Broad spectrum aminoglycosides and their uses

Abstract

Provided herein are aminoglycoside compounds useful as therapeutic and / or prophylactic agents, such as compounds of formula (I) and formula (III), and pharma- ceutically acceptable salts thereof. The compounds may be useful for treating bacterial infections, such as gram-negative bacterial infections, in subjects. Methods for preparing such compounds are also provided herein. TIFF2025509208000208.tif88168

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 316,731, filed March 4, 2022, which is incorporated herein by reference in its entirety. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under Contract No. 75N93020C00018 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0002] FIELD OF THE DISCLOSURE This disclosure relates generally to aminoglycoside compounds with antibacterial activity and their use in treating medical conditions associated with pathogenic microorganisms. [Background technology]

[0003] The rapid spread of antibiotic resistance has prompted a continuous search for new agents with antibacterial activity. Aminoglycosides (AGs) are highly potent, broad-spectrum antibiotics with many desirable properties for treating life-threatening infections. AGs are used to treat a variety of diseases, including, but not limited to, urinary tract infections, bloodstream infections, pneumonia, and neonatal sepsis. Examples of AGs include tobramycin, gentamicin, and amikacin. AG antibiotics exert their antibacterial effects by binding to specific target sites in bacterial ribosomes. For the structurally related antibiotics neamine, ribostamycin, neomycin B, and paromomycin, the binding site is localized to the A site of the prokaryotic 16S ribosomal decoding region RNA (see Moazed, D.; Noller, H.F., Nature, 1987, 327, 389). Binding of AG to this RNA target interferes with the fidelity of mRNA translation, causing miscoding and truncation, ultimately leading to bacterial cell death (see Alper et al., J. Am. Chem. Soc., 1998, 120, 1965).

[0004] Over time, resistance to antimicrobials has emerged in hospitals, undermining our antimicrobial armamentarium. Antimicrobial resistance (AG-R) can be mechanistically classified into two main types: (1) AG-modifying enzymes (AMEs), which modify drugs, rendering them inactive, and (2) enzymes called ribosomal methyltransferases (RMTs), which modify the drug target binding pocket. Currently, the most common form of AG-R among clinical isolates is conferred by diverse and widely distributed AMEs. While less common than AMEs in some species, RMTs pose a serious threat because they modify the 16S rRNA AG-binding site, preventing AG target binding and conferring high-level resistance to all clinically available parenteral AGs. Of further concern, AME and RMT genes are typically located on plasmids or transposons together with genes encoding resistance to other classes of antimicrobial agents, thereby giving rise to multidrug-resistant isolates.

[0005] Furthermore, AGs can be toxic to the kidneys (nephrotoxicity) and cochlea (ototoxicity). These are cumulative processes, and increasing toxicity is likely to be observed with treatment duration. The potential for AG nephrotoxicity limits the dosage and length of treatment and increases the difficulty of achieving the systemic exposure required for efficacy against some infections. Quite frequently, but not always, the structure-activity relationships that result in increased antibacterial efficacy are associated with those that cause toxicity to human kidney cells. The origin of this toxicity is hypothesized to result from a combination of different factors and mechanisms, such as interactions with phospholipids, inhibition of phospholipases, and the formation of free radicals. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Moazed, D.; Noller, HF, Nature, 1987, 327, 389 [Non-patent document 2] Alper et al., J. Am. Chem. Soc., 1998, 120, 1965 Summary of the Invention [Problem to be solved by the invention]

[0007] For the reasons stated above, although progress has been made in this field, new chemical entities are needed that have antibacterial activity against highly resistant bacteria and an acceptable safety profile. [Means for solving the problem]

[0008] (Summary of the Invention) The present disclosure relates to aminoglycoside compounds having antibacterial activity, including their stereoisomers, tautomers, solvates and pharmaceutically acceptable salts, for example, for use in the treatment of medical conditions associated with pathogenic microorganisms, also referred to herein as "bacterial infections."

[0009] In one embodiment, formula (I)

[0010] [ka] [In the formula, R a is hydrogen (H) or methyl, R b is C 1~6 alkyl, or of formula (II)

[0011] [ka] The structure is Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3Alkyl or -C(=NH)NR 4a R 4b and R 4a and R 4b is H and C 1~3 alkyl, or R 1 and R 4 taken together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N, or a pharmaceutically acceptable salt or solvate thereof.

[0012] In some embodiments, R a is H.

[0013] In some embodiments, R a is methyl (CH3).

[0014] In some embodiments, R b teeth,

[0015] [ka] is selected from the group consisting of:

[0016] In some embodiments, R b teeth,

[0017] [ka] is selected from the group consisting of:

[0018] In some embodiments, the compound of formula (I) is

[0019] [ka] has a structure selected from the group consisting of:

[0020] In some embodiments, R bis ethyl (C2H5).

[0021] In some embodiments, the compound of formula (I) has the structure

[0022] [ka] It has.

[0023] In another embodiment, the compound of formula (III)

[0024] [ka] [In the formula, R a is H or methyl, R b is H, C 1~6 alkyl, or of formula (II)

[0025] [ka] The structure is Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR 4a R 4b and R 4a and R 4b is hydrogen and C 1~3 alkyl, or R 1 and R 4together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N.

[0026] In some embodiments, R a is H.

[0027] In some embodiments, R a is methyl.

[0028] In some embodiments, the compound of formula (III) is

[0029] [ka] has a structure selected from the group consisting of:

[0030] In some embodiments, R b is ethyl.

[0031] In some embodiments, the compound of formula (III) is

[0032] [ka] has a structure selected from the group consisting of:

[0033] In some embodiments, R b teeth,

[0034] [ka] is selected from the group consisting of:

[0035] In some embodiments, R b teeth,

[0036] [ka] is selected from the group consisting of:

[0037] In some embodiments, the compound of formula (III) is

[0038] [ka] has a structure selected from the group consisting of:

[0039] In another aspect, there is provided a pharmaceutical composition comprising a compound of Formula (I), Formula (III), or a pharmaceutically acceptable salt or solvate of either thereof, and at least one pharmaceutically acceptable excipient.

[0040] In a further aspect, there is provided a method for treating a bacterial infection in a subject in need thereof, the method comprising administering to the subject a compound of any of Formulas (I) or (III), or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising said compound. In some embodiments, the method comprises administering to the subject a compound of Formula (I) or (III), or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising said compound.

[0041] In some embodiments, the bacterial infection is caused by a gram-positive bacterium, a gram-negative bacterium, an aerobic bacterium, or a facultative anaerobic bacterium. In some embodiments, the bacterial infection is a gram-negative bacterial infection. In some embodiments, the bacterial infection is a gram-positive bacterial infection.

[0042] In some embodiments, the bacterial infection is caused by a species of Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, or the like. Infections caused by species of the genera Citrobacter, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Burkholderia, or Enterococcus.

[0043] In some embodiments, the bacterial infection is selected from the group consisting of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, Proteus mirabilis, Bacillus anthracis, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus vulgaris, and the like. Infections caused by Burkholderia vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei, or Yersinia pestis.

[0044] In some embodiments, the bacterial infection is a gram-negative bacterial infection, ie, an infection caused by Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxidans.

[0045] In some embodiments, the bacterial infection is caused by a multidrug-resistant (MDR) bacteria.

[0046] In some embodiments, the bacterial infection is an infection caused by extended-spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae.

[0047] In some embodiments, the bacterial infection is caused by Staphylococcus aureus. In some embodiments, the Staphylococcus aureus is methicillin-resistant Staphylococcus aureus (MRSA).

[0048] The present disclosure includes, but is not limited to, the following embodiments.

[0049] Embodiment 1. Formula (I)

[0050] [ka] [In the formula, R a is hydrogen (H) or methyl, R b is C 1~6 alkyl, or of formula (II)

[0051] [ka] The structure is Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR 4a R 4b and R4a and R 4b is H and C 1~3 alkyl, or R 1 and R 4 or a pharmaceutically acceptable salt or solvate thereof.

[0052] Embodiment 2: R a is H.

[0053] Embodiment 3: R a is methyl.

[0054] Embodiment 4: R b but,

[0055] [ka] The compound of any one of embodiments 1-3, selected from the group consisting of:

[0056] Embodiment 5: R b but,

[0057] [ka] The compound of any one of embodiments 1-3, selected from the group consisting of:

[0058] Embodiment 6: The compound of Formula (I) is

[0059] [ka] The compound of embodiment 1, having a structure selected from the group consisting of:

[0060] Embodiment 7: R bThe compound of any one of embodiments 1-3, wherein is ethyl.

[0061] Embodiment 8: The compound of Formula (I) has the structure

[0062] [ka] 8. The compound of embodiment 7, having the formula:

[0063] Embodiment 9: Formula (III)

[0064] [ka] [In the formula, R a is H or methyl, R b is H, C 1~6 alkyl, or of formula (II)

[0065] [ka] The structure is Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR 4a R 4b and R 4a and R 4b is hydrogen and C 1~3 alkyl, or R 1 and R 4together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N.

[0066] Embodiment 10:R a is H.

[0067] Embodiment 11:R a is methyl.

[0068] Embodiment 12: The compound of formula (III) is

[0069] [ka] 12. The compound of any one of embodiments 9-11, having a structure selected from the group consisting of:

[0070] Embodiment 13:R b The compound of any one of embodiments 9-12, wherein is ethyl.

[0071] Embodiment 14: The compound of formula (III) is

[0072] [ka] The compound of embodiment 13, having a structure selected from the group consisting of:

[0073] Embodiment 15:R b but,

[0074] [ka] 13. The compound of any one of embodiments 9-12, selected from the group consisting of:

[0075] Embodiment 16:R b but,

[0076] [ka] 13. The compound of any one of embodiments 9-12, selected from the group consisting of:

[0077] Embodiment 17: The compound of formula (III) is

[0078] [ka] 17. The compound of embodiment 16, having a structure selected from the group consisting of:

[0079] Embodiment 18: A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 17 or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof and at least one pharmaceutically acceptable excipient.

[0080] Embodiment 19: A method for treating a bacterial infection in a subject in need thereof, comprising administering to the subject a compound of any one of embodiments 1 to 17 or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or a pharmaceutical composition of embodiment 18.

[0081] Embodiment 20: The method of embodiment 19, wherein the bacterial infection is caused by a gram-positive bacterium, a gram-negative bacterium, an aerobic bacterium, or a facultative anaerobic bacterium.

[0082] Embodiment 21: The method of embodiment 19, wherein the bacterial infection is an infection with a species of the genus Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Burkholderia, or Enterococcus.

[0083] Embodiment 22: The method of embodiment 20 or 21, wherein the bacterial infection is an infection caused by Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, Proteus mirabilis, Bacillus anthracis, P. aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei, or Yersinia pestis.

[0084] Embodiment 23: The method of any one of embodiments 19 to 22, wherein the bacterial infection is a gram-negative bacterial infection.

[0085] Embodiment 24: The method of embodiment 23, wherein the Gram-negative bacterial infection is an infection caused by Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxidans.

[0086] Embodiment 25: The method of any one of embodiments 19 to 24, wherein the bacterial infection is caused by a multidrug-resistant (MDR) bacterium.

[0087] Embodiment 26: The method of any one of embodiments 19 to 22, wherein the bacterial infection is an infection caused by an extended-spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae group.

[0088] Embodiment 27: The method of any one of embodiments 19 to 25, wherein the bacterial infection is caused by Staphylococcus aureus.

[0089] Embodiment 28: The method of embodiment 27, wherein the Staphylococcus aureus is methicillin-resistant Staphylococcus aureus (MRSA).

[0090] These and other features, aspects, and advantages of the present disclosure will become apparent upon reading the following detailed description. The present disclosure includes any combination of two, three, four, or more features or elements specified in the present disclosure, regardless of whether such features or elements are explicitly combined or otherwise listed in a specific embodiment implementation described herein. The present disclosure is intended to be read holistically, such that any separable features or elements of the present disclosure should be considered combinable in both aspects and embodiment implementations unless the context of the disclosure clearly dictates otherwise.

[0091] Therefore, it will be appreciated that this Summary of the Invention has been provided solely for the purpose of summarizing some exemplary implementations in order to provide a basic understanding of some aspects of the present disclosure. Accordingly, it will be appreciated that the exemplary implementations described above are merely examples and should not be construed in any way to narrow the scope or spirit of the present disclosure. Other exemplary implementations, aspects, and advantages will become apparent from the following detailed description.

[0092] Having thus described aspects of the present disclosure in the foregoing general terms, reference is now made to the accompanying drawings, which are not necessarily drawn to scale, and which are illustrative only and should not be construed as limiting the disclosure. [Brief explanation of the drawings]

[0093] [Figure 1] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the structure of ring A1 according to one embodiment. [Figure 2] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the structure of ring A2 according to one embodiment. [Figure 3] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the structure of ring A3 according to one embodiment. [Figure 4] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the structure of Ring B according to one embodiment. [Figure 5] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the structure of ring AB1 according to one embodiment. [Figure 6] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the structure of ring AB2 according to one embodiment. [Figure 7] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the structure of ring AB3 according to one embodiment. [Figure 8] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the ring structure ABCD1 according to one embodiment. [Figure 9] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the ring structure ABCD2 according to one embodiment. [Figure 10] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure having the ring structure ABCD3 according to one embodiment. [Figure 11] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure according to Formula (I) having the structure of ring ABCD1 according to one embodiment. [Figure 12] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure according to formula (III) having the ring structure ABCD2 according to one embodiment. [Figure 13] FIG. 1 shows a non-limiting scheme for preparing compounds of the present disclosure according to formula (III) having the ring ABCD3 structure according to one embodiment. [Figure 14] FIG. 1 shows a non-limiting scheme for preparing a protected precursor of (2R,3R)-4-amino-3-fluoro-2-hydroxybutanoic acid according to one embodiment. [Figure 15] DETAILED DESCRIPTION OF THE INVENTION

[0094] The present disclosure will now be described in more detail with respect to example embodiments thereof. Before describing some example embodiments of the present technology, it should be understood that the technology is not limited to the details of construction or process steps set forth in the following description. The present technology is capable of other embodiments and of being practiced or carried out in various ways.

[0095] The following description sets forth numerous example arrangements, methods, parameters, etc., to provide a thorough understanding of various embodiments of the present disclosure. However, it should be understood that such description is not intended as a limitation on the scope of the present disclosure, but is instead provided as a description of example embodiments.

[0096] The present disclosure is generally directed to aminoglycoside compounds, including their stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts, and to pharmaceutical compositions comprising said compounds or their stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts. The present disclosure is particularly directed to such compounds, their stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts, that have antibacterial activity. Thus, the disclosed compounds and pharmaceutical compositions may be useful for treating bacterial infections and related medical conditions. The disclosed compounds and pharmaceutical compositions may also be useful for treating difficult-to-treat resistance mechanisms and multidrug-resistant (MDR) bacterial infections.

[0097] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications mentioned herein are incorporated by reference.

[0098] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The following definitions are provided for terms used in this disclosure. This application uses the following terms as defined below, unless the context of the sentence in which the term appears requires a different meaning.

[0099] As used in this disclosure, the articles "a" and "an" may refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" may mean one element or more than one element.

[0100] As used in this disclosure, the term "and / or" can mean either "and" or "or" unless otherwise indicated.

[0101] The term "about" used throughout this specification is used to describe and clarify slight variations. For example, the term "about" can refer to ±10% or less, or ±5% or less, such as ±2% or less, ±1% or less, ±0.5% or less, ±0.2% or less, ±0.1% or less, or ±0.05% or less. All numerical values ​​in this specification are modified by the term "about," whether or not explicitly indicated. Values ​​modified by the term "about" of course include the specific value. For example, "about 5.0" must include 5.0.

[0102] Unless the context requires otherwise, as used herein throughout this specification and the claims, the terms "including," "containing," and "comprising" are used in an open and non-limiting sense.

[0103] "Alkyl" refers to an alkyl group consisting solely of carbon and hydrogen atoms, containing no unsaturation, and preferably having 1 to 15 carbon atoms (i.e., C1 to C 15In certain embodiments, alkyl refers to a straight or branched hydrocarbon chain radical having 1 to 13 carbon atoms (i.e., C1-C 13 In some embodiments, an alkyl contains 1 to 8 carbon atoms (i.e., a C1-C8 alkyl). In other embodiments, an alkyl contains 1 to 5 carbon atoms (i.e., a C1-C5 alkyl). In other embodiments, an alkyl contains 1 to 4 carbon atoms (i.e., a C1-C4 alkyl). In other embodiments, an alkyl contains 1 to 3 carbon atoms (i.e., a C1-C3 alkyl). In other embodiments, an alkyl contains 1 to 2 carbon atoms (i.e., a C1-C2 alkyl). In some embodiments, an alkyl contains 2 carbon atoms (i.e., a C2 alkyl or ethyl). In other embodiments, an alkyl contains 1 carbon atom (i.e., a C1 alkyl). In other embodiments, an alkyl contains 5 to 15 carbon atoms (i.e., a C5-C6 alkyl). 15 In other embodiments, an alkyl contains 5 to 8 carbon atoms (i.e., a C5-C8 alkyl). In other embodiments, an alkyl contains 2 to 5 carbon atoms (i.e., a C2-C5 alkyl). In other embodiments, an alkyl contains 3 to 5 carbon atoms (i.e., a C3-C5 alkyl). In some embodiments, an alkyl group is chosen from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1,1-dimethylethyl (tert-butyl), and 1-pentyl (n-pentyl). An alkyl is attached to the remainder of the molecule by a single bond.

[0104] "C x~y The term " when used with a chemical moiety, such as alkyl, alkenyl, or alkynyl, is meant to include groups containing x to y carbons in the chain. For example, "C 1~6 The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups, containing 1 to 6 carbons. x~yThe term alkylene- refers to a substituted or unsubstituted alkylene chain having x to y carbons in the alkylene chain. For example, -C 1~6 Alkylene- may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any one of which is optionally substituted.

[0105] "Alkoxy" refers to a radical attached through an oxygen atom of the formula -o-alkyl, where alkyl is an alkyl chain as defined above.

[0106] "Alkenyl" refers to an alkyl group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and preferably having 2 to 12 carbon atoms (i.e., C2 to C6 12 "Alkenyl" refers to a straight or branched hydrocarbon chain radical group. In certain embodiments, alkenyl contains 2 to 8 carbon atoms (i.e., C2-C8 alkenyl). In certain embodiments, alkenyl contains 2 to 6 carbon atoms (i.e., C2-C6 alkenyl). In other embodiments, alkenyl contains 2 to 4 carbon atoms (i.e., C2-C4 alkenyl). Alkenyls are attached to the rest of the molecule by a single bond, and include, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like.

[0107] "Alkynyl" refers to an alkyl group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, and preferably having 2 to 12 carbon atoms (i.e., C2-C6 12 Alkynyl refers to a straight or branched hydrocarbon chain radical group. In certain embodiments, an alkynyl contains 2 to 8 carbon atoms (i.e., a C2-C8 alkynyl). In other embodiments, an alkynyl contains 2 to 6 carbon atoms (i.e., a C2-C6 alkynyl). In other embodiments, an alkynyl contains 2 to 4 carbon atoms (i.e., a C2-C4 alkynyl). An alkynyl is attached to the rest of the molecule by a single bond, and examples include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

[0108] "C x~y alkenyl" and "C x~y The term "alkynyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively. x~y The term alkenylene- refers to a substituted or unsubstituted alkenylene chain having x to y carbons in the chain. For example, -C 2~6 Alkenylene- can be selected from ethenylene, propenylene, butenylene, pentenylene, and hexenylene, any one of which is optionally substituted. The alkenylene chain can have one double bond or more than one double bond within the alkenylene chain. -C x~y The term alkynylene- refers to a substituted or unsubstituted alkynylene chain having x to y carbons in the chain. For example, -C 2~6 Alkenylene- can be selected from ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any one of which is optionally substituted. The alkynylene chain can have one triple bond or more than one triple bond within the alkynylene chain.

[0109] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain, consisting solely of carbon and hydrogen, containing no unsaturation, preferably having 1 to 12 carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, etc., that connects the rest of the molecule to the radical group. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through any two carbons within the chain. In some embodiments, alkylene contains 1 to 10 carbon atoms (i.e., C1-C8 alkylene). In some embodiments, alkylene contains 1 to 8 carbon atoms (i.e., C1-C8 alkylene). In other embodiments, alkylene contains 1 to 5 carbon atoms (i.e., C1-C5 alkylene). In other embodiments, alkylene contains 1 to 4 carbon atoms (i.e., C1-C4 alkylene). In other embodiments, alkylene contains 1 to 3 carbon atoms (i.e., C1-C3 alkylene). In other embodiments, alkylene contains 1 to 2 carbon atoms (i.e., C1-C2 alkylene). In other embodiments, alkylene contains 1 carbon atom (i.e., C1 alkylene). In other embodiments, alkylene contains 5 to 8 carbon atoms (i.e., C5-C8 alkylene). In other embodiments, alkylene contains 2 to 5 carbon atoms (i.e., C2-C5 alkylene). In other embodiments, alkylene contains 3 to 5 carbon atoms (i.e., C3-C5 alkylene).

[0110] "Alkenylene" or "alkenylene chain" refers to a straight or branched divalent hydrocarbon chain, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond, preferably having 2 to 12 carbon atoms, that connects the rest of the molecule to a radical group. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through any two carbons within the chain. In some embodiments, alkenylene contains 2 to 10 carbon atoms (i.e., C2 to C6). 10alkenylene). In certain embodiments, alkenylene contains 2 to 8 carbon atoms (i.e., C2-C8 alkenylene). In other embodiments, alkenylene contains 2 to 5 carbon atoms (i.e., C2-C5 alkenylene). In other embodiments, alkenylene contains 2 to 4 carbon atoms (i.e., C2-C4 alkenylene). In other embodiments, alkenylene contains 2 to 3 carbon atoms (i.e., C2-C3 alkenylene). In other embodiments, alkenylene contains 2 carbon atoms (i.e., C2 alkenylene). In other embodiments, alkenylene contains 5 to 8 carbon atoms (i.e., C5-C8 alkenylene). In other embodiments, alkenylene contains 3 to 5 carbon atoms (i.e., C3-C5 alkenylene).

[0111] "Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, preferably having 2 to 12 carbon atoms, that connects the rest of the molecule to a radical group. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the radical group can be through any two carbons within the chain. In some embodiments, alkynylene contains 2 to 10 carbon atoms (i.e., C2 to C6). 10 alkynylene). In certain embodiments, alkynylene contains 2 to 8 carbon atoms (i.e., C2-C8 alkynylene). In other embodiments, alkynylene contains 2 to 5 carbon atoms (i.e., C2-C5 alkynylene). In other embodiments, alkynylene contains 2 to 4 carbon atoms (i.e., C2-C4 alkynylene). In other embodiments, alkynylene contains 2 to 3 carbon atoms (i.e., C2-C3 alkynylene). In other embodiments, alkynylene contains 2 carbon atoms (i.e., C2 alkynylene). In other embodiments, alkynylene contains 5 to 8 carbon atoms (i.e., C5-C8 alkynylene). In other embodiments, alkynylene contains 3 to 5 carbon atoms (i.e., C3-C5 alkynylene).

[0112] "Aryl" refers to a radical derived from an aromatic monocyclic or aromatic polycyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. An aromatic monocyclic or aromatic polycyclic hydrocarbon ring system contains only hydrogen and carbon and 5 to 18 carbon atoms, and at least one of the rings in the ring system is aromatic, i.e., contains a cyclic delocalized (4n+2) π-electron system according to Huckel theory. Ring systems from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin, and naphthalene.

[0113] "Aralkyl" refers to a radical of the formula -R-aryl where R is an alkylene chain as defined above, e.g., methylene, ethylene, etc.

[0114] "Aralkenyl" refers to a radical of the formula -R-aryl where R is an alkenylene chain as defined above. "Aralkynyl" refers to a radical of the formula -R-aryl where R is an alkynylene chain as defined above.

[0115] "Carbocycle" refers to a saturated, unsaturated, or aromatic ring in which each atom of the ring is carbon. Carbocycles can include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. In some embodiments, a carbocycle is aryl. In some embodiments, a carbocycle is cycloalkyl. In some embodiments, a carbocycle is cycloalkenyl. In exemplary embodiments, an aromatic ring, such as phenyl, can be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings, if permitted by valence, is included in the definition of carbocycle. Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl.

[0116] "Cycloalkyl" refers to a stable, fully saturated, monocyclic or polycyclic hydrocarbon radical, consisting solely of carbon and hydrogen atoms, preferably having 3 to 12 carbon atoms, including fused or bridged ring systems. In some embodiments, a cycloalkyl contains 3 to 10 carbon atoms. In other embodiments, a cycloalkyl contains 5 to 7 carbon atoms. A cycloalkyl can be attached to the remainder of the molecule by a single bond. Examples of monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl.

[0117] "Cycloalkenyl" refers to a stable unsaturated non-aromatic monocyclic or polycyclic hydrocarbon radical, consisting solely of carbon and hydrogen atoms, preferably having from 3 to 12 carbon atoms, including fused or bridged ring systems, and containing at least one double bond. In some embodiments, a cycloalkenyl contains from 3 to 10 carbon atoms. In other embodiments, a cycloalkenyl contains from 5 to 7 carbon atoms. A cycloalkenyl can be attached to the remainder of the molecule by a single bond. Examples of monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0118] "Cycloalkylalkyl" refers to a radical of the formula -R-cycloalkyl where R is an alkylene chain as defined above.

[0119] "Cycloalkylalkoxy" refers to a group of the formula -OR c -cycloalkyl (wherein R c refers to a radical attached through an oxygen atom of the alkylene chain shown above.

[0120] "Halo" or "halogen" refers to a halogen substituent, such as bromo, chloro, fluoro, and iodo substituents.

[0121] As used herein, the term "haloalkyl" or "haloalkane" refers to an alkyl group, as defined above, that is substituted with one or more halogen groups, e.g., trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. In some embodiments, the alkyl portion of the fluoroalkyl radical is optionally further substituted. Examples of halogen-substituted alkanes ("haloalkanes") include halomethanes (e.g., chloromethane, bromomethane, fluoromethane, iodomethane), di- and trihalomethanes (e.g., trichloromethane, tribromomethane, trifluoromethane, triiodomethane), 1-haloethane, 2-haloethane, 1,2-dihaloethane, 1-halopropane, 2-halopropane, 3-halopropane, 1,2-dihalopropane, 1,3-dihalopropane, 2,3-dihalopropane, 1,2,3-trihalopropane, and any other suitable combination of alkane (or substituted alkane) and halogen (e.g., Cl, Br, F, I, etc.). When an alkyl group is substituted with more than one halogen group, each halogen can be independently selected, such as in 1-chloro, 2-fluoroethane.

[0122] "Fluoroalkyl" refers to an alkyl group, as defined above, that is substituted with one or more fluoro groups, for example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like.

[0123] "Heterocycle" refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring in a bicyclic heterocycle can be selected from saturated, unsaturated, and aromatic rings. In some embodiments, a heterocycle is heteroaryl. In some embodiments, a heterocycle is heterocycloalkyl. "Heterocyclene" refers to a divalent heterocycle that links the remainder of the molecule to a radical group.

[0124] "Heterocycloalkyl" refers to a stable 3- to 12-membered non-aromatic ring radical containing 2 to 12 carbon atoms and at least one heteroatom, each heteroatom may be selected from N, O, Si, P, B, and S atoms. Heterocycloalkyls may be selected from monocyclic or bicyclic and fused or bridged ring systems. The heteroatoms in the heterocycloalkyl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heterocycloalkyl radical is partially or fully saturated. The heterocycloalkyl is attached to the remainder of the molecule through any atom of the heterocycloalkyl, e.g., any carbon or nitrogen atom of the heterocycloalkyl, if valence allows. Examples of heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.

[0125] "Heterocycloalkylalkyl" refers to a radical of the formula -R-heterocycloalkyl where R is an alkylene chain as defined above. If the heterocycloalkyl is a nitrogen-containing heterocycloalkyl, the heterocycloalkyl is optionally attached to the alkylene chain at the nitrogen atom.

[0126] "Heteroaryl" or "aromatic heterocycle" refers to a radical derived from a 3- to 12-membered aromatic ring radical containing 1 to 11 carbon atoms and at least one heteroatom, each heteroatom being selected from N, O, and S. As used herein, heteroaryl rings may be selected from monocyclic or bicyclic ring systems and fused or bridged ring systems, where at least one of the rings in the ring system is aromatic, i.e., contains a cyclic delocalized (4n+2) π-electron system according to Huckel theory. The heteroatoms in the heteroaryl radical may be optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heteroaryl may be attached to the remainder of the molecule through any atom of the heteroaryl, e.g., a carbon or nitrogen atom of the heteroaryl, if valence allows. Examples of heteroaryl include azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, and benzotriazolyl. , benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10-Hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepam quinazolinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3 ,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2, Examples of heteroaryls include, but are not limited to, thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e., thienyl). "X-membered heteroaryl" refers to the number of endocyclic atoms, i.e., the number of Xs in the ring. For example, a 5-membered heteroaryl ring or a 5-membered aromatic heterocycle has 5 ring atoms, such as triazole, oxazole, thiophene, etc.

[0127] "Heteroarylalkyl" refers to a radical of the formula -R-heteroaryl where R is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkylene chain at the nitrogen atom.

