescent, thus enabling its use as a PK tracer; or it can alter the compound's metabolism, thereby acting as a mechanism probe. Introducing deuterium into pharmacologically active substances offers potential benefits, such as improved exposure and reduced production of toxic metabolites, thereby improving efficacy, tolerability, or safety. Deuterated drugs can exhibit different PK characteristics compared to their "hydrogen-only" equivalents.

[0125] The compounds of this invention may exist in crystalline form or as solvates (e.g., hydrates), both of which are intended to be included within the scope of this invention. The solvent should not interfere with the biological activity of the solute. Solvents may be, for example, water, ethanol, or acetic acid. Solventization methods are generally known in the art.

[0126] Unless otherwise stated, the structures shown herein are also intended to include all stereochemical forms of the structure, i.e., the R and S configurations of each asymmetric center. Therefore, single stereochemical isomers of this compound, as well as enantiomers and diastereomeric mixtures, are included within the scope of this invention. Consequently, this invention covers single diastereomers or enantiomers substantially free of other isomers (on a molar basis, >90%, preferably >95%, free of other stereoisomers), and mixtures of such isomers.

[0127] Specific optical isomers can be obtained by resolving racemic mixtures using conventional methods, such as by forming diastereomer salts or treating with optically active acids or bases. Examples of suitable acids include tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, benzoyltartaric acid, and camphorsulfonic acid. The diastereomer mixture is then separated by crystallization, subsequently releasing the optically active base from these salts. A different method for separating optical isomers involves using a chiral chromatographic column optimized to maximize enantiomer separation. Another method involves synthesizing covalent diastereomer molecules by reacting the compounds of the present invention with an optically pure acid or an optically pure isocyanate in its active form. The synthesized diastereomers can be separated by conventional methods (e.g., chromatography, distillation, crystallization, or sublimation) followed by hydrolysis to yield the enantiomerically pure compound.

[0128] The optically active compounds of the present invention can be obtained by using active starting materials. These isomers can be in the form of free acids, free bases, esters, or salts.

[0129] When the compounds of this invention require purification, chromatographic techniques such as high-performance liquid chromatography (HPLC) and reversed-phase HPLC can be used. The compounds can be characterized by mass spectrometry and / or other suitable methods.

[0130] When a compound contains one or more protonable or deprotonable functional groups (e.g., under physiological pH conditions), the compound can be prepared and / or isolated as a pharmaceutically acceptable salt. It is understood that the compound may be in zwitterionic form under given pH conditions.

[0131] In some embodiments, a pharmaceutically acceptable salt of one or more of the compounds described herein is administered to a patient in a therapeutically effective amount. Such salts can be prepared by methods known in the art involving reacting the compound with a suitable organic or inorganic acid or base. Representative organic salts include methanesulfonates, acetates, oxalates, adipates, alginates, aspartates, valerates, oleates, laurates, borates, benzoates, lactates, phosphates, toluenesulfonates (p-toluenesulfonates), citrates, malates, maleates, fumarates, succinates, tartrates, naphthalenesulfonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, benzenesulfonates, butyrates, camphorates, camphorsulfonates, cyclopentanepropionates, disglucurons, dodecyl sulfates, gluconate, glyceryl phosphates, heptarates, hexanoates, undecanoates, 2-hydroxyethanesulfonates, ethanesulfonates, etc. Representative inorganic salts can be formed from inorganic acids, such as sulfates, hydrogen sulfates, hemisulfates, hydrochlorides, chlorates, perchlorates, hydrobromates, and hydroiodates. Examples of basic salts include ammonium salts; alkali metal salts, such as sodium and potassium salts; alkaline earth metal salts, such as calcium and magnesium salts; organic basic salts, such as dicyclohexylamine, N-methyl-D-glucosamine, and phenylethylamine; and salts of amino acids such as arginine and lysine. These salts can be readily prepared using methods well known in the art.

[0132] In some embodiments, one or more of the compounds described herein are administered to a patient as prodrugs. A prodrug, as is well known in the art, refers to a compound administered in the form of a drug that is subsequently metabolized into a pharmacologically active form. Any compound that serves as a prodrug of a compound of the present invention is within the scope and spirit of the invention. Conventional methods for preparing suitable prodrugs of the present invention are described in textbooks, such as "Design of Prodrugs," edited by H. Bundgaard, Elsevier, 1985, the entire contents of which are incorporated herein by reference.

[0133] Compound Synthesis

[0134] Some of the compounds of this invention were synthesized according to the following reaction schemes, which are provided as examples only.

[0135] The aniline intermediate was synthesized as follows: starting with 4-nitrobenzene-1,2-diol (2-1), an alkylation reaction was carried out to protect the phenolic group, followed by reduction of the nitro group. The two fragments were then coupled using HATU, followed by debenzyl protection to obtain the target compound.

[0136] The biaryl compound was synthesized as follows: starting with pyridine-2-amine (1-5), an amide coupling reaction was carried out to generate a carboxylic acid intermediate (5-2). Further coupling with bromoaniline yielded intermediate (5-3), which was then coupled with a substituted phenylboronic acid via a Suzuki reaction to generate intermediate (5-4). Subsequently, demethylation yielded the target compound.

[0137] The compounds of this invention are referred to herein by the identifier code "HC" followed by 4 or 5 digits. The compound designated ZBBP-92 is a reference compound and has the following chemical structure:

[0138] .

[0139] synthesis N -(3,4-Dihydroxyphenyl)- N -(6-(methoxy- d 3 pyridin-2-yl)cyclopropane-1,1-dicarboxamide

[0140]

[0141] Synthesis of 4-hydroxy-2'-(3-((6-(methoxy- d 3) Pyridin-2-yl)amino)-3-oxopropionyl)-[1,1'-biphenyl]-3-carboxylic acid

[0142]

[0143] Discussion on the determination of thioflavone-T

[0144] The structures of HC8766 and HC8768 and their activities in the thioflavone-T assay (see [link to relevant documentation]). Figure 1-3 The comparison suggests that there are two (possibly three) differences between these two structures that could affect their ability to significantly enhance the inhibition of amylin fibrillation:

[0145] 1) The spacing (length) between two NH groups in the central chain should be as short as possible (but not too short).

[0146] 2) The orientation (cis or trans) of these two NH groups (and C=O group) is more conducive to inhibiting fiber formation.

[0147] 3) The difference in the position of the OH group (relative to the NH group) on the left benzene ring can affect the structure-activity relationship of the two compounds.

[0148] Results of HC8761 ( Figure 5 This highlights the importance of the spacing between NH groups. HC8761 exhibits similar fiber formation inhibition to ZBBP92 at a molar ratio of 1:1 for hA: compounds, but its inhibitory effect is not as great as that of HC8768 at the same molar ratio.

[0149] The orientation of the NH and C=O groups in HC8761 is the same as that in HC8768, but there are two additional methyl groups (or ethyl groups) in the backbone chain, which makes the spacing between the NH groups larger.

[0150] The structure of HC87610 is very similar to that of HC8761, but an oxygen moiety is added to the central methyl group of its backbone. While HC87610 does exhibit inhibition of fiber formation (see...), Figure 4 However, its inhibition level was lower than that of ZBBP92 and HC8761, which may indicate that the conformational flexibility around the central oxygen part has changed, which reduces its ability to bind and inhibit fiber formation.

[0151] HC8762, HC8765, and HC8769 each contain a carboxylic acid group at the meta position relative to the attached NH group on the left-side benzene ring. None of these compounds showed good inhibition of fiber formation in thioflavin T assays.

[0152] HC9532, HC9533, and HC9534 are isomers, with the biphenyl hydroxycarboxylic acid moiety of each compound attached to the NH group at different positions. This positional difference affects the inhibition of hA fiber formation, which may be due to steric hindrance.

[0153] The structure of HC87682 differs from that of HC8768 in that the hydroxyl group on the left-hand benzene ring is replaced by an ester group. This reduces solubility (under 70% aqueous conditions) and may affect the observed activity. Furthermore, the presence of two longer groups on the benzene chain may also affect its binding affinity to hA.

