WO2025046492A1 - Broad spectrum aminoglycosides and uses thereof - Google Patents

Abstract

Provided herein are aminoglycoside compounds, such as compounds of Formula I, Ia, Ib, Ic, and Id, and pharmaceutically acceptable salts thereof, useful as therapeutic and/or prophylactic agents. The compounds may be useful in treating a bacterial infection in a subject, for example a Gram-negative bacterial infection. Also provided herein are methods for preparation of such compounds.

Description

AttyDktNo. R1119431100WO BROAD SPECTRUM AMINOGLYCOSIDES AND USES THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to U.S. Provisional Application No. 63/535,809, filed August 31, 2023, and which is herein incorporated by reference in its entirety. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT [0002] 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. TECHNICAL FIELD [0003] The present disclosure relates generally to aminoglycoside compounds having antibacterial activity, and to uses thereof in treatment of medical conditions associated with pathogenic microorganisms. BACKGROUND [0004] The rapid spread of antibiotic resistance has prompted a continuing search for new agents capable of antibacterial activity. Aminoglycosides (AGs) are highly potent, broad-spectrum antibiotics with many desirable properties for the treatment of 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 the bacterial ribosome. For the structurally related antibiotics neamine, ribostamycin, neomycin B, and paromomycin, the binding site has been localized to the A-site of the prokaryotic 16S ribosomal decoding region RNA (see Moazed et al., Nature, 1987, 327, 389). Binding of AGs to this RNA target interferes with the fidelity of mRNA translation and results in miscoding and truncation, leading ultimately to bacterial cell death (see Alper et al. J. Am. Chem. Soc., 1998, 120, 1965). [0005] Over time, resistance to AGs has emerged in the clinic and degraded our antibacterial armamentarium. AG resistance (AG-R) can be categorized mechanistically into two primary types: (1) AG-modifying enzymes (AMEs), which modify and inactive the drug; and (2) enzymes called ribosomal methyltransferases (RMTs), which modify the drug target binding pocket. At present, the most common form of AG-R amongst clinical isolates is conferred by AMEs, which are diverse and widely distributed. RMTs, while less common than AMEs in some species, are a serious threat because they modify the 16S rRNA AG binding site and prevent AG target engagement, conferring high-level resistance to all clinically available parenteral AGs. As an additional concern, AME and RMT genes are typically located on plasmids or transposons together with genes encoding resistance to other classes of antibacterials, which leads to multidrug resistant isolates. [0006] Further, AGs can be toxic to the kidney (nephrotoxicity) and cochlea (ototoxicity). These are cumulative processes, with the likelihood of observing toxicity increasing with treatment duration. The AttyDktNo. R1119431100WO nephrotoxic potential of AGs limits the dose and the length of treatment, making it challenging to achieve the systemic exposures required for efficacy against some infections. Quite often, but not always, the structure- activity relationships that lead to increased antibacterial potency are correlated with those that cause toxicity to human kidney cells. The origin of this toxicity is assumed to result from a combination of different factors and mechanisms such as interactions with phospholipids, inhibition of phospholipases and the formation of free radicals. [0007] For the foregoing reasons, while progress has been made in this field, there is a need for new chemical entities that possess antibacterial activity against highly resistant organisms and an acceptable safety profile. SUMMARY [0008] The present disclosure relates to aminoglycoside compounds, including stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof, having antibacterial activity, and to uses thereof in, for example, treatment of medical conditions associated with a pathogenic microorganism, which are also referred to herein as "bacterial infections." [0009] In one aspect is provided a compound having a structure according to Formula I: , or a pharmaceutically

R¹ and R² are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, - C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R³ and R⁴ are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, - C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R⁵ is hydrogen or is C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, - OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and –CN; R⁶ is independently selected for each occurrence from the group consisting of hydrogen, C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl, wherein each of said C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl is optionally AttyDktNo. R1119431100WO substituted with one or more substituents independently selected for each occurrence from halogen, -OH, - CN, -NO₂, and -NH₂; R⁷ is selected from the group consisting of hydrogen, -OR⁶, and -N(R⁶)₂; R⁸ is selected from the group consisting of hydrogen and -OR⁶; R^a is a tetrahydrofuranyl ring or a hexahydropyranyl ring, connected via a glycosidic bond; R^b is a second tetrahydrofuranyl or hexahydropyranyl ring connected to R^a through a glycosidic bond; wherein each of R^a and R^b is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OR⁶, -N(R⁶)_2, and C_1-6 alkyl, wherein said C_1-6 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and R^c is hydrogen, C1-6 alkyl, a protecting group, or has a structure according to Formula II: , wherein:

Q is NH, O, or S; n is an integer from 0 to 4; R⁹ is hydrogen or C_1-3 alkyl; R¹⁰ and R¹¹ are each selected independently for each occurrence from the group consisting of hydrogen, alkyl, halogen, and –OH; and R¹² is H, C_1-3 alkyl, or –C(=NH)NR^12aR^12b, wherein R^12a and R^12b are each independently selected from the group consisting of hydrogen and C_1-3 alkyl; or R⁹ and R¹², together with the atoms to which they are attached, form a heterocycloalkyl ring system comprising at least one N. [0010] In some embodiments, R¹ and R² are both hydrogen. [0011] In some embodiments, R¹ and R² are both methyl. [0012] In some embodiments, one of R¹ and R² is hydrogen, and the other of R¹ and R² is methyl. [0013] In some embodiments, R³ is hydrogen and R⁴ is methyl. [0014] In some embodiments, R¹, R² and R³ are hydrogen, and R⁴ is methyl. [0015] In some embodiments, R¹ and R³ are both hydrogen and R² and R⁴ are both methyl. [0016] In some embodiments, R⁵ is hydrogen. [0017] In some embodiments, R^c is selected from the group consisting of: AttyDktNo. R1119431100WO . . to Formula Ia:

, wherein: 1

R ³ and R¹⁴ are ₆ said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of –OR⁶, N(R⁶)₂, and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, -OR⁶, -N(R⁶)₂, -NO₂, and –CN. [0020] In some embodiments, R¹³ is H or OH, and R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl. [0021] In some embodiments, R' and R'' are each hydrogen. [0022] In some embodiments, R' is hydrogen and R'' is methyl. [0023] In some embodiments, R' and R'' are each methyl. [0024] In some embodiments, R¹³ is -OH and R¹⁴ is -CH₂OH. [0025] In some embodiments, the compound of Formula I has a structure according to Formula Ib: AttyDktNo. R1119431100WO , wherein: R¹ is hydrogen or

R⁷ is NH2 or OH; R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of -OH, -NH₂, and C_1-3 alkyl, said C_1-3 alkyl optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OH, -NH₂, -NO₂, and CN. [0026] In some embodiments, R⁷ is NH₂. [0027] In some embodiments, R⁵ is H. [0028] In some embodiments, R⁵ is H and R⁷ is NH2. [0029] In some embodiments, the compound of Formula I has a structure according to Formula Ic: . [0030] In some

from:

AttyDktNo. R1119431100WO .

. [0032] In some

to Formula Id: , wherein R¹ is hydrogen or

[0033] In some embodiments, the compound of Formula Id has a structure selected from: AttyDktNo. R1119431100WO . [0034] In some

OH OH . [0035] In another aspect

a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient. For avoidance of doubt, reference herein to compounds of Formula I (e.g., with respect to compositions and methods of treatment) is intended to include compounds of each of Formulae Ia, Ib, Ic, and Id. [0036] In a further aspect 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 Formula I, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising said compound (or pharmaceutically acceptable salt of solvate thereof). [0037] In some embodiments, the bacterial infection is with Gram-positive, Gram-negative, aerobic, or facultative anaerobic bacteria. In some embodiments, the bacterial infection is a Gram-negative bacterial infection. In some embodiments, the bacterial infection is a Gram-positive bacterial infection. [0038] In some embodiments, the bacterial infection is with a member of the order of Enterobacterales. [0039] In some embodiments, the bacterial infection is infection with a Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, AttyDktNo. R1119431100WO Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Burkholderia or Enterococcus species. [0040] In some embodiments, the bacterial infection is infection with Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, Proteus mirabilis, Bacillus anthracis, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei or Yersinia pestis. [0041] In some embodiments, the bacterial infection is infection with an Escherichia spp., a Klebsiella spp., a Proteus spp., a Citrobacter spp., a Morganella spp., a Providencia spp., a Yersinia spp., an Enterobacter spp., a Salmonella spp., or a Serratia spp. [0042] In some embodiments, the bacterial infection is infection with a Moraxella spp., a Pseudomonas spp., an Acinetobacter spp., a Mycobacterium spp., a Staphylococcus spp., a Bacillus spp., a Francisella spp., or a Burkholderia spp. [0043] In some embodiments, the bacterial infection is a Gram-negative bacterial infection. In some embodiments, the Gram-negative bacterial infection is an infection with Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxidans. [0044] In some embodiments, the bacterial infection is caused by multidrug resistant (MDR) bacteria. [0045] In some embodiments, the bacterial infection is infection with an extended spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae. [0046] In some embodiments, the bacterial infection is caused by Staphylococcus aureus. In some embodiments, the Staphylococcus aureus is Methicillin-resistant Staphylococcus aureus (MRSA). [0047] The disclosure includes, without limitations, the following embodiments. [0048] Embodiment 1: A compound having a structure according to Formula I: , or a pharmaceutically

R¹ and R² are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, - S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R³ and R⁴ are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently AttyDktNo. R1119431100WO selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, - S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R⁵ is hydrogen or is C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, - OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and –CN; R⁶ is independently selected for each occurrence from the group consisting of hydrogen, C_{1- 10} alkyl, C_2-10 alkenyl, and C_2-10 alkynyl, wherein each of said C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl is optionally substituted with one or more substituents independently selected for each occurrence from halogen, -OH, -CN, -NO₂, and -NH₂; R⁷ is selected from the group consisting of hydrogen, -OR⁶, and -N(R⁶)2; R⁸ is selected from the group consisting of hydrogen and -OR⁶; R^a is a tetrahydrofuranyl ring or a hexahydropyranyl ring, connected via a glycosidic bond; R^b is a second tetrahydrofuranyl or hexahydropyranyl ring connected to R^a through a glycosidic bond; wherein each of R^a and R^b is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OR⁶, -N(R⁶)_2, and C_1-6 alkyl, wherein said C_1-6 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, C1-6 alkyl, -OR⁶, -N(R⁶)2, -NO2, and –CN; and R^c is hydrogen, C_1-6 alkyl, a protecting group, or has a structure according to Formula II: , wherein:

Q is NH, O, or S; n is an integer from 0 to 4; R⁹ is hydrogen or C_1-3 alkyl; R¹⁰ and R¹¹ are each selected independently for each occurrence from the group consisting of hydrogen, alkyl, halogen, and –OH; and R¹² is H, C_1-3 alkyl, or –C(=NH)NR^12aR^12b, wherein R^12a and R^12b are each independently selected from the group consisting of hydrogen and C_1-3 alkyl; or R⁹ and R¹², together with the atoms to which they are attached, form a heterocycloalkyl ring system comprising at least one N. [0049] Embodiment 2: The compound of embodiment 1, wherein R¹ and R² are each hydrogen. [0050] Embodiment 3: The compound of embodiment 1, wherein R¹ and R² are each methyl. [0051] Embodiment 4: The compound of embodiment 1, wherein one of R¹ and R² is hydrogen, and the other of R¹ and R² is methyl. AttyDktNo. R1119431100WO [0052] Embodiment 5: The compound of any one of embodiments 1 to 4, wherein R³ is hydrogen and R⁴ is methyl. [0053] Embodiment 6: The compound of any one of embodiments 1 to 4, wherein R¹, R² and R³ are hydrogen, and R⁴ is methyl. [0054] Embodiment 7: The compound of any one of embodiments 1 to 4, wherein R¹ and R³ are both hydrogen and R² and R⁴ are both methyl. [0055] Embodiment 8: The compound of any one of embodiments 1 to 7, wherein R⁵ is hydrogen. [0056] Embodiment 9: The compound of any one of embodiments 1 to 8, wherein R^c is selected from the group consisting of: . [0057] Embodiment 10: The wherein R^c is:

. [0058] Embodiment 11:

a structure according to Formula Ia: , wherein:

R¹³ and R¹⁴ are each independently selected from –OR⁶ and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of –OR⁶, N(R⁶)₂, and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, -OR⁶, -N(R⁶)₂, -NO₂, and –CN. [0059] Embodiment 12: The compound of embodiment 11, wherein: R¹³ is H or OH; and AttyDktNo. R1119431100WO R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl. [0060] Embodiment 13: The compound of embodiment 12, wherein R' and R'' are each hydrogen. [0061] Embodiment 14: The compound of embodiment 12, wherein R' is hydrogen and R'' is methyl. [0062] Embodiment 15: The compound of embodiment 12, wherein R' and R'' are each methyl. [0063] Embodiment 16: The compound of embodiment 12, wherein R¹³ is -OH and R¹⁴ is -CH₂OH. [0064] Embodiment 17: The compound of embodiment 11, having a structure according to Formula Ib: , wherein: R¹ is hydrogen or

R⁷ is NH₂ or OH; R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of -OH, -NH₂, and C_1-3 alkyl, said C_1-3 alkyl optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OH, -NH₂, -NO₂, and -CN. [0065] Embodiment 18: The compound of embodiment 17, wherein R⁷ is NH_2. [0066] Embodiment 19: The compound of embodiment 17, having a structure according to Formula Ic: . [0067] Embodiment 20:

selected from: AttyDktNo. R1119431100WO .

. [0069] Embodiment 22:

according to Formula Id: , wherein R¹ is hydrogen or

[0070] Embodiment 23: The compound of embodiment 22, having a structure selected from: AttyDktNo. R1119431100WO . [0071] Embodiment

OH OH . [0072] Embodiment 25:

of any one of embodiments 1-24, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, and at least one pharmaceutically acceptable excipient. [0073] Embodiment 26: A method for treating a bacterial infection in a subject in need thereof, the method comprising administering to the subject a compound of any one of embodiments 1-24, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or the pharmaceutical composition of embodiment 25. [0074] Embodiment 27: The method of embodiment 26, wherein the bacterial infection is with Gram- positive, Gram-negative, an aerobic, or facultative anaerobic bacteria. [0075] Embodiment 28: The method of embodiment 26, wherein the bacterial infection is with a member of the order of Enterobacterales. [0076] Embodiment 29: The method of embodiment 26, wherein the bacterial infection is with an Escherichia spp., a Klebsiella spp., a Proteus spp., a Citrobacter spp., a Morganella spp., a Providencia spp., a Yersinia spp., an Enterobacter spp., a Salmonella spp., or a Serratia spp. AttyDktNo. R1119431100WO [0077] Embodiment 30: The method of embodiment 26, wherein the bacterial infection is with a Moraxella spp., a Pseudomonas spp., an Acinetobacter spp., a Mycobacterium spp., a Staphylococcus spp., a Bacillus spp., a Francisella spp., or a Burkholderia spp. [0078] Embodiment 31: The method of embodiment 26, wherein the bacterial infection is a Gram-negative bacterial infection. [0079] Embodiment 32: The method of embodiment 26, wherein the Gram-negative bacterial infection is an infection with Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxidans, [0080] Embodiment 33: The method of embodiment 26, wherein the bacterial infection is caused by multidrug resistant (MDR) bacteria. [0081] Embodiment 34: The method of embodiment 26, wherein the bacterial infection is infection with an extended spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae. [0082] Embodiment 35: The method of embodiment 26, wherein the bacterial infection is caused by Staphylococcus aureus. [0083] Embodiment 36: The method of embodiment 35, wherein the Staphylococcus aureus is Methicillin- resistant Staphylococcus aureus (MRSA). [0084] These and other features, aspects, and advantages of the present disclosure will be apparent from a reading of the following detailed description. The present disclosure includes any combination of two, three, four or more features or elements set forth in this disclosure, regardless of whether such features or elements are expressly combined or otherwise recited in a specific example implementation described herein. This disclosure is intended to be read holistically such that any separable features or elements of the disclosure, in any of its aspects and example implementations, should be viewed as combinable, unless the context of the disclosure clearly dictates otherwise. [0085] It will therefore be appreciated that this Summary is provided merely for purposes of summarizing some example implementations so as to provide a basic understanding of some aspects of the disclosure. Accordingly, it will be appreciated that the above-described example implementations are merely examples and should not be construed to narrow the scope or spirit of the disclosure in any way. Other example implementations, aspects, and advantages will become apparent from the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS [0086] Having thus described aspects of the disclosure in the foregoing general terms, reference will now be made to the accompanying drawings, which are not necessarily drawn to scale. The drawings are exemplary only and should not be construed as limiting the disclosure. [0087] FIG.1 illustrates a non-limiting scheme for preparation of a compound of the disclosure having the structure of Ring A1 according to one embodiment. [0088] FIG.2 illustrates a non-limiting scheme for preparation of a compound of the disclosure having the structure of Ring A2 according to one embodiment. AttyDktNo. R1119431100WO DETAILED DESCRIPTION [0089] The present disclosure will now be described more fully hereinafter with reference to example embodiments thereof. Before describing several example embodiments of the technology, it is to be understood that the technology is not limited to the details of construction or process steps set forth in the following description. The technology is capable of other embodiments and of being practiced or being carried out in various ways. [0090] The following description sets forth numerous exemplary configurations, methods, parameters, and the like in order to provide a thorough understanding of various embodiments of the disclosure. It should be recognized, however, that such description is not intended as a limitation on the scope of the present disclosure but is instead provided as a description of exemplary embodiments. [0091] The present disclosure is generally directed to aminoglycoside compounds, including stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising said compounds, or stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof. The disclosure particularly relates to such compounds, stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof with antibacterial activity. Accordingly, compounds and pharmaceutical compositions of the disclosure may be useful for treating bacterial infections as well as medical conditions associated therewith. Compounds and pharmaceutical compositions of the disclosure may also be useful for treating difficult-to-treat resistance mechanisms as well as multidrug resistant (MDR) bacterial infections. [0092] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this disclosure belongs. All patents and publications referred to herein are incorporated by reference. Definitions [0093] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs. With respect to the terms used in this disclosure, the following definitions are provided. This application will use the following terms as defined below unless the context of the text in which the term appears requires a different meaning. [0094] The articles "a" and "an" as used in this disclosure may refer to one or 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. [0095] The term "and/or" as used in this disclosure may mean either "and" or "or" unless indicated otherwise. [0096] The term "about" used throughout this specification is used to describe and account for small fluctuations. For example, the term "about" can refer to less than or equal to ±10%, less than or equal to ±5%, less than or equal to ±2%, less than or equal to ±1%, less than or equal to ±0.5%, less than or equal to ±0.2%, less than or equal to ±0.1% or less than or equal to ±0.05%. All numeric values herein are modified by the AttyDktNo. R1119431100WO term "about," whether or not explicitly indicated. A value modified by the term "about" of course includes the specific value. For instance, "about 5.0" must include 5.0. [0097] Unless the context requires otherwise, throughout the present specification and claims, as used herein, the terms "including," "containing," and "comprising" are used in their open, non-limiting sense. [0098] "Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, and preferably having from one to fifteen carbon atoms (i.e., C₁- C₁₅ alkyl). In certain embodiments, an alkyl comprises one to thirteen carbon atoms (i.e., C₁-C₁₃ alkyl). In certain embodiments, an alkyl comprises one to eight carbon atoms (i.e., C₁-C₈ alkyl). In other embodiments, an alkyl comprises one to five carbon atoms (i.e., C₁-C₅ alkyl). In other embodiments, an alkyl comprises one to four carbon atoms (i.e., C₁-C₄ alkyl). In other embodiments, an alkyl comprises one to three carbon atoms (i.e., C1-C3 alkyl). In other embodiments, an alkyl comprises one to two carbon atoms (i.e., C1-C2 alkyl). In other embodiments, an alkyl comprises one carbon atom (i.e., C₁ alkyl). In other embodiments, an alkyl comprises five to fifteen carbon atoms (i.e., C₅-C₁₅ alkyl). In other embodiments, an alkyl comprises five to eight carbon atoms (i.e., C₅-C₈ alkyl). In other embodiments, an alkyl comprises two to five carbon atoms (i.e., C₂-C₅ alkyl). In other embodiments, an alkyl comprises three to five carbon atoms (i.e., C₃-C₅ alkyl). In certain embodiments, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl is attached to the rest of the molecule by a single bond. [0099] The term "C_x-y" when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain. For example, the term "C_1-6alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from 1 to 6 carbons. The term –C_x-yalkylene- refers to a substituted or unsubstituted alkylene chain with from x to y carbons in the alkylene chain. For example –C_1-6alkylene- may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any one of which is optionally substituted. [00100] "Alkoxy" refers to a radical bonded through an oxygen atom of the formula –o-alkyl, where alkyl is an alkyl chain as defined above. [00101] "Alkenyl" refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and preferably having from two to twelve carbon atoms (i.e., C₂-C₁₂ alkenyl). In certain embodiments, an alkenyl comprises two to eight carbon atoms (i.e., C₂-C₈ alkenyl). In certain embodiments, an alkenyl comprises two to six carbon atoms (i.e., C₂-C₆ alkenyl). In other embodiments, an alkenyl comprises two to four carbon atoms (i.e., C₂-C₄ alkenyl). The alkenyl is attached to the rest of the molecule by a single bond, 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. [00102] "Alkynyl" refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, and preferably having from two to twelve carbon atoms (i.e., C₂-C₁₂ alkynyl). In certain embodiments, an alkynyl comprises two to eight carbon AttyDktNo. R1119431100WO atoms (i.e., C₂-C₈ alkynyl). In other embodiments, an alkynyl comprises two to six carbon atoms (i.e., C₂-C₆ alkynyl). In other embodiments, an alkynyl comprises two to four carbon atoms (i.e., C₂-C₄ alkynyl). The alkynyl is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. [00103] The terms "C_x-yalkenyl" and "C_x-yalkynyl" refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively. The term –C_x-yalkenylene- refers to a substituted or unsubstituted alkenylene chain with from x to y carbons in the alkenylene chain. For example, –C_2-6alkenylene- may be selected from ethenylene, propenylene, butenylene, pentenylene, and hexenylene, any one of which is optionally substituted. An alkenylene chain may have one double bond or more than one double bond in the alkenylene chain. The term –Cx-yalkynylene- refers to a substituted or unsubstituted alkynylene chain with from x to y carbons in the alkenylene chain. For example, –C_2-6alkenylene- may be selected from ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any one of which is optionally substituted. An alkynylene chain may have one triple bond or more than one triple bond in the alkynylene chain. [00104] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation, and preferably having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, n-butylene, and the like. 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 may be through any two carbons within the chain. In certain embodiments, an alkylene comprises one to ten carbon atoms (i.e., C₁-C₈ alkylene). In certain embodiments, an alkylene comprises one to eight carbon atoms (i.e., C₁-C₈ alkylene). In other embodiments, an alkylene comprises one to five carbon atoms (i.e., C₁-C₅ alkylene). In other embodiments, an alkylene comprises one to four carbon atoms (i.e., C₁-C₄ alkylene). In other embodiments, an alkylene comprises one to three carbon atoms (i.e., C₁- C₃ alkylene). In other embodiments, an alkylene comprises one to two carbon atoms (i.e., C₁-C₂ alkylene). In other embodiments, an alkylene comprises one carbon atom (i.e., C₁ alkylene). In other embodiments, an alkylene comprises five to eight carbon atoms (i.e., C₅-C₈ alkylene). In other embodiments, an alkylene comprises two to five carbon atoms (i.e., C₂-C₅ alkylene). In other embodiments, an alkylene comprises three to five carbon atoms (i.e., C₃-C₅ alkylene). [00105] "Alkenylene" or "alkenylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond, and preferably having from two to twelve carbon atoms. 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 may be through any two carbons within the chain. In certain embodiments, an alkenylene comprises two to ten carbon atoms (i.e., C₂-C₁₀ alkenylene). In certain embodiments, an alkenylene comprises two to eight carbon atoms (i.e., C₂- C₈ alkenylene). In other embodiments, an alkenylene comprises two to five carbon atoms (i.e., C₂-C₅ AttyDktNo. R1119431100WO alkenylene). In other embodiments, an alkenylene comprises two to four carbon atoms (i.e., C₂-C₄ alkenylene). In other embodiments, an alkenylene comprises two to three carbon atoms (i.e., C₂-C₃ alkenylene). In other embodiments, an alkenylene comprises two carbon atom (i.e., C₂ alkenylene). In other embodiments, an alkenylene comprises five to eight carbon atoms (i.e., C₅-C₈ alkenylene). In other embodiments, an alkenylene comprises three to five carbon atoms (i.e., C₃-C₅ alkenylene). [00106] "Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, and preferably having from two to twelve carbon atoms. 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 may be through any two carbons within the chain. In certain embodiments, an alkynylene comprises two to ten carbon atoms (i.e., C₂-C₁₀ alkynylene). In certain embodiments, an alkynylene comprises two to eight carbon atoms (i.e., C₂-C₈ alkynylene). In other embodiments, an alkynylene comprises two to five carbon atoms (i.e., C₂-C₅ alkynylene). In other embodiments, an alkynylene comprises two to four carbon atoms (i.e., C₂-C₄ alkynylene). In other embodiments, an alkynylene comprises two to three carbon atoms (i.e., C₂-C₃ alkynylene). In other embodiments, an alkynylene comprises two carbon atom (i.e., C₂ alkynylene). In other embodiments, an alkynylene comprises five to eight carbon atoms (i.e., C₅-C₈ alkynylene). In other embodiments, an alkynylene comprises three to five carbon atoms (i.e., C3-C5 alkynylene). [00107] "Aryl" refers to a radical derived from an aromatic monocyclic or aromatic multicyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. The aromatic monocyclic or aromatic multicyclic hydrocarbon ring system contains only hydrogen and carbon and from five to eighteen carbon atoms, where at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) ^–electron system in accordance with the Hückel theory. The ring system from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin and naphthalene. [00108] "Aralkyl" refers to a radical of the formula -R^c-aryl where R^c is an alkylene chain as defined above, for example, methylene, ethylene, and the like. [00109] "Aralkenyl" refers to a radical of the formula –R^d-aryl where R^d is an alkenylene chain as defined above. "Aralkynyl" refers to a radical of the formula -R^e-aryl, where R^e is an alkynylene chain as defined above. [00110] "Carbocycle" refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon. Carbocycle may 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 may be selected from saturated, unsaturated, and aromatic rings. In some embodiments, the carbocycle is an aryl. In some embodiments, the carbocycle is a cycloalkyl. In some embodiments, the carbocycle is a cycloalkenyl. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as AttyDktNo. R1119431100WO valence permits, are included in the definition of carbocyclic. Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. [00111] "Cycloalkyl" refers to a stable fully saturated monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused or bridged ring systems, and preferably having from three to twelve carbon atoms. In certain embodiments, a cycloalkyl comprises three to ten carbon atoms. In other embodiments, a cycloalkyl comprises five to seven carbon atoms. The cycloalkyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. [00112] "Cycloalkenyl" refers to a stable unsaturated non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused or bridged ring systems, preferably having from three to twelve carbon atoms and comprising at least one double bond. In certain embodiments, a cycloalkenyl comprises three to ten carbon atoms. In other embodiments, a cycloalkenyl comprises five to seven carbon atoms. The cycloalkenyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. [00113] "Cycloalkylalkyl" refers to a radical of the formula –R^c-cycloalkyl where R^c is an alkylene chain as described above. [00114] "Cycloalkylalkoxy" refers to a radical bonded through an oxygen atom of the formula –O- R^c-cycloalkyl where R^c is an alkylene chain as described above. [00115] "Halo" or "halogen" refers to halogen substituents such as bromo, chloro, fluoro and iodo substituents. [00116] As used herein, the term "haloalkyl" or "haloalkane" refers to an alkyl radical, as defined above, that is substituted by one or more halogen radicals, for example, trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. In some embodiments, the alkyl part of the fluoroalkyl radical is optionally further substituted. Examples of halogen substituted alkanes ("haloalkanes") include halomethane (e.g., chloromethane, bromomethane, fluoromethane, iodomethane), di-and trihalomethane (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 combinations of alkanes (or substituted alkanes) and halogens (e.g., Cl, Br, F, I, etc.). When an alkyl group is substituted with more than one halogen radicals, each halogen may be independently selected e.g., 1-chloro,2-fluoroethane. [00117] "Fluoroalkyl" refers to an alkyl radical, as defined above, that is substituted by one or more fluoro radicals, for example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. [00118] "Heterocycle" refers to a saturated, unsaturated or aromatic ring comprising one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered AttyDktNo. R1119431100WO monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic heterocycle may be selected from saturated, unsaturated, and aromatic rings. In some embodiments, the heterocycle is a heteroaryl. In some embodiments, the heterocycle is a heterocycloalkyl. "Heterocyclene" refers to a divalent heterocycle linking the rest of the molecule to a radical group [00119] "Heterocycloalkyl" refers to a stable 3- to 12-membered non-aromatic ring radical that comprises two to twelve carbon atoms and at least one heteroatom wherein each heteroatom may be selected from N, O, Si, P, B, and S atoms. The heterocycloalkyl 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 rest of the molecule through any atom of the heterocycloalkyl, valence permitting, such as any carbon or nitrogen atoms of the heterocycloalkyl. 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. [00120] "Heterocycloalkylalkyl" refers to a radical of the formula –R^c-heterocycloalkyl where R^c 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. [00121] "Heteroaryl" or "aromatic heterocycle" refers to a radical derived from a 3- to 12-membered aromatic ring radical that comprises one to eleven carbon atoms and at least one heteroatom wherein each heteroatom may be selected from N, O, and S. As used herein, the heteroaryl ring may be selected from monocyclic or bicyclic and fused or bridged ring systems rings wherein at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) ^–electron system in accordance with the Hückel theory. The heteroatom(s) 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 rest of the molecule through any atom of the heteroaryl, valence permitting, such as a carbon or nitrogen atom of the heteroaryl. Examples of heteroaryls include, but are not limited to, 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, 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, AttyDktNo. R1119431100WO isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, 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[2,3-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). An "X-membered heteroaryl" refers to the number of endocylic atoms, i.e., X, in the ring. For example, a 5-membered heteroaryl ring or 5-membered aromatic heterocycle has 5 endocyclic atoms, e.g., triazole, oxazole, thiophene, etc. [00122] "Heteroarylalkyl" refers to a radical of the formula –R^c-heteroaryl, where R^c 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. [00123] "Isomers" are different compounds that have 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 where appropriate. "Diastereoisomers" or "diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. The absolute stereochemistry is specified according to the Cahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified by either R or S. Resolved compounds whose absolute configuration is unknown can be designated (+) or (-) depending on the direction (dextro- or levorotatory) in which they rotate plane polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms, the asymmetric centers of which can be defined, in terms of absolute stereochemistry, as (R) or (S). The present chemical entities, pharmaceutical compositions and methods are meant to include all such possible stereoisomers, including racemic mixtures, optically pure forms, mixtures of diastereomers and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents or resolved using conventional techniques. The optical activity of a compound can be analyzed via any suitable method, including but not limited to chiral chromatography and polarimetry, and the degree of predominance of one stereoisomer over the other isomer can be determined. [00124] In certain embodiments, the compounds of the disclosure may contain asymmetric or chiral centers, and, therefore, exist in different stereoisomeric forms. The term "stereoisomers" may refer to the set of compounds which have the same number and type of atoms and share the same bond connectivity between those atoms but differ in three-dimensional structure. The term "stereoisomer" may refer to any member of AttyDktNo. R1119431100WO this set of compounds. For instance, a stereoisomer may be an enantiomer or a diastereomer. It is intended that all stereoisomeric forms of the compounds of the disclosure as well as mixtures thereof, including racemic mixtures, form part of the present disclosure. [00125] When stereochemistry is not specified, certain molecules described herein include isomers, such as enantiomers and diastereomers, mixtures of enantiomers, including racemates, mixtures of diastereomers, and other mixtures thereof, to the extent they can be made by one of ordinary skill in the art by routine experimentation. In certain embodiments, the single enantiomers or diastereomers, i.e., optically active forms, can be obtained by asymmetric synthesis or by resolution of the racemates or mixtures of diastereomers. Resolution of the racemates or mixtures of diastereomers, if possible, can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example, a chiral high-pressure liquid chromatography (HPLC) column. Furthermore, a mixture of two enantiomers enriched in one of the two can be purified to provide further optically enriched form of the major enantiomer by recrystallization and/or trituration. [00126] In certain embodiments, the chiral centers of the present disclosure may have the R- or S- configuration as defined by the IUPAC 1974 Recommendations. [00127] The term "amino" as used herein refers to –NH₂. [00128] The term "hydroxy" or "hydroxyl" as used herein refers to –OH. [00129] The term "oxo" as used herein refers to an "=O" group. It can also be abbreviated herein as C(O) or as C=O. [00130] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., NH, of the structure. It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. In certain embodiments, substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group. As used herein, the term "substituted" is contemplated 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 non-aromatic 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. [00131] In some embodiments, substituents may include any substituents described herein, for example: halogen, hydroxy, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO₂), imino (=N-H), oximo (=N-OH), hydrazino(=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)R^a, -R^b-C(O)OR^a, AttyDktNo. R1119431100WO -R^b-C(O)N(R^a)₂, -R^b-O-R^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)_tR^a (where t is 1 or 2), -R^b-S(O)_tR^a (where t is 1 or 2), -R^b-S(O)_tOR^a (where t is 1 or 2), and -R^b-S(O)_tN(R^a)₂ (where t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl any of which may be optionally substituted by alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO₂), imino (=N-H), 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)R^a, -R^b-C(O)OR^a, -R^b-C(O)N(R^a)₂, -R^b-O-R^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)_tR^a (where t is 1 or 2), -R^b-S(O)_tR^a (where t is 1 or 2), -R^b-S(O)_tOR^a (where t is 1 or 2) and -R^b-S(O)_tN(R^a)₂ (where t is 1 or 2); wherein each R^a is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, wherein each R^a, valence permitting, may be optionally substituted with alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO₂), imino (=N-H), 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)R^a, -R^b-C(O)OR^a, -R^b-C(O)N(R^a)₂, -R^b-O-R^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)_tR^a (where t is 1 or 2), -R^b-S(O)_tR^a (where t is 1 or 2), -R^b-S(O)_tOR^a (where t is 1 or 2) and -R^b-S(O)_tN(R^a)₂ (where t is 1 or 2); and wherein each R^b is 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. [00132] The term "radical of a compound" as used herein refers to a structure derived from a parent compound by removal of one or more atoms, e.g., hydrogen atoms. In one embodiment, a "radical of a compound" is a monovalent radical derived from the removal of one hydrogen atom from the parent compound. [00133] It is to be understood that certain radical naming conventions can include either a mono-radical or a di-radical, depending on the context. For example, where a substituent requires two points of attachment to the rest of the molecule, it is understood that the substituent is a di-radical. For example, a substituent identified as alkyl that requires two points of attachment includes di-radicals such as -CH₂-, -CH₂CH₂-, - CH₂CH(CH₃)CH₂-, and the like. Other radical naming conventions clearly indicate that the radical is a di- radical such as "alkylene," "alkenylene," "arylene," and the like. [00134] Wherever a substituent is depicted as a di-radical (i.e., has two points of attachment to the rest of the molecule), it is to be understood that the substituent can be attached in any directional configuration unless otherwise indicated. [00135] A "tautomer" refers to a molecule wherein a proton shift from one atom of a molecule to another atom of the same molecule is possible. The compounds presented herein, in certain embodiments, exist as tautomers. In circumstances where tautomerization is possible, a chemical equilibrium of the tautomers will exist. The exact ratio of the tautomers depends on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibrium include: AttyDktNo. R1119431100WO OH O O OH N N NH [00136] As

