C-glycosides as anti-inflammatory agents

Abstract

Provided herein are novel compounds and methods of using the compounds. The compounds include novel carbon analogs of pyranose derivatives, which have been found to have Toll-like receptor 4 (Tlr4) inhibitory activity. The methods are capable of treating infectious, inflammatory, and post-traumatic disorders.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the priority benefit of: U.S. Provisional Patent Application Serial Number 63 / 359,049 filed on July 7, 2022, U.S. Provisional Patent Application Serial Number 63 / 426,704 filed on November 18, 2022, and U.S. Provisional Patent Application Serial Number 63 / 443,835 filed on February 7, 2023), the contents of which are incorporated herein by reference in their entirety.

[0003] Fund Information

[0004] This disclosure was made with government support under Grant Nos. R01DK117186, R01DK083752, and R01GM078238 awarded by the National Institutes of Health. The government has certain rights in this invention. 1. Technical Field

[0005] The disclosed subject matter relates to novel carbon analogs of pyranose derivatives found to have Tlr4 inhibitory activity. The disclosed subject matter further provides methods of using the pyranose derivatives for treating infectious, inflammatory, and post-traumatic disorders. 2. Background technology

[0006] The innate immune receptor Toll-like receptor 4 ("Tlr4") has been implicated as a receptor on both hematopoietic and non-hematopoietic cells for endotoxin (lipopolysaccharide, "LPS"), as well as various endogenous molecules released in vivo during inflammatory or infectious disorders. Since Tlr4 is the most upstream receptor in the pro-inflammatory LPS signaling cascade, the use of inhibitors that antagonize Tl4 signaling can avoid the pitfalls associated with other cytokine inhibitors that act further downstream in the pathway and therefore have a lesser effect. 3. Summary of the invention

[0007] The purposes and advantages of the disclosed subject matter will be set forth in and apparent from the following description, and will be learned by practicing the disclosed subject matter. Other advantages of the disclosed subject matter will be realized and obtained by the devices particularly pointed out in the written description and claims hereof as well as in the accompanying drawings.

[0008] To achieve these and other advantages, and in accordance with the purposes of the disclosed subject matter, as specifically and broadly described, the disclosed subject matter includes carbon analogs of pyranose derivatives and methods of using the derivatives for treating infectious, inflammatory and post-traumatic diseases / disorders, and having Tlr4 signaling inhibitory activity. Certain compounds that can be used according to the present disclosure are listed in Table 1 below. In addition to the treatment methods, the disclosed subject matter further provides pharmaceutical compositions comprising the compounds and a suitable pharmaceutical carrier. Since Tlr4 is the most upstream receptor in the pro-inflammatory LPS signaling cascade, the treatments in the disclosed subject matter that inhibit or antagonize the action of Tlr4 can avoid the pitfalls associated with other cytokine inhibitors that act further downstream in the pathway and therefore have less specific effects (potentially non-critical).

[0009] In certain embodiments, the disclosed subject matter provides a compound of Formula I or a stereoisomer thereof:

[0010]

[0011] wherein X is O, Z is O or NH, Y is CH2 or CH(R1), R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2, or

[0012] wherein X is CH2, Z is O or NH, Y is O, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

[0013] The disclosed subject matter also provides a pharmaceutical composition comprising a compound of Formula I. In certain embodiments, the pharmaceutical composition is selected from the group consisting of coated particles, micelles, liposomes, tablets, capsules, sachets, suppositories, liquid pharmaceutical compositions, and combinations thereof. In certain embodiments, the pharmaceutical composition further comprises an antibiotic agent, a steroidal compound, or a nonsteroidal anti-inflammatory agent, or a combination thereof.

[0014] In certain embodiments, the compound of formula I is selected from the following structures:

[0015] 1-1

[0016] 1-2

[0017] 1-3

[0018] 1-4

[0019] 1-5

[0020] 1-6 (equivalent to 1-1),

[0021] 1-7

[0022] 1-8 (same as 1-2),

[0023] 1-9

[0024] 1-10 (equivalent to 1-17),

[0025] 1-11

[0026] 1-12

[0027] 1-13

[0028] 1-14

[0029] 1-15

[0030] 1-16

[0031] 1-17

[0032] 1-18

[0033] 1-19

[0034] 1-20 and combinations thereof.

[0035] The disclosed subject matter also provides compounds of Formula II or stereoisomers thereof:

[0036]

[0037] wherein Z is O or NH, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

[0038] The disclosed subject matter further provides a pharmaceutical composition comprising a compound of Formula II. In some of these embodiments, the pharmaceutical composition is selected from: coated particles, micelles, liposomes, tablets, capsules, sachets, suppositories, liquid pharmaceutical compositions, and combinations thereof. In some of these embodiments, the pharmaceutical composition further comprises: an antibiotic agent, a steroidal compound, or a non-steroidal anti-inflammatory agent, or a combination thereof.

[0039] According to some embodiments of the disclosed subject matter, the compound of formula II is selected from the following structures:

[0040] 2-1

[0041] 2-2

[0042] 2-3

[0043] 2-4

[0044] 2-5

[0045] 2-6 and combinations thereof.

[0046] In other embodiments, the disclosed subject matter provides methods of treating an infectious or inflammatory disorder comprising administering to a subject in need of such treatment an effective amount of a Toll-like receptor 4 inhibitor compound of Formula I or a stereoisomer thereof:

[0047]

[0048] wherein X is O, Z is O or NH, Y is CH2 or CH(R1), R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1 or NHR1, N(R1)2, or

[0049] wherein X is CH2, Z is O or NH, and Y is O, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

[0050] In another embodiment, the disclosed subject matter provides a method of treating an infectious or inflammatory disorder, the method comprising administering to a subject in need of such treatment an effective amount of a Toll-like receptor 4 inhibitor compound of Formula II, or a stereoisomer thereof:

[0051]

[0052] wherein Z is O or NH, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

[0053] In certain embodiments of the method, the subject suffers from one or more of the following: osteoarthritis, pancreatitis, metabolic syndrome, trauma-induced systemic inflammation, acute respiratory distress syndrome, and COVID-19-induced systemic inflammation. In a specific embodiment, the subject suffers from necrotizing enterocolitis.

[0054] According to the disclosed method, the Toll-like receptor 4 inhibitor is selected from:

[0055] 1-1

[0056] 1-2

[0057] 1-3

[0058] 1-4

[0059] 1-5

[0060] 1-6 (equivalent to 1-1),

[0061] 1-7

[0062] 1-8 (same as 1-2),

[0063] 1-9

[0064] 1-10 (equivalent to 1-17),

[0065] 1-11

[0066] 1-12

[0067] 1-13

[0068] 1-14

[0069] 1-15

[0070] 1-16

[0071] 1-17

[0072] 1-18

[0073] 1-19

[0074] 1-10 and combinations thereof.

[0075] In alternative embodiments, the Toll-like receptor 4 inhibitor is selected from:

[0076] 2-1

[0077] 2-2

[0078] 2-3

[0079] 2-4

[0080] 2-5

[0081] 2-6 and combinations thereof.

[0082] The disclosed subject matter also provides a method of treating a traumatic injury in a subject, the method comprising administering to a subject in need of such treatment an effective amount of a Toll-like receptor 4 inhibitor compound that reduces post-traumatic injury induced by Toll-like receptor 4. In certain embodiments, the traumatic injury is to an organ selected from the group consisting of: heart, liver, lung, kidney, intestine, brain, eye, and pancreas.

[0083] In certain embodiments, the compound is administered to treat organ rejection following transplantation.

[0084] In certain embodiments, the methods comprise administering a compound of Formula I or Formula II to prevent necrotizing enterocolitis in premature infants. 4. Description of the drawings

[0085] The following drawings are included to illustrate certain aspects of the present disclosure and should not be considered to be exclusive embodiments. The disclosed subject matter is capable of considerable modification, alteration, combination, and equivalents in form and function without departing from the scope of the present disclosure.

[0086] Figure 1Provided are the relative mRNA expression levels of Toll-like receptor 4 (Tlr4) in mouse enteroids cultures treated with: saline (Ctrl), lipopolysaccharide (LPS), or LPS with test compounds (C277, C278, C279, C280, C281, C282, C283, C284).

[0087] Figure 2 Presented are the relative mRNA expression levels of tumor necrosis factor (Tnf) in mouse enteroid cultures treated with: saline (Ctrl), lipopolysaccharide (LPS), or LPS with test compounds (C277, C278, C279, C280, C281, C282, C283, C284).

[0088] Figure 3 Relative mRNA expression levels of Toll-like receptor 4 (Tlr4) in the ileum of healthy control neonatal mice (Ctrl), mice with NEC, or mice with NEC treated with test compounds (C277, C278, C280, C281, C282, C283, C284) are provided. Statistical significance is indicated by p-values ​​of *p<0.05.

[0089] Figure 4 Relative mRNA expression levels of interleukin 6 (Il6) in the ileum of healthy control neonatal mice (Ctrl), mice with NEC, or mice with NEC treated with test compounds (C277, C278, C280, C281, C282, C283, C284) are provided. Statistical significance is indicated by p-values ​​of *p<0.05.

[0090] Figure 5 Relative mRNA expression levels of interleukin 1 beta (Il1b) in the ileum of healthy control neonatal mice (Ctrl), mice with NEC, or mice with NEC treated with test compounds (C277, C278, C280, C281, C282, C283, C284) are provided. Statistical significance is indicated by p-values ​​of *p<0.05.

[0091] Figure 6 Relative mRNA expression levels of tumor necrosis factor (Tnf) in the ileum of healthy control neonatal mice (Ctrl), mice with NEC, or mice with NEC treated with test compounds (C277, C278, C280, 5C281, C282, C283, C284) are provided. Statistical significance is indicated by p values ​​of *p<0.05, **p<0.01, ****p<0.0001.

[0092] Figure 7 Relative mRNA expression levels of lipocalin 2 (Lcn2) in the ileum of healthy control neonatal mice (Ctrl), mice with NEC, or mice with NEC treated with test compounds (C277, C278, C280, C281, C282, C283, C284) are provided. Statistical significance is indicated by p values ​​of *p<0.05, ***p<0.001.

[0093] Figure 8 Provided are the relative mRNA expression levels of Toll-like receptor 4 (Tlr4) in the ileum of neonatal mice treated with saline (Sal), or lipopolysaccharide (LPS), or LPS with test compounds (C277, C278, C279, C280, C281, C282, C283, C284). Statistical significance is indicated by p values ​​of *p<0.05, **p<0.01.

[0094] Fig. 9 Provided are the relative mRNA expression levels of interleukin 6 (Il6) in the ileum of newborn mice treated with saline (Sal), or lipopolysaccharide (LPS), or LPS with test compounds (C277, C278, C279, C280, C281, C282, C283, C284). Statistical significance is indicated by p values ​​of *p<0.05, **p<0.01, ****p<0.0001.