[0154] Cytotoxicity

[0155] Example 17 showed that compound HC87682 was cytotoxic when cultured alone with RINm5F and CM cells. Compounds HC956, HC9532, HC9533, and HC9534 were not cytotoxic when used alone, and anti-hA-mediated β-cell death was analyzed.

[0156] Cell death prevention assays showed that HC956 did not possess cell death prevention activity. However, HC9532, HC9533, and HC9534 exhibited significant preventive activity against hA-mediated β-cell death, with HC9534 showing the highest level of preventive activity among the compounds tested.

[0157] Human amylin and amylin amyloidosis

[0158] In some embodiments, the present invention relates to compounds capable of preventing or reversing human amylin (hA) misfolding, aggregation, and / or amylin plaque amyloid formation. In some embodiments, the present invention relates to compounds capable of preventing and / or reversing the conversion of soluble human amylin to insoluble human amylin and / or capable of preventing and inhibiting and / or reversing the formation of cytotoxic amylin fibrils.

[0159] Human amylin, also known as islet amylin amyloid polypeptide (IAPP), is a 37-residue peptide hormone produced and secreted by the β cells of Langerhans islets in the human pancreas. In vivo production of hA involves converting an 89-residue sequence into a 67-amino acid pro-amyloid sequence (proIAPP). This pro-amyloid sequence is then post-translational modified to generate hA. Human amylin is produced normally in the body and works with insulin to control blood glucose levels.

[0160] It will be clear to those skilled in the art that the type of protein is generally associated with the site of amyloidosis in the body. Therefore, there are multiple types of amyloid proteins, each classified according to the proteins it contains, and the specific type of amyloidosis is classified according to the site of amyloid protein accumulation.

[0161] The deposition of amyloid proteins in the body typically involves the accumulation of insoluble amyloid fibrils containing misfolded proteins.

[0162] Human amylin is a misfoldable protein that leads to the conversion of soluble amylin monomers and oligomers into insoluble amylin-amyloid fibrils. When these insoluble aggregates are present in the islets of Langerhans, they can cause β-cell death and other cytotoxic effects, the mechanisms of which are not fully understood. Not wanting to be bound by theory, the inventors believe that the aggregation of islet amylin-amyloid proteins into islet amylin-amyloid fibrils and / or the misfolding of human amylin (hA) contributes at least in part to the toxicity observed in amylin-amyloid-related diseases such as type 2 diabetes and is believed to play a role in the pathological processes of these diseases.

[0163] Destruction of amyloid-beta protein by pancreatic amyloid

[0164] The inventors have discovered that the compounds described herein can disrupt the formation of amyloid proteins, such as islet amyloid proteins.

[0165] Disruption of amylin amyloidosis can include both prior and subsequent disruption. Prior disruption refers to any part of the process by which the compounds described herein interfere with the conversion of soluble monomers and oligomers (e.g., amylin monomers and oligomers) into insoluble amylin amyloid protein. Subsequent disruption refers to the depolymerization of already formed insoluble amylin amyloid protein (e.g., islet amylin amyloid), which may include the redissolution of the constituent proteins.

[0166] Disruption of insoluble amylin-amyloid protein can occur directly or indirectly. Direct disruption involves the binding of one or more compounds described herein or their metabolites to a portion of a protein monomer or oligomer (e.g., human amylin monomer or oligomer) or a portion of a pre-formed aggregate, physically preventing further aggregation and / or reversing aggregation, for example by weakening the interactions between monomers and / or oligomers within the aggregate. In contrast, indirect disruption involves preventing the formation of filaments or aggregates through mechanisms that do not require direct contact between the compound and the protein, such as by altering local conditions to present the protein in a soluble rather than insoluble form, or by promoting a non-aggregating conformation of the monomer or oligomer.

[0167] In some embodiments of the invention, the direct disruption of aggregates (e.g., islet aggregates) by the compounds described herein involves binding via covalent interactions; in other embodiments, binding involves binding via non-covalent interactions. In various embodiments, the compounds described herein interact with protein monomers or oligomers constituting amyloid protein or with amyloid protein itself via one or more covalent interactions and / or a combination of one or more non-covalent interactions (e.g., van der Waals forces).

[0168] For example, not wanting to be bound by theory, the inventors believe that, in various embodiments, the compounds described herein covalently bind to the monomeric form of proteins, such as hA. Again, not wanting to be bound by theory, this is believed to stabilize the conformation of the monomeric form and / or the soluble oligomer (non-toxic form), directly inhibiting the formation of amyloid-beta protein.

[0169] It can also indirectly cause the destruction of amylin amyloid protein, for example, if the compound does not bind directly to the protein monomers or oligomers that make up amylin amyloid protein or to amylin amyloid protein itself, but instead affects other parts of the amylin amyloid denaturation process that prevent or inhibit the formation of further aggregates or cause the depolymerization of pre-formed aggregates.

[0170] Unwilling to be bound by theory, the inventors believe that one of the mechanisms of insoluble islet amylin-amyloid formation begins with the aggregation of proamyloid, which subsequently serves as the initiation site for the deposition of insoluble islet amylin-amyloid fibrils. In some embodiments, therefore, the destruction of islet amylin-amyloid occurs by directly or indirectly disrupting the aggregation of proamyloid.

[0171] Regarding islet amyloid-acid amyloid protein, experiments have shown that under normal conditions, hA may be in a suitably disordered state, primarily composed of randomly coiled structures. Under certain conditions typically associated with pathogenic processes, hA is expected to misfold, producing β-sheets that stack to form highly structured aggregates containing multiple β-sheets. In some embodiments, disruption of islet amyloid-acid amyloid protein involves preventing or reversing hA misfolding. Misfolding of the protein components of amyloid-acid amyloid protein is also believed to play a role in other diseases associated with amyloidosis.

[0172] In several embodiments, the compounds described herein disrupt amylin-amyloid aggregates through one or more mechanisms selected from: preventing, inhibiting, and / or reversing the formation of amylin-amyloid fibrils; preventing and / or reversing the conversion of soluble human amylin to insoluble human amylin; and preventing, inhibiting, and / or reversing the formation of cytotoxic amylin fibrils. In some embodiments, the compounds described herein disrupt the formation of amylin-amyloid fibrils by preventing, inhibiting, and / or reversing protein misfolding. For example, the compounds described herein disrupt the formation of islet amylin-amyloid fibrils by preventing, reversing, and / or reversing hA misfolding.

[0173] In several embodiments, the compounds described herein prevent, inhibit, and / or reverse protein misfolding and / or prevent, inhibit, and / or reverse protein aggregation. Not wishing to be bound by theory, the inventors believe that the aggregation potential of hA is facilitated by the misfolding of amylin into structured β-sheets.

[0174] Effects on disease state

[0175] Amylin amyloid is believed to play a role in the pathogenesis of a variety of diseases. Amylin amyloid-related diseases occur in a variety of animals, including mammals such as humans.

[0176] The inventors have discovered that the compounds described herein can prevent, inhibit, and / or reverse, for example, the formation of amylin-amyloid fibrils, Langerhans island β-cell death, the conversion of soluble human amylin to insoluble human amylin, and the formation of cytotoxic oligomers. Not wishing to be bound by theory, the inventors believe that these processes are associated with the occurrence and / or progression of one or more amylin-amyloid-related diseases such as type 2 diabetes.

[0177] The term "block" refers to the prevention of a process that has not yet begun, such as amylin-amyloid fibrillation, Langerhans island β-cell death, the conversion of soluble human amylin to insoluble human amylin, and the formation of cytotoxic oligomers. In some embodiments, this block lasts for a period of time—for example, as long as the concentration of one or more compounds described herein (e.g., the local concentration of hA) remains above a certain threshold. It is understood that in such embodiments, "block" does not imply permanent block. The term "inhibition" as used herein is similar to "block," but refers to the prevention of a process that has already begun, such as amylin-amyloid fibrillation, Langerhans island β-cell death, the conversion of soluble human amylin to insoluble human amylin, and the formation of cytotoxic oligomers.

[0178] Inhibiting, suppressing, or reversing processes associated with the development of diseases related to one or more amylin-amyloid proteins can take many forms. For example, in some embodiments, inhibiting, suppressing, or reversing processes such as amylin-amyloid fibrillation, Langerhans island β-cell death, conversion of soluble human amylin to insoluble human amylin, and formation of cytotoxic oligomers can delay disease progression and improve the quality of life of patients with said diseases.