no substituents beyond the moiety recited (e.g., where valency is satisfied by hydrogen). [00137] "Stable compound" and "stable structure" may indicate 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. [00138] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. [00139] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, 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. [00140] The phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" as used herein means 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 injurious to the patient. Some examples of materials which 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 carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene AttyDktNo. R1119431100WO glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, 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 solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations. [00141] The term "carrier," as used in this disclosure, may encompass carriers, excipients, and diluents and may mean a material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting a pharmaceutical agent, such as one or more compounds, or pharmaceutically acceptable salts, solvates (e.g., hydrates), isomers (e.g., stereoisomers), and tautomers thereof, of the disclosure, from one organ, or portion of the body, to another organ, or portion of the body of a subject. Carriers should be selected on the basis of compatibility and the release profile properties of the desired dosage form. Exemplary carrier materials may include, e.g., adjuvants, binders, suspending agents, disintegration agents, filling 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 without limitation any adjuvant, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals. [00142] The terms "pharmaceutically acceptable" or "pharmacologically acceptable" may refer to a material which is not biologically, or otherwise, undesirable—the material may be administered to an individual without causing any substantially undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained. [00143] A "pharmaceutical composition" may refer to a formulation of a compound of the disclosure and a medium generally accepted in the art for the delivery of the biologically active compound to a subject, e.g., mammals or humans. Such a medium may include all pharmaceutically acceptable carriers therefor. [00144] The terms "subject," "individual," and "patient" may be used interchangeably and refer to humans, as well as non-human mammals (e.g., non-human primates, canines, equines, felines, porcines, bovines, ungulates, lagomorphs, and the like). In various embodiments, the subject can be a human (e.g., adult male, adult female, adolescent male, adolescent female, male child, female child) under the care of a physician or other health worker in a hospital, as an outpatient, or other clinical context. In certain embodiments, the subject may not be under the care or prescription of a physician or other health worker. [00145] As used herein, the phrase "a subject in need thereof" refers to a subject, as described infra, that suffers from, or is at risk for, a pathology to be prophylactically or therapeutically treated with a compound or salt described herein. [00146] As used herein, "agent" or "biologically active agent" refers to a biological, pharmaceutical, or chemical compound or other moiety. Non-limiting examples include an organic or inorganic molecule, a AttyDktNo. R1119431100WO peptide, a protein, a peptide nucleic acid (PNA), an oligonucleotide (including e.g., aptamer and polynucleotides), an antibody, an antibody derivative, antibody fragment, a vitamin derivative, a carbohydrate, a toxin, a branched chain amino acid in free amino acid form or metabolite thereof, or a chemotherapeutic compound. Various compounds can be synthesized, for example, small molecules and oligomers (e.g., oligopeptides and oligonucleotides), and synthetic organic compounds based on various core structures. In addition, various natural sources can provide compounds for screening, such as plant or animal extracts, and the like. A skilled artisan can readily recognize that there is no limit as to the structural nature of the agents. [00147] 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 certain embodiments, oral routes of administering a composition can be used. The term "effective amount" or "therapeutically effective amount" refers to that amount of a compound or salt described herein that is sufficient to effect the intended application including but not limited to disease treatment, as defined below. The therapeutically effective amount may vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. The term can also apply to a dose that can induce a particular response in target cells, e.g., reduction of proliferation or down regulation of activity of a target protein. The specific dose can vary depending on the particular compounds chosen, the dosing regimen to be followed, whether it is administered in combination with other compounds, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried. [00148] As used herein, "treatment" or "treating" refers to an approach for obtaining beneficial or desired results with respect to a disease, disorder, or medical condition including, but not limited to, a therapeutic benefit and/or a prophylactic benefit. In certain embodiments, treatment or treating involves administering a compound or composition disclosed herein to a subject. A therapeutic benefit may include the eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit may be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder, such as observing an improvement in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. In certain embodiments, for prophylactic benefit, the compositions are administered to a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made. Treating can include, for example, reducing, delaying or alleviating the severity of one or more symptoms of the disease or condition, or it can include reducing the frequency with which symptoms of a disease, defect, disorder, or adverse condition, and the like, are experienced by a patient. Treating can be used herein to refer to a method that results in some level of treatment or amelioration of the disease or condition and can contemplate a range of results directed to that end, including but not restricted to prevention of the condition entirely. AttyDktNo. R1119431100WO [00149] In certain embodiments, the term "prevent" or "preventing" as related to a disease or disorder may refer to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in the treated sample relative to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition relative to the untreated control sample. [00150] The term "prodrug" is intended to encompass compounds which, under physiologic conditions, are converted into the therapeutically pharmaceutical agents of the present disclosure. A common method for making a prodrug is to include one or more selected moieties which are hydrolyzed under physiologic conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids and esters of phosphonates) are preferred prodrugs of the present disclosure. [00151] The terms "treat," "treating" or "treatment," as used herein, may include alleviating, abating or ameliorating a disease or condition symptoms, preventing additional symptoms, ameliorating or preventing the underlying causes of symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition either prophylactically and/or therapeutically. [00152] The terms "effective amount" or "therapeutically effective amount" when used in connection with one or more compounds or pharmaceutical compositions may refer to a sufficient amount of the one or more compounds or pharmaceutical compositions to provide the desired biological result. That result can be 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 may be the amount of the pharmaceutical composition comprising one or more compounds, or pharmaceutically acceptable salts, solvates (e.g., hydrates), isomers (e.g., stereoisomers), and tautomers thereof, as disclosed herein required to provide a clinically significant decrease in a disorder. An appropriate "effective amount" in any individual case may be determined by one of ordinary skill in the art using routine experimentation. [00153] In certain embodiments, the terms "disease" and "condition" may be used interchangeably or may be different in that the particular malady or condition may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognized as a disease but only as an undesirable condition or syndrome, wherein a more or less specific set of symptoms have been identified by clinicians. [00154] The terms "antibacterial agent," "antibiotic," or "antibacterial compound" are used interchangeably herein and may refer to agents or compounds that have either bactericidal or bacteriostatic activity. The term "inhibiting the growth" may indicate that the rate of increase in the numbers of a population of a particular bacterium is reduced. Thus, the term may include situations in which the bacterial population increases but at a reduced rate, as well as situations where the growth of the population is stopped, as well as situations where the numbers of the bacteria in the population are reduced or the population even eliminated. The activity of antibacterial agents or compounds is not necessarily limited to bacteria but may also encompass general antimicrobial activity against parasites or fungi or general antiviral activity against viruses. AttyDktNo. R1119431100WO Compounds of the Disclosure [00155] Generally, the present disclosure provides aminoglycoside compounds which may be characterized as having antibacterial activity. [00156] In one aspect is provided a compound having a structure according to Formula I: , or a pharmaceutically 1 ²

R and R are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)2, -N(R⁶)2, -S(O)R⁶, -S(O)2R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R³ and R⁴ are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R⁵ is hydrogen or is C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, - OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and –CN; R⁶ is independently selected for each occurrence from the group consisting of hydrogen, C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl, wherein each of said C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl is optionally substituted with one or more substituents independently selected for each occurrence from halogen, -OH, - CN, -NO2, and -NH2; R⁷ is selected from the group consisting of hydrogen, -OR⁶, and -N(R⁶)₂; R⁸ is selected from the group consisting of hydrogen and -OR⁶; R^a is a tetrahydrofuranyl ring or a hexahydropyranyl ring, connected via a glycosidic bond; R^b is a second tetrahydrofuranyl or hexahydropyranyl ring connected to R^a through a glycosidic bond; wherein each of R^a and R^b is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OR⁶, -N(R⁶)_2, and C_1-6 alkyl, wherein said C_1-6 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and R^c is hydrogen, C_1-6 alkyl, a protecting group, or has a structure according to Formula II: AttyDktNo. R1119431100WO , wherein: Q is NH, O, or S;

n is an integer from 0 to 4; R⁹ is hydrogen or C1-3 alkyl; R¹⁰ and R¹¹ are each selected independently for each occurrence from the group consisting of hydrogen, alkyl, halogen, and –OH; and R¹² is H, C_1-3 alkyl, or –C(=NH)NR^12aR^12b, wherein R^12a and R^12b are each independently selected from the group consisting of hydrogen and C_1-3 alkyl; or R⁹ and R¹², together with the atoms to which they are attached, form a heterocycloalkyl ring system comprising at least one N. [00157] In some embodiments, R^a is a tetrahydrofuranyl ring connected via a glycosidic bond; and R^b is a hexahydropyranyl ring connected to R^a through a glycosidic bond, wherein each of R^a and R^b is substituted with one or more substituents independently selected for each occurrence from the group consisting of -OR⁶, -N(R⁶)_2, and C_1-6 alkyl, wherein said C_1-6 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, - NO₂, and –CN. Such compounds possess the four-ring system of the aminoglycosides. Accordingly, in some embodiments, compounds of Formula I (including Formula Ia, Ib, Ic, and Id) may be referred to herein as "ABCD compounds" in reference to the four-ring (aminoglycoside) system comprising the compounds, where the A, B, C, and D rings are as identified below. Conventional numbering is applied herein to such ring systems. For simplicity, the conventional numbering is illustrated below in reference to a simplified, non- limiting ABCD ring scaffold ABCD' (i.e., having several substituents removed relative to Formula I for ease of visualization): . [00158] The structure of format as shown below:

AttyDktNo. R1119431100WO . [00159] In some R¹ and R² are each methyl. In some embodiments, ^{1 2}

R and R is methyl. In some embodiments, R³ is hydrogen and R⁴ is methyl. In some embodiments, R¹, R² and R³ are hydrogen, and R⁴ is methyl. In some embodiments, R¹ and R³ are both hydrogen and R² and R⁴ are both methyl. Accordingly, in some embodiments, the A ring has a structure according to one of: H₂N H NH N O ^{O H} O _O H O _O [00160] In

[00161] In specific embodiments, the A ring has a structure according to one of:

AttyDktNo. R1119431100WO H₂N H NH O ^O H O _O . [00162] In some

. [00163] In some

. [00164] In some

to Formula Ia: , wherein:

R¹ to R¹² and R^c are each as described above; R¹³ and R¹⁴ are each independently selected from –OR⁶ and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and AttyDktNo. R1119431100WO R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of –OR⁶, N(R⁶)₂, and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, -OR⁶, -N(R⁶)₂, -NO₂, and –CN. [00165] In some embodiments, R¹³ is H or OH. [00166] In some embodiments, R² and R³ are hydrogen; R⁴ is methyl; R⁸ is OH; and R¹³ is OH. Accordingly, in some embodiments, the compound of Formula I has a structure according to Formula Ib: , wherein:

R¹ is hydrogen or R⁵ is hydrogen or is C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, - OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and –CN; R⁶ is independently selected for each occurrence from the group consisting of hydrogen, C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl, wherein each of said C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl is optionally substituted with one or more substituents independently selected for each occurrence from halogen, -OH, -CN, -NO₂, and -NH₂; R⁷ is NH2 or OH; R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of -OH, -NH₂, and C_1-3 alkyl, said C_1-3 alkyl optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OH, -NH₂, -NO₂, and CN. [00167] In some embodiments, R' and R'' are each hydrogen. In some embodiments, R' is hydrogen and R'' is methyl. In some embodiments, R' and R'' are each methyl. In some embodiments, R¹⁴ is -CH₂OH. [00168] In some embodiments, R⁷ is NH₂. [00169] In some embodiments, R^c is selected from the group consisting of: .

AttyDktNo. R1119431100WO [00170] In some embodiments, R^c is: . [00171] In some

R¹⁶ is OH; R¹⁷ is OH; and R¹⁸ is CH₂NH₂. Accordingly, in some embodiments, the compound of Formula I has a structure according to Formula Ic: . [00172] In some from:

.

.

AttyDktNo. R1119431100WO [00174] In other embodiments, R² and R³ are hydrogen; R⁴ is methyl; R⁷ is NH2; R¹⁴ is -CH2OH; R¹⁵ is NH₂; R¹⁶ is OH; R¹⁷ is OH; and R¹⁸ is CH₂NH₂. Accordingly, in some embodiments, the compound of Formula I has a structure according to Formula Id: OH H N N ^c R¹ ₂ HR , wherein R¹ is [00175] In some

[00176] In some embodiments, R¹ is hydrogen. Accordingly, in some embodiments, the compound of Formula Id has a structure selected from: .

AttyDktNo. R1119431100WO Isotopes and Isotopically Labeled Compounds [00178] The compounds described herein may exhibit their 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 the atomic mass or mass number 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. Accordingly, reference to a certain element is meant to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, it also includes isotopes thereof. For example, hydrogen has three naturally occurring isotopes, denoted ¹H (protium), ²H (deuterium), and ³H (tritium). Protium is the most abundant isotope of hydrogen in nature. Enriching for deuterium may afford certain therapeutic advantages, such as increased in vivo half-life and/or exposure, or may provide a compound useful for investigating in vivo routes of drug elimination and metabolism. Isotopically enriched compounds may be prepared by conventional techniques well known to those skilled in the art. [00179] The compounds described herein further include all pharmaceutically acceptable isotopically labeled compounds. An "isotopically" or "radio-labeled" compound may be a compound where one or more atoms are replaced or substituted by an atom having an atomic mass or mass number 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 hydrogen atoms are replaced or substituted by one or more deuterium or tritium. [00180] Certain isotopically labeled compounds of this disclosure, for example, those incorporating a radioactive isotope, may be useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, i.e., ³H, and carbon 14, i.e., ¹⁴C, may be particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium, i.e., ²H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. In some embodiments, the compound comprises at least one deuterium atom. For example, one or more hydrogen atoms in a compound of the present disclosure can be replaced or substituted by 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 in compounds described herein include but are not limited to ²H (also written as D for deuterium), ³H (also written as T for tritium), ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ¹⁸F, ³⁵S, ³⁶Cl, ⁸²Br, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I, and ¹³¹I. Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O, and ¹³N, can be useful in Positron Emission Topography (PET) studies. [00181] Isotopically labelled versions of the compounds disclosed herein can generally be prepared by following procedures analogous to those disclosed in the Schemes and/or in the Examples herein, by substituting an appropriate isotopically labelled reagent for a non-isotopically labelled reagent. [00182] The disclosure herein is also meant to encompass the in vivo metabolic products of the disclosed compounds. Such products may result from, for example, the oxidation, reduction, hydrolysis, amidation, AttyDktNo. R1119431100WO esterification, and the like of the administered compound, primarily due to enzymatic processes. Accordingly, the disclosure may include compounds produced by a process comprising administering a compound of this 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 disclosure in a detectable dose to subject, such as rat, mouse, guinea pig, monkey, or to human, allowing sufficient time for metabolism to occur, and isolating its conversion products from the urine, blood or other biological samples. Salts and Solvates [00183] The present disclosure further provides pharmaceutically acceptable salts, solvates (e.g., hydrates), and combinations thereof of any of the compounds as disclosed herein. The use of the terms "salt," "hydrate," "solvate," and the like, is intended to equally apply to the salt, hydrate, or solvate of enantiomers, diastereomers, isomers, stereoisomers, rotamers, tautomers, positional isomers, or racemates of the disclosed compounds. [00184] The term "salt" or "pharmaceutically acceptable salt" refers to salts derived from a variety of organic and inorganic counter ions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids 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 can 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 such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is chosen from ammonium, potassium, sodium, calcium, and magnesium salts. In some embodiments, the "pharmaceutically acceptable salts" may include, e.g., water-soluble and water-insoluble salts, such as the acetate, amsonate (4,4- diaminostilbene-2,2-disulfonate), benzenesulfonate, benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fiunarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, sethionate, lactate, lactobionate, laurate, magnesium, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, 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- AttyDktNo. R1119431100WO toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate salts. [00185] Compounds of the present invention also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof. [00186] Compounds of the disclosure, including their stereoisomers and tautomers, as well as salts of any thereof, may exist as solvates. Often crystallizations produce a solvate of the compound of the disclosure. As used herein, the term "solvate" may refer to an aggregate that comprises one or more molecules of a compound of the disclosure with one or more molecules of solvent. The solvent may be water, in which case the solvate may be a hydrate. Alternatively, the solvent may be an organic solvent. Thus, the compounds and salts of the present disclosure may exist as a hydrate, including a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compound of the disclosure may be true solvates, while in other cases the compound of the disclosure may merely retain adventitious water or be a mixture of water plus some adventitious solvent. Preparation of Compounds of the Disclosure [00187] Compounds of the present disclosure having an ABCD ring system as described herein above (e.g., compounds of Formula I) may 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 in conjunction with the guidance provided herein. In the schemes described below and provided in the Figures, it is understood that protecting groups for sensitive or reactive groups may be employed where necessary in accordance with general principles or chemistry in accordance with the guidance 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,” Third edition, Wiley, New York 1999). These groups may be removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art based on the detailed teaching provided herein. The selection processes, as well as the reaction conditions and order of their execution, shall be consistent with the present disclosure. [00188] Generally, the methods of preparing compounds of the present disclosure having an ABCD ring system comprise combinations of reactions and conditions that can provide certain novel intermediate compounds. Scheme 1 illustrates a general, representative, non-limiting strategy for preparation of compounds having an ABCD ring system (e.g., compounds of Formula I). AttyDktNo. R1119431100WO Scheme 1

[00189]

to Scheme 1 is discussed further herein below. The transformations in Scheme 1 can be performed through chemical reactions using standard synthetic chemistry procedures and practices optionally using reagents such as catalysts including, but not limited to, Pd reagents, chiral ligands, and enzymes. Preparation of Ring A [00190] Preparation of compounds having the ABCD ring system (e.g., compounds of Formula I) according to Scheme 1 utilizes a Ring A compound. In some embodiments, the Ring A compound has the general structure (A): , wherein: X¹ is an amine, an amine

group; X² is an amine, an amine precursor, or a protected amino group; Pg is a protecting group; and R¹ and R² are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted. [00191] In some embodiments, R¹ and R² are each hydrogen. In some embodiments, one of R¹ and R² is hydrogen, and the other is methyl. [00192] In some embodiments, X¹ is azide (N₃). In some embodiments, X¹ is –N(Bn)(Cbz). In some embodiments, X¹ is –N(CH₃)(Cbz). [00193] In some embodiments, X² is azide (N₃). [00194] In some embodiments, Pg is benzyl. [00195] In some embodiments, the Ring A compound has the structure of Formula A1 (i.e., Ring A1): AttyDktNo. R1119431100WO (Ring A1). [00196] An exemplary, non-limiting

of Ring A1 is illustrated in FIG. 1 and is discussed in greater detail herein. Generally, Ring A1 may be prepared starting from (3R,4R,5S,6R)-3-amino- 6-(hydroxymethyl)tetrahydro-2H-pyran-2,4,5-triol (D-glucosamine; 1) using combinations of protection, deprotection, and substitution reactions in sequence. With reference to FIG. 1, in some embodiments, D- glucosamine 1 is N-protected with a suitable protecting group, for example, phthalimide, to form 2, followed by protection of the hydroxyl groups as esters, for example, acetates to form 3. Suitable reagents include, for example, phthalic anhydride under basic conditions, followed by treatment with an acylating reagent such as acetic anhydride and an amine base. The anomeric center may then be protected with, for example, a thiol group, such as with thiophenol, thiocresol, or the like to form 4. Suitable reagents include, for example, p- thiocresol in the presence of a Lewis acid such as boron trifluoride etherate. [00197] In some embodiments, the ester (e.g., acetate) protecting groups are removed with, for example, sodium methoxide in methanol to form 5. [00198] In some embodiments, the 5-hydroxy and 6-hydroxymethyl groups are protected as an acetal, for example, a benzaldehyde acetal to form 6. One suitable reagent for such a transformation is benzaldehyde dimethyl acetal in the presence of an acid catalyst. [00199] In some embodiments, the 4-hydroxy group is then removed. A particularly suitable means of removal is reductive elimination of a thiocarbonate under radical conditions. For example, in some embodiments, the 4-hydroxy group is converted to a phenythiocarbonate with e.g., phenyl chlorothionocarbonate to form 7, followed by radical reduction with, for example tributyltin hydride and an initiator such as azobisisobutyronitrile to form 8. [00200] In some embodiments, the nitrogen protecting group (e.g., phthalimide) is removed. Suitable phthalimide removal conditions include, but are not limited to, treatment with a nucleophilic diamine such as ethylenediamine to form 9. In some embodiments, the liberated amino group is then protected. One particularly suitable form of protection is as the azide. Formation of an azide from an amine may be performed, for example, by treatment with a reagent such as trifluoromethanesulfonyl azide in the presence of a copper catalyst to form 10. [00201] In some embodiments, the 5-hydroxy and 6-hydroxymethyl groups are deprotected to form 11. For example, in some embodiments, the protecting group is an acetal, and the acetal is cleaved under acidic conditions. [00202] In some embodiments, the 6-hydroxymethyl group is converted to a protected amine, such as an azide. In some embodiments, this conversion is performed by deprotecting the protected 6-hydroxymethyl group, converting the hydroxyl group to a leaving group, and displacing the leaving group with azide to form 12. In some embodiments, the hydroxymethyl group is converted to the azide under Mitsunobu conditions AttyDktNo. R1119431100WO (diphenylphosphoryl azide and diisopropyl azodicarboxylate in the presence of a phosphine, such as triphenylphosphine). [00203] The anomeric hydroxyl group may then be revealed. In embodiments where the anomeric position is protected as a thioether, the thioether (e.g., p-tolylthio) may be treated with a halogenating agent such as N- bromosuccinimide, to provide Ring A1. [00204] In some embodiments, the Ring A compound has the structure of Formula A2 (i.e., Ring A2): (Ring A2). [00205] An exemplary scheme for

A2 is illustrated in FIG.2. Referring to FIG.2, in some embodiments, intermediate 11 (from FIG. 1) is oxidized to the corresponding aldehyde (13) under suitable conditions. One particularly suitable oxidizing reagent is Dess-Martin periodinane. [00206] In some embodiments, the aldehyde of 13 is converted to a chiral sulfinylimine with, e.g., (R)-2- methylpropane-2-sulfinamide in the presence of a Lewis acid dehydrating agent, such as copper sulfate to form 14. The sulfinylimine 14 may then be treated with a suitable organometallic reagent, such as methyl magnesium bromide, to give a chiral sulfinamide (15). [00207] In some embodiments, the resulting amine nitrogen is then protected with a suitable protecting group, e.g., benzyl by alkylation to form 16. The sulfinamide chiral auxiliary may then be cleaved, for example, under acidic conditions, and the resulting amino group protected with a suitable protecting group, for example, as the benzyl carbamate (17). [00208] In some embodiments, the masked anomeric hydroxyl group of 17 is then revealed. In some embodiments, wherein the anomeric position is protected as a thioether, the thioether (e.g., p-tolylthio) may be treated with a halogenating agent such as N-bromosuccinimide, to provide Ring A2. [00209] In some embodiments, the Ring A compound has the structure of Formula A3 (i.e., Ring A3): (Ring A3).