[0128] "Isomers" are different compounds with the same molecular formula. "Stereoisomers" are isomers that differ only in the way the atoms are arranged in space. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term "(±)" is used to designate a racemic mixture when appropriate. "Diastereoisomers" or "diastereomers" are stereoisomers that have at least two asymmetric atoms but are not 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 at each chiral carbon may be specified as either R or S. Resolved compounds of unknown absolute configuration may be designated (+) or (-) depending on the direction (dextrorotatory or levorotatory) they rotate plane-polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers and may therefore give rise to enantiomers, diastereoisomers, and other stereoisomers, which asymmetric centers may be defined in terms of absolute stereochemistry as (R)- or (S)-. The present chemical compounds, pharmaceutical compositions, and methods are meant to include all such possible stereoisomers, including racemic mixtures, optically pure forms, mixtures of diastereoisomers, and intermediate mixtures. Optically active (R)- and (S)-isomers may be prepared using chiral synthons or chiral reagents or resolved using conventional techniques. The optical activity of a compound may be analyzed by any suitable method, including, but not limited to, chiral chromatography and polarimetry, and the degree of predominance of one stereoisomer over another may be determined.

[0129] In some embodiments, the compounds of the present disclosure may contain asymmetric or chiral centers and thus may exist in different stereoisomers. The term "stereoisomer" may refer to a set of compounds that have the same number and type of atoms and share the same bond connectivity between those atoms, but differ in terms of three-dimensional structure. The term "stereoisomer" may refer to any member of this set of compounds. For example, a stereoisomer may be an enantiomer or a diastereoisomer. All stereoisomers of the compounds of the present disclosure, as well as mixtures thereof, including racemic mixtures, are intended to form part of the present disclosure.

[0130] Where stereochemistry is not specified, a particular molecule described herein includes isomers, e.g., enantiomers and diastereomers, mixtures of enantiomers including racemates, mixtures of diastereomers, and other mixtures thereof, to the extent that one skilled in the art can make them using routine experimentation. In certain embodiments, single enantiomers or diastereomers, i.e., optically active forms, can be obtained by asymmetric synthesis or by resolution of a racemate or mixture of diastereomers. Resolution of a racemate or mixture of diastereomers, if possible, can be achieved by conventional methods such as, for example, crystallization in the presence of a resolving agent or chromatography, e.g., using a chiral high-pressure liquid chromatography (HPLC) column. Furthermore, mixtures of two enantiomers enriched in one of the two can be purified to obtain further optically enriched forms of the major enantiomer by recrystallization and / or trituration.

[0131] In certain embodiments, the chiral centers of the present disclosure can have the S or R configuration as defined by the IUPAC 1974 Recommendations.

[0132] As used herein, the term "amino" refers to --NH.sub.2.

[0133] As used herein, the term "hydroxy" or "hydroxyl" refers to --OH.

[0134] As used herein, the term "oxo" refers to the group "=O," which may also be abbreviated herein as C(O) or C=O.

[0135] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbon or substitutable heteroatoms, e.g., the NH of the structure. It is understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, i.e., a compound that does not undergo spontaneous transformation, e.g., by rearrangement, cyclization, elimination, and the like. In certain embodiments, substituted refers to a moiety having substituents replacing two hydrogen atoms on the same carbon atom, e.g., replacing two hydrogen atoms on a single carbon with oxo, imino, or thioxo groups. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms.

[0136] In some embodiments, a substituent may be any substituent described herein, for example, halogen, hydroxy, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO), imino (=NH), oximo (=N-OH), hydrazino (=N-NH), -R b -OR a , -R b -OC(O)-R a , -R b -OC(O)-OR a , -R b -OC(O)-N(R a )2, -R b -N(R a)2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (wherein t is 1 or 2), -R b -S(O) t R a (wherein t is 1 or 2), -R b -S(O) t OR a (wherein t is 1 or 2) and -R b -S(O) t N(R a )2 (wherein t is 1 or 2), as well as alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl, any of which may be alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO2), imino (=NH), oximo (=N-OH), hydrazine (=N-NH2), -R b -OR a , -R b -OC(O)-R a , -R b -OC(O)-OR a , -R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)Ra , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (wherein t is 1 or 2), -R b -S(O) t R a (wherein t is 1 or 2), -R b -S(O) t OR a (wherein t is 1 or 2) and -R b -S(O) t N(R a )2 (wherein t is 1 or 2), and each R a is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl; and each R a is, if allowed by the valence, alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO2), imino (=NH), oximo (=N-OH), hydrazine (=N-NH2), -R b -OR a , -R b -OC(O)-R a , -R b -OC(O)-OR a , -R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)Ra , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (wherein t is 1 or 2), -R b -S(O) t R a (wherein t is 1 or 2), -R b -S(O) t OR a (wherein t is 1 or 2) and -R b -S(O) t N(R a )2 (wherein t is 1 or 2), and each R b are independently selected from a direct bond or a straight or branched alkylene, alkenylene, or alkynylene chain; and each R c is a straight or branched alkylene, alkenylene or alkynylene chain.

[0137] As used herein, the term "radical of a compound" refers to a structure derived from a parent compound by removing one or more atoms, e.g., hydrogen atoms. In one embodiment, a "radical of a compound" is a monovalent radical derived from removing one hydrogen atom from a parent compound.

[0138] It should be understood that a particular radical naming convention can include either a mono-radical or a di-radical, depending on the context. For example, if a substituent requires two points of attachment to the remainder of the molecule, the substituent is understood to be a di-radical. For example, a substituent identified as alkyl, which requires two points of attachment, includes di-radicals such as -CH-, -CHCH-, -CHCH(CH)CH-, etc. Other radical naming conventions explicitly indicate that the radical is a di-radical, such as "alkylene," "alkenylene," "arylene," etc.

[0139] Whenever a substituent is depicted as a di-radical (i.e., having two points of attachment to the rest of the molecule), it is to be understood that the substituent may be attached in any directional configuration unless otherwise indicated.

[0140] "Tautomer" refers to a molecule capable of proton transfer from one atom of the molecule to another atom of the same molecule. The compounds presented herein, in some embodiments, exist as tautomers. In situations where tautomerization is possible, a chemical equilibrium of tautomers exists. The exact ratio of tautomers depends on several factors, including physical conditions, temperature, solvent, and pH. Some examples of tautomer equilibrium include:

[0141] [ka] Examples include:

[0142] As used herein, the term "unsubstituted" may mean that the specified group bears no substituents other than the listed moieties (e.g., where a valency is filled by hydrogen).

[0143] "Stable compound" and "stable structure" can refer to a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0144] As used herein, the phrases "parenteral administration" and "parenterally administered" refer to modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal infusion and injection.

[0145] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms that are within the scope of sound medical judgment and are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0146] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and (10) soybean oil; (11) glycols, such as propylene glycol; (12) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (13) esters, such as ethyl oleate and ethyl laurate; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic, compatible substances used in pharmaceutical formulations.

[0147] The term "carrier" as used in this disclosure can encompass carriers, excipients, and diluents, and can refer to a material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, that is involved in carrying or transporting a pharmaceutical agent, such as one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates (e.g., hydrates), isomers (e.g., stereoisomers), and tautomers thereof, from one organ or body part of a subject to another. The carrier should be selected based on the compatibility and release profile properties of the desired dosage form. Exemplary carrier materials can include, for example, adjuvants, binders, suspending agents, disintegration agents, bulking agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, spray-dried dispersions, and the like. See, e.g., Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1975. Exemplary carrier materials may also include, but are not limited to, any adjuvant, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier approved by the U.S. Food and Drug Administration as acceptable for use in humans or veterinary medicine.

[0148] The terms "pharmaceutically acceptable" or "pharmacologically acceptable" can refer to a material that is biologically or otherwise undesirable, which may be administered to an individual without causing any substantial undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0149] A "pharmaceutical composition" may refer to a formulation of a compound of the present disclosure and a biologically active compound in a vehicle generally accepted in the art for delivering the compound to a subject, e.g., a mammal or a human. Such a vehicle may include any pharmaceutically acceptable carrier therefor.

[0150] The terms "subject," "individual," and "patient" may be used interchangeably and may refer to humans and non-human mammals (e.g., non-human primates, dogs, horses, cats, pigs, cows, ungulates, rabbits, etc.). In various embodiments, a subject may be a human (e.g., an adult male, adult female, adolescent male, adolescent female, boy, girl) under the care of a physician or other healthcare professional, as an outpatient in a hospital or in other clinical settings. In certain embodiments, a subject may not be under the care of a physician or other healthcare professional or on medication.

[0151] As used herein, the phrase "subject in need thereof" refers to a subject afflicted with or at risk of a condition that is treated prophylactically or therapeutically with a compound or salt described herein, as described below.

[0152] As used herein, "agent" or "biologically active agent" refers to a biological compound, pharmaceutical, or chemical compound or other moiety. Non-limiting examples include organic or inorganic molecules, peptides, proteins, peptide nucleic acids (PNAs), oligonucleotides (including, e.g., aptamers and polynucleotides), antibodies, antibody derivatives, antibody fragments, vitamin derivatives, carbohydrates, toxins, branched-chain amino acids in free amino acid form or their metabolites, or chemotherapeutic compounds. A variety of compounds can be synthesized, for example, small molecules and oligomers (e.g., oligopeptides and oligonucleotides), as well as synthetic organic compounds based on various core structures. Additionally, compounds for screening can be obtained from a variety of natural sources, such as plant or animal extracts. One skilled in the art can readily appreciate that there is no limit as to the structural nature of the agent.

[0153] The terms "administer," "administered," "administers," and "administering" are defined as providing a composition to a subject via a route known in the art, including, but not limited to, intravenous, intraarterial, oral, parenteral, buccal, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal, or intraperitoneal routes of administration. In some embodiments, oral administration of a composition may be used. The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or salt described herein sufficient to accomplish the intended use, including, but not limited to, the treatment of diseases defined below. A therapeutically effective amount may vary depending on the intended use (in vitro or in vivo) or the subject and condition being treated, e.g., the subject's weight and age, the severity of the condition, the method of administration, etc., and can be readily determined by one of ordinary skill in the art. The term can also apply to a dose capable of inducing a specific response in target cells, e.g., reduced proliferation or down-regulation of the activity of a target protein. Specific doses may vary depending on the particular compound selected, the dosing regimen followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the bodily delivery system to which the compound is delivered.

[0154] As used herein, "treatment" or "treating" refers to an approach for obtaining a beneficial or desired result with respect to a disease, disorder, or medical condition, including, but not limited to, a therapeutic benefit and / or a preventative benefit. In certain embodiments, treating or treating involves administering a compound or composition disclosed herein to a subject. A therapeutic benefit can include eradication or amelioration of the underlying disorder being treated. A therapeutic benefit can also be achieved by eradicating or ameliorating one or more physiological symptoms associated with the underlying disorder, e.g., observing an improvement in the subject, even though the subject may still be afflicted with the underlying disorder. In certain embodiments, for preventative benefit, a composition is administered to a subject at risk of developing a particular disease or reporting one or more physiological symptoms of a disease, even though a diagnosis of the disease may not have been made. Treating can include, for example, reducing, delaying, or alleviating the severity of one or more symptoms of a disease or condition, or it can include reducing the frequency with which a patient experiences symptoms of a disease, deficiency, disorder, or adverse condition. Treating may be used herein to refer to a method that results in some degree of treatment or amelioration of a disease or condition, and various outcomes toward that end may be envisioned, including, but not limited to, complete prevention of the condition.

[0155] In certain embodiments, the term "prevent" or "preventing" in relation to a disease or disorder may refer to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in a treated sample compared to an untreated control sample, or delays the onset of, or reduces the severity of, one or more symptoms of the disorder or condition compared to an untreated control sample.

[0156] The term "prodrug" is intended to encompass compounds that are converted under physiological conditions into therapeutically pharmaceutical agents of the present disclosure. A common method for making prodrugs is to include one or more selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by enzymatic activity of the host animal. For example, esters or carbonates (e.g., esters of alcohols or carboxylic acids or esters of carbonates and phosphonates) are preferred prodrugs of the present disclosure.

[0157] As used herein, the terms "treat," "treating," or "treatment" can include alleviating, reducing, or relieving the symptoms of a disease or condition, preventing additional symptoms, relieving or preventing the underlying cause of a symptom, inhibiting a disease or condition, for example, halting the onset of a disease or condition, relieving a disease or condition, causing regression of a disease or condition, alleviating conditions caused by a disease or condition, or prophylactically and / or therapeutically stopping the symptoms of a disease or condition.

[0158] The terms "effective amount" or "therapeutically effective amount," when used in connection with one or more compounds or pharmaceutical compositions, can refer to a quantity of one or more compounds or pharmaceutical compositions sufficient to bring about a desired biological result. This result can be a reduction and / or alleviation of the signs, symptoms, or causes of a disorder, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic use can be the amount of a pharmaceutical composition comprising one or more compounds disclosed herein, or pharmaceutically acceptable salts, solvates (e.g., hydrates), isomers (e.g., stereoisomers), and tautomers thereof, required to bring about a clinically significant reduction in a disorder. An appropriate "effective amount" in any individual case can be determined by one of ordinary skill in the art using routine experimentation.

[0159] In certain embodiments, the terms "disease" and "condition" may be used interchangeably or may differ in that a particular illness or condition may not yet be recognized as a disease because it may not have a known causative agent (and therefore its etiology is not yet understood), but rather only as an undesirable state or syndrome in which a more or less specific set of symptoms has been identified by a clinician.

[0160] The terms "antibacterial agent," "antibiotic," or "antibacterial compound" are used interchangeably herein and may refer to agents or compounds with either bactericidal or bacteriostatic activity. The term "inhibiting growth" may refer to slowing the rate at which a particular bacterial population increases in number. Thus, the term may include situations in which a bacterial population increases, but at a reduced rate, as well as situations in which the growth of the population is stopped, and situations in which the number of bacteria in a population is reduced or even eliminated. The activity of an antibacterial agent or compound is not necessarily limited to bacteria, but may also include general antimicrobial activity against parasites or fungi, or general antiviral activity against viruses.

[0161] Compounds of the Disclosure In general, the present disclosure provides aminoglycoside compounds that can be characterized as having antibacterial activity.

[0162] In one embodiment, formula (I)

[0163] [ka] (In the formula, R a is hydrogen (H) or methyl, R b is C 1~6 alkyl, or of formula (II)

[0164] [ka] The structure is Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR 4a R 4b and R 4a and R 4b is H and C 1~3 alkyl, or R 1 and R 4 together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N), or a pharmaceutically acceptable salt or solvate thereof.

[0165] In another embodiment, the compound of formula (III)

[0166] [ka] (In the formula, R a is H or methyl, R b is H, C 1~6 alkyl, or of formula (II)

[0167] [ka] The structure is Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR 4a R 4b and R 4a and R 4b is hydrogen and C 1~3 alkyl, or R 1 and R 4 together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N), or a pharmaceutically acceptable salt or solvate thereof.

[0168] Those skilled in the art will appreciate that the compounds of the present disclosure according to formula (I) and formula (III) each share a common hexahydropyranyl ring system.

[0169] [ka] (wherein Z is H (formula (I)) or methyl (formula (III))).

[0170] Without wishing to be bound by any particular theory, it is believed that the particular hexahydropyranyl ring system of compounds according to Formula (I) and Formula (III) may impart advantageous antibacterial activity to compounds containing such ring systems. Compounds of Formula (I) and Formula (III) may further contain additional, optionally substituted, saturated carbocyclic or heterocyclic ring systems that together comprise an aminoglycoside nucleus (e.g., tricyclic systems, such that compounds of Formula (I) and Formula (III) each contain four connected rings, including the hexahydropyranyl ring shown). Compounds of Formula (I) and Formula (III) may sometimes be referred to herein as "ABCD compounds" in reference to the tetracyclic (aminoglycoside) containing compounds, where the A, B, C, and D rings are as identified below.

[0171] [ka] For such ring systems, conventional numbering is applied herein. For simplicity, the conventional numbering is shown below with reference to a simplified, non-limiting ABCD ring scaffold (ABCD', i.e., with many omitted substituents compared to formulas (I) and (III) for ease of visualization).

[0172] [ka] The structure of the ABCD' ring scaffold can alternatively be represented in a three-dimensional format as shown below.

[0173] [ka] In some embodiments, Formula (I)

[0174] [ka] [In the formula, R a is hydrogen (H) or methyl, R b is C 1~6 alkyl, or of formula (II)

[0175] [ka] It has a structure according to Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR4a R 4b and R 4a and R 4b is H and C 1~3 alkyl, or R 1 and R 4 together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N.

[0176] In some embodiments, R a is H.

[0177] In some embodiments, R a is methyl (CH3).

[0178] In some embodiments, R b is C 1~6 Alkyl, for example methyl, ethyl, propyl, butyl (including isobutyl and sec-butyl), pentyl (including all its potential isomers) or hexyl (including all its potential isomers).

[0179] In some embodiments, R b is ethyl (C2H5).

[0180] In some embodiments, R b is ethyl, and the compound of formula (I) has the structure

[0181] [ka] It has.

[0182] In some embodiments, R b is ethyl, and the compound of formula (I) has the structure

[0183] [ka] It has.

[0184] In some embodiments, R b has a structure according to formula (II) as defined herein above.

[0185] In some embodiments, with respect to Formula (II), Q is NH, n is an integer from 1 to 3, and R 1 is hydrogen and R 2 and R 3 each occurrence is independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H. In some embodiments, n is 2.

[0186] In some embodiments, R b teeth

[0187] [ka] and n is 1, 2, or 3.

[0188] In some embodiments, R b teeth,

[0189] [ka] is selected from the group consisting of:

[0190] In some embodiments, R b teeth,

[0191] [ka] is selected from the group consisting of:

[0192] In some embodiments, the compound of formula (I) is

[0193] [ka] has a structure selected from the group consisting of:

[0194] In some embodiments, the compound of formula (I) has the structure

[0195] [ka] It has.

[0196] In some embodiments, the compound of formula (I) has the structure

[0197] [ka] It has.

[0198] In some embodiments, the compound of formula (I) is

[0199] [ka] has a structure selected from the group consisting of:

[0200] In some embodiments, the compound of formula (I) has the structure

[0201] [ka] It has.

[0202] In some embodiments, the compound of formula (I) has the structure

[0203] [ka] It has.

[0204] In another embodiment, the compound of formula (III)

[0205] [ka] [In the formula, R ais H or methyl, R b is H, C 1~6 alkyl, or of formula (II)

[0206] [ka] The structure is Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR 4a R 4b and R 4a and R 4b is hydrogen and C 1~3 alkyl, or R 1 and R 4 together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N.

[0207] In some embodiments, R a is H.

[0208] In some embodiments, R a is methyl.

[0209] In some embodiments, the compound of formula (III) is

[0210] [ka] has a structure selected from the group consisting of:

[0211] In some embodiments, R b is H.

[0212] In some embodiments, R b is C 1~6 alkyl, such as methyl, ethyl, propyl, butyl (including isobutyl and sec-butyl), pentyl (including all its potential isomers), or hexyl (including all its potential isomers). In some embodiments, R b is ethyl.

[0213] In some embodiments, R b is ethyl and the compound of formula (I) is

[0214] [ka] has a structure selected from the group consisting of:

[0215] In some embodiments, R b has a structure according to formula (II) as defined herein above.

[0216] In some embodiments, with respect to Formula (II), Q is NH, n is an integer from 1 to 3, and R 1 is hydrogen and R 2 and R 3 each occurrence is independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H. In some embodiments, n is 2.

[0217] In some embodiments, R b teeth,

[0218] [ka] and n is 1, 2, or 3.

[0219] In some embodiments, R b teeth,

[0220] [ka] is selected from the group consisting of:

[0221] In some embodiments, R b teeth,

[0222] [ka] is selected from the group consisting of:

[0223] In some embodiments, the compound of formula (III) is

[0224] [ka] has a structure selected from the group consisting of:

[0225] In some embodiments, the compound of formula (III) has the structure

[0226] [ka] It has.

[0227] In some embodiments, the compound of formula (III) has the structure

[0228] [ka] It has.

[0229] In some embodiments, the compound of formula (III) has the structure

[0230] [ka] It has.

[0231] In some embodiments, the compound of formula (III) has the structure

[0232] [ka] It has.

[0233] In some embodiments, the compound of formula (III) has the structure

[0234] [ka] It has.

[0235] In some embodiments, the compound of formula (III) has the structure

[0236] [ka] It has.

[0237] In some embodiments, the compound of formula (III) has the structure

[0238] [ka] It has.

[0239] In some embodiments, the compound of formula (III) has the structure

[0240] [ka] It has.

[0241] Isotopes and isotopically labeled compounds The compounds described herein may exhibit natural isotopic abundance, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number but an atomic mass or mass number different from that predominantly found in nature. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure. Thus, a reference to an element is meant to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, that isotope is also included. For example, hydrogen is 1 H (protium), 2 H (deuterium) and 3 H (tritium). Protium is the most abundant hydrogen isotope in nature. Enrichment with deuterium may confer certain therapeutic advantages, such as increased half-life and / or exposure in vivo, or may result in compounds useful for investigating in vivo pathways of drug excretion and metabolism. Isotopically enriched compounds may be prepared by conventional techniques well known to those skilled in the art.

[0242] The compounds described herein further include all pharmaceutically acceptable isotopically labeled compounds. An "isotopically" or "radiolabeled" compound may be a compound in which one or more atoms have been replaced or substituted with an atom having an atomic mass or mass number that is different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). For example, in some embodiments, in the compounds described herein, a hydrogen atom has been replaced or substituted with one or more deuterium or tritium atoms.

[0243] Certain isotopically labeled compounds of the present disclosure, for example, those incorporating a radioactive isotope, may be useful in drug and / or substrate tissue distribution studies. The radioactive isotope tritium, i.e. 3 H and carbon-14, i.e. 14 C can be particularly useful for this purpose due to its ease of incorporation and ready means of detection. Heavier isotopes, such as deuterium, i.e.2 Substitution with H may confer certain therapeutic advantages due to greater metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances. In some embodiments, the compound comprises at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of the present disclosure can be replaced or substituted with deuterium. In some embodiments, the compound comprises two or more deuterium atoms. In some embodiments, the compound comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Suitable isotopes that may be incorporated into the compounds described herein are: 2 H (deuterium is also written as D), 3 H (tritium is also written as T), 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125 I and 131 Positron-emitting isotopes, including but not limited to I, 11 C. 18 F, 15 O and 13 Substitution with N may be useful for positron emission tomography (PET) studies.

[0244] Isotopically labeled versions of the compounds disclosed herein may generally be prepared by procedures analogous to those disclosed in the schemes and / or examples herein, substituting the appropriate isotopically labeled reagent for the non-isotopically labeled reagent.

[0245] The disclosure herein is also meant to encompass the in vivo metabolic products of the disclosed compounds. Such products may result, primarily through enzymatic processes, from, for example, oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound. Accordingly, the disclosure may include compounds produced by a process comprising administering a compound of the present disclosure to a subject, e.g., a mammal, for a period of time sufficient to yield a metabolic product thereof. Such products are typically identified by administering a radiolabeled compound of the present disclosure to a subject, e.g., a rat, mouse, guinea pig, monkey, or human, at a detectable dose, allowing sufficient time for metabolism to occur, and isolating the conversion product from urine, blood, or other biological sample.

[0246] Salts and solvates The present disclosure further provides pharmaceutically acceptable salts, solvates (e.g., hydrates), and any combination of the compounds disclosed herein. The use of the terms "salt," "hydrate," "solvate," and the like is intended to apply equally to salts, hydrates, or solvates of enantiomers, diastereoisomers, isomers, stereoisomers, rotamers, tautomers, positional isomers, or racemates of the disclosed compounds.

[0247] The term "salt" or "pharmaceutically acceptable salt" refers to salts derived from a variety of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts may be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium, and magnesium salts.In some embodiments, "pharmaceutically acceptable salts" include, for example, water soluble and water insoluble salts, such as acetate, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium salt, edetate calcium salt, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fiunarate, gluceptate, gluconate, glutamate, glycolyl arsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, sethionate, and the like. nate), lactate, lactobionate, laurate, magnesium salt, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate, 1,1-methene-bis-2-hydroxy-3-naphthoate, einbonate, pantothenate, phosphate / diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, diacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate salts.

[0248] The compounds of the present invention also include crystalline and amorphous forms of such compounds, pharmaceutically acceptable salts and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudomorphs, solvates, hydrates, non-solvated polymorphs (including anhydrates), conformational polymorphs and amorphous forms of the compounds, and mixtures thereof.

[0249] The compounds of the present disclosure, including their stereoisomers and tautomers, as well as any salts thereof, can exist as solvates. Crystallization often produces solvates of the compounds of the present disclosure. As used herein, the term "solvate" can refer to an aggregate containing one or more molecules of a compound of the present disclosure and one or more molecules of a solvent. The solvent can be water, in which case the solvate can be a hydrate. Alternatively, the solvent can be an organic solvent. Thus, the compounds and salts of the present disclosure can exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvated forms. While the compounds of the present disclosure can be true solvates, in other cases, the compounds of the present disclosure may merely contain incidental water or be a mixture of water and some incidental solvent.

[0250] Preparation of compounds Compounds of the present disclosure having an ABCD ring system as described herein above (e.g., compounds of Formula (I) and Formula (III)) can be prepared by methods known in the art of organic synthesis, as set forth in part by the synthetic schemes in the following description, examples, and figures, along with the guidance provided herein. In the schemes described and illustrated below, it is understood that protecting groups for sensitive or reactive groups may be employed as necessary in accordance with the general principles or chemistry provided herein. Protecting groups may be manipulated according to standard methods of organic synthesis (T.W. Greene and P.G.M.Wuts, "Protective Groups in Organic Synthesis," 3rd ed., Wiley, New York, 1999). These groups may be removed at a convenient stage of the compound synthesis using methods readily apparent to those skilled in the art based on the detailed teachings provided herein. The selected processes, as well as reaction conditions and order of execution, shall be consistent with this disclosure.

[0251] Generally, methods for preparing compounds of the present disclosure having an ABCD ring system involve a combination of reactions and conditions that can yield specific novel intermediate compounds. General Scheme 1 shows a representative, non-limiting strategy for preparing compounds having an ABCD ring system (e.g., compounds of Formula (I) and Formula (III)).

[0252] [ka] Each aspect of the method of preparation according to Scheme I is further described herein below. The reactions in Scheme 1 can be carried out through chemical reactions using standard synthetic chemistry procedures and practices, optionally using catalysts including, but not limited to, reagents such as Pd reagents, chiral ligands, and enzymes.

[0253] Preparation of A-ring compounds An exemplary preparation of compounds having an ABCD ring system of the present disclosure (e.g., compounds of Formula (I) and Formula (III)) according to Scheme 1 utilizes an A-ring compound. In some embodiments, the A-ring compound has the general structure (A):

[0254] [ka] (In the formula, X 1 is an amine, an amine precursor or a protected amino group; X 2 is an amine, an amine precursor or a protected amino group; R 5 is hydrogen (H) or methyl (CH).

[0255] In some embodiments, R 5 is H.

[0256] In some embodiments, R 5 is CH3.

[0257] In some embodiments, X 1is azide (N3). In some embodiments, X 1 is -N(Bn)(Cbz). In some embodiments, X 1 is -N(CH3)(Cbz).

[0258] In some embodiments, X 2 is an azide (N3).

[0259] In some embodiments, the A-ring compound has formula A1 (i.e., ring A1):

[0260] [ka] It has the following structure.

[0261] An exemplary scheme for preparing Ring A1 is shown in Figure 1 and described in more detail herein. Generally, Ring A1 can be prepared starting from 2-formyl-3,4-dihydro-2H-pyran using a combination of condensation, reduction, oxidation, alkylation, acylation, deprotection, and substitution reactions.

[0262] With reference to FIG. 1 , in some embodiments, 2-formyl-3,4-dihydro-2H-pyran is converted to a chiral sulfinyl imine with, for example, (S)-2-methylpropane-2-sulfinamide in the presence of a Lewis acid dehydrating agent, such as copper sulfate. The sulfinyl imine can then be reduced to the corresponding sulfinamide with a suitable reducing agent, such as sodium borohydride. In some embodiments, the resulting sulfinamide nitrogen is then protected with a suitable protecting group, such as benzyl. In some embodiments, the tetrahydropyran is converted to the corresponding halolactol by reaction with, for example, bromine or iodine under aqueous conditions. In some embodiments, the hydroxy group of the halolactol is oxidized using a suitable oxidizing agent, such as a hexavalent chromium-based reagent, to form the corresponding lactone. In some embodiments, an azide group is introduced by displacement of a halo (e.g., iodo) group with an azide ion source. In some embodiments, the lactone carbonyl is reduced with a suitable reducing agent, such as an aluminum hydride reagent, to form the corresponding lactol. The sulfinamide chiral auxiliary can then be cleaved, for example under acidic conditions, and the resulting amino group protected with a suitable protecting group such as benzyl carbamate to give ring A1.

[0263] In some embodiments, the A-ring compound has formula A2 (i.e., ring A2):

[0264] [ka] It has the following structure.

[0265] An exemplary scheme for preparing Ring A2 is shown in Figure 2. Referring to Figure 2, in some embodiments, an aldehyde is converted to a chiral sulfinyl imine, for example, with (S)-2-methylpropane-2-sulfinamide in the presence of a Lewis acid dehydrating agent, for example, copper sulfate. The sulfinyl imine is then treated with a suitable organometallic reagent, for example, magnesium methyl bromide, to provide a chiral sulfinamide. In some embodiments, the resulting sulfinamide nitrogen is then protected with a suitable protecting group, for example, benzyl. In some embodiments, the tetrahydropyran is converted to the corresponding halolactol by reaction with, for example, bromine or iodine under aqueous conditions. In some embodiments, the hydroxy group of the halolactol is oxidized using a suitable oxidizing agent, for example, a hexavalent chromium-based reagent, to form the corresponding lactone. In some embodiments, an azide group is introduced by displacement of a halo (e.g., iodo) group with an azide ion source. In some embodiments, the lactone carbonyl is reduced with a suitable reducing agent, for example, an aluminum hydride reagent, to form the corresponding lactol. The sulfinamide chiral auxiliary can then be cleaved, for example under acidic conditions, and the resulting amino group protected with a suitable protecting group such as benzyl carbamate to give ring A2.