[0179] In several implementations, preventing, inhibiting, or reversing processes associated with the development of one or more amylin-amyloid proteins manifests as improved survival. For example, in some implementations, preventing, inhibiting, or reversing processes associated with hA amylin-amyloid formation will result in improved pancreatic β-cell survival.

[0180] The term "treatment" refers to the suppression or prevention of the development of amylin-amyloid-related disease and / or the reduction, alleviation, or reversal of amylin-amyloid-related disease or one or more of its side effects. Methods for evaluating treatment, including methods for assessing the suppression, prevention, reduction, alleviation, and / or reversal of the disease state, are known and readily apparent to those skilled in the art. The term "reversal" refers to the return of amylin-amyloid-related disease to a previous or milder stage of development. The terms "relief" and "reversal" are interpreted in a similar manner.

[0181] Application and formulation

[0182] The term "administration" refers to providing a patient with a therapeutically effective amount of a compound using one or more methods of administration known in the art. These methods include administration via oral, sublingual, intravenous, subcutaneous, transdermal, intramuscular, intradermal, intrathecal, epidural, intraocular, intracranial, inhalation, rectal, vaginal, and other methods.

[0183] In an exemplary embodiment, one or more active ingredients may be administered orally.

[0184] Those skilled in the art will appreciate that the formulation of the compounds described herein will depend on the method of administration. For example, in some embodiments, the compounds described herein are formulated as creams, lotions, tablets, capsules, microspheres, dispersible powders, granules, suppositories, syrups, elixirs, lozenges, injectable solutions, sterile aqueous or non-aqueous solutions, suspensions or emulsions, patches, etc. For example, in many embodiments, the compounds described herein are formulated as tablets, capsules, microspheres, dispersible powders, granules, syrups, suspensions, or emulsions.

[0185] In exemplary embodiments, the compounds described herein are formulated into solid dosage forms, such as tablets, capsules, or microspheres.

[0186] Formulations suitable for a particular method of administration will be obvious to those skilled in the art.

[0187] Those skilled in the art will understand that the route of administration and the nature of the pharmaceutically acceptable carrier depend on the nature of the disease and the mammal being treated. It is believed that those skilled in the art can readily determine the specific carrier or delivery system and the choice of route of administration. In preparing any formulation containing an active compound, care should be taken to ensure that the activity of the compound is not destroyed during preparation and that the compound can reach its site of action without degradation. In some cases, it may be necessary to use methods known in the art, such as microencapsulation, to protect the compound. Similarly, the chosen routes of administration should be those that allow the compound to reach its site of action.

[0188] Those skilled in the art can readily determine suitable formulations of the compounds of the present invention using conventional methods. Determining the preferred pH range and suitable excipients (e.g., antioxidants) is standard practice in the art. Buffer systems are conventionally used to provide the desired pH range, including carboxylate buffers such as acetate, citrate, lactate, and succinate. A variety of antioxidants can be used in such formulations, including phenolic compounds such as BHT or vitamin E, reducing agents such as methionine or sulfites, and metal chelating agents such as EDTA.

[0189] The compounds of the present invention, or pharmaceutically acceptable salts thereof, can be prepared into parenteral dosage forms, including those suitable for intravenous, intrathecal, intracerebral, or epidural delivery. Injectable dosage forms include sterile injectable solutions or dispersions, as well as sterile powders for the provisional preparation of sterile injectable solutions. They should be stable under manufacturing and storage conditions, free from reduction or oxidation, and from contamination by microorganisms such as bacteria or fungi.

[0190] The solvent or dispersion medium of the injectable solution or dispersion may contain any conventional solvent or carrier system used for the active compound, and may contain, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils. Appropriate flowability can be maintained by, for example, coating with lecithin, by maintaining the desired particle size in the dispersion, and by using surfactants. Where necessary, the action of microorganisms can be prevented by adding various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many cases, it is preferable to add substances that adjust osmotic pressure, such as sugars or sodium chloride. In many embodiments, the injectable formulation is isotonic with blood. The absorption of the injectable composition can be prolonged by using components that delay absorption, such as aluminum monostearate and gelatin. The injectable form of the drug can be delivered via any suitable route, including intravenous, intramuscular, intracerebral, intrathecal, epidural injection, or infusion.

[0191] Sterile injectable solutions are prepared by incorporating the desired amount of the active compound, as appropriate, with various other components, such as those listed above, into a suitable solvent, followed by sterilization by filtration. Dispersions are typically prepared by adding various sterilized active ingredients to a sterile solvent containing a basic dispersion medium and other desired components listed above. For sterile powders used to prepare sterile injectable solutions, a preferred method is to vacuum-dry or freeze-dry a solution of the pre-sterile filtered active ingredient with any other desired components.

[0192] Other pharmaceutical dosage forms of the present invention include oral and enteral preparations, wherein the active compound may be formulated with an inert diluent or an absorbable food carrier, or may be encapsulated in hard or soft-shell gelatin capsules, or may be compressed into tablets, or may be directly incorporated into food. For oral therapeutic administration, the active compound may be mixed with excipients and used in the form of swallowable tablets, buccal or sublingual tablets, lozenges, capsules, elixirs, suspensions, syrups, waffle tablets, etc. The amount of the active compound in such therapeutic compositions is the amount to obtain an appropriate dosage.

[0193] Tablets, lozenges, pills, capsules, etc., may also contain the following ingredients: binders, such as gum arabic, corn starch, or gelatin; excipients, such as dicalcium phosphate; disintegrants, such as corn starch, potato starch, alginic acid, etc.; lubricants, such as magnesium stearate; and sweeteners, such as sucrose, lactose, or saccharin, may be added, or flavoring agents, such as peppermint, wintergreen oil, or cherry flavoring. When the dosage unit is in capsule form, it may also contain a liquid carrier in addition to the substances of the types mentioned above. Various other substances may be present as coatings or to otherwise alter the physical form of the dosage form. For example, tablets, pills, or capsules may be coated with shellac, sugar, or both. Syrups or elixirs may contain active compounds, sucrose (as a sweetener), methylparaben and propylparaben (as preservatives), colorings, and flavoring agents (such as cherry or orange flavoring). Of course, any raw materials used to prepare any dosage unit form should be pharmaceutically pure and substantially non-toxic at the amounts used. In addition, active compounds can be incorporated into sustained-release formulations and formulations, including those that specifically deliver the active compound to specific regions of the intestine.

[0194] Liquid formulations can also be administered enterically via a gastric tube or esophagus. Enteric formulations can be prepared as suppositories by mixing with a suitable matrix (e.g., an emulsified matrix or a water-soluble matrix). The compounds of the present invention can also (but are not required to) be administered topically, intranasally, intravaginally, intraocularly, or intraocularly.

[0195] The invention also includes any other forms suitable for application, such as topical applications like creams, lotions, and gels, or compositions suitable for inhalation or intranasal delivery, such as solutions, dry powders, suspensions, or emulsions.

[0196] The compounds of the present invention can be administered by inhalation or as an aerosol spray from a pressurized dispenser or container containing a propellant, such as carbon dioxide gas, dichlorodifluoromethane, nitrogen, propane, or other suitable gases or combinations thereof. The compounds of the present invention can also be administered using a nebulizer.

[0197] Pharmaceutically acceptable carriers and / or diluents include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonics, and absorption delay agents. The use of such media and reagents for the use of pharmaceutically active substances is well known in the art. Unless any conventional media or reagent is incompatible with the active ingredient, its use in therapeutic compositions should be considered. Additional active ingredients may also be incorporated into the composition.

[0198] Formulating the composition into unit dosage forms is particularly advantageous for ease of administration and uniformity of dosage. A unit dosage form refers to a physically independent unit of a single dose suitable for a mammalian subject to be treated; each unit contains a predetermined amount of the active substance, calculated to produce the desired therapeutic effect when used in combination with a desired pharmaceutically acceptable carrier. The specifications of the novel unit dosage forms of the present invention depend on and are directly dependent on: (a) the unique properties of the active substance and the specific therapeutic effect to be achieved, and (b) the inherent limitations of prior art formulations of active substances for treating diseases in living patients suffering from disease states in which physical health is impaired, as detailed herein.