Ring A3 may be prepared discussed with respect to FIG.2 but using the S- antipode of 2-methylpropane-2-sulfinamide. [00210] In some embodiments, the Ring A compound has the structure of Formula A4 or A5 (i.e., Ring A4 or Ring A5, respectively): AttyDktNo. R1119431100WO

. Ring A4 may be prepared according to the general scheme provided for preparation of Ring A2, discussed with respect to FIG.2, but replacing the benzyl N-protecting group with methyl. Ring A5 may be prepared similarly to Ring A4 but using the S-antipode of 2-methylpropane-2-sulfinamide. Preparation of Ring B [00211] Preparation of compounds having the ABCD ring system (i.e., compounds of Formula I) according to Scheme 1 utilizes a Ring B compound. In some embodiments, the Ring B compound has the general structure: OR X X ; wherein:

X is an amine, an precursor, or a amino group; and R is a protecting group. [00212] In some embodiments, X is NH(Cbz) and R is Ac. Accordingly, in some embodiments, the B-Ring compound has the structure of Formula B1 (i.e., Ring B1): OAc CbzHN NHCbz (Ring B1). Ring B1 may be prepared

by hydrolysis of the guanidine groups, N- protection, O-protection, and hydrolysis of the glycosyl linkage. Procedures for the preparation of such Ring B compounds are reported in, for example, International Patent Application Publication Nos. WO2019194858 and WO2019046126, each of which is incorporated by reference herein with respect to the teachings regarding synthesis of Ring B1. Preparation of Ring CD [00213] Preparation of compounds having the ABCD ring system (i.e., compounds of Formula I) according to Scheme 1 utilizes a Ring CD compound. In some embodiments, the Ring CD compound has the structure of Formula CD1 (i.e., Ring CD1): AttyDktNo. R1119431100WO HO OAc O 3

. Ring CD-1 may be prepared according to the procedure reported in, for example, International Patent Application Publication No. WO2019/194858 previously incorporated by reference. [00214] In some embodiments, the Ring CD compound has the structure of Formula CD2 (i.e., Ring CD2): . Ring CD-2 may be prepared

procedure reported in, for example, International Patent Application Publication No. WO2019/194858 previously incorporated by reference. Preparation of Ring ABCD (Compounds of Formula Ia, Ib, Ic, and Id) [00215] Provided herein are compounds of Formula I having an ABCD ring system. In some embodiments the compound of Formula I having an ABCD ring system has a structure according to Formula Ia: OH R⁷ NHR^c , wherein:

R¹ and R² are each group hydrogen and C_1-4 alkyl, wherein said C1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, - S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; AttyDktNo. R1119431100WO R³ and R⁴ are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, - S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R⁵ is hydrogen or is C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, - OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and –CN; R⁶ is independently selected for each occurrence from the group consisting of hydrogen, C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl, wherein each of said C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl is optionally substituted with one or more substituents independently selected for each occurrence from halogen, -OH, -CN, -NO₂, and -NH₂; R⁷ is selected from the group consisting of hydrogen, -OR⁶, and -N(R⁶)₂; R⁸ is selected from the group consisting of hydrogen and -OR⁶; R^c is hydrogen, C_1-6 alkyl, a protecting group, or has a structure according to Formula II: , wherein:

Q is NH, O, or S; n is an integer from 0 to 4; R⁹ is hydrogen or C_1-3 alkyl; R¹⁰ and R¹¹ are each selected independently for each occurrence from the group consisting of hydrogen, alkyl, halogen, and –OH; and R¹² is H, C_1-3 alkyl, or –C(=NH)NR^12aR^12b, wherein R^12a and R^12b are each independently selected from the group consisting of hydrogen and C_1-3 alkyl; or R⁹ and R¹², together with the atoms to which they are attached, form a heterocycloalkyl ring system comprising at least one N. R¹³ and R¹⁴ are each independently selected from –OR⁶ and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of –OR⁶, N(R⁶)₂, and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, -OR⁶, -N(R⁶)₂, -NO₂, and –CN. [00216] In some embodiments the compound of Formula I having an ABCD ring system has a structure according to Formula Ib: AttyDktNo. R1119431100WO , wherein: R¹ is hydrogen or

R⁷ is NH₂ or OH; R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of -OH, -NH₂, and C_1-3 alkyl, said C_1-3 alkyl optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OH, -NH₂, -NO₂, and -CN. [00217] In some embodiments the compound of Formula I having an ABCD ring system has a structure according to Formula Ic: . [00218] In some

ring system has a structure according to Formula Id:

AttyDktNo. R1119431100WO . [00219] Preparation of of Formula Ia, Ib, Ic, and Id) disclosed herein follows

1, a Ring A compound and a Ring B compound are joined to form a Ring AB compound, and the Ring AB compound is joined via a glycosidic linkage with a Ring CD compound, each as described herein above. [00220] As a non-limiting example, the anomeric hydroxyl group of a Ring CD compound is converted to a suitable leaving group capable of reacting with a hydroxyl group on a Ring AB compound to form an interglycosidic linkage and allowed to react with the free hydroxyl group on the Ring AB 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, acetate, phosphate, O-pentenyl groups, mesylate, tosylate, and the like. In one embodiment, the Ring CD compound is treated with an activating agent such as trichloroacetonitrile to form an imidate, and the AB-Ring compound is added, followed by 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. Introduction of Substituent R^c [00221] In any of the foregoing embodiments, the compound of Formula I bears a substituent R^c. In some embodiments, R^c is H. In some embodiments, R^c C_1-6 alkyl. In some embodiments, R^c has a structure according to Formula II: , wherein:

Q is NH, O, or S; n is an integer from 0 to 4; R⁹ is hydrogen or C_1-3 alkyl; R¹⁰ and R¹¹ are each selected independently for each occurrence from the group consisting of hydrogen, alkyl, halogen, and –OH; and R¹² is H, C_1-3 alkyl, or –C(=NH)NR^12aR^12b, wherein R^12a and R^12b are each independently selected from the group consisting of hydrogen and C_1-3 alkyl; or AttyDktNo. R1119431100WO R⁹ and R¹², together with the atoms to which they are attached, form a heterocycloalkyl ring system comprising at least one N. [00222] Accordingly, in some embodiments, preparation of compounds of Formula I (e.g., compounds of Formula Ia, Ib, Ic, or Id) comprises reacting a suitable precursor of R^c with an appropriate ABCD ring compound to provide an ABCD ring compound having a protected substituent (R^d), and after additional appropriate steps, an ABCD ring compound having a substituent R^c. [00223] Referring to the structure of Formula (II), in some embodiments, Q is NH; R⁹ and R¹² are hydrogen; R¹⁰ and R¹¹ are each selected independently for each occurrence from the group consisting of hydrogen and F; and n is 2. In such embodiments, protected substituent R^d has a structure according to Formula (IIa): ; wherein:

Pg is a protecting group; Z is an amine precursor or a protected amino group; and the wavy line indicates a point of attachment to the amine of the B-ring. [00224] In some embodiments, R¹⁰, R^10a, R¹¹, and R^11a are hydrogen. [00225] In some embodiments, R¹⁰ is F, and R^10a, R¹¹, and R^11a are each hydrogen. [00226] In some embodiments, Z is azide (N₃). In some embodiments, Z is NHCBz. [00227] In some embodiments, Pg is benzyl. In some embodiments, Pg is para-methoxybenzyl (PMB). [00228] In some embodiments, R^c is selected from the group consisting of: R^d is one of:

AttyDktNo. R1119431100WO . [00229] In some

R^d is one of:

. [00230] In the foregoing

suitable reactant for introduction of the substituent R^d, such as the corresponding carboxylic acid or an activated version thereof (i.e., wherein the wavy line in the compound of Formula (IIa) denotes an OH group or leaving group). [00231] As described herein above, in some embodiments, the protected substituent R^d has a structure according to Formula (IIa), wherein R¹⁰, R^10a, R¹¹, and R^11a are each hydrogen, Z is NHCBz, and Pg is benzyl. In particular embodiments, R^d is ,

AttyDktNo. R1119431100WO and the corresponding reactant is (S)-2-(benzyloxy)-4-((2-oxo-2-phenyl-1 ^-ethyl)amino)butanoic acid, having the structure: . [00232] In some embodiments, the structure according to Formula (IIa), wherein R¹⁰ is F, and R^10a, R¹¹, and R^11a are each

, and Pg is para-methoxybenzyl. In particular embodiments, R^d is , and the corresponding reactant is (2R,3R)- 2-((4-methoxybenzyl)oxy)butanoic acid, having

the structure: . [00233] In one embodiment, the (2R,3R)- ((4-methoxybenzyl)oxy)butanoic acid may be

prepared from D-isoascorbic acid. For example, D-isoascorbic acid is protected as a ketal, and oxidatively cleaved to provide the ^-hydroxy ester. The ester is then reduced to the alcohol, and the primary hydroxyl group converted to an azide. The secondary hydroxyl group is then converted to a fluoro group with 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 benzylic ether (e.g., para-methoxybenzyl). Cleavage of the silyl ether and oxidation of the primary hydroxymethyl group to the corresponding carboxylic acid then provides the desired (2R,3R)-4-azido-3-fluoro-2-((4- methoxybenzyl)oxy)butanoic acid. One suitable synthesis of (2R,3R)-4-azido-3-fluoro-2-((4- methoxybenzyl)oxy)butanoic acid from D-isoascorbic acid is disclosed in International Patent Application No. PCT/IB2023/052011, which is incorporated by reference herein in its entirety. [00234] In some embodiments, protected substituent R^d has a structure according to Formula (IIa), and the reactant has a structure according to Formula (IIb): ; wherein each of Pg, R¹⁰, R¹¹, and Z are as

respect to Formula (IIa). [00235] In such embodiments, the reactant of Formula (IIb) may be allowed to react with the ABCD ring compound under suitable coupling conditions to form an ABCD ring compound bearing a protected AttyDktNo. R1119431100WO substituent R^d, followed by final global deprotection (e.g., by hydrogenolysis) to provide an ABCD ring compound having substituent R^c (i.e., a compound of Formula I). One of skill will recognize that the global deprotection (e.g., by hydrogenolysis) will also remove any benzyl, Cbz, or PMB groups present, and reduce any azide group present to the corresponding amine, providing the corresponding substituent R^c of the final compound of Formula I. Pharmaceutical Formulations and Compositions [00236] The compounds described herein can be considered useful as pharmaceutical compositions for administration to a subject in need thereof. Pharmaceutical compositions can comprise at least the compounds or salts described herein and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersing agents, suspending agents, and/or thickening agents. Accordingly, in another aspect is provided a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt or solvate of either thereof and at least one pharmaceutically acceptable excipient. For avoidance of doubt, reference herein to compounds of Formula I (e.g., with respect to compositions and methods of treatment) is intended to include compounds of each of Formulae Ia, Ib, Ic, and Id. [00237] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers comprising excipients and auxiliaries. Formulation can be modified depending upon the route of administration chosen. Pharmaceutical compositions comprising a compound or salt as described herein can be manufactured, for example, by lyophilizing the compound or salt, mixing, dissolving, emulsifying, encapsulating or entrapping the compound or salts. The pharmaceutical compositions can also include the compounds or salts, described herein in a free-base form or pharmaceutically acceptable salt form. [00238] Pharmaceutical compositions described herein can comprise at least one active ingredient (e.g., a compound or salt thereof). The active ingredients can be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin microcapsules and poly-(methylmethacrylate) microcapsules, respectively), in colloidal drug-delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. [00239] Pharmaceutical compositions as described herein often further can comprise more than one active compound (e.g., a compound or salt and other agents) as necessary 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 comprise a chemotherapeutic agent, cytotoxic agent, cytokine, growth-inhibitory agent, anti-hormonal agent, anti-angiogenic agent, and/or cardioprotectant. Such molecules can be present in combination in amounts that are effective for the purpose intended. [00240] The compositions and formulations can be sterilized. Sterilization can be accomplished by sterile filtration. [00241] The compositions described herein can be formulated for administration as an injection. Non-limiting examples of formulations for injection can include a sterile suspension, solution, or emulsion in oily or AttyDktNo. R1119431100WO 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 which increase the viscosity of the suspension. The suspension can also contain suitable stabilizers. Injections can be formulated for bolus injection or continuous infusion. Alternatively, the compositions described herein can be lyophilized or in powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. [00242] For parenteral administration, the compounds or salts can be formulated in a unit dosage injectable form (e.g., use letter solution, suspension, emulsion) in association with a pharmaceutically acceptable parenteral vehicle. Such vehicles can be inherently non-toxic, and non-therapeutic. Vehicles 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 minor amounts of additives such as substances that enhance isotonicity and chemical stability (e.g., buffers and preservatives). [00243] Sustained-release preparations can also be prepared. Examples of sustained-release preparations can 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). [00244] Pharmaceutical formulations described herein can be prepared for storage by mixing a compound or salt with a pharmaceutically acceptable carrier, excipient, and/or a stabilizer. This formulation can be a lyophilized formulation or an aqueous solution. Acceptable carriers, excipients, and/or stabilizers can be nontoxic to recipients at the dosages and concentrations used. Acceptable carriers, excipients, and/or stabilizers can include buffers; antioxidants; preservatives; polypeptides; proteins; hydrophilic polymers; amino acids; carbohydrates; chelating agents; sugars; salt-forming counter-ions; metal complexes; and/or non- ionic surfactants. [00245] The compounds of Formula I can be administered in the "native" form or, if desired, in the form of salts, esters, amides, prodrugs, clathrates, derivatives, and the like, provided the salt, ester, amide, prodrug, clathrate, or derivative is pharmacologically suitable, e.g., effective in treatment of a pathology and/or various symptoms thereof, e.g., as described herein. Salts, esters, amides, clathrates, prodrugs and other derivatives of the compounds of Formula I can be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry and described, for example, by March (1992) Advanced Organic Chemistry; Reactions, Mechanisms and Structure, 4th Ed. N.Y. Wiley-Interscience, and as described above. [00246] For example, a pharmaceutically acceptable salt can be prepared for any of the compounds of Formula I, described herein having a functionality capable of forming a salt. A pharmaceutically acceptable salt is any salt that retains the activity of the parent compound and does not impart any deleterious or untoward effect on the subject to which it is administered and in the context in which it is administered. [00247] Methods of formulating pharmaceutically compounds of Formula I as salts, esters, amide, prodrugs, and the like are well known to those of skill in the art. For example, salts can be prepared from the free base using conventional methodology that typically involves reaction with a suitable acid. Generally, the base form AttyDktNo. R1119431100WO of the drug is dissolved in a polar organic solvent such as methanol or ethanol and the acid is added thereto. The resulting salt either precipitates or can be brought out of solution by addition of a less polar solvent. Suitable acids for preparing acid addition salts include, but are not limited to both organic acids, e.g., 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, e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. An acid addition salt can be reconverted to the free base by treatment with a suitable base. [00248] In various embodiments, the compounds of Formula I described herein are useful for parenteral administration, topical administration, oral administration, nasal administration (or otherwise inhaled), rectal administration, or local administration, such as by aerosol or transdermally, for prophylactic and/or therapeutic treatment of one or more of the pathologies/indications described herein (e.g., pathologies characterized by excess amyloid plaque formation and/or deposition or undesired amyloid or pre-amyloid processing). [00249] The compounds of Formula I described herein can also be combined with a pharmaceutically acceptable carrier (excipient) to form a pharmacological composition. Pharmaceutically acceptable carriers can contain one or more physiologically acceptable compound(s) that act, for example, to stabilize the composition or to increase or decrease the absorption of the active agent(s). [00250] Other physiologically acceptable compounds, particularly of use in the preparation of tablets, capsules, gel caps, and the like include, but are not limited to binders, diluent/fillers, disintegrants, lubricants, suspending agents, and the like. [00251] Other physiologically acceptable compounds include wetting agents, emulsifying agents, dispersing agents or preservatives that are particularly useful for preventing the growth or action of microorganisms. [00252] The pharmaceutical compositions can be administered in a variety of unit dosage forms depending upon the method of administration. Suitable unit dosage forms, include, but are not limited to powders, tablets, pills, capsules, lozenges, suppositories, patches, nasal sprays, injectables, implantable sustained-release formulations, mucoadherent films, topical varnishes, lipid complexes, etc. [00253] Pharmaceutical compositions comprising the compounds of Formula I can be manufactured by means of conventional mixing, dissolving, granulating, dragee-making, levigating, 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 processing of the active agent(s) into preparations that can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. [00254] In certain embodiments, the active agents described herein (e.g., compounds of Formula I) and compositions comprising said compounds) are formulated for oral administration. For oral administration, suitable formulations can be readily formulated by combining the active agent(s) with pharmaceutically acceptable carriers suitable for oral delivery well known in the art. Such carriers enable the active agent(s) AttyDktNo. R1119431100WO 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 a patient to be treated. [00255] For administration by inhalation, the active agent(s) are conveniently delivered in the form of an aerosol spray from pressurized packs or a nebulizer, with the use of a suitable propellant. In the case of a pressurized aerosol the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch. [00256] In various embodiments the active agent(s) can be formulated in rectal or vaginal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides. Methods of formulating active agents for rectal or vaginal delivery are well known to those of skill in the art (see, e.g., Allen (2007) Suppositories, Pharmaceutical Press). [00257] For topical administration the compounds of Formula I can be formulated as solutions, gels, ointments, creams, suspensions, and the like as are well-known in the art. [00258] In certain embodiments the compounds of Formula I described herein are formulated for systemic administration (e.g., as an injectable) in accordance with standard methods well known to those of skill 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 certain emulsion formulations. The solution(s) can contain formulatory agents such as suspending, stabilizing and/or dispersing agents. In certain embodiments the active agent(s) can be provided in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. For transmucosal administration, and/or for blood/brain barrier passage, penetrants appropriate to the barrier to be permeated can be used in the formulation. Such penetrants are generally known in the art. Injectable formulations and inhalable formulations are generally provided as a sterile or substantially sterile formulation. [00259] In addition to the formulations described previously, the active agent(s) may also be formulated as a depot preparation. Such long-acting formulations can be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. [00260] In certain embodiments the active agent(s) described herein can also be delivered through the skin using conventional transdermal drug delivery systems, i.e., transdermal "patches" wherein the active agent(s) are typically contained within a laminated structure that serves as a drug delivery device to be affixed to the skin. In such a structure, the drug composition is typically contained in a layer, or "reservoir," underlying an upper backing layer. It will be appreciated that the term "reservoir" in this context refers to a quantity of "active ingredient(s)" that is ultimately available for delivery to the surface of the skin. Thus, for example, the "reservoir" may include the active ingredient(s) in an adhesive on a backing layer of the patch, or in any of a AttyDktNo. R1119431100WO variety of different matrix formulations known to those of skill in the art. The patch may contain a single reservoir, or it may contain multiple reservoirs. [00261] In certain embodiments the compounds of Formula I as described herein are formulated in a nanoemulsion. Nanoemulsions include but are not limited to oil in water (O/W) nanoemulsions, and water in oil (W/O) nanoemulsions. Nanoemulsions can be defined as emulsions with mean droplet diameters ranging from about 20 to about 1000 nm. Usually, the average droplet size is between about 20 nm or 50 nm and about 500 nm. The terms sub-micron emulsion (SME) and mini-emulsion are used as synonyms. [00262] Nanoemulsion formulations and methods of making such are well known to those of skill in the art and described for example in U.S. Patent 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, 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, an d5,152,923 and in Fanun et al. (2009) Microemulsions: Properties and Applications (Surfactant Science), CRC Press, Boca Ratan Fl. [00263] In certain embodiments, one or more active agents described herein can be provided as a "concentrate", e.g., in a storage container (e.g., in a premeasured volume) ready for dilution, or in a soluble capsule ready for addition to a volume of water, alcohol, hydrogen peroxide, or other diluent. [00264] In certain embodiments the compounds of Formula I described herein can also be administered using medical devices known in the art. For example, in one embodiment, a pharmaceutical composition of the invention can be administered with a needleless hypodermic injection device, such as the devices disclosed in U.S. Pat. No.5,399,163; U.S. Pat. No.5,383,851; U.S. Pat. No.5,312,335; U.S. Pat. No.5,064,413; U.S. Pat. No.4,941,880; U.S. Pat. No. 4,790,824; or U.S. Pat. No.4,596,556. Examples of well-known implants and modules useful for such deliver include, but are not limited to U.S. Pat. No. 4,487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Pat. No. 4,486,194, which discloses a therapeutic device for administering medicaments through the skin; U.S. Pat. No.4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; U.S. Pat. No. 4,447,224, which discloses a variable flow implantable infusion apparatus for continuous drug delivery; U.S. Pat. No. 4,439,196, which discloses an osmotic drug delivery system having multi-chamber compartments; and U.S. Pat. No.4,475,196, which discloses an osmotic drug delivery system. In a specific embodiment a compound of Formula I may be administered using a drug-eluting stent, for example one corresponding to those described in WO2001/87263 and related publications or those described by Perin (Perin, E.C., 2005). Many other such implants, delivery systems, and modules are known to those skilled in the art. [00265] The dosage to be administered of a compound of Formula I as described herein will vary according to the particular compound, the disease involved, the subject, and the nature and severity of the disease and the physical condition of the subject, and the selected route of administration. The appropriate dosage can be readily determined by a person skilled in the art. [00266] In certain embodiments the compositions may contain from 0.1%, e.g. from 0.1-70%, or from 5-60%, or preferably from 10-30%, of one or more compounds of Formula I, depending on the method of administration. AttyDktNo. R1119431100WO [00267] It will be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of Formula I as described herein will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the age and condition of the particular subject being treated, and that a physician will ultimately determine appropriate dosages to be used. This dosage may be repeated as often as appropriate. If side effects develop the amount and/or frequency of the dosage can be altered or reduced, in accordance with normal clinical practice. [00268] For the purposes of administration, the compounds of the present disclosure may be administered as a raw chemical or may be formulated as pharmaceutical compositions. Pharmaceutical compositions of the present disclosure comprise one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, and a pharmaceutically acceptable carrier. The one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, are present in the composition in an amount that is effective to treat a particular disease or condition of interest – for example, in an amount sufficient to treat a bacterial infection, and generally with acceptable toxicity to the subject. In some embodiments, the one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, are present in the composition in an amount that is effective to treat a particular disease or condition of interest. The antibacterial activity of compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, 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. [00269] Administration of one or more compounds of the disclosure, such as compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, in pure form or in an appropriate pharmaceutical composition can be carried out via any of the accepted modes of administration of agents for serving similar utilities. In some embodiments, the one or more compounds of the disclosure administered are compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of the foregoing, in pure form or in an appropriate pharmaceutical composition. [00270] The pharmaceutical compositions of the disclosure can be prepared by combining one or more compounds of the disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, with an appropriate pharmaceutically acceptable carrier, and may be formulated into preparations in solid, semi solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. Typical routes of administering such pharmaceutical compositions may include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal. The term parenteral as used herein may include subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques. Pharmaceutical compositions of the disclosure can be formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a subject. Compositions that will be administered to a subject or patient take the form of one or more dosage units, where for example, a tablet may be a single dosage unit, and a container of one or more compounds of the disclosure, or AttyDktNo. R1119431100WO pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, in aerosol form may hold a plurality of dosage units. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). [00271] A pharmaceutical composition of the disclosure may be in the form of a solid or liquid. In one aspect, the carrier(s) are particulate, so that the compositions are, for example, in tablet or powder form. The carrier(s) may be liquid, with the compositions being, for example, an oral syrup, injectable liquid or an aerosol, which may be useful in, for example, inhalatory administration. [00272] When intended for oral administration, pharmaceutical compositions of the present disclosure typically are either solid or liquid form, where semi solid, semi liquid, suspension and gel forms may be included within the forms considered herein as either solid or liquid. [00273] As a solid composition for oral administration, the pharmaceutical compositions may be formulated into a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like form. Such a solid composition will typically contain one or more inert diluents or edible carriers. [00274] In some embodiments, pharmaceutical compositions of the disclosure may be enriched to provide predominantly one enantiomer of a compound described herein. An enantiomerically enriched mixture may comprise, for example, at least 60 mol percent of one enantiomer, 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. In some embodiments, the compositions described herein enriched in one enantiomer may be substantially free of the other enantiomer, wherein substantially free may mean that the substance in question makes up less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% as compared to the amount of the other enantiomer, e.g., in the pharmaceutical composition or compound mixture. For example, if a pharmaceutical composition or compound mixture contains 98 grams of a first enantiomer and 2 grams of a second enantiomer, it would be said to contain 98 mol percent of the first enantiomer and only 2 mol percent of the second enantiomer. [00275] In some embodiments, the pharmaceutical compositions of the disclosure may be enriched to provide predominantly one diastereomer of a compound disclosed herein. A diastereomerically enriched mixture may comprise, for example, at least 60 mol percent of one diastereomer, 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. In some embodiments, the compositions described herein enriched in one diastereomer may be substantially free of other diastereomers, wherein substantially free may mean that the substance in question makes up less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% as compared to the amount of other diastereomers, e.g., in the pharmaceutical composition or compound mixture. [00276] Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods well known to those skilled in the art, such as, for example, by chromatography and/or fractional crystallization. Enantiomers may be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound AttyDktNo. R1119431100WO (e.g., chiral auxiliary such as a chiral alcohol or Mosher’s acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Enantiomers can also be separated by use of a chiral HPLC column. Also, some of the compounds of the disclosure may be atropisomers or rotameric forms and are considered as part of this disclosure. Therapeutic Applications [00277] The compounds, compositions, and methods of the present disclosure can be useful for a plurality of different subjects including, but not limited to, a mammal, a human, a non-human mammal, a domesticated animal (e.g., laboratory animals, household pets, or livestock), a non-domesticated animal (e.g., wildlife), a dog, a cat, a rodent, a mouse, a hamster, a cow, a bird, a chicken, a fish, a pig, a horse, a goat, a sheep, a rabbit, or any combination thereof. [00278] The compounds and compositions described herein can be useful as therapeutics, for example, a treatment that can be administered to a subject in need thereof. A therapeutic effect of the present disclosure can be obtained in a subject by reduction, suppression, remission, or eradication of a disease state, including, but not limited to, a symptom thereof. A therapeutic effect in a subject having a disease or condition, or pre- disposed to have or is beginning to have the disease or condition, can be obtained by a reduction, a suppression, a prevention, a remission, or an eradication of the condition or disease, or pre-condition or pre-disease state. [00279] In practicing the methods described herein, therapeutically effective amounts of the compositions described herein can be administered to a subject in need thereof, often for treating and/or preventing a condition or progression thereof. A pharmaceutical composition can affect the physiology of the subject, such as the immune system, an inflammatory response, or other physiologic affect. A therapeutically effective amount can vary depending on the severity of the disease, the age and relative health of the subject, the potency of the compounds used, and other factors. [00280] Treat and/or treating can refer to any indicia of success in the treatment or amelioration of the disease or condition. Treating can include, for example, reducing, delaying or alleviating the severity of one or more symptoms of the disease or condition, or it can include reducing the frequency with which symptoms of a disease, defect, disorder, or adverse condition, and the like, are experienced by a patient. Treat can be used herein to refer to a method that results in some level of treatment or amelioration of the disease or condition and can contemplate a range of results directed to that end, including but not restricted to prevention of the condition entirely. [00281] Prevent, preventing and the like can refer to the prevention of the disease or condition, e.g., tumor formation, in the 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 later develop the tumor or other form of cancer, then the disease has been prevented, at least over a period of time, in that individual. Preventing can also refer to preventing re-occurrence of a disease or condition in a patient that has previously been treated for the disease or condition, e.g., by preventing relapse. AttyDktNo. R1119431100WO [00282] A therapeutically effective amount can be the amount of a composition or an active component thereof sufficient to provide a beneficial effect or to otherwise reduce a detrimental non-beneficial event to the individual to whom the composition is administered. A therapeutically effective dose can be a dose that produces one or more desired or desirable (e.g., beneficial) effects for which it is administered, such administration occurring one or more times over a given period of time. An exact dose can depend on the purpose of the treatment and can be ascertainable by one skilled in the art using known techniques. Methods of Treating Bacterial Infection [00283] This disclosure features methods of and uses for treating a bacterial infection comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of the 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 disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof. [00284] Compounds, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and pharmaceutical compositions of the present disclosure (e.g., compounds of Formula I) may possess antibacterial activity against a wide spectrum of gram-positive and gram-negative bacteria. Representative susceptible organisms may include those gram-positive, gram-negative, aerobic, and facultative anaerobic organisms whose growth can be inhibited by the compounds of the 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. [00285] Accordingly, compounds, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and pharmaceutical compositions comprising the compounds or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, as disclosed herein can be used, for example, for treating bacterial infections and for treating conditions caused by the bacterial infection. Bacterial infections susceptible to treatment according to the present disclosure may include primary infections and co-infections caused by a species of bacteria and one or more additional infectious agents such as, for example, bacteria, virus, parasite and fungi. [00286] In one aspect is provided a method of treating a bacterial infection, the method comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any thereof, or a pharmaceutical composition comprising one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any thereof. [00287] In some embodiments, the bacterial infection is with Gram-positive, Gram-negative, aerobic, or facultative anaerobic bacteria. AttyDktNo. R1119431100WO [00288] In some embodiments, the bacterial infection is with Gram-positive bacteria. Examples of Gram- positive bacteria include, but are not limited to, Staphylococcus sp., Lactobacillus sp., Streptococcus sp., Sarcina sp., Bacillus sp., Peptococcus sp., Corynebacterium sp., and Enterococcus sp. In some embodiments, the Gram-positive bacterial infection is an infection with a Staphylococcus sp., Lactobacillus sp., Streptococcus sp., Sarcina sp., Bacillus sp., Peptococcus sp., Corynebacterium sp., or Enterococcus sp. 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). [00289] In some embodiments, the bacterial infection is with Gram-negative bacteria. Examples of Gram- negative bacteria include, but are not limited to, Escherichia sp., Enterobacter sp., Klebsiella sp., Pseudomonas sp., Acinetobacter sp., Proteus sp., Campylobacter sp., Citrobacter sp., Neisseria sp., Salmonella sp., Shigella sp., Serratia sp., Haemophilus sp., Brucella sp., Francisella sp., Yersinia sp., Burkholderia sp., and Moraxella sp. [00290] In some embodiments, the bacterial infection is caused by 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; Franciscellaceae (e.g., Francisella tularensis); Neisseria species; and Enterobacteriaceae, such as Serratia, Proteus, Klebsiella, Enterobacter, Citrobacter, Salmonella, Providencia, Yersinia (e.g., Yersinia pestis), Morganella, Cedecea, Edwardsiella species, Acinetobacter, and Escherichia coli. [00291] In some embodiments, the Gram-negative bacterial infection is an infection with an Escherichia sp., Enterobacter sp., Klebsiella sp., Pseudomonas sp., Acinetobacter sp., Proteus sp., Campylobacter sp., Citrobacter sp., Neisseria sp., Salmonella sp., Shigella sp., Serratia sp., Haemophilus sp., Brucella sp., Francisella sp., Yersinia sp., Burkholderia sp., or Moraxella sp. In some embodiments, the Gram-negative bacterial infection is an infection with an Enterobacteriaceae, Haemophilus, Franciscellaceae, or Neisseria species. In some embodiments, the bacterial infection is caused by an Enterobacteriaceae. In some embodiments, the bacterial infection is caused by an extended spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae. [00292] In some embodiments, the Gram-negative bacterial infection is an infection with 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 with Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxidans, [00293] In some embodiments, the bacterial infection is an infection with a member of the order of Enterobacterales. AttyDktNo. R1119431100WO [00294] In some embodiments, the bacterial infection is an infection with an Escherichia spp., a Klebsiella spp., a Proteus spp., a Citrobacter spp., a Morganella spp., a Providencia spp., a Yersinia spp., an Enterobacter spp., a Salmonella spp., or a Serratia spp. [00295] In some embodiments, the bacterial infection is an infection with a Moraxella spp., a Pseudomonas spp., an Acinetobacter spp., a Mycobacterium spp., a Staphylococcus spp., a Bacillus spp., a Francisella spp., or a Burkholderia spp. [00296] In some embodiments, the bacterial infection is an infection with a 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., Peptococcus sp., Salmonella sp., Shigella sp., Serratia sp., Haemophilus sp., Brucella sp., Francisella sp., Yersinia sp., Corynebacterium sp., Moraxella sp., or Burkholderia sp., or Enterococcus sp. [00297] In some embodiments, the bacterial infection is caused by multidrug resistant (MDR) bacteria. In some embodiments, the bacterial infection is caused by a multidrug-resistant biothreat pathogen, including those defined as such by the United States Centers for Disease Control and Prevention’s Category A/B/C bioterrorism agent designation. In certain such embodiments, the biothreat pathogen is Gram-negative bacteria. [00298] In some embodiments, one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, or pharmaceutical compositions of the disclosure may be useful in treating diseases or conditions caused by a bacterial infection as disclosed herein, including, but not limited to, skin infections; pneumonia; nosocomial pneumonia; sepsis; respiratory tract infections (e.g., lower respiratory tract or upper respiratory tract infections); lung infection in cystic fibrosis patients; acute exacerbation of chronic bronchitis; community acquired pneumonia; nosocomial pneumonia (including ventilator-associated pneumonia (VAP)); hospital or community acquired infections caused by Gram-negative bacteria; diseases of the upper airways; diffuse panbronchiolitis; tonsillitis; pharyngitis; acute sinusitis and otitis including mastoiditis; urinary tract and genital infections, for example, complicated urinary tract infections, cystitis, resistant or relapsing cystitis, urethritis, pyelonephritis, acute pyelonephritis, endometritis, prostatitis, salpingitis, and epididymitis; ocular infections such as conjunctivitis, corneal ulcer, iridocyclitis and post-operative infection in radial keratotomy surgery patients; blood infections, for example septicemia or bacteremia; infections of the skin and soft tissues, for example infective dermatitis, infected wounds, infected burns, phlegmon, folliculitis and impetigo; bone and joint infections such as osteomyelitis and septic arthritis; gastrointestinal infections, for example dysentery, enteritis, colitis, necrotizing enterocolitis and anorectal infections; intraabdominal infections, including, but not limited to, complicated intraabdominal infections, typhoid fever, infectious diarrhea, peritonitis with appendicitis, pelviperitonitis, and intra- abdominal abscesses; infections in the oral region, for example infections after dental operations; other infections, for example, melioidosis, infectious endocarditis, sexually transmitted diseases caused by Gram- negative or Gram-positive bacteria, hepatic abscesses, cholecystitis, cholangitis, mastitis as well as meningitis, and infections of the nervous systems. AttyDktNo. R1119431100WO [00299] In some embodiments, the method disclosed herein treats infections selected from the group consisting of complicated intraabdominal infections, complicated urinary tract infections, resistant or relapsing cystitis, acute pyelonephritis, and bacteremia. [00300] In a further aspect, the disclosure provides for use of one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, to treat a bacterial infection as described herein in a subject in need thereof. [00301] In still a further aspect, provided herein is one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, for use in a method of treating a bacterial infection in a subject in need thereof. [00302] In another aspect, the disclosure provides for one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of the foregoing, for use as a medicament. [00303] In yet a further aspect, provided herein is the use of one or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, in the manufacture of a medicament for treating a bacterial infection in a subject in need thereof. [00304] In still a further aspect, provided herein is one or more pharmaceutical compositions of the disclosure for use in a method of treating a bacterial infection in a subject in need thereof. [00305] In a still further aspect, provided herein is the use of one or more pharmaceutical compositions of the disclosure in the manufacture of a medicament for treating a bacterial infection in a subject in need thereof. [00306] In any of the foregoing methods and uses as disclosed herein, the subject may be a mammal. In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human. Administration [00307] One or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, or compositions of any thereof, may be administered in a therapeutically effective amount, which will vary depending upon a variety of factors including: the activity of the specific compound employed; the metabolic stability and length of action of the compound; the age, body 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 undergoing therapy. [00308] Administration of one or more compounds of the disclosure, such as compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, in pure form or in an appropriate pharmaceutical composition can be carried out via any of the accepted modes of administration of agents for serving similar utilities. In some embodiments, the one or more compounds of the disclosure administered are compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of the foregoing, in pure form or in an appropriate pharmaceutical composition. [00309] Pharmaceutical compositions can be used in the methods described herein and can be administered to a subject in need thereof using a technique known to one of ordinary skill in the art which can be suitable as a AttyDktNo. R1119431100WO therapy for the disease or condition affecting the subject. One of ordinary skill in the art would understand that the amount, duration and frequency of administration of a pharmaceutical composition described herein to a subject in need thereof depends on several factors including, for example but not limited to, the health of the subject, the specific disease or condition of the patient, the grade or level of a specific disease or condition of the patient, the additional therapeutics the subject is being or has been administered, and the like. [00310] The methods and compositions described herein can be for administration to a subject in need thereof. Often, administration of the compositions described herein can include routes of administration, non-limiting examples of administration routes include oral, intravenous, intraarterial, subcutaneous, subdural, intramuscular, intracranial, intrasternal, intratumoral, or intraperitoneally. Additionally, a pharmaceutical composition can be administered to a subject by additional routes of administration, for example, by inhalation, dermal, intranasal, or intrathecal administration. [00311] Compositions of the present disclosure can be administered to a subject in need thereof in a first administration, and in one or more additional administrations. The one or more additional administrations can be administered to the subject in need thereof minutes, hours, days, weeks or months following the first administration. Any one of the additional administrations can be administered to the subject in need thereof less than 21 days, or less than 14 days, less than 10 days, less than 7 days, less than 4 days or less than 1 day after the first administration. The one or more administrations can occur more than once per day, more than once per week or more than once per month. The administrations can be weekly, biweekly (every two weeks), every three weeks, monthly or bimonthly. [00312] One or more compounds of Formula I, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers of any of these, may also be administered simultaneously with, prior to, or after administration of one or more other therapeutic agents. Such combination therapy may include administration of a single pharmaceutical dosage formulation that contains one or more compounds of the disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and one or more additional active agents, as well as administration of one or more compounds of the disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and each active agent in its own separate pharmaceutical dosage formulation. For example, one or more compounds of the disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and the other active agent can be administered to the subject together in a single oral dosage composition such as a tablet or capsule, or each agent administered in separate oral dosage formulations. Where separate dosage formulations are used, the one or more compounds of the disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and one or more additional active agents can be administered at essentially the same time, e.g., concurrently, or at separately staggered times, e.g., sequentially; combination therapy is understood to include all these regimens. AttyDktNo. R1119431100WO Kits [00313] In some aspects, the present disclosure provides a kit comprising a compound as disclosed herein, or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof, or a composition comprising said compound or pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof, and instructions. [00314] In some embodiments, the present disclosure provides a kit comprising a pharmaceutical composition as disclosed herein and instructions. [00315] In some embodiments, the instructions comprise instructions for administering the compound, salt or pharmaceutical composition as disclosed to a subject in need thereof. In some embodiments, the kit comprises a compound, salt or pharmaceutical composition disclosed herein, packaged in a low moisture vapor transmission container with a desiccant. Optionally, a label is on or associated with the container. In addition, a label may be used to indicate that the contents are to be used for a specific therapeutic application. In some embodiments, the label includes directions for use of the contents, such as in the methods described herein. In some embodiments, a compound, salt or pharmaceutical composition disclosed herein is presented in a pack or container that contains one or more unit dosage forms comprising the compound, salt or pharmaceutical composition disclosed herein. The pack may contain metal or plastic foil, such as a blister pack. [00316] A "kit" as used herein may include a container for containing at least one pharmaceutical composition or compound of the disclosure and may also include divided containers such as a divided bottle or a divided foil packet. The container can be in any conventional shape or form as known in the art that is made of a pharmaceutically acceptable material, for example a paper or cardboard box, a glass or plastic bottle or jar, a resealable bag (for example, to hold a "refill" of tablets for placement into a different container), or a blister pack with individual doses for pressing out of the pack according to a therapeutic schedule. [00317] The kits of the present disclosure may also include, in addition to one or more compounds or pharmaceutical compositions of the present disclosure, one or more additional pharmaceutically active compounds or pharmaceutical compositions. For example, the additional compound may be a second antibacterial compound or the additional pharmaceutical composition may comprise a second antibacterial compound. The additional compounds or pharmaceutical compositions 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. Likewise, the additional compounds or pharmaceutical compositions can be administered at the same time as the one or more compounds or pharmaceutical compositions of the present disclosure or at different times. [00318] Although the technology herein has been described with reference to particular embodiments, it is to be understood that these embodiments are merely illustrative of the principles and applications of the present technology. It will be apparent to those skilled in the art that various modifications and variations can be made to the method and apparatus of the present technology without departing from the spirit and scope of the technology. Thus, it is intended that the present technology include modifications and variations that are AttyDktNo. R1119431100WO within the scope of the appended claims and their equivalents. Accordingly, the disclosure is not limited except as by the appended claims. [00319] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. Any ranges cited herein are inclusive. [00320] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, 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. [00321] 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. [00322] Aspects of the present technology are more fully illustrated with reference to the following examples. Before describing several exemplary embodiments of the technology, it is to be understood that the technology is not limited to the details of construction or process steps set forth in the following description. The technology is capable of other embodiments and of being practiced or being carried out in various ways. The following examples are set forth to illustrate certain aspects of the present technology and are not to be construed as limiting thereof. It is understood that one skilled in the art may be able to make these compounds by similar methods or by combining other methods known to one skilled in the art. It is also understood that one skilled in the art would be able to make, in a similar manner as described below, further compounds within the scope of the present disclosure by using appropriate starting materials and modifying the synthetic route as needed. In general, starting materials and reagents can be obtained from commercial vendors or synthesized according to sources known to those skilled in the art or prepared as described herein. [00323] Reagent/reactant names given are as named 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 include more than one molar equivalent of the acid. EXEMPLIFICATION AttyDktNo. R1119431100WO ABBREVIATIONS 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 Doublet DAST Diaminosulfur trifluoride DCM Dichloromethane (methylene chloride) dd Doublet of doublets DIAD Diisopropyl azodicarboxylate DMAP N,N-Dimethylaminopyridine DMEM Dulbecco’s Modified Eagle 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(s) EA or EtOAc Ethyl acetate Et₂O Diethyl ether EtOH Ethanol g Gram(s) h Hour(s) HPLC High-pressure liquid chromatography LAH Lithium aluminum hydride LC/MS Liquid chromatography/mass spectrometry LiOH Lithium hydroxide m Multiplet AttyDktNo. R1119431100WO M Molar MeCN Acetonitrile MeOH Methyl alcohol min Minute(s) mmol Millimole MPLC Medium pressure liquid chromatography MS Mass spectrometry n- Normal (nonbranched) 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 PE Petroleum ether 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 Reverse-phase high-pressure liquid chromatography R_t Retention time rt Room temperature s Singlet t Triplet t- Tertiary t-BuOH t-butanol TBAI Tetra-n-Butylammonium iodide TBSCl t-Butyldimethylsilyl chloride TEA Triethylamine TfN₃ Trifluoromethanesulfonyl azide TFA Trifluoroacetic acid AttyDktNo. R1119431100WO THF Tetrahydrofuran TMS Trimethylsilyl TMSOTf Trimethylsilyl trifluormethanesulfonate TolSH p-Thiocresol TsCl para-toluenesulfonyl chloride USP United States Pharmacopeia wt% Weight percent GENERAL [00324] All temperatures are in degrees Celsius (°C) and are uncorrected. Reagent grade chemicals and anhydrous solvent were purchased from commercial sources and unless otherwise mentioned, were used without further purification. The names of the products were determined using the naming software included in the BIOVIA Notebook. Flash chromatography and reverse phase chromatography was performed on Teledyne Isco instruments using pre-packaged disposable SiO₂ or C-18 stationary phase columns with eluent flow rate range of 15 to 200 mL/min, UV detection. The analytical HPLC chromatograms were performed using an Agilent 1100 series instrument with DAD detector (190 nm to 300 nm). The mass spectra were recorded with a Waters Micromass ZQ detector at 130 ºC. The mass spectrometer was equipped with an electrospray ion source (ESI) operated in a positive ion mode and was set to scan between m/z 150-750 with a scan time of 0.2 s. Intermediates were analyzed by HPLC/MS on a Kinetex EVO C18 (5 uM, 4.6 x 50 mm) using a low pH buffer gradient of 5% to 100% of MeCN in H₂O (0.1% HCOOH) over 1.75 min at 2.2 mL/min for a 3.5 min run. The mass spectra were recorded with a Waters ACQUITY SQD (source; 150 ºC, desolvation gas; 900 L/h, cone; 50). The mass spectrometer was equipped with an electrospray ion source (ESI) operated in a positive ion mode and was set to scan between m/z 150-2000 with a scan time of 0.2 s. Intermediates were analyzed by UPLC/MS on a Waters CSH C18 (1.7 uM, 4.6 x 50 mm) using a low pH buffer gradient of 5% to 100% of MeCN in H₂O (10 mM ammonium formate) over 2.2 min at 0.7 mL/min for a 3 min run. The ¹H NMR spectra were recorded on a Bruker UltraShield 500 MHz/54 mm instrument (BZH 43/500/70B, D221/54-3209) and chemical shifts are quoted in parts per million (ppm). Example 1. Synthesis of (3R,5S,6R)-3-azido-6-(azidomethyl)-5-benzyloxy-tetrahydropyran-2-ol (Ring A1) Step A. Synthesis of 2-[