[0266] In some embodiments, the A-ring compound has formula A3 (i.e., ring A3):

[0267] [ka] It has the following structure.

[0268] Ring A3 is described with respect to FIG. 2 but can be prepared according to the general scheme shown in FIG. 3 using the R-enantiomer of 2-methylpropane-2-sulfinamide.

[0269] In some embodiments, the A-ring compound has formula A4 (ring A4):

[0270] [ka] It has the following structure.

[0271] In some embodiments, the A-ring compound has formula A5 (ring A5):

[0272] [ka] It has the following structure.

[0273] Rings A4 and A5 can be prepared according to the general scheme for rings A2 and A3, respectively, as described with respect to Figures 2 and 3, replacing the benzyl protecting group with methyl.

[0274] Preparation of B-ring compounds Preparation of compounds having an ABCD ring system of the present disclosure (e.g., compounds of Formula (I) and Formula (III)) according to Scheme 1 utilizes a B-ring compound. In some embodiments, the B-ring compound has the general structure

[0275] [ka] (In the formula, X is an amine, an amine precursor, or a protected amino group; R is a protecting group.

[0276] In some embodiments, X is NH(Cbz) and R is Ac. Thus, in some embodiments, the B-ring compound has the formula B1 (i.e., ring B1):

[0277] [ka] It has the following structure.

[0278] An exemplary method for preparing Ring B1 is shown in Figure 4. Referring to Figure 4, generally, Ring B1 can be prepared starting from dihydrostreptomycin by hydrolysis of the guanidine group, N-protection, O-protection, and hydrolysis of the glycosyl linkage. Procedures for preparing such compounds are reported, for example, in International Patent Application Publications 2019194858 and 2019046126, both to Achaogen, Inc., each of which is incorporated herein by reference for its teachings regarding the synthesis of Ring B1.

[0279] Preparation of AB-ring compounds An exemplary preparation of compounds having an ABCD ring system of the present disclosure (e.g., compounds of Formula (I) and Formula (III)) according to Scheme 1 utilizes an AB-ring compound. Generally, with reference to Scheme 1, the A-ring and B-ring compounds are joined via a glycosidic linkage to form the AB-ring compound. Generally, the reaction is carried out under conditions that allow for chemical glycosylation by coupling a glycosyl donor and a glycosyl acceptor. For example, a hydroxyl group on the B-ring compound can serve as a glycosyl donor, which reacts with an A-ring compound having a suitable leaving group and acting as a glycosyl acceptor.

[0280] As a non-limiting example, the anomeric hydroxyl group of the A-ring compound can be converted to a suitable leaving group capable of reacting with a hydroxyl group on the B-ring compound to form an interglycosidic linkage, which can then react with a free hydroxyl group on the B-ring compound in a suitable solvent to form said linkage. Examples of suitable leaving groups include, but are not limited to, halides, thioalkyl groups, thioaryl groups, imidates, acetates, phosphates, O-pentenyl groups, mesylates, tosylates, and the like. In one embodiment, the A-ring compound is treated with an activating agent, such as trichloroacetonitrile, to form an imidate, and the B-ring compound is added, followed by the addition of an additional activating agent, such as a Lewis acid. In some embodiments, the Lewis acid is trimethylsilyl triflate, trimethylaluminum, or boron trifluoride etherate.

[0281] The structure of the AB-ring compound can vary. In some embodiments, the AB-ring compound has the general structure

[0282] [ka] (In the formula, X 1 , X 2 and X 3 are each independently selected from an amine, an amine precursor, and a protected amino group; R 5 is H or CH3, R 6 and R 7 are each independently selected from H, Bn, and Piv.

[0283] In some embodiments, R 5 is H.

[0284] In some embodiments, R 5 is CH3.

[0285] In some embodiments, X 1 is azide (N3). In some embodiments, X 1 is —NH(Cbz). In some embodiments, X 1 is -N(Bn)(Cbz). In some embodiments, X 1 is -N(CH3)(Cbz).

[0286] In some embodiments, X 2 is azide (N3). In some embodiments, X 2 is -NH(Cbz).

[0287] In some embodiments, X 3 is azide (N3). In some embodiments, X 3 is -NH(Cbz).

[0288] In some embodiments, R 6 is benzyl. In some embodiments, R 6 is pivaloyl.

[0289] In some embodiments, R 7 is H.

[0290] In some embodiments, the AB-ring compound is

[0291] [ka] has a structure selected from the group consisting of:

[0292] In some embodiments, the AB-ring compound is ring AB1. An exemplary scheme for preparing ring AB1 is shown in Figure 5. In one embodiment, with reference to Figure 5, ring A1 is activated by treatment with an activating reagent such as, for example, trichloroacetonitrile or 2,2,2-trifluoro-N-phenyl-acetimidoyl chloride, followed by addition of ring B1 and a Lewis acid, such as bismuth triflate, to give a glycosylated product, from which the acetate protecting group is then removed with, for example, sodium methoxide or methanol in the presence of a carbonate base. The resulting 1,2-diol functionality is then protected, for example, by conversion to a ketal. One suitable ketal is formed from the reaction of a diol with cyclohexanone or cyclohexanone dimethyl ketal. The remaining B-ring hydroxy group can then be protected, for example, as a benzyl ether. The ketal protecting group can then be removed to give the 1,2-diol. To give the desired single free hydroxyl group of ring AB1, adjacent amino and hydroxyl groups are protected. One suitable protection strategy is the incorporation of amino and hydroxyl groups into the oxazolidinone. One suitable means of forming the oxazolidinone is treatment with a strong non-nucleophilic base, such as sodium hydride, which eliminates the benzyl alcohol to form the oxazolidinone.

[0293] In some embodiments, the AB-ring compound is ring AB2. An exemplary scheme for preparing ring AB2 is shown in Figure 6. The scheme utilizes the same general strategy as shown in Figure 5, except that ring A1 is replaced with ring A2.

[0294] In some embodiments, the AB-ring compound is ring AB3. An exemplary scheme for preparing ring AB3 is shown in Figure 7. The scheme utilizes the same general strategy as shown in Figure 5, replacing ring A1 with ring A3.

[0295] Additional AB ring compounds can be prepared similarly, for example, using alternative A ring compounds (e.g., ring A4 or ring A5), alternative B ring compounds, or both, and varying protecting group strategies as necessary.

[0296] Preparation of CD-ring compounds An exemplary preparation of compounds having an ABCD ring system of the present disclosure (e.g., compounds of Formula (I) and Formula (III)) according to Scheme 1 utilizes a CD-ring compound. In some embodiments, the CD-ring compound is a compound having a ring CD1.

[0297] [ka] is. Ring CD1 can be prepared, for example, according to the procedures reported in International Patent Application Publication No. 2019 / 194858 to Achaogen, Inc., which is incorporated herein by reference for the preparation of CD-ring compounds.

[0298] In some embodiments, the CD-ring compound is a ring CD2

[0299] [ka] is. Ring CD2 can be prepared from neomycin B, for example, according to the procedure reported in International Patent Application Publication No. 2019 / 194858 to Achaogen, Inc., which is incorporated herein by reference for the preparation of CD-ring compounds.

[0300] Preparation of ABCD ring compounds Preparation of compounds having an ABCD ring system according to Scheme 1 (i.e., compounds of Formula (I) and Formula (III)) generally involves connecting AB-ring and CD-ring compounds via a glycosidic linkage, each as described herein above with respect to Scheme 1. As a non-limiting example, the anomeric hydroxyl group of the CD-ring compound is converted into a suitable leaving group (acceptor) capable of reacting with a hydroxyl group (donor) on the AB-ring compound to form an interglycosidic linkage, which then reacts with a free hydroxyl group on the AB-ring compound in a suitable solvent to form said linkage. Examples of suitable leaving groups include, but are not limited to, halides, thioalkyl groups, thioaryl groups, imidates, acetates, phosphates, O-pentenyl groups, mesylates, tosylates, and the like. In one embodiment, the CD-ring compound is treated with an activating agent, e.g., trichloroacetonitrile, to form an imidate, and the AB-ring compound is added, followed by the addition of an additional activating agent, e.g., a Lewis acid. In some embodiments, the Lewis acid is trimethylsilyl triflate, trimethylaluminum, or boron trifluoride etherate.

[0301] In some embodiments, the AB-ring compound is ring AB1, ring AB2, or ring AB3. In some embodiments, the CD-ring compound is ring CD1. Exemplary schemes for preparing ring ABCD1, ring ABCD2, and ring ABCD3 are shown in Figures 8, 9, and 10, respectively.

[0302] In one embodiment, Ring ABCD1 (i.e., a precursor to a compound of Formula (I)) can be prepared according to the scheme shown in Figure 8. With reference to Figure 8, Ring CD1 is activated by treatment with an activating agent, such as trichloroacetonitrile, followed by the addition of Ring AB1 and a Lewis acid to provide a compound having an ABCD ring system. In some embodiments, the Lewis acid is boron trifluoride etherate. Following the glycosylation reaction, the protected amino and hydroxyl groups, attached as an oxazolidinone, are hydrolyzed under basic conditions to provide Ring ABCD1.

[0303] In one embodiment, Ring ABCD2 (i.e., a precursor to a compound of Formula (III)) can be prepared according to the scheme shown in Figure 9. With reference to Figure 9, Ring CD1 is activated, for example, by treatment with trichloroacetonitrile, followed by the addition of Ring AB2 and a Lewis acid to provide a compound having an ABCD ring system. In some embodiments, the Lewis acid is boron trifluoride etherate. Following the glycosylation reaction, the protected amino and hydroxyl groups, attached as an oxazolidinone, are hydrolyzed under basic conditions to provide Ring ABCD2.

[0304] In one embodiment, ring ABCD3 (i.e., the precursor to the compound of formula (III)) can be prepared according to the scheme shown in Figure 10. The scheme utilizes the same transformations described above with respect to Figure 9, except that ring AB2 is replaced with ring AB3.

[0305] Additionally, ABCD ring compounds can be prepared using schemes similar to those described herein above, substituting the appropriate AB-ring and CD-ring compounds while adjusting protecting group strategies accordingly. Those skilled in the art will understand the adjustments made to protecting group strategies to accommodate the various combinations of potential AB-ring and CD-ring compounds envisioned within the general formulae (e.g., (I) and (III)) disclosed herein.

[0306] Substituent R b Introduction of In any of the foregoing embodiments, the ABCD compound (i.e., a compound of formula (I) or formula (III)) may contain a substituent R b In some embodiments, R b is H, C 1~6 is alkyl or has a structure according to formula (II):

[0307] In some embodiments, the ABCD compound is a compound of formula (III) and has the substituent R b and R b is H. In such embodiments, the corresponding ABCD compound is subjected to global deprotection to provide a compound of formula (III). In some embodiments, global deprotection can be achieved, for example, by hydrogenolysis to cleave all benzyl protecting groups and reduce the azide to the corresponding amine. Remaining protecting groups (e.g., acetate) can be removed by acidic or alkaline hydrolysis either before or after hydrogenolysis, or can be cleaved spontaneously by methanolysis under hydrogenolysis conditions.

[0308] In some embodiments, the ABCD compound is a compound of Formula (I) or Formula (III) and has a substituent R b and R b is C 1~6 In such embodiments, the corresponding ABCD compound is subjected to reductive alkylation with the desired aldehyde and a suitable reducing agent (e.g., acetoxy or cyanoborohydride) to provide the corresponding N-alkyl compound. The resulting N-alkyl compound is then subjected to global deprotection to provide a compound of Formula (I) or Formula (III). In some embodiments, global deprotection can be achieved, for example, by hydrogenolysis to cleave all benzyl protecting groups and reduce the azide to the corresponding amine. Remaining protecting groups (e.g., acetate) can be removed by acidic or alkaline hydrolysis either before or after hydrogenolysis, or can be spontaneously cleaved by methanolysis under hydrogenolysis conditions.

[0309] In some embodiments, the ABCD compound is a compound of Formula (I) or Formula (III) and has a substituent R b and R b is represented by formula (II)

[0310] [ka] (In the formula, Q is NH, O or S; n is an integer from 0 to 4, R 1 is hydrogen or C 1~3 is alkyl, R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H, C 1~3 Alkyl or -C(=NH)NR 4a R 4b and R 4a and R 4b is hydrogen and C 1~3 alkyl, or R 1 and R 4 together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N).

[0311] Thus, the preparation of ABCD compounds of formula (I) or formula (III) can, in various embodiments, be carried out by reacting R b is reacted with the free NH group of the corresponding ABCD ring compound to form the substituent R c which, after additional steps as indicated, can be used to obtain an ABCD ring compound having the desired substituent R b An ABCD ring compound of formula (I) or (III) having the formula:

[0312] Referring to the structure of formula (II), in some embodiments, Q is NH and R 1 is hydrogen and R 2and R 3 each occurrence is independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R 4 is H. Thus, in some embodiments, the substituent R c is represented by formula (IIa)

[0313] [ka] (In the formula, Pg is a protecting group, Z is an amine precursor or a protected amino group; The wavy line indicates the point of attachment to the amine on the B-ring.

[0314] In some embodiments, Z is azide (N3). In some embodiments, Z is NHCBz.

[0315] In some embodiments, Pg is benzyl. In some embodiments, Pg is para-methoxybenzyl (PMB).

[0316] In the foregoing embodiment, one skilled in the art would recognize that the substituent R c In general, a suitable reactant for introducing the substituent R is identified, such as the corresponding carboxylic acid or an activated version thereof (i.e., the wavy line in the compound of formula (IIa) represents an OH group or a leaving group). c The introduction of R c Furthermore, one skilled in the art will appreciate that the final global deprotection (e.g., by hydrogenolysis) of the ABCD ring compound can be carried out by coupling the substituent R c and / or reduce the azide to an amine, and the corresponding substituent R of the final ABCD ring compound of formula (I) or formula (III) is removed. b Understand what you can get.

[0317] In some embodiments, R b teeth,

[0318] [ka] has a structure selected from the group consisting of: Thus, in some embodiments, the substituent R c has a structure according to formula (IIa), n is 2, and R 2 and R 3 is each occurrence independently selected from the group consisting of hydrogen and F. In such embodiments, the substituent R c teeth,

[0319] [ka] has a structure selected from the group consisting of:

[0320] In some embodiments, R b teeth,

[0321] [ka] is selected from the group consisting of R c teeth,

[0322] [ka] It is one of them.

[0323] In some embodiments, the substituent R b is the structure

[0324] [ka] and the corresponding reactant has the structure

[0325] [ka] The compound is (S)-2-(benzyloxy)-4-((2-oxo-2-phenyl-1λ-ethyl)amino)butanoic acid having the formula: Both the depicted reactant and the corresponding substituent Rb are referred to herein as "HABA," referring to the hydroxy-aminobutyric acid portion of the compound.

[0326] In some embodiments, the substituent R b is the structure

[0327] [ka] and the corresponding reactant has the structure

[0328] [ka] (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid having the formula: Both the depicted reactants and the corresponding substituents Rb are referred to herein as "F-HABA," in reference to the fluoro-hydroxy-aminobutyric acid portion of the compound.

[0329] In one embodiment, (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid (F-HABA) can be prepared according to the scheme shown in Figure 14. With reference to Figure 14, D-isoascorbic acid is protected as a ketal and oxidatively cleaved to give an α-hydroxy ester. The ester is then reduced to an alcohol, and the primary hydroxyl group is converted to an azide. The secondary hydroxyl group is then converted to a fluoro group in a stereochemical inversion. The ketal is removed, and the two hydroxyl groups are differentially protected. For example, the primary hydroxyl group is protected as a silyl ether and the secondary hydroxyl as a benzyl ether (e.g., para-methoxybenzyl). Cleavage of the silyl ether and oxidation of the primary hydroxymethyl group to the corresponding carboxylic acid then gives the desired (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid.

[0330] Pharmaceutical Formulations and Compositions The compounds described herein are contemplated as useful as pharmaceutical compositions for administration to a subject in need thereof. The pharmaceutical compositions may include at least a compound or salt described herein, as well as one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersing agents, suspending agents, and / or thickening agents.

[0331] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers, including excipients and auxiliary agents. Formulations can be modified depending on the selected route of administration. Pharmaceutical compositions containing the compounds or salts described herein can be prepared, for example, by lyophilizing the compounds or salts, mixing, dissolving, emulsifying, encapsulating, or encapsulating the compounds or salts. Pharmaceutical compositions can also contain the compounds or salts described herein in free base form or in pharmaceutically acceptable salt form.

[0332] The pharmaceutical compositions described herein can include at least one active ingredient (e.g., a compound or salt). The active ingredient can be encapsulated, for example, in microcapsules prepared by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin microcapsules and poly(methylmethacylate) microcapsules, respectively), colloidal drug-delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or macroemulsions.

[0333] The pharmaceutical compositions described herein can often further comprise more than one active compound (e.g., a compound or salt and other agents) as needed for the particular indication being treated. The active compounds can have complementary activities that do not adversely affect each other. For example, the composition can also include a chemotherapeutic agent, a cytotoxic agent, a cytokine, a growth inhibitory agent, an antihormonal agent, an antiangiogenic agent, and / or a cardioprotective agent. Such molecules can be present in combination in amounts effective for the intended purpose.

[0334] The compositions and formulations may be sterilized, which may be achieved by filtration through sterile filtration.

[0335] The compositions described herein can be formulated for administration as injections. Non-limiting examples of injection preparations can include sterile suspensions, solutions, or emulsions in oily or aqueous vehicles. Suitable oily vehicles can include, but are not limited to, lipophilic solvents or vehicles, such as fatty oils or synthetic fatty acid esters or liposomes. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension. The suspension can also contain suitable stabilizers. Injections can be formulated for bolus injection or continuous injection. Alternatively, the compositions described herein can be lyophilized or in powder form for reconstitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

[0336] For parenteral administration, the compound or salt can be formulated in a unit-dose injectable form (e.g., using a letter solution, suspension, or emulsion) with a pharmaceutically acceptable parenteral vehicle. Such a vehicle can be essentially non-toxic and non-therapeutic. The vehicle can be water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous vehicles, such as fixed oils and ethyl oleate, can also be used. Liposomes can be used as carriers. The vehicle can contain small amounts of additives, such as substances that improve isotonicity and chemical stability (e.g., buffers and preservatives).

[0337] Sustained-release preparations can also be prepared. Examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers that can contain the compound or salt, and these matrices can be in the form of shaped articles (e.g., films or microcapsules). Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactides, copolymers of L-glutamic acid and gamma-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPO™ (i.e., injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprorelide acetate), and poly-D-(-)-3-hydroxybutyric acid.

[0338] The pharmaceutical formulations described herein can be prepared for storage by mixing the compound or salt with a pharmaceutically acceptable carrier, excipient, and / or stabilizer. The formulation can be a lyophilized formulation or an aqueous solution. Acceptable carriers, excipients, and / or stabilizers can be non-toxic to the recipient at the dosages and concentrations used. Acceptable carriers, excipients, and / or stabilizers can include buffers, such as phosphate, citric acid, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives; polypeptides; proteins, such as serum albumin or gelatin; hydrophilic polymers; amino acids; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium; metal complexes; and / or nonionic surfactants or polyethylene glycol.

[0339] In some embodiments, the drug delivery composition may be incorporated into a system that includes a substrate that carries the composition to the site of administration or delivery or treatment. The substrate may remain with the composition after administration (or delivery of the composition) for a period of time or indefinitely, or may be removed after administration (or delivery of the composition), leaving the composition at the site of administration or delivery or treatment.

[0340] The compounds of formula (I) or formula (III) may be administered in their "natural" form or, if desired, in the form of salts, esters, amides, prodrugs, clathrates, derivatives, etc., provided that the salts, esters, amides, prodrugs, clathrates, or derivatives are pharmacologically suitable, e.g., effective in treating the pathology and / or its various symptoms, e.g., as described herein. Salts, esters, amides, clathrates, prodrugs, and other derivatives of the compounds of formula (I) or formula (III) are known to those skilled in the art of synthetic organic chemistry and may be prepared using standard procedures, e.g., as described by March (1992), Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 4th Edition, NY Wiley-Interscience, as described above.

[0341] For example, pharmaceutically acceptable salts can be prepared for any compound of Formula (I) or Formula (III) described herein having functional groups capable of forming salts. A pharmaceutically acceptable salt is any salt that retains the activity of the parent compound and does not impart any harmful or adverse effects to the subject and situation in which it is administered.

[0342] In various embodiments, pharmaceutically acceptable salts can be derived from organic or inorganic bases. Salts can be monovalent or polyvalent ions. Of particular interest are the inorganic ions lithium, sodium, potassium, calcium, and magnesium. Organic salts can be made with amines, particularly ammonium salts, such as mono-, di-, and trialkylamines or ethanolamines. Salts can also be formed with caffeine, tromethamine, and similar molecules.

[0343] Methods for formulating pharmaceutical compounds of Formula (I) or Formula (III) as salts, esters, amides, prodrugs, and the like are well known to those skilled in the art. For example, salts can be prepared from the free base using conventional methodology, typically involving reaction with a suitable acid. Generally, the base form of the drug is dissolved in a polar organic solvent, such as methanol or ethanol, and an acid is added thereto. The resulting salt precipitates or can be removed from solution by adding a less polar solvent. Suitable acids for preparing acid addition salts include, but are not limited to, both organic acids, such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like, as well as inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Acid addition salts can be reconverted to the free base by treatment with a suitable base. Particularly preferred acid addition salts of the compounds of formula (I) or formula (III) herein include halide salts, which may be prepared using, for example, hydrochloric acid or hydrobromic acid. Conversely, base salts of the compounds of formula (I) or formula (III) of the present disclosure are prepared in a similar manner using pharmaceutically acceptable bases, such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine, etc. Particularly preferred base salts include alkali metal salts, such as sodium and copper salts.

[0344] When preparing a salt form of a basic drug, the pKa of the counterion is preferably at least about 2 pH units lower than the pKa of the drug. Similarly, when preparing a salt form of an acidic drug, the pKa of the counterion is preferably at least about 2 pH units higher than the pKa of the drug. This allows the counterion to lower the pH of the solution below the pHmax, allowing the salt to reach a stable state where the solubility of the salt exceeds that of the free acid or base. The general rule of the difference in pKa units of the ionizable groups in the active pharmaceutical ingredient (API) and the acid or base means that proton transfer is energetically favorable. If the pKa of the API and the counterion are not significantly different, a solid complex may form but may rapidly become disequilibrium in an aqueous environment (i.e., decompose into individual entities of the drug and counterion).

[0345] Preferably, the counterion is a pharmaceutically acceptable counterion. Suitable anionic salt forms include acetate, benzenesulfonate, benzoate, benzilate, bicarbonate, bitartrate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycolyl arsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, and methyl bromide. Suitable cationic salt forms include, but are not limited to, methyl nitrate, methyl sulfate, mucinate, napsylate, nitrate, pamoate (embonate), pantothenate, phosphate and diphosphate, polygalacturonate, salicylate and disalicylate, stearate, monoacetate, succinate, sulfate, tannate, tartrate, teoclate, tosylate, triethiodide, valerate, etc., while suitable cationic salt forms include, but are not limited to, aluminum, benzathine, calcium, ethylenediamine, lysine, magnesium, meglumine, potassium, procaine, sodium, tromethamine, zinc, etc. Suitable cationic salt forms include, but are not limited to, benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, etc.

[0346] Preparation of esters typically involves functionalization of hydroxyl and / or carboxyl groups present within the molecular structure of a compound of Formula (I) or Formula (III). In certain embodiments, esters are typically acyl-substituted derivatives of free alcohol groups, i.e., moieties derived from carboxylic acids of the formula RCOOH, where R is alkyl, preferably lower alkyl. Esters can be reconverted to the free acids, if desired, by using conventional hydrogenolysis or hydrolysis procedures.

[0347] Amides may also be prepared using techniques known to those skilled in the art or described in the pertinent literature. For example, amides may be prepared from esters, using suitable amine reactants, or they may be prepared from anhydrides or acid chlorides by reaction with ammonia or a lower alkyl amine.

[0348] In various embodiments, the compounds of Formula (I) or Formula (III) described herein are useful for parenteral, topical, oral, nasal (or otherwise inhaled), rectal, or local administration, such as by aerosol or transdermal, for the prophylactic and / or therapeutic treatment of one or more pathologies / symptoms described herein (e.g., pathologies characterized by excessive amyloid plaque formation and / or deposition or unwanted amyloid or preamyloid processing).

[0349] The compounds of Formula (I) or Formula (III) described herein can also be combined with a pharmaceutically acceptable carrier (excipient) to form a pharmacological composition. The pharmaceutically acceptable carrier can contain one or more physiologically acceptable compounds that act, for example, to stabilize the composition or increase or decrease the absorption of the active agent. Physiologically acceptable compounds can include, for example, carbohydrates such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, protective and absorption enhancers such as lipids, compositions or excipients that decrease the clearance or hydrolysis of the active agent, or other stabilizers and / or buffers.

[0350] Other physiologically acceptable compounds, particularly for use in preparing tablets, capsules, gelcaps, and the like, include, but are not limited to, binders, diluents / fillers, disintegrants, lubricants, suspending agents, and the like.

[0351] In some embodiments, to produce oral dosage forms (e.g., tablets), excipients (e.g., lactose, sucrose, starch, mannitol, etc.), optional disintegrants (e.g., calcium carbonate, carboxymethylcellulose calcium, sodium starch glycollate, crospovidone, etc.), binders (e.g., pregelatinized starch, gum arabic, microcrystalline cellulose, carboxymethylcellulose, polyvinylpyrrolidone, hydroxypropylcellulose, cyclodextrin, etc.), and optional lubricants (e.g., talc, magnesium stearate, polyethylene glycol 6000, etc.) are added to, for example, the active ingredient(s) (e.g., a compound represented by a structure such as Formula I) and the resulting composition is compressed. Optionally, the compressed product is coated using known methods, for example, for taste masking or for intestinal dissolution or sustained release. Suitable coating materials include, but are not limited to, ethyl-cellulose, hydroxymethylcellulose, POLYOX® polyethylene glycol, cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, and Eudragit (Rohm & Haas, Germany; methacrylic-acrylic copolymer).

[0352] Other physiologically acceptable compounds include wetting agents, emulsifying agents, dispersing agents, or preservatives, which are particularly useful for preventing the growth or action of microorganisms. Various preservatives are well known and include, for example, phenol and ascorbic acid. One skilled in the art will recognize that the choice of a pharmaceutically acceptable carrier, including a physiologically acceptable compound, depends, for example, on the route of administration of the active agent and the specific physicochemical properties of the active agent. In certain embodiments, the excipients are sterile and generally free of undesirable matter. These compositions can be sterilized by conventional, well-known sterilization techniques. Sterility is not required for excipients of various oral dosage forms, such as tablets and capsules. USP / NF standards are usually sufficient.

[0353] Pharmaceutical compositions may be administered in a variety of unit dosage forms depending on the method of administration. Suitable unit dosage forms include, but are not limited to, powders, tablets, pills, capsules, lozenges, suppositories, patches, nasal sprays, injectibles, implantable sustained-release formulations, mucoadhesive films, topical varnishes, lipid complexes, etc.

[0354] Pharmaceutical compositions containing compounds of formula (I) or formula (III) can be prepared by conventional mixing, dissolving, granulating, dragee-making, mashing, emulsifying, encapsulating, entrapping or lyophilizing processes. Pharmaceutical compositions can be formulated in a conventional manner using one or more physiologically acceptable carriers, diluents, excipients or auxiliaries that facilitate the processing of the active agent into pharmaceutically usable preparations. The appropriate formulation depends on the selected route of administration.

[0355] In certain embodiments, the active agents described herein (e.g., compounds of Formula (I) or Formula (III) and compositions comprising the compounds) are formulated for oral administration. For oral administration, suitable formulations can be readily formulated by combining the active agent with pharmaceutically acceptable carriers suitable for oral delivery, as are well known in the art. Such carriers allow the active agents described herein to be formulated as tablets, pills, dragees, caplets, lozenges, gelcaps, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by the patient being treated. For oral solid formulations such as powders, capsules, and tablets, suitable excipients may include fillers such as sugars (e.g., lactose, sucrose, mannitol, and sorbitol), cellulose preparations (e.g., maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose), synthetic polymers (e.g., polyvinylpyrrolidone (PVP)), granulating agents, and binders. If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate, may be added. If desired, solid dosage forms may be sugar-coated or enteric-coated using standard techniques. The preparation of enteric-coated particles is disclosed, for example, in U.S. Pat. Nos. 4,786,505 and 4,853,230.

[0356] For administration by inhalation, the active agent is conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, capsules and cartridges of, for example, gelatin, for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0357] In various embodiments, the active agent can be formulated into a rectal or vaginal composition, such as a suppository or retention enema containing a conventional suppository base, such as cocoa butter or other glycerides. Methods of formulating active agents for rectal or vaginal delivery are well known to those skilled in the art (see Allen (2007) Suppositories, Pharmaceutical Press) and typically involve combining the active agent with a suitable base (e.g., a hydrophilic (PEG), a fat-soluble material, such as cocoa butter or Witepsol W45, an amphiphilic material, such as Suppocire AP, and polyglycolized glycerides). The base is selected and formulated for the desired melting / delivery profile.

[0358] For topical administration, the compounds of formula (I) or formula (III) may be formulated as solutions, gels, ointments, creams, suspensions, etc., as is well known in the art.

[0359] In certain embodiments, the compounds of Formula (I) or Formula (III) described herein are formulated for systemic administration (e.g., as an infusion) according to standard methods known to those skilled in the art. Systemic formulations include, but are not limited to, those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection, as well as those designed for transdermal, transmucosal, oral, or pulmonary administration. For injection, the active agents described herein can be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer, and / or in specific emulsion formulations. Solutions can contain formulating agents, such as suspending, stabilizing, and / or dispersing agents. In certain embodiments, the active agents are provided in powder form for reconstitution with a suitable vehicle, e.g., sterile, pyrogen-free water, prior to use. For transmucosal administration and / or the blood / brain barrier, penetrants appropriate to the barrier to be permeated can be used in the formulation. Such penetrants are generally known in the art. Injectable and inhalable formulations are generally obtained as sterile or substantially sterile formulations.