[0199] For ease of administration and effectiveness, the main active ingredient may be formulated in a therapeutically effective amount together with a suitable pharmaceutically acceptable carrier in a unit dosage form. A unit dosage form may, for example, contain 0.25 μg to approximately 2000 mg of the main active compound. Proportionally, the active compound may be present in an amount of approximately 0.25 μg to approximately 2000 mg / mL of the carrier. For compositions containing supplemental active ingredients, the dosage is determined with reference to the commonly used dosage and method of administration of said ingredient.

[0200] In some embodiments of the invention, a therapeutically effective amount refers to a dose sufficient to produce one or more of the following compounds: prevention, inhibition, or reversal of amylin-amyloid fibrillation, Langerhans island β-cell death, conversion of soluble human amylin to insoluble human amylin, and formation of cytotoxic oligomers.

[0201] In some cases, the therapeutically effective dose of a compound will be influenced by a number of factors and can be adjusted accordingly. For example, the therapeutically effective dose can be affected by the patient's weight, metabolic capacity, and synergistic effects between the combinations of active ingredients administered. The dose that can be administered to a patient can also be affected by other factors, such as interactions with other medications the patient is receiving and the severity / tolerance to any side effects of the administered compound.

[0202] In some implementation schemes, the expected outcomes of an effective therapeutic dose include slowing disease progression, improving patients’ quality of life, and / or increasing survival rates.

[0203] In various implementation schemes, the intended outcome of the therapeutically effective dose is to improve one or more symptoms associated with amyloid-related diseases, such as reduced renal function loss, heart failure, obstructive sleep apnea, dysphagia, or synovitis.

[0204] In various implementation schemes, the expected outcome of the therapeutically effective dose is improvement of one or more symptoms associated with type 2 diabetes, such as weight loss, polyuria, polydipsia, polyphagia, blurred vision, headache, fatigue, or diabetic dermadromes, as well as signs such as decreased blood glucose levels and HbA1C.

[0205] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or diluent.

[0206] The compounds described herein may be administered alone in some embodiments, and in other embodiments in combination with other compounds described herein or with other therapeutic agents, or both. This combination allows the compounds to be administered separately, sequentially, or simultaneously with other active ingredients. The combination may be provided as a pharmaceutical composition.

[0207] For example, in some implementations, two or more compounds are administered to a patient together. When the compounds are administered to a patient in combination, the dose of each individual compound may be less than a therapeutically effective amount, such that the combined dose of the two or more compounds is equal to or greater than a therapeutically effective amount.

[0208] Furthermore, in some embodiments, one or more compounds are administered to a patient together with one or more additional substances. These additional substances may, for example, be drugs for treating, inhibiting, and / or reversing amylin-amyloid fibrillation, Langerhans island β-cell death, the conversion of soluble human amylin to insoluble human amylin, and the formation of cytotoxic oligomers. In some embodiments, the one or more additional substances may be compounds for the prevention or treatment of amylin-amyloid-related diseases such as type 2 diabetes.

[0209] For example, in some embodiments, the one or more additional substances are selected from blood glucose regulators, such as insulin, insulin analogs or derivatives, Symlin, GLP-agonists, metformin, sulfonylurea drugs, thiazolidinediones, SGLT2 inhibitors, and selective dipeptidyl peptidase (DPP-IV) inhibitors.

[0210] In various instances, the GLP-1 agonist is exenatide, liraglutide, lixisenatide, albiglutide, or dulaglutide, or any combination of two or more thereof.

[0211] In various instances, selective dipeptidyl peptidase (DPP-IV) inhibitors are selected from sitagliptin, vildagliptin, saxagliptin, linagliptin, anagliptin, tenegliptin, alogliptin, trelagliptin, gemigliptin, dutogliptin, and omaligliptin.

[0212] In one instance, the selective dipeptidyl peptidase (DPP-IV) inhibitor is selected from alogliptin, linagliptin, saxagliptin, sitagliptin, Nesina, Tradjenta, Onglyza, and Januvia.

[0213] In some implementations, the one or more additional substances may be, for example, drugs that reduce the side effects associated with the compounds described herein.

[0214] In several embodiments, one or more compounds may be administered to a patient together with one or more additional substances selected from quinacrine, tetracycline, and doxycycline.

[0215] In several embodiments for in vitro use, one or more compounds may be applied to or in contact with an in vitro sample together with one or more additional substances selected from anthracene, phenanthrene, quinacrine, neutral red, chlorpromazine, acridine, acridine orange, methylene blue, phenadiazine, phenothiazine, tetracycline, doxycycline, Congo red, pyrene, β, benzo[a]anthracene, benzo[m]anthracene, benzo[c]phenanthrene, and tetraphenylene.

[0216] When one or more compounds are administered in combination with one or more additional substances, the dosage of each compound may be less than the dosage of the compound administered alone.

[0217] In some implementations, when one or more compounds are administered in combination with one or more additional substances, the dosage of each compound may be greater than the dosage when the compound is administered alone. This may occur, for example, if one or more additional substances reduce side effects, thereby allowing for tolerance to higher doses.

[0218] Example

[0219] Example 1: Synthesis of N-(3,4-dihydroxyphenyl)-N-(6-(methoxy-d3)pyridin-2-yl)cyclopropane-1,1-dicarboxamide (HC863O)

[0220]

[0221] Step 1. Synthesis of (((4-nitro-1,2-phenylene)bis(oxy))bis(methylene))diphenyl(2-2)

[0222] Sodium hydroxide (3.87 g, 96.7 mmol) and benzyl bromide (16.54 g, 96.7 mmol) were added to a solution of 4-nitrobenzene-1,2-diol (5.0 g, 32.2 mmol) in DMF. The resulting solution was stirred at 80°C for 8 h. After stirring, the reaction mixture was cooled to room temperature. The mixture was then quenched by slowly adding 200 mL of water. The resulting solid was filtered, washed with water, and dried under reduced pressure to give the title compound as a white solid (7.6 g, 70%). 1 H NMR (500 MHz, DMSO) δ 7.88 (d, J = 9.4 Hz, 1H), 7.86 – 7.84 (m, 1H), 7.45 (d, J = 7.6 Hz, 4H), 7.37 (d, J = 8.2 Hz, 4H), 7.33 (t, J = 6.0 Hz, 2H), 7.27 (d, J = 9.0 Hz,1H), 5.29 (s, 2H), 5.25 (s, 2H).

[0223] Step 2. Synthesis of 3,4-bis(benzyloxy)aniline (2-3)

[0224] Iron powder (2.0 g, 36 mmol) and ammonium chloride (1.9 g, 36 mmol) were added to a solution of (((4-nitro-1,2-phenylene)bis(oxy))bis(methylene))diphenyl (3.0 g, 9 mmol) in 10 mL of ethanol and 10 mL of water. The resulting solution was heated to 90°C and stirred for 2 hours. After completion, the mixture was cooled to room temperature and filtered through a diatomaceous earth mat. The resulting green leaf was extracted with ethyl acetate. The organic layer was separated and dried over sodium sulfate. Volatile components were removed under reduced pressure, and the residue was purified on a silica gel column using petroleum ether / ethyl acetate = 3:1 as eluent. The pure fraction was evaporated to dryness to give the title compound as a yellow solid (2.30 g, 84%). 1H NMR (500 MHz, DMSO- d 6 )) δ 7.42 – 7.33 (m,10H), 6.71 (d, J = 8.5 Hz, 1H), 6.37 (s, 1H), 6.07 (d, J = 8.5 Hz, 1H), 5.10 (s, 2H), 5.01 (s, 2H), 4.90 (s, 2H).