[0095] Fig.10 Provided are the relative mRNA expression levels of interleukin 1 beta (Il1b) in the ileum of neonatal mice treated with saline (Sal), or lipopolysaccharide (LPS), or LPS with test compounds (C277, C278, C279, C280, C281, C282, C283, C284). Statistical significance is indicated by p values ​​of **p<0.01, ***p<0.001, ****p<0.0001.

[0096] Fig.11 Relative mRNA expression levels of tumor necrosis factor (Tnf) in the ileum of neonatal mice treated with saline (Sal), or lipopolysaccharide (LPS), or LPS with test compounds (C277, C278, C279, C280, C281, C282, C283, C284) are provided. Statistical significance is indicated by p values ​​of ***p<0.001, ****p<0.0001.

[0097] Fig.12Provided are the relative mRNA expression levels of lipocalin 2 (Lcn2) in the ileum of neonatal mice treated with saline (Sal), or lipopolysaccharide (LPS), or LPS with test compounds (C277, C278, C279, C280, C281, C282, C283, C284). Statistical significance is indicated by p values ​​of *p<0.05, ***p<0.001, ****p<0.0001.

[0098] Fig.13 Representative images of hematoxylin and eosin (H&E) of non-NEC control, NEC, and NEC+C281 ileum tissue sections are provided. 5. Specific implementation methods

[0099] The terms used in this specification generally have their ordinary meaning in the art in the context of the present invention and in the specific context in which each term is used. Certain terms are discussed below or elsewhere in this specification to provide additional guidance to practitioners in describing the methods and compositions of the present invention and how to make and use them.

[0100] As used herein, when used with the term "comprising" in the claims and / or the specification, the expression "a, an" may refer to "one", but is also consistent with the meaning of "one or more", "at least one", and "one or more than one". In addition, the terms "having", "including", "containing", and "comprising" are interchangeable, and those skilled in the art recognize that these terms are open-ended terms.

[0101] The term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, according to the practice in the art, "about" may mean within 3 or more than 3 standard deviations. Alternatively, "about" may mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and still more preferably up to 1% above or below a given value. Alternatively, particularly with respect to biological systems or processes, the term may mean within an order of magnitude, preferably within 5 times, more preferably within 2 times of a value.

[0102] The term "alkyl" is known in the art and includes saturated aliphatic groups, including straight chain alkyl, branched chain alkyl, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl and cycloalkyl-substituted alkyl. In addition, the term "alkyl" (or "low alkyl") includes both "unsubstituted alkyl" and "substituted alkyl", wherein the substituted alkyl refers to an alkyl moiety in which a substituent replaces hydrogen on one or more carbons of the hydrocarbon backbone. These substituents may include, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thioformic acid / ester salt), alkoxy, phosphoryl, phosphonate / ester, phosphinate / ester, amino, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate / ester, sulfonate / ester, sulfamoyl, sulfonamido, sulfonyl, heterocyclic, aralkyl, or aromatic or heteroaromatic moieties. Those skilled in the art will appreciate that, if appropriate, the part itself substituted on the hydrocarbon chain may also be substituted. For example, the substituent of substituted alkyl may include substituted and unsubstituted forms of amino, azido, imino, amido, phosphoryl (including phosphonate / ester and phosphinate / ester), sulfonyl (including sulfate / ester, sulfonamido, sulfamoyl and sulfonate / ester), and silyl, as well as ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate / ester and ester),-CF ,-CN etc. Exemplary substituted alkyl is as described below. Cycloalkyl may further be substituted by alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl alkyl,-CF ,-CN etc.

[0103] The terms "amine" and "amino" are known in the art and include unsubstituted and substituted amines. Primary amines carry two hydrogens, secondary amines carry one hydrogen and one substituent, and tertiary amines have two hydrogens substituted. The substituents replacing one or two hydrogens may be, for example, alkyl, alkenyl, aryl, cycloalkyl, cycloalkenyl, heterocyclic, polycyclic, etc. If both hydrogens are replaced by carbonyls, the nitrogen surrounding the carbonyls forms an imide.

[0104] The term "alkylamine" embraces an amine group as defined above to which is appended a substituted or unsubstituted alkyl group.

[0105] The term "aryl" is known in the art and includes 5-, 6- and 7-membered monocyclic aromatic groups which may include 0 to 4 heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, etc. Those aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles" or "heteroaromatic compounds". The aromatic ring may be substituted at one or more ring positions by substituents as described above, such as halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphite, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclic, aromatic or heteroaromatic moiety, -CF3, -CN, etc. The term "aryl" also includes polycyclic ring systems having two or more rings in which two or more carbons are shared with two adjacent rings (the rings are "fused rings"), wherein at least one ring is aromatic, e.g., the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and / or heterocyclyl, or rings connected by non-cyclic moieties.

[0106] The terms "heterocyclyl" and "heterocyclic group" are art-recognized and include 3 to about 10 membered ring structures, such as 3 to about 7 membered rings, whose ring structures include 1 to 4 heteroatoms. The heterocycle may also be polycyclic. Heterocyclic groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxanthin, pyrrolidinazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, 2,3-diazine, 1,5-diazine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, dioxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as β-propiolactams (azetidinones), as well as pyrrolidones, sultams, sultones, and the like. The heterocycle may be substituted at one or more positions with substituents as described above, for example, halogen, alkylaralkyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphite, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic, aromatic or heteroaromatic moiety, -CD3, -CN, and the like.

[0107] The terms "polycyclic radical" and "polycyclic group" are known in the art and include structures having two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and / or heterocyclic radicals), wherein two or more carbons are common to two adjacent rings, e.g., the rings are "fused rings". Rings connected by non-adjacent atoms, e.g., three or more atoms are common to two rings, are referred to as "bridged" rings. Each of the polycyclic rings may be substituted with the above-mentioned substituents, e.g., halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphite, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic radical, aromatic or heteroaromatic moiety, -CD3, -CN, etc.

[0108] The term "carbocycle" is known in the art and includes aromatic or non-aromatic rings in which each atom of the ring is carbon. The following terms known in the art have the following meanings: "nitro" means -NO2; the term "halogen" means -F, -Cl, -Br, or -I; the term "sulfhydryl" means -SH; the term "hydroxyl" or "hydroxyl" means -OH; and the term "sulfonyl" means -SO2-.

[0109] The terms "amine" and "amino" are known in the art and include both unsubstituted and substituted amines. Primary amines carry two hydrogens, secondary amines carry one hydrogen and another substituent, and tertiary amines have two hydrogens substituted. The substituents for one or both hydrogens may be, for example, alkyl, alkenyl, aryl, cycloalkyl, cycloalkenyl, heterocyclic, polycyclic, etc. If two hydrogens are replaced by carbonyls, the nitrogen surrounding the carbonyls forms an imide.

[0110] The term "alkylamine" embraces an amine group as defined above to which is appended a substituted or unsubstituted alkyl group.

[0111] The term "amido" is art-recognized as referring to a carbonyl group substituted with an amino group.

[0112] The term "alkylthio" is known in the art and includes an alkyl group as defined above, to which a thio group is attached. In certain embodiments, the "alkylthio" moiety is represented by one of -S-alkyl, -S-alkenyl, -S-alkynyl, etc.

[0113] The term "carbonyl" is art-recognized and includes a C=O structure. The following are related to carbonyl groups: ester; carboxyl; formate; thiocarbonyl; thioester; thiocarboxylic acid; thioformate; ketone; and aldehyde.

[0114] The term "alkoxyl" or "alkoxy" is art-recognized and includes an alkyl group as defined above to which is attached an oxy group. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0115] An "ether" is two hydrocarbons covalently linked by an oxygen. Thus, a substituent of the alkyl group that makes the alkyl group an ether is or resembles an alkoxy group, which substituent may be represented, for example, by one of -O-alkyl, -O-alkenyl, -O-alkynyl, and the like.

[0116] The term "sulfonate" is art-recognized and includes moieties wherein the sulfur atom carries two doubly bonded oxygens and one singly bonded oxygen.

[0117] The term "sulfate" is art-recognized and includes a moiety that is similar to sulfonate but includes two singly bonded oxygens.

[0118] The terms "sulfonamide," "sulfamoyl," "sulfonyl," and "sulfoxido" are art-recognized, and each can include a variety of R group substituents as described herein.

[0119] The terms "phosphoramidite" and "phosphoramide" are known in the art.

[0120] The term "alkylselenyl" is known in the art and includes an alkyl group to which a substituted seleno group is attached. Exemplary "selenoethers" that may be substituted on the alkyl group are selected from one of -Se-alkyl, -Se-alkenyl, -Se-alkynyl, and the like.

[0121] Substitution may be made on alkenyl and alkynyl groups to produce, for example, aminoalkenyl, aminoalkynyl, amidoalkenyl, iminoalkenyl, iminoalkynyl, thioalkenyl, thioalkynyl, carbonyl substituted alkenyl or alkynyl groups.

[0122] It will be understood that "substitution" or "substituted with / by" includes the implicit precondition that such substitution complies with the allowed valencies of the substituted atom and the substituent and that the substitution results in a stable compound, e.g., the compound does not spontaneously transform, e.g., by rearrangement, cyclization, elimination or other reaction.

[0123] The term "substituted" also contemplates all allowed substituents of organic compounds (e.g., target imide reagents). In a broad sense, allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein. For appropriate organic compounds, allowed substituents may be one or more, which may be the same or different. For purposes of the present invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any allowed substituents of organic compounds described herein that satisfy the heteroatom valence state. The present invention is not intended to be limited in any way by the allowed substituents of organic compounds.

[0124] As used herein, the term "enantiomers" refers to a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of an enantiomeric pair is a "racemic" mixture. The term is used to refer to a racemic mixture where appropriate.

[0125] As used herein, the term "diastereomer" refers to a stereoisomer having at least two asymmetric atoms but not being mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. When a compound is a pure enantiomer, the stereochemistry of each chiral carbon can be specified with R or S. Resolved compounds with unknown absolute configuration can be specified as (+) or (-), depending on the direction (right-handed or left-handed) in which they rotate plane polarized light at the wavelength of the sodium D line. The compounds of the disclosed subject matter contain one or more asymmetric centers, and thus enantiomers, diastereomers, and other stereoisomeric forms can be produced, which can be defined as (R)- or (S)- according to absolute stereochemistry. The disclosed subject matter is intended to include all these possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents, or by splitting using conventional techniques. If the compound contains a double bond, the substituent can be in E or Z configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis or trans configuration. All tautomeric forms are also intended to be included.