oxan-3-yl]isoindole-1,3- dione AttyDktNo. R1119431100WO [00325] To a solution of triol (D-glucosamine; 45 g, 208.72 mmol, 1.0 eq) in

anhydride (30.9 g, 208.72 mmol, 1.0 eq) and NaHCO₃ (35.1 g, 417.45 mmol, 2.0 eq). The resulting mixture was stirred at 25℃ for 2h. Then the reaction mixture was concentrated under vacuum to give the title compound (116 g) as a yellow solid which was directly consumed in the next step without further purification. LCMS [M+1]⁺: 310.3. Step B. Synthesis of [(2R,3S,4R,5R)-3,4,6-triacetyloxy-5-(1,3-dioxoisoindol-2-yl)oxan-2-yl]methyl acetate [00326] To a solution oxan-3-yl]isoindole-

1,3-dione (116 g) in was . resulting mixture was stirred at 50 ^oC for 3 h. The solvent was then concentrated under vacuum and the resulting brown oil was diluted with CH₂Cl₂ (800 mL). The resulting mixture was washed with water (500 mL). The organic layer was concentrated under vacuum to give the title compound (103 g) as a yellow oil which was directly consumed in next step without further purification. LCMS [M+18]⁺: 495.1.¹H NMR (400 MHz, CDCl₃) δ 7.88-7.86 (m, 2 H), 7.77-7.76 (m, 2 H), 6.46-6.44 (d, 1 H), 5.96-5.92 (m, 1 H), 5.53-5.52 (d, 1 H), 4.69-4.65 (m, 1 H), 4.23- 4.17 (m, 3H) , 2.21 (s, 3 H), 2.07 (s, 3 H), 2.01 (s, 3 H), 1.86 (s, 3 H). Step C. Synthesis of [(2R,3S,4R,5R)-3,4-diacetyloxy-5-(1,3-dioxoisoindol-2-yl)-6-(4- methylphenyl)sulfanyloxan-2-yl]methyl acetate [00327] To a

2-yl)oxan-2- yl]methyl acetate (103 g, 218 mmol) in DCM (1000 mL) was added p-thiocresol (32 g, 290 mmol, 1.3 eq.). This was followed by the dropwise addition of BF₃ ^OEt₂ (160 ml, 6.0 eq) over 10 mins. After stirring at room temperature for 24 h, the reaction mixture was quenched with saturated NaHCO₃ aqueous solution. The resulting mixture was extracted with DCM. The organic layers were combined and concentrated under AttyDktNo. R1119431100WO vacuum. The residue was purified via silica gel chromatography column with PE/EA (2/1) to give the title compound (93 g, 78% from (3R,4R,5S,6R)-3-amino-6-(hydroxymethyl)oxane-2,4,5-triol (D-glucosamine)) as a yellow solid. LCMS [M+18]⁺: 559.2.¹H NMR (400 MHz, CDCl₃) δ 7.89-7.85 (m, 2 H), 7.77-7.74 (m, 2 H), 7.32-7.06 (dd, 4 H), 5.82-5.79 (m, 1 H), 5.64-5.62 (d, 1 H), 5.49-5.48 (d, 1 H), 4.65-4.59 (t, 1H) , 4.23- 4.08 (m, 3 H), 2.31 (s, 3 H), 2.18 (s, 3 H), 2.05 (s, 3 H), 1.84 (s, 3 H). Step D. Synthesis of 2-[(3R,4R,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-(4- methylphenyl)sulfanyloxan-3-yl]isoindole-1,3-dione [00328] To a 2-yl)-6-(4-

methylphenyl) g, was added NaOMe (1.8 g, 34.3 mmol, 0.2 eq.). After stirring at room temperature for 2 h, the reaction mixture was concentrated under vacuum. The residue was purified via silica gel chromatography column with PE/EA (1/2) to give the title compound (48 g, 67%) as a yellow solid. LCMS [M+18]⁺: 433.4.¹H NMR (400 MHz, DMSO-d₆) δ 7.86-7.82 (m, 2 H), 7.75-7.72 (m, 2 H), 7.27-7.26 (m, 2 H), 7.06-7.04 (d, 2 H), 5.58-5.55 (d, 1 H), 4.46-4.36 (m, 2 H), 4.15-4.12 (m, 1H), 3.96-3.94 (m, 2 H), 3.71-3.69 (m, 1 H), 3.61-3.60 (m, 1 H), 2.94-2.92 (m, 1 H), 2.56 (m, 1 H), 2.29 (s, 3 H). Step E. Synthesis of 2-[(4aR,7R,8R,8aS)-8-hydroxy-6-(4-methylphenyl)sulfanyl-2-phenyl-4,4a,6,7,8,8a- hexahydropyrano[3,2-d][1,3]dioxin-7-yl]isoindole-1,3-dione

[00329] To a solution of 2-[(3R,4R,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-(4- methylphenyl)sulfanyloxan-3-yl]isoindole-1,3-dione (48 g, 115.6 mmol, 1.0 eq) in DMF (600 mL) were added PTSA (5.8 g, 34.6 mmol, 0.3 eq.) and benzaldehyde dimethyl acetal (45 mL, 346 mmol, 3 eq.). After stirring at 75 ^oC for 16 h, the reaction was quenched with saturated NaHCO₃ aqueous solution. The resulting mixture was extracted with DCM and the organic layers were concentrated under vacuum. The residue was purified via silica gel chromatography column with petroleum ether /acetate (3/1) to give the title compound (50 g, 86% yield) as a colorless solid. LCMS [M+18]⁺: 521.3.¹H NMR (400 MHz, DMSO-d₆) δ 7.96-7.93 (m, 4 H), AttyDktNo. R1119431100WO 7.53-7.47 (m, 5 H), 7.35-7.12 (dd, 4 H), 5.64-5.61 (m, 3 H), 4.53-4.36 (m, 3 H), 4.22-4.07 (m, 2 H), 3.87 (s, 1H), 2.31 (s, 3 H). Step F. Synthesis of 2-[(4aR,7R,8R,8aS)-6-(4-methylphenyl)sulfanyl-8-phenoxycarbothioyloxy-2- phenyl-4,4a,6,7,8,8a-hexahydropyrano[3,2-d][1,3]dioxin-7-yl]isoindole-1,3-dione [00330] To a phenyl-4,4a,6,7,8,8a-

hexahydropyrano g, (800 mL) was added DMAP (36.5 g, 297 mmol, 3 eq.) and phenyl chlorothionocarbonate (20 mL, 148.54 mmol, 1.5 eq.). After stirring at room temperature for 16 h, the reaction mixture was concentrated under vacuum and the residue was purified via silica gel chromatography column with PE/EA (3/1) to give the title compound (53 g, 84%) as a colorless solid. LCMS [M+18]⁺: 657.1.¹H NMR (400 MHz, DMSO-d₆) δ 7.92-7.76 (m, 4 H), 7.49-7.39 (m, 6 H), 7.25-7.18 (m, 4 H), 7.04-7.03 (d, 2 H), 6.84-6.78 (m, 2 H), 6.27-6.24 (dd, 1 H), 5.76-5.73 (d, 1H) , 5.60 (s, 1 H), 4.95-4.90 (t, 1 H), 4.78-4.77 (d, 1 H), 4.46-4.42 (dd, 1 H), 4.13-4.10 (dd, 1 H), 3.80 (s, 1 H), 2.32 (s, 1 H), 2.32 (s, 3 H). Step G. Synthesis of 2-[(4aR,7R,8aS)-6-(4-methylphenyl)sulfanyl-2-phenyl-4,4a,6,7,8,8a- hexahydropyrano[3,2-d][1,3]dioxin-7-yl]isoindole-1,3-dione [00331] To a

2- phenyl-4,4a,6,7,8,8a-hexahydropyrano[3,2-d][1,3]dioxin-7-yl]isoindole-1,3-dione (53 g, 82.3 mmol) in toluene (700 mL) was added Bu₃SnH (72 g, 248.8 mmol, 3 eq.) and AIBN (6.7 g, 4.1 mmol, 0.05 eq.). After stirring at 115 ^oC for 1.5 h, the reaction mixture was concentrated under vacuum. The residue was purified via silica gel chromatography column with PE/EA (3/1) to give the title compound (19 g, 48%) as a colorless solid. LCMS [M+18]⁺: 505.2.¹H NMR (400 MHz, DMSO-d₆) δ 7.84-7.71 (m, 4 H), 7.52-7.45 (m, 2 H), 7.41- AttyDktNo. R1119431100WO 7.36 (m, 5 H), 7.06-7.04 (d, 2 H), 4.64-5.61(d, 1 H), 5.54 (s, 1 H), 4.73-4.66 (m, 1H) , 4.39-4.36 (d, 1 H), 4.16 (s, 1 H), 4.09-4.06 (dd, 1 H), 3.86-3.84 (d, 1 H), 2.81-2.74 (m, 1 H), 2.32 (s, 3 H), 2.23-2.18 (m, 1 H). Step H. Synthesis of (4aR,7R,8aS)-6-(4-methylphenyl)sulfanyl-2-phenyl-4,4a,6,7,8,8a- hexahydropyrano[3,2-d][1,3]dioxin-7-amine [00332] To a 2-phenyl-4,4a,6,7,8,8a- hexahydropyrano

t-BuOH (500 mL) was added ethylenediamine (14 g, 223.7 mmol, 6.0 eq). After stirring at 95 ^oC for 16 h, the reaction mixture was concentrated under vacuum. The residue was purified via silica gel chromatography column with PE/EA (1/3) to give the title compound (12 g, 85%) as a colorless solid. LCMS [M+1]⁺: 358.2.¹H NMR (400 MHz, DMSO- d₆) δ 7.56-7.54 (d, 2 H), 7.46-7.42 (m, 5 H), 7.18-7.16 (d, 2 H), 5.60 (s, 1 H), 4.54-4.52(d, 1 H), 4.14-4.04 (m, 3 H), 3.59 (s, 1H) , 2.84-2.78 (m, 1 H), 2.35 (s, 3 H), 2.10-2.06 (m, 1 H), 1.67-1.55 (m, 3 H). Step I. Synthesis of (4aR,7R,8aS)-7-azido-6-(4-methylphenyl)sulfanyl-2-phenyl-4,4a,6,7,8,8a- hexahydropyrano[3,2-d][1,3]dioxine [00333] Sodium azide

cooled to 0 °C. Triflic anhydride (28.4 g, 96 mmol, 3 eq.) was dissolved in 40 mL of DCM and added to the NaN₃ solution. The reaction was stirred at 0 ^oC for 2 h. (4aR,7R,8aS)-6-(4-methylphenyl)sulfanyl-2-phenyl-4,4a,6,7,8,8a- hexahydropyrano[3,2-d][1,3]dioxin-7-amine (12 g, 32 mmol) was dissolved in 80 mL of water and CuSO₄ (50 mg, 3.2 mmol, 0.01 eq.) was added to form a malachite-colored solution. The TfN₃ solution in DCM (120 mL) was then added to this solution. After a slow addition of MeOH (40 mL) over 2 minutes the reaction was stirred at room temperature for 2 days. Then the reaction was quenched with saturated NaHCO3 aqueous solution. The resulting mixture was extracted with DCM and the organic layers were concentrated under vacuum. The residue was purified via silica gel chromatography column with PE/EA (5/1) to give the title compound (9 g, 70%) as a colorless solid. LCMS [M+18]⁺: 401.1. ¹H NMR (400 MHz, DMSO-d₆) δ 7.53- 7.51 (d, 2 H), 7.42-7.38 (m, 5 H), 7.15-7.13 (d, 2 H), 5.58 (s, 1 H), 4.81-4.79(d, 2 H), 4.14-4.02 (m, 3 H), 2.31 (s, 3H), 2.27-2.22 (m, 1 H), 1.88-1.80 (m, 1 H). Step J. Synthesis of [(2R,3S,5R)-5-azido-6-(4-methylphenyl)sulfanyl-3-phenylmethoxyoxan-2- yl]methanol AttyDktNo. R1119431100WO [00334] To a 2-phenyl-4,4a,6,7,8,8a- hexahydropyrano[3,2-d]

a solution of BH₃ ^THF (1 M, 23.4 mL, 23.4 mmol, 1.0 eq.) and TMSOTf (0.5 g, 2.34 mmol, 0.1 eq.). After stirring at 0 °C to 3 °C temperature for 2 h, the reaction mixture was quenched with saturated NaHCO₃ aqueous solution. The resulting mixture was extracted with DCM and the organic layers were concentrated under vacuum. The residue was purified via silica gel chromatography column with PE/EA (5/1) to give the title compound (4.7 g, 53 %) as a colorless solid. LCMS [M+18]⁺: 403.4. ¹H NMR (400 MHz, DMSO-d₆) δ 7.42-7.40 (d, 2 H), 7.37-7.27 (m, 5 H), 7.14-7.20 (d, 2 H), 4.78-4.75 (s, 1 H), 4.74-4.72(d, 2 H), 4.65-4.40 (dd, 2 H), 3.70 (s, 1H) , 3.64-3.50 (m, 4 H), 3.34 (s, 1 H), 2.52-2.50 (m, 1 H), 2.28 (s, 3 H), 1.69-1.60 (m, 1 H). Step K. Synthesis of (2R,3S,5R,6S)-5-azido-2-(azidomethyl)-3-benzyloxy-6-(p- tolylsulfanyl)tetrahydropyran [00335] DIAD (3.45

of [(2R,3S,5R,6S)-5-azido-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methanol (4.5 g, 11.7 mmol) and triphenylphosphine (4.59 g, 17.5 mmol) in THF (90 mL) at 0 °C. The mixture was stirred at 0 °C for 20 min, then Et₃N (3.25 mL, 23.3 mmol) and DPPA (3.77 mL, 17.5 mmol) were added, and stirred at rt for 12 h. Silica was added, and the mixture was concentrated under reduced pressure. The material was purified on a silica gel column (dry loading, 120 g cartridge) with hexane and EtOAc (0-50%) to provide the title compound (3.61 g, 75%) as an oil. m/z (ESI) no ionization, HPLC (A05) tR= 2.94 m. Step L. Synthesis of (3R,5S,6R)-3-azido-6-(azidomethyl)-5-benzyloxy-tetrahydropyran-2-ol (Ring A1) [00336] Solid NBS (2.5

-5-azido-2-(azidomethyl)-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran (4 g, 9.74

at 0 °C. The mixture was stirred at 0 °C for 30 min, then diluted by 1:1 sat. NaHCO₃/Na₂S₂O₃ (60.0 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed brine (20 mL), dried (MgSO₄), filtered and concentrated under reduced pressure. The material was purified on silica gel (100 g high-capacity column) with hexanes AttyDktNo. R1119431100WO and EtOAc (0-60%) to provide the title compound (2.79 g, 94%) as a diastereomeric mixture. m/z (ESI); no ionization, HPLC A(205 nm) tR=2.36 m and 2.41 m. Example 2. Synthesis of (1R,2R,3S,4R,5S,6S)-3,5-bis(((benzyloxy)carbonyl)amino)-6- hydroxycyclohexane-1,2,4-triyl triacetate (Ring B1). (Ring B1) Step A: Synthesis of (((2R,3R,4R,5S)-3-(((2R,3S,4S,5R,6S)-4,5-

dihydroxy-6-(hydroxymethyl)-3- 2H-pyran-2-yl)oxy)-4-hydroxy-4- (hydroxymethyl)-5-methyltetrahydrofuran-2-yl)oxy)cyclohexane-1,2,4-triol

[00337] Dihydrostreptomycin sulfate (50 g), Ba(OH)₂-8H₂O (4 equivalents) and water (500 mL) were combined and heated under reflux for 2 days. After cooling to room temperature, CO₂ was bubbled through the reaction until it became slightly acidic (pH 6-7). The resulting mixture was filtered through diatomaceous earth, washing twice with water (75 mL). The aqueous layer was concentrated in vacuo and the aqueous solution used directly without further purification. 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 AttyDktNo. R1119431100WO

mL) over ~1h at room temperature. The reaction was stirred at room temperature for 2 days. Saturated aqueous sodium bicarbonate was added (50 mL) and the product extracted into EtOAc (2x75 mL) and washed with brine (50 mL). The combined organics were dried over sodium sulfate and the solvent removed in vacuo. The crude product was dissolved in 2-butanone (80 ml) and dripped slowly into MTBE (750 mL). The precipitated solids were collected by filtration and dried in a vacuum oven at 50°C to afford the titled product (52.1 g) as an off- white solid (72% purity by absorbance at 210 nm). 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