[0360] In addition to the formulations described above, the active agent can also be formulated as a depot preparation. Such long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. Thus, for example, the active agent can be formulated with a suitable polymeric or hydrophobic material (e.g., as an emulsion in an acceptable oil) or ion exchange resin, or as a sparingly soluble derivative, for example, as a sparingly soluble salt.

[0361] In certain embodiments, the active agents described herein can also be delivered through the skin using conventional transdermal drug delivery systems, i.e., transdermal "patches," in which the active agent is typically contained within a laminated structure that serves as a drug delivery device attached to the skin. In such structures, the drug composition is typically contained in a layer, or "reservoir," underlying an upper backing layer. It is recognized that the term "reservoir" in this context refers to the quantity of "active ingredient" available for ultimate delivery to the surface of the skin. Thus, for example, a "reservoir" can contain the active ingredient in an adhesive on the backing layer of a patch, or in any of a variety of different matrix formulations known to those skilled in the art. A patch can contain a single reservoir, or it can contain multiple reservoirs.

[0362] In one exemplary embodiment, the reservoir comprises a polymer matrix of a pharmaceutically acceptable contact adhesive material, which serves to attach the system to the skin during drug delivery. Examples of suitable skin contact adhesive materials include, but are not limited to, polyethylene, polysiloxane, polyisobutylene, polyacrylate, polyurethane, and the like. Alternatively, the drug-containing reservoir and the skin contact adhesive may exist as separate, distinct layers, with the adhesive underlying the reservoir, in which case it may be a polymer matrix as described above, or a liquid or hydrogel reservoir, or may take some other form. The backing layer in these laminates, which serves as the top surface of the device, preferably functions as the first component of the "patch" and provides much of its flexibility to the device. The material selected for the backing layer is preferably substantially impermeable to the active agent and any other materials present.

[0363] Alternatively, other pharmaceutical delivery systems can be used.For example, liposomes, emulsions, and microemulsions / nanoemulsions are well-known examples of delivery vehicles that can be used to protect and deliver pharmaceutically active compounds.Certain organic solvents, such as dimethyl sulfoxide, can also be used, but usually at the expense of greater toxicity.

[0364] In some embodiments, the compounds of Formula (I) or Formula (III) described herein are formulated into nanoemulsions. Nanoemulsions include, but are not limited to, oil-in-water (O / W) nanoemulsions and water-in-oil (W / O) nanoemulsions. Nanoemulsions may be defined as emulsions having an average droplet diameter ranging from about 20 to about 1000 nm. Typically, the average droplet size is about 20 nm or 50 nm to about 500 nm. The terms submicron emulsion (SME) and miniemulsion are used synonymously.

[0365] Exemplary oil-in-water (O / W) nanoemulsions include, but are not limited to, surfactant micelles, which are micelles composed of small molecule surfactants or detergents (e.g., SDS / PBS / 2-propanol); polymeric micelles, which are micelles composed of polymeric, copolymeric, or block copolymer surfactants (e.g., Pluronic L64 / PBS / 2-propanol); blended micelles, in which more than one surfactant component is present or in which one of the liquid phases (typically an alcohol or fatty acid compound) participates in the formation of the micelle (e.g., octanoic acid / PBS / EtOH); integrated micelles, which are blended micelles in which the active agent acts as a co-surfactant and forms an integral part of the micelle; and Pickering (solid phase) emulsions in which the active agent is associated with the exterior of solid nanoparticles (e.g., polystyrene nanoparticles / PBS / no oil phase).

[0366] Exemplary water-in-oil (W / O) nanoemulsions include, but are not limited to, surfactant micelles, which are micelles composed of small molecule surfactants or detergents (e.g., dioctyl sulfosuccinate / PBS / 2-propanol, isopropyl myristate / PBS / 2-propanol, etc.); polymeric micelles, which are micelles composed of polymeric, copolymeric, or block copolymer surfactants (e.g., PLURONIC® L121 / PBS / 2-propanol); blended micelles, in which more than one surfactant component is present or in which one of the liquid phases (typically an alcohol or fatty acid compound) participates in the formation of the micelle (e.g., capric / caprylic diglyceride / PBS / EtOH); integrated micelles, which are blended micelles in which the active agent serves as a co-surfactant and forms an integral part of the micelle (e.g., active agent / PBS / polypropylene glycol); and Pickering (solid phase) emulsions, in which the active agent is associated with the exterior of solid nanoparticles (e.g., chitosan nanoparticles / no aqueous phase / mineral oil).

[0367] As noted above, in some embodiments, the nanoemulsion includes one or more surfactants or detergents. In some embodiments, the surfactant is a non-anionic detergent (e.g., a polysorbate surfactant, a polyoxyethylene ether, etc.). Surfactants that find use in the present invention include, but are not limited to, surfactants such as the TWEEN®, TRITON®, and TYLOXAPOL® families of compounds.

[0368] In some embodiments, the emulsion further comprises one or more cationic halogen-containing compounds, including, but not limited to, cetylpyridinium chloride. In yet further embodiments, the composition further comprises one or more compounds that increase the interaction of the composition with the microorganism ("interaction enhancers") (e.g., a chelating agent such as ethylenediaminetetraacetic acid or ethylenebis(oxyethylenenitrilo)tetraacetic acid in a buffer).

[0369] In some embodiments, the nanoemulsion further comprises an emulsifier to aid in emulsion formation. Emulsifiers include compounds that aggregate at the oil / water interface to form a type of continuous membrane that prevents direct contact between two adjacent droplets. Certain embodiments of the present invention feature oil-in-water emulsion compositions that can be easily diluted with water to the desired concentration without compromising antipathogenicity.

[0370] In addition to discrete oil droplets dispersed in an aqueous phase, certain oil-in-water emulsions may also contain other lipid structures, such as small lipid vesicles (e.g., lipid spheres often consisting of several substantially concentric lipid bilayers separated from each other by layers of aqueous phase), micelles (e.g., amphiphilic molecules in small clusters of 50-200 molecules arranged with their polar head groups facing outward toward the aqueous phase and their nonpolar tails sequestered inward from the aqueous phase), or lamellar phases (lipid dispersions in which each particle consists of parallel amphiphilic bilayers separated by a thin film of water).

[0371] These lipid structures form as a result of hydrophobic forces that drive nonpolar residues (e.g., long hydrocarbon chains) away from water. The above lipid preparations can generally be described as surfactant lipid preparations (SLPs). SLPs are thought to be minimally toxic to mucous membranes and metabolized in the small intestine (see, e.g., Hamouda et al. (1998), J. Infect. Disease 180:1939).

[0372] In some embodiments, the emulsion comprises a discontinuous oil phase distributed in an aqueous phase, where the first component comprises an alcohol and / or glycerol, and the second component comprises a surfactant or halogen-containing compound. The aqueous phase can comprise any type of aqueous phase, including, but not limited to, water (e.g., deionized water, distilled water, tap water) and solutions (e.g., phosphate-buffered saline solutions or other buffer systems). The oil phase can comprise any type of oil, including, but not limited to, vegetable oils (e.g., soybean oil, avocado oil, linseed oil, palm oil, cottonseed oil, squalene oil, olive oil, canola oil, corn oil, rapeseed oil, safflower oil, and sunflower oil), animal oils (e.g., fish oil), flavor oils, water-insoluble vitamins, mineral oil, and motor oil. In some embodiments, the oil phase comprises 30-90% by volume of the oil-in-water emulsion (i.e., comprising 30-90% of the total volume of the final emulsion), more preferably 50-80%. Although the formulation need not be limited to a particular surfactant, in certain embodiments, the surfactant is a polysorbate surfactant (e.g., TWEEN 20®, TWEEN 40®, TWEEN 60®, and TWEEN 80®), phenoxypolyethoxyethanol (e.g., TRITON® X-100, X-301, X-165, X-102, and X-200, and TYLOXAPOL®), or sodium dodecyl sulfate.

[0373] In some embodiments, a halogen-containing component is present. The nature of the halogen-containing compound includes, in some preferred embodiments, chloride salts (e.g., NaCl, KCl, etc.), cetylpyridinium halides, cetyltrimethylammonium halides, cetyldimethylethylammonium halides, cetyldimethylbenzylammonium halides, cetyltributylphosphonium halides, dodecyltrimethylammonium halides, tetradecyltrimethylammonium halides, cetylpyridinium chloride, cetyltrimethylammonium chloride, cetylbenzyldimethylammonium chloride, cetylpyridinium bromide, cetyltrimethylammonium bromide, cetyldimethylethylammonium bromide, cetyltributylphosphonium bromide, dodecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide, and the like.

[0374] In some embodiments, the emulsion includes a quaternary ammonium compound, such as N-alkyldimethylbenzylammonium saccharinate, 1,3,5-triazine-1,3,5(2H,4H,6H)-triethanol, 1-decaneaminium, N-decyl-N,N-dimethyl-, chloride (or) didecyldimethylammonium chloride, 2-(2-(p-(diisobuyl)cresosoxy)ethoxy)ethyldimethylbenzylammonium chloride, 2-(2-(p-(diisobutyl)phenoxy)ethoxy)ethyldimethylammonium chloride, Alkyl benzyl ammonium chloride;Alkyl 1 or 3 benzyl-1-(2-hydroxyethyl)-2-imidazolinium chloride;Alkyl bis(2-hydroxyethyl)benzyl ammonium chloride;Alkyl dimethyl benzyl ammonium chloride;Alkyl dimethyl 3,4-dichlorobenzyl ammonium chloride (100% C12);Alkyl dimethyl 3,4-dichlorobenzyl ammonium chloride (50% C14, 40% C12, 10% C16);Alkyl dimethyl 3,4-dichlorobenzyl ammonium chloride Chlorobenzylammonium chloride (55% C14, 23% C12, 20% C16); alkyldimethylbenzylammonium chloride; alkyldimethylbenzylammonium chloride (100% C14); alkyldimethylbenzylammonium chloride (100% C16); alkyldimethylbenzylammonium chloride (41% C14, 28% C12); alkyldimethylbenzylammonium chloride (47% C12, 18% C14); alkyldimethylbenzylammonium chloride (55% C16, 2 0% C14); alkyldimethylbenzylammonium chloride (58% C14, 28% C16); alkyldimethylbenzylammonium chloride (60% C14, 25% C12); alkyldimethylbenzylammonium chloride (61% C11, 23% C14); alkyldimethylbenzylammonium chloride (61% C12, 23% C14); alkyldimethylbenzylammonium chloride (65% C12, 25% C14); alkyldimethylbenzylammonium chloride (67% C12, 24% C14);Alkyldimethylbenzylammonium chloride (67% C12, 25% C14); Alkyldimethylbenzylammonium chloride (90% C14, 5% C12); Alkyldimethylbenzylammonium chloride (93% C14, 4% C12); Alkyldimethylbenzylammonium chloride (95% C16, 5% C18); Alkyldimethylbenzylammonium chloride (and) Didecyldimethylammonium chloride; Alkyldimethylbenzylammonium chloride (such as in fatty acids); Alkyldimethylbenzylammonium Alkyldimethylbenzylammonium chlorides (C12-C16); Alkyldimethylbenzyl and dialkyldimethylammonium chlorides; Alkyldimethyldimethyldimethybenzylammonium chloride; Alkyldimethylethylammonium bromides (90% C14, 5% C16, 5% C12); Alkyldimethylethylammonium bromides (mixed alkyl and alkenyl groups, such as those in the fatty acids of soybean oil); Alkyldimethylethylbenzylammonium chloride Alkyldimethylethylbenzylammonium chloride (60% C14); Alkyldimethylisoproylbenzyl(isoproylbenzyl)ammonium chloride (50% C12, 30% C14, 17% C16, 3% C18); Alkyltrimethylammonium chloride (58% C18, 40% C16, 1% C14, 1% C12); Alkyltrimethylammonium chloride (90% C18, 10% C16); Alkyldimethyl(ethylbenzyl)ammonium chloride (C12-18); Di-(C8-10)-Alkyldimethyl ammonium chloride;Dialkyldimethylammonium chloride;Dialkyldimethylammonium chloride;Dialkyldimethylammonium chloride;Dialkylmethylbenzylammonium chloride;Didecyldimethylammonium chloride;Diisodecyldimethylammonium chloride;Dioctyldimethylammonium chloride;Dodecylbis(2-hydroxyethyl)octylhydrogenammonium chloride;Dodecyldimethylbenzylammonium chloride;Dodecylcarbamoylmethyldimethylbenzylammonium chlorideHeptadecylhydroxyethylimidazolinium chloride;Hexahydro-1,3,5-thris(2-hydroxyethyl)-s-triazine;Myristalkonium chloride (and) Quaternium-14;N,N-Dimethyl-2-hydroxypropylammonium chloride polymer;n-Alkyldimethylbenzylammonium chloride;n-Alkyldimethylethylbenzylammonium chloride;n-Tetradecyldimethylbenzylammonium chloride monohydrate;Octyldecyldimethylammonium chloride;Octyldodecyldimethylammonium chloride;Octyphenoxyethoxyethyl(octyphenoxyethoxyethyl)dimethylbenzylammonium chloride;Oxydiethylenebis(alkyldimethylammonium chloride);Quaternary ammonium compounds, dicocoalkyldimethyl, chloride;Trimethoxysilypropyldimethyloctadecylammonium chloride;Trimethoxysilylquat quats), trimethyldodecylbenzyl ammonium chloride; n-dodecyldimethylethylbenzyl ammonium chloride; n-hexadecyldimethylbenzyl ammonium chloride; n-tetradecyldimethylbenzyl ammonium chloride; n-tetradecyldimethylethylbenzyl ammonium chloride; and n-octadecyldimethylbenzyl ammonium chloride.

[0375] Nanoemulsion formulations and methods of making such formulations are well known to those skilled in the art and are described, for example, in U.S. Pat. Nos. 7,476,393, 7,468,402, 7,314,624, 6,998,426, 6,902,737, 6,689,371, 6,541,018, 6,464,990, 6,461,625, and 6,419,946. , 6,413,527, 6,375,960, 6,335,022, 6,274,150, 6,120,778, 6,039,936, 5,925,341, 5,753,241, 5,698,219, 5,152,923 and Fanun et al. (2009), Microemulsions: Properties and Applications (Surfactant Science), CRC Press, Boca Raton FL.

[0376] In some embodiments, one or more active agents described herein may be provided as a "concentrate," for example, in a storage container (e.g., in a pre-measured volume) ready for dilution or in a dissolvable capsule ready to be added to a volume of water, alcohol, hydrogen peroxide, or other diluent.

[0377] In some embodiments, the compounds of Formula (I) or Formula (III) described herein are formulated as inclusion complexes. While not limited to cyclodextrin inclusion complexes, it is noted that cyclodextrins are the most frequently used agents for forming inclusion complexes of pharmaceuticals. Cyclodextrins (CDs) are cyclic oligomers of glucose, typically containing 6, 7, or 8 glucose monomers joined by α-1,4 linkages. These oligomers are commonly referred to as α-CD, β-CD, and γ-CD, respectively. Higher oligomers containing up to 12 glucose monomers are known and are contemplated in the formulations described herein. Functionalized cyclodextrin inclusion complexes are also contemplated. Exemplary, but non-limiting, functionalized cyclodextrins include sulfonates, sulfonates and sulfinates, or disulfonates of hydroxybutenyl cyclodextrin; sulfonates, sulfonates and sulfinates, or disulfonates of mixed ethers of cyclodextrin, in which at least one of the ether substituents is hydroxybutenyl cyclodextrin. Exemplary cyclodextrins include polysaccharide ethers containing at least one 2-hydroxybutenyl substituent, in which at least one hydroxybutenyl substituent is sulfonated and sulfinated or disulfonated, and alkyl polyglycoside ethers containing at least one 2-hydroxybutenyl substituent, in which at least one hydroxybutenyl substituent is sulfonated and sulfinated or disulfonated. In various embodiments, inclusion complexes formed between sulfonated hydroxybutenyl cyclodextrin and one or more active agents described herein are contemplated. Methods for preparing cyclodextrins and cyclodextrin inclusion complexes can be found, for example, in U.S. Patent Publication No. 2004 / 0054164 and references cited therein, and in U.S. Patent Publication No. 2011 / 0218173 and references cited therein.

[0378] In certain embodiments, the compounds of Formula (I) or Formula (III) described herein can be administered using medical devices known in the art. For example, in one embodiment, the pharmaceutical compositions of the present invention can be administered using needleless hypodermic injection devices, such as those disclosed in U.S. Patent No. 5,399,163, U.S. Patent No. 5,383,851, U.S. Patent No. 5,312,335, U.S. Patent No. 5,064,413, U.S. Patent No. 4,941,880, U.S. Patent No. 4,790,824, or U.S. Patent No. 4,596,556. Examples of well-known implants and modules useful for such delivery include, but are not limited to, U.S. Patent No. 4,487,603, which discloses an implantable microinjection pump for dispensing drugs at a controlled rate; U.S. Patent No. 4,486,194, which discloses a therapeutic device for administering pharmaceuticals through the skin; U.S. Patent No. 4,447,233, which discloses a drug injection pump for delivering drugs at a precise injection rate; U.S. Patent No. 4,447,224, which discloses a variable flow implantable infusion device for continuous drug delivery; U.S. Patent No. 4,439,196, which discloses an osmotic drug delivery system with multi-chamber compartments; and U.S. Patent No. 4,475,196, which discloses an osmotic drug delivery system. In certain embodiments, the compound of formula (I) or formula (III) can be administered using a drug-eluting stent, such as those described in WO01 / 87263 and related publications or those corresponding to those described by Perin (Perin, EC, 2005). Many other such implants, delivery systems and modules are known to those skilled in the art.

[0379] The dosage of the compounds of formula (I) or formula (III) described herein will vary depending on the particular compound, the disease involved, the subject, and the nature and severity of the disease, as well as the physical condition of the subject, and the selected route of administration. The appropriate dosage can be easily determined by one skilled in the art.

[0380] In certain embodiments, the composition may contain from 0.1% to, for example, 0.1-70%, or 5-60%, or 10-30%, of one or more compounds of formula (I) or formula (III), depending on the method of administration.

[0381] Those skilled in the art will understand that the optimal amount and interval of each administration of the compounds of formula (I) or formula (III) described herein will be determined by the nature and magnitude of the condition to be treated, the form, route and site of administration, and the age and condition of the particular subject to be treated, and that the appropriate dosage to be used will be ultimately determined by a physician. This dosage may be repeated as appropriate. If side effects occur, the amount and / or frequency of administration may be changed or reduced in accordance with normal clinical practice.

[0382] In some embodiments, pharmaceutically acceptable salts can include both acid and base addition salts. In some embodiments, a "pharmaceutically acceptable acid addition salt" retains the biological effectiveness and properties of the free base, is not biologically or otherwise undesirable, and is selected from inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, as well as organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, and galactaric acid. , gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like.

[0383] In some embodiments, "pharmaceutically acceptable base addition salts" can refer to salts that retain the biological effectiveness and properties of the free acid and are not biologically or otherwise undesirable. These salts can be prepared from the addition of an inorganic or organic base to the free acid. Salts derived from inorganic bases can include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. For example, inorganic salts can include, but are not limited to, ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases may include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like.

[0384] For purposes of administration, the compounds of the present disclosure can be administered as raw chemicals or formulated as pharmaceutical compositions. Pharmaceutical compositions of the present disclosure include one or more compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, and a pharmaceutically acceptable carrier. One or more compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, are present in the composition in an amount effective to treat the specific disease or condition in question, for example, in an amount sufficient to treat a bacterial infection, and with a toxicity generally acceptable to the subject. In some embodiments, one or more compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, are present in the composition in an amount effective to treat the specific disease or condition in question. The antibacterial activity of a compound of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer of either thereof, can be determined by one skilled in the art, for example, as described in the Examples herein. Appropriate concentrations and dosages can be readily determined by one skilled in the art.

[0385] Administration of one or more compounds of the present disclosure, such as a compound of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer of any of these, in pure form or in a suitable pharmaceutical composition, can be carried out via any of the accepted modes of administration of agents to fulfill similar utilities. In some embodiments, the one or more compounds of the present disclosure administered are a compound of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer of any of the foregoing, in pure form or in a suitable pharmaceutical composition.

[0386] Pharmaceutical compositions of the present disclosure can be prepared by combining one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof with a suitable pharmaceutically acceptable carrier, and formulated into preparations in solid, semi-solid, liquid, or gaseous forms, such as tablets, capsules, powders, granules, ointments, liquids, suppositories, injections, inhalants, gels, microspheres, and aerosols. Typical routes of administration of such pharmaceutical compositions include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal. As used herein, the term parenteral can include subcutaneous injection, intravenous, intramuscular, intrasternal injection, or injection techniques. Pharmaceutical compositions of the present disclosure can be formulated to make the active ingredients contained therein bioavailable when the composition is administered to a subject. The composition administered to a subject or patient may take the form of one or more dosage units; for example, a tablet may be a single dosage unit, and a container of one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, in aerosol form may hold multiple dosage units. Actual methods for preparing such dosage forms will be known or apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000).

[0387] The pharmaceutical compositions of the present disclosure may be in solid or liquid form. In one embodiment, the carrier is particulate, so that the composition is, for example, in tablet or powder form. The carrier may be liquid, so that the composition is, for example, an oral syrup, an injectable liquid, or an aerosol, which may be useful, for example, for inhalatory administration.

[0388] When intended for oral administration, the pharmaceutical compositions of the present disclosure will typically be either solid or liquid, with semi-solid, semi-liquid, suspension, and gel forms being included among the forms considered herein as either solid or liquid.

[0389] As a solid composition for oral administration, the pharmaceutical composition may be formulated into the form of powder, granules, compressed tablets, pills, capsules, chewing gum, wafers, etc. Such solid compositions typically contain one or more inert diluents or edible carriers. Additionally, one or more of the following may be present: binders such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, tragacanth gum, or gelatin; excipients such as starch, lactose, or dextrin; disintegrants such as alginic acid, sodium alginate, Primojel®, corn starch, etc.; lubricants such as magnesium stearate or Sterotex®; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; flavorings such as peppermint, methyl salicylate, or orange flavoring; and coloring agents.

[0390] When the pharmaceutical composition is in the form of a capsule, for example, a gelatin capsule, it may contain, in addition to the materials disclosed herein, a liquid carrier such as polyethylene glycol or oil.

[0391] The pharmaceutical compositions of the present disclosure may be in the form of a liquid, such as an elixir, syrup, solution, emulsion, or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, the pharmaceutical compositions of the present disclosure typically contain one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, as well as one or more sweeteners, preservatives, dyes / colorants, and flavor enhancers. In compositions intended for injection administration, one or more of surfactants, preservatives, wetting agents, dispersing agents, suspending agents, buffers, stabilizers, and isotonic agents may be included.

[0392] Liquid pharmaceutical compositions of the present disclosure, whether in solution, suspension, or other similar form, may contain one or more of the following adjuvants: sterile diluents, such as water for injection, saline solution, physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono- or diglycerides, polyethylene glycol, glycerin, propylene glycol, or other solvents that may serve as solvents or suspension media; antibacterial agents, such as benzyl alcohol or methylparabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, or phosphates, as well as agents for adjusting isotonicity, such as sodium chloride or dextrose. Parenteral preparations may be enclosed in ampoules, disposable syringes, or multiple-dose vials made of glass or plastic. In some embodiments, the adjuvant is saline. In some embodiments, the injectable pharmaceutical composition is sterile.

[0393] Liquid pharmaceutical compositions of the present disclosure intended for either parenteral or oral administration should contain an amount of one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof so that a suitable dosage will be obtained.

[0394] The pharmaceutical compositions of the present disclosure may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment, or gel base. The base may comprise, for example, one or more of the following: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickeners may be present in pharmaceutical compositions for topical administration. If intended for transdermal administration, the composition may comprise a transdermal patch or iontophoresis device.

[0395] The pharmaceutical composition of the present disclosure may be intended for rectal administration, for example, in the form of a suppository that melts in the rectum and releases the drug. Compositions for rectal administration may contain an oily base as a suitable non-irritating excipient. Such bases may include, but are not limited to, lanolin, cocoa butter, and polyethylene glycol.

[0396] The pharmaceutical compositions of the present disclosure may contain various materials that modify the physical form of a solid or liquid dosage unit. For example, the compositions may contain materials that form a coating shell around the active ingredient. The materials that form the coating shell are typically inert and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be encapsulated in a gelatin capsule.

[0397] Pharmaceutical compositions of the present disclosure may include an agent that binds to one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, in solid or liquid form, thereby aiding in the delivery of the one or more compounds, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof. Suitable agents that may act in this capacity may include monoclonal or polyclonal antibodies, proteins, or liposomes.

[0398] The pharmaceutical compositions of the present disclosure may be prepared in dosage units that can be administered as an aerosol. The term aerosol is used to describe a variety of systems, ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by liquefied or compressed gas or by a suitable pump system that dispenses the active ingredient. Aerosols of one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof may be delivered in single-phase, two-phase, or three-phase systems to deliver the active ingredient. Aerosol delivery may include the necessary container, activator, valve, subcontainer, etc., which may together form a kit. One of ordinary skill in the art can determine a preferred aerosol without undue experimentation.

[0399] Pharmaceutical compositions of the present disclosure can be prepared by methodologies well known in the pharmaceutical arts. For example, pharmaceutical compositions intended to be administered by injection can be prepared by combining one or more compounds of the present disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers with sterile distilled water to form a solution. A surfactant can be added to facilitate the formation of a homogeneous solution or suspension. A surfactant is a compound that non-covalently interacts with one or more compounds of the present disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers to facilitate the dissolution and homogeneous suspension of one or more compounds of the present disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers in an aqueous delivery system.

[0400] In some embodiments, pharmaceutical compositions of the present disclosure can be enriched primarily to obtain one enantiomer of a compound described herein. An enantiomer-enriched mixture can contain, for example, at least 60 mol percent, or at least 75, at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, at least 99.5, or even 100 mol percent of one enantiomer. In some embodiments, compositions described herein enriched with one enantiomer can be substantially free of the other enantiomer, where substantially free can mean, for example, that the substance constitutes less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% of the amount of the other enantiomer in the pharmaceutical composition or mixture of compounds. For example, if a pharmaceutical composition or mixture of compounds contains 98 grams of a first enantiomer and 2 grams of a second enantiomer, it is said to contain 98 mol percent of the first enantiomer and only 2 mol percent of the second enantiomer.

[0401] In some embodiments, pharmaceutical compositions of the present disclosure can be enriched primarily to obtain a diastereoisomer of a compound disclosed herein. A diastereoisomer-enriched mixture can contain, for example, at least 60 mol percent, or at least 75, at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, at least 99.5, or even 100 mol percent of a diastereoisomer. In some embodiments, a diastereoisomer-enriched composition described herein can be substantially free of other diastereoisomers, where substantially free can mean, for example, that the substance constitutes less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% of the amount of other diastereoisomers in the pharmaceutical composition or mixture of compounds.

[0402] Diastereomeric mixtures can be separated into individual diastereoisomers based on their physical chemical differences by methods well known to those skilled in the art, such as, for example, chromatography and / or fractional crystallization. Enantiomers can also be separated by converting the enantiomeric mixture to a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereoisomers, and converting the individual diastereoisomers to their corresponding pure enantiomers (e.g., by hydrolysis). Enantiomers can also be separated using chiral HPLC columns. Some of the compounds of the present disclosure may also be in atropisomeric or rotamer forms and are considered part of the present disclosure.

[0403] therapeutic use The compounds, compositions and methods of the present disclosure may be useful in several different subjects, including, but not limited to, mammals, humans, non-human mammals, domesticated animals (e.g., laboratory animals, pets or farm animals), non-domesticated animals (e.g., wildlife), dogs, cats, rodents, mice, hamsters, cows, birds, chickens, fish, pigs, horses, goats, sheep, rabbits, or any combination thereof.

[0404] The compounds and compositions described herein can be useful as therapeutic agents, e.g., for treatment, where they can be administered to a subject in need thereof. The therapeutic benefit of the present disclosure can be achieved by the reduction, suppression, amelioration, or eradication of a disease state in a subject, including, but not limited to, the symptoms of the disease state. The therapeutic benefit in a subject having a disease or condition, or predisposed to or beginning to have a disease or condition, can be achieved by the reduction, suppression, prevention, amelioration, or eradication of the condition or disease, or a pre-condition or pre-disease state.

[0405] In practicing the methods described herein, a therapeutically effective amount of a composition described herein can be administered to a subject in need thereof, often to treat and / or prevent the condition or progression thereof. The pharmaceutical composition can affect the subject's physiological function, such as the immune system, inflammatory response, or other physiological effects. The therapeutically effective amount can vary depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors.

[0406] Treat and / or treating can refer to any evidence of successful treatment or alleviation of a disease or condition. Treating can include, for example, reducing the severity, delaying, or alleviating one or more symptoms of a disease or condition, or reducing the frequency with which a patient experiences symptoms of a disease, deficiency, disorder, or adverse condition. Treating can be used herein to refer to a method that results in some level of treatment or alleviation of a disease or condition and can envision a variety of outcomes toward that goal, including, but not limited to, preventing the condition altogether.

[0407] Prevent, preventing, etc. can refer to preventing a disease or condition, such as tumor formation, in a patient. For example, if an individual at risk of developing a tumor or other form of cancer is treated with the methods of the present disclosure and does not subsequently develop a tumor or other form of cancer, then the disease has been prevented in that individual for at least some period of time. Preventing can also refer to preventing the recurrence of a disease or condition in a patient who has previously been treated for the disease or condition, for example, by preventing recurrence.