[0225] Step 3. Synthesis of 1-((6-(methyl- d 3 2-pyridinyl carbamoyl cyclopropane-1-carboxylic acid methyl ester (2-4)

[0226] HOBT (0.91 g, 6.8 mmol) and EDCI (1.29 g, 6.8 mmol) were added to a solution of 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid (0.97 g, 6.8 mmol) in 5 mL of DCM at room temperature. The resulting solution was stirred for 10 minutes. Then, 6-(methoxy- d 3 Pyridine-2-amine (1-5, 0.5 g, 4.5 mmol). The reaction mixture was stirred for another 4 hours. After completion, the reactants were diluted with water and extracted with DCM. The organic layer was separated and dried over sodium sulfate. Volatile components were removed under reduced pressure, and the residue was purified on a silica gel column using petroleum ether / dichloromethane = 1:1 as eluent. The pure fraction was evaporated to dryness to give the title compound as a brown solid (0.77 g, 72%). 1 H NMR (400 MHz, DMSO- d 6 )) δ 10.88 (s, 1H), 7.69 (t, J = 7.8 Hz, 1H), 7.63 (d, J = 7.1 Hz, 1H), 6.54 (d, J = 7.8 Hz, 1H), 3.68 (s, 3H), 1.52 (s, 4H).

[0227] Step 4. Synthesis of 1-((6-(methyl- d 3 )pyridin-2-yl)carbamoyl)cyclopropane-1-carboxylic acid (2-5)

[0228] At room temperature to 1-((6-(methyl-d 3 2-(pyridin-2-yl)carbamoyl)cyclopropane-1-carboxylic acid methyl ester (2-4, 0.77 g, 6.8 mmol) was dissolved in 5 mL THF and 5 mL water, and sodium hydroxide (0.39 g, 9.7 mmol) was added. The resulting solution was stirred for 1 hour. After completion, the reactants were diluted with water, acidified with hydrochloric acid, and extracted with DCM. The organic layer was separated and dried over sodium sulfate. The volatile components were removed to give the title compound as a yellow solid (0.68 g, 94%). 1 H NMR (500 MHz, DMSO-) d 6 ) δ 11.20 (s, 1H), 7.66 (td, J = 7.9, 2.5 Hz, 1H), 7.61 (d, J = 8.1 Hz, 1H), 6.51 (dd, J = 7.9, 2.5 Hz, 1H), 1.52 (s, 2H), 1.51 (s, 2H).

[0229] Step 5. Synthesis N -(3,4-bis(benzyloxy)phenyl) -N -(6-(methoxy- d 3 )pyridin-2-yl)cyclopropane-1,1-dicarboxamide (2-6)

[0230] To 1-((6-(methyl- d 3 2-pyridin-2-yl)carbamoyl)cyclopropane-1-carboxylic acid (2-5, 0.37 g, 1.6 mmol) was added to a solution in 5 mL DMF, along with HATU (0.93 g, 2.5 mmol) and triethylamine (0.33 g, 3.3 mmol). The resulting solution was stirred for 10 minutes. Then, 3,4-bis(benzyloxy)aniline (2-3, 0.5 g, 1.6 mmol) was added in a single batch. The reaction mixture was stirred for another 4 hours. After completion, the reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was separated, washed with brine, and dried over sodium sulfate. The volatile components were removed, yielding a yellow solid, which was used for the next step without further purification.

[0231] Step 6. Synthesis N -(3,4-Dihydroxyphenyl)- N -(6-(methoxy- d 3)pyridin-2-yl)cyclopropane-1,1-dicarboxamide (HC863O)

[0232] Towards N -(3,4-bis(benzyloxy)phenyl) -N -(6-(methoxy- d 3 0.68 g (1 mmol) of pyridin-2-yl)cyclopropane-1,1-dicarboxamide was added to a solution of 10% palladium on carbon (wetted with about 55% water, 0.1 g) in 3 mL of dichloromethane and 3 mL of methanol. The resulting mixture was stirred at room temperature for 3 hours. After stirring, the mixture was filtered through a diatomaceous earth filter. The filtrate was concentrated under reduced pressure, and the residue was purified on a silica gel column using dichloromethane / methanol = 10:1 as the eluent. The purified fraction was evaporated to dryness to give the title compound as a white solid (0.35 g, 61% in both steps). 1 H NMR (400 MHz, DMSO) δ10.24 (s, 1H), 9.75 (s, 1H), 9.01 (d, J = 10.1 Hz, 1H), 8.66 (s, 1H), 7.67(dt, J = 15.2, 7.6 Hz, 2H), 7.10 (s, 1H), 6.78 (d, J = 8.7 Hz, 1H), 6.64 (d, J = 8.7 Hz, 1H), 6.53 (d, J = 7.5 Hz, 1H), 3.79 (t, J = 6.4 Hz, 1H), 3.64 (t, J = 6.5 Hz, 2H), 2.30 (q, J = 6.8 Hz, 2H). m / z (ES - [M - H] - = 345.2.

[0233] Example 2: Synthesis of 2-hydroxy-5-(1-((6-(methoxy-d3)pyridin-2-yl)carbamoyl)cyclopropane-1-carbamoylamino)benzoic acid (HC863C)

[0234]

[0235] By operating in a manner similar to that of Example 1, in step 1, 2-hydroxy-5-nitrobenzoic acid was used instead of 4-nitrobenzene-1,2-diol for synthesis. 1 H NMR (400 MHz, DMSO) δ 13.68 (s, 1H), 11.40 (s, 1H), 10.59 (s, 1H), 9.87 (s, 1H), 7.72 (d, J = 8.1 Hz, 1H), 7.68 (d, J = 7.9 Hz, 1H), 7.63 (d, J = 7.3 Hz, 1H), 7.36 (s, 1H), 7.16 (d, J = 8.7 Hz, 1H), 6.53 (d, J = 7.9 Hz, 1H), 1.58 (s, 2H), 1.52 (d, J = 7.3 Hz, 2H). m / z (ES - [M - H] - =373.2

[0236] Example 3: Synthesis of N 1 -(3,4-Dihydroxyphenyl)-N 5 -(6-(methoxy-d3)pyridin-2-yl)glutaramide (HC8761)

[0237]

[0238] By operating in a manner similar to that of Example 1, in step 3, 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid was synthesized instead of 5-methoxy-5-oxovalerate. 1 H NMR (400 MHz, DMSO) δ 10.22 (s, 1H), 9.52 (s,1H), 8.89 (s, 1H), 8.62 – 8.46 (m, 1H), 7.78 – 7.57 (m, 2H), 7.14 (s, 1H), 6.78 (d, J = 8.6 Hz, 1H), 6.60 (d, J = 7.7 Hz, 1H), 6.49 (d, J = 5.9 Hz, 1H),2.43 (s, 2H), 2.27 (t, J= 6.7 Hz, 2H), 1.90 – 1.81 (m, 2H). m / z (ES + [M +H] + = 348.8

[0239] Example 4: Synthesis of 2-hydroxy-5-(5-((6-(methoxy-d3)pyridin-2-yl)amino)-5-oxopentanoylamino)benzoic acid (HC8762)

[0240]

[0241] The synthesis was carried out by means of operation similar to that of Example 1, in step 3 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid was replaced by 5-methoxy-5-oxopentanoic acid and in step 1 4-nitrobenzene-1,2-diol was replaced by 2-hydroxy-5-nitrobenzoic acid. 1 H NMR (400 MHz, DMSO) δ 11.57 – 11.15 (m, 1H), 10.23 (s, 1H), 10.16 (s,1H), 7.78 – 7.57 (m, 3H), 7.37 (d, J = 1.8 Hz, 1H), 7.06 (dd, J = 8.7, 1.9Hz, 1H), 6.54 – 6.42 (m, 1H), 2.46 (t, J = 7.3 Hz, 2H), 2.40 (t, J = 7.4 Hz, 3H), 1.89 (p, J = 7.3 Hz, 2H). m / z (ES + [M + H] + = 376.8

[0242] Example 5: Synthesis of N 1 -(3,4-Dihydroxyphenyl)-N 6 -(6-(methoxy-d3)pyridin-2-yl)hexamethylenediamide (HC8764)

[0243]

[0244] By operating in a manner similar to that of Example 1, in step 3, 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid was synthesized instead of 6-methoxy-6-oxohexanoic acid. 1H NMR (400 MHz, DMSO) δ 10.19 (s, 1H), 9.50 (s,1H), 8.89 (s, 1H), 8.54 (s, 1H), 7.65 (d, J = 6.9 Hz, 2H), 7.13 (d, J = 2.4Hz, 1H), 6.76 (dd, J = 8.6, 2.4 Hz, 1H), 6.60 (d, J = 8.5 Hz, 1H), 6.53 –6.43 (m, 1H), 2.40 (d, J = 6.7 Hz, 2H), 2.25 (q, J = 6.7, 5.3 Hz, 2H), 1.59(p, J = 3.6 Hz, 4H). m / z (ES + [M + H] + = 362.8