[00339] The crude compound from Step B (52.1 g) was dissolved in EtOAc (210 mL) and DMAP (721 mg, 0.1 eq) added at room temperature. DIPEA (103 mL) and Ac₂O (51 mL) in EtOAc (80 mL) was added to the reaction mixture dropwise over 1 h at room temperature. The reaction was stirred at room temperature overnight. The reaction was transferred to a separatory funnel with EtOAc (50mL, 0.9V) and washed with water (50 mL), sodium bicarbonate solution (2x50 mL), and brine (50 mL). The organics were dried over sodium sulfate and filtered to afford the titled products as a cream-colored foam (70 g; 63% AP at 210 nm). Step D: Synthesis of (1S,2S,3R,4S,5R,6R)-3,5-bis(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexane- 1,2,4-triyl triacetate (Ring B1) AttyDktNo. R1119431100WO

, (29.4 mL, 2.1 eq), ^{and BF} ₃ ^.Et ₂ ^{O (16.3 mL, 2.2 eq). The reaction was heated under reflux for 2 hours. 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 afford the titled product. The crude product was triturated with heptane (300 mL) to afford the titled product (23.9 g, 71% yield). Example 3. Synthesis of 2R,3R)-4-amino-N-[(1R,2S,3R,4R,5S,6S)-5-amino-3-[(2S,3R,4S,5R)-4- [(2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxyoxan-2-yl]oxy-3-hydroxy-5- (hydroxymethyl)oxolan-2-yl]oxy-4-[(2R,3R,5S,6R)-3-amino-6-(aminomethyl)-5-hydroxyoxan-2-yl]oxy- 2,6-dihydroxycyclohexyl]-3-fluoro-2-hydroxybutanamide (Compound 1) (Compound 1)

Step A. Synthesis of [(3R,5S,6R)-3-azido-6-(azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl] (1E)-2,2,2-trifluoro-N-phenyl-ethanimidate

AttyDktNo. R1119431100WO [00341] Cs₂CO₃ (5.55 g, 17 mmol) was added to a mixture of (3R,5S,6R)-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-ol (Ring A1; Example 1, Step L; 2.59 g, 8.51 mmol) and 2,2,2-Trifluoro-N- phenylacetimidoyl chloride (2.31 mL, 14.5 mmol) in dry DCM (50 mL). The mixture was stirred at rt for 2 h, then filtered (Diatomaceous earth) and concentrated under reduced pressure. The residue was purified on silica gel (dry loading, 200 g high-capacity column) using hexane and EtOAc (0-40%) to provide title compound (3.63 g, 90%) as an oil. m/z (ESI): no ionization, HPLC (A05) tR=2.94 m. Step B. Synthesis of [(1S,2R,3S,4S,5R,6S)-3,4-diacetoxy-5-[(3R,5S,6R)-3-azido-6- (azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-2,6- bis(benzyloxycarbonylamino)cyclohexyl] acetate

[00342] A [ - - 2-yl] (1E)-2,2,2- trifluoro-N-phenyl-ethanimidate (3.63 g, 0.505 mmol) and [(1S,2R,3S,4S,5R,6R)-3,4-diacetoxy-2,6- bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (Ring B1, Example 2, Step C; 6.56 g, 11.5 mmol) were co-evaporated with anhydrous toluene (2 x 12 mL). The mixture was dried under reduced pressure for 30 min. The residue was taken in anhydrous DCM (60 mL) and Et₂O (30 mL), then transferred via a syringe to a RBF charged with pre-activated molecular sieves (3Å and 4Å). The mixture was stirred at rt for 10 min, then Bi(OTf)₃ (5.51 g, 8.4 mmol) was added in one portion at 0 °C and the mixture was stirred at 0 °C for 0.5 h, then at rt for 18h. The mixture was diluted by saturated aq. NH₄Cl solution (50 mL), and DCM (200 mL), then filtered (diatomaceous earth) and partitioned. The aqueous layer was extracted with DCM (3 x 50 mL). The combined organic layers were washed brine (10 mL), dried (MgSO₄), filtered and concentrated under reduced pressure. The material was purified by C18 column using 10-100% MeCN in 10 mM ammonium formate (pH 3.8) and lyophilized to give the title product (5.9 g, 84%, α/β = 3:1) as an anomeric mixture. m/z (ESI) [M+NH₄]⁺: 876.3, HPLC (A05) tR= 2.78 m. Step C. Synthesis of Benzyl N-[(1S,2R,3R,4S,5R,6R)-2-[(3R,5S,6R)-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-yl]oxy-5-(benzyloxycarbonylamino)-3,4,6-trihydroxy- cyclohexyl]carbamate AttyDktNo. R1119431100WO [00343]

- 3,4-diacetoxy-5-[(3R,5S,6R)-3-azido-6-(azidomethyl)-5-benzyloxy-tetra hydropyran-2-yl]oxy-2,6- bis(benzyloxycarbonylamino)cyclohexyl] acetate (5.7 g, 6.64 mmol) in MeOH (300 mL) at rt. The mixture was stirred at rt for 4.5 h, diluted by AcOH (0.98 mL) until pH ~5, then concentrated under reduced pressure. The solid was diluted with EtOAc (150 mL) and water (30 mL), then partitioned. The aqueous was extracted with EtOAc (3 x 50 mL). The combined organic phase was washed by brine (2 x 50 mL), dried (MgSO₄), filtered, and concentrated under reduced pressure to give the title product (4.7 g, 97%) as a solid (anomeric mixture). This solid was used directly to the next step without further purification. m/z (ESI) [M+H]⁺ 733.2, HPLC A(05) tR=2.53 m. Step E. Synthesis of Benzyl N-[(3aS,4R,5S,6S,7R,7aS)-4-[(2R,3R,5S,6R)-3-azido-6- (azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-7-(benzyloxycarbonylamino)-6-hydroxy- spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1'-cyclohexane]-5-yl]carbamate

[00344] A mixture of benzyl N-[(1S,2R,3R,4S,5R,6R)-2-[(3R,5S,6R)-3-azido-6-(azidomethyl)-5-benzyloxy- tetrahydropyran-2-yl]oxy-5-(benzyloxycarbonylamino)-3,4,6-trihydroxy-cyclohexyl] carbamate (4.7 g, 6.41 mmol, anomeric mixture), 1,1-dimethoxycyclohexane (5.9 mL, 38.5 mmol) and PPTS (484 mg, 1.92 mmol, dried before using) in dry DME (150 mL) was stirred at 90 °C for 6 h (about 5% starting material remained). The mixture was cooled and concentrated under reduced pressure. The material was purified by reversed phase chromatography (ACCQ, BEH C1830x100 mm) using a gradient of 65%-75% ammonium formate (10 mM, pH 3.8) in MeCN to provide the title compound (3.25 g, 62%) as a solid. m/z (ESI) [M+H]⁺: 813.3, HPLC (A05) tR= 2.89 m. AttyDktNo. R1119431100WO Step F. Synthesis of Benzyl N-[(3aS,4R,5S,6S,7R,7aS)-4-[(2R,3R,5S,6R)-3-azido-6- (azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-6-benzyloxy-7- (benzyloxycarbonylamino)spiro-[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1'-cyclohexane]- 5-yl]carbamate [00345] NaH

of benzyl N- [(3aS,4R,5S,6S,7R,7aS)-4-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)-5-benzyloxy-tetrahydropyr-an-2- yl]oxy-7-(benzyloxycarbonylamino)-6-hydroxy-spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1'- cyclohexane]-5-yl]carbamate (2.8 g, 3.44 mmol) and BnBr (4.6 mL, 38.6 mmol) in THF (100 mL) at 0 °C. The mixture was stirred at 50 °C for 18 h. The mixture was cooled to rt, then AcOH (0.79 mL, 13.8 mmol) was added followed by EtOAc (100 mL) and silica gel. The mixture was concentrated under reduced pressure. The solid was purified on silica gel column (120 g, dry loading) using hexanes and EtOAc (0-100%) to provide the title product (970 mg, 31%) as a solid. m/z (ESI) [M+H]⁺ 903.3; HPLC (A05); tR = 3.02 m. Step G. Synthesis of Benzyl N-[(1S,2R,3R,4S,5S,6R)-2-[(2R,3R,5S,6R)-3-azido-6- (azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-6-benzyloxy-5- (benzyloxycarbonylamino)-3,4-dihydroxy-cyclohexyl]carbamate

[00346] A solution of benzyl N-[(3aS,4R,5S,6S,7R,7aS)-4-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-yl]oxy-6-benzyloxy-7-(benzyloxycarbonylamino)spiro[3a,4,5,6,7,7a- hexahydro-1,3-benzodioxole-2,1'-cyclohexane]-5-yl]carbamate (970 mg, 1.07 mmol) in THF (14.5 mL), water (4.84 mL) and AcOH (12.1 mL) was heated at 80 °C for 8 h, then cooled to rt. Water (100 mL) was added, and the suspension was filtered on a medium glass frit. The solid was rinsed with water (100 mL) and dried under vacuum to provide the title product (830 mg, 94%) as an off-white solid. m/z (ESI) [M+H]⁺: 823.4, HPLC (A05) tR = 2.75 m. AttyDktNo. R1119431100WO Step H. Synthesis of Benzyl N-[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-yl]oxy-4-benzyloxy-7-hydroxy-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3- benzoxazol-5-yl]carbamate

[00347] mg, was a of benzyl N- [(1S,2R,3R,4S,5S,6R)-2-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-6- benzyloxy-5-(benzyloxycarbonylamino)-3,4-dihydroxy-cyclohexyl]carbamate (830 mg, 1.01 mmol) in dry DMF (10 mL) at 0 °C. The mixture was stirred at rt for 2 h, then AcOH (0.89 mL, 15.5 mmol) was added followed by water (100 mL). The suspension was filtered, the solid was rinsed with water (100 mL) and lyophilized to provide the title compound (720 mg, quant.) as a solid. m/z (ESI) [M+H]⁺: 715.2, HPLC (A05) tR = 2.67 m. Step I. Synthesis of N-[(1S,2R,3R,4S,5S,6R)-5-amino-2-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-yl]oxy-3-[(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)tet -rahydrofuran-2-yl]oxy-6- benzyloxy-4-hydroxy-cyclohexyl]carbamate

azido-6- (azidomethyl)tetrahydropyran-2-yl]oxy-5-hydroxy-tetrahydrofuran-2-yl]methyl acetate; WO2019194858; 1.9 g, 3.58 mmol) and K₂CO₃ (812 mg, 5.88 mmol) in dry DCM (50 mL) was added CCl₃CN (0.56 mL, 5.88 mmol). The mixture was stirred at rt 24 h, then filtered on diatomaceous earth, rinsed with DCM and concentrated under reduced pressure. Benzyl N-[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,5S,6R)-3-azido-6- (azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-4-benzyloxy-7-hydroxy-2-oxo-3a,4,5,6,7,7a- hexahydro-3H-1,3-benzoxazol-5-yl]carbamate (700 mg, 0.979 mmol) in dry DCM (48 mL) was added to the AttyDktNo. R1119431100WO previous crude material, then transferred to a RBF charged with pre-activated 3Å and 4Å sieves. The mixture was cooled to -78 °C, then BF_{3 ^}OEt₂ (2 mL, 16.2 mmol) was added slowly at -78 °C. Upon completion of the addition, the dry ice bath was removed immediately. 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 (100 mL). The separated aqueous layer was extracted with DCM (3 x 20 mL). The combined organic layer was washed with brine (15 mL), dried (MgSO₄), filtered and concentrated under reduced pressure. The residue was taken in 1,4-dioxane (12 mL) and water (6.0 mL), then LiOH ^H₂O (1233 mg, 29.4 mmol) was added, and the mixture was stirred at 40 °C for 18 h. The mixture was cooled to rt, diluted by water (10 mL) and EtOAc (100 mL), and then partitioned. The separated aqueous layer was extracted with EtOAc (4 x 20 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 48-65% MeCN in 10 mM ammonium bicarbonate (pH 10) to provide the title product (500 mg, 49% over 2 steps) as a solid. m/z (ESI) [M+H]⁺: 1033.5, HPLC (A05) tR= 2.38 m. Step J. Synthesis of Benzyl N-[(1S,2R,3R,4S,5S,6R)-2-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)- 5- benzyloxy-tetrahydropyran-2-yl]oxy-3-[(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-[[(2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy] - butanoyl]amino]-6-benzyloxy-4-hydroxy-cyclohexyl]carbamate

- - azido-6- (azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-3-[(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)tet -rahydrofuran-2- yl]oxy-6-benzyloxy-4-hydroxy-cyclohexyl]carbamate (250 mg, 0.242 mmol) and (2R,3R)-4-azido-3-fluoro- 2-[(4-methoxyphenyl)methoxy]butanoic acid (118 mg, 0.415 mmol) in anhydrous THF (12 mL) was added DMTMM chloride (118 mg, 0.425 mmol). The mixture was stirred at rt for 18 h, then diluted by a saturated aqueous NH₄Cl solution (5 mL) and EtOAc (30 mL). The separated aqueous layer was extracted with EtOAc (3 x 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 prep HPLC (ACCQ, BEH C1830x100) AttyDktNo. R1119431100WO using a gradient of 58-75% MeCN in Ammonium formate (10 mM, pH 4) and lyophilized to provide title product (252 mg, 80%) as a white solid. m/z (ESI) [M+H]⁺: 1298.4, HPLC A(05) tR=2.82 m. Step K. Synthesis of 2R,3R)-4-amino-N-[(1R,2S,3R,4R,5S,6S)-5-amino-3-[(2S,3R,4S,5R)-4- [(2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxyoxan-2-yl]oxy-3-hydroxy-5- (hydroxymethyl)oxolan-2-yl]oxy-4-[(2R,3R,5S,6R)-3-amino-6-(aminomethyl)-5-hydroxyoxan- 2-yl]oxy-2,6-dihydroxycyclohexyl]-3-fluoro-2-hydroxybutanamide (Compound 1)

[00350] A solution of benzyl N-[(1S,2R,3R,4S,5S,6R)-2-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-yl]oxy-3-[(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)tetrahydro-furan-2-yl]oxy-5-[[(2R,3R)-4- azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoyl]amino]-6-benzyloxy-4-hydroxy- cyclohexyl]carbamate (252 mg, 0.194 mmol) in MeOH (6 mL) was added to a suspension of Pd(OH)₂ (980 mg, 1.40 mmol, 20% wet) in a mixture of AcOH (5 mL) and water (2 mL), degassed by 3 x vacuum-H₂ cycle, then bubbled with hydrogen for 10 min. The mixture was stirred under hydrogen (1 atm, refilling the balloon with H₂ when necessary) for 4 h, then degassed with N₂ for 15 min. The mixture was filtered on 0.22 μM PVDF filter syringe which was washed with MeOH (3 x 10 mL) and water (3 x 10 mL), and the volatiles were evaporated under reduced pressure. The material was purified by prep-HPLC using 5% MeCN in 10 mM NH₄HCO₃ (pH 10) and lyophilized to give a solid. H₂SO₄ (0.05 M in H₂O) was added dropwise to a solution of the above solid in H₂O (3 mL) at 0 °C until pH 4-5, then lyophilized to give a solid, which was dissolved in water (2 mL), followed by MeOH (18 mL) to give a precipitate. The solid was filtered through 0.22 uM PVDF syringe filter and rinsed with MeOH (2 x 10 mL). The solid was dissolved with water (25 mL), then concentrated under reduced pressure. The resulting solid was dissolved in water (2 mL) and lyophilized to give Compound 1 (113 mg, 57%) as a white solid.¹H NMR (500 MHz, D₂O) δ 5.82 (d, J = 3.9 Hz, 1H), 5.33 (d, J = 1.9 Hz, 1H), 5.29 – 5.15 (m, 2H), 4.53 – 4.38 (m, 2H), 4.35 (dd, J = 4.7, 1.9 Hz, 1H), 4.26 (ddd, J = 6.9, 3.9, 1.6 Hz, 1H), 4.21 – 4.11 (m, 2H), 4.10 – 4.00 (m, 1H), 3.95 – 3.83 (m, 4H), 3.79 – 3.75 (m, 1H), 3.73 – 3.56 (m, 5H), 3.55 – 3.49 (m, 1H), 3.49 – 3.27 (m, 5H), 3.21 (t, J = 10.2 Hz, 1H), 3.13 (dd, J = 13.6, 8.0 Hz, AttyDktNo. R1119431100WO 1H), 2.22 (dt, J = 11.7, 4.4 Hz, 1H), 1.98 (q, J = 12.2 Hz, 1H), exchangeable protons are not observed. m/z (ESI) [M+H]⁺: 734.3, HPLC (A05) tR= 0.26 m. Example 4. Synthesis of (2S)-4-amino-N-[(1R,2S,3R,4R,5S,6S)-5-amino-4-[(2R,3R,5S,6R)-3- amino-6-(aminomethyl)-5-hydroxyoxan-2-yl]oxy-3-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3- amino-6-(aminomethyl)-4,5-dihydroxyoxan-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)oxolan-2- yl]oxy-2,6-dihydroxycyclohexyl]-2-hydroxybutanamide (Compound 2) OH OH H H₂N N NH_{2 2} (Compound 2) Step A. Synthesis of

-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-yl]oxy-3-[(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-6-benzyloxy-5-[[(2S)-2-benzyloxy-4- (benzyloxycarbonylamino)butanoyl]amino]-4-hydroxy-cyclohexyl]carbamate [00351] To a solution of benzyl N-[(1S,2R,3R,4S,5S,6R)-5-amino-2-[(2R,3R,5S,6R)-3-azido-6- (azidomethyl)-5-benzyloxy-tetrahydropyran-2-yl]oxy-3-[(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-6-benzyloxy-4-hydroxy-cyclohexyl]carbamate (Example 3, Step I; 250 mg, 0.242 mmol), (2S)-2- benzyloxy-4-(benzyloxycarbonylamino)butanoic acid (139 mg, 0.405 mmol) and DIPEA (0.14 mL, 0.802 mmol) in anhydrous DMF (10 mL) was added PyBOP (168 mg, 0.323 mol) at 0 °C. The mixture was stirred at 0 °C for 1 h, and at rt for 18 h, then poured into water (5 mL). The pH was adjusted to 4 using 1N HCl and then EtOAc (40 mL) was added. The separated aqueous layer was extracted with EtOAc (4x 15 mL). The combined organic layers were washed with brine (5 mL), dried (MgSO₄), filtered and concentrated under reduced pressure. The material was purified by reversed phase column (ACCQ, BEH C1830x100) using a gradient of 58-75% MeCN in ammonium formate (10 mM, pH 4) and lyophilized to provide the title compound (256 mg, 78%) as a white solid. m/z (ESI) [M+H]⁺:1359.3, HPLC A(05), tR=2.86 m. AttyDktNo. R1119431100WO Step B. Synthesis of (2S)-4-amino-N-[(1R,2S,3R,4R,5S,6S)-5-amino-4-[(2R,3R,5S,6R)-3-amino- 6-(aminomethyl)-5-hydroxyoxan-2-yl]oxy-3-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-amino-6- (aminomethyl)-4,5-dihydroxyoxan-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)oxolan-2-yl]oxy-2,6- dihydroxycyclohexyl]-2-hydroxybutanamide (Compound 2) [00352] A solution of benzyl N-[(1S,2R,3R,4S,5S,6R)-2-[(2R,3R,5S,6R)-3-azido-6-(azidomethyl)-5- benzyloxy-tetrahydropyran-2-yl]oxy-3-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidom -ethyl)-4,5- dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-6-benzyloxy-5- [[(2S)-2-benzyloxy-4-(benzyloxycarbonylamino)butanoyl]amino]-4-hydroxy-cyclohexyl]carbamate (256 mg, 0.188 mmol) in MeOH (6 mL) was added to a suspension of Pd(OH)₂ (980 mg, 1.4 mmol, 20% wet) in a mixture of AcOH (5 mL) and water (2 mL), degassed by 3 x vacuum-H₂ cycle, then bubbled with hydrogen for 10 min. Hydrogen was bubbled through the reaction mixture for 3.5 h. After completion, N2 was bubbled through the reaction mixture for 15 min, then the mixture was filtered on 0.22 μM PVDF filter syringe which was washed with MeOH (3 x 10 mL) and water (3 x 10 mL), and the volatiles were evaporated under reduced pressure. The material was purified by filtration using 5% MeCN in 10 mM NH₄HCO₃ (pH 10) and lyophilized to give a solid. Sulfuric acid (0.05M in H₂O) was added dropwise to a solution of the above solid in H₂O (3 mL) at 0 °C until pH 4-5, then lyophilized to give a solid, which was dissolved in water (2 mL), followed by MeOH (18 mL) to give a precipitate. This solid was filtered through 0.22 uM PVDF syringe filter, rinsed with MeOH (2 x 10 mL), washed with water (25 mL), then concentrated under reduced pressure. The resulting solid was dissolved in water (2 mL) and lyophilized to give Compound 2 (90.8 mg, 48%) as a white solid.¹H NMR (500 MHz, D₂O) δ 5.83 (d, J = 3.6 Hz, 1H), 5.33 (d, J = 1.9 Hz, 1H), 5.23 (d, J = 2.0 Hz, 1H), 4.48 (dd, J = 7.3, 4.7 Hz, 1H), 4.37 (dd, J = 4.7, 2.1 Hz, 1H), 4.32 (dd, J = 8.0, 4.1 Hz, 1H), 4.29 – 4.25 (m, 1H), 4.18 (t, J = 3.2 Hz, 1H), 4.15 (ddd, J = 7.7, 5.2, 3.0 Hz, 1H), 4.08 (t, J = 10.0 Hz, 1H), 3.93 – 3.82 (m, 4H), 3.76 (d, J = 3.5 Hz, 1H), 3.74 – 3.56 (m, 5H), 3.54 – 3.51 (m, 1H), 3.42 – 3.34 (m, 2H), 3.33 – 3.20 (m, 2H), 3.16 – 3.05 (m, 3H), 2.28 – 2.18 (m, 1H), 2.18 – 2.07 (m, 1H), 2.05 – 1.92 (m, 2H), exchangeable protons are not observed. m/z (ESI) [M+H]⁺: 716.3, HPLC (A05) tR= 0.24 m. Example 5. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3- amino-6-((S)-1-aminoethyl)-5- 2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4- (((2R,3R,4R,5S,6S)-3-amino-6-

-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) AttyDktNo. R1119431100WO OH OH H H₂N N NH_{2 2} (Compound 3) Step A.