[0408] A therapeutically effective amount can be an amount of a composition or its active ingredients sufficient to achieve a beneficial effect on an individual to whom the composition is administered, or otherwise reduce undesirable or unfavorable events. A therapeutically effective dose can be a dose that produces one or more desired or desirable (e.g., beneficial) effects, administered one or more times over a given period of time. The exact dose can be determined by the purpose of treatment and can be ascertained by one of ordinary skill in the art using known techniques.

[0409] Methods for Treating Bacterial Infections The present disclosure features methods of treating bacterial infections, and uses for treating bacterial infections, comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, or a therapeutically effective amount of a pharmaceutical composition comprising one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof.

[0410] The compounds or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof and pharmaceutical compositions of the present disclosure (e.g., compounds of Formula (I) or Formula (III)) may have antibacterial activity against a wide range of Gram-positive and Gram-negative bacteria. Representative susceptible organisms may include Gram-positive, Gram-negative, aerobic, and facultative anaerobic organisms whose growth can be inhibited by the compounds of the present disclosure, including, but not limited to, Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Enterococcus, Burkholderia, and other organisms.

[0411] Thus, the compounds or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and pharmaceutical compositions comprising the compounds disclosed herein or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, can be used, for example, to treat bacterial infections and conditions caused by bacterial infections. Bacterial infections amenable to treatment according to the present disclosure can include primary infections and co-infections caused by certain bacteria, as well as one or more additional infectious agents, such as bacteria, viruses, parasites, and fungi.

[0412] In one aspect, there is provided a method of treating a bacterial infection, comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer of any thereof, or a pharmaceutical composition comprising one or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer of any thereof.

[0413] In some embodiments, the bacterial infection is caused by Staphylococcus sp., Lactobacillus sp., Streptococcus sp., Sarcina sp., Escherichia sp., Enterobacter sp., Klebsiella sp., Pseudomonas sp., Acinetobacter sp., Mycobacterium sp., Proteus sp., Campylobacter sp., Citrobacter sp., Neisseria sp., Bacillus sp. Infections caused by Peptococcus sp., Salmonella sp., Shigella sp., Serratia sp., Haemophilus sp., Brucella sp., Francisella sp., Yersinia sp., Corynebacterium sp., Moraxella sp., Burkholderia sp., or Enterococcus sp.

[0414] In some embodiments, the bacterial infection is due to a gram-positive, gram-negative, aerobic, or facultative anaerobic bacteria.

[0415] In some embodiments, the bacterial infection is caused by a Gram-positive bacterium. Examples of Gram-positive bacteria include, but are not limited to, Staphylococcus species, Lactobacillus species, Streptococcus species, Sarcina species, Bacillus species, Peptococcus species, Corynebacterium species, and Enterococcus species. In some embodiments, the Gram-positive bacterial infection is an infection by Staphylococcus species, Lactobacillus species, Streptococcus species, Sarcina species, Bacillus species, Peptococcus species, Corynebacterium species, or Enterococcus species. In some embodiments, the bacterial infection is caused by Staphylococcus aureus. In some embodiments, the bacterial infection is caused by methicillin-resistant Staphylococcus aureus (MRSA).

[0416] In some embodiments, the bacterial infection is caused by gram-negative bacteria. Examples of gram-negative bacteria include, but are not limited to, Escherichia spp., Enterobacter spp., Klebsiella spp., Pseudomonas spp., Acinetobacter spp., Proteus spp., Campylobacter spp., Citrobacter spp., Neisseria spp., Salmonella spp., Shigella spp., Serratia spp., Haemophilus spp., Brucella spp., Francisella spp., Yersinia spp., Burkholderia spp., and Moraxella spp.

[0417] In some embodiments, the bacterial infection is caused by a fermentative or non-fermentative gram-negative bacteria. Examples of fermentative or non-fermentative Gram-negative bacteria include, but are not limited to, Pseudomonas aeruginosa; Stenotrophomonas maltophila; Burkholderia cepacia; Alcaligenes xylosoxidans; Enterobacteriaceae; Haemophilus spp.; Franciscellaceae (e.g., Francisella tularensis); Neisseria spp.; and Enterobacteriaceae, such as Serratia spp., Proteus spp., Klebsiella spp., Enterobacter spp., Citrobacter spp., Salmonella spp., Providencia spp., Yersinia spp. (e.g., Yersinia pestis), Morganella spp., Cedecea spp., Edwardsiella spp., Acinetobacter spp., and Escherichia coli.

[0418] In some embodiments, the Gram-negative bacterial infection is an infection caused by Escherichia spp., Enterobacter spp., Klebsiella spp., Pseudomonas spp., Acinetobacter spp., Proteus spp., Campylobacter spp., Citrobacter spp., Neisseria spp., Salmonella spp., Shigella spp., Serratia spp., Haemophilus spp., Brucella spp., Francisella spp., Yersinia spp., Burkholderia spp., or Moraxella spp. In some embodiments, the Gram-negative bacterial infection is an infection caused by Enterobacteriaceae spp., Haemophilus spp., Francisellaceae spp., or Neisseria spp. In some embodiments, the bacterial infection is caused by Enterobacteriaceae spp. In some embodiments, the bacterial infection is caused by extended-spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae spp.

[0419] In some embodiments, the Gram-negative bacterial infection is an infection caused by Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, Proteus mirabilis, Bacillus anthracis, P. aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei, or Yersinia pestis. In some embodiments, the Gram-negative bacterial infection is an infection caused by Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxydans.

[0420] In some embodiments, the bacterial infection is caused by a multidrug-resistant (MDR) organism. In some embodiments, the bacterial infection is caused by a multidrug-resistant biothreat pathogen, including those defined as such by the U.S. Centers for Disease Control and Prevention's Category A / B / C bioterrorism agent designation. In some such embodiments, the biothreat pathogen is a Gram-negative bacterium.

[0421] In some embodiments, one or more compounds of Formula (I) or Formula (III) or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, or pharmaceutical compositions of the present disclosure are useful in treating skin infections; pneumonia; hospital-acquired pneumonia; sepsis; respiratory tract infections (e.g., lower or upper respiratory tract infections); pulmonary infections in cystic fibrosis patients; acute exacerbation of chronic bronchitis; community-acquired pneumonia; hospital-acquired pneumonia (including ventilator-associated pneumonia (VAP)); hospital-acquired or community-acquired infections caused by Gram-negative bacteria; upper respiratory tract infections Diseases; diffuse panbronchiolitis; tonsillitis; pharyngitis; otitis including acute sinusitis and mastoiditis; urinary tract and genital infections, e.g. complicated urinary tract infections, cystitis, resistant or recurrent cystitis, urethritis, pyelonephritis, acute pyelonephritis, endometritis, prostatitis, salpingitis and epididymitis; eye infections, e.g. conjunctivitis, corneal ulcers, iridocyclitis and postoperative infections in radial keratotomy patients; blood infections, e.g. sepsis or bacteremia; skin and soft tissue infections, e.g. infectious dermatitis bone and joint infections, such as osteomyelitis and septic arthritis; gastrointestinal infections, such as dysentery, enteritis, colitis, necrotizing enterocolitis, and anorectal infections; intra-abdominal infections, including, but not limited to, complicated intra-abdominal infections, typhoid fever, infectious diarrhea, peritonitis with appendicitis, pelvic peritonitis, and intra-abdominal abscesses; infections in the mouth, such as infections after dental surgery; other infections, such as melioidosis, infective endocarditis, sexually transmitted diseases, liver abscess, cholecystitis, cholangitis, mastitis, and meningitis caused by gram-negative or gram-positive bacteria, and infections of the nervous system, as disclosed herein.

[0422] In some embodiments, the methods disclosed herein treat an infection selected from the group consisting of complicated intra-abdominal infection, complicated urinary tract infection, resistant or recurrent cystitis, acute pyelonephritis, and bacteremia.

[0423] In a further aspect, the present disclosure provides the use of one or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof to treat a bacterial infection as described herein in a subject in need thereof.

[0424] In yet a further aspect, provided herein is one or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof for use in a method of treating a bacterial infection in a subject in need thereof.

[0425] In another aspect, the disclosure provides one or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer of any of the foregoing for use as a medicament.

[0426] In yet a further aspect, provided herein is the use of one or more compounds of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a subject in need of treatment for a bacterial infection.

[0427] In still a further aspect, provided herein is one or more pharmaceutical compositions of the present disclosure for use in a method of treating a bacterial infection in a subject in need thereof.

[0428] In yet a further aspect, provided herein is the use of one or more pharmaceutical compositions of the present disclosure in the manufacture of a medicament for treating a subject in need of treatment for a bacterial infection.

[0429] In any of the foregoing methods and uses disclosed herein, the subject may be a mammal. In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human.

[0430] Administration One or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer of either thereof, or a composition of any thereof, may be administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the particular compound employed; the metabolic stability and length of action of the compound; the age, weight, general health, sex, and diet of the subject; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disorder or condition; and the subject being treated.

[0431] Administration of one or more compounds of the present disclosure, such as a compound of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer of any of these, in pure form or in a suitable pharmaceutical composition, can be carried out via any of the accepted modes of administration of agents to fulfill similar utilities. In some embodiments, the one or more compounds of the present disclosure administered are a compound of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer of any of the foregoing, in pure form or in a suitable pharmaceutical composition.

[0432] The pharmaceutical compositions may be used in the methods described herein and may be administered to a subject in need thereof using techniques known to those of skill in the art that may be suitable as a treatment for a disease or condition affecting the subject. Those of skill in the art will understand that the amount, duration, and frequency of administration of the pharmaceutical compositions described herein to a subject in need thereof will depend on several factors, including, but not limited to, the health of the subject, the patient's particular disease or condition, the grade or level of the patient's particular disease or condition, additional therapeutic agents the subject is receiving or has received, etc.

[0433] The methods and compositions described herein can be for administration to a subject in need thereof. In many cases, administration of the compositions described herein can include a route of administration, non-limiting examples of which include oral, intravenous, intra-arterial, subcutaneous, subdural, intramuscular, intracranial, intrasternal, intratumoral, or intraperitoneal. Additionally, pharmaceutical compositions can be administered to a subject by additional routes of administration, for example, by inhalation, dermal, intranasal, or intrathecal administration.

[0434] The compositions of the present disclosure may be administered to a subject in need thereof in a first dose and one or more additional doses. The one or more additional doses may be administered to a subject in need thereof minutes, hours, days, weeks, or months after the first dose. Any one of the additional doses may be administered to a subject in need thereof less than 21 days, less than 14 days, less than 10 days, less than 7 days, less than 4 days, or less than 1 day after the first dose. The one or more doses may occur more than once per day, more than once per week, or more than once per month. Administration may be weekly, biweekly (every two weeks), every three weeks, monthly, or bimonthly.

[0435] One or more compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, may be administered simultaneously with, before, or after the administration of one or more other therapeutic agents. Such combination therapy may include the administration of a single pharmaceutical formulation containing one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and one or more additional active agents, as well as the administration of one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and each active agent in its own separate pharmaceutical formulation. For example, one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and the other active agent(s) may be administered to a subject together in a single oral administration composition, such as a tablet or capsule, or each agent may be administered in a separate oral formulation. When separate formulations are used, one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof and one or more additional active agents may be administered at essentially the same time, e.g., simultaneously, or at different staggered times, e.g., sequentially, and combination therapy is understood to include all of these regimens.

[0436] kit In some aspects, the present disclosure provides kits comprising a compound disclosed herein or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof, or a composition comprising said compound or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof and instructions.

[0437] In some embodiments, the present disclosure provides a kit comprising a pharmaceutical composition disclosed herein and instructions.

[0438] In some embodiments, the instructions include instructions for administering the disclosed compound, salt, or pharmaceutical composition to a subject in need thereof. In some embodiments, the kit includes a compound, salt, or pharmaceutical composition disclosed herein packaged with a desiccant in a low moisture vapor permeable container. Optionally, a label is on or associated with the container. For example, a label is on a container when letters, numbers, or other symbols forming the label are affixed, molded, or engraved onto the container itself, and is associated with a container when the label is present, e.g., as a package insert, in a receptacle or carrier, e.g., a box that also holds the container. Furthermore, a label can be used to indicate that the contents are to be used for a specific therapeutic application. In some embodiments, the label includes instructions for using the contents, e.g., in a method described herein. In some embodiments, the compound, salt, or pharmaceutical composition disclosed herein is presented in a pack or container containing one or more unit dosage forms comprising the compound, salt, or pharmaceutical composition disclosed herein. The pack can contain metal or plastic foil, e.g., a blister pack. The pack or container can be accompanied by instructions for administering the unit dosage form. In some embodiments, the pack or container is accompanied by notices in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which notice reflects the agency's approval of the drug form for human or veterinary administration. Such notices may be, for example, labeling or an approved product insert approved by the U.S. Food and Drug Administration to regulate the drug. In some embodiments, compositions comprising the compounds, salts, or pharmaceutical compositions disclosed herein are prepared, placed in an appropriate container, and labeled for treating an indicated condition.

[0439] As used herein, a "kit" can include a container for containing at least one pharmaceutical composition or compound of the present disclosure, and can also include divided containers, such as divided bottles or divided foil packets. The container can be of any conventional shape or form known in the art, made of pharmaceutically acceptable materials, such as a paper or cardboard box, a glass or plastic bottle or jar, a resealable bag (e.g., holding tablet "refills" to be placed in different containers), or a blister pack with individual doses that are pushed out of the pack according to the treatment schedule. The container used can depend on the exact dosage form contained; for example, a traditional cardboard box is not generally used to hold liquid suspensions. It is feasible to use more than one container together in a single package to sell a single dosage form. For example, tablets can be contained in a bottle, which in turn can be contained in a box.

[0440] An example of such a kit is a so-called blister pack. Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms (tablets, capsules, etc.). Blister packs generally consist of a sheet of relatively stiff material covered with a foil of transparent plastic material. During the packaging process, depressions are formed in the plastic foil. The depressions may have the size and shape of individual tablets or capsules to be packed, or may have a size and shape adapted for packing multiple tablets and / or capsules. The tablets or capsules are then appropriately placed into the depressions, and a sheet of relatively stiff material is sealed against the plastic foil on the side of the plastic foil opposite the direction in which the depressions were formed. As a result, the tablets or capsules are individually sealed in the depressions between the plastic foil and the sheet, or collectively sealed if desired. In some embodiments, the strength of the sheet is such that the tablets or capsules can be removed from the blister pack by manually applying pressure to the depressions, thereby forming openings in the sheet at the locations of the depressions. The tablet or capsule can then be removed through the opening.

[0441] Another specific embodiment of the kit is a dispenser designed to dispense the daily doses one at a time in the order of their intended use. In some embodiments, the dispenser is equipped with a memory aid to further facilitate compliance with the regimen. An example of such a memory aid is a mechanical counter that indicates the number of daily doses dispensed. Another example of such a memory aid is a battery-powered microchip memory in conjunction with a liquid crystal display or an audio reminder signal, for example, that reads out the date the last daily dose was taken and / or reminds the user of the date the next dose should be taken.

[0442] The kits of the present disclosure may also include one or more additional pharmaceutically active compounds or pharmaceutical compositions in addition to one or more compounds or pharmaceutical compositions of the present disclosure. For example, the additional compound may be a second antibacterial compound, or the additional pharmaceutical composition may include a second antibacterial compound. The additional compound or pharmaceutical composition may be administered in the same dosage form as the one or more compounds or pharmaceutical compositions of the present disclosure, or in a different dosage form. Similarly, the additional compound or pharmaceutical composition may be administered at the same time as the one or more compounds or pharmaceutical compositions of the present disclosure, or at a different time.

[0443] In some embodiments, kits of the present disclosure have notices in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals or biological products, which notice reflects (a) agency approval for manufacture, use, or sale for human administration, (b) instructions for use, or both.

[0444] Although the technology herein has been described with reference to particular embodiments, it should be understood that these embodiments are merely illustrative of the principles and applications of the technology. It will be apparent to those skilled in the art that various modifications and variations can be made to the methods and apparatus of the technology without departing from the spirit and scope of the technology. Accordingly, it is intended that the technology include modifications and variations that come within the scope of the appended claims and their equivalents. Accordingly, the present disclosure is not limited except as by the appended claims.

[0445] References throughout this specification to "one embodiment" or "an embodiment" mean that a particular feature, structure, or characteristic described in connection with an embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the phrase "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. All ranges cited herein are inclusive.

[0446] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous modifications, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention, and that methods and structures within the scope of these claims and their equivalents be covered thereby.

[0447] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0448] Aspects of the present technology are illustrated in more detail with reference to the following examples. Before describing some exemplary embodiments of the technology, it should be understood that the technology is not limited to the details of the configuration or process steps set forth in the following description. The technology enables other embodiments and can be practiced or carried out in various ways. The following examples are set forth to illustrate certain aspects of the present technology and are not intended to limit them. It is understood that one of ordinary skill in the art could potentially make these compounds by similar methods or by combining other methods known to those of ordinary skill in the art. It is also understood that one of ordinary skill in the art could make additional compounds within the scope of the present disclosure by using appropriate starting materials and modifying the synthetic routes as necessary in a similar manner to those described below. In general, starting materials and reagents can be obtained from commercial vendors or synthesized according to sources known to those of ordinary skill in the art or prepared as described herein.

[0449] Reagent / reactant names shown are as labeled on the commercial bottle or as generated by IUPAC conventions, ChemDraw 19.0 (CAMBRIDGESOFT®; PerkinElmer). Compounds designated as salts (e.g., hydrochloride, acetate, sulfate) may contain more than one molar equivalent of acid. [Example]

[0450] Abbreviation The following abbreviations are used herein: Ac Acetate or Acetyl Ac2O acetic anhydride AcOH glacial acetic acid AIBN Azobisisobutyronitrile BF3·OEt2 Boron trifluoride etherate Bn Benzyl BnBr benzyl bromide Boc t-butoxycarbonyl BSA Bovine serum albumin Bu3SnH Tri-n-butyltin hydride Cbz Carboxybenzyl CCl3CN Trichloroacetonitrile CsF Cesium Fluoride d double line DAST Diaminosulfur trifluoride DCM dichloromethane (methylene chloride) dd double line double line DIAD Diisopropyl azodicarboxylate DMAP N,N-dimethylaminopyridine DMEM Dulbecco's Modified Eagle's Medium DMF N,N-dimethylformamide DMSO dimethyl sulfoxide DMTMM Chloride 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride DPPA Diphenylphosphoryl azide equiv equivalent EtOAc ethyl acetate Et2O diethyl ether EtOH ethanol g grams h time HPLC High Pressure Liquid Chromatography LAH Lithium aluminum hydride LC / MS Liquid Chromatography / Mass Spectrometry LiOH Lithium hydroxide m multiplet M mole MeCN acetonitrile MeOH methyl alcohol min mmol millimole MPLC Medium Pressure Liquid Chromatography MS mass spectrometry n-normal (unbranched) N Normal NaH sodium hydride NaN3 Sodium Azide NaOMe Sodium methoxide NBS N-Bromosuccinimide NH4OAc Ammonium Acetate NMR nuclear magnetic resonance Pd(OH)2 palladium hydroxide pH -log[H + ] PhI(OAc)2 Phenyl iodonium acetate Piv Pivaloyl(trimethylacetyl) PMB para-methoxybenzyl PMB-Cl para-methoxybenzyl chloride ppm (parts per million) psi pounds per square inch PTSA para-toluenesulfonic acid RP-HPLC Reversed-phase high-pressure liquid chromatography R t retention time rt room temperature s Single line t triple line t- tertiary t-BuOH t-butanol TBAI Tetra-n-butylammonium iodide TBSCl t-Butyldimethylsilyl chloride TEA Triethylamine TfN3 Trifluoromethanesulfonyl azide TFA trifluoroacetic acid THF tetrahydrofuran TMS trimethylsilyl TMSOTf Trimethylsilyl trifluoromethanesulfonate TolSH p-Thiocresol TsCl para-toluenesulfonyl chloride USP United States Pharmacopoeia wt% weight percent

[0451] Analysis method Analytical data is included within the procedures below. Unless otherwise stated, all 1 H NMR data were collected on a Bruker Ultra Shield 500 MHz / 54 mm instrument (BZH43 / 500 / 70B, D221 / 54-3209) and chemical shifts are expressed in parts per million (ppm).

[0452] Example 1. Synthesis of (benzyl (((2S,5R)-5-azido-6-hydroxytetrahydro-2H-pyran-2-yl)methyl) (benzyl)carbamate (Ring A1).

[0453] [ka]

[0454] Step A. (S)-N-((E)-((S)-3,4-Dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinamide

[0455] [ka]

[0456] (S)-(+)-2-Methyl-2-propanesulfinamide (50 g, 412 mmol) and CuSO (131.7 g, 825 mmol) were mixed in DCM (600 mL). A solution of 2-formyl-3,4-dihydro-2H-pyran (48.6 g, 433 mmol, Synthonix) in DCM (100 mL) was added over 20 min. The mixture was stirred at room temperature for 18 h and then filtered through diatomaceous earth and rinsed with DCM. The filtrate was evaporated under reduced pressure. The material was purified on silica gel (5 × 330 g, dry loading) by MPLC using 0–20% EtO in hexanes as the eluent to give the title compound (23.74 g, 27%, second-eluting diastereoisomer). 1 H NMR (400 MHz, CDCl3) δ 8.08 (d, J = 3.0 Hz, 1H), 6.47 (dt, J = 6.3, 1.6 Hz, 1H), 4.81 - 4.72 (m, 2H), 2.19 - 1.94 (m, 4H), 1.24 (s, 9H).

[0457] Step B. (R)-N-(((S)-3,4-Dihydro-2H-pyran-2-yl)methyl)-2-methylpropane-2-sulfinamide

[0458] [ka]

[0459] Sodium borohydride (101 mg, 2.66 mmol) was added to a solution of (NE)-N-[[(2S)-3,4-dihydro-2H-pyran-2-yl]methylene]-2-methyl-propane-2-sulfinamide (572 mg, 2.66 mmol) in ethanol (10.0 mL) at 0 °C. The ice bath was removed and the reaction stirred for an additional 30 min. The reaction was cooled to 0 °C and sat. NH4Cl (20.0 mL) was added (Caution: Gas evolution). The EtOH was evaporated under reduced pressure and the residue was extracted with EtOAc (30.0 mL). The layers were separated and the organic phase was washed successively with water (10.0 mL) and brine (10.0 mL), dried (Na2SO4), filtered, and concentrated under reduced pressure to give the title compound as a solid (547 mg, 95%). LCMS m / z: ES + [M+H] + (A05) retention time = 2.04 m. This material was used in the following step without further purification.

[0460] Step C. (R)-N-benzyl-N-[[(2S)-3,4-dihydro-2H-pyran-2-yl]methyl]-2-methyl-propane-2-sulfinamide

[0461] [ka]

[0462] Sodium hydride (60%, 116 mg, 2.89 mmol) was added to a mixture of (R)-N-[[(2S)-3,4-dihydro-2H-pyran-2-yl]methyl]-2-methyl-propane-2-sulfinamide (547 mg, 2.52 mmol) and BnBr (448 μL, 3.78 mmol) in DMF (1.5 mL) at 0° C. The mixture was stirred at room temperature for 1 h and then cooled to 0° C., followed by the addition of water (10.0 mL). The aqueous layer was extracted with EtOAc (25.0 mL), and the organic layer was washed successively with water (10.0 mL) and brine (5.0 mL), dried (NaSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (24 g cartridge) using EtOAc and hexanes (0-40%) to give the title compound as an oil (768 mg, 99%). LCMS m / z: ES + [M+H] + :308.18;(A05) Retention time=2.57m.

[0463] Step D. (R)-N-Benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide

[0464] [ka]

[0465] Iodine (272 mg, 1.07 mmol) was added to a suspension of (R)-N-benzyl-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (320 mg, 1.04 mmol) and NaHCO (262 mg, 3.12 mmol) in 1:1 HO / ACN (8.0 mL) at ambient temperature. After 30 min, MeCN was evaporated and the residue was partitioned between EtOAc (20.0 mL) and 1:1 sat. NaHCO / sat. NaSO (10.0 mL). The organic layer was washed with water (5.0 mL) and brine (5.0 mL), dried (Na2SO4), filtered, and concentrated under reduced pressure to give the title compound as a foam (4 diastereoisomers, 450 mg, 96%). LCMS m / z: ES + [M+H] + :452.06; (A05) retention time = 2.30-2.50 m. This material was used in the following reaction without further purification.

[0466] Step E. (R)-N-[[(2S)-5-Azido-6-oxo-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl-propane-2-sulfinamide

[0467] [ka]

[0468] Molecular sieves (4 Å, 500 mg) and pyridinium dichromate (750 mg, 1.99 mmol) were added to a solution of benzyl (R)-N-benzyl-N-[[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]methyl]-2-methyl-propane-2-sulfinamide (450 mg, 997 μmol) in dry DCM (10.0 mL) at ambient temperature under N. After 24 h, the solution was concentrated, and the crude product was partitioned between EtOAc (30.0 mL) and water (15.0 mL). The organic layer was washed with water (2 × 10.0 mL) and brine (10.0 mL), dried (NaSO), and concentrated. The crude product was dissolved in dry DMF (1.5 mL) under N, and NaN (130 mg, 1.99 mmol) was added. After 1 h, the solution was partitioned between EtOAc (30.0 mL) and water (15.0 mL). The organic layer was washed with water (2×10.0 mL) and brine (10.0 mL), dried (NaSO), and concentrated to give the title compound as an oil (235 mg, 64%). LCMS m / z: ES + [M+H] + (A05) retention time = 2.37 m. This material was used in the next reaction without further purification.

[0469] Step F. (R)-N-[[(2S,5R)-5-Azido-6-hydroxy-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl-propane-2-sulfinamide

[0470] [ka]

[0471] A solution of DIBAL-H in toluene (1 M, 1.03 mL, 1.03 mmol) was added dropwise to a solution of (R)-N-[[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl-propane-2-sulfinamide (235 mg, 645 μmol) in dry DCM (8.0 mL) at −78° C. under N2. After 100 min, sat. potassium sodium tartrate (15.0 mL) was slowly added to the solution, followed by water (15.0 mL). The mixture was warmed to room temperature and stirred vigorously for 16 h. The mixture was extracted with DCM (10.0 + 2 × 5.0 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (12 g cartridge) using EtOAc and hexanes (10% to 50%) to give the title compounds as oils (diastereoisomers, 195 mg, 74%). LCMS m / z: ES + [M+H] + :367.19;(A05) Holding time = 2.28~2.40min.

[0472] Step G. Benzyl N-[[(2S,5R)-5-azido-6-hydroxytetrahydropyran-2-yl]methyl]-N-benzyl-carbamate (Ring A1)

[0473] [ka]

[0474] Aqueous HCl (1.0 M, 2.66 mL, 2.66 mmol) was added dropwise to a solution of (R)-N-[[(2S)-5-azido-6-hydroxy-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl-propane-2-sulfinamide (195 mg, 532 μmol) in dioxane (2.7 mL) with vigorous stirring. After 1 h, solid Na2CO3 (451 mg, 4.26 mmol) was added. After an additional 5 min, CbzCl (106 μL, 745 mmol) was added dropwise. After an additional 1 h, dioxane was evaporated, and the residue was partitioned between EtOAc (20.0 mL) and HO (20.0 mL). The organic phase was washed with brine (10.0 mL), dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude material was purified by C18 reverse-phase chromatography (80 g cartridge) using ACN and 0.1% aq. formic acid (40-70%) to give the title compound (mixture of four diastereoisomers) as an oil (167 mg, 79%). UPLC m / z: ES + [M-OH] + UPLC m / z: 379.21; (CSH 5-100% ACN / ammonium formate pH 4), retention times = 2.12, 2.15, 2.20, and 2.22 min. A portion of the material (40 mg) was purified by preparative HPLC (CSH C18 ACN / ammonium formate 50-70%) to give the α-azide as a film (20 mg, 50% recovery). UPLC m / z: (CSH 5-100% ACN / ammonium formate pH 4), retention times = 2.15 and 2.22 min.

[0475] Example 2. Synthesis of benzyl ((1S)-1-((2S,5R)-5-azido-6-hydroxytetrahydro-2H-pyran-2-yl)ethyl) (benzyl)carbamate (Ring A2).

[0476] [ka]

[0477] Step A. ((S)-N-((E)-((S)-3,4-Dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinamide

[0478] [ka]

[0479] (S)-(+)-2-Methyl-2-propanesulfinamide (50 g, 412 mmol) and CuSO (131.7 g, 825 mmol) were mixed in DCM (600 mL). A solution of 2-formyl-3,4-dihydro-2H-pyran (48.6 g, 433 mmol, Synthonix) in DCM (100 mL) was added over 20 min. The mixture was stirred at room temperature for 18 h and then filtered through diatomaceous earth and rinsed with DCM. The filtrate was evaporated under reduced pressure. The material was purified on silica gel by MPLC (5 × 330 g, dry loading) using 0–20% EtO in hexanes as the eluent to give the title compound (23.74 g, 27%, second-eluting diastereoisomer). 1 H NMR (400 MHz, CDCl3) δ 8.08 (d, J = 3.0 Hz, 1H), 6.47 (dt, J = 6.3, 1.6 Hz, 1H), 4.81 - 4.72 (m, 2H), 2.19 - 1.94 (m, 4H), 1.24 (s, 9H).