[0126] As used herein, the term "isomer" refers to different compounds with the same molecular formula but different atomic arrangements and configurations. Also as used herein, the term "stereoisomer" refers to any of the various stereoisomer configurations that may exist for a given compound of the disclosed subject matter, and includes geometric isomers. It should be understood that substituents may be attached to the chiral center of a carbon atom. Therefore, the disclosed subject matter includes enantiomers, diastereomers, or racemates of a compound. In addition, as used herein, the term "constitutional isomer" refers to different compounds with the same number and type of atoms but atoms connected in different ways.

[0127] The term "carrier / vehicle" refers to a diluent, adjuvant, excipient or vehicle with which the therapeutic agent is administered. The physiological carrier may be a sterile liquid, such as water and oil, including liquids of petroleum, animal, plant or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. When the pharmaceutical composition is administered intravenously, water is a suitable carrier. Saline solutions as well as aqueous dextrose solutions and glycerol solutions may also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skimmed milk powder, glycerol, propylene glycol, water, ethanol, etc. If desired, the composition may also contain a small amount of a wetting agent or emulsifier, or a pH buffer.

[0128] For purposes of clarity and not by way of limitation, the detailed description of the disclosed subject matter is divided into the following subsections:

[0129] 5.1Tlr4 inhibitory compounds;

[0130] 5.2 Pharmaceutical compositions;

[0131] 5.3 Disorders / diseases; and

[0132] 5.4 Treatment methods

[0133] 5.1 TLR4 inhibitory compounds

[0134] The present disclosure provides carbon analogs of pyranose derivatives. In certain embodiments, the compounds can be used to inhibit Tlr4.

[0135] In certain embodiments, the present disclosure provides compounds of Formula I:

[0136]

[0137] wherein X is O, Z is O or NH, Y is CH2 or CH(R1), R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2, or

[0138] wherein X is CH2, Z is O or NH, and Y is O, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

[0139] In certain embodiments, the present disclosure provides compounds of Formula II:

[0140]

[0141] wherein Z is O or NH, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

[0142] In certain embodiments, the compounds of the present disclosure are shown in Table 1.

[0143] Table 1.

[0144]

[0145]

[0146]

[0147]

[0148]

[0149] In certain embodiments, a compound is considered to inhibit Tlr4 if it inhibits one or more signs or symptoms of inflammation, such as activation of nuclear factor kappa light-chain-enhancer of activated B cells ("NFkB"), increased expression / levels of interleukins (including but not limited to interleukin 1 ("Il1") and interleukin 6 ("Il6"), increased expression / levels of tumor necrosis factor ("Tnf"), increased expression / levels of lipocalin-2 ("Lcn2"), increased erythrocyte sedimentation rate, increased C-reactive protein, fever, shortness of breath, lethargy, swelling, redness, and / or pain. In certain embodiments, the ability of a compound and / or a particular concentration of a compound to inhibit Tlr4 can be determined using a Tlr4 activity assay, which can assess one of the signs or symptoms listed above. For example, in certain embodiments, Tlr4 inhibition can be determined using methods that measure the effect of a compound on NFkB activity, such as, but not limited to, a NFkB luciferase reporter mouse model stimulated with a Tlr4 ligand (e.g., LPS), as described in the Examples below, or HEK-Blue-4 cells (InvivoGen). Certain other non-limiting examples of systems for testing compounds to determine Tlr4 inhibitory activity include: CWT mice can be treated with LPS and a test compound and monitored for signs and symptoms of inflammation, and / or C3H / WT cells (InvivoGen) can be treated with LPS and a test compound and tested for NFkB activation, Il6 production, or other markers of inflammatory processes.

[0150] 5.2 Pharmaceutical composition

[0151] In certain embodiments, the present disclosure provides a pharmaceutical composition comprising a compound of Formula I or Formula II as described above in a suitable pharmaceutical carrier. When administered to a subject in need of such treatment, the amount of the compound present in the composition can be calculated so as to provide an effective amount of a compound of Formula I or Formula II.

[0152] In certain embodiments, the disclosure provides pharmaceutical compositions comprising a therapeutically effective amount of any of the compounds of Formula I or Formula II, for example but not limited to, together with a pharmaceutical carrier such as water or other physiological solvents. The therapeutically effective amount inhibits Tlr4.

[0153] In non-limiting embodiments, the compound of Formula I or Formula II may be included in coated particles, micelles, liposomes, or similar structures.

[0154] In certain embodiments, the pharmaceutical composition may be a liquid, including a compound of Formula I or Formula II in a liquid drug carrier, such as water (aqueous carrier) or saline. In certain embodiments, the liquid composition may optionally further contain one or more of a buffer or a preservative.

[0155] In certain other embodiments, the pharmaceutical composition of the present disclosure may be solid, for example in the form of tablets, capsules, sachets or suppositories, including a dose of a compound of Formula I or Formula II, which, when administered according to a dosing regimen, provides an effective amount of a compound of Formula I or Formula II for an object in need of the treatment. In certain embodiments, the solid pharmaceutical composition may further include one or more excipients, such as, but not limited to, lactose, sucrose, mannitol, erythritol, carboxymethyl cellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, starch, polyvinyl pyrrolidone, and the like.

[0156] In certain embodiments, the pharmaceutical composition may include an additive with antimicrobial and / or anti-inflammatory activity. In certain embodiments, the compound includes, but is not limited to, an antibiotic, a steroid, or a non-steroidal anti-inflammatory agent. In certain other embodiments, the pharmaceutical composition may include an analgesic. In certain further embodiments, the pharmaceutical composition may include an agent that improves cardiac function and / or reduces cardiac stress, such as, but not limited to, angiotensin converting enzyme inhibitors, beta blockers, nitroglycerin or related nitrate / ester compounds, digoxin or related compounds, or calcium channel blockers.

[0157] Pharmaceutically acceptable salts are known in the art and include relatively nontoxic inorganic and organic acid addition salts of the compositions of the present invention, including, but not limited to, therapeutic agents, excipients, other materials, and the like. Examples of pharmaceutically acceptable salts include salts derived from inorganic acids, such as hydrochloric acid and sulfuric acid, and salts derived from organic acids, such as ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. Examples of suitable inorganic bases for forming salts include hydroxides, carbonates, and bicarbonates of ammonia, sodium, lithium, potassium, calcium, magnesium, aluminum, zinc, and the like. Salts may also be formed with suitable organic bases, including those that are nontoxic and strong enough to form such salts. For illustrative purposes, the class of organic bases may include monoalkyl, dialkyl and trialkylamines, such as methylamine, dimethylamine and triethylamine; monohydroxyalkyl, dihydroxyalkyl or trihydroxyalkylamines, such as monoethanolamine, diethanolamine and triethanolamine; amino acids, such as arginine and lysine; guanidine; N-methylglucosamine; N-methylglucosamine; L-glutamine; N-methylpiperazine; morpholine; ethylenediamine; N-benzylphenethylamine; (trihydroxymethyl)aminoethane; etc., for example, see J. Pharm. Sci., 66: 1-19 (1977).

[0158] 5.3 Disorders / diseases

[0159] In certain embodiments, the compounds, compositions and methods provided herein can be used to treat any disease / disorder ("disorder") involving Tlr44 activation, including but not limited to infectious diseases and inflammatory disorders, such as sepsis, necrotizing enterocolitis ("NEC"), autoimmune diseases, Crohn's disease, celiac disease, ulcerative colitis, rheumatoid arthritis, cardiovascular disease (including myocardial infarction), epilepsy, gram-negative bacterial infections, aspergillosis, periodontal disease, Alzheimer's disease, cigarette smoke-mediated lung inflammation, viral hepatitis (including hepatitis C virus hepatitis), alcoholic hepatitis, insulin resistance in adipocytes, osteoarthritis, pancreatitis, metabolic syndrome, trauma-induced systemic inflammation, acute respiratory distress syndrome (ARDS), COVID-19-induced systemic inflammation, organ rejection after transplantation, etc. See, e.g., U.S. Patent Application Publication No. 2008 / 0311112A1, published on December 18, 2008. In non-limiting embodiments, the presently disclosed subject matter can also be used to treat post-traumatic conditions, including ischemic and traumatic injuries to the heart, liver, lung, kidney, intestine, brain, eye, and pancreas.

[0160] 5.4 Treatment

[0161] In certain embodiments, the disclosure provides methods of treating an infectious or inflammatory disorder, comprising administering to a subject in need of such treatment an effective amount of a compound of Formula I or Formula II that reduces one or more signs or symptoms of the subject.

[0162] In certain embodiments, the disclosure provides a method for treating an intestinal inflammatory disorder in a subject, the method comprising administering to a subject in need of such treatment an effective amount of a compound of Formula I or Formula II that reduces intestinal inflammation in the subject. In certain specific embodiments, the intestinal inflammatory disorder is necrotizing enterocolitis. In certain embodiments, the disclosure provides a method for treating and / or preventing necrotizing enterocolitis in premature infants.

[0163] In certain embodiments, the disclosure provides methods of treating inflammatory lung disease in a subject, the methods comprising administering to a subject in need of such treatment an effective amount of a compound of Formula I or Formula II that reduces pulmonary airway inflammation in the subject.

[0164] In certain embodiments, the present disclosure provides a method for treating traumatic injury in a subject, the method comprising administering to a subject in need of such treatment an effective amount of a compound of Formula I or Formula II that reduces Tlr4-induced post-traumatic injury. In certain specific embodiments, the traumatic injury is directed to an organ selected from the group consisting of the heart, liver, lung, kidney, intestine, brain, eye, and pancreas.

[0165] In certain embodiments, the present disclosure provides methods of treating one or more of the following: osteoarthritis, pancreatitis, metabolic syndrome, trauma-induced systemic inflammation, acute respiratory distress syndrome (ARDS), COVID-19-induced systemic inflammation, organ rejection after transplantation, or other gastrointestinal disorders that may involve Tlr44 signaling, such as ulcerative colitis or Crohn's disease.

[0166] In certain embodiments, the subject is a human or non-human subject. In certain embodiments, the subject is a human subject. In certain embodiments, the subject is a non-human subject.

[0167] In certain embodiments, the compound of Formula I or Formula II can be administered by any standard route, including but not limited to: oral, intraperitoneal (ip), intravenous (iv), subcutaneous (sc), intradermal, intramuscular (im), intraarticular, intrathecal, intraarterial, intravaginal, rectal, nasal, and pulmonary.

[0168] In certain embodiments, the effective dose may be determined using methods known in the art, including but not limited to the Tlr4 activity assay described herein. In certain embodiments, the effective dose may be between about 0.01 micromoles and about 50 micromoles of the compound of Formula I or Formula II per kilogram of the subject's weight, or between about 0.1 micromoles and about 20 micromoles of the compound of Formula I or Formula II per kilogram of the subject's weight.

[0169] In certain embodiments, the dosage of the compound of Formula I or Formula II can be between about 0.001 mg and about 100 mg per kilogram of the subject's weight, between about 0.01 mg and about 10 mg per kilogram of the subject's weight, between about 0.1 mg and about 10 mg per kilogram of the subject's weight, or between about 0.5 and 5 mg per kilogram of the subject's weight.