-6-(p-tolylthio)tetrahydro-2H-pyran-2- carbaldehyde [00353] Dess-Martin periodinane (36.5 g, 81.7 mmol) was added to a mixture of [(2R,3S)-5-azido-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methanol (30.0 g, 77.8 mmol) and NaHCO₃ (19.6 g, 233 mmol) in dry DCM (389 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h, and at 22 °C for 2 h. The mixture was diluted with sat. aq. Na₂S₂O₃ (400 mL) and sat. aq. NaHCO₃ (400 mL) at 0 °C. The mixture was stirred at 0 °C for 5 min, and at 22 °C for 15 min. The aqueous phase was extracted with DCM (3 x 200 mL), and the combined organic extracts were washed with sat. aq. Na₂S₂O₃ (300 mL), sat. aq. NaHCO₃ (300 mL), brine (300 mL), dried (Na₂SO₄), filtered, and concentrated to provide the title compound as an oil (29.7 g, 98%). ¹H NMR (300 MHz, DMSO-d₆) δ 9.61 (d, J = 1.4 Hz, 1H), 7.40 (d, J = 8.0 Hz, 2H), 7.35 – 7.26 (m, 5H), 7.19 (d, J = 8.0 Hz, 2H), 4.91 (d, J = 9.8 Hz, 1H), 4.61 (d, J = 11.7 Hz, 1H), 4.45 (d, J = 11.6 Hz, 1H), 4.20 – 4.01 (m, 1H), 3.69 (td, J = 10.1, 4.4 Hz, 1H), 3.52 (td, J = 10.8, 4.6 Hz, 1H), 2.71 (dd, J = 11.9, 4.5 Hz, 1H), 2.29 (s, 3H), 1.66 (q, J = 11.3 Hz, 1H); m/z (ES+) [M+H₂O]⁺ = 401.1; HPLC (A05) t_R = 1.67 min. Step B. Synthesis (S)-N-((E)-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H-pyran-2- yl)methylene)-2-methylpropane-2-sulfinamide [00354] Anhydrous CuSO₄ (28.8 g, 181 mmol) was added to a mixture of (2S,3S,5R)-5-azido-3-benzyloxy-6- (p-tolylsulfanyl)tetrahydropyran-2-carbaldehyde (29.7 g, 76.0 mmol) and (S)-2-methylpropane-2-sulfinamide (27.6 g, 228 mmol) in dry DCM (290 mL) at 22 °C. The mixture was stirred at 22 °C for 44 h. The mixture was diluted with DCM (300 mL), filtered through Celite, washed with DCM (2 x 100 mL), and the filtrate was concentrated. The residue was diluted with water (500 mL), sat. aq. NH₄Cl (250 mL) and EtOAc (300 mL). The aqueous phase was extracted with EtOAc (3 x 200 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (200 mL), sat. aq. NaHCO₃ (200 mL), brine (200 mL), dried (Na₂SO₄), filtered and concentrated. The residue was purified by silica gel chromatography (220 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-20%) to provide the title compound as an oil (23.4 g, 59%). ¹H NMR (300 MHz, DMSO-d₆) δ 7.90 (d, J = 4.3 Hz, 1H), 7.43 – 7.37 (m, 2H), 7.36 – 7.23 (m, 5H), 7.18 – 7.10 (m, 2H), 4.90 (d, J = 9.9 Hz, 1H), 4.61 (d, J = 11.8 Hz, 1H), 4.47 – 4.33 (m, 2H), 3.64 – 3.46 (m, 2H), 2.75 (dt, J = 11.7, 4.5 Hz, 1H), 2.29 (s, 3H), 1.64 (q, J = 11.4 Hz, 1H), 1.12 (s, 9H). Note: Contains 7 wt% EtOAc; m/z (ES+) [M+H]⁺ = 487.1; HPLC (A05) t_R = 1.90 min. AttyDktNo. R1119431100WO Step C. Synthesis of (S)-N-((1S)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H- pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide [00355] MeMgBr (20.0 mL, 60.0 mmol, 3.00 mol/L in Et₂O) was added to a mixture of (NE,S)-N- [[(2R,3S,5R,6S)-5-azido-3-benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methylene]-2-methyl- propane-2-sulfinamide (9.03 g, 18.6 mmol) in dry DCM (127 mL) at -78 °C. The mixture was stirred at - 78 °C for 3 h, at -55 °C for 2 h. The mixture was diluted with sat. aq. NH₄Cl (25.0 mL) and EtOAc (125 mL) at -55 °C and was stirred for 30 min. The mixture was diluted with water (80.0 mL), sat aq. NH₄Cl (80.0 mL), EtOAc (125 mL). The aqueous phase was extracted with EtOAc (3 x 90.0 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (175 mL), brine (175 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (330 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-30%) to provide the title compound as a solid (5.35 g, 57%). ¹H NMR (300 MHz, DMSO-d₆) δ 7.44 – 7.40 (m, 2H), 7.39 – 7.23 (m, 5H), 7.18 (d, J = 8.0 Hz, 2H), 4.72 (d, J = 10.0 Hz, 1H), 4.56 (q, J = 10.8 Hz, 2H), 4.42 (d, J = 8.6 Hz, 1H), 3.76 – 3.58 (m, 2H), 3.47 – 3.34 (m, 2H), 2.80 – 2.71 (m, 1H), 2.30 (s, 3H), 1.48 (q, J = 11.5 Hz, 1H), 1.17 (d, J = 6.9 Hz, 3H), 1.07 (s, 9H); m/z (ES+) [M+H]⁺ = 503.2; HPLC (A05) t_R = 5.88 min. Step D. Synthesis of (S)-N-((1S)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H- pyran-2-yl)ethyl)-N-benzyl-2-methylpropane-2-sulfinamide [00356] KOtBu (4.24 g, 37.8 mmol) was added to a mixture of (R)-N-[(1S)-1-[(2R,3S,5R,6S)-5-azido-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (6.12 g, 12.2 mmol) and BnBr (4.76 mL, 40.1 mmol) in dry DMF (51.9 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h, and at 22 °C for 2 h. The mixture was diluted with AcOH (2.29 mL, 40.0 mmol), EtOAc (100 mL), water (100 mL), and sat. aq. NH₄Cl (200 mL) at 22 °C. The aqueous phase was extracted with EtOAc (3 x 80.0 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (175 mL), brine (175 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (100 g high- capacity cartridge) using a gradient of EtOAc in hexanes (0-25%) to provide the title compound as a solid (5.69 g, 79%). ¹H NMR (400 MHz, DMSO-d₆) δ 7.43 – 7.39 (m, 4H), 7.35 – 7.26 (m, 3H), 7.25 – 7.18 (m, 5H), 7.17 – 7.14 (m, 2H), 4.64 (d, J = 10.0 Hz, 1H), 4.56 (d, J = 11.2 Hz, 1H), 4.28 (d, J = 11.2 Hz, 1H), 4.16 – 4.04 (m, 2H), 3.56 (td, J = 9.9, 4.6 Hz, 1H), 3.50 – 3.41 (m, 1H), 3.36 (dd, J = 7.2, 4.1 Hz, 1H), 3.30 – 3.26 (m, 1H), 2.82 (dt, J = 11.8, 4.6 Hz, 1H), 2.29 (s, 3H), 1.46 (q, J = 11.5 Hz, 1H), 1.18 (d, J = 7.1 Hz, 3H), 1.14 (s, 9H); m/z (ES+) [M+H]⁺ = 593.4; HPLC (A05) t_R = 2.01 min. Step E. Synthesis of benzyl ((1S)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H- pyran-2-yl)ethyl)(benzyl)carbamate [00357] HCl (24.0 mL, 24.0 mmol, 1.0 N in H₂O) was added to a mixture of (S)-N-[(1S)-1-[(2R,3S,5R,6S)-5- azido-3-benzyloxy-2-methyl-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]ethyl]-N-benzyl-2-methyl-propane-2- sulfinamide (11.9 g, 20.1 mmol) in dioxane (105 mL) at 22 °C. The mixture was stirred at 22 °C for 90 min. m/z (ES+) [M+H]⁺ = 489.1; HPLC (A05) t_R = 2.00 min. The mixture was diluted with PBS buffer (254 mL) AttyDktNo. R1119431100WO to pH ≈ 7.6, and CbzCl (5.70 mL, 40.1 mmol) was added to the mixture at 22 °C. The mixture was stirred at 22 °C for 3 h and was diluted with EtOAc (150 mL). The aqueous phase was extracted with EtOAc (3 x 100 mL), and the combined organic extracts were washed with brine (300 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (200 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-15%) to provide the title compound as an oil (12.1 g, 97%). m/z (ES+) [M+H]⁺ = 623.4; HPLC (A05) t_R = 2.12 min. Step F. Synthesis of benzyl ((1S)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-hydroxytetrahydro-2H-pyran- 2-yl)ethyl)(benzyl)carbamate [00358] NBS (3.97 g, 22.3 mmol) was added to a mixture of benzyl N-[(1S)-1-[(2R,3S,5R,6S)-5-azido-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (12.1 g, 19.4 mmol) in acetone (400 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h, was diluted with sat. aq. NaHCO₃ (300 mL), sat. aq. Na₂S₂O₃ (300 mL) and water (100 mL). The aqueous phase was extracted with EtOAc (4 x 100 mL), and the combined organic extracts were washed with brine (300 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (330 g high-capacity cartridge) using a gradient of EtOAc and hexanes (0-30%) to provide the title compound as an oil (8.09 g, 81%). m/z (ES+) [M+H]⁺ = 517.4 and m/z (ES+) [M+H-H₂O]⁺ = 499.4; HPLC (A05) t_R = 1.84 min. Step G. Synthesis of (3R,5S,6R)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)-5- (benzyloxy)tetrahydro-2H-pyran-2-yl (E)-2,2,2-trifluoro-N-phenylacetimidate [00359] Cs₂CO₃ (9.69 g, 29.8 mmol) was added to a mixture of benzyl N-[(1S)-1-[(2R,3S,5R)-5-azido-3- benzyloxy-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (8.09 g, 15.7 mmol), 2,2,2-trifluoro- N-phenyl-acetimidoyl chloride (4.40 mL, 27.6 mmol) in dry DCM (86.7 mL) at 22 °C. The mixture was stirred at 22 °C for 18 h. The mixture was filtered through Celite, washing with DCM (3 x 80.0 mL), and the filtrate was concentrated. The residue was purified by silica gel chromatography (220 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-10%) to provide the title compound as an oil (9.75 g, 91%). m/z (ES+) [M+Na]⁺ = 710.4 and m/z (ES+) [M+H-H₂O]⁺ = 499.2; HPLC (A05) t_R = 2.13 min. Step H. Synthesis of dibenzyl ((1R,2R,3S,4R,5R,6S)-4-(((3R,5S,6R)-3-azido-6-((S)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-2,5,6- trihydroxycyclohexane-1,3-diyl)dicarbamate [00360] [(3R,5S,6R)-3-Azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]-5-benzyloxy- tetrahydropyran-2-yl] (1E)-2,2,2-trifluoro-N-phenyl-ethanimidate (9.75 g, 14.2 mmol), [(1S,2R,3S,4S,5R,6R)- 3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl] acetate (11.4 g, 19.8 mmol) were co- evaporated with dry toluene (2 x 50.0 mL) and dried under vacuum for 30 min charged with pre-activated 4Å molecular sieves (9.75 g) and dried under vacuum for 30 min. The residue was dissolved with dry 2-MeTHF (219 mL) at 0 °C. Bi(OTf)₃ (10.2 g, 15.6 mmol) was added to a mixture at 0 °C. The mixture was stirred at 0 °C for 2 h. The mixture was diluted with DCM (250 mL), filtered through Celite, washing with DCM (250 mL), and the filtrate was diluted with sat. aq. NH₄Cl (270 mL) and water (180 mL). The aqueous phase was extracted with DCM (3 x 150 mL), and the combined organic extracts were washed with brine (400 mL), dried AttyDktNo. R1119431100WO (Na₂SO₄), filtered, and concentrated. m/z (ES+) [M+H]⁺ = 1071.6; HPLC (A05) t_R = 1.99 min. The residue was dissolved in dry MeOH (219 mL) and NaOMe (2.45 mL, 11.3 mmol, 4.62 mol/L in MeOH) was added at 22 °C. The mixture was stirred at 22 °C for 2 h, was diluted with AcOH (2.43 mL, 42.5 mmol), and was concentrated. The residue was diluted with EtOAc (540 mL), filtered through Celite, washed with MeOH (540 mL), and the filtrate was concentrated. The residue was purified by silica gel chromatography (220 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-60%) to provide the title compound as a solid (4.68 g, 35% over 2 steps). m/z (ES+) [M+H]⁺ = 945.6; HPLC (A05) t_R = 1.85 min. Step I. Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((3R,5S,6R)-3-azido-6-((S)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-6- hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate [00361] PPTS (466 mg, 1.85 mmol) was added to a mixture of benzyl N-[(1S)-1-[(2R,3S,5R,6S)-5-azido-3- benzyloxy-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy- cyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (4.68 g, 4.95 mmol) and 1,1- dimethoxycyclohexane (4.00 mL, 26.3 mmol) in dry DME (53.5 mL) at 22 °C. The mixture was stirred at 90 °C for 4 h. The mixture was diluted with EtOAc (100 mL) and water (100 mL). The aqueous phase was extracted with EtOAc (3 x 50.0 mL), and the combined organic extracts were washed with sat. aq. NaHCO₃ (150 mL), sat. aq. NH₄Cl (150 mL), brine (150 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (100 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-40%) to provide the title compound as a solid (3.76 g, 74%). m/z (ES+) [M+H]⁺ = 1025.7; HPLC (A05) t_R = 2.08 min. Step J. Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((3R,5S,6R)-3-azido-6-((S)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-6- (benzyloxy)hexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate [00362] NaH (631 mg, 15.8 mmol, 60% dispersion in mineral oil) was added to a mixture of benzyl N-[(1S)- 1-[(2R,3S,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-3-benzyloxy- tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (3.76 g, 3.67 mmol) and BnBr (1.31 mL, 11.0 mmol) in dry THF (85.3 mL) at 0 °C. The mixture was stirred at 22 °C for 2 h, was diluted with AcOH (0.945 mL, 16.5 mmol), EtOAc (100 mL), and was concentrated. The residue was purified by silica gel chromatography (100 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-30%) to provide the title compound as a solid (3.35 g, 82%). m/z (ES+) [M+H]⁺ = 1115.7; HPLC (A05) t_R = 2.22 min. Step K. Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((3R,5S,6R)-3-azido-6-((S)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-2- (benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate [00363] AcOH (8.60 mL, 150 mmol) was added to a mixture of benzyl N-[(1S)-1-[(2R,3S,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-3-benzyloxy-tetrahydropyran-2-yl]ethyl]-N-benzyl- AttyDktNo. R1119431100WO carbamate (3.35 g, 3.00 mmol) in THF (13.5 mL) and water (5.60 mL) at 22 °C. The mixture was stirred at 80 °C for 24 h. AcOH (4.20 mL, 73.4 mmol) and water (2.80 mL) were added to the mixture and was stirred at 80 °C for 22 h and was diluted with EtOAc (150 mL) and sat. aq. NaHCO₃ (250 mL). The aqueous phase was extracted with EtOAc (3 x 100 mL), and the combined organic extracts were washed with sat. aq. NaHCO₃ (250 mL), brine (250 mL), dried (Na₂SO₄), filtered, and concentrated to provide the title compound as a solid (3.04 g, 98%). m/z (ES+) [M]⁺ = 1035.6; HPLC (A05) t_R = 1.97 min Step L. Synthesis of benzyl ((1S)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-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 [00364] NaOtBu (1.41 g, 14.7 mmol) was added to a mixture of benzyl N-[(1S)-1-[(2R,3S,5R,6S)-5-azido-3- benzyloxy-6-[(1R,2S,3R,4S,5S,6R)-3-benzyloxy-2,4-bis(benzyloxycarbonylamino)-5,6-dihydroxy- cyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (3.04 g, 2.94 mmol) and TEBA-Cl (3.34 g, 14.7 mmol) in dry DMF (49.8 mL) at 0 °C. The mixture was stirred at 0 °C for 30 min, diluted with sat. aq. NH₄Cl (300 mL), brine (150 mL) and EtOAc (300 mL). The aqueous phase was extracted with EtOAC (3 x 120 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (250 mL), sat. aq. NaHCO₃ (250 mL), brine (250 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (120 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-50%) to provide the title compound as a solid (2.28 g, 84%). m/z (ES+) [M+H]⁺ = 927.6; HPLC (A05) tR = 1.92 min. Step M. 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,5S,6R)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)- 5-(benzyloxy)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 [00365] K₂CO₃ (2.04 g, 14.8 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 (2S,3R,4R,5R,6R)-6-(((2R,3R,4R)-4-acetoxy-2-(acetoxymethyl)-5-hydroxytetrahydrofuran-3-yl)oxy)- 5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (3.37 g, 6.35 mmol) and CCl₃CN (1.48 mL, 14.8 mmol) in dry DCM (60.0 mL) at 22 °C. The mixture was stirred at 22 °C for 21 h, filtered on Celite, washing with DCM (3 x 30.0 mL), and the filtrate was concentrated. [00366] The residue (2.28 g, 2.46 mmol) was co-evaporated with dry toluene (3 x 25.0 mL) under reduced pressure. The mixture was dried under vacuum for 30 min. Crushed pre-activated 4Å molecular sieves (2.28 g) were added to the residue and was dissolved in dry DCM (60.0 mL) at -78 °C. BF₃ ^.OEt₂ (4.58 mL, 37.1 mmol) was added to the mixture at -78 °C and was stirred at 22 °C for 90 min. The mixture was diluted with sat. aq. NaHCO₃ (100 mL), filtered through Celite, washing with DCM (3 x 40.0 mL). The filtrate was diluted with sat. aq. NaHCO₃ (60.0 mL) and DCM (60.0 mL). The aqueous phase was extracted with DCM (3 x 80.0 mL), and the combined organic extracts were washed with sat. aq. NaHCO₃ (200 mL), brine (200 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (100 g high- AttyDktNo. R1119431100WO capacity cartridge) using a gradient of EtOAc in hexanes (0-50%) to provide the title compound as a solid (3.45 g, 97%). m/z (ES+) [M-CO]⁺ = 1411.9; HPLC (A05) t_R = 2.04 min. Step N. Synthesis of benzyl ((S)-1-((2R,3S,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-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy- 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3- hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate [00367] LiOH^.H₂O (3.02 g, 71.9 mmol) was added to a mixture of Rev-326 (3.45 g, 2.40 mmol) in dioxane (64.7 mL) and water (32.3 mL) at 22 °C. The mixture was stirred at 40 °C for 22 h and was diluted with EtOAc (100 mL), sat. aq. NH₄Cl (100 mL) and water (50.0 mL). The aqueous phase was extracted with EtOAc (3 x 80.0 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (100 mL), sat. aq. NaHCO3 (100 mL), brine (100 mL), dried (Na2SO4), filtered, and concentrated to provide the title compound as a solid (2.69 g, 90%). m/z (ES+) [M+H]⁺ = 1245.7; HPLC (A05) t_R = 1.79 min. Step O. Synthesis of benzyl ((S)-1-((2R,3S,5R,6R)-5-azido-6-(((1R,2R,3S,4S,5R,6S)-4-((2R,3R)-4-azido- 3-fluoro-2-((4-methoxybenzyl)oxy)butanamido)-2-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-azido-6- (azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3- hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate [00368] DMTMMCl (156 mg, 0.564 mmol) was added to a mixture of ((S)-1-((2R,3S,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- dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5- (benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro- 2H-pyran-2-yl)ethyl)(benzyl)carbamate (400 mg, 0.321 mmol) and (2R,3R)-4-azido-3-fluoro-2-[(4- methoxyphenyl)methoxy]butanoic acid (136 mg, 0.482 mmol) in dry 2-MeTHF (7.33 mL) at 22 °C. The reaction mixture was stirred at 22 °C for 21 h and was diluted with EtOAc (20.0 mL), sat. aq. NH₄Cl (35.0 mL), and water (35.0 mL). The aqueous phase was extracted with EtOAc (3 x 35.0 mL), and the combined organic extracts were washed with brine (50.0 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (12 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-90%) to provide the title compound as a solid (180 mg, 37%). m/z (ES+) [M-CD rings+H]⁺ = 1166.7; HPLC (A05) t_R = 1.98 min. Step P. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3-amino-6- ((S)-1-aminoethyl)-5-hydroxytetrahydro-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) [00369] Pd(OH)₂ (502 mg, 0.715 mmol, 20% purity) was added to a mixture of ((S)-1-((2R,3S,5R,6R)-5-azido- 6-(((1R,2R,3S,4S,5R,6S)-4-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanamido)-2- (((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2- AttyDktNo. R1119431100WO yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6- (((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro-2H-pyran-2- yl)ethyl)(benzyl)carbamate (180 mg, 0.119 mmol) in MeOH (4.25 mL), AcOH (3.64 mL) and water (1.52 mL) at 22 °C under nitrogen. The mixture was stirred at 22 °C under continuous bubbling of hydrogen balloon pressure for 5 h. The mixture was filtered through Celite, washing with water (15.0 mL), MeOH (30.0 mL) and water (15.0 mL). The filtrate was concentrated (rotavapor bath temperature < 35 °C) and was purified by preparative HPLC using 5% of MeCN in water ^10 mM Ammonium bicarbonate pH 10 ^, and was lyophilized. The solid was diluted with a solution of (NH₄)HCO₃ (1.02 g in 18.0 mL of water) at 22 °C. The mixture was stirred at 22 °C for 30 min, was filtered on frit (medium size), washed with distilled water (12.0 mL) and the filtrate was lyophilized. The solid was dissolved in water (15.0 mL), and (NH₄)₂SO₄ (94.5 mg, 0.715 mmol) was added at 22 °C. The mixture was stirred at 22 °C for 30 min and was filtered on a medium frit. The filtrate was added dropwise to MeOH (400 mL) and was stirred at 22 °C for 30 min. The suspension was centrifuged (rotation speed: 1765 rpm) for 30 min. MeOH supernatant was carefully removed, and the solid was dissolved in distilled water (20.0 mL). The solution was evaporated under reduced pressure to remove residual MeOH and lyophilized to provide the title compound (Compound 3) as a solid (36.0 mg, 29%). ¹H NMR (400 MHz, D₂O) δ 5.87 (d, J = 3.7 Hz, 1H), 5.44 – 5.36 (m, 1H), 5.36 – 5.18 (m, 2H), 4.62 – 4.43 (m, 2H), 4.40 (d, J = 4.9 Hz, 1H), 4.36 – 4.29 (m, 1H), 4.29 – 4.10 (m, 3H), 4.02 – 3.75 (m, 8H), 3.74 – 3.49 (m, 5H), 3.49 – 3.33 (m, 4H), 2.37 – 2.17 (m, 1H), 2.08 (p, J = 11.1 Hz, 1H), 1.42 (d, J = 6.7 Hz, 3H). Note: Missing hydrogens due to exchange with solvent. Example 6. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3- amino-6-((S)-1-aminoethyl)-5-hydroxytetrahydro-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 4) OH OH H N 2 (Compound 4) Step A. Synthesis of

-2-(((2S,3R,4S,5R)-4- (((2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy- 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-4-((S)-4- AttyDktNo. R1119431100WO (((benzyloxy)carbonyl)amino)-2-hydroxybutanamido)-3-hydroxycyclohexyl)oxy)-3- (benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate [00370] PyBOP (419 mg, 0.805 mmol) was added to a mixture of ((S)-1-((2R,3S,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- dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5- (benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro- 2H-pyran-2-yl)ethyl)(benzyl)carbamate (590 mg, 0.474 mmol), (2S)-4-(benzyloxycarbonylamino)-2- hydroxy-butanoic acid (180 mg, 0.711 mmol) and DIPEA (0.203 mL, 1.18 mmol) in dry DMF (9.00 mL) at 22 °C. The mixture was stirred at 22 °C for 30 min, and was diluted with EtOAc (30.0 mL), sat. aq. NH₄Cl (50.0 mL), and water (50.0 mL). The aqueous phase was extracted with EtOAc (3 x 50.0 mL), and the combined organic extracts were washed with brine (75.0 mL), dried (Na2SO4), filtered, and concentrated. The residue was purified by silica gel chromatography (40 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-100%) and a gradient of MeOH in DCM (0-5%) to provide the title compound as a solid (390 mg, 56%). m/z (ES+) [M-CD ring+H]⁺ = 1136.7; HPLC (A05) t_R = 1.88 min. Step B. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3-amino-6-((S)-1- aminoethyl)-5-hydroxytetrahydro-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 4) [00371] Pd(OH)₂ (555 mg, 0.790 mmol, 20% purity) was added to a mixture of ((S)-1-((2R,3S,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- dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5- (benzyloxy)-6-(((benzyloxy)carbonyl)amino)-4-((S)-4-(((benzyloxy)carbonyl)amino)-2- hydroxybutanamido)-3-hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro-2H-pyran-2- yl)ethyl)(benzyl)carbamate (390 mg, 0.263 mmol) in MeOH (9.40 mL), AcOH (8.00 mL) and water (3.40 mL) at 22 °C under nitrogen. The mixture was stirred at 22 °C under continuous bubbling of hydrogen ballon pressure for 4 h. The mixture was filtered through Celite, washed with water (50.0 mL), MeOH (50.0 mL) and water (50.0 mL). The filtrate was concentrated, was purified by preparative HPLC using 5% of MeCN in water ^10 mM Ammonium bicarbonate pH 10 ^, and was lyophilized. The solid was dissolved in water (20.0 mL) and (NH₄)HCO₃ (1.21 g, 15.3 mmol) was added at 22 °C. The mixture was stirred at 22 °C for 1 h, was filtered, concentrated, and lyophilized. The solid was dissolved in water (20.0 mL) and (NH₄)₂SO₄ (313 mg, 2.37 mmol) was added at 22 °C. The mixture was stirred at 22 °C for 1 h and was filtered on medium frit. The filtrate was added dropwise to MeOH (400 mL) and was stirred at 22 °C for 1 h. The suspension was centrifuged (rotation speed: 1765 rpm) for 30 min. MeOH supernatant was carefully removed, and the solid was dissolved in distilled water (15.0 mL). The solution was evaporated under reduced pressure to remove residual MeOH and lyophilized to provide the title compound (Compound 4) as a solid (135 mg, 50%). ¹H NMR (400 MHz, D₂O) δ 5.91 (d, J = 3.7 Hz, 1H), 5.42 (d, J = 2.0 Hz, 1H), 5.32 (d, J = 1.9 Hz, 1H), 4.57 (dd, AttyDktNo. R1119431100WO J = 7.2, 4.7 Hz, 1H), 4.45 (dd, J = 4.7, 2.0 Hz, 1H), 4.42 – 4.39 (m, 1H), 4.36 (d, J = 6.1 Hz, 1H), 4.29 – 4.19 (m, 3H), 4.03 – 3.81 (m, 7H), 3.81 – 3.67 (m, 4H), 3.62 (dt, J = 3.2, 1.3 Hz, 1H), 3.52 – 3.33 (m, 3H), 3.26 – 3.11 (m, 2H), 2.37 – 2.25 (m, 1H), 2.25 – 2.16 (m, 1H), 2.15 – 2.00 (m, 2H), 1.46 (d, J = 6.8 Hz, 3H). Example 7. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3- amino-6-((R)-1-aminoethyl)-5-hydroxytetrahydro-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 5) OH OH H H₂N N NH_{2 2} (Compound 5) Step A. Synthesis of -6-(p-tolylthio)tetrahydro-2H-pyran-

2-yl)methylene)-2-methylpropane-2-sulfinamide [00372] Anhydrous CuSO₄ (66.3 g, 416 mmol) was added to a mixture of (2S,3S,5R)-5-azido-3-benzyloxy-6- (p-tolylsulfanyl)tetrahydropyran-2-carbaldehyde (68.4 g, 175 mmol) and (R)-2-methylpropane-2-sulfinamide (63.6 g, 524 mmol) in dry DCM (667 mL) at 22 °C. The mixture was stirred at 22 °C for 48 h. The mixture was diluted with DCM (300 mL), filtered through Celite, washed with DCM (2 x 100 mL), and the filtrate was concentrated. The residue was diluted with water (500 mL), sat. aq. NH₄Cl (250 mL) and EtOAc (300 mL). The aqueous phase was extracted with EtOAc (3 x 200 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (200 mL), sat. aq. NaHCO₃ (200 mL), brine (200 mL), dried (Na₂SO₄), filtered and concentrated. The residue was purified by silica gel chromatography (2 x 330 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-20%) to provide the title compound as an oil (55.0 g, 65%). m/z (ES+) [M+H]⁺ = 487.0; HPLC (A05) t_R = 1.84 min. Step B. Synthesis of (R)-N-((1R)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H- pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide [00373] MeMgBr (51.2 mL, 154 mmol, 3.00 mol/L in Et₂O) was added slowly to a mixture of (NE,R)-N- [[(2R,3S,5R,6S)-5-azido-3-benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methylene]-2-methyl- propane-2-sulfinamide (24.1 g, 49.5 mmol) in dry DCM (338 mL) under nitrogen at -78 °C. The mixture was stirred at -78 °C for 2 h, at -60 °C for 7 h and was diluted with sat. aq. NH₄Cl (50.0 mL). The aqueous phase was extracted with EtOAc (3 x 50.0 mL), and the combined organic extracts were washed with brine (100 mL), dried (Na₂SO₄), filtered, concentrated. The residue was purified by silica gel chromatography (330 g AttyDktNo. R1119431100WO high-capacity cartridge, liquid loading PhMe) using a gradient of EtOAc in hexanes (0-20%) to provide the title compound as an oil (9.80 g, 39%). m/z (ES+) [M+H]⁺ = 503.2; HPLC (A05) t_R = 1.84 min. Step C. Synthesis of (R)-N-((1R)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H- pyran-2-yl)ethyl)-N-benzyl-2-methylpropane-2-sulfinamide [00374] KOtBu (8.79 g, 78.3 mmol) was added to a mixture of (R)-N-[(1R)-1-[(2R,3S,5R,6S)-5-azido-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (17.5 g, 34.8 mmol) and BnBr (11.4 mL, 95.7 mmol) in dry DMF (148 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h, and at 22 °C for 48 h. A second portion of BnBr (4.13 mL, 34.8 mmol) and KOtBu (3.91 g, 34.8 mmol) was added to the mixture at 22 °C. The mixture was stirred at 22 °C for 1 h. The mixture was diluted with AcOH (7.18 mL, 125 mmol), EtOAc (250 mL), water (250 mL), and sat. aq. NH₄Cl (500 mL) at 22 °C. The aqueous phase was extracted with EtOAc (3 x 100 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (100 mL), brine (100 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (220 g cartridge, gold) using a gradient of EtOAc in hexanes (0-25%) to provide the title compound as an oil (14.4 g, 70%). m/z (ES+) [M+Na]⁺ = 615.2; HPLC (A05) t_R = 1.95 min. Step D. Synthesis of benzyl ((1R)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H- pyran-2-yl)ethyl)(benzyl)carbamate [00375] HCl (1.0N, 38.5 mL) was added to a mixture of (R)-N-[(1R)-1-[(2R,3S,5R)-5-azido-3-benzyloxy-6- (p-tolylsulfanyl)tetrahydropyran-2-yl]ethyl]-N-benzyl-2-methyl-propane-2-sulfinamide (16.3 g, 27.5 mmol) in dioxane (143 mL) at 22 °C. The mixture was stirred at 22 °C for 30 min to provide the free amine. m/z (ES+) [M+H]⁺ = 489.2; HPLC (A05) t_R = 1.81 min. The mixture was diluted with PBS buffer 10X (176 mL) and CbzCl (7.82 mL, 55.0 mmol) was added dropwise. The mixture was stirred at 22 °C for 2 h and was diluted with EtOAc (200 mL). The aqueous phase was extracted with EtOAc (3 x 200 mL), and the combined organic extracts were washed with brine (50.0 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (220 g cartridge) using a gradient of EtOAc in hexanes (0-12%) to provide the title compound as a solid (13.5 g, 79%). m/z (ES+) [M+H]⁺ = 623.4; HPLC (A05) t_R = 2.15 min. Step E. Synthesis of benzyl ((1R)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-6-hydroxytetrahydro-2H-pyran- 2-yl)ethyl)(benzyl)carbamate [00376] NBS (4.03 g, 22.7 mmol) was added to a solution of benzyl N-[(1R)-1-[(2R,3S,5R,6R)-5-azido-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (12.3 g, 19.7 mmol) in acetone (480 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h and was diluted with 1:1 sat. aq. NaHCO₃/Na₂S₂O₃ (300 mL). The aqueous layer was extracted with EtOAc (2 x 100 mL), and the combined organic extracts were dried (Na₂SO₄), filtered and concentrated. The residue was purified by silica gel chromatography (200 g cartridge, dry loading) using a gradient of EtOAc in hexanes (0-40%) to provide the title compound as a solid (8.40 g, 83%). m/z (ES+) [M+H]⁺ = 517.2; HPLC (A05) t_R = 1.77 and 1.80 min. Step F. Synthesis of (3R,5S,6R)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)-5- (benzyloxy)tetrahydro-2H-pyran-2-yl (E)-2,2,2-trifluoro-N-phenylacetimidate AttyDktNo. R1119431100WO [00377] Cs₂CO₃ (10.6 g, 32.5 mmol) was added to a mixture of 2,2,2-trifluoro-N-phenyl-acetimidoyl chloride (4.67 mL, 29.3 mmol) and benzyl N-[(1R)-1-[(2R,3S,5R)-5-azido-3-benzyloxy-6-hydroxy-tetrahydropyran-2- yl]ethyl]-N-benzyl-carbamate (8.40 g, 16.3 mmol) in dry DCM (89.7 mL) at 22 °C under nitrogen. The mixture was stirred at 22 °C for 1 h 30, was filtered, washed with DCM (3 x 50.0 mL), and the filtrate was concentrated. The residue was purified by silica gel chromatography (200 g cartridge, dry pack) using a gradient of EtOAc in hexanes (0-40%) to provide the title compound as an oil (10.7 g, 96%). m/z (ES+) [M+Na]⁺ = 710.3; HPLC (A05) t_R = 2.04 min. Step G. Synthesis of dibenzyl ((1R,2R,3S,4R,5R,6S)-4-(((3R,5S,6R)-3-azido-6-((R)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-2,5,6- trihydroxycyclohexane-1,3-diyl)dicarbamate [00378] (3R,5S,6R)-3-Azido-6-[(1R)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]-5-benzyloxy- tetrahydropyran-2-yl] (1E)-2,2,2-trifluoro-N-phenyl-ethanimidate (10.7 g, 15.6 mmol) and [(1S,2R,3S,4S,5R,6R)-3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl] acetate (13.4 g, 23.3 mmol) were co-evaporated with dry toluene (2 x 50.0 mL), charged with pre-activated 4Å molecular sieves (11.0 g) and dried under vacuum for 30 min. The residue was dissolved in dry 2-MeTHF (310 mL) under nitrogen. Bi(OTf)₃ (11.2 g, 17.1 mmol) was added to the mixture at 0 °C. The mixture was stirred at 0 °C for 1 h and at 21 °C for 16 h. The mixture was filtered, and the solids were washed with DCM (2 x 50.0 mL). The filtrate was diluted with sat. aq. NH₄Cl (150 mL). The aqueous phase was extracted with DCM (3 x 100 mL), and the combined organic extracts were washed brine (200 mL), dried (Na₂SO₄), filtered and concentrated. m/z (ES+) [M+H]⁺ = 1071.5; HPLC (A05) t_R = 1.94 min. The residue was dissolved in MeOH (225 mL) and NaOMe (4.60 M in MeOH, 2.71 mL, 12.4 mmol) was added at 22 °C. The mixture was stirred at 22 °C for 2 h 30 and additional NaOMe (4.60 mol/L in MeOH, 0.700 mL, 3.22 mmol) was added. The mixture was stirred at 22 °C for 16 h, and was diluted with AcOH (2.67 mL, 46.7 mmol). The mixture was filtered, and the solids were triturated in EtOAc (3 x 40.0 mL). The filtrate was concentrated, co-evaporated with toluene (30.0 mL). The residue was purified by silica gel chromatography (200 g cartridge, gold, dry pack) using a gradient of EtOAc in hexanes (0-80%) to provide a mixture of anomers of the title compound as a solid (3.24 g, 22%, over 2 steps). m/z (ES+) [M+H]⁺ = 945.4; HPLC (A05) t_R = 5.54 and 5.60 min. Step H. Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((3R,5S,6R)-3-azido-6-((R)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-6- hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate [00379] PPTS (356 mg, 1.42 mmol) was added to a mixture of benzyl N-[(1R)-1-[(2R,3S,5R,6S)-5-azido-3- benzyloxy-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy- cyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (3.24 g, 3.43 mmol) and 1,1- dimethoxycyclohexane (4.42 mL, 29.1 mmol) in dry DME (16.4 mL) at 22 °C under nitrogen. The mixture was stirred at 90 °C for 16 h under nitrogen, cooled to 22 °C, and concentrated. The residue was purified by silica gel chromatography (100 g cartridge, gold, PhMe loading) using a gradient of EtOAc in hexanes (0- AttyDktNo. R1119431100WO 40%) to provide the title compound as a solid (1.62 g, 46%). m/z (ES+) [M+H]⁺ = 1025.7; HPLC (A05) t_R = 2.03 min. Step I. Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((3R,5S,6R)-3-azido-6-((R)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-6- (benzyloxy)hexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate [00380] NaH (0.289 g, 7.21 mmol, 60 % in mineral oil) was added to a mixture of benzyl N-[(1R)-1- [(2R,3S,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-3-benzyloxy-tetrahydropyran-2-yl]ethyl]- N-benzyl-carbamate (1.72 g, 1.68 mmol) and BnBr (0.598 mL, 5.03 mmol) in dry THF (38.5 mL) at 0 °C under nitrogen. The mixture was stirred at 22 °C for 1 h 30. The mixture was diluted with AcOH (0.432 mL, 7.55 mmol) and was concentrated. The residue was purified by silica gel chromatography (40 g cartridge, gold, dry loading) using a gradient of EtOAc in hexanes (0-40%) to provide the title compound as a solid (1.50 g, 80%). m/z (ES+) [M+H]⁺ = 1115.4; HPLC (A05) t_R = 2.18 min. Step J. Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((3R,5S,6R)-3-azido-6-((R)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)tetrahydro-2H-pyran-2-yl)oxy)-2- (benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate [00381] A mixture of benzyl N-[(1R)-1-[(2R,3S,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-3-benzyloxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (1.50 g, 1.34 mmol) in THF (6.09 mL), water (2.51 mL) and AcOH (4.03 mL) was stirred at 80 °C for 24 h. The mixture was cooled to 22 °C, concentrated and was diluted with EtOAc (50.0 mL). The pH was adjusted to pH ^ 8 with sat. aq. NaHCO₃. The aqueous phase was extracted with EtOAc (2 x 20.0 mL), and the combined organic extracts were washed with brine (50.0 mL), dried (Na₂SO₄), filtered, and concentrated to provide the title compound as a solid (1.40 g, quant.). m/z (ES+) [M+H]⁺ = 1035.6; HPLC (A05) t_R = 1.93 min. Step K. Synthesis of benzyl ((1R)-1-((2R,3S,5R)-5-azido-3-(benzyloxy)-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 [00382] NaOtBu (650 mg, 6.76 mmol) was added to a mixture of benzyl N-[(1R)-1-[(2R,3S,5R,6S)-5-azido- 3-benzyloxy-6-[(1R,2S,3R,4S,5S,6R)-3-benzyloxy-2,4-bis(benzyloxycarbonylamino)-5,6-dihydroxy- cyclohexoxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (1.40 g, 1.35 mmol) and TEBA-Cl (1.54 g, 6.76 mmol) in dry DMF (22.9 mL) at 0 °C. The mixture was stirred at 0 °C for 30 min and diluted with brine (300 mL). The solid was filtered and triturated with EtOAc (2 x 50.0 mL). The aqueous phase was extracted with EtOAc (3 x 40.0 mL), and the combined organic extracts were washed with brine (200 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (40 g cartridge, gold, dry loading) using a gradient of EtOAc in hexanes (0-65%) to provide the title compound as a solid (819 mg, 65%). m/z (ES+) [M+H]⁺ = 927.4, HPLC (A05) t_R = 1.87 min. AttyDktNo. R1119431100WO Step L. 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,5S,6R)-3-azido-6-((R)-1- (benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)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 [00383] Potassium carbonate (0.733 g, 5.30 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 (1.21 g, 2.28 mmol) and CCl₃CN (0.532 mL, 5.30 mmol) in dry DCM (21.3 mL) under nitrogen. The mixture was stirred at 22 °C for 18 h and was filtered. The solids were rinsed with DCM (3 x 20.0 mL) and concentrated to provide an oil. The and benzyl N-[(1R)-1-[(2R,3S,5R,6S)-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-3-benzyloxy-tetrahydropyran-2-yl]ethyl]-N-benzyl- carbamate (819 mg, 0.883 mmol) were co-evaporated with dry toluene (3 x 20.0 mL) under reduced pressure and pre-activated 4Å molecular sieves (1.0 g) were added. The mixture was dissolved in dry DCM (21.3 mL) under nitrogen. The mixture was cooled to -78 °C and BF₃ ^Et₂O (1.65 mL, 13.3 mmol) was added. The mixture was stirred at 22 °C for 1 h 20 and was diluted with sat. aq. NaHCO₃ (50.0 mL). The mixture was filtered, and the solids were washed with DCM (3 x 20.0 mL). The aqueous layer was extracted with DCM (3 x 20 mL), and the combined organic extracts were washed with brine (30.0 mL), dried (Na2SO4), filtered, concentrated. The residue was purified by silica gel chromatography (50 g cartridge, gold, dry pack) using a gradient of EtOAc in hexanes (0-65%) to provide the title compound as a solid (1.21 g, 95%). m/z (ES+) [M- CO]⁺ = 1411.7; HPLC (A05) t_R= 1.96 min. Step L. Synthesis of benzyl ((R)-1-((2R,3S,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-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy- 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3- hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate [00384] Lithium hydroxide monohydrate (1.06 g, 25.2 mmol) was added to a mixture 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,5S,6R)-3-azido-6-((R)- 1-(benzyl((benzyloxy)carbonyl)amino)ethyl)-5-(benzyloxy)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 (1.21 g, 0.841 mmol) in dioxane (22.7 mL) and water (11.4 mL) at 22 °C under nitrogen. The mixture was stirred at 40 °C for 18 h and dioxane was removed under reduced pressure. EtOAc (20.0 mL) and H₂O (50.0 mL) were added, and the aqueous phase was extracted with EtOAc (3 x 50.0 mL), and the combined organic extracts were washed with brine (50.0 mL), dried (MgSO₄), filtered, and concentrated to provide the title compound as a solid (842 mg, 80%). m/z (ES+) [M+H]⁺ = 1245.7; HPLC (A05) t_R = 1.76 min. Step M. Synthesis of benzyl ((R)-1-((2R,3S,5R,6R)-5-azido-6-(((1R,2R,3S,4S,5R,6S)-4-((2R,3R)-4-azido- 3-fluoro-2-((4-methoxybenzyl)oxy)butanamido)-2-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-azido-6- AttyDktNo. R1119431100WO (azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3- hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate [00385] DMTMM chloride (64.5 mg, 0.233 mmol) was added to a mixture of benzyl ((R)-1-((2R,3S,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- dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5- (benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro- 2H-pyran-2-yl)ethyl)(benzyl)carbamate (242 mg, 0.194 mmol) and (2R,3R)-4-azido-3-fluoro-2-[(4- methoxyphenyl)methoxy]butanoic acid (60.6 mg, 0.214 mmol) in dry THF (4.43 mL). The mixture was stirred at 22 °C for 48 h, and (2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoic acid (33.0 mg, 0.117 mmol) and DMTMM chloride (32.3 mg, 0.117 mmol) were added. The mixture was stirred at 22 °C for 5 h and diluted with sat. aq. NH₄Cl (5.00 mL) and EtOAc (5.00 mL). The aqueous phase was extracted with EtOAc (3 x 5.00 mL), and the combined organic extracts were washed with brine (5.00 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (12 g cartridge, gold, dry pack) using a gradient of MeOH in DCM (0-3%) to provide the title compound as a solid (178 mg, 61%). m/z (ES+) [M-CD rings+H]⁺ = 1166.7; HPLC (A05) t_R = 1.92 min. Step N. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3-amino-6- ((R)-1-aminoethyl)-5-hydroxytetrahydro-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 5) [00386] A mixture of benzyl ((R)-1-((2R,3S,5R,6R)-5-azido-6-(((1R,2R,3S,4S,5R,6S)-4-((2R,3R)-4-azido-3- fluoro-2-((4-methoxybenzyl)oxy)butanamido)-2-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-azido-6- (azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran- 2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-3- (benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate intermediate (178 mg, 0.118 mmol) in MeOH (2.22 mL) was added to a suspension of Pd(OH)₂ (0.736 g, 1.05 mmol, 20% wet) in AcOH (1.83 mL) and water (0.721 mL) at 22 °C under nitrogen. The mixture was evacuated with hydrogen three times, and stirred at 22 °C under continuous bubbling of hydrogen balloon pressure for 3.5 h. The mixture was evacuated with nitrogen, bubbled with nitrogen for 5 min. The suspension was filtered on Celite, washing with water (100 mL), MeOH (100 mL), water (2 x 50.0 mL). The filtrate was concentrated (rotavapor bath temperature < 35 °C) and was purified by preparative HPLC using 5% of MeCN in water ^10 mM Ammonium bicarbonate pH 10 ^, and was lyophilized. The solid was dissolved in water (10.0 mL) and (NH₄)₂SO₄ (46.7 mg, 0.354 mmol) was added. The mixture was stirred at 22 °C for 5 min, filtered on a medium frit and added dropwise to MeOH (400 mL) under vigorous stirring. The suspension was centrifuged (rotation speed: 1765 rpm) for 1 h. MeOH supernatant was carefully removed, and the solid was dissolved in distilled water (25.0 mL). The solution was was evaporated under reduced pressure to remove residual MeOH and lyophilized to provide the AttyDktNo. R1119431100WO title compound (Compound 5) as a solid (46.0 mg, 38%, 99% purity by NMR).¹H NMR (400 MHz, D₂O) δ 6.08 (d, J = 3.9 Hz, 1H), 5.45 (d, J = 1.9 Hz, 1H), 5.41 – 5.21 (m, 2H), 4.59 – 4.33 (m, 4H), 4.26 (q, J = 5.2 Hz, 3H), 4.04 – 3.81 (m, 8H), 3.81 – 3.53 (m, 6H), 3.52 – 3.28 (m, 5H), 2.35 – 2.26 (m, 1H), 2.09 (q, J = 11.8 Hz, 1H), 1.34 (d, J = 6.8 Hz, 3H). Note: Missing hydrogens due to exchange with solvent. Example 8. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3- amino-6-((R)-1-aminoethyl)-5-hydroxytetrahydro-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- tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2- hydroxybutanamide (Compound 6) OH OH H H₂N N NH 2 (Compound 6) Step A. Synthesis of