[0480] Step B. (S)-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide

[0481] [ka]

[0482] (S)—N-((E)-((S)-3,4-Dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinamide (7.30 g, 33.9 mmol) was dissolved in dry DCM (100 mL) and the solution was cooled to −78° C. MeMgBr (3.0 M in EtO, 22.6 mL, 67.8 mmol) was added dropwise to the mixture and stirring was continued at low temperature for 2 h before the mixture was allowed to warm to room temperature. Stirring was continued at room temperature for 1 h. Concentrated aqueous NH4Cl (50 mL) was added and the mixture was extracted with DCM (3 × 50 mL). The combined organic layers were dried (MgSO4) and concentrated in vacuo to give a crude oil containing a mixture of diastereoisomers. The material was purified using silica gel chromatography (120 g cartridge) with hexanes and ethyl acetate (0-100%) to give the title compound as an oil (4.40 g, 56%). LCMS m / z: ES + [M+H] + :232.16.

[0483] Step C. (S)-N-benzyl-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide

[0484] [ka]

[0485] NaH (60% dispersion, 913 mg, 22.8 mmol) was added to a solution of (S)-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide (4.40 g, 19.0 mmol) and benzyl bromide (3.39 mL, 28.5 mmol) in DMF (100 mL) at 0 °C. The mixture was stirred at this temperature for 2 h, and then brine (100 mL) was added at 0 °C. The aqueous layer was extracted with EtO (3 × 50.0 mL). The combined organic layers were dried (MgSO) and concentrated in vacuo to give a residue that was purified using silica gel chromatography (120 g cartridge) using hexanes and ethyl acetate (0–100%). Pure fractions were concentrated in vacuo to give the title compound as an oil (3.50 g, 57%). LCMS m / z:ES + 322.14. 1 H NMR (500 MHz, DMSO) δ 7.45 (d, J = 7.1 Hz, 2H), 7.35 - 7.29 (m, 2H), 7.23 (t, J = 7.3 Hz, 1H), 6.38 (d, J = 6.1 Hz, 1H), 4.65 (ddd, J = 6.2, 4.3, 1.4 Hz, 1H), 4.35 (d, J = 16.5 Hz, 1H), 3.91 (t, J = 11.4 Hz, 1H), 3.80 (td, J = 9.2, 2.3 Hz, 1H), 3.22 - 3.11 (m, 1H), 1.97 - 1.92 (m, 1H), 1.89 - 1.78 (m, 2H), 1.53 - 1.42 (m, 1H), 1.19 (t, J = 3.4 Hz, 3H), 1.10 (s, 8H).

[0486] Step D. (S)-N-benzyl-N-((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide

[0487] [ka] (S)-N-benzyl-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide (4.40 g, 13.7 mmol) and NaHCO3 (3.45 g, 41.1 mmol) were dissolved in HO (75.0 mL) and ACN (75.0 mL). I2 (3.82 g, 15.1 mmol) was added portionwise over 10 min at 0 °C. The cooling bath was removed and stirring was continued at room temperature for 90 min. The reaction mixture was quenched with a saturated aqueous solution of Na2S2O3 (30.0 mL) and extracted with EtOAc (3 x 20.0 mL). The combined organic layers were dried (MgSO4) and concentrated in vacuo to give the title compound as a solid (4.70 g, 74%). LCMS m / z: ES + [M+H] + :466.04.

[0488] Step E. Benzyl ((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)carbamate

[0489] [ka] (S)-N-benzyl-N-((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide (4.70 g, 10.1 mmol) was dissolved in 1,4-dioxane (150 mL) and an aqueous solution of 1 N HCl (25.2 mL, 25.2 mmol) was added. Stirring was continued at room temperature for 20 minutes, after which solid Na2CO3 (8.56 g, 80.8 mmol) was added to the mixture. Stirring was continued for 20 minutes, and then CbzCl (1.72 mL, 12.1 mmol) was added dropwise. Stirring was continued for 2 hours. Water (200 mL) was added and the mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were dried (MgSO4) and concentrated in vacuo to give a residue that was purified using silica gel chromatography (80 g cartridge) using hexanes and ethyl acetate (0-100%). Pure fractions were concentrated in vacuo to give the title compound as an oil (3.70 g, 74%). LCMS m / z: ES + [M+H] + :518.04.

[0490] Step F. Benzyl ((1S)-1-((2S)-5-azido-6-oxotetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate

[0491] [ka] Benzyl benzyl ((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)carbamate (3.70 g, 7.47 mmol) was dissolved in DCM (350 mL). 4 Å molecular sieves (1.00 g) were suspended in the mixture, and PDC (8.43 g, 22.4 mmol) was added. Stirring was continued for 24 h at room temperature, followed by filtration through a pad of diatomaceous earth using ethyl acetate and concentration in vacuo to give crude benzyl benzyl ((1S)-1-((2S)-5-iodo-6-oxotetrahydro-2H-pyran-2-yl)ethyl)carbamate as an oil. The crude product was dissolved in DMF (50.0 mL). NaN (971 mg, 14.9 mmol) was added as a solid, and stirring was continued for 2 h at room temperature. The solvent was removed in vacuo and the residue was dissolved in EtOAc (100 mL) and washed with water (3 x 25 mL). The organic layer was dried (MgSO4) and concentrated in vacuo to give an oil which was purified using silica gel chromatography (80 g cartridge) with ethyl acetate and hexanes (0-100%). Pure fractions were combined and concentrated in vacuo to give the title compound as an oil (1.40 g, 46%). LCMS m / z: ES + [M+Na] + :431.13;calc:431.18.

[0492] Step G. Benzyl ((1S)-1-((2S,5R)-5-azido-6-hydroxytetrahydro-2H-pyran-2-yl)ethyl) (benzyl)carbamate (Ring A2)

[0493] [ka] Benzyl ((1S)-1-((2S)-5-azido-6-oxotetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (1.40 g, 3.43 mmol) was dissolved in dry DCM (75.0 mL), and the mixture was cooled to −78 °C. A solution of DIBAL-H (1.0 M in toluene, 6.86 mL, 6.86 mmol) was added to the mixture, and stirring was continued at low temperature for 1 h. EtOH (2.00 mL) was added dropwise to the cold reaction mixture, which was then poured into a saturated aqueous solution of Rochelle's salt (200 mL). The mixture was stirred vigorously for 1 h and then extracted with EtOAc (3 × 50.0 mL). The combined organic layers were dried (MgSO4) and concentrated in vacuo to give a residue that was purified using silica gel chromatography (80 g cartridge) with hexanes and ethyl acetate (0–70%). The pure fractions were collected and concentrated in vacuo to give the title compound as a mixture of diastereoisomers (800 mg, 57%). LCMS m / z: ES + [M+Na] + :433.16;calc:433.19.

[0494] Example 3. Synthesis of benzyl N-[(1R)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (Ring A3).

[0495] [ka]

[0496] Step A. (R)-N-((E)-((S)-3,4-Dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinamide

[0497] [ka]

[0498] (R)-(+)-2-Methyl-2-propanesulfinamide (81.1 g, 669 mmol) and CuSO (213.5 g, 1.34 mol) were mixed in DCM (700 mL). A solution of 2-formyl-3,4-dihydro-2H-pyran (75 g, 669 mmol) in DCM (100 mL) was added over 20 min. The mixture was stirred at room temperature for 72 h. The mixture was filtered through diatomaceous earth and rinsed with DCM. The filtrate was evaporated under reduced pressure. The material was purified on silica gel (5 × 330 g, dry loading) by MPLC using 0–10% EtO in hexanes to give the title compound (28 g, 19%) as a solid. 1 H NMR (400 MHz, CDCl3) δ 8.07 (d, J = 3.1 Hz, 1H), 6.40 (d, J = 5.8 Hz, 1H), 4.76 - 4.68 (m, 1H), 4.68 - 4.63 (m, 1H), 2.13 - 1.83 (m, 4H), 1.18 (s, 9H).

[0499] Step BN-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide

[0500] [ka]

[0501] MeMgBr (3.0 M in EtO, 22.0 mL, 66.0 mmol) was added to a solution of (R)-N-((E)-((S)-3,4-dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinamide (7.10 g, 33.0 mmol) in dry DCM (110 mL) at −40° C. under N. After 2 h, the reaction was allowed to warm to room temperature within 1 h. After a further 14 h, sat. NH Cl (160 mL) was added dropwise (CAUTION: gas evolution). The two phases were separated, and the aqueous phase was extracted with DCM (3 × 20.0 mL). The combined organic layers were dried (NaSO), filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (220 g cartridge) with EtOAc and hexanes (20-45%) to afford the title compound as a solid (6.00 g, 79%). 1 H NMR (500 MHz, CDCl3) δ 6.37 (d, J = 6.1 Hz, 1H), 4.75 - 4.61 (m, 1H), 3.93 (ddd, J = 11.2, 3.3, 2.0 Hz, 1H), 3.69 (d, J = 6.1 Hz, 1H), 3.56 (pd, J = 6.7, 3.5 Hz, 1H), 2.16 - 2.06 (m, 1H), 1.99 (dtt, J = 17.1, 5.6, 1.6 Hz, 1H), 1.81 (ddd, J = 13.3, 6.6, 1.8 Hz, 1H), 1.66 (dtd, J = 13.3, 11.4, 5.8 Hz, 1H), 1.22 (s, 9H), 1.18 (d, J = 6.7 Hz, 3H).

[0502] Step C. (R)-N-Benzyl-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide

[0503] [ka]

[0504] NaH (60%, 187 mg, 4.89 mmol) was added to a mixture of (R)-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (1.03 g, 4.44 mmol) and BnBr (0.79 mL, 6.67 mmol) in DMF (60 mL) at 0 °C. The mixture was stirred at room temperature for 1 h, and then brine (250 mL) was added at 0 °C. The aqueous layer was extracted with EtO (3 × 80 mL). The combined organic layers were dried over MgSO and concentrated under reduced pressure. The material was purified on silica gel (40 g, dry loading) by MPLC using hexanes to 60% EtOAc in hexanes to give the title compound (1.2 g, 84%) as an oil. 1 H NMR (400 MHz, cdcl3) δ 7.37 (d, J = 7.5 Hz, 2H), 7.32 (t, J = 7.4 Hz, 2H), 7.26 (d, J = 3.8 Hz, 1H), 6.32 (d, J = 6.2 Hz, 1H), 4.64 (s, 1H), 4.39 (d, J = 15.3 Hz, 1H), 4.14 (d, J = 15.2 Hz,1H), 3.66 - 3.57 (m, 1H), 3.32 - 3.20 (m, 1H), 1.98 - 1.80 (m, 3H), 1.54 - 1.43 (m, 1H), 1.37 (d, J = 6.8 Hz, 3H), 1.19 (s, 9H).

[0505] Step D. (R)-N-Benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide

[0506] [ka]

[0507] I2 (947 mg, 3.73 mmol) was added portionwise to a suspension of (R)-N-benzyl-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (1.2 g, 3.73 mmol) and NaHCO3 (941 mg, 11.2 mmol) in ACN (50 mL) and HO (50 mL) at 0 °C. The mixture was stirred at room temperature for 1 h, then a saturated aqueous solution of Na2SO3 (10 mL) was added. The aqueous layer was extracted with EtOAc (3 × 150 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated under reduced pressure to give the title compound (1.6 g, 92%) as a solid. M+H + :466.1.

[0508] Step E. Benzyl N-benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]carbamate

[0509] [ka]

[0510] 1N HCl (6.45 mL, 6.45 mmol) was added to (R)-N-benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (1.2 g, 2.58 mmol) in dioxane (40 mL). The mixture was stirred at room temperature for 20 min, and then Na2CO3 (2.19 g, 20.6 mmol) was added. After 20 min, CbzCl (0.51 mL, 3.61 mmol) was added dropwise. The mixture was stirred at room temperature for 2 h. Water (200 mL) was added. The separated aqueous layer was extracted with EtOAc (3 × 150 mL). The combined organic layers were washed with brine, dried over MgSO4, and concentrated under reduced pressure. The material was purified by MPLC on silica gel (80 g, dry loading) using hexanes to 80% EtOAc to give the title compound (1.1 g, 86%) as a solid. + :496.0.

[0511] Step F. Benzyl N-[(1R)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate

[0512] [ka]

[0513] Dess-Martin periodinane (1.88 g, 4.44 mmol) was added to a solution of benzyl N-benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]carbamate (1.10 g, 2.22 mmol) in DCM (100 mL) at 0 °C. The mixture was stirred at room temperature for 5 h. Water (100 mL) was added, followed by a saturated aqueous solution of NaSO. The separated aqueous layer was extracted with DCM (2 × 50 mL). The combined organic layers were washed with saturated aqueous NaHCO (2 × 100 mL), brine (100 mL), dried over MgSO, and concentrated under reduced pressure. The residue was taken up in anhydrous DMF (75 mL), and NaN (217 mg, 3.33 mmol) was added. The mixture was stirred at room temperature for 15 min, then brine (300 mL) was added. The aqueous layer was extracted with EtO (3 x 100 mL). The combined organic layers were dried over MgSO and concentrated under reduced pressure. The material was purified on silica gel (40 g, dry loading) by MPLC using hexanes to EtOAc to give the title compound (800 mg, 88%) as an oil. M+H + :409.3.

[0514] Step G. Benzyl N-[(1R)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (Ring A3)

[0515] [ka]

[0516] A solution of DIBAL-H (1 M in toluene, 11.8 mL, 11.8 mmol) was added dropwise to a solution of benzyl N-[(1R)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (800 mg, 1.96 mmol) in DCM (60 mL) at −78° C. After 1 h, EtOH (0.5 mL) was added dropwise at −78° C. The mixture was poured into a saturated aqueous solution of Rochelle's salt (300 mL). The mixture was stirred vigorously for 1 h. The separated aqueous layer was extracted with DCM (2×75 mL). The combined organic layers were washed with brine, dried over MgSO4, and concentrated under reduced pressure. The residue was purified on silica gel (40 g, dry loading) by MPLC using hexanes to 60% EtOAc in hexanes to give the title compound (363 mg, 41%) as an oil. + :411.2.

[0517] Example 4. Synthesis of (1R,2R,3S,4R,5S,6S)-3,5-bis(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexane-1,2,4-triyl triacetate (Ring B1).

[0518] [ka]

[0519] Step A: Synthesis of (1S,2R,3S,4S,5R,6S)-3,5-diamino-6-(((2R,3R,4R,5S)-3-(((2R,3S,4S,5R,6S)-4,5-dihydroxy-6-(hydroxymethyl)-3-(methylamino)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-4-(hydroxymethyl)-5-methyltetrahydrofuran-2-yl)oxy)cyclohexane-1,2,4-triol

[0520] [ka]

[0521] Dihydrostreptomycin sulfate (50 g), Ba(OH)-8H0 (4 equivalents), and water (500 mL) were combined and heated to reflux for 2 days. After cooling to room temperature, CO was bubbled through the reaction until it was slightly acidic (pH 6-7). The resulting mixture was filtered through diatomaceous earth and washed twice with water (75 mL). The aqueous layer was concentrated in vacuo, and the aqueous solution was used directly without further purification.

[0522] Step B: Synthesis of dibenzyl ((1S,2S,3R,4S,5S,6R)-4-(((2R,3R,4R,5S)-3-(((2R,3S,4S,5R,6S)-3-(((benzyloxy)carbonyl)(methyl)amino)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-4-(hydroxymethyl)-5-methyltetrahydrofuran-2-yl)oxy)-2,5,6-trihydroxycyclohexane-1,3-diyl)dicarbamate

[0523] [ka]

[0524] To the crude compound from Step A was added Cbz-OSu (57.7 g, 3.4 eq) in THF (110 mL) over approximately 1 h at room temperature. The reaction was stirred at room temperature for 2 days. Saturated aqueous sodium bicarbonate (50 mL) was added, and the product was extracted into EtOAc (2 × 75 mL) and washed with brine (50 mL). The combined organics were dried over sodium sulfate, and the solvent was removed in vacuo. The crude product was dissolved in 2-butanone (80 ml) and slowly added to MTBE (750 mL). The precipitated solid was collected by filtration and dried in a vacuum oven at 50 °C to give the title product (52.1 g) as an off-white solid (72% pure by absorbance at 210 nm).

[0525] Step C: Synthesis of (1R,2R,3S,4R,5R,6S)-6-(((2R,3R,4R,5S)-4-(acetoxymethyl)-3-(((2R,3S,4S,5R,6S)-4,5-diacetoxy-6-(acetoxymethyl)-3-(((benzyloxy)carbonyl)(methyl)amino)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-5-methyltetrahydrofuran-2-yl)oxy)-3,5-bis(((benzyloxy)carbonyl)amino)cyclohexane-1,2,4-triyl triacetate

[0526] [ka]

[0527] The crude compound from Step B (52.1 g) was dissolved in EtOAc (210 mL) and DMAP (721 mg, 0.1 eq) was added at room temperature. DIPEA (103 mL) and AcO (51 mL) in EtOAc (80 mL) were added dropwise to the reaction mixture over 1 h at room temperature. The reaction was stirred overnight at room temperature. The reaction was transferred to a separatory funnel with EtOAc (50 mL, 0.9 V) and washed with water (50 mL), sodium bicarbonate solution (2 x 50 mL), and brine (50 mL). The organics were dried over sodium sulfate and filtered to give the title product as a cream-colored foam (70 g; 63% AP at 210 nm).

[0528] Step D: Synthesis of (1R,2R,3S,4R,5S,6S)-3,5-bis(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexane-1,2,4-triyl triacetate (Ring B1)

[0529] [ka]

[0530] The crude product from Step C (53 g) was added to DCM (420 mL), dodecanethiol (29.4 mL, 2.1 eq), and BF3·Et2O (16.3 mL, 2.2 eq). The reaction was heated to reflux for 2 h. The reaction was washed with aqueous sodium bicarbonate, water, and brine (20 mL each). The organic portion was concentrated in vacuo, and the resulting solid was stirred vigorously with MeCN / hexane (1:2, 210 mL) to give the title product. The crude product was triturated with heptane (300 mL) to give the title product (23.9 g, 71% yield).

[0531] Example 5. Synthesis of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid (F-HABA).

[0532] [ka]

[0533] Step A: Synthesis of methyl (R)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyacetate.

[0534] [ka]

[0535] To a solution of D-isoascorbic acid (100 g, 0.568 mol, 1.0 eq.) and 2,2-dimethoxypropane (76.8 mL, 0.624 mol, 1.1 eq.) in dry acetone (500 mL), concentrated H2SO4 (0.6 mL, 11.2 mmol) was slowly added, and the mixture was stirred at 0 °C for 5 h. NaHCO3 (142.8 g, 1.7 mol, 3.0 eq.) and H2O (400 mL) were slowly added to the reaction mixture. Acetone was removed under reduced pressure using a rotary evaporator. 30% H2O2 (128.4 mL, 1.132 mol, 2.0 eq.) was slowly added to the residue using a dropping funnel at 0 °C. After stirring the mixture for 3 h, Na2SO3 (14.28 g, 113.2 mmol, 0.2 eq.) was added, and the mixture was stirred for 30 min. Sodium bicarbonate (192.8 g, 2.28 mol, 4.0 eq.) and dimethyl sulfate (225.6 mL, 2.36 mol, 4.0 eq.) were added, and the mixture was stirred at 50 °C for 5 h. The reaction mixture was filtered, and the aqueous layer was extracted with CHCl (800 mL × 2). The organic layers were combined, dried over anhydrous MgSO, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography eluting with petroleum ether / ethyl acetate (2:1, v / v) to give methyl (R)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyacetate (70 g, 64.8%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 4.35-4.14 (m, 2H), 4.01-3.90 (m, 2H), 3.75 (s, 3H), 3.03 (d, J = 6.4 Hz, 1H), 1.36 (s, 3H), 1.29 (s, 3H).

[0536] Step B: Synthesis of (S)-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethane-1,2-diol.

[0537] [ka]

[0538] To a suspension of LAH (16.59 g, 0.44 mol, 1.2 eq.) in dry THF (600 mL) was added dropwise a solution of methyl (R)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyacetate (70 g, 0.36 mol, 1.0 eq.) in THF (200 mL) under N. The resulting mixture was stirred at room temperature for 3 h, and then HO (20 mL) was added dropwise, followed by 15% aq. NaOH (20 mL) and HO (60 mL). The reaction mixture was filtered and concentrated, and the residue was purified by flash column chromatography eluting with petroleum ether / ethyl acetate (1:1) to give (S)-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethane-1,2-diol (45 g, 75.4% yield). 1 H NMR (400 MHz, DMSO-d6) δ 4.21 (br, s, 2 H), 3.79-3.95 (m, 3 H), 3.29-3.50 (m, 3H), 1.30 (s, 3 H), 1.25 (s, 3 H).

[0539] Step C: Synthesis of (S)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyethyl 4-methylbenzenesulfonate.

[0540] [ka]

[0541] To a solution of (S)-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethane-1,2-diol (45 g, 277.8 mmol, 1.0 eq.) in dry pyridine (500 mL) was added TsCl (52.96 g, 277.8 mmol, 1.0 eq.) portionwise over 1 h at 0 °C. The resulting mixture was stirred overnight at room temperature, and then the solvent was removed. The residue was dissolved in EA (1000 mL), washed with saturated aqueous NH Cl (500 mL × 2), dried over MgSO, filtered, and concentrated. The residue was purified by flash column chromatography eluting with petroleum ether / ethyl acetate (4:1) to give (S)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyethyl 4-methylbenzenesulfonate (59 g, 75.9% yield). 1 H NMR (400 MHz, DMSO) δ 7.79 (d, J = 8.2 Hz, 2H), 7.49 (d, J = 8.1 Hz, 2H), 5.53 (d, J = 5.9 Hz, 1H), 4.11 - 4.03 (m, 1H), 3.98 - 3.85 (m, 2H), 3.80 (td, J = 10.0, 4.9 Hz, 2H), 3.56 - 3.44 (m, 1H), 2.42 (s, 3H), 1.21 (s, 3H), 1.19 (s, 3H).

[0542] Step D: Synthesis of (S)-2-azido-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethan-1-ol.

[0543] [ka]

[0544] To a solution of (S)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyethyl 4-methylbenzenesulfonate (59 g, 186.6 mmol, 1.0 eq.) in dry DMF (500 mL) was added NaN (36.38 g, 559.7 mmol, 3.0 eq.) at rt. The mixture was warmed to 80 °C and stirred overnight. The resulting mixture was poured into saturated aqueous NaCO (2000 mL) and extracted with ethyl acetate (1000 mL × 2). The combined organic phase was washed with saturated aqueous NH Cl (1000 mL × 2), dried over MgSO, filtered, and concentrated. The residue was purified by flash column chromatography eluting with petroleum ether / ethyl acetate (8:1) to give (S)-2-azido-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethan-1-ol (34 g, 97.4% yield). 1 H NMR (400 MHz, DMSO) δ 5.54 (d, J = 5.8 Hz, 1H), 3.99 (dd, J = 7.7, 5.9 Hz, 1H), 3.92 - 3.79 (m, 2H), 3.53 (ddd, J = 8.3, 6.5, 2.8 Hz, 1H), 3.32 (m, 1H), 3.22 (dd, J = 12.8, 6.5 Hz, 1H), 1.30 (s, 3H), 1.25 (s, 3H).

[0545] Step E: Synthesis of (R)-4-((R)-2-azido-1-fluoroethyl)-2,2-dimethyl-1,3-dioxolane.

[0546] [ka]

[0547] To a solution of (S)-2-azido-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethan-1-ol (34 g, 0.18 mol, 1.0 eq) in dry DCM (170 mL) was added a solution of DAST (73 mL, 0.54 mol, 3.0 eq) dropwise at −20° C., and the reaction mixture was then warmed to rt and stirred overnight. The resulting mixture was slowly poured into saturated aqueous NaHCO (1500 mL) and extracted with DCM (500 mL × 2). The combined organic phases were dried over MgSO, filtered, and concentrated. The residue was purified by flash column chromatography eluting with petroleum ether / ethyl acetate (30:1) to give (R)-4-((R)-2-azido-1-fluoroethyl)-2,2-dimethyl-1,3-dioxolane (3.4 g, 9.9% yield). 1 H NMR (400 MHz, CDCl3) δ 4.57 (ddt, J = 48.0, 7.4, 3.6 Hz, 1H), 4.24 (dtd, J = 21.4, 6.7, 3.6 Hz, 1H), 4.14 - 4.05 (m, 1H), 3.94 (dd, J = 8.4, 6.5 Hz, 1H), 3.63 (ddd, J = 17.0, 13.5, 7.5 Hz, 1H), 3.44 (ddd, J = 27.4, 13.5, 3.6 Hz, 1H), 1.44 (s, 3H), 1.38 (s, 3H). 19 F NMR (400 MHz, CDCl3) δ 198.91 (s, 1 F).

[0548] Step F: Synthesis of (2R,3R)-4-azido-3-fluorobutane-1,2-diol.

[0549] [ka]

[0550] To a solution of (R)-4-((R)-2-azido-1-fluoroethyl)-2,2-dimethyl-1,3-dioxolane (3.4 g, 18.0 mmol, 1.0 eq.) in THF (30 mL) was added 10% HCl (4 mL) dropwise at 0 °C. The mixture was warmed to rt and stirred overnight. The organic solvent was removed, and the residue was basified to pH = 9 with saturated aqueous Na2CO3, extracted with EA (40 mL × 4), dried over MgSO4, filtered, and concentrated. The residue was purified by flash column chromatography eluting with petroleum ether / ethyl acetate (1:1) to give (2R,3R)-4-azido-3-fluorobutane-1,2-diol (2.5 g, yield: 93.3%). 1 H NMR (400 MHz, DMSO) δ 5.17 (d, J = 5.9 Hz, 1H), 4.86 (t, J = 5.5 Hz, 1H), 4.74 (ddt, J = 48.8, 8.4, 2.9 Hz, 1H), 3.79 - 3.44 (m, 5H). 19 F NMR (400 MHz, DMSO-d6) δ 202.16 (s, 1 F).

[0551] Step G: Synthesis of (2R,3R)-4-azido-1-((tert-butyldimethylsilyl)oxy)-3-fluorobutan-2-ol.

[0552] [ka]

[0553] To a solution of (2R,3R)-4-azido-3-fluorobutane-1,2-diol (2.5 g, 16.7 mmol, 1.0 eq.) in DCM (50 mL) was added DMAP (4.51 g, 36.9 mmol, 2.2 eq.) followed by TBS-Cl (2.77 g, 18.5 mmol, 1.1 eq.) in small portions at 0 °C. The reaction mixture was warmed to rt and stirred for 3 days. The organic solvent was evaporated, and the residue was purified by flash chromatography eluting with petroleum ether / ethyl acetate (10:1) to give (2R,3R)-4-azido-1-((tert-butyldimethylsilyl)oxy)-3-fluorobutan-2-ol (2.6 g, yield: 59.1%). 1 H NMR (400 MHz, DMSO) δ 5.17 (d, J = 5.7 Hz, 1H), 4.71 - 4.50 (m, 1H), 3.73 - 3.37 (m, 5H), 0.82 (s, 9H), 0.00 (s, 6H). 19 F NMR (400 MHz, DMSO-d6) δ 203.48 (s, 1 F).

[0554] Step H: Synthesis of ((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butoxy)(tert-butyl)dimethylsilane.

[0555] [ka]

[0556] To a solution of (2R,3R)-4-azido-1-((tert-butyldimethylsilyl)oxy)-3-fluorobutan-2-ol (2.6 g, 9.88 mmol, 1.0 eq.) in THF (250 mL) was added NaH (474 ​​mg, 11.86 mmol, 1.2 eq., 60%) in portions at 0 °C. After stirring at 0 °C for 30 min, TBAI (3.64 g, 9.88 mmol, 1.0 eq.) was added, followed by the dropwise addition of PMB-Cl (2.0 mL, 14.73 mmol, 1.5 eq.) at 0 °C. The reaction was then warmed to rt and stirred overnight. The mixture was poured into saturated aqueous NH Cl and extracted with EA (50 mL × 3). The extract was dried over MgSO, filtered, and concentrated. The residue was purified by flash chromatography eluting with petroleum ether / ethyl acetate (40:1) to give ((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butoxy)(tert-butyl)dimethylsilane (2.5 g, yield: 66.1%). 1 H NMR (400 MHz, dioxane) δ 8.31 (d, J = 8.5 Hz, 2H), 7.95 (d, J = 8.6 Hz, 2H), 5.90 - 5.69 (m, 1H), 5.58 (dd, J = 40.1, 11.2 Hz, 2H), 4.79 (s, 3H), 4.78 - 4.48 (m, 5H), 1.91 (s, 9H), 1.09 (s, 6H). 19 F NMR (400 MHz, DMSO-d6) δ 199.15 (s, 1 F).

[0557] Step I: Synthesis of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butan-1-ol.

[0558] [ka]

[0559] To a solution of ((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butoxy)(tert-butyl)dimethylsilane (2.5 g, 6.48 mmol, 1.0 eq.) in DMSO (15 mL) was added CsF (2.98 g, 19.44 mmol, 3.0 eq.), followed by 5 drops of MeOH. The mixture was stirred at rt overnight, then poured into saturated NaCl (40 mL) and extracted with EA (40 mL × 3). The extract was dried over MgSO, filtered, and concentrated. The residue was purified by flash chromatography eluting with petroleum ether / ethyl acetate (4:1) to give (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butan-1-ol (1 g, yield: 56.8%). 1 H NMR (400 MHz, DMSO) δ 7.35 - 7.20 (m, 2H), 7.01 - 6.83 (m, 2H), 5.76 (s, 3H), 4.93 (t, J = 5.3 Hz, 1H), 4.74 (ddd, J = 25.8, 8.0, 5.6 Hz, 1H), 4.52 (dd, J = 49.2, 11.3 Hz, 2H), 3.74 (s, 3H), 3.69 (ddd, J = 18.8, 13.9, 8.1 Hz, 1H), 3.58 - 3.42 (m, 4H). 19 F NMR (400 MHz, DMSO-d6) δ 199.45 (s, 1 F).

[0560] Step J: Synthesis of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid (F-HABA).