[0245] Example 6: Synthesis of 2-hydroxy-5-(6-((6-(methoxy-d3)pyridin-2-yl)amino)-6-oxohexanoylamino)benzoic acid (HC8765)

[0246]

[0247] By operating in a manner similar to that of Example 1, the synthesis was carried out in step 3 by replacing 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid with 6-methoxy-6-oxohexanoic acid and in step 1 by replacing 4-nitrobenzene-1,2-diol with 2-hydroxy-5-nitrobenzoic acid. 1 H NMR (400 MHz, DMSO) δ 11.37 (s, 2H), 10.19 (s, 1H), 10.16 (s, 1H), 7.80– 7.52 (m, 3H), 7.36 (d, J = 2.0 Hz, 1H), 7.05 (dd, J = 8.7, 2.0 Hz, 1H), 6.54 – 6.38 (m, 1H), 2.43 (d, J = 6.2 Hz, 2H), 2.35 (d, J = 6.5 Hz, 2H), 1.61(h, J = 3.5 Hz, 4H).m / z (ES + [M + H] + = 412.8

[0248] Example 7: Synthesis of N 1 -(3,4-Dihydroxyphenyl)-N 4 -(6-(methoxy-d3)pyridin-2-yl)succinamide (HC8766)

[0249]

[0250] By operating in a manner similar to that of Example 1, in step 3, 4-methoxy-4-oxobutyric acid was used instead of 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid for synthesis. 1 H NMR (400 MHz, DMSO) δ 10.27 (s, 1H), 9.60 (s,1H), 8.88 (s, 1H), 8.53 (s, 1H), 7.64 (d, J = 5.4 Hz, 2H), 7.13 (d, J = 2.4Hz, 1H), 6.78 (dd, J = 8.5, 2.5 Hz, 1H), 6.61 (d, J = 8.5 Hz, 1H), 6.55 –6.40 (m, 1H), 2.70 (t, J = 7.0 Hz, 2H), 2.56 (t, J = 7.0 Hz, 2H). m / z (ES + [M + H] + = 334.8

[0251] Example 8: Synthesis of N 1 -(3,4-Dihydroxyphenyl)-N 3 6-(methoxy-d3)pyridin-2-yl)malonamide (HC8768)

[0252]

[0253] By operating in a manner similar to that of Example 1, in step 3, 3-methoxy-3-oxopropionic acid was used instead of 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid for synthesis. 1H NMR (400 MHz, DMSO) δ 10.40 (s, 1H), 9.80 (s,1H), 8.98 (s, 1H), 8.62 (s, 1H), 7.83 – 7.58 (m, 2H), 7.14 (d, J = 2.6 Hz, 1H), 6.79 (dd, J = 8.5, 2.5 Hz, 1H), 6.64 (d, J = 8.5 Hz, 1H), 6.57 – 6.49(m, 1H), 3.50 (d, J = 5.2 Hz, 2H). m / z (ES + [M + H] + = 320.8

[0254] Example 9: Synthesis of 2-hydroxy-5-(3-((6-(methoxy-d3)pyridin-2-yl)amino)-3-oxopropionylamino)benzoic acid (HC8769)

[0255]

[0256] By operating in a manner similar to that of Example 1, the synthesis was carried out in step 3 by replacing 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid with 3-methoxy-3-oxopropionic acid and in step 1 by replacing 4-nitrobenzene-1,2-diol with 2-hydroxy-5-nitrobenzoic acid. 1 H NMR (400 MHz, DMSO) δ 10.45 (s, 1H), 10.41 (s, 1H), 7.74 – 7.62 (m,3H), 7.32 (d, J = 2.1 Hz, 1H), 7.04 (dd, J = 8.7, 2.1 Hz, 1H), 6.59 – 6.50 (m, 1H), 3.61 (s, 2H). m / z (ES + [M + H] + = 348.9.

[0257] Example 10: Synthesis of N-(3,4-dihydroxyphenyl)-2-(2-((6-(methoxy-d3)pyridin-2-yl)amino)-2-oxoethoxy)acetamide (HC87610)

[0258]

[0259] By operating in a manner similar to that of Example 1, in step 3, 1-(methoxy-2-oxoethoxy)acetic acid was replaced with 2-(2-methoxycarbonyl)cyclopropane-1-carboxylic acid. 1 H NMR (400 MHz, DMSO) δ 10.17 (s, 1H), 9.74 (s, 1H), 8.98 (s, 1H), 8.65 (s, 1H), 7.71 (q, J = 11.3, 9.6 Hz, 2H), 7.19 (d, J = 2.4 Hz, 1H), 6.85 (dd, J = 8.5, 2.5 Hz, 1H), 6.65 (d, J = 8.5Hz, 1H), 6.56 (d, J = 8.0 Hz, 1H), 4.30 (s, 2H), 4.19 (s, 2H). m / z (ES + [M +H] + = 350.8.

[0260] Example 11: Synthesis of 4-(3-((6-(methoxy-d3)pyridin-2-yl)amino)-3-oxopropionylamino)-1,2-phenylene dipropionate (HC87682)

[0261]

[0262] At 0°C N 1 -(3,4-Dihydroxyphenyl)- N 3 -(6-(methoxy- d 3 0.1 g (0.3 mmol) of pyridin-2-yl)malonamide was added to a solution of triethylamine (0.09 g, 0.9 mmol) and propionyl chloride (0.09 g, 0.9 mmol) in 1 mL of THF. The resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was then concentrated under reduced pressure. The resulting residue was purified on a silica gel column using dichloromethane / methanol = 50:1 as the eluent. The purified fraction was evaporated to dryness to give the title compound as a white solid (0.082 g, 61%). 1H NMR (400 MHz, DMSO) δ 10.47 (s, 1H), 10.41 (s, 1H), 7.75 – 7.62 (m, 3H), 7.39 (dd, J = 8.8, 2.5 Hz, 1H), 7.21 (d, J =8.8 Hz, 1H), 6.58 – 6.49 (m, 1H), 3.58 (s, 2H), 2.58 (p, J = 7.5 Hz, 4H), 1.11 (t, J = 7.5 Hz, 6H). m / z (ES + [M + H] + = 433.1803.

[0263] Example 12: Synthesis of 4-hydroxy-2'-(3-((6-(methoxy-d3)pyridin-2-yl)amino)-3-oxopropionylamino)-[1,1'-biphenyl]-3-carboxylic acid (HC9532)

[0264]

[0265] Step 1. Synthesis of 3-((6-(methoxy- d 3) Pyridin-2-yl)amino)-3-oxopropionic acid (5-2)

[0266] At room temperature to 6-(methoxy- d 3) Triethylamine (0.80 g, 7.9 mmol) was added to a solution of pyridine-2-amine (0.50 g, 3.9 mmol) in 5 mL of DCM. The resulting solution was then cooled to 0°C, and methyl 3-chloro-3-oxopropionate (5-1 g, 0.81 g, 5.9 mmol) was added dropwise to the reaction mixture. The mixture was then warmed to room temperature and stirred for another hour. After completion, the mixture was quenched with water and extracted with dichloromethane. The organic phase was dried over sodium sulfate and concentrated under reduced pressure. The resulting residue was dissolved in 5 mL of tetrahydrofuran and 2 mL of water. Sodium hydroxide (0.47 g, 11.8 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for 2 hours. After completion, the mixture was quenched with 10 mL of 1N hydrochloric acid and extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to give the title compound as a pale yellow solid (0.79 g, 94%). 1H NMR (500 MHz, DMSO) δ 12.62 (s, 1H), 10.31 (s, 1H), 7.55 –7.52 (m, 1H), 7.51 – 7.39 (m, 2H), 3.34 (s, 2H). m / z (ES + [M + H] + = 214.1.