-2-(((2S,3R,4S,5R)-4- (((2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy- 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-4-((S)-4- (((benzyloxy)carbonyl)amino)-2-hydroxybutanamido)-3-hydroxycyclohexyl)oxy)-3- (benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate [00387] Benzyl ((R)-1-((2R,3S,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-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3- hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (600 mg, 0.482 mmol), PyBOP (0.426 g, 0.819 mmol), DIPEA (0.206 mL, 1.20 mmol) and (2S)-4- (benzyloxycarbonylamino)-2-hydroxy-butanoic acid (183 mg, 0.723 mmol) were dissolved in dry DMF (9.30 mL) at 22 °C. The mixture was stirred at 22 °C for 25 min and was diluted with sat. aq. NH₄Cl (50.0 mL) and EtOAc (10.0 mL). The aqueous phase was extracted with EtOAc (4 x 10.0 mL), and the combined organic extracts were washed with brine (30.0 mL), dried (Na₂SO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (25 g high-capacity cartridge, dry loading) using a gradient of MeOH in DCM (0-7%) to provide the title compound as a solid (411 mg, 58%). m/z (ES+) [M-CD rings +H]⁺ = 1136.6; HPLC (A05) t_R = 1.84 min. AttyDktNo. R1119431100WO Step B. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3-amino-6-((R)-1- aminoethyl)-5-hydroxytetrahydro-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) [00388] Benzyl ((R)-1-((2R,3S,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-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-4-((S)-4- (((benzyloxy)carbonyl)amino)-2-hydroxybutanamido)-3-hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro- 2H-pyran-2-yl)ethyl)(benzyl)carbamate (Step A; 411 mg, 0.278 mmol) in MeOH (5.24 mL), water (1.70 mL) and AcOH (4.31 mL) was added to Pearlman's catalyst (0.585 g, 0.833 mmol, 20% purity) under nitrogen. The mixture was bubbled with hydrogen for 5 min and stirred at 22 °C under continuous bubbling hydrogen balloon pressure for 4 h. The mixture was evacuated with nitrogen, bubbled with nitrogen for 5 min. The suspension was filtered on Celite, washing with water (100 mL), MeOH (100 mL) and water (2 x 50.0 mL). The volatiles were evaporated under reduced pressure (rotavapor bath temperature < 35 °C). The material was purified by preparative HPLC with 5% of MeCN water ^10 mM ammonium bicarbonate pH 10 ^, and lyophilized. The solid was dissolved in water (10.0 mL) and (NH₄)₂SO₄ (110 mg, 0.833 mmol) was added. The mixture was stirred at 22 °C for 5 min, then filtered on a medium frit and added dropwise to MeOH (400 mL) under vigorous stirring. The suspension was centrifuged (rotation speed: 1765 rpm) for 2 x 30 min. MeOH supernatant was carefully removed, and the solid was dissolved in distilled water (25.0 mL). The solution was evaporated under reduced pressure to remove residual MeOH and lyophilized to provide the title compound (Compound 6) as a solid (172 mg, 61%, 86.6% purity by qNMR). m/z (ES+) [M+H]⁺ = 730.4; HPLC (A05) t_R = 0.12 min. ¹H NMR (400 MHz, D₂O) δ 6.08 (d, J = 3.8 Hz, 1H), 5.45 (d, J = 2.0 Hz, 1H), 5.32 (d, J = 1.8 Hz, 1H), 4.55 (dd, J = 7.0, 4.7 Hz, 1H), 4.46 (dd, J = 4.8, 2.0 Hz, 1H), 4.43 – 4.34 (m, 2H), 4.30 – 4.13 (m, 3H), 4.05 – 3.59 (m, 12H), 3.50 – 3.30 (m, 3H), 3.26 – 3.13 (m, 2H), 2.34 – 2.26 (m, 1H), 2.26 – 2.17 (m, 1H), 2.16 – 2.00 (m, 2H), 1.34 (d, J = 6.8 Hz, 3H). Note: 21 protons are missing due to deuterium exchange.

AttyDktNo. R1119431100WO Example 9. Synthesis of ((2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)- 3-amino-5-hydroxy-6-((methylamino)methyl)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) OH OH H H₂N N NH_{2 2} (Compound 7) Step A. Synthesis of (p-tolylthio)tetrahydro-2H-pyran-2- yl)methyl)-2-

[00389] NaBH₄ (2.30 g, 59.5 mmol) was added portion wise to a mixture of (NE,R)-N-[[(2R,3S,5R)-5-azido- 3-benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methylene]-2-methyl-propane-2-sulfinamide (21.0 g, 39.7 mmol) in dry MeOH (322 mL) at 0 °C. The mixture was stirred at 0 °C for 30 min and was diluted with sat. aq. NH₄Cl (200 mL) and EtOAc (300 mL). The aqueous phase was extracted with EtOAc (3 x 200 mL), and the combined organic extracts were washed with sat. aq. NH₄Cl (200 mL), sat. aq. NaHCO₃ (200 mL), brine (200 mL), dried (Na2SO4), filtered and concentrated. The residue was purified by silica gel chromatography (220 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-100%) to provide the title compound (R)-N-(((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H-pyran-2- yl)methyl)-2-methylpropane-2-sulfinamide as a solid (16.3 g, 84%). ¹H NMR (400 MHz, CDCl₃) δ 7.43 – 7.38 (m, 2H), 7.34 – 7.23 (m, 5H), 7.12 – 7.06 (m, 2H), 4.55 (d, J = 11.4 Hz, 1H), 4.45 – 4.33 (m, 2H), 3.56 (d, J = 12.3 Hz, 2H), 3.50 – 3.44 (m, 1H), 3.33 (ddd, J = 10.9, 9.2, 4.4 Hz, 1H), 3.26 – 3.18 (m, 2H), 2.59 (dt, J = 12.4, 4.7 Hz, 1H), 2.30 (s, 3H), 1.51 (td, J = 12.2, 11.0 Hz, 1H), 1.13 (s, 9H); m/z (ES+) [M+H]⁺ = 489.2; HPLC (A05) t_R = 1.81 min. Step B. Synthesis of (R)-N-(((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H-pyran-2- yl)methyl)-N,2-dimethylpropane-2-sulfinamide [00390] NaH (1.80 g, 47.0 mmol, 60% dispersion in mineral oil) was added to a mixture of (R)-N- [[(2R,3S,5R)-5-azido-3-benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methyl]-2-methyl-propane-2- sulfinamide (18.0 g, 36.8 mmol) in dry THF (208 mL) at 0 °C. The mixture was stirred at 0 °C for 20 min and MeI (3.44 mL, 55.3 mmol) was added. The mixture was stirred at 22 °C for 24 h and diluted with sat. aq. NH₄Cl (100.0 mL) and EtOAc (300.0 mL). The aqueous phase was extracted with EtOAc (3 x 300 mL), and the combined organic extracts were washed with brine (200 mL), dried (Na₂SO₄), filtered and concentrated. The residue was purified by silica gel chromatography (220 g high-capacity cartridge) using a gradient of AttyDktNo. R1119431100WO EtOAc in hexanes (0-100%) to provide the title compound as an oil (15.8 g, 85%). m/z (ES+) [M+H]⁺ = 503.2; HPLC (A05) t_R = 1.89 min. Step C. Synthesis of benzyl (((2R,3S,5R)-5-azido-3-(benzyloxy)-6-(p-tolylthio)tetrahydro-2H-pyran-2- yl)methyl)(methyl)carbamate [00391] HCl (44.0 mL, 44.0 mmol, 1 N in water) was added to a mixture of (R)-N-[[(2R,3S,5R)-5-azido-3- benzyloxy-6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methyl]-N,2-dimethyl-propane-2-sulfinamide (15.8 g, 31.4 mmol) in 1,4-dioxane (164 mL) at 22 °C. The mixture was stirred at 22 °C for 30 min. m/z (ES+) [M+H]⁺ = 399.1; HPLC (A05) t_R = 1.55 min. [00392] The pH of the mixture was adjusted by adding PBS buffer (201 mL) with pH probe monitoring (crude pH remaining around 7.6), and CbzCl (8.94 mL, 62.9 mmol) was added. The mixture was stirred at 22 °C for 2 h, then was diluted with EtOAc (250.0 mL). The aqueous phase was extracted with EtOAc (3 x 250.0 mL), and the combined organics extracts were washed with brine (250 mL), dried (MgSO₄), filtered and concentrated. The residue was purified by silica gel chromatography (220 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-100%) to provide the title compound as an oil (16.4 g, 92%). m/z (ES+) [M+H]⁺ = 533.2; HPLC (A05) t_R = 1.98 min. Step D. Synthesis of benzyl (((2R,3S,5R)-5-azido-3-(benzyloxy)-6-hydroxytetrahydro-2H-pyran-2- yl)methyl)(methyl)carbamate [00393] NBS (6.72 g, 37.8 mmol) was added to a mixture of benzyl N-[[(2R,3S,5R,6R)-5-azido-3-benzyloxy- 6-(p-tolylsulfanyl)tetrahydropyran-2-yl]methyl]-N-methyl-carbamate (17.5 g, 32.9 mmol) in acetone (801 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h and was diluted with 1:1 sat. NaHCO₃/Na₂S₂O₃ (300.0 mL). The aqueous phase was extracted with EtOAc (3 x 250 mL), and the combined organic extracts were dried (MgSO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (220 g high- capacity cartridge) using a gradient of EtOAc in hexanes (0-100%) to provide the title compound as an oil (13.5 g, 92%). m/z (ES+) [M+H]⁺ = 427.2; HPLC (A05) t_R= 1.70 and 1.72 min (mixture of anomers). Step E. Synthesis of (3R,5S,6R)-3-azido-5-(benzyloxy)-6- ((((benzyloxy)carbonyl)(methyl)amino)methyl)tetrahydro-2H-pyran-2-yl (E)-2,2,2-trifluoro-N- phenylacetimidate [00394] Cs₂CO₃ (21.2 g, 65.2 mmol) was added to a mixture of 2,2,2-trifluoro-N-phenyl-acetimidoyl chloride (9.37 mL, 58.7 mmol) and benzyl N-[[(2R,3S,5R)-5-azido-3-benzyloxy-6-hydroxy-tetrahydropyran-2- yl]methyl]-N-methyl-carbamate (13.9 g, 32.6 mmol) in dry DCM (180 mL) at 22 °C under nitrogen. The mixture was stirred at 22 ^oC for 2 h, then filtered on Celite, washing with DCM (3 x 50.0 mL) and concentrated. The residue was purified by silica gel chromatography (220 g high-capacity cartridge, dry pack) using a gradient of EtOAc in hexanes (0-50%) to provide the title compound as an oil (16.1 g, 83%). m/z (ES+) [M+Na]⁺ = 620.1; HPLC (A05) t_R = 2.00 min. Step F. Synthesis of dibenzyl ((1R,2R,3S,4R,5R,6S)-4-(((3R,5S,6R)-3-azido-5-(benzyloxy)-6- ((((benzyloxy)carbonyl)(methyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2,5,6- trihydroxycyclohexane-1,3-diyl)dicarbamate AttyDktNo. R1119431100WO [00395] [(3R,5S,6R)-3-Azido-5-benzyloxy-6[[benzyloxycarbonyl(methyl)amino]methyl]tetrahydropyran-2- yl] (1E)-2,2,2-trifluoro-N-phenyl-ethanimidate (16.1 g, 26.9 mmol) and [(1S,2R,3S,4S,5R,6R)-3,4-diacetoxy- 2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl] acetate (23.1 g, 40.4 mmol) were co-evaporated with dry toluene (2 x 50.0 mL) at 25 ^oC. The mixture was dried under vacuum for 30 min. The residue was dissolved with anhydrous 2-MeTHF (412 mL) and transferred via a canula to a RBF charged with pre- activated 4Å molecular sieves (16.0 g) at 0 °C, and Bi(OTf)₃ (19.4 g, 29.6 mmol) was added in one portion at 0 °C. The mixture was stirred at 0 °C for 1 h and at 22 °C for 16 h. The mixture was filtered through Celite and the filtrate was diluted with sat. aq. NH₄Cl (200 mL). The aqueous phase was extracted with DCM (3 x 250 mL), and the combined organic extracts were washed brine (150 mL), dried (MgSO₄), filtered and concentrated. The residue was used without further purification to the next step (54.2 g crude). m/z (ES+) [M+H]⁺ = 981.4; HPLC (A05) tR = 5.85 min (inseparable mixture of diastereomers). [00396] NaOMe (9.61 mL, 44.2 mmol, 4.60 M in MeOH) was added to the crude mixture from the previous step in MeOH (800 mL) at 22 ^oC. The mixture was stirred at 22 ^oC for 1 h 30, diluted with AcOH (9.48 mL, 166 mmol) until pH ~5, and was concentrated. The solid was filtered, washed with EtOAc (500 mL), MeOH (500 mL) and the filtrate was concentrated. The residue was purified by silica gel chromatography (350 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-100%) to provide the title compound as a solid (7.80 g, 34% over 2 steps, mixture of diastereomers). m/z (ES+) [M+H]⁺ = 855.4, HPLC (A05) t_R= 5.89 min. Step G. Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((3R,5S,6R)-3-azido-5-(benzyloxy)-6- ((((benzyloxy)carbonyl)(methyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6- hydroxyhexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate [00397] PPTS (292 mg, 1.16 mmol) was added to a mixture of benzyl N-[[(2R,3S,5R,6R)-5-azido-3- benzyloxy-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy- cyclohexoxy]tetrahydropyran-2-yl]methyl]-N-methyl-carbamate (2.40 g, 2.81 mmol) and 1,1- dimethoxycyclohexane (3.62 mL, 23.8 mmol) in dry DME (13.4 mL) at 22 °C. The mixture was stirred at 90 °C under nitrogen for 2 h and was concentrated. The residue was purified by silica gel chromatography (40 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-60%) to provide the title compound as a solid (2.10 g, 80%). m/z (ES+) [M+H]⁺ = 936.5; HPLC (A05) t_R = 2.00 min. Step H. Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((3R,5S,6R)-3-azido-5-(benzyloxy)-6- ((((benzyloxy)carbonyl)(methyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6- (benzyloxy)hexahydrospiro[benzo[d][1,3]dioxole-2,1'-cyclohexane]-5,7-diyl)dicarbamate [00398] NaH (386 mg, 9.66 mmol, 60% dispersion in mineral oil) was added to a mixture of benzyl N- [[(2R,3S,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-3-benzyloxy-tetrahydropyran-2- yl]methyl]-N-methyl-carbamate (2.10 g, 2.25 mmol) and BnBr (0.800 mL, 6.74 mmol) in dry THF (51.6 mL) at 0 °C under nitrogen. The mixture was stirred at 22 °C for 3 h. The mixture was diluted with AcOH (0.579 mL, 10.1 mmol) and EtOAc (20.0 mL). Silica gel was added to the mixture and was concentrated. The residue AttyDktNo. R1119431100WO was purified by silica gel chromatography (40 g high-capacity cartridge) using a gradient of EtOAc in hexanes (0-40%) to provide the title compound as a solid (2.07 mg, 90%). m/z (ES+) [M+H]⁺ = 1026.6; HPLC (A05) t_R = 2.16 min. Step I. Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((3R,5S,6R)-3-azido-5-(benzyloxy)-6- ((((benzyloxy)carbonyl)(methyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6- dihydroxycyclohexane-1,3-diyl)dicarbamate [00399] AcOH (7.31 mL) was added to a mixture of benzyl N-[[(2R,3S,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-3-benzyloxy-tetrahydropyran-2-yl]methyl]-N-methyl-carbamate (2.50 g, 2.44 mmol) in THF (11.0 mL) and water (4.55 mL) at 22 °C. The mixture was stirred at 80 °C for 24 h and cooled to 22 °C. The mixture was concentrated, diluted with EtOAc (200 mL) and the combined organic extracts were washed with sat. aq. NaHCO₃ (2 x 100 mL), brine (100 mL), dried (MgSO₄), filtered, and concentrated to provide the title compound as a solid (2.19 g, 95%). m/z (ES+) [M+H]⁺ = 946.4; HPLC (A05) t_R = 1.83 min. Step J. Synthesis of benzyl (((2R,3S,5R)-5-azido-3-(benzyloxy)-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)(methyl)carbamate [00400] NaOtBu (1.11 g, 11.6 mmol) was added to a mixture of benzyl N-[[(2R,3S,5R,6R)-5-azido-3- benzyloxy-6-[(1R,2S,3R,4S,5S,6R)-3-benzyloxy-2,4-bis(benzyloxycarbonylamino)-5,6-dihydroxy- cyclohexoxy]tetrahydropyran-2-yl]methyl]-N-methyl-carbamate (2.19 g, 2.32 mmol) and TEBA-Cl (2.64 g, 11.6 mmol) in dry DMF (39.2 mL) at 0 °C. The mixture was stirred at 0 °C for 30 min and diluted with brine (300.0 mL). The aqueous phase was extracted with EtOAc (3 x 100 mL), and the combined organic extracts were washed with brine (150 mL) and concentrated. The residue was purified by silica gel chromatography (40 g high-capacity cartridge, dry loading) using a gradient of EtOAc in hexanes (0-75%) to provide the title compound as a solid (1.55 g, 80% yield). m/z (ES+) [M+H]⁺ = 837.3; HPLC (A05) t_R = 1.84 min. Step K. 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,5S,6R)-3-azido-5-(benzyloxy)-6- ((((benzyloxy)carbonyl)(methyl)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 [00401] K₂CO₃ (1.42 g, 10.3 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 (2S,3R,4R,5R,6R)-6-(((2R,3R,4R)-4-acetoxy-2-(acetoxymethyl)-5-hydroxytetrahydrofuran-3-yl)oxy)- 5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (2.34 g, 4.41 mmol) and CCl₃CN (1.03 mL, 10.3 mmol) in dry DCM (41.1 mL). The mixture was stirred at 22 °C for 18 h, then filtered on a short pad of Celite, washing with DCM and concentrated to give a syrup. The syrup and benzyl N-[[(2R,3S,5R,6R)-6- [[(3aR,4S,5S,6R,7S,7aS)-4-benzyloxy-5-(benzyloxycarbonylamino)-7-hydroxy-2-oxo-3a,4,5,6,7,7a- AttyDktNo. R1119431100WO hexahydro-3H-1,3-benzoxazol-6-yl]oxy]-5-azido-3-benzyloxy-tetrahydropyran-2-yl]methyl]-N-methyl- carbamate (1.43 g, 1.71 mmol) were co-evaporated with dry toluene (3 x 20.0 mL) under reduced pressure. The mixture was dried under vacuum for 30 min and dissolved in dry DCM (41.1 mL). The mixture was transferred to an RBF charged with pre-activated 4Å molecular sieves (2.00 g). The mixture was cooled to - 78 °C, and BF₃ ^.OEt₂ (3.18 mL, 25.8 mmol) was added at -78 °C. The mixture was stirred at 22 °C for 90 min and diluted with sat. aq. NaHCO₃ (100 mL) and DCM (100 mL). The mixture was filtered through Celite, washing with DCM (3 x 50.0 mL). The aqueous phase was extracted with DCM (3 x 50.0 mL), and the combined organic extracts were washed with brine (100 mL), dried (MgSO₄), filtered, and concentrated. The residue was purified by silica gel column (40 g high-capacity cartridge, dry loading) using a gradient of EtOAc in hexanes (0-75%) to provide the title compound as a solid (1.95 g, 85%). m/z (ES+) [M+Na]⁺ = 1372.6; HPLC (A05) tR = 1.90 min. Step L. Synthesis of benzyl (((2R,3S,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-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy- 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3- hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl)(methyl)carbamate [00402] LiOH^.H₂O (1.82 g, 43.4 mmol) was added to a mixture 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,5S,6R)-3-azido-5-(benzyloxy)-6- ((((benzyloxy)carbonyl)(methyl)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 (1.95 g, 1.45 mmol) in dioxane (39.1 mL) and water (19.5 mL) at 22 °C under nitrogen. The mixture was stirred at 40 °C for 18 h and dioxane was removed under reduced pressure. The mixture was diluted with EtOAc (150 mL) and H₂O (50.0 mL). The aqueous phase was extracted with EtOAc (3 x 100 mL), and the combined organic extracts were washed with brine (50.0 mL), dried (MgSO₄), filtered, and concentrated to provide the title compound as a solid (1.50 g, 90%). m/z (ES+) [M+H]⁺ = 1156.6; HPLC (A05) t_R = 1.70 min. Step M. Synthesis of benzyl (((2R,3S,5R,6R)-5-azido-6-(((1R,2R,3S,4S,5R,6S)-4-((2R,3R)-4-azido-3- fluoro-2-((4-methoxybenzyl)oxy)butanamido)-2-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-azido-6- (azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3- hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro-2H-pyran-2-yl)methyl)(methyl)carbamate [00403] DMTMM chloride (124 mg, 0.450 mmol) was added to a solution of benzyl (((2R,3S,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- dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5- (benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro- 2H-pyran-2-yl)methyl)(methyl)carbamate (296 mg, 0.256 mmol) and (2R,3R)-4-azido-3-fluoro-2-[(4- methoxyphenyl)methoxy]butanoic acid (109 mg, 0.384 mmol) in dry THF (5.84 mL) at 22 °C. The mixture was stirred at 22 °C for 18 h, then was diluted with sat. aq. NH₄Cl (20.0 mL) and EtOAc (40.0 mL). The AttyDktNo. R1119431100WO aqueous phase was extracted with EtOAc (3 x 30.0 mL), and the combined organic extracts were washed with brine (2 x 10.0 mL), dried (MgSO₄), filtered and concentrated. The residue was purified by silica gel chromatography (24 g high-capacity cartridge) using a gradient of EtOAc in hexanes (50-100%) and using a gradient of EtOAc in MeOH (0-20%) to provide the title compound as a solid (239 mg, 66%). m/z (ES+) [M- CD rings+H]⁺ = 1049.5; HPLC (A05) t_R = 1.84 min. Step N. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3-amino-5- hydroxy-6-((methylamino)methyl)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) [00404] A mixture of benzyl (((2R,3S,5R,6R)-5-azido-6-(((1R,2R,3S,4S,5R,6S)-4-((2R,3R)-4-azido-3-fluoro- 2-((4-methoxybenzyl)oxy)butanamido)-2-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)- 4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5- (benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro-2H- pyran-2-yl)methyl)(methyl)carbamate (239 mg, 0.168 mmol) in MeOH (7.41 mL) was added to a suspension of Pd(OH)₂ (709 mg, 1.01 mmol, 20% wet) in AcOH (6.09 mL) and water (2.40 mL) at 22 °C under nitrogen. The mixture was evacuated with hydrogen three times and was stirred at 22 °C under continuous bubbling of hydrogen balloon pressure for 4 h. The mixture was evacuated with nitrogen, bubbled with nitrogen for 5 min. The suspension was filtered on Celite, washed with water (100 mL), MeOH (100 mL) and water (2 x 50.0 mL). The volatiles were evaporated under reduced pressure (rotavapor bath temperature < 35 °C). The residue was purified by preparative HPLC using 5% of MeCN in water ^10 mM Ammonium bicarbonate pH 10 ^ and was lyophilized to give a solid. The solid was dissolved in water (10.0 mL), and (NH₄)₂SO₄ (66.7 mg, 0.505 mmol) was added. The mixture was stirred at 22 °C for 15 min, then filtered on a medium frit and added dropwise to MeOH (500 mL) under vigorous stirring. The suspension was centrifuged (rotation speed: 1765 rpm) for 30 min. MeOH supernatant was carefully removed, and the solid was dissolved in distilled water (15.0 mL). The solution was evaporated under reduced pressure to remove residual MeOH and lyophilized to provide the title compound (Compound 7) as a solid (145 mg, 83%, >99% purity by q-NMR).¹H NMR (400 MHz, D₂O) δ 5.95 (d, J = 3.7 Hz, 1H), 5.42 (d, J = 1.8 Hz, 1H), 5.31 (d, J = 1.8 Hz, 1H), 4.59 – 4.43 (m, 3H), 4.38 – 4.22 (m, 4H), 4.07 – 3.80 (m, 7H), 3.72 (ddd, J = 22.7, 11.1, 6.8 Hz, 4H), 3.61 (dt, J = 3.0, 1.2 Hz, 1H), 3.57 – 3.34 (m, 6H), 3.26 (dd, J = 13.2, 8.3 Hz, 1H), 2.81 (s, 3H), 2.30 (dt, J = 11.8, 4.4 Hz, 1H), 2.10 (q, J = 11.8 Hz, 1H). m/z (ES+) [M+H]⁺ = 748.5; HPLC (A05) t_R = 0.116 min. AttyDktNo. R1119431100WO Example 10. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3- amino-5-hydroxy-6-((methylamino)methyl)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 8) OH OH H H₂N N NH_{2 2} (Compound 8) Step A. Synthesis