[0561] [ka]

[0562] To a solution of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butan-1-ol (1 g, 3.76 mmol, 1.0 eq.) in ACN / HO (10 mL / 10 mL) was added PhI(OAc) (2.99 g, 9.41 mmol, 2.5 eq.), followed by TEMPO (292 mg, 1.88 mmol, 0.5 eq.). The mixture was stirred overnight at rt. Sodium hydroxide (2 N, 10 mL) was added, and the mixture was extracted with PE (20 mL × 4), and the extract was discarded. The aqueous phase was acidified to a pH of 3 with 2 N HCl and extracted with EtOAc (30 mL × 4). The extract was dried over NaSO and concentrated. The residue was purified by flash chromatography eluting with DCM / MeOH (20:1, v / v) to give (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid (500 mg, yield: 47.5%). 1 H NMR (400 MHz, DMSO) δ 7.30 (d, J = 8.7 Hz, 2H), 6.93 (d, J = 8.7 Hz, 2H), 5.11 - 4.86 (m, 1H), 4.52 (dd, J = 119.7, 11.0 Hz, 2H), 4.19 (dd, J = 31.5, 2.6 Hz, 1H), 3.76 (s, 3H), 3.75 - 3.65 (m, 2H). 19 F NMR (400 MHz, DMSO-d6) δ 196.81 (s, 1 F)

[0563] Example 6. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (Compound 1).

[0564] [ka]

[0565] Step A: Synthesis of (3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl(E)-2,2,2-trifluoro-N-phenylacetimidate

[0566] [ka]

[0567] A mixture of benzyl N-[[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]methyl]-N-benzyl-carbamate (Example 1, 1.1 g, 2.50 mmol), CsCO (2.44 g, 7.49 mmol), and (1E)-2,2,2-trifluoro-N-phenyl-acetimidoyl chloride (1.2 mL, 7.49 mmol) in DCM (20 mL) was stirred at rt for 18 h. The mixture was filtered, rinsed with DCM, and concentrated under reduced pressure. The material was purified by silica gel chromatography (80 g cartridge) using a gradient of 0-30% EtOAc in hexanes to give the title product (1.2 g, 85%) as a syrup. m / z (ESI) [M+Na] + 590.3, HPLC(A05)tR=3.02min.

[0568] Step B: Synthesis of (1S,2S,3R,4S,5S,6R)-6-(((3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-3,5-bis(((benzyloxy)carbonyl)amino)cyclohexane-1,2,4-triyl triacetate

[0569] [ka]

[0570] [(3R,6S)-3-Azido-6-[[benzyl(benzyloxycarbonyl)amino]methyl]tetrahydropyran-2-yl](1E)-2,2,2-trifluoro-N-phenyl-ethaneimidate (1.2 g, 2.12 mmol) and [(1S,2R,3S,4S,5R,6R)-3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (Ring B1, Example 4, 1.816 g, 3.17 mmol) were coevaporated with anhydrous toluene (2 × 5 mL). The mixture was dried under reduced pressure for 30 min. The residue was taken up in anhydrous DCM (100 mL) and then transferred via cannula to a RBF containing pre-activated 3 Å and 4 Å sieves. The mixture was stirred at rt for 20 min and then cooled to −25 °C. Bi(OTf)3 (1.526 g, 2.32 mmol) was added in one portion, and the mixture was stirred at -25 °C for 20 min and at rt for 2 h. The mixture was diluted with saturated aqueous NH4Cl (50 mL) and DCM (100 mL) and then filtered (diatomaceous earth). The separated aqueous layer was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (20 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse-phase chromatography using a gradient of 5 to 100% ammonium formate (10 mM, pH 4) in MeCN and lyophilized to afford the title product (1.22 g, 59%) as an anomeric mixture (α / β = ca. 2.7:1). m / z (ESI) [M+H] + :951.5, HPLC tR=2.82 min.

[0571] Step C: Synthesis of dibenzyl ((1R,2R,3S,4R,5R,6S)-4-(((3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2,5,6-trihydroxycyclohexane-1,3-diyl)dicarbamate

[0572] [ka]

[0573] To a suspension of [(1S,2R,3S,4S,5R,6S)-3,4-diacetoxy-5-[(3R,6S)-3-azido-6-[[benzyl(benzyloxycarbonyl)amino]methyl]tetrahydropyran-2-yl]oxy-2,6-bis(benzyloxycarbonyl-amino)cyclohexyl]acetate (1.22 g, 1.28 mmol) in MeOH (30 mL) was added KCO (887 mg, 6.41 mmol). The suspension was stirred at rt for 80 min and then filtered. The filtrate was concentrated under reduced pressure (bath temperature less than 30 °C) to give a residue. Water (15 mL) was added to form a precipitate, which was sonicated, filtered, and washed with water (50 mL). The solid was collected, dissolved in DCM / MeOH (50 mL, 10:1), dried (MgSO), filtered, and concentrated under reduced pressure to give the title compound (1.06 g, crude, quant.) as a solid. m / z (ESI) [M+H] + ;825.4, HPLC A(05)tR=2.63min.

[0574] Step D: Synthesis of dibenzyl((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate

[0575] [ka]

[0576] A mixture of benzyl N-[[(2S,5R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxycyclohexoxy]tetrahydropyran-2-yl]methyl]-N-benzylcarbamate (1 g, 1.21 mmol), 1,1-dimethoxycyclohexane (1.11 mL, 7.27 mmol), and PPTS (122 mg, 0.485 mmol, dried on V10 via high boiling method before use) in dry DME (30 mL) was stirred at 90 °C for 2 h. The mixture was cooled and then concentrated under reduced pressure. The material was purified by reverse phase column in ACCQ using 65-87% ammonium formate in MeCN, pH 4 to give the title compound as a solid (633 mg, 58%, α anomer) and β anomer (250 mg, 23%). m / z: ES+ [M+H] + 905.1; LCMS (A05); tR = 2.99 min (α anomer).

[0577] Step E: Synthesis of dibenzyl((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxole-2,1′-cyclohexane]-5,7-diyl)dicarbamate

[0578] [ka]

[0579] NaH (60% oil dispersion, 64.5 mg, 1.62 mmol) was added to a mixture of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1′-cyclohexane]-5,7-diyl)dicarbamate (630 mg, 0.644 mmol) and BnBr (0.94 mL, 7.85 mmol) in THF (20 mL) at 0° C. under nitrogen. The mixture was stirred at 0° C. for 3 h, then AcOH (0.1 mL, 1.29 mmol) and EtOAc (30 mL) were added. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (40 g cartridge) using a gradient of 0-50% EtOAc in hexanes to give the title compound as a solid (630 mg, 91%). m / z: ES+ [M+H] + 995.5; LCMS(A05); tR=3.13 min.

[0580] Step F: Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate

[0581] [ka]

[0582] A solution of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxole-2,1′-cyclohexane]-5,7-diyl)dicarbamate (630 mg, 0.633 mmol) in THF (12 mL), water (3.5 mL), and AcOH (8 mL) was heated at 80° C. for 9 h and then cooled to rt. The mixture was diluted at 0° C. by slow addition of saturated aqueous NaHCO (ca. 50 mL) and stirred for 20 min. The separated aqueous layer was extracted with EtOAc (3×50 mL). The combined organic layers were washed with saturated aqueous NaHCO3 (3 x 10 mL), water (3 x 10 mL), brine (2 x 10 mL), dried (MgSO4), filtered, and concentrated under reduced pressure to give the title product (570 mg, 98%) as an off-white solid. This material was used directly without further purification. m / z (ESI) [M+H] + :915.4, HPLC(A05)tR=2.85min.

[0583] Step G: Synthesis of benzyl (((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)methyl)(benzyl)carbamate (Ring AB1)

[0584] [ka]

[0585] Sodium hydride (60% oil dispersion, 125 mg, 3.11 mmol) was added to a solution of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate (570 mg, 0.623 mmol) in dry DMF (9 mL) at 0° C. The mixture was stirred at 0° C. for 3 h, then diluted with AcOH (0.28 mL, 4.83 mmol) and concentrated under reduced pressure. The material was purified on silica gel (40 g) by MPLC using a gradient of 0-100% EtOAc in hexanes to give the title compound (440 mg, 88% yield) as a solid. m / z (ESI) [M+H] + :807.4, HPLC(A05)tR=2.77min.

[0586] Step H: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((3aR,4S,5S,6R,7S,7aS)-6-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-oxooctahydrobenzo[d]oxazol-7-yl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0587] [ka]

[0588] To a mixture of [(2R,3R,4R)-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-5-hydroxy-tetrahydrofuran-2-yl]methyl acetate (Ring CD1, WO 2019194858, incorporated by reference; 1157 mg, 2.18 mmol) and KCO (452.4 mg, 3.272 mmol) in dry DCM (15 mL) was added CClCN (0.328 mL, 3.27 mmol). The mixture was stirred at rt for 18 h, then filtered over diatomaceous earth, rinsed with CHCl, and concentrated under reduced pressure. Benzyl N-[[(2S,5R,6R)-6-[[(3aR,4S,5S,6R,7S,7aS)-4-benzyloxy-5-(benzyloxycarbonylamino)-7-hydroxy-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-6-yl]oxy]-5-azido-tetrahydropyran-2-yl]methyl]-N-benzylcarbamate (Ring AB1, 440 mg, 0.545 mmol) in dry DCM (30 mL) was added to the crude material and then transferred to a RBF containing preactivated 3 Å and 4 Å sieves. The mixture was cooled to −78 °C, and then BF3°OEt2 (1.02 mL, 8.23 ​​mmol) was added dropwise at −78 °C. Upon completion of the addition, the dry ice bath was immediately removed. The mixture was stirred at rt for 1 h and then diluted with a saturated aqueous solution of NaHCO3 (15 mL). The mixture was filtered through diatomaceous earth, and the solid was rinsed with DCM (200 mL). The separated aqueous layer was extracted with DCM (3 x 20 mL). The combined organic layers were washed with brine (15 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse-phase column (ACCQ) using a gradient of 64% to 85% MeCN in ammonium formate (10 mM, pH 4) to give the title product (455 mg, 63%). m / z (ESI) [M+H] + :1319.5, HPLC(A05)tR=2.95min.

[0589] Step I: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-amino-6-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0590] [ka]

[0591] LiOH·H2O (434 mg, 10.35 mmol) was dissolved in [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6[[benzyl(benzyloxycarbonyl)amino]methyl]-tetrahydropyran-2-yl]oxy-4-benzyloxy-5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7, To a solution of 7a-hexahydro-3H-1,3-benzoxazol-7-yl]oxy]-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-tetrahydrofuran-2-yl]methyl acetate (455 mg, 0.345 mmol) in 1,4-dioxane (12 mL) and water (6 mL) was added. The mixture was stirred at 40° C. for 18 h, then cooled to rt and diluted with EtOAc (60 mL). The separated aqueous layer was extracted with EtOAc (3×20 mL). The combined organic layers were washed with water (3×5 mL), brine (10 mL), dried (MgSO), filtered, and concentrated under reduced pressure to give the title compound (382 mg, crude, 98%) as an off-white solid. The material was used directly without further purification. m / z(ESI)[M+H] +:1125.5, HPLC(A05)tR=2.43min.

[0592] Step J: (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanamide)-6-(((2R,3R,6S)-3-azido-6-((benzyl( Synthesis of (benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0593] [ka]

[0594] Benzyl N-[[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)- To a solution of [3-hydroxy-cyclohexoxy]-5-azido-tetrahydropyran-2-yl]methyl]-N-benzyl-carbamate (180 mg, 0.160 mmol) and (2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoic acid (F-HABA, Example 5, 77.7 mg, 0.274 mmol) in anhydrous THF (12 mL) was added DMTMM chloride (77.7 mg, 0.281 mmol). The mixture was stirred at rt for 18 h and then diluted with saturated aqueous NH4Cl (5 mL) and DCM (30 mL). The separated aqueous layer was extracted with DCM (3 × 10 mL). The combined organic layers were washed with brine (5 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse phase column using a gradient of 10-100% MeCN in 10 mM ammonium formate, pH 3.8, and lyophilized to give the title compound (187 mg, 84%) as a white solid. m / z (ESI) [M+H] + :1390.6, HPLC A(05)tR=2.85min.

[0595] Step K: Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (compound 1).

[0596] [ka]

[0597] Benzyl N-[[(2S,5R,6R)-5-azido-6-[(1R,2R,3S,4S,5R,6S)-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-4-[[(2R,3R)-4-azido-3-fluoro-2-[( A solution of [4-methoxyphenyl]methoxy]butanoyl]amino]-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy-cyclohexoxy]tetrahydropyran-2-yl]methyl]-N-benzylcarbamate (187 mg, 134 μmol) in MeOH (6 mL) was added to a suspension of Pd(OH) (20% aqueous, 652 mg, 0.93 mmol) in a mixture of AcOH (4.8 mL) and water (2 mL). The mixture was degassed by 3× vacuum-H2 cycles and then bubbled with hydrogen for 10 min. The mixture was stirred under a hydrogen atmosphere (1 atm) for 3 h and then degassed with N2 for 5 min. The suspension was filtered onto a 0.40 μM PVDF filter, which was washed with MeOH (5 × 8 mL) and water (5 × 10 mL), and the volatiles were evaporated under reduced pressure (rotary evaporator bath temperature <35 °C). The material was purified by preparative HPLC using a gradient of 5-15% MeCN in 10 mM NH4HCO3, pH 10, and lyophilized to give a solid (100 mg). H2SO4 (0.05 M in HO) was added dropwise to a solution of the above solid in HO (3 mL) at 0 °C until pH 4, then lyophilized. The solid was dissolved in water (2 mL) and MeOH (10 mL) was added to give a precipitate. This precipitate was filtered through a PVDF syringe filter and washed with MeOH (2 × 5 mL). The mother liquor was discarded, and the solid was dissolved in water (5 mL), and the solution was lyophilized to give the title product, Compound 1 (67.2 mg, 49%), as an off-white solid. 1H NMR (400 MHz, D2O) δ 5.77 (d, J = 3.5 Hz, 1H), 5.24 (d, J = 1.9 Hz, 1H), 5.22 - 5.02 (m, 2H), 4.47 - 4.35 (m, 1H), 4.33 - 4.24 (m, 2H), 4.21 - 4.15 (m, 1H), 4.14 - 3.94 (m, 4H), 3.89 - 3.74 (m, 3H), 3.72 - 3.55 (m, 3H), 3.55 - 3.47 (m, 1H), 3.47 - 3.04 (m, 8H), 2.91 (dd, J = 13.4, 8.2 Hz, 1H), 2.01 - 1.69 (m, 3H), 1.44 (t, J = 12.6 Hz, 1H). Note: No exchangeable protons were observed.

[0598] Example 7. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 2).

[0599] [ka]

[0600] Step A: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0601] [ka]

[0602] (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-amino-6-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetra To a solution of (hydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (Example 6, Step I, 220 mg, 0.196 mmol), (2S)-2-benzyloxy-4-(benzyloxycarbonylamino)butanoic acid (HABA, 101 mg, 0.293 mmol), and DIPEA (0.10 mL, 0.587 mmol) in anhydrous DMF (4 mL) was added PyBOP (122 mg, 0.235 mmol) at 0° C. The mixture was stirred at rt for 1.5 h and then poured into water (5 mL) and EtOAc (5 mL). The separated aqueous layer was extracted with EtOAc (3×5 mL). The combined organic layers were washed with brine (5 mL), dried (MgSO), filtered, and concentrated under reduced pressure. The material was purified by reverse phase column (ACCQ) using ACN and ammonium formate (10 mM, pH 3.8) and lyophilized to give the title compound (142 mg, 50%) as a white solid. M+H+: 1451.7

[0603] Step B: Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (compound 2).

[0604] [ka]

[0605] (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino) A solution of (butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (142 mg, 0.10 mmol) in MeOH (6.0 mL) was added to a suspension of Pd(OH) (474 ​​mg, 0.67 mmol, 20% aqueous) in a mixture of AcOH (5.0 mL) and water (2.0 mL). Hydrogen was bubbled through the mixture for 10 min. The mixture was stirred under a hydrogen atmosphere (1 atm) for 3 h and then degassed with N for 5 min. The suspension was filtered onto a 0.22 μM PVDF filter syringe, which was washed with MeOH (3 × 3 mL) and water (2 × 2 mL). Volatiles were evaporated under reduced pressure (temperature < 35 °C). The material was purified by filtration using 5% MeCN in 10 mM NH4HCO3 (pH 10) and lyophilized to give a solid. H2SO4 (0.05 M in HO) was added dropwise to a solution of the above solid in HO (3 mL) at 0 °C until the pH reached 4-5, then lyophilized. The solid was dissolved in water (2 mL), and then MeOH (10 mL) was added to give a precipitate. The solid was filtered through a 0.22 μM PVDF syringe filter, rinsed with MeOH (2 × 5 mL), and then washed with MeOH (5 mL). The solid was dissolved in water (10 mL) and lyophilized to give the title compound 2 (58 mg, 60%) as a white solid. 1H NMR (400 MHz, D2O) δ 5.96 (d, J = 3.6 Hz, 1H), 5.44 (s, 1H), 5.33 (s, 1H), 4.62 - 4.54 (m, 1H), 4.50 - 4.45 (m, 1H), 4.45 - 4.35 (m, 2H), 4.30 - 4.12 (m, 4H), 4.03 - 3.90 (m, 3H), 3.88 - 3.76 (m, 3H), 3.69 (t, J = 9.8 Hz, 1H), 3.65 - 3.55 (m, 2H), 3.51 - 3.40 (m, 2H), 3.39 - 3.25 (m, 2H), 3.25 - 3.16 (m, 2H), 3.11 (dd, J = 13.5, 8.2 Hz, 1H), 2.30 - 2.17 (m, 1H), 2.14 - 2.02 (m, 3H), 1.97 (d, J = 14.7 Hz, 1H), 1.68 - 1.58 (m, 1H).

[0606] Example 8. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (Compound 3).

[0607] [ka]

[0608] Step A: Synthesis of (3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl(E)-2,2,2-trifluoro-N-phenylacetimidate

[0609] [ka]

[0610] Cesium carbonate (23.5 g, 72.2 mmol) was added to a solution of 2,2,2-trifluoro-N-phenyl-acetimidoyl chloride (10.4 mL, 64.9 mmol) and benzyl N-[(1S)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (Ring A2, Example 2, Step G, 14.8 g, 36.1 mmol) in DCM (336 mL) at 23 °C under nitrogen. The mixture was stirred at 23 °C for 45 min, filtered through a pad of diatomaceous earth, rinsed with DCM (200 mL), and concentrated. The residue was purified by silica gel chromatography (330 g high-capacity cartridge) using hexanes and EtOAc (0-20%) to give the title compound as a solid (21 g, quant.). m / z: no ES+ ionization; LCMS (A05) tR = 3.14 min.

[0611] Step B: Synthesis of (1S,2S,3R,4S,5S,6R)-6-(((3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-3,5-bis(((benzyloxy)carbonyl)amino)cyclohexane-1,2,4-triyl triacetate

[0612] [ka]

[0613] [(3R,6S)-3-Azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl](1E)-2,2,2-trifluoro-N-phenyl-ethaneimidate 2 (21.5 g, 37 mmol) and [(1S,2R,3S,4S,5R,6R)-3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (Ring B1, 31.7 g, 55.5 mmol) were coevaporated with anhydrous toluene (2 × 45 mL). The mixture was dried under reduced pressure for 30 min. The residue was taken up in anhydrous DCM (550 mL) and EtO (250 mL) and then transferred via syringe to a RBF containing pre-activated 4A sieves. The mixture was stirred at rt for 30 min, then Bi(OTf) (26.7 g, 40.7 mmol) was added in one portion at 0 °C. The mixture was stirred at rt for 3 h and then diluted with saturated aq NH Cl solution (500 mL) and DCM (400 mL). The suspension was filtered through a pad of diatomaceous earth and the layers were partitioned. The separated aqueous layer was extracted with DCM (3 × 300 mL). The combined organic layers were washed with brine (500 mL), dried (MgSO), filtered, and concentrated under reduced pressure to give crude 3 (56 g, crude) as a β / α = 1:9 anomeric mixture. The material was used in the next step without further purification. m / z: ES+ [M+H] + 965.3; LCMS (A05); tR = 3.00 min (β anomer), 3.06 min (α anomer).

[0614] Step C: Synthesis of benzyl N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy-cyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate

[0615] [ka]

[0616] Sodium methoxide (4.60 M in MeOH, 10 mL, 46.4 mmol) was added to a solution of [(1S,2S,3R,4S,5S,6R)-2-acetoxy-3-[(3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy-4,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (56 g, 58 mmol, crude) in MeOH (375 mL) at room temperature. The mixture was stirred at rt for 1 h, diluted with AcOH (10 mL) to pH 5, and then concentrated under reduced pressure. The residue was purified by preparative HPLC (X-bridge C18, 50 x 250 mm, 10 μM) using 58-72% ACN in ammonium formate (10 mM, pH 3.8). Fractions were pooled and concentrated under reduced pressure to remove the ACN. The aqueous residue was extracted with EtOAc (3 x 200 mL). The combined organic phases were washed with brine (2 x 100 mL), dried (MgSO), filtered, and concentrated under reduced pressure to give the title compound (17.5 g, 59%, over three steps) as a solid. m / z (ESI) [M+H] + ;839.3, HPLC A(05)tR=2.80min.

[0617] Step D: Synthesis of benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-5,7-bis(benzyloxycarbonylamino)-6-hydroxy-spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1'-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate

[0618] [ka]

[0619] To a solution of benzyl N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxycyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzylcarbamate (17.5 g, 20.9 mmol) and 1,1-dimethoxycyclohexane (26.9 mL, 177 mmol) in DME (337 mL) was added dry PPTS (2.17 g, 8.62 mmol). The mixture was stirred at 90 °C under nitrogen for 3 h. The mixture was cooled to rt and concentrated under reduced pressure. The residue was purified on silica gel (350 g, 25 μM HCl) using EtOAc and hexanes (0-100%) to give the title compound (17.5 g, 91%) as a white solid. m / z: ES+, [M+H] + 919.4; LCMS(A05); tR=3.12 min.

[0620] Step E: Synthesis of benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-6-benzyloxy-5,7-bis(benzyloxycarbonylamino)spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1'-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate

[0621] [ka]

[0622] Sodium hydride (60% dispersion in mineral oil, 1.85 g, 46.3 mmol) was added to a mixture of benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-5,7-bis(benzyloxycarbonylamino)-6-hydroxy-spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1′-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (18.5 g, 20.1 mmol) and benzyl bromide (26.8 mL, 226 mmol) in THF (500 mL) at 0° C. under nitrogen. The mixture was stirred at rt for 6 h (<50% conversion), transferred to a 30° C. oil bath, and stirred for 5 h (complete conversion), then AcOH (2.27 mL, 39.7 mmol) was added, followed by silica gel and EtOAc (150 mL). The mixture was concentrated and purified by silica gel chromatography (350 g cartridge, 25 μM) using a gradient of 0-100% EtOAc in hexanes. Mixed fractions were pooled, concentrated, and repurified by silica gel chromatography (350 g cartridge) using a gradient of 0-50% EtOAc in hexanes to give the title compound (16.8 g, 83%) as a solid. m / z: ES+ [M+H] + 1009.4;LCMS(A05);tR=3.23min.

[0623] Step F: Synthesis of dibenzyl((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate

[0624] [ka]

[0625] To a solution of benzyl N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxycyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzylcarbamate (17.5 g, 20.9 mmol) and 1,1-dimethoxycyclohexane (26.9 mL, 177 mmol) in DME (337 mL) was added dry PPTS (2.17 g, 8.62 mmol). The mixture was stirred at 90 °C under nitrogen for 3 h. The mixture was cooled to rt and concentrated under reduced pressure. The residue was purified on silica gel (350 g, 25 μM HCl) using EtOAc and hexanes (0-100%) to give the title compound (17.5 g, 91%) as a white solid. m / z: ES+, [M+H] + 919.4; LCMS(A05); tR=3.12 min.

[0626] Step G: Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate

[0627] [ka]

[0628] A solution of benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-6-benzyloxy-5,7-bis(benzyloxycarbonylamino)spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1′-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzylcarbamate (16.8 g, 16.6 mmol) in THF (180 mL), water (60 mL), and AcOH (150 mL) was heated at 80° C. for 12 h and then cooled to rt. The mixture was slowly added to water (2 L). The resulting precipitate was filtered, and the solid was washed with water (3×100 mL), collected, and then dissolved in EtOAc (500 mL). The solution was washed with saturated aq. NaHCO3 (100 mL) and water (3 x 100 mL). The aqueous layer was extracted with EtOAc (3 x 50 mL). The combined organic phases were washed with brine (2 x 50 mL), dried (MgSO4), filtered, and concentrated under reduced pressure to give the title product (14.8 g, 96%) as a solid, which was used directly without further purification. m / z (ESI) [2M+NH4] + :1875.3, HPLC(A05)tR=2.99min.

[0629] Step H: Synthesis of benzyl ((S)-1-((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (Ring AB2)

[0630] [ka]

[0631] Sodium hydride (60% oil dispersion, 3.19 g, 79.7 mmol) was added to a solution of benzyl N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4S,5S,6R)-3-benzyloxy-2,4-bis(benzyloxycarbonylamino)-5,6-dihydroxy-cyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (14.8 g, 15.9 mmol) in dry DMF (150 mL) at 0° C. The mixture was stirred at 0° C. for 1 h 30, diluted with AcOH (7.8 mL, 135 mmol), and then slowly poured into stirring water (1 L). The precipitate was filtered, and the solid was washed with water (3 x 200 mL) and lyophilized to give the title product (13.8 g, >99%) as a solid. The solid was further purified on a silica gel column using a gradient of 0-100% EtOAc in hexanes to give the title product (11 g, 84%) as a solid. m / z (ESI) [M+H] + :821.3, HPLC(A05)tR=2.90min.

[0632] Step I: Synthesis of [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy-4-benzyloxy-5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-7-yl]oxy]-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-tetrahydrofuran-2-yl]methyl acetate

[0633] [ka]

[0634] activation KCO (2.73 g, 19.7 mmol) was added to a mixture of [(2R,3R,4R)-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-5-hydroxy-tetrahydrofuran-2-yl]methyl acetate ring CD-1 (6.9 g, 13 mmol) and CClCN (2 mL, 19.7 mmol) in dry DCM (300 mL). The mixture was stirred at rt for 18 h, then filtered over Celite, rinsed with DCM, and concentrated under reduced pressure to give a syrup.

[0635] Glycosylation A mixture of this syrup and benzyl N-[(1S)-1-[(2S,5R,6R)-6-[[(3aR,4S,5S,6R,7S,7aS)-4-benzyloxy-5-(benzyloxycarbonylamino)-7-hydroxy-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-6-yl]oxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (2.7 g, 3.29 mmol) was coevaporated with dry toluene (12 mL) under reduced pressure, dissolved in dry DCM (300 mL), and then transferred to a RBF containing pre-activated 4A sieves. The mixture was cooled to −78 °C, and then BF₃·OEt₂ (6.12 mL, 49.6 mmol) was added slowly over 5 min at −78 °C. Upon completion of the addition, the dry ice bath was immediately removed. The mixture was stirred at rt for 2 h and diluted with a saturated aqueous solution of NaHCO₃ (150 mL) and DCM (300 mL). The mixture was filtered through Celite, and the cake was washed with DCM (3 × 150 mL). The mixture was partitioned, and the aqueous layer was extracted with DCM (3 × 50 mL). The combined organic layers were washed with brine (2 × 100 mL), dried (MgSO₄), filtered, and concentrated under reduced pressure. The material was purified by preparative HPLC (X-bridge C18, 50 × 250 mm, 10 μM) using 72–86% ACN in ammonium formate (10 mM, pH 3.8). The product fractions were pooled and the ACN was removed under reduced pressure. The aqueous layer was extracted with EtOAc (3×150 mL).The combined organic layers were washed with brine (2 x 50 mL), dried (MgSO4), filtered, and concentrated under reduced pressure to give the title compound, [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy-4-benzyloxy -5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-7-yl]oxy]-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-tetrahydrofuran-2-yl]methyl acetate (4 g, 91%). m / z (ESI) [M+H]. + :1334.2, HPLC(A05)tR=3.04min.

[0636] Step J: Synthesis of benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy-cyclohexoxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate

[0637] [ka]

[0638] [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy-4-benzyloxy-5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzo[ To a solution of [(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-tetrahydrofuran-2-yl]methyl acetate (4.0 g, 3.00 mmol) in dioxane (100 mL) and water (100 mL) was added LiOH·HO (3.78 g, 90.0 mmol). The mixture was stirred at 40 °C for 18 h, cooled to rt, diluted with EtOAc (300 mL) and HO (50 mL), and then partitioned. The separated aqueous layer was extracted with EtOAc (3 × 100 mL). The combined organic layers were dried (MgSO), filtered, and concentrated under reduced pressure to give the title compound (3.40 g, 99%) as a white solid. m / z(ESI)[M+H] + :1139.4, HPLC A(05)tR=2.54min.

[0639] Step K: (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanamide)-6-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl)oxy) Synthesis of di((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0640] [ka]

[0641] Benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxy) To a solution of (2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoic acid (1.37 g, 4.82 mmol) in anhydrous THF (150 mL) was added DMTMM chloride (1.37 g, 4.93 mmol). The mixture was stirred at rt for 18 h and then diluted with saturated aqueous NH4Cl (100 mL) and EtOAc (300 mL). The separated aqueous layer was extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (2 × 50 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse-phase preparative HPLC (X-bridge C18, 50 x 250 mm, 10 μM) using a gradient of 70-85% ACN in ammonium formate (10 mM, pH 3.8). Product fractions were pooled and concentrated under reduced pressure to remove ACN. The aqueous layer was extracted into EtOAc (3 x 150 mL). The combined organic phases were washed with brine (2 x 50 mL), dried (MgSO), filtered, and concentrated under reduced pressure to give the title compound (2.7 g, 68%) as a solid. m / z (ESI) [M+H] + :1404.4, HPLC A(05)tR=2.96min.