[0170] 6. Example

[0171] The presently disclosed subject matter may be better understood by reference to the following examples, which are provided by way of illustration and not by way of limitation.

[0172] Example 1: Exemplary Synthesis of Carbon Analogues of Pyranose Derivatives

[0173] This example provides the synthesis of carbon analogs of pyranose derivatives according to certain embodiments of the present disclosure.

[0174] All reactions were performed under an N2 atmosphere passed through a Drierite column. All glassware was flame dried under high vacuum or dried in an oven overnight and cooled under a N2 flow before use. Reactions were magnetically stirred using a Teflon-coated magnetic stir bar, and syringe needles were dried in an oven and cooled over Drierite in a dry cabinet. CH2Cl2 was freshly distilled over calcium hydride. All other materials were from commercial sources and used as received. Reactions were monitored by thin layer chromatography (TLC) on pre-coated silica gel 60 F254 plates (250 μm layer thickness); visualization was achieved by UV light (254 nm) and / or staining with KMnO4 solution (1.5 g KMnO4 and 10 g K2CO3 in 200 mL H2O with 10% NaOH) or PMA solution (10 g phosphomolybdic acid in 100 mL anhydrous ethanol). Flash chromatography was performed on silica gel 60 (230–400 mesh). Melting points were determined in open capillaries and recorded on a Mel-Temp II instrument equipped with a Fluke 51II digital thermometer and are uncorrected. Nuclear magnetic resonance (NMR) spectra were collected on a Bruker instrument, 1 H and 13C was operated at 300, 400, 500 and 600 MHz at ambient temperature. Chemical shifts (δ) are reported in parts per million (ppm) and the residual solvent peak was used as an internal standard (CDCl3: 1 H NMR 7.26 ppm, 13 C NMR 77.16 ppm). 1 H NMR is tabulated as follows: chemical shift, multiplicity (s = singlet, d = doublet, t = triplet, m = multiplet, br = broad), coupling constant, and number of protons. High-resolution mass spectra (HRMS) were obtained on a Thermo Scientific Exactive Orbitrap LC-MS coupled to a Thermo Scientific Accela HPLC system (ESI positive ion mode using a 3.5 μM Water XTerra C18 column (2.1 × 50 mm, 10 min gradient elution with MeCN / H2O / MeOH containing 0.1% formic acid, flow rate 500 μL / min, from 3:92:5 at 0–0.5 min to 93:2:5 at 4.0 min, back to 3:92:5 at 6.0–7.5 min).

[0175] This example provides the following syntheses: (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-6-propyltetrahydro-2H-pyran-3,4,5-triyl triacetate (7), (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-6-isobutyltetrahydro-2H-pyran-3,4,5-triyl triacetate, (8) (2R,3S,4R,5R,6R)-2-isobutyl-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4 ,5-triyl tris(2,2-dimethylpropionate), (9) (2R,3R,4R,5S,6R)-2-(((3,3-dimethylbutanoyl)oxy)methyl)-6-isobutyltetrahydro-2H-pyran-3,4,5-triyl tris(3,3-dimethylbutanoate) (10), and (2R,3R,4R,5S,6R)-2-((benzoyloxy)methyl)-6-isobutyltetrahydro-2H-pyran-3,4,5-triyl tribenzoate (11), as shown in Scheme I below. Intermediates 3 and 4 were disclosed in procedures previously reported in the literature.

[0176]

[0177] Example 1.1 - Synthesis of (2R,3S,4R,5R,6R)-2-(Hydroxymethyl)-6-propyltetrahydro-2H-pyran-3,4,5-triol (5):

[0178] A suspension of 3 and 20% Pd(OH) / C in absolute ethanol was stirred at room temperature under H (balloon). The reaction mixture was filtered through celite, the filter pad was rinsed with EtOH, and the filtrate was concentrated under reduced pressure and dried under vacuum to give the crude product 5, which was used in the subsequent step without further purification.

[0179] Example 1.2 - Synthesis of (2R,3S,4R,5R,6R)-2-(Hydroxymethyl)-6-isobutyltetrahydro-2H-pyran-3,4,5-triol (6):

[0180] A suspension of 4 (550 mg, 0.950 mmol) and 20% Pd(OH)2 / C (123 mg) in absolute ethanol (12 mL) was stirred under H2 (balloon) at room temperature for 16 h. The reaction mixture was filtered through celite, the filter pad was rinsed with EtOH, and the filtrate was concentrated under reduced pressure and dried under vacuum to give 209 mg of crude tetraol (100%, 0.96 mmol) as a colorless solid, which was used in the subsequent step without further purification. 1 H NMR(MeOD,500MHz)δ4.02-3.98(m,1H),3.76(dd,J=11.7,2.3Hz,1H),3.59(dd,J=11.7,1.0Hz,1H),3.51(t,J=8.7Hz,1H),3.42-3.38(m,1H),3.2 5(t,J=8.7Hz,1H),1.82-1.78(m,1H),1.72-1.66(m,1H),1.38-1.33(m, 1H), 1.18 (t, J = 7.0Hz, 1H), 0.97 (d, J = 6.7Hz, 3H), 0.93 (d, J = 6.5Hz, 3H).

[0181] Example 1.3 - Synthesis of (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-6-propyltetrahydro-2H-pyran-3,4,5-triyl triacetate (7):

[0182] To a suspension of 5 (100 mg, 0.49 mmol) and acetic anhydride (0.23 mL, 2.4 mmol) in pyridine (0.2 mL) was added a catalytic amount of DMAP. The solution was stirred at room temperature for 16 hours and diluted with ethyl acetate (10 mL). The organic phase was washed with 1M HCl (3 x 5 mL), water (1 x 5 mL), saturated aqueous NaHCO3 (1 x 5 mL) and brine (5 mL x 1) in sequence. The organic phase was dried (Na2SO4), concentrated, and the residue was dried under vacuum to give α and β pentaacetylated end isomers in a ratio of 4:1. Recrystallization from a hexane / EtOAc mixture gave 137 mg of 7 (75%, 0.366 mmol) as white needles. Melting point 102–103.5°C. 1 H NMR(CDCl3,500MHz)δ5.32(t,J=9.3Hz,1H),5.07(dd,J=9.3,5.8Hz,1H),4.99(t, J=9.3Hz,1H),4.24(dd,J=12.2,5.3Hz,1H),4.20-4.16(m,1H),4.08(dd,J=12.2, 2.5Hz,1H),3.83-3.80(m,1H),2.09(s,3H),2.05(s,3H),2.03(s,3H),2.03(s,3H ),1.81-1.74(m,1H),1.50-1.41(m,2H),1.35-1.29(m,1H),0.97(t,J=7.1Hz,3H). 13 C NMR (CDCl3, 500 MHz) δ 170.8, 170.4, 169.8, 169.7, 72.6, 70.7, 69.1, 68.7, 62.5, 27.5, 20.9, 20.8, 18.4, 13.9. HRMS calculated C 17 H 27 O9[M+H] + :375.1650; Found:375.1647.

[0183] Example 1.4 - Synthesis of (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-6-isobutyltetrahydro-2H-pyran-3,4,5-triyl triacetate (8):

[0184] To a solution of 6 (42 mg, 0.19 mmol) and a catalytic amount of DMAP in pyridine (0.65 mL) was added acetic anhydride (0.18 mL, 1.91 mmol). The reaction mixture was stirred at room temperature for 16 hours and diluted with EtOAc (5 mL). The organic phase was washed successively with 1M HCl (5 mL x 2), saturated NaHCO3 aqueous solution (5 mL x 2) and brine (5 mL x 1). The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified on SiO2 (hexane / EtOAc, 8:1) to give 50 mg of 8 (76%, 0.129 mmol) as an amorphous solid. Melting point 58.5-60°C. 1 H NMR(CDCl3,400MHz)δ5.31(t,J=9.2Hz,1H),5.06(dd,J=9.6,5.8Hz,1H),4.99(t,J =9.2Hz,1H),4.30-4.24(m,1H),4.24(dd,J=12.1,5.1Hz,1H),4.06(dd,J=12.1,2. 6Hz,1H),3.84-3.80(m,1H),2.08(s,3H),2.05(s,3H),2.03(s,3H),2.02(s,3H),1 .82-1.78(m,2H),1.27-1.18(m,1H),0.97(d,J=6.6Hz,3H),0.91(d,J=6.5Hz,3H). 13 C NMR (CDCl3, 400 MHz) δ 170.8, 170.4, 169.8, 169.7, 71.0, 70.7, 70.6, 69.1, 68.8, 62.5, 33.8, 24.2, 23.6, 21.5, 20.9. HRMS calculated for C 18 H 29 O9[M+H] + :389.1806; Found 389.1801.

[0185] Example 1.5 - Synthesis of (2R,3S,4R,5R,6R)-2-isobutyl-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyl tris(2,2-dimethylpropionate) (9):

[0186] To a solution of 6 (50 mg, 0.23 mmol) and a catalytic amount of DMAP in pyridine (0.8 mL) was added pivaloyl chloride (0.28 mL, 2.23 mmol). The reaction mixture was stirred at room temperature for 16 hours and diluted with EtOAc (5 mL). The organic phase was washed successively with 1M HCl (5 mL x 2), saturated NaHCO3 aqueous solution (5 mL x 2) and brine (5 mL x 1). The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified on SiO2 (hexane / EtOAc, 18:1) to obtain 81 mg of 9 (64%, 0.146 mmol) as an amorphous solid. Melting point 123-124 ° C. 1 H NMR(CDCl3,600MHz)δ5.41(t,J=9.7Hz,1H),5.06(dd,J=10.0Hz,1H),5.01(t,J=9 .7Hz,1H),4.29-4.25(m,1H),4.12(dd,J=12.0,1.2Hz,1H),4.00(dd,J=12.0,6.4 Hz,1H),3.82-3.80(m,1H),1.88-1.83(m,1H),1.76-1.71(m,1H),1.21(s,9H),1. 17(s,9H),1.16(s,9H),1.12(s,9H),0.98(d,J=6.7Hz,3H),0.88(d,J=6.5Hz,3H). 13 C NMR (CDCl3, 600 MHz) δ 178.3, 177.3, 177.2, 176.8, 71.0, 70.9, 70.2, 68.9, 63.0, 39.0, 38.9 (2 signals), 33.4, 27.3 (2 signals), 27.2, 23.8 (2 signals), 21.3. HRMS calculated for C 30 H 53 O9[M+H] + :557.3684; Found:557.3680.