[0267] Step 2. Synthesis N 1 -(2-Bromophenyl)- N 3 -(6-(methoxy- d 3) Pyridin-2-yl)malonamide (5-3)

[0268] The synthesis was carried out in step 5 by replacing 2-5 with 5-2 and 3,4-bis(benzyloxy)aniline with 2-bromoaniline, using a similar procedure to that in Example 7. 1 H NMR (500 MHz, dmso) δ 10.48 (s, 1H), 9.85 (s, 1H), 7.78 (dd, J = 8.2, 1.6 Hz, 1H), 7.72 – 7.58 (m, 3H), 7.37 (td, J = 7.7, 1.5Hz, 1H), 7.11 (td, J = 7.7, 1.6 Hz, 1H), 6.56 – 6.47 (m, 1H), 3.67 (s, 2H). m / z (ES + [M + H] + = 367.1.

[0269] Step 3. Synthesis of 4-methoxy-2'-(3-((6-(methoxy-d3)pyridin-2-yl)amino)-3-oxopropionylamino)-[1,1'-biphenyl]-3-carboxylic acid (5-4)

[0270] In a nitrogen atmosphere, towards N 1 -(2-Bromophenyl)- N 3 -(6-(methoxy- d3) Pyridin-2-yl)malondiamide (5-3, 0.3 g, 0.8 mmol), 5-bromo-2-methoxybenzoic acid (0.24 g, 1.2 mmol), and sodium carbonate (0.26 g, 0.25 mmol) were added to a solution of 5 mL dioxane and 3 mL water, along with tetrakis(triphenylphosphine)palladium (0.09 g, 0.08 mmol). The reaction mixture was heated to 100°C and stirred under a nitrogen atmosphere for 3 hours. After completion, the reaction mixture was cooled to room temperature and filtered through a diatomaceous earth mat. The filtrate was then partitioned between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified on a silica gel column using dichloromethane / methanol = 15:1 as the eluent. The purified fraction was evaporated to dryness to give the title compound as a white solid (0.27 g, 75%). 1 H NMR (500 MHz, DMSO) δ12.57 (s, 1H), 10.34 (s, 1H), 9.56 (s, 1H), 7.70 – 7.58 (m, 4H), 7.53 (dd, J = 8.6, 2.4 Hz, 1H), 7.36 – 7.28 (m, 2H), 7.28 – 7.23 (m, 1H), 7.10 (d, J =8.6 Hz, 1H), 6.51 (d, J = 7.9 Hz, 1H), 3.79 (s, 3H), 3.44 (s, 2H). m / z (ES + [M + H] + = 439.3.

[0271] Step 4. Synthesis of 4-hydroxy-2'-(3-((6-(methoxy- d 3) Pyridin-2-yl)amino)-3-oxopropionyl)-[1,1'-biphenyl]-3-carboxylic acid (HC9532)

[0272] Tribromoborane (1 mol / L solution in DCM, 1.85 mL, 1.85 mmol) was added to a solution of 4-methoxy-2'-(3-((6-(methoxy-d3)pyridin-2-yl)amino)-3-oxopropionylamino)-[1,1'-biphenyl]-3-carboxylic acid (5-4, 0.27 g, 0.6 mmol) in 2 mL of DCM at -78°C. The reaction mixture was then allowed to cool to room temperature and stirred for another 2 hours. After completion, the reaction mixture was quenched with water. The resulting precipitate was then separated, washed with methanol, and dried under reduced pressure to give the title compound as a white solid (0.15 g, 57%). 1 H NMR (400 MHz, DMSO) δ 14.02 (s, 1H), 11.30 (s, 1H), 10.34 (s, 1H), 9.59 (s, 1H), 7.78 (d, J = 2.4 Hz, 1H), 7.69 (d, J = 7.9 Hz, 1H), 7.62 (d, J = 7.3 Hz, 2H), 7.55 (dd, J = 8.6, 2.4 Hz, 1H), 7.40 – 7.34 (m, 1H), 7.34 – 7.25 (m, 2H), 6.97 (d, J =8.5 Hz, 1H), 6.53 (d, J = 8.0 Hz, 1H), 3.46 (s, 2H). m / z (ES + [M + H] + =425.1536.

[0273] Example 13: Synthesis of 4-hydroxy-3'-(3-((6-(methoxy-d3)pyridin-2-yl)amino)-3-oxopropionylamino)-[1,1'-biphenyl]-3-carboxylic acid (HC9533)

[0274]

[0275] Similar to the procedure in Example 12, in step 2, 3-bromoaniline was used instead of 2-bromoaniline for synthesis. 1 HNMR (400 MHz, DMSO) δ 11.26 (s, 1H), 10.47 (s, 1H), 10.29 (s, 1H), 8.02 (d, J= 2.5 Hz, 1H), 7.91 – 7.85 (m, 1H), 7.81 (dd, J = 8.6, 2.5 Hz, 1H), 7.73 –7.63 (m, 2H), 7.64 – 7.46 (m, 2H), 7.40 (t, J = 7.8 Hz, 1H), 7.37 – 7.30 (m,1H), 7.08 (d, J = 8.6 Hz, 1H), 6.65 – 6.51 (m, 1H), 3.60 (s, 2H). m / z (ES + [M + H] + = 425.1537.

[0276] Example 14: Synthesis of 4-hydroxy-4'-(3-((6-(methoxy-d3)pyridin-2-yl)amino)-3-oxopropionylamino)-[1,1'-biphenyl]-3-carboxylic acid (HC9534)

[0277]

[0278] Similar to the procedure in Example 12, in step 2, 4-bromoaniline was used instead of 2-bromoaniline for synthesis. 1 HNMR (400 MHz, DMSO) δ 11.23 (s, 1H), 10.48 (s, 1H), 10.27 (s, 1H), 8.02 (d, J = 2.5 Hz, 1H), 7.83 (dd, J = 8.6, 2.4 Hz, 1H), 7.72 – 7.65 (m, 4H), 7.64 –7.57 (m, 2H), 7.05 (d, J = 8.6 Hz, 1H), 6.58 – 6.50 (m, 1H), 3.60 (s, 2H). m / z (ES + [M + H] + = 425.1539.

[0279] Example 15: Synthesis of N 1 -(3,4-bis(difluoromethoxy)phenyl)-N 3 6-(methoxy-d3)pyridin-2-yl)malonamide (HC956)

[0280]

[0281] Under vigorous stirring, towards N 1 -(3,4-Dihydroxyphenyl)- N 3 -(6-(methoxy- d 3 Pyridin-2-yl)malonamide (HC8768, Example 8, 0.05 g, 0.15 mmol) was added to a solution in 1 mL of DCM with an aqueous solution of KOH (20 wt%, 0.26 mL, 0.9 mmol). Then, a solution of TMSCF2Br (0.16 g, 0.8 mmol) in DCM (0.5 mL) was added at 0 °C, and the reaction mixture was allowed to warm to room temperature and stirred for another 30 minutes. After completion, the reaction mixture was partitioned between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified on a silica gel column using dichloromethane / methanol = 15:1 as the eluent. The purified fraction was evaporated to dryness to give the title compound as a white solid (0.053 g, 81%). 1 H NMR (400 MHz, DMSO) δ 10.47 (s, 1H), 10.46 (s, 1H), 7.74 (d, J = 2.4 Hz, 1H), 7.72 – 7.67 (m, 1H), 7.46 – 7.40 (m, 1H), 7.33 –7.30 (m, 1H), 7.20 – 7.11 (m, 1H), 7.03 – 6.91 (m, 1H), 6.54 (d, J = 7.6 Hz,1H), 6.01 – 5.82 (m, 1H), 3.58 (s, 2H). m / z (ES + [M + H] + = 421.1216.

[0282] Example 16: Inhibition of amylin fibrillation

[0283] This embodiment describes the use of the thioflavin-T assay to analyze representative compounds of formula (I) to assess the inhibition of amylin fibrillation mediated by the compounds.

[0284] All compounds were dissolved in an aqueous NaOH solution with pH > 10, and then the pH was adjusted to 7-8 with HCl. All compounds were 100% soluble and stable in solution for at least 24 hours under these conditions, except for HC8768, which began to precipitate after 30 minutes under pH > 10 conditions.