[0642] Step L: Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (compound 3)

[0643] [ka]

[0644] A mixture of MeOH (152 mL), AcOH (127 mL), and water (51 mL) was added via syringe under N2 to benzyl N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2R,3S,4S,5R,6S)-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl) The mixture was transferred to a RBF containing 2.8 g (1.99 mmol) of [(2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoyl]amino]-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxycyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzylcarbamate (2.8 g, 1.99 mmol) and Pd(OH) (20% aqueous, 8.4 g, 12 mmol). The mixture was degassed using three vacuum-H cycles, then bubbled with hydrogen (1 atm) for 4 h and then degassed with N for 15 min. The suspension was filtered onto Celite, which was washed with MeOH (200 mL) and water (200 mL). The volatiles were evaporated under reduced pressure (rotary evaporator bath temperature <35 °C). The material was purified by preparative HPLC using 5-15% MeCN in 10 mM ammonium bicarbonate (pH 10) and then lyophilized to give a solid. The solid was dissolved in water (50 mL), and then ammonium sulfate (3 eq, 790 mg, 5.98 mmol) was added. The mixture was stirred at rt for 5 min, then filtered through a 0.40 μM syringe filter and added dropwise to MeOH (1.5 L) with vigorous stirring. The suspension was filtered over a fine frit and rinsed with MeOH (50 mL). The mother liquor was discarded, and the solid was dissolved in water (40 mL) and lyophilized to give the title compound (1100 mg, 54%) as a solid. Compound 3 (1.1 g, 53%) as a white solid. 1H NMR (400 MHz, D2O) δ 5.87 (d, J = 3.6 Hz, 1H), 5.29 (s, 1H), 5.25 - 5.05 (m, 2H), 4.47 - 4.17 (m, 4H), 4.18 - 3.97 (m, 3H), 3.88 - 3.78 (m, 3H), 3.77 - 3.68 (m, 3H), 3.62 (dd, J = 12.5, 5.6 Hz, 1H), 3.54 (t, J = 9.9 Hz, 1H), 3.49 - 3.44 (m, 1H), 3.44 - 3.16 (m, 7H), 1.99 - 1.82 (m, 3H), 1.49 - 1.34 (m, 1H), 1.19 (d, J = 6.7 Hz, 3H). Note: 20 exchangeable protons were not observed.

[0645] Example 9. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (Compound 4).

[0646] [ka]

[0647] Step A: Synthesis of (3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl(E)-2,2,2-trifluoro-N-phenylacetimidate

[0648] [ka]

[0649] To a mixture of benzyl N-[(1R)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (Ring A3, Example 3, 1.30 g, 2.85 mmol) and CsCO (1.86 g, 5.70 mmol) in DCM (10.0 mL) was added 2,2,2-trifluoro-N-phenylacetimidoyl chloride (1.19 g, 5.70 mmol). The mixture was stirred at rt for 2 h. The mixture was filtered, the solids were rinsed with DCM, and the solvent was concentrated under reduced pressure. The residue was purified on silica gel (dry loading) using a gradient of 0-35% EtOAc in hexanes to give the title compound (1.20 g, 73%) as an oil. MS: No ionization.

[0650] Step B: Synthesis of dibenzyl ((1R,2R,3S,4R,5R,6S)-4-(((3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,5,6-trihydroxycyclohexane-1,3-diyl)dicarbamate

[0651] [ka]

[0652] [[[(2R,3R,6S)-3-Azido-6-[(1R)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl](1E)-2,2,2-trifluoro-N-phenyl-ethaneimidate (2.47 g, 4.25 mmol) and [(1S,2R,3S,4S,5R,6R)-3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (Ring B1, 3.28 g, 5.73 mmol) were coevaporated with anhydrous toluene (2 × 3 mL). The mixture was dried under reduced pressure for 30 min. The residue was taken up in anhydrous DCM (100 mL) and then transferred via syringe to a RBF containing pre-activated 3 Å and 4 Å sieves. EtO (60 mL) was added. The mixture was stirred at rt for 20 min and then cooled to 0 °C. Bi(OTf) (3.07 g, 4.67 mmol) was added in one portion, and the mixture was stirred at rt for 4 h. A saturated aqueous solution of NH Cl (100 mL) was added. The mixture was filtered over diatomaceous earth. The separated aqueous layer was extracted with DCM. The combined organic layers were dried (MgSO), filtered, and concentrated under reduced pressure. The residue was purified on silica gel using a gradient of 0 to 100% EtOAc in hexanes. The solid was dissolved in MeOH (100 mL), and K CO was added (1.52 g, 11.0 mmol). The mixture was stirred at rt for 2 h. The mixture was concentrated under reduced pressure (the rotary evaporator water bath did not exceed 35 °C). Water (100 mL) and EtOAc (100 mL) were added. The separated aqueous layer was extracted with EtOAc. The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure to give the title compound (2.30 g, 99%) as a white solid. m / z (ESI) [M+H] + ;839.3.

[0653] Step C: Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1′-cyclohexane]-5,7-diyl)dicarbamate

[0654] [ka]

[0655] A mixture of benzyl N-[(1R)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxycyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzylcarbamate (2.30 g, 2.60 mmol), 1,1-dimethoxycyclohexane (2.38 mL, 15.6 mmol), and PPTS (262 mg, 1.01 mmol, dried over V10) in dry DME (111 mL) was stirred at 90 °C for 30 min. The mixture was cooled to rt and concentrated under reduced pressure. The material was purified by ACCQ using 10 mM ammonium formate, pH 3.8, and MeCN. The product was then diluted in EtOAc and washed with brine. The organic layer was dried (MgSO4), filtered and concentrated under reduced pressure to give (1.40 g, 59%). m / z (ESI) [M+H] + ;919.4.

[0656] Step D: Synthesis of dibenzyl((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxole-2,1′-cyclohexane]-5,7-diyl)dicarbamate

[0657] [ka]

[0658] Sodium hydride (60% dispersion in mineral oil, 141 mg, 3.51 mmol) was added to a mixture of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1′-cyclohexane]-5,7-diyl)dicarbamate (1.40 g, 1.52 mmol) and BnBr (2.03 mL, 17.1 mmol) in THF (75 mL) at 0° C. under nitrogen. The mixture was stirred at 0° C. for 4 h, and then AcOH (0.24 mL, 4.13 mL) was added. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (120 g cartridge) using a gradient of 0-60% EtOAc in hexanes to afford the title compound (1.43 g, 93%) as a solid. m / z (ESI) [M+H] + ;1009.5.

[0659] Step E: Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate

[0660] [ka]

[0661] A mixture of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxole-2,1′-cyclohexane]-5,7-diyl)dicarbamate (1.44 g, 1.43 mmol) in AcOH (18 mL), water (8.0 mL), and THF (27 mL) was heated at 80° C. for 24 h. The mixture was cooled to rt, and the THF was removed under reduced pressure. Water (100 mL) was added, and a suspension was observed. The suspension was filtered over a medium frit. The solid was rinsed with water and dried under vacuum to give the title compound (1.21 g, 91%). m / z: ES+ [M+H] + 929.6.

[0662] Step F: Synthesis of benzyl ((R)-1-((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (Ring AB3)

[0663] [ka]

[0664] NaH (210 mg, 5.25 mmol, 60% oil dispersion) was added to a solution of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate (1.22 g, 1.31 mmol) in dry DMF (20 mL) at 0° C. The mixture was stirred at 0° C. for 3 h, and then AcOH (0.64 mL, 11.2 mmol) was added. The solution was poured into water (200 mL). The suspension was filtered, the solid rinsed with water (20 mL) and dried under reduced pressure to give the title compound (830 mg, 77%) as a white solid. m / z (ESI) [M+H] + :821.3.

[0665] Step G: Synthesis of benzyl ((1R)-1-((2S,5R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate

[0666] [ka]

[0667] To a mixture of [(2R,3R,4R)-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-5-hydroxy-tetrahydrofuran-2-yl]methyl acetate (Ring CD1, WO 2019194858, incorporated by reference, 961 mg, 1.81 mmol) and KCO (429 mg, 3.11 mmol) in dry DCM (8 mL) was added CClCN (0.31 mL, 3.11 mmol). The mixture was stirred at rt for 18 h, then filtered over diatomaceous earth, rinsed with DCM, and concentrated under reduced pressure. Benzyl ((R)-1-((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (Ring AB3, 425 mg, 0.518 mmol) in dry DCM (8 mL) was added to the above crude material and then transferred to an RBF containing pre-activated 3A and 4A sieves. The mixture was cooled to −78 °C, and then BF-OEt (0.96 mL, 7.81 mmol) was added dropwise at −78 °C. Upon completion of the addition, the dry ice bath was immediately removed. The mixture was stirred at rt for 1 h, diluted with a saturated aqueous solution of NaHCO3 (10 mL), filtered through diatomaceous earth, and the filter cake was washed with DCM (20 mL). The separated aqueous layer was extracted with DCM (3 × 5 mL). The combined organic layers were washed with brine (15 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The residue was taken up in 1,4-dioxane (4.0 mL) and water (4.0 mL), then LiOH-H2O (652 mg, 15.5 mmol) was added, and the mixture was stirred at 40 °C for 18 h. The mixture was cooled to rt, then the 1,4-dioxane was removed under reduced pressure, and then EtOAc (10 mL) was added. The separated aqueous layer was extracted with EtOAc (3 × 10 mL). The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure.The residue was purified by ACCQ prep using a gradient of MeCN in 10 mM ammonium bicarbonate, pH 10 to give the title compound (209 mg, 35% over two steps) as a solid. m / z (ESI) [M+H]. + :1139.5

[0668] Step H: 2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanamide)-6-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl Synthesis of tetrahydro-2H-pyran-3,4-diyl diacetate (4-benzyloxy)-5-((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0669] [ka]

[0670] To a solution of benzyl ((1R)-1-((2S,5R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (104 mg, 0.091 mmol) and (2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoic acid (F-HABA, Example 5, 44 mg, 0.157 mmol) in anhydrous THF (5 mL) was added DMTMM chloride (44 mg, 0.160 mmol). The mixture was stirred at rt for 18 h and then diluted with saturated aqueous NH4Cl (3 mL) and EtOAc (5 mL). The separated aqueous layer was extracted with EtOAc (3 x 4 mL). The combined organic layers were washed with brine (5 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by preparative HPLC (ACCQ, BEH C18 column) using MeCN in ammonium formate (10 mM, pH 3.8) and lyophilized to afford the title compound (103 mg, 80%) as a white solid. m / z (ESI) [M+H] + :M+H+:1404.6.

[0671] Step I: Synthesis of ((2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (compound 4)

[0672] [ka]

[0673] 2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanamido)-6-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyl A solution of (oxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (104 mg, 0.074 mmol) in MeOH (9.0 mL) was added to a suspension of Pd(OH) (359 mg, 0.51 mmol, 20% aqueous) in a mixture of AcOH (7.50 mL) and water (3.0 mL), degassed by 3× vacuum-H cycles, and then bubbled with hydrogen for 10 min. The mixture was stirred under a hydrogen atmosphere (1 atm) for 2.5 h, degassed with N for 5 min, and then filtered onto a 0.22 μM PVDF filter syringe, which was washed with MeOH (3 × 3 mL) and water (2 × 2 mL), and the volatiles were evaporated under reduced pressure (rotary evaporator bath temperature <35 °C). The material was purified by filtration using 5% MeCN in 10 mM NHHCO (pH 10) and lyophilized to give a solid. HSO (0.05 M in HO) was added dropwise to a solution of the above solid in HO (3 mL) at 0 °C until the pH reached 4, and then lyophilized to give a solid, which was dissolved in water (2 mL) followed by MeOH (10 mL) to give a precipitate. The precipitate was filtered through a 0.22 μM PVDF syringe filter, rinsed with MeOH (2 × 5 mL), and then washed with water (5 mL). The solid was dissolved in water (10 mL) and lyophilized to give the title compound 4 (30 mg, 40%) as a white solid. 1H NMR (500 MHz, D2O) δ 5.95 - 5.88 (m, 1H), 5.47 - 5.24 (m, 3H), 4.57 (d, J = 10.2 Hz, 1H), 4.51 - 4.34 (m, 3H), 4.27 (d, J = 3.7 Hz, 2H), 4.18 (d, J = 12.1 Hz, 1H), 4.04 - 3.88 (m, 5H), 3.82 - 3.59 (m, 5H), 3.58 - 3.37 (m, 5H), 3.33 - 3.22 (m, 2H), 2.12-2.00 (m, 2H), 1.96 - 1.89 (m, 1H), 1.69 - 1.58 (m, 1H), 1.31 (d, J = 6.9 Hz, 3H).

[0674] Example 10 Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 5)

[0675] [ka]

[0676] Step A: (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4 Synthesis of -(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0677] [ka]

[0678] Benzyl N-[(1R)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)- To a solution of [3-hydroxy-cyclohexoxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (90 mg, 0.079 mmol), (2S)-2-benzyloxy-4-(benzyloxycarbonylamino)butanoic acid (HABA, 40.7 mg, 0.119 mmol), and DIPEA (0.04 mL, 0.237 mmol) in anhydrous DMF (3 mL) was added PyBOP (49.3 mg, 0.0948 mol) at 0 °C. The mixture was warmed to rt and stirred for 18 h. The mixture was poured into water (5 mL). The pH was adjusted to 4 using 1 N HCl, and then EtOAc (40 mL) was added. The separated aqueous layer was extracted with EtOAc (5 mL × 3). The combined organic layers were washed with brine (5 mL), dried (MgSO), filtered, and concentrated under reduced pressure. The material was purified by reverse-phase column (ACCQ) using a gradient of 60-80% ACN in ammonium formate (10 mM, pH 3.8) and concentrated under reduced pressure. The material was then dissolved in EtOAc (30 mL) and water (5 mL). The separated aqueous layer was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried (MgSO), filtered, and concentrated under reduced pressure to give the title compound (79.3 mg, 69%) as a white solid. m / z (ESI) [M+H] + :1464.5, HPLC A(05)tR=2.65min.

[0679] Step B: Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 5)

[0680] [ka]

[0681] (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butanamide)-3-(((benzyl A solution of (6-hydroxycyclohexyl)-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (79 mg, 0.0539 mmol) in MeOH (6.5 mL) was added to a suspension of Pd(OH) (20% aqueous, 261 mg, 0.372 mmol) in a mixture of AcOH (5.5 mL) and water (2.2 mL), degassed by 3× vacuum-H cycles, and then bubbled with hydrogen for 10 min. The mixture was stirred for 4 h while bubbling with hydrogen (1 atm, refilling the balloon with H as needed). The suspension was degassed with N2 for 15 min and then filtered onto a 0.22 μM PVDF filter syringe, which was washed with MeOH (3 × 5 mL) and water (2 × 5 mL), and the volatiles were evaporated under reduced pressure (rotary evaporator bath temperature <35 °C). The material was purified by filtration using 5% MeCN in 10 mM NH4HCO3 (pH 10) and lyophilized to give a solid. H2SO4 (0.05 M in HO) was added dropwise to a solution of the above solid in HO (3 mL) at 0 °C until pH 4 was reached, followed by lyophilization. The solid was dissolved in water (2 mL) followed by MeOH (10 mL). The suspension was filtered through a 0.22 μM PVDF syringe filter, rinsed with MeOH (2 × 5 mL), and then washed with water (5 mL). The solid was dissolved in water (10 mL) and lyophilized to give the title product, Compound 5 (26 mg, 48%) as a solid. 1H NMR (400 MHz, D2O) δ 5.76 (dd, J = 10.0, 3.7 Hz, 1H), 5.27 (dd, J = 6.2, 2.8 Hz, 1H), 5.16 (d, J = 1.8 Hz, 1H), 4.40 (dd, J = 7.2, 4.7 Hz, 1H), 4.35 - 4.28 (m, 1H), 4.26 - 4.18 (m, 2H), 4.10 (t, J = 3.3 Hz, 2H), 4.03 - 3.95 (m, 1H), 3.93 - 3.67 (m, 5H), 3.64 - 3.19 (m, 8H), 3.18 - 3.08 (m, 1H), 3.08 - 2.98 (m, 2H), 2.13 - 1.99 (m, 1H), 1.98 - 1.84 (m, 3H), 1.81 - 1.70 (m, 1H),1.55 - 1.40 (m, 1H), 1.14 (dd, J = 6.9, 1.2 Hz, 3H). m / z (ESI) [M+H] + : 714.4.

[0682] Example 11 Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 6)

[0683] [ka]

[0684] Step A: (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4 Synthesis of -(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0685] [ka]

[0686] Benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy To a solution of [-cyclohexoxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (Example 8, Step J, 133 mg, 0.117 mmol), (2S)-2-benzyloxy-4-(benzyloxycarbonylamino)butanoic acid (HABA; 60.1 mg, 0.175 mmol), and DIPEA (0.061 mL, 0.350 mmol) in anhydrous DMF (4.0 mL) was added PyBOP (73.0 mg, 0.140 mol) at 0 °C. The mixture was warmed to rt and stirred for 18 h. The mixture was poured into water (5.0 mL), followed by the addition of EtOAc (5.0 mL). The separated aqueous layer was extracted with EtOAc (3 × 5.0 mL). The combined organic layers were washed with brine (5.0 mL), dried (MgSO), filtered, and concentrated under reduced pressure. The material was purified by reverse-phase column (C18, 40 g) using a gradient of 0-100% ACN in ammonium formate (10 mM, pH 3.8), concentrated under reduced pressure, and lyophilized to give the title compound (66 mg, 38%) as a white solid. m / z (ESI) [M+H] + :1464.5, HPLC A(05)tR=2.94min.

[0687] Step B: Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 6)

[0688] [ka]

[0689] (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)a A solution of (amino)butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (66 mg, 0.0451 mmol) in MeOH (7 mL) was added to a suspension of Pd(OH) (20% aqueous, 218 mg, 0.311 mmol) in a mixture of AcOH (5.0 mL) and water (3.0 mL). Hydrogen was bubbled through for 3 h, then degassed with N for 15 min. The suspension was filtered onto a 0.22 μM PVDF filter syringe, which was washed with MeOH (8.0 mL), water (8.0 mL), and DCM (8.0 mL), and then the volatiles were evaporated under reduced pressure (rotary evaporator, bath temperature less than 35° C.). The crude material was purified by filtration using 5% MeCN in 10 mM NH4HCO3 (pH 10) and lyophilized to give a solid. H2SO4 (0.05 M in HO) was added dropwise to a solution of the above solid in HO (3 mL) at 0° C. until pH 4, and then lyophilized. The solid was dissolved in water (2 mL), followed by MeOH (10 mL). The suspension was filtered through a 0.22 μM PVDF syringe filter, rinsed with MeOH (2 × 5 mL), and then washed with water (5 mL). The solid was dissolved in water (10 mL) and lyophilized to give the title product, Compound 6 (30 mg, 66%) as a solid. 1H NMR (500 MHz, D2O) δ 6.04 (d, J = 3 Hz, 1H), 5.47 (m, 1H), 5.33 (m, 2H), 4.58 - 4.55 (m, 1H), 4.48 - 4.34 (m, 3H), 4.30 - 4.21 (m, 3H), 4.05 - 3.85 (m, 6H), 3.81 - 3.66 (m, 2H), 3.63 (m, 1H), 3.61- 3.14 (m, 7H), 2.26 - 2.16 (m, 1H), 2.11 - 2.01 (m, 4H), 1.63 - 1.50 (m, 1H), 1.35 (d, J = 6.0 Hz, 3H). Note: 20 exchangeable protons were not observed. m / z (ESI) [M+H] + : 714.3, HPLC A(05) tR=0.33 min.

[0690] Example 12: Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((S)-1-(methylamino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (Compound 7)

[0691] [ka]

[0692] Step A: Synthesis of (S)-N-[(1S)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfinamide

[0693] [ka]

[0694] Sodium hydride (60% in mineral oil, 19.2 g, 500 mmol) was added to a mixture of (S)—N-[(1S)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (83.0 g, 334 mmol) in THF (1.0 L) at 0° C. under nitrogen. The mixture was stirred at 0° C. for 15 min. Iodomethane (41.5 mL, 667 mmol) was added, and the mixture was stirred at rt for 3 h. The mixture was diluted with saturated aqueous NH4Cl (500.0 mL) and water (200.0 mL), and the aqueous phase was extracted with ethyl acetate (3×250 mL). The combined organic phases were washed with brine (250 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The crude residue (94.3 g) was split into two portions, and each fraction was purified by silica gel chromatography (350 g, Biotage HC, liquid load) using a gradient of 0-30% EtOAc in hexanes to give the title compound. The fractions were combined. A third round of purification was performed on the combined fractions, which were purified by silica gel chromatography (350 g HC, liquid load) using a gradient of 0-20% EtOAc in hexanes. All pure fractions from the three purifications were combined to give the title compound (30.2 g, 31%, approximately 85% pure). m / z: ES+[M+H]+246.2; HPLC (A05); tR=2.29 min (desired title product). m / z: ES+[M+H]+246.2; HPLC (A05); tR=2.34 min (undesired diastereoisomer).

[0695] Step B: Synthesis of (S)-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfinamide

[0696] [ka]

[0697] Iodine (31.4 g, 124 mmol) was added portionwise to a suspension of (S)—N-[(1S)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfinamide (30.4 g, 124 mmol) and NaHCO (31.2 g, 372 mmol) in ACN (500.0 mL) and HO (500.0 mL) at 0° C. The mixture was stirred at 0° C. for 30 min. The mixture was stirred at rt for 2 h 30. The mixture was diluted with saturated aqueous NaSO (500.0 mL), and the aqueous layer was extracted with EtOAc (3 × 500.0 mL). The combined organic phase was washed with brine (300.0 mL), dried (MgSO4), filtered and concentrated to give the title compound as an oil (47.7 g, 99% crude). The compound was used without any further purification. m / z (ESI) [M+Na] + : 412.0, HPLC tR = 2.29 min and 2.40 min (desired product and diastereoisomer).

[0698] Step C: Synthesis of benzyl-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate

[0699] [ka]

[0700] 1 M aqueous HCl (147.0 mL, 147 mmol) was added to a solution of (S)-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfinamide (47.7 g, 123 mmol) in dioxane (750.0 mL) at 5–10°C. The mixture was stirred at 5–10°C for 45 min, and then PBS 10× buffer (approximately 2.0 L) was added until the pH reached approximately 6. The mixture was warmed to rt, and then CbzCl (35.0 mL, 245 mmol) was added. The mixture was stirred at rt for 3 h, and then EtOAc (500.0 mL) was added. The separated aqueous layer was extracted with EtOAc (3 × 400.0 mL). The combined organic layers were washed with brine, dried (MgSO), filtered, and concentrated under reduced pressure. The residue was split into two portions (2 x 45 g crude) and then purified on silica (330 g, liquid-loaded) using EtOAc and hexanes (0-50%) to give the title compound (15.0 g, 27%) as an oil. m / z (ESI) [M+Na]: 442.0, HPLC, A(05)tR = 2.49 m, 2.54 min. (title product).

[0701] Step D: Synthesis of benzyl-N-[(1S)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate

[0702] [ka]

[0703] Oxidation: To a mixture of benzyl-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate (15.0 g, 35.8 mmol) and 4 Å sieves (5.0 g) in DCM (250.0 mL) was added PDC (33.6 g, 89.4 mmol). The mixture was stirred at rt for 18 h, then filtered over a pad of silica gel, rinsed with EtOAc (3×350.0 mL), and concentrated under reduced pressure. m / z (ESI) [M+H] +: 418.0, HPLC (A05); tR = 2.44 min (oxidized intermediate).

[0704] Azide The residue was taken up in DMF (150.0 mL) and cooled to 0 °C. NaN (2.21 g, 34.0 mmol) was added, and the mixture was stirred for 1 h at 0 °C. Brine (500.0 mL) and EtO (150.0 mL) were added. The separated aqueous layer was extracted with EtO (2 × 150.0 mL). The combined organic layers were dried (MgSO) and concentrated under reduced pressure. The material was purified by MPLC on silica (220 g, Biotage HC) using a gradient of 0-60% EtOAc in hexanes to give the title compound (8.0 g, 67%, over two steps). m / z (ESI) [M+K] + :371.4, HPLC (A05); tR=2.33min (title product).

[0705] Step E: Synthesis of benzyl-N-[(1S)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate

[0706] [ka]

[0707] DIBAL-H (1 M in toluene, 43.3 mL, 43.3 mmol) was added dropwise over 20 min to a solution of benzyl-N-[(1S)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate (8.0 g, 24.1 mmol) in DCM (250.0 mL) at −78° C. After 1 h, EtOH (4.2 mL) was added dropwise at −78° C. The mixture was poured into a saturated aqueous solution of Rochelle's salt (350.0 mL, conc. 730 g / L in HO). The mixture was stirred for 18 h. The separated aqueous layer was extracted with DCM (3 × 150.0 mL). The combined organic layers were washed with brine, dried over MgSO, and concentrated under reduced pressure. The residue was purified by MPLC on silica gel (330 g, Biotage HC, dry loading) using 0-30% EtOAc in hexanes (gradient 50 min) to give the title compounds (two diastereoisomers) (3.3 g, 41% of the desired diastereoisomeric pair and 1.9 g, 24% of the undes...

Claims

1. Formula (III) 【Chemistry 1】 (In the formula, R ａ is H or methyl, R ｂ is H, C １～６ alkyl, or of formula (II) 【Chemistry 2】 The structure is Q is NH, O or S; n is an integer from 0 to 4, R １ is hydrogen or C １～３ is alkyl, R ２ and R ３ is independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R ４ is H, C １～３ Alkyl or -C(=NH)NR ４ａ R ４ｂ and R ４ａ and R ４ｂ is hydrogen and C １～３ alkyl, or R １ and R ４ together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N), or a pharma- ceutically acceptable salt or solvate thereof.

2. R ａ The compound of claim 1 , wherein

3. R ａ The compound of claim 1 , wherein is methyl.

4. The compound of formula (III) 【Chemistry 3】 The compound of any one of claims 1 to 3, having a structure selected from the group consisting of:

5. R ｂ The compound according to any one of claims 1 to 5, wherein is ethyl.

6. The compound of formula (III) 【Chemistry 4】 The compound of claim 5 having a structure selected from the group consisting of:

7. R ｂ but, 【Chemistry 5】 The compound according to any one of claims 1 to 3, selected from the group consisting of:

8. R ｂ but, 【Chemistry 6】 The compound according to any one of claims 1 to 3, selected from the group consisting of:

9. The compound of formula (III) 【Chemistry 7】 9. The compound of claim 8 having a structure selected from the group consisting of:

10. Formula (I) 【Chemistry 8】 (In the formula, R ａ is hydrogen (H) or methyl, R ｂ is C １～６ alkyl, or of formula (II) 【Chemistry 9】 The structure is Q is NH, O or S; n is an integer from 0 to 4, R １ is hydrogen or C １～３ is alkyl, R ２ and R ３ is independently selected from the group consisting of hydrogen, alkyl, halogen, and —OH; R ４ is H, C １～３ Alkyl or -C(=NH)NR ４ａ R ４ｂ and R ４ａ and R ４ｂ is H and C １～３ alkyl, or R １ and R ４ together with the atom to which they are attached form a heterocycloalkyl ring system containing at least one N), or a pharma- ceutically acceptable salt or solvate thereof.

11. R ａ The compound of claim 10, wherein is H.

12. R ａ The compound of claim 10, wherein is methyl.

13. R ｂ but, 【Chemistry 10】 The compound according to any one of claims 10 to 12, selected from the group consisting of:

14. R ｂ but, 【Chemistry 11】 The compound according to any one of claims 10 to 13, selected from the group consisting of:

15. The compound of formula (I) 【Chemistry 12】 11. The compound of claim 10 having a structure selected from the group consisting of:

16. R ｂ The compound according to any one of claims 10 to 12, wherein is ethyl.

17. The compound of formula (I) has the structure 【Chemistry 13】 17. The compound of claim 16 having the formula:

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 17 or a pharma- ceutically acceptable salt, solvate, tautomer or stereoisomer thereof and at least one pharma- ceutically acceptable excipient.

19. 19. A method for treating a bacterial infection in a subject in need thereof, comprising administering to the subject a compound according to any one of claims 1 to 17 or a pharma- ceutically acceptable salt, solvate, tautomer or stereoisomer thereof, or a pharmaceutical composition according to claim 18.

20. 20. The method of claim 19, wherein the bacterial infection is caused by a gram-positive bacterium, a gram-negative bacterium, an aerobic bacterium, or a facultative anaerobic bacterium.

21. The bacterial infection is selected from the group consisting of Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, 20. The method of claim 19, wherein the infection is caused by a species of the genus Salmonella, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Burkholderia or Enterococcus.

22. The bacterial infection may be caused by Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, Proteus mirabilis, Bacillus anthracis, P.

21. The method of claim 20, wherein the infection is caused by P. aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei, or Yersinia pestis.

23. 21. The method of claim 20, wherein the bacterial infection is a gram-negative bacterial infection.

24. 24. The method of claim 23, wherein the gram-negative bacterial infection is an infection with Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxidans.

25. The method according to any one of claims 19 to 24, wherein the bacterial infection is caused by a multidrug resistant (MDR) bacteria.

26. 23. The method of any one of claims 19 to 22, wherein the bacterial infection is an infection caused by extended-spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae.

27. 22. The method of claim 21, wherein the bacterial infection is caused by Staphylococcus aureus.

28. 28. The method of claim 27, wherein the Staphylococcus aureus is methicillin-resistant Staphylococcus aureus (MRSA).

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