[0187] Example 1.6 - Synthesis of (2R,3R,4R,5S,6R)-2-(((3,3-dimethylbutanoyl)oxy)methyl)-6-isobutyltetrahydro-2H-pyran-3,4,5-triyl tris(3,3-dimethylbutyrate) (10):

[0188] 3,3-Dimethylbutyryl chloride (0.32 mL, 2.27 mmol) was added to a solution of 6 (50 mg, 0.23 mmol) and a catalytic amount of DMAP in pyridine (0.8 mL). The reaction mixture was stirred at room temperature for 16 hours and diluted with EtOAc (5 mL). The organic phase was washed with 1M HCl (5 mL x 2), saturated NaHCO3 aqueous solution (5 mL x 2) and brine (5 mL x 1) in sequence. The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified on SiO2 (hexane / EtOAc, 20:1) to give 118 mg of 10 (85%, 0.146 mmol) as an amorphous solid. Melting point 83-84 ° C. 1 H NMR(CDCl3,500MHz)δ5.37(t,J=9.3Hz,1H),5.03(dd,J=9.7Hz,1H),4.98(t,J=9.1Hz,1 H),4.29-4.25(m,1H),4.15-4.09(m,2H),3.84-3.80(m,1H),2.23(s,2H),2.22-2.11(m ,4H),2.15(s,2H),1.82-1.75(m,1H),1.74-1.69(m,1H),1.24-1.19(m,1H),1.02(s,9H ), 1.01 (s, 9H), 1.00 (s, 9H), 0.98 (s, 1H), 0.95 (d, J = 6.7Hz, 3H), 0.88 (d, J = 6.5Hz, 3H). 13 C NMR (CDCl3, 500 MHz) δ 172.0, 171.3, 171.1, 170.9, 70.9, 70.7, 69.8, 69.2, 69.0, 62.5, 47.8 (2 signals), 47.7, 47.6, 33.9, 30.9, 30.8, 30.7, 30.6, 29.7 (3 signals), 24.1, 23.7, 21.5. HRMS calculated for C 34 H 61 O9[M+H] + :613.4310; Found:613.4307.

[0189] Example 1.7 - Synthesis of (2R,3R,4R,5S,6R)-2-((Benzoyloxy)methyl)-6-isobutyltetrahydro-2H-pyran-3,4,5-triyl tribenzoate (11):

[0190] Benzoyl chloride (0.18 mL, 1.54 mmol) was added to a solution of 6 (34 mg, 0.154 mmol) and a catalytic amount of DMAP in pyridine (0.8 mL). The reaction mixture was stirred at room temperature for 16 hours and diluted with EtOAc (5 mL). The organic phase was washed with 1M HCl (5 mL x 2), saturated NaHCO3 aqueous solution (5 mL x 2) and brine (5 mL x 1) in sequence. The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified on SiO2 (hexane / EtOAc, 18:1) to give 73 mg of 11 (82%, 0.127 mmol) as a white crystalline solid. Melting point 129.5-131 ° C. 1 H NMR(CDCl3,500MHz)δ8.04(d,J=7.2Hz,2H),7.98(d,J=7.3Hz,2H),7.93(d,J=7.3Hz,2H),7.90( d,J=7.3Hz,2H),7.57-7.38(m,8H),7.35-7.31(m,4H),5.99(t,J=9.0Hz,1H),5.57(t,J=9.0Hz, 1H),5.50(dd,J=9.0,5.7Hz,1H),4.61(m,1H),4.55(d,J=4.7Hz,1H),4.31-4.28(m,1H),2.09-2 .03(m,1H),1.87-1.81(m,1H),1.40-1.35(m,1H),0.99(d,J=6.7Hz,3H),0.90(d,J=6.5Hz,3H). 13 C NMR (CDCl3, 500 MHz) δ 166.4, 166.0, 165.5 (2 signals), 133.6, 133.5, 133.4, 133.2, 130.0 (2 signals), 129.9, 129.3, 129.2, 129.1, 128.6, 71.6, 71.0, 70.7, 70.0, 69.7, 63.5, 34.4, 24.3, 23.7, 21.5. HRMS calculated for C 38 H 37 O9[M+NH4] + :654.2698; Found:654.2684.

[0191] This example further provides the synthesis of (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-6-isobutyl-5-(pivaloyloxy)tetrahydro-2H-pyran-3,4-diyl acetate (14), and (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-5-((3,3-dimethylbutanoyl)oxy)-6-isobutyltetrahydro-2H-pyran-3,4-diyl diacetate (15), as shown in Scheme II.

[0192]

[0193] Example 1.8 - Synthesis of (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-5-(benzyloxy)-6-isobutyltetrahydro-2H-pyran-3,4-diyl diacetate (12):

[0194] The suspension of 4 (787mg, 1.36mmol) and 20% Pd (OH) / C (176mg) in EtOH (17mL) was stirred at room temperature for 16 hours under H (balloon). The reaction mixture was filtered through diatomite, and the filter pad was rinsed with ethanol, and the filtrate was concentrated and dried under vacuum to obtain 348mg of a 2:1 mixture of tetraol and triol as a colorless solid. Acetic anhydride (1.3mL, 13.6mmol) was added dropwise to a solution of crude intermediate and catalytic amount of DMAP. The reaction mixture was stirred at room temperature for 16 hours and diluted with EtOAc (10mL). The organic phase was washed successively with 1M HCl (10mL x 2), saturated NaHCO aqueous solution (10mL x2) and salt solution (10mL x 2mL). The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified on SiO2 (hexane / EtOAc, 4:1) to afford 165 mg of 12 (28%, 0.378 mmol) as a white crystalline solid. 1 H NMR(CDCl3,500MHz)δ7.25-7.26(m,4H),5.28(t,J=9.5Hz,1H),4.91(t,J=9.5Hz,1H) ,4.56(d,J=3.9Hz,2H),4.25(dd,J=12.2,4.9Hz,1H),4.12-4.09(m,1H),4.00(dd,J=1 2.2,2.3Hz,1H),3.80-3.77(m,1H),3.68(dd,J=9.7,5.9Hz),2.06(s,3H),2.01(s,6H ),1.80-1.65(m,2H),1.43-1.40(m,1H),0.96(d,J=6.7Hz,3H),0.89(d,J=6.5Hz,3H). 13C NMR(CDCl3,500MHz)δ170.9,170.5,170.0,137.9,128.6,128.1,127.9,73 .2,72.5,72.4,69.5,68.4,62.7,33.0,24.3,23.7,21.5,21.0,20.9,20.8.

[0195] Example 1.9 - Synthesis of (2R,3R,4R,5S,6R)-2-(acetoxymethyl)-5-hydroxy-6-isobutyltetrahydro-2H-pyran-3,4-diyl diacetate (13):

[0196] A solution of 12 (140 mg, mmol) and 20% Pd(OH)2 / C (31 mg) in EtOH (4 mL) was stirred under H2 (balloon) at room temperature for 16 h. The reaction mixture was passed through celite, the filter pad was rinsed with ethanol, and the filtrate was concentrated under reduced pressure to give 100 mg of 13 (90%, 0.29 mmol) as a colorless solid. Melting point 73-76°C. 1 H NMR(CDCl3,400MHz)δ5.10(t,J=8.4Hz,1H),4.94(t,J=8.4Hz,1H),4.31(dd, J=12.1,5.6Hz,1H),4.17-4.12(m,1H),4.07(dd,J=12.1,5.6Hz,1H),389-3. 83(m,1H),2.23(d,J=6.6Hz,1H),2.10(s,3H),2.08(s,3H),2.05(s,3H),1.7 7-1.68(m,2H),1.47-1.40(m,1H),0.99(d,J=6.5,3H),0.95(d,J=6.5Hz,3H). 13 C NMR (CDCl3, 500 MHz) δ 171.4, 170.9, 169.7, 73.7, 72.9, 70.2, 69.5, 68.7, 62.4, 33.7, 24.4, 23.7, 21.7, 21.1, 20.9 (2 signals). HRMS calculated for C 16 H 27 O8[M+H] + :347.1700; Found:347.1698.

[0197] Example 1.10 - (2R,3R,4R,5S,6R)-2-(Acetoxymethyl)-6-isobutyl-5-(pivaloyloxy)tetrahydro-2H-pyran-3,4-diyl diacetate (14):

[0198] At room temperature, to 13 (48mg, 0.14mmol), pyridine (0.1mL) and catalytic amount of DMAP in anhydrous CH2Cl2 (1.5mL) solution add pivaloyl chloride (0.042mL, 0.346mmol). The reaction mixture was stirred for 16 hours and diluted with CH2Cl2 (5mL). The organic phase was washed successively with 1M HCl aqueous solution (5mL x 2), saturated NaHCO3 aqueous solution (5mL x 2) and saline (5mL x1). The organic phase was dried (Na2SO4) and concentrated under reduced pressure. On SiO2 (hexane / EtOAc, 8:1) purification of the obtained residue, 19mg of 14 (32%, 0.044mmol) was obtained as a colorless amorphous solid. 1 H NMR(CDCl3,600MHz)δ5.35(t,J=9.3Hz,1H),5.02(dd,J=10.1Hz,1H),5.00(t,J=9 .3Hz,1H),4.30-4.28(m,1H),4.26(dd,J=12.2,5.2Hz,1H),4.06(dd,J=12.2,2.4 Hz,1H),3.83(m,1H),2.09(s,1H),2.03(s,1H),2.00(s,1H),1.82-1.77(m,1H),1 .73-1.68(m,1H),1.19-1.14(m,1H),0.97(d,J=6.7Hz,3H),0.90(d,J=6.5Hz,3H). 13 C NMR (CDCl3, 600 MHz) δ 177.2, 170.9, 170.3, 169.8, 71.3, 70.5, 70.4, 69.1, 68.8, 62.6, 38.9, 33.6, 27.1, 24.3, 23.7, 21.6, 20.9, 20.8. HRMS calculated for C 21 H 35 O9[M+H] + :431.2276; Found:431.2269.

[0199] Example 1.11 (2R, 3R, 4R, 5S, 6R)-2-(acetoxymethyl)-5-((3,3-dimethylbutanoyl)oxy)-6-isobutyltetrahydro-2H-pyran-3,4-diyl diacetate (15):

[0200] At room temperature, 3,3-dimethylbutyryl chloride (0.05 mL, 0.35 mmol) was added to a solution of 13 (48 mg, 0.14 mmol), pyridine (0.1 mL) and a catalytic amount of DMAP in anhydrous CH2Cl2 (1.5 mL). The reaction mixture was stirred for 16 hours and diluted with CH2Cl2 (5 mL). The organic phase was washed with 1M HCl aqueous solution (5 mL x 2), saturated NaHCO3 aqueous solution (5 mL x 2), and brine (5 mL x 1) in sequence. The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified on SiO2 (hexane / EtOAc, 9:1) to obtain 29 mg of 15 (47%, 0.065 mmol) as a colorless amorphous solid. 1 H NMR(CDCl3,600MHz)δ5.34(t,J=9.5Hz,1H),5.08(dd,J=9.5,5.9Hz,1H),4.98(t,J=9.5Hz ,1H),4.32-4.28(m,1H),4.24(dd,J=12.2,5.0Hz,1H),4.06(dd,J=12.2,2.3Hz,1H),3.84- 3.81(m,1H),2.19(s,2H),2.08(s,3H),2.03(s,3H),2.00(s,3H),1.81-1.76(m,1H),1.72- 1.68(m,1H),1.27-1.19(m,1H),1.00(s,9H),0.97(d,J=6.7Hz,3H),0.90(d,J=6.5Hz,3H). 13 CNMR (CDCl3, 600MHz)δ171.1,170.8,170.4,169.8,71.2,70.6,70.3,69.4,68.6,62.5,47.8,33.6,30.9,29.6,24.2,23.7,21.5,20.9 (2 signals),20.8. HRMS calculated C 22 H 37 O9[M+H] + :445.2432; Found:445.2425.

[0201] This example further provides a proposed synthesis of (2R,3S,4R,5S,6R)-2-(acetoxymethyl)-6-isobutyl-5-(pivaloyloxy)tetrahydro-2H-pyran-3,4-diyl diacetate (21), as shown in Scheme III.

[0202]

[0203]

[0204] Example 1.12 - Synthesis of (2R,3S,4R,5S,6R)-2-(acetoxymethyl)-5-(benzyloxy)-6-isobutyltetrahydro-2H-pyran-3,4-diyl diacetate (19):

[0205] A suspension of 18 and 20% Pd(OH)2 / C in EtOH was stirred at room temperature under H2 (balloon). The reaction mixture was passed through diatomaceous earth, the filter pad was rinsed with EtOH, and the filtrate was concentrated and dried under vacuum. Acetic anhydride was added dropwise to a solution of the crude intermediate and a catalytic amount of DMAP. The reaction mixture was stirred at room temperature and diluted with EtOAc. The organic phase was washed successively with 1M HCl, saturated NaHCO3 aqueous solution and brine. The organic phase was dried (Na2SO4) and concentrated under reduced pressure to give 19 (see, e.g., Cipolla, L.; Lay, L.; Nicotra, FJ Org. Chem. 1997, 62 (19), 6678-6681).

[0206] Example 1.13 - Synthesis of (2R,3S,4R,5S,6R)-2-(acetoxymethyl)-5-hydroxy-6-isobutyltetrahydro-2H-pyran-3,4-diyl diacetate (20):

[0207] A solution of 19 and 20% Pd(OH)2 / C in EtOH was stirred at room temperature under H2 (balloon). The reaction mixture was passed through celite, the filter pad was rinsed with EtOH, and the filtrate was concentrated under reduced pressure to give 20.

[0208] Example 1.14 - (2R,3S,4R,5S,6R)-2-(Acetoxymethyl)-6-isobutyl-5-(pivaloyloxy)tetrahydro-2H-pyran-3,4-diyl diacetate (21):

[0209] At room temperature, pivaloyl chloride was added to a solution of 20, pyridine, and a catalytic amount of DMAP in anhydrous CH2Cl2. The reaction mixture was stirred and then diluted with CH2Cl2. The organic phase was washed with 1M HCl aqueous solution, saturated NaHCO3 aqueous solution, and brine in sequence. The organic phase was dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified on SiO2 to give 21.

[0210] This example further provides a suggested synthesis of (2R,3S,4R,5R,6S)-5-acetylamino-2-(acetoxymethyl)-6-(prop-2-yn-1-yloxy)tetrahydro-2H-pyran-3,4-diyl diacetate (24), as described in Scheme IV.

[0211]

[0212] Example 1.15 - Synthesis of (2R,3S,4R,5R,6S)-5-acetylamino-2-(acetoxymethyl)-6-(prop-2-yn-1-yloxy)tetrahydro-2H-pyran-3,4-diyl diacetate (24):

[0213] A suspension of 22, acetic anhydride, and montmorillonite K-10 was stirred to give 23. Subsequently, a solution of 23, 5% HCl prepared in situ, and propargyl alcohol were stirred at 65°C. To this solution, acetic anhydride and pyridine were added. The solution was then stirred at room temperature to give 24 (see, e.g., Wipf, P.; Eyer, BR; Yamaguchi, Y.; Zhang, F.; Neal, MD; Sodhi, CP; Good, M.; Branca, M.; Prindle, T., Jr.; Lu, P.; Brodsky, JL; Hackam, DJ Tetrahedron Letters. 2015, 56(23), 3097-3100).

[0214] This example further provides the synthesis of (2R,3S,4R,5S,6R)-2-(acetoxymethyl)-5-pivaloylamino-6-propyltetrahydro-2H-pyran-3,4-diyl diacetate (28), as shown in Scheme V.

[0215]

[0216] Example 1.16 - N-((2R,3S,4R,5S,6R)-2-allyl-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)pivalamide (26):

[0217] To a solution of 25 (265 mg, 0.560 mmol) and a catalytic amount of DMAP in an 8:1 mixture of CH2Cl2 and pyridine (3 mL) was added pivaloyl chloride (0.082 mL, 0.67 mmol) in one portion at room temperature. After 6 h, the reaction mixture was concentrated under reduced pressure and the resulting residue was purified by chromatography on SiO2 (hexane / EtOAc, 4:1) to afford 26 (194 mg, 62%) as a colorless solid: mp 32-34°C; HRMS m / z calculated for C 35 H 44 O5N([M+1] + )558.3214, found 558.3213; 1H NMR(500MHz, CDCl3)δ7.35-7.20(m,15H),6.79(d,J=9.5Hz),5.83-5.78(m,1 H),5.10-5.02(m,2H),4.63-4.57(m,2H),4.52(s,2H),4.49-4.43(m,2H),4. 34-4.32(m,1H),3.95-3.93(m,1H),3.88(dd,J=7.0,10.0Hz),3.73(dd,J=7. 0,10.0Hz),3.65(bs,1H),3.58-3.57(m,1H),2.25-2.10(m,1H),1.04(s,9H).

[0218] Example 1.17 - N-((2R,3R,4R,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-propyltetrahydro-2H-pyran-3-yl)pivalamide (27):

[0219] In a Parr reactor, a slurry of 26 (134 mg, 1.14 mmol) and 20% Pd(OH)2 / C (30 mg) in EtOH (3.5 mL) was stirred under H2 (100 psi) at room temperature for 48 h. The reaction mixture was passed through The filtrate was concentrated under reduced pressure to give 27 (66 mg, 0.23 mmol, 96%) as a colorless solid, which was used in the next step without further purification.

[0220] Example 1.18 - (2R,3S,4R,5S,6R)-2-(Acetoxymethyl)-5-pivalamido-6-propyltetrahydro-2H-pyran-3,4-diyl diacetate (28):

[0221] To a solution of 27 (32 mg, 0.11 mmol) and acetic anhydride (0.08 mL, 0.83 mmol) in pyridine (0.65 mL) was added a catalytic amount of DMAP. The solution was stirred at room temperature for 16 h and diluted with ethyl acetate (7.5 mL). The organic phase was washed sequentially with 1M HCl (3x5 mL), water (1x 5 mL), saturated aqueous NaHCO3 (1x 5 mL), brine (5 mL x 1), then dried (Na2SO4) and concentrated under reduced pressure. The resulting residue was purified by chromatography on SiO2 (hexane / EtOAc, 4:1) to afford 28 (34 mg, 0.082 mmol, 74%) as a colorless amorphous solid: HRMS m / z calculated for C 20 H 34 O8N([M+1] + )416.2279, found 416.2309;1 H NMR(400MHz, CDCl3) δ6.06(d,J=8.0Hz,1H),5.05-5.01(m,1H),4.98-4.94(m,1H),4.36(dd,J=12.0,6.4Hz,1H),4.24-4.20(m,1H),4.17-4.0 1(m,2H),3.86-3.84(m,1H),2.09(s,3H),2.08(s,3H),2.07(s,3H),1. 65-1.55(m,2H),1.35-1.25(m,2H),1.18(s,9H),0.93(t,J=7.2Hz,3H).

[0222] This example further provides the following suggested syntheses: (2R,3S,4R,5S,6R)-6-allyl-2-((butyryloxy)methyl)-5-pivaloylaminotetrahydro-2H-pyran-3,4-diyl dibutyrate (31), (2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-isobutyl-5-pivaloylaminotetrahydro-2H-pyran-3,4-diyl diacetate (32), (1S,2S,4S,6S)-2-acetoxy-4-(acetoxymethyl)-6-propoxycyclohexyl pivalate (33), N-((2S,3R,4R,5S,6R)-

[00136] 4,5-dihydroxy-6-(hydroxymethyl)-2-isopropyltetrahydro-2H-pyran-3-yl)pivalamide (34), (2R,3S,4R,5S,6R)-6-allyl-2-((benzoyloxy)methyl)-4-(2-phenylacetyloxy)-5-pivalamidotetrahydro-2H-pyran-3-ylbenzoate (35), and (2R,3S,4R,5S,6R)-2-(((azetidine-1-carbonyl)oxy)methyl)-5-pivalamido-6-propyltetrahydro-2H-pyran-3,4-diylbis(azetidine-1-carboxylate) (36), as shown in Scheme VI.

[0223]

[0224] Example 1.19 - (2R,3S,4R,5S,6R)-6-allyl-2-((butyryloxy)methyl)-5-pivalamidotetrahydro-2H-pyran-3,4-diyl dibutyrate (31):

[0225] Starting with 25, a synthesis similar to Scheme V was used to give 31. Using deprotection conditions, the olefin can be retained in the debenzylation step (see, e.g., Cavedon, C.; Sletten, ET; Madani, A.; Niemeyer, O.; Seeberger, PH; Pieber, B. Org. Lett. 2021, 23(2), 514-518).

[0226] Example 1.20 - (2R, 3S, 4R, 5S, 6R)-2-(Hydroxymethyl)-6-isobutyl-5-pivalamidotetrahydro-2H-pyran-3,4-diyl diacetate (32):

[0227] Starting with 29, a synthesis analogous to Scheme V was used. Further steps using the product of step 3 can be achieved by selective deprotection at the C-6 hydroxyl using methods disclosed by, for example, Filice et al. (See, Filice, M.; Guisan, JM; Terreni, M.; Palomo, JM Nature Protocols. 2012, 7(10), 1783-1796.) to give 32.

[0228] Example 1.21 - (2R, 3S, 4R, 5S, 6R)-2-(Hydroxymethyl)-5-pivalamido-6-propyltetrahydro-2H-pyran-3,4-diyl diacetate (33):

[0229] Starting with 25, a synthesis analogous to Scheme V was used. Further steps using the product of step 3 can be achieved by selective deprotection at the C-6 hydroxyl using, for example, the method disclosed by Filice et al. (see Filice, M.; Guisan, JM; Terreni, M.; Palomo, JM Nature Protocols. 2012, 7(10), 1783-1796.) to give 33.