[0285] The ability of compounds to inhibit human amylin (hA) fibrillation was evaluated using the thioflavin-T assay, as demonstrated by J. Faitken et al. Suppression by polycyclic compounds of the conversion of human pancreatin into insoluble amyloid As described in Biochem J., 374 (2003), 779-784.

[0286] The interaction was determined using the thioflavone-T fluorescence method. When bound to hA fibers, thioflavone-T showed a significant increase in fluorescence, which could be quantified. If the compound was added to a solution of human amylin and interacted with amylin to inhibit or prevent amylin fiber formation, a quantifiable decrease in thioflavone-T fluorescence was observed. Therefore, the ability of the compound to inhibit human amylin fiber formation was tested using the thioflavone-T assay.

[0287] The effects of the compounds on human amylin fibrillation were determined by fluorescence spectroscopy using a Spectra-MAX Gemini XS fluorescence spectrophotometer (Molecular Devices Corporation, Sunnyvale, CA, USA). Maximum excitation and emission were set at 450 nm and 510 nm, respectively, with a 495 nm cutoff filter. The rate of human amylin fibrillation was determined in 10 mM Tris (pH 7.4) by monitoring thiosulfate-T fluorescence in the presence or absence of these compounds. Each experiment was performed in triplicate and independently at least twice. For each compound, inhibition was evaluated at two hA:compound ratios (1:1 and 1:0.1). Figure 1-9 The results show the hA:compound ratio of 1:1.

[0288] Figure 1 The display shows that HC8768 ( The inhibitory effect on fiber formation was significantly higher than that of ZBBP92. Both compounds showed inhibition of fibrosis compared to human amylin alone.

[0289] Figure 2 The display shows that HC87610 ( It inhibits fiber formation, but to a lesser extent than ZBBP92. HC8762 ( No inhibition of fiber formation was observed in HC8768. ) showed significantly higher than ZBBP92 ( ) inhibits fiber formation.

[0290] Figure 3 The display shows that HC8765 ( HC8764 showed very little inhibition of fiber formation. ) shows the same as ZBBP92 ( Similar fiber inhibition. HC8766 ( Its effect is weaker than that of ZBBP92 or HC8764.

[0291] Figure 4 The display shows that HC8769 ( It does not inhibit fiber formation. HC87610 ( It showed inhibition of fiber formation, but to a lesser extent than ZBBP92. ).

[0292] Figure 5 The display shows that HC8761 ( ) inhibits the formation of amylin fibrils to a degree similar to that of ZBBP92 ( )resemblance.

[0293] Figure 6 The display shows that HC9534 ( ) has the same characteristics as ZBBP92 ( It exhibits considerable fiber formation inhibition activity.

[0294] Figure 7 This shows that, compared to the activity level at 25 µM (see...), Figure 6 Compared to ), at 2.5 µM, HC9534 ( The inhibitory effect on fiber formation activity is stronger than that of ZBBP92. ).

[0295] Figure 8 The display shows that HC9532 ( HC9533 showed some, but weak, inhibition. It has a strong inhibitory effect on hA fiber formation, although it is not as strong as HC9534 ( ) or ZBBP92 ( That strong.

[0296] Figure 9 The display shows that HC87682 ( It has a strong inhibitory effect on hA fiber formation, although it is not as strong as HC9534 ( ) or ZBBP92 ( That strong.

[0297] Example 17: Effect on inhibiting human amylin-induced β-cell death

[0298] CM and RINm5F cells were seeded in 48-well plates and cultured overnight in RPMI medium (containing 5% and 10% FBS, respectively). Human amylin (hA) stock solution (500 μM) was freshly prepared with water and diluted with medium to a final concentration of 10 μM. Stock solutions (40 μM) for each compound were prepared with DMSO. Aliquots of hA medium (200 μL each) were premixed with each compound at a 1:10 molar ratio and incubated at room temperature for 2 hours. The mixture was then added to the cells and incubated overnight for 16–18 hours. Untreated control cells were treated similarly with water. The Cell Death Detection ELISA kit (Roche) was used, as described in (Zhang et al., Diabetes 57, 348-356, 2008; Zhang et al., J. Biol. Chem. (278, 52810-52819, 2003) to measure apoptotic cell death.

[0299] Figure 10 and Figure 11 The results shown represent the enrichment level of nucleosomes in the samples (cell lysates), expressed relative to the control (Co), with the control group set at 1. Values ​​are the mean ± SE of four independent experiments, with each experiment performed in duplicate. P <0.001 compared to the control; # P <0.05; ## P <0.01; ### P <0.001 compared to hA-treated cells.

Claims

1. Compound of formula (I): in: X is a C1-C5 alkyl, cyclopropyl, or C2-C4 dialkyl ether; A is a C1-C6 alkyl group that is absent or optionally substituted with a halogen, a C1-C3 alkoxy or a C1-C3 haloalkoxy, or A is a phenyl group; B is either absent or a C1-C6 alkyl group; R1 is independently selected from OH, halogen, C1-C3 alkoxy, C1-C3 deuterated alkoxy, C1-C3 haloalkoxy, C1-C3 deuterated haloalkoxy, CO2R3, OCO2R4 and O-glycosyl; R3 is H or a C1-C3 alkyl group; R4 is a C1-C3 alkyl group; and u is 1, 2, or 3; R2 is independently selected from OH, halogen, C1-C3 alkoxy, C1-C3 deuterated alkoxy, C1-C3 haloalkoxy, and C1-C3 deuterated alkoxy; and v is 1, 2, or 3; Or its pharmaceutically acceptable salt.

2. The compound as claimed in claim 1, wherein X is CH2.

3. The compound as claimed in claim 1, wherein X is (CH2)2, (CH2)3 or (CH2)4.

4. The compound as claimed in claim 1, wherein X is CH2-O-CH2.

5. The compound as claimed in claim 1, wherein X is cyclopropyl.

6. The compound as claimed in any one of claims 1-5, wherein A is a C1-C5 alkyl group substituted with OCH3, OCD3, OCH2F or F.

7. The compound as claimed in any one of claims 1-6, wherein R1 is OH, F, CO2H, OCH3, OCD3, OCF2H, OCOEt, OCO2Pr or O-glycosyl.

8. The compound as claimed in any one of claims 1-7, wherein R2 is OH, F, OCH3 or OCD3.

9. The compound as claimed in any one of claims 1-8, wherein u is 2 and R1 is OH.

10. The compound as claimed in any one of claims 1-8, wherein u is 2 and R1 are each O-glycosyl.

11. The compound claimed in any one of claims 1-8, wherein u is 2 and R1 is OCO2Pr.

12. The compound as claimed in any one of claims 1-8, wherein u is 2 and one R1 is OH and the other R1 is CO2H.

13. The compound as claimed in any one of claims 1-8, wherein u is 2 and one R1 is OH and the other R1 is OCF2H.

14. The compound as claimed in any one of claims 1-8, wherein u is 2 and one R1 is OH and the other R1 is F.

15. The compound claimed in any one of claims 1-14, wherein v is 1 and R2 is OCH3 or OCD3.

16. The compound claimed in any one of claims 1-14, wherein v is 2 and one R2 is OCH3 or OCD3 and the other R2 is F.

17. The compound as claimed in claim 1, selected from: 。 18. A pharmaceutical composition comprising a compound of any one of claims 1-17 and a pharmaceutically acceptable diluent, carrier, or excipient.

19. A method for treating or preventing diabetes, amyloid-associated disease, or Langerhans island β-cell death, the method comprising administering to a patient in need an effective amount of the compound of any one of claims 1-17.

20. A method for inhibiting, preventing, or reversing the formation of amylin amyloidosis or one or more amylin-amyloid fibrils or amylin-amyloid plaques, the method comprising contacting the amylin amyloidosis or one or more amylin-amyloid fibrils or amylin-amyloid plaques with an effective amount of the compound of any one of claims 1-17.

21. Use of the compound of any one of claims 1-17 in the preparation of a medicament, wherein the medicament is used to treat or prevent diabetes, amyloidosis-related diseases, or Langerhans island β-cell death.

22. A composition for treating or preventing diabetes, amyloid-associated diseases or Langerhans island β-cell death, comprising a compound of any one of claims 1-17.

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