Broad spectrum aminoglycosides and uses thereof

Abstract

# imgabs0 # provides herein aminoglycoside compounds, such as compounds of Formula (I) and Formula (III), and pharmaceutically acceptable salts thereof, for use as therapeutic and / or prophylactic agents. The compounds are useful in the treatment of bacterial infections in a subject, such as gram-negative bacterial infections. Also provided herein are methods of making such compounds.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 316,731, filed on March 4, 2022, which is incorporated herein by reference in its entirety.

[0003] Statement Regarding Federal Funding

[0004] Research or development

[0005] 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

[0006] The present disclosure relates generally to aminoglycoside compounds having antimicrobial activity and to their use in treating medical conditions associated with pathogenic microorganisms. Background Art

[0007] The rapid spread of antibiotic resistance has prompted a continued search for new agents that can have antibacterial activity. Aminoglycosides (AGs) are highly potent, broad-spectrum antibiotics with a variety of desirable properties for treating life-threatening infections. AGs are used to treat a variety of diseases, including but not limited to: urinary tract infection, bloodstream infection, pneumonia and neonatal sepsis. Examples of AGs include: tobramycin, gentamicin and amikacin. AG antibiotics exert their antibacterial effects by binding to specific targeting sites in bacterial ribosomes. For the structurally related antibiotics neamine, ribostamycin, neomycin B and paromomycin, the binding site has been located at the A site of the prokaryotic 16S ribosomal decoding region RNA (see Moazed, D.; Noller, HF, Nature, 1987, 327, 389). Binding of AG to this RNA target interferes with the fidelity of mRNA translation and leads to miscoding and truncation, ultimately causing bacterial cell death (see Alper et al., J. Am. Chem. Soc., 1998, 120, 1965).

[0008] Over time, resistance to AG has emerged clinically and has weakened our antimicrobial armamentarium. AG resistance (AG-R) can be divided into two main types mechanistically: (1) AG-modifying enzymes (AMEs), which modify and inactivate drugs and (2) enzymes called ribosomal methyltransferases (RMTs), which modify the drug target binding pocket. Currently, the most common form of AG-R in clinical isolates is conferred by AMEs, which are diverse and widely distributed. Although less common than AMEs in some species, RMTs are a serious threat because they alter the 16S rRNA AG binding site and prevent AG target binding, conferring high-level resistance to all clinically available parenteral AGs. Another problem is that AME and RMT genes are often located on plasmids or transposons together with genes encoding resistance to other classes of antimicrobial agents, resulting in multidrug-resistant isolates.

[0009] In addition, AG can be toxic to the kidneys (nephrotoxicity) and to the cochlea (ototoxicity). These are cumulative processes, and the likelihood of observing toxicity increases with the duration of treatment. The nephrotoxic potential of AG limits the dose and length of treatment, making it challenging to achieve the systemic exposure required for efficacy against certain infections. Many times, but not always, the structure-activity relationship that leads to increased antibacterial efficacy is associated with the structure-activity relationship that leads to toxicity to human kidney cells. It is speculated that the source of this toxicity is the result of a combination of different factors and mechanisms, such as interaction with phospholipids, inhibition of phospholipases, and the formation of free radicals.

[0010] For the reasons mentioned above, while progress has been made in this field, new chemical entities with antimicrobial activity against highly resistant organisms and with an acceptable safety profile are needed. Summary of the invention

[0011] The present disclosure relates to aminoglycoside compounds having antibacterial activity, including stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof, and to their use, for example, in treating conditions associated with pathogenic microorganisms (also referred to herein as "bacterial infections").

[0012] In one aspect, provided are compounds having a structure according to Formula (I):

[0013]

[0014] or a pharmaceutically acceptable salt or solvate thereof,

[0015] in:

[0016] R a is hydrogen (H) or methyl; and

[0017] R b C 1-6 Alkyl, or having a structure according to formula (II):

[0018]

[0019] in:

[0020] Q is NH, O or S;

[0021] n is an integer from 0 to 4;

[0022] R 1 is hydrogen or C 1-3 alkyl;

[0023] R 2 and R 3 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, halogen, and -OH; and

[0024] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b Each independently selected from H and C 1-3 Alkyl; or

[0025] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

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

[0027] In some embodiments, R a It is methyl (CH3).

[0028] In some embodiments, R b Selected from:

[0029]

[0030] In some embodiments, R b Selected from:

[0031]

[0032] In some embodiments, the compound of formula (I) has a structure selected from the following:

[0033]

[0034] In some embodiments, R b It is ethyl (C2H5).

[0035] In some embodiments, the compound of formula (I) has the following structure:

[0036] In another aspect, provided are compounds having a structure according to formula (III):

[0037] in:

[0038] R a is H or methyl; and

[0039] R b H, C 1-6 Alkyl, or having a structure according to formula (II):

[0040] in:

[0041] Q is NH, O or S;

[0042] n is an integer from 0 to 4;

[0043] R 1 is hydrogen or C 1-3 alkyl;

[0044] R 2 and R 3 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, halogen, and -OH; and

[0045] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b are each independently selected from hydrogen and C 1-3 Alkyl; or

[0046] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0047] In some embodiments, R a For H.

[0048] In some embodiments, R a It is methyl.

[0049] In some embodiments, the compound of formula (III) has a structure selected from the following:

[0050]

[0051] In some embodiments, R b It is ethyl (C2H5).

[0052] In some embodiments, the compound of formula (III) has a structure selected from the following:

[0053]

[0054] In some embodiments, R b Selected from:

[0055]

[0056] In some embodiments, R b Selected from:

[0057]

[0058] In some embodiments, the compound of formula (III) has a structure selected from the following:

[0059]

[0060] In another aspect, a pharmaceutical composition is provided, comprising a compound of formula (I), a compound of formula (III), or a pharmaceutically acceptable salt or solvate of any one thereof, and at least one pharmaceutically acceptable excipient.

[0061] In a further aspect, a method for treating a bacterial infection in a subject in need thereof is provided, the method comprising administering to the subject a compound of any one of formula (I) or (III), or a pharmaceutically acceptable salt or solvate of any one of them, or a pharmaceutical composition comprising the compound. In some embodiments, the method comprises administering to the subject a compound of formula (I) or formula (III), or a pharmaceutically acceptable salt or solvate of any one of them, or a pharmaceutical composition comprising the compound.

[0062] In some embodiments, the bacterial infection is caused by gram-positive bacteria, gram-negative bacteria, aerobic bacteria, 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.

[0063] In some embodiments, the bacterial infection is caused by Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, acter, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Burkholderia, or Enterococcus.

[0064] In some embodiments, the bacterial infection is an infection caused