[0230] Example 1.22 - N-((2S,3R,4R,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-isopropyltetrahydro-2H-pyran-3-yl)pivalamide (34):

[0231] Starting with 30, a synthesis analogous to steps 1-2 of Scheme V was used to obtain 34 (see, eg, Wipf, P.; Pierce, JG; Zhuang, N. Org. Lett. 2005, 7(3), 483-485.).

[0232] Example 1.23 - (2R, 3S, 4R, 5S, 6R)-6-allyl-2-((benzoyloxy)methyl)-4-(2-phenylacetoxy)-5-pivalamidotetrahydro-2H-pyran-3-ylbenzoate (35):

[0233] Starting with 25, a synthesis similar to Scheme V is used. Deprotection conditions can be used to retain the olefin in the debenzylation step (see, e.g., Cavedon, C.; Sletten, ET; Madani, A.; Niemeyer, O.; Seeberger, PH; Pieber, B. Org. Lett. 2021, 23(2), 514-518). Further steps using the product of step 3 can be achieved by selective deprotection followed by selective protection at the C-6 hydroxyl using, for example, the methods disclosed by Filice et al. (see Filice, M.; Guisan, JM; Terreni, M.; Palomo, JM Nature Protocols. 2012, 7(10), 1783-1796.) to give 35.

[0234] Example 1.24 - (2R,3S,4R,5S,6R)-2-(((azetidine-1-carbonyl)oxy)methyl)-5-pivalamido-6-propyltetrahydro-2H-pyran-3,4-diylbis(azetidine-1-carboxylate) (36):

[0235] Starting with 25, a synthesis similar to Scheme V was used. Step 3 was accomplished using, for example, the method disclosed in WO 2017 / 023631 A1 using 1-azetidinecarbonyl chloride to give 36.

[0236] This example further provides the synthesis of (1R,2S,3S)-5-(((3,3-dimethylbutanoyl)oxy)methyl)-3-propoxycyclohexane-1,2-diyl bis(3,3-dimethylbutyrate) (42), as shown in Scheme VII.

[0237]

[0238] Example 1.25 - (3aR,7S,7aR)-7-(allyloxy)-2,2-dimethyl-3a,4,7,7a-tetrahydrobenzo[d][1,3]dioxole-5-carboxylic acid methyl ester (38):

[0239] To a solution of 37 (230 mg, 1.01 mmol) in toluene (3.6 mL) was added TlOEt (0.1 mL) at room temperature. After 2 h, the reaction mixture was concentrated under reduced pressure, and a solution of the residue in MeCN (3.6 mL) was treated with allyl iodide (0.3 mL) and stirred in the dark. After 16 h, the mixture was purified by The solids were removed by filtration, the filter pad was rinsed with CH2Cl2, the filtrate was concentrated under reduced pressure, and the residue was purified by chromatography on SiO2 (hexanes / EtOAc, 2:3) to afford 46 mg of 38 as an orange oil: 1 HNMR(CDCl3,400MHz)δ7.04(s,1H),6.01-5.91(m,1H),5.34-5.29(m,1H),5.25-5.22(dd,J=10.4,1.2Hz,1H),4.67-4.64(m,1H),4.62-4.59(m, 1H),4.30-4.25(m,1H),4.20-4.14(m,1H),3.87-3.85(m,1H),3.77(s,3 H),3.03(d,J=16.3Hz,1H),1.94-1.84(m,1H),1.32(s,3H),1.31(s,3H); 13 C NMR (CDCl3, 400 MHz) δ 166.2, 140.8, 134.4, 129.0, 118.3, 109.2, 75.7, 75.2, 73.0, 71.1, 52.1, 27.5, 26.0, 24.3; HRMS m / z calculated for C 14 H 21 O5([M+H] + )269.1407, found 269.1407 (see, e.g., Mohanrao, R.; Asokan, A.; Sureshan, KM Chem. Commun. 2014, 50(51), 6707-6710.).

[0240] Example 1.26 - ((3aR,7S,7aR)-7-(allyloxy)-2,2-dimethyl-3a,4,7,7a-tetrahydrobenzo[d][1,3]dioxol-5-yl)methanol (39):

[0241] At -78 ° C, a solution of methyl ester 38 (0.040 g, 0.149 mmol, 1 equivalent) in THF (0.2 mL) was treated with DIBAL-H (0.298 mL, 0.298 mmol, 2 equivalents, 1 mmol, 1 M in hexane), warmed to room temperature over several hours, and stirred overnight. The reaction mixture was quenched with saturated ammonium chloride (5 mL), then extracted with CH Cl (3 x 10 mL). The combined organic layers were dried (MgSO ) and concentrated in vacuo to give 39 (0.015 g, 0.062 mmol, 42%), which was used for subsequent steps without further purification.

[0242] Example 1.27 - (1R,2R,3S)-3-(allyloxy)-5-(hydroxymethyl)cyclohex-4-ene-1,2-diol (40):

[0243] A solution of 39 (0.038 g, 0.158 mmol) in TFA and water (80:20, 0.53 mL) was stirred at room temperature for 30 min and concentrated in vacuo to afford 40 (0.011 g, 0.055 mmol, 88%) which was used in the subsequent step without further purification.

[0244] Example 1.28 - (1R,2R,3S)-3-(allyloxy)-5-(((3,3-dimethylbutanoyl)oxy)methyl)cyclohex-4-ene-1,2-diyl bis(3,3-dimethylbutyrate) (41):

[0245] A solution of 40 (0.080 g, 0.400 mmol, 1 eq) in pyridine (1.33 mL) was treated with tert-butylacetyl chloride (0.566, 4.00 mmol, 10 eq) and catalytic DMAP and stirred at room temperature overnight. The reaction mixture was quenched by adding EtOAc (5 mL), extracted with 1 M HCl (2 x 5 mL) and brine (5 mL), dried (MgSO4), and concentrated under reduced pressure. The resulting residue was purified by chromatography on SiO2 (8: 1, hexane: EtOAc) to give 41 (0.087 g, 0.176 mmol, 71%) as a colorless oil: 1 H NMR(CDCl3,300MHz)δ5.82-5.79(m,1H),5.66-5.61(m,2H),5.29-5.02(m,3H),4.49(bs,2H),4.19 -4.11(m,2H),4.00-3.92(m,1H),2.38-2.31(m,2H),2.24(s,2H),2.22(s,2H),2.17(s,2H),1.03(s 9H),1.02(s,9H),1.00(s,9H).

[0246] Example 1.29 - (1R, 2S, 3S)-5-(((3,3-dimethylbutanoyl)oxy)methyl)-3-propoxycyclohexane-1,2-diyl bis(3,3-dimethylbutyrate) (42):

[0247] A solution of 41 (0.020 g, 0.04 mmol, 1 eq) in THF (0.2 mL) was treated with Pd / C (10%, 0.004 g, 0.004 mmol) and H2 gas and stirred at room temperature overnight. Filtration and removal of the solvent under reduced pressure gave 42 as a 2:1 mixture of diastereomers.

[0248] This example further provides the following suggested syntheses: (1S,2S,3S,4R,6S)-4-(acetoxymethyl)-6-propoxycyclohexane-1,2,3-triyl triacetate (43), (1S,2S,4S,6S)-2-acetoxy-4-(acetoxymethyl)-6-propoxycyclohexyl pivalate (44), and (1R,2R,4S,6S)-2-hydroxy-6-propoxycyclohexane-1,4-diyl bis(2,2-dimethylpropionate) (45), as shown in Scheme VIII.

[0249]

[0250] Example 1.30 - (1S,2S,3S,4R,6S)-4-(acetoxymethyl)-6-propoxycyclohexane-1,2,3-triyl triacetate (43):

[0251] Starting with 37, a synthesis similar to Scheme VII was used to obtain 43 (see, eg, Mohanrao, R.; Asokan, A.; Sureshan, KM Chem. Commun. 2014, 50(51), 6707-6710.).

[0252] Example 1.31 - (1R, 2R, 4S, 6S)-2-Hydroxy-6-propoxycyclohexane-1,4-diyl bis(2,2-dimethylpropionate) (44):

[0253] Starting with 37, a synthesis similar to Scheme VII was used to obtain 44 (see, eg, Mohanrao, R.; Asokan, A.; Sureshan, KM Chem. Commun. 2014, 50(51), 6707-6710.).

[0254] Example 1.32 - (1S, 2S, 4S, 6S)-2-acetoxy-4-(acetoxymethyl)-6-propoxycyclohexyl pivalate (45):

[0255] Starting with 37, a synthesis similar to Scheme VII was used to obtain 45 (see, eg, Mohanrao, R.; Asokan, A.; Sureshan, KM Chem. Commun. 2014, 50(51), 6707-6710.).

[0256] This example further provides suggested syntheses of ((1S,3S,4S,5S)-3-acetoxy-5-isobutoxy-4-pivaloylaminocyclohexyl)methyl acetate (47), ((1R,3R,4S,5S)-3-acetoxy-5-phenethoxy-4-pivaloylaminocyclohexyl)methyl acetate (48), and (1R,2S,3S,5R)-2-acetylamino-3-isopropoxy-5-((pivaloyloxy)methyl)cyclohexylazetidine-1-carboxylate (49) as shown in Scheme VIII.

[0257]

[0258] Example 1.33 - ((1S,3S,4S,5S)-3-acetoxy-5-isobutoxy-4-pivaloylaminocyclohexyl)methyl acetate (47):

[0259] Starting with 46, a synthesis analogous to Scheme VII was used to obtain 47 (see, e.g., Shie, J.-J.; Fang, J.-M.; Wang, S.-Y.; Tsai, K.-C.; Cheng, Y.-SE; Yang, A.-S.; Hsiao, S.-C.; Su, C.-Y.; Wong, C.-HJ Am. Chem. Soc. 2007, 129(39), 11892-11893.).

[0260] Example 1.34 - ((1R,3R,4S,5S)-3-acetoxy-5-phenethoxy-4-pivaloylaminocyclohexyl)methyl acetate (48):

[0261] Starting with 46, a synthesis analogous to Scheme VII was used to obtain 48 (see, e.g., Shie, J.-J.; Fang, J.-M.; Wang, S.-Y.; Tsai, K.-C.; Cheng, Y.-SE; Yang, A.-S.; Hsiao, S.-C.; Su, C.-Y.; Wong, C.-HJ Am. Chem. Soc. 2007, 129(39), 11892-11893.).

[0262] Example 1.36 - (1R, 2S, 3S, 5R)-2-Acetylamino-3-isopropoxy-5-((pivaloyloxy)methyl)cyclohexylazetidine-1-carboxylate (49):

[0263] Starting with 46, a synthesis analogous to Scheme VII was used to obtain 49 (see, e.g., Shie, J.-J.; Fang, J.-M.; Wang, S.-Y.; Tsai, K.-C.; Cheng, Y.-SE; Yang, A.-S.; Hsiao, S.-C.; Su, C.-Y.; Wong, C.-HJ Am. Chem. Soc. 2007, 129(39), 11892-11893.).