- -2-(((2S,3R,4S,5R)-4- (((2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy- 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-4-((S)-4- (((benzyloxy)carbonyl)amino)-2-hydroxybutanamido)-3-hydroxycyclohexyl)oxy)-3- (benzyloxy)tetrahydro-2H-pyran-2-yl)methyl)(methyl)carbamate [00405] PyBOP (501 mg, 0.962 mmol) was added to a mixture of benzyl (((2R,3S,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- dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5- (benzyloxy)-6-(((benzyloxy)carbonyl)amino)-3-hydroxycyclohexyl)oxy)-5-azido-3-(benzyloxy)tetrahydro- 2H-pyran-2-yl)methyl)(methyl)carbamate (654 mg, 0.566 mmol), DIPEA (0.242 mL, 1.42 mmol) and (2S)-4- (benzyloxycarbonylamino)-2-hydroxy-butanoic acid (215 mg, 0.849 mmol) in dry DMF (10.9 mL) at 22 °C. The mixture was stirred at 22 °C for 25 min and was diluted with sat. aq. NH₄Cl (20.0 mL), EtOAc (30.0 mL) and brine (100 mL). The aqueous phase was extracted with EtOAc (3 x 50.0 mL), and the combined organic extracts were washed with brine (50.0 mL), dried (MgSO₄), filtered, and concentrated. The residue was purified by silica gel chromatography (24 g high-capacity cartridge) using a gradient of EtOAc in hexanes (50-100%) and a gradient of EtOAc in MeOH (0-20%) to provide the title compound as a solid (460 mg, 49%). m/z (ES+) [M-CD ring+H]⁺ = 1047.7; HPLC (A05) t_R = 1.81 min. AttyDktNo. R1119431100WO Step B. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,5S,6R)-3-amino- 5-hydroxy-6-((methylamino)methyl)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 8) [00406] A mixture of benzyl (((2R,3S,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-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)oxy)-5-(benzyloxy)-6-(((benzyloxy)carbonyl)amino)-4-((S)-4- (((benzyloxy)carbonyl)amino)-2-hydroxybutanamido)-3-hydroxycyclohexyl)oxy)-3-(benzyloxy)tetrahydro- 2H-pyran-2-yl)methyl)(methyl)carbamate (460 mg, 0.331 mmol) in MeOH (14.3 mL) was added to a suspension of Pd(OH)₂ (0.697 g, 0.993 mmol, 20% wet) in AcOH (11.7 mL) and water (4.62 mL). The mixture was evacuated with hydrogen three times and stirred at 22 °C under continuous bubbling of hydrogen balloon pressure for 4 h. The mixture was evacuated with nitrogen, bubbled with nitrogen for 5 min. The suspension was filtered on Celite, washed with water (100 mL), MeOH (100 mL), water (2 x 50.0 mL). The volatiles were evaporated under reduced pressure (rotavapor bath temperature < 35 °C). The material was purified by preparative HPLC using 5% of MeCN in water ^10 mM Ammonium bicarbonate pH 10 ^, and lyophilized to give a solid. The solid was dissolved in water (10.0 mL), and (NH₄)₂SO₄ (131 mg, 0.993 mmol) was added. The mixture was stirred at 22 °C for 15 min, then filtered on a medium frit and added dropwise to MeOH (400 mL) under vigorous stirring. The suspension was centrifuged (rotation speed: 1765 rpm) for 1 h. MeOH supernatant was carefully removed, and the solid was dissolved in distilled water (25.0 mL). The solution was evaporated under reduced pressure to remove residual MeOH, and lyophilized to provide the title compound (Compound 8) as a solid (280 mg, 83%, >99% purity by qNMR).¹H NMR (400 MHz, D₂O) δ 5.94 (d, J = 3.7 Hz, 1H), 5.41 (d, J = 1.8 Hz, 1H), 5.30 (d, J = 1.8 Hz, 1H), 4.55 (dd, J = 7.3, 4.7 Hz, 1H), 4.45 (dd, J = 4.7, 1.8 Hz, 1H), 4.39 (dd, J = 8.1, 4.1 Hz, 1H), 4.35 (ddd, J = 7.0, 3.9, 1.6 Hz, 1H), 4.30 – 4.18 (m, 3H), 4.01 – 3.83 (m, 6H), 3.78 – 3.65 (m, 4H), 3.60 (dt, J = 3.0, 1.3 Hz, 1H), 3.51 – 3.33 (m, 4H), 3.28 – 3.14 (m, 3H), 2.79 (s, 3H), 2.32 – 2.14 (m, 2H), 2.13 – 2.01 (m, 2H). m/z (ES+) [M+H]⁺ = 730.6; HPLC (A05) t_R = 0.123 min. Example 11: Minimum Inhibitory Concentration Assays [00407] Minimum inhibitory concentrations for representative example compounds herein were determined by a broth microdilution method in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines. In brief, organism suspensions were adjusted to a 0.5 McFarland standard to yield a final inoculum between 3 ^10⁵ and 7 ^10⁵ colony-forming units (CFU)/mL. The inoculum was prepared by suspension of a colony from an agar plate that was prepared the previous day. Compound dilutions and inoculum were prepared in sterile, cation adjusted Mueller-Hinton Broth (Beckton Dickinson). [00408] Bacteria were suspended in sterile saline and added to each assay plate to obtain a final concentration of 5 ^10⁵ CFU/mL. An inoculum volume of 100 µL was added to wells containing 100 µL of broth with 2- fold serial dilutions of the test compound. All inoculated microdilution trays were incubated in ambient air at AttyDktNo. R1119431100WO 35°C for 18-24 hours. Following incubation, the lowest concentration of the compound that prevented visible growth (OD 600 nm < 0.05) is recorded was the MIC. The MIC was determined to be the lowest concentration of the test compound that resulted in no visible bacterial growth as compared to an untreated control. Performance of the assay was monitored by use of laboratory quality-control strains and a standard compound with a defined MIC spectrum (neomycin), in accordance with CLSI guidelines. [00409] Compounds of the disclosure (Compounds 1 - 8) were evaluated against multiple strains of A. baumannii, E. coli, and P. aeruginosa. Results are provided in Table 1 and Table 2 below, which demonstrate that all compounds are active against the majority of the strains evaluated, with MIC values generally equal to or less than 16 ^g/ml. Importantly, all compounds retain potent activity against an E. coli strain (NECO0245) that expresses the ribosomal methyl transferase ArmA, which is known to confer resistance to all clinically relevant parenteral aminoglycoside antibiotics.

_{. 8 + + + + + + + + + + + + + +} ^D _. N _{+ . 7 + + + + + + + + + + + + + +} ^D _. N _{+ . 6 + + + + + + + + + + + + + +} ^D _. N ₊ yt_i ^v _{i .} t_{c 5 + + + + + + + + + + + + + +} ^D _{. +} ^B A N d ⁿ _i n ^c a _{. y # 4 + + + + + + + + + + + + + +} ^D _{. +} m d N ^{o n} _{e u} ^{n o} _{p .} ^{= m} _{3 + + + + + + + + + + + + + +} ^D _. ^O o N _{+ E} C ^N _; ⁾ 2 _{L + + + + + + + + + + + + + + + +} ^m _/ ^g _{µ 6 1 + + + + + + + + + + + + + + + +} ^{1 > s} i )- ^O _{( E + + + + + + - + + - - + + + + + e} N ^v _i ^t _c ^a iⁱ _{i i a a a a a n} ⁱ n ⁱ _n ⁱ _n ^s _o ^s _o ^{s s s} _r n s _a ⁿ _a ⁿ _a ⁿ _i e_i m ^{g n} _{i o} g ⁿ _{i o} g ⁿ _{i o} ⁿ _i ^o _f d ^{c u} m^u m^u _{i e a} ^l _i ^l _{i u u u} ^g _u ^g _{u i i i i i n} p ^b _a ^b _a ^b _o ^c _o ^{l c} _o ^{r c} _e ^{r a} _e ^{r a} _e ^{r a} _e ^{r a} _e ^{l a} _o ^{l l l l a c} _o ^c _o ^c _o ^c _o ^{c ,} L _{- S} ^. _A ^. _A ^. _A ^. _E ^. _E ^. _E ^. _P ^. _P ^. _P ^. _P ^. _P ^. _E ^. _E ^. _E ^. _E ^. _E ^{m 8} / ₀ g ¹ µ _- b_II- _{6 l )’} ^{1 e} _n ^{3 a} ₍ ^≤ s _P ^h _p ⁱ _{) y} ^{A r} _, ⁺ _a ^b _I ^d ( e ^{) y m}i _{n r} ^{i P} _{a C I}- ^{rt Q} _)’ ^{s 3} _s ^e _R t t _{i e} ^v _i ^{t – x} _{s , e} ^bI _I ^bI _{( r I} ^h _p ^p _x ^t ( _c e O ^{a a} _t ^a _u ^l _f ^vi_t ^-) _’ ^-) _’ ^A _{, r} ^{e Kr} _{o b a} ^e _{v d e} ^{p t i} _{y y} ^{g t} _n ^e _f ^{a 3} r ^e _g ^e ₍ ^{3 h} ₍ ^h O_v K ^{o b} _I ^{t I} _{- c} ^I- ^I- ^II t_{i )} ^e _{v )} ^’ ₎ ^’ -^{) s} _{o o o} ^{a l n} _{e p} ^K _{J n C} Q _{p p x} Y X^’ _{y 6} ⁽ ₂ ⁽ ₃ ⁽ A ^p T ^e _{d E} ^C _l AA^ul _{3 f} ^{x (} _h ^t _{p C C C} ^m _{r o} ^{i h} _P M_{T T} ^o _{T T e} AAA W^a _Δ A ^{t f p m} _r ^o _{f b} W W _Δ W W _E M _{a e} A A A A^o _{d r} ^{l c} _{o i e} ^{6 7 h M d .} _{o 0 1} ^{C 0} ₀ ^{0 0} ₂ ^{1 0} ₀ ^{2 0} ₀ ^{3 0} ₀ ⁰ ₁ ⁰ _{2 6 0 9} ³ 0 ⁰ ₀ ⁰ ₀ ¹ ₀ ⁵ ₄ ^{4 2} ₄ ^{0 2 5} _s 2 ₅ ³ ₀ ³ ₄ ^{2 e} _r n ₀ A₀ A_{0 0 0 0} ^{0 0 0 0} ₃ ⁰ _{0 0 0 0 0} ^h t _e ^l _{i b} ^a _{r B} A_B ^A A_BO A^CO E ^CO E ^C _{E E E E E}O E ^A _P ^A _P ^A _P ^A _P ^A _P ^CO E ^CO CO CO C ^C _I ^{a t} _S N N N N N _{E E E E} M _T N N N N N N N N N N N _{* e} ^d _o C ⁿ _i ^ar_t ^{S k} _n ^a B n_{i e} c ^t _{a y} ^l m^o _{- - + - + - - - + - - - - - - - + - + - + + + + + + + + - + + + o} ^s I e ⁾ _* N _R A ^y (t_i ^{e v} _{c i} ^{t n} _{c a} ^t A _{2 + + + + + + + + + + + + - + - + + + + + + + + + + + + + + + + + si d} ^s _e ⁿ _a ^R _# ^ci_{t d} ^o _i ^{n b} _{u i} ^{t o} _{n p 1 + + + + + + + + + - + + + + + + + + + + +} A^m _{+ + + + + + + + + + + o C} D C ^C _* ^{* * a} _* ^* _* ^* _* ^* _* ^{* )} * ^* _* ^{* * * * * *} _{L k} ⁸ ₁ ^{9 7 8 9 3} _{* * * * * *} ^m _/ 1 ₁ ¹ ₂ ¹ ₂ ¹ ₃ ^{8 3 7 9 1 4 1} _{4 4 5 5 5 6 5} ^g A _{0 0 0 0 0} ¹ ₀ ¹ ₀ ¹ ₀ ¹ ₀ ¹ ₀ ¹ ₀ ¹ _{0 µ 6} ¹ iⁱ _{i n} ⁱ _i ⁱ _i ⁱ _i ⁱ _i ⁱ _i ⁱ _i ⁱ _i ⁱ _i ⁱ _a ^s _a ^s _a ^s _a ^s _a ^s _a ^s _a ^s _a ^s _a ^s _a ^s _e ^a _e ^> i ^a _{e i} ^a _{e i} ^a _i ^{s s} _i ^{) n} _n ⁿ _n ⁿ _n ⁿ _n ⁿ _n ⁿ _n ⁿ _n ⁿ _n ⁿ _n ⁿ _o ⁿ _o ⁿ _o ⁿ _o ⁿ _{o o o o o o a} ⁿ _o ⁿ _o ⁿ _o ⁿ _{o i} ^{i si} _{l e} ^{n e} _{-( s a a a a a a a a a a i} ^e _i m ^g _i ^g _i ^g _i ^{n g} _i ^{n g} _i ^{n g} _i ^{n g} _i ^{n g} _i ^{n g} _i ^{g c} _i ^mmmm _d ^{n i} _{b e} ^g _a ^c _e ^{v c u} m _{e a} ^u m a ^u m a ^u m a ^u m a ^u m a ^u m a ^u m a ^u m a ^u _{u u u u u u u u u u a} ^r _e ^r _e ^r _e ^r _e ^r _e ^r _e ^r _e ^r _e ^r _e ^r _{e i} ^l _i ^r o ^l _{o e} ^t _{i u} ^{p b} _n ^l _o ^e _{u n} ^e _u ^e _u ^e _u ^{a p} _{n n n} ^e _{r ri o} ^r _{e a} ^o _{i l} ^t _c ^a _S ^{. b} A ^{. b} A ^{. b} A ^{. b} A ^{. b} A ^{. b} A ^{. b} A ^{. b} A ^{. b} A ^{. a} A ^{. a} P ^{. a} P ^{. a a a a a a a c c e c} P ^. _P ^. _P ^. _P ^. _P ^. _P ^. _P ^. _P ^. _E ^. _E ^. _S ^. _E ^{. p} K^{. p} K^{. p} K^{. f} K_. ^{m a} C^. _P ^{. c} _{n K} ^. _E ⁱ _r ^o f d -⁹ )_l ^e )_l )_l )_l )_l )_l )_l )_l )_l )_l )_l ) _n ^a _{0 n} ^{e a} _n ^{e a} _n ^{e a} _n ^{e a} _n ^{e a} _n ^{e a} _n ^{e a} _n ^{e a} _n ^e _l a _n ^{e a} _n ^{e , 1 a} _{n L - p p p p p p p p p p p} ^a _p ^m E _{E E E E E E E E E /} ^{g R R R R R R R R R R} _{E E} R _{µ C C C C C C C C C C} ^R _{C C 6 C C C C C C C C C C C} ¹ DDDDDDDDD_C D D ^{≤ C C C C C C C C C C}D C _s ⁱ e^t _{a e} l ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e} ^t _{a e (} t_{a e (} t_{a e (} t_{a e (} t_{a e (} t_{a e (} t_{a e (} t _{e (} t _{e (} t _{e (} t _{e (} ^{C t} _{e (} t _{e )} t ⁺ ( o ^{l s} _{i o} ^{l s} _o ^{l s} _o ^{l s} _o ^{l s} _o ^{l s} _o ^{l s} _o ^{l s} _o ^{l s} _o ^{l s} _o ^{l l l l l l l l l l l l l l} _a ^l _a ^l _a ^l _a ^l _a ^l _a ^l _a ^{l y s} _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^s _o ^{s t} _i l _{i a} ^l _i ^l _i ^l _i ^l _i ^l _i ^l _{i i i} ^{i i i i i i i i i i il il il il il il il il il} _i ^{il il v} _i ^t _e ^{c p} _{i a y} ^{n c} _{i a} ^c _{i a} ^c _{i a} ^c _{i a} ^c _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^{l c} _{i a} ^c _{i a} ^{l c} _{i a} ^c _{i a} ^c _{i a} ^c _{i a} ^c _{i a} ^c _{i a} ^c _a ^c _a ^c _a ^c _a ^c _a ^c _c ^{a t} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _i ^{l n} _{i i} l ⁿ _{i i} l ⁿ _{i i} l ⁿ _{i i} l ⁿ _{i i} ^{l n} _{i i} l ⁿ _i ^e _v ⁱ _o ⁿ _{C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C} ^l Ct_{i e R} ^h D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R ^s _o D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D_R D ^p _P M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M_r ^o _{f d} ^{l 7} ₂ ^{3 4} ₃ ⁴ ₃ ⁶ ₃ ⁷ ₃ ⁸ ₃ ⁹ ₃ ⁸ ₄ ⁹ ₄ ⁰ _{o 5 8} ⁸ ₃ ⁹ ₉ ⁹ ₂ ⁰ ₃ ⁰ ₈ ⁰ ₀ ¹ ₃ ¹ ₆ ¹ ₃ ^{2 5} ₂ ⁶ _{2 1} ⁷ ₂ ⁶ ₄ ⁰ ₅ ³ ₅ ⁵ _{5 1} ⁰ _{8 2} ⁴ ₉ ^{8 h} _{s n} ⁱ ₁ ⁴ ₁ ⁴ ₁ ⁴ ₁ ⁴ ₁ ⁴ ₁ ⁴ ₁ ⁴ ₁ ⁴ ₁ ⁴ _{1 2 2 2 3 3 3 3 3 3 3} ² ₁ ² ₁ ⁰ ₀ ² ₁ ² ₁ ² ₁ ² ₁ ² _{1 0} ⁴ _{0 0 0} ^e _{r a e} ^AAAAAAAAAA1 _E ¹ _E ¹ _E ¹ _E ¹ _E ¹ _E ¹ _E ¹ _E ¹ _E ¹ _EOO¹ _EO^NNNN1 _P ¹ _I ¹ _E ¹ _L ^h _t ^r _t ^{d S} _{o B} A_B A_B A_B A_B A_B A_B A_B A_B A_B A^A C N N N N N N N N N N_P ^A N_P ^A N_P ^A N_P ^A N_P ^{AAAAAC C S C} _{P P P P} ^{S MAC C} _I N_P N_P N_P N_P N_P N_E A_E A_S A_EKKKK A A A A A_C A_P A_E A_E AM * AttyDktNo. R1119431100WO Example 12. Microbiology surveillance studies [00410] A large-scale microbiology surveillance study was used to determine the antibacterial activity of compounds against contemporary Gram-negative clinical isolates from Europe and North America. The surveillance panel was composed of 170 Enterobacterales, 50 P. aeruginosa, and 50 A. baumannii isolates collected in 2020 from locations in the Europe and North America (1:1 distribution). The MIC was determined to be the lowest concentration of the test compound that resulted in no visible bacterial growth as compared to an untreated control. Performance of the assay was monitored by use of laboratory quality- control strains in accordance with CLSI guidelines. [00411] The MIC₅₀ is defined as the concentration of drug at which the growth of 50% of a given set of isolates is inhibited. The MIC₉₀ is defined as the concentration of drug at which the growth of 90% of a given set of isolates is inhibited. [00412] Certain compounds of the disclosure (Compounds 1 and 2) were evaluated against the large-scale microbiology surveillance panel of Enterobacterales, P. aeruginosa, and A. baumannii. Results are provided in Table 3 below, which demonstrates that Compounds 1 and 2 are active against the majority of the strains evaluated with MIC₅₀ and MIC₉₀ values equal to or less than 4 µg/mL for both compounds. Table 3. Microbiology surveillance data Compound # and Activity* Organism Group n 1 2 MIC₅₀ MIC₉₀ MIC₅₀ MIC₉₀ Enterobacterales 170 A A A A P. aeruginosa 50 A A A A A. baumannii - calcoaceticus 50 A A A A *A = MIC_50/90 ≤ 4; B = MIC_50/90 > 4 Example 13. In vivo kidney accumulation studies [00413] The measured drug concentrations in rat kidneys, normalized to the plasma pharmacokinetics area under the curve, indicates the extent of accumulation in the kidneys. Without wishing to be bound by theory, it is believed that aminoglycosides with lower nephrotoxic potential are generally less likely to demonstrate high kidney accumulation, and lower accumulation is therefore believed to correlate with a better safety profile with respect to nephrotoxicity. Example 14. In vivo repeat-dose nephrotoxicity studies [00414] Compounds are evaluated for nephrotoxic potential via a repeat-dose study in rats. Individual drugs are administered to rats via subcutaneous or intravenous injection once daily for multiple sequential days. Blood samples are analyzed for biomarkers of nephrotoxicity such as serum creatinine, urea, and BUN. At terminal time-points, the rat kidneys are resected, fixed, and tissues analyzed by histopathology and AttyDktNo. R1119431100WO immunochemistry to determine No Observed Adverse Effect Level (NOAEL) value for each compound. Without wishing to be bound by theory, it is believed that seemingly small structural modifications on the aminoglycoside ABCD ring system (e.g., removing or altering substituents) may result in significant differences in nephrotoxicity independent of any potential changes in antibacterial activity.

Claims

AttyDktNo. R1119431100WO CLAIMS What is claimed is: 1. A compound having a structure according to Formula I: ,

or a pharmaceutically or R¹ and R² are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R³ and R⁴ are each independently selected from the group consisting of hydrogen and C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, -OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S, =N(R⁶), and -CN; R⁵ is hydrogen or is C_1-4 alkyl, wherein said C_1-4 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, - OR⁶, -SR⁶, -C(O)N(R⁶)₂, -N(R⁶)₂, -S(O)R⁶, -S(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -OC(O)R⁶, -NO₂, =O, =S,

C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl, wherein each of said C_1-10 alkyl, C_2-10 alkenyl, and C_2-10 alkynyl is optionally substituted with one or more substituents independently selected for each occurrence from halogen, -OH, -CN, -NO₂, and -NH₂; R⁷ is selected from the group consisting of hydrogen, -OR⁶, and -N(R⁶)₂; R⁸ is selected from the group consisting of hydrogen and -OR⁶; R^a is a tetrahydrofuranyl ring or a hexahydropyranyl ring, connected via a glycosidic bond; AttyDktNo. R1119431100WO R^b is a second tetrahydrofuranyl or hexahydropyranyl ring connected to R^a through a glycosidic bond; wherein each of R^a and R^b is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OR⁶, -N(R⁶)_2, and C_1-6 alkyl, wherein said C_1-6 alkyl is optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and R^c is hydrogen, C_1-6 alkyl, a protecting group, or has a structure according to Formula II: , wherein:

Q is NH, O, or S; n is an integer from 0 to 4; R⁹ is hydrogen or C_1-3 alkyl; R¹⁰ and R¹¹ are each selected independently for each occurrence from the group consisting of hydrogen, alkyl, halogen, and –OH; and R¹² is H, C_1-3 alkyl, or –C(=NH)NR^12aR^12b, wherein R^12a and R^12b are each independently selected from the group consisting of hydrogen and C_1-3 alkyl; or R⁹ and R¹², together with the atoms to which they are attached, form a heterocycloalkyl ring system comprising at least one N. 2. The compound of claim 1, wherein R¹ and R² are both hydrogen. 3. The compound of claim 1, wherein R¹ and R² are both methyl. 4. The compound of claim 1, wherein one of R¹ and R² is hydrogen, and the other of R¹ and R² is methyl. 5. The compound of any one of claims 1 to 4, wherein R³ is hydrogen and R⁴ is methyl. 6. The compound of any one of claims 1 to 4, wherein R¹, R² and R³ are hydrogen, and R⁴ is methyl. AttyDktNo. R1119431100WO 7. The compound of any one of claims 1 to 4, wherein R¹ and R³ are both hydrogen and R² and R⁴ are both methyl. 8. The compound of any one of claims 1 to 7, wherein R⁵ is hydrogen. 9. The compound of any one of claims 1 to 8, wherein R^c is selected from the group consisting of: .

10. The compound of any one to .

11. The compound of any one of claims 1-10, having a structure according to Formula Ia: , wherein:

R¹³ and R¹⁴ are each independently selected from –OR⁶ and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, C_1-6 alkyl, -OR⁶, -N(R⁶)₂, -NO₂, and –CN; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of –OR⁶, N(R⁶)₂, and C_1-6 alkyl, said C_1-6 alkyl optionally substituted with one or more AttyDktNo. R1119431100WO substituents independently selected from the group consisting of halogen, -OR⁶, -N(R⁶)₂, -NO₂, and –CN. 12. The compound of claim 11, wherein: R¹³ is H or OH; and R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl. 13. The compound of claim 12, wherein R' and R'' are each hydrogen. 14. The compound of claim 12, wherein R' is hydrogen and R'' is methyl. 15. The compound of claim 12, wherein R' and R'' are each methyl. 16. The compound of claim 12, wherein R¹³ is -OH and R¹⁴ is -CH₂OH. 17. The compound of claim 11, having a structure according to Formula Ib: , wherein: 1

R is hydrogen or R⁷ is NH₂ or OH; R¹⁴ is –C(R'R'')OH, where R' and R'' are each independently selected from hydrogen and methyl; and R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are each independently selected for each occurrence from the group consisting of -OH, -NH₂, and C_1-3 alkyl, said C_1-3 alkyl optionally substituted with one or more substituents independently selected for each occurrence from the group consisting of -OH, -NH₂, -NO₂, and -CN. AttyDktNo. R1119431100WO 18. The compound of claim 17, wherein R⁷ is NH_2. 19. The compound of claim 17, having a structure according to Formula Ic: .

20. The compound of a structure .

AttyDktNo. R1119431100WO 21. The compound of claim 20, wherein R^c is: OH OH NH₂ NH_{2 .}

22. The compound a structure to OH H N NH ^c R¹ ₂ R , wherein R¹ is hydrogen or

23. The compound of claim 22, having a structure selected from: .

AttyDktNo. R1119431100WO 24. The compound of claim 23, wherein R^c is: .

25. A any one of claims 1-24, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, and at least one pharmaceutically acceptable excipient. 26. A method for treating a bacterial infection in a subject in need thereof, the method comprising administering to the subject a compound of any one of claims 1-24, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or the pharmaceutical composition of claim 25. 27. The method of claim 26, wherein the bacterial infection is with Gram-positive, Gram-negative, an aerobic, or facultative anaerobic bacteria. 28. The method of claim 26, wherein the bacterial infection is with a member of the order of Enterobacterales. 29. The method of claim 26, wherein the bacterial infection is with an Escherichia spp., a Klebsiella spp., a Proteus spp., a Citrobacter spp., a Morganella spp., a Providencia spp., a Yersinia spp., an Enterobacter spp., a Salmonella spp., or a Serratia spp. 30. The method of claim 26, wherein the bacterial infection is with a Moraxella spp., a Pseudomonas spp., an Acinetobacter spp., a Mycobacterium spp., a Staphylococcus spp., a Bacillus spp., a Francisella spp., or a Burkholderia spp.