[0264] Example 2: Compounds for controlling inflammatory and immune responses associated with infection

[0265] This example provides methods of controlling an inflammatory response according to certain embodiments of the present disclosure.

[0266] Experimental methods and materials

[0267] Mouse enteroids: Primary intestinal crypt cultures (enteroids) were generated from the ileum of newborn (p7-p11) mice and maintained in Matrigel (Corning). Enteroids were digested and passaged weekly using TrypLE Express (Gibco) and used between passages 3 and 10 for all experiments. Enteroids were pretreated with C34 analogs (20 μM, overnight) and then treated with LPS (50 μg / mL) for 4 hours for further analysis. Toll-like receptor 4 (Tlr4) ( Figure 1 ) and tumor necrosis factor (Tnf) ( Figure 2 )'s relative mRNA expression levels.

[0268] Mice: C57BL / 6J mice were purchased from Jackson Laboratory. All mice were housed in a specific pathogen-free environment (ambient temperature between 20 and 25 °C and humidity between 30 and 70%) with a 12-h light / 12-h dark cycle and free access to water and standard rodent chow (Teklad global, 18% protein rodent diet, Envigo).

[0269] Endotoxemia model: Endotoxemia was induced in newborn pups (p11-p14) of either sex by intraperitoneal administration of 5 mg / kg lipopolysaccharide (LPS), which were randomly divided into a control group and a test group, and ileal samples were collected 6 h after LPS treatment. C34 and C34 analogs were administered by oral gavage 1 day before LPS injection at a dose of 20 mg / kg body weight.

[0270] Mouse NEC Model: Experimental NEC was induced in 7-day-old mice of either sex using a well-validated and reproducible model. Mice were randomized into control and test groups and gavaged with a formula containing Similac Advance infant formula (Abbott Nutrition): Esbilac (PetAg) canine milk replacer in a 2:1 ratio, supplemented with intestinal bacteria prepared from a stock solution obtained from specimens of infants with surgical NEC, 5 times daily. In addition, mice were subjected to hypoxia (5% O2–95% N2) for 10 minutes twice daily for 4 days in a hypoxia chamber (Billups-Rothenberg). During the induction of NEC, C34 analogs were administered by oral gavage at a dose of 10 mg per kg body weight per day. Age-matched breast-fed mouse pups were used as non-NEC controls. Toll-like receptor 4 (Tlr4) ( Figure 3 )、Interleukin 6 (Il6) ( Figure 4 )、Interleukin 1β(Il1b)( Figure 5 ), tumor necrosis factor (Tnf) ( Figure 6 ), and lipocalin 2 (Lcn2) ( Figure 7 )'s relative mRNA expression levels.

[0271] RNA isolation, cDNA synthesis and qPCR: Total RNA was isolated using the RNeasy mini kit (Qiagen), and complementary DNA was synthesized from 0.5 μg RNA using the QuantiTect Reverse Transcription Kit (Qiagen) according to the manufacturer's protocol. iTaq TM General mRNA quantification was performed using Bio-Rad Green Supermix (Bio-Rad), and data from qRT-PCR were collected using Bio-Rad CFX Manager 3.1 software. Toll-like receptor 4 (Tlr4) ( Figure 8 )、Interleukin 6 (Il6) ( Fig. 9 )、Interleukin 1β(Il1b)( Fig.10 ), tumor necrosis factor (Tnf) ( Fig.11 ), and lipocalin 2 (Lcn2) ( Fig.12 ) were normalized to the expression of the housekeeping gene ribosomal protein lateral handle subunit P0 (Rplp0).

[0272] Hematoxylin and eosin (H&E) staining: 5 μm paraformaldehyde-fixed paraffin-embedded tissue sections were rehydrated and stained in hematoxylin solution (Sigma-Aldrich) and in Epredia TM Richard-Allan Scientific TM Differentiation solution (Fisher Scientific) was used for differentiation. TM Signature Series TM The slides were treated with bluing reagent (FisherScientific), stained with eosin Y solution (Sigma-Aldrich), dehydrated, and mounted with Permount mounting medium (Fisher Chemical) before imaging using a Leica DMi 8 microscope. Representative images of H&E staining of non-NEC control, NEC, and NEC+C281 groups are shown ( Fig.13 ).

[0273] Statistical analysis: Transcriptional levels of inflammatory biomarkers were assessed by performing one-way ANOVA statistical analysis followed by multiple comparisons. Statistical significance is indicated by p-values, where *p<0.05, ***p<0.001, ****p<0.0001. Data are expressed as relative mRNA expression normalized to the housekeeping gene Rplp0. Each data point represents a single mouse.

[0274] result

[0275] The examples disclosed herein demonstrate the effectiveness of C34 analogs in controlling inflammation and immune responses associated with infection. In vitro studies were performed using primary intestinal crypt cultures (particularly enteroids). Enteroid pretreatment with C34 analogs was shown to control lipopolysaccharide (LPS)-induced inflammation, such as Toll-like receptor 4 (Tlr4) ( Figure 1 ) and tumor necrosis factor (Tnf) ( Figure 2 )'s relative mRNA expression levels are shown in ( Figure 2 ).

[0276] This example further evaluates the effectiveness of C34 analogs in controlling inflammation in an endotoxemia mouse model. Newborn pups were subjected to endotoxemia by intraperitoneal injection of lipopolysaccharide (LPS). The effect of C34 analogs was evaluated by pre-dosing them by oral gavage 1 day before LPS injection. Ileal samples were collected 6 hours after LPS treatment to evaluate the inflammatory response marker Toll-like receptor 4 (Tlr4) ( Figure 8 )、interleukin 6 (Il6) ( Fig. 9 )、Interleukin 1β(Il1b)( Fig.10 ), tumor necrosis factor (Tnf) ( Fig.11 ), and lipocalin 2 (Lcn2) ( Fig.12 ).

[0277] Similarly, qRT-PCR analysis was performed in a mouse model targeting the inflammatory marker Toll-like receptor 4 (Tlr4) ( Figure 3 )、interleukin 6 (Il6) ( Figure 4 )、Interleukin 1β(Il1b)( Figure 5 ), tumor necrosis factor (Tnf) ( Figure 6 ) evaluated C34 analogs after induction of NEC and evaluated lipocalin 2 (Lcn2) ( Figure 7 Notably, the C281 analog exhibited a robust response. Furthermore, treatment with the C281 analog was shown to preserve the histological structure of the intestinal lining of NEC mice.

[0278] Although the disclosed subject matter and its advantages have been described in detail, it should be understood that various changes, substitutions and modifications may be made herein without departing from the spirit and scope of the disclosed subject matter. In addition, the scope of the present application is not intended to be limited to the specific embodiments of the processes, machines, manufactures and material compositions, methods and processes described in the specification. Those of ordinary skill in the art will easily understand from the disclosed subject matter of the disclosed subject matter that the processes, machines, manufactures, material compositions, methods or steps currently existing or to be developed in the future with the corresponding embodiments of the disclosed subject matter or achieving substantially the same results can be utilized according to the disclosed subject matter herein. Therefore, the appended claims are intended to include such processes, machines, manufactures, material compositions, methods or steps within their scope.

[0279] Various patents, patent applications, publications, product descriptions, protocols, and sequence accession numbers are cited throughout this application, the disclosures of which are incorporated herein by reference in their entirety for all purposes.

Claims

1. A compound of formula I or a stereoisomer thereof: wherein X is O, Z is O or NH, Y is CH2 or CH(R1), R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2, or wherein X is CH2, Z is O or NH, and Y is O, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, N(R1)2.

2. A pharmaceutical composition comprising the compound according to claim 1.

3. The pharmaceutical composition of claim 2, wherein the pharmaceutical composition is selected from the group consisting of coated particles, micelles, liposomes, tablets, capsules, sachets, suppositories, liquid pharmaceutical compositions, and combinations thereof.

4. The pharmaceutical composition of claim 2, wherein the pharmaceutical composition further comprises: an antibiotic agent, a steroidal compound, or a non-steroidal anti-inflammatory agent, or a combination thereof.

5. The compound of claim 1, wherein the compound is selected from: and combinations thereof.

6. A compound of formula II or a stereoisomer thereof: wherein Z is O or NH, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

7. A pharmaceutical composition comprising the compound according to claim 6.

8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition is selected from the group consisting of coated particles, micelles, liposomes, tablets, capsules, sachets, suppositories, liquid pharmaceutical compositions, and combinations thereof.

9. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition further comprises an antibiotic agent, a steroidal compound, or a nonsteroidal anti-inflammatory agent, or a combination thereof.

10. The compound of claim 6, wherein the compound is selected from the group consisting of: and combinations thereof.

11. A method for treating an infectious or inflammatory disorder, the method comprising administering to a subject in need of such treatment an effective amount of a Toll-like receptor 4 inhibitor compound of Formula I or a stereoisomer thereof: wherein X is O, Z is O or NH, Y is CH2 or CH(R1), R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2, or wherein X is CH2, Z is O or NH, and Y is O, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

12. A method for treating an infectious or inflammatory disorder, the method comprising administering to a subject in need of such treatment an effective amount of a Toll-like receptor 4 inhibitor compound of Formula II or a stereoisomer thereof: wherein Z is O or NH, R1 is alkyl, branched alkyl, cycloalkyl, aryl, or substituted aryl, and R2 is H or COR3, wherein R3 is selected from R1, NHR1, or N(R1)2.

13. The method of claim 11 or 12, wherein the subject suffers from one or more of the following: osteoarthritis, pancreatitis, metabolic syndrome, trauma-induced systemic inflammation, acute respiratory distress syndrome, COVID-19-induced systemic inflammation.

14. The method of claim 11 or 12, wherein the subject has necrotizing enterocolitis.

15. The method of claim 1, wherein the Toll-like receptor 4 inhibitor is selected from: and combinations thereof.

16. The method of claim 12, wherein the Toll-like receptor 4 inhibitor is selected from: and combinations thereof.

17. A method for treating traumatic injury in a subject, the method comprising administering to a subject in need of such treatment an effective amount of a Toll-like receptor 4 inhibitor compound according to claim 11 or 12, which reduces Toll-like receptor 4-induced post-traumatic injury.

18. The method of claim 17, wherein the traumatic injury is to an organ selected from the group consisting of: heart, liver, lung, kidney, intestine, brain, eye, and pancreas.

19. The method of claim 11 or 12, wherein the compound is administered to treat organ rejection following transplantation.

20. The method of claim 11 or 12, comprising administering a compound of Formula I or Formula II to prevent necrotizing enterocolitis in premature infants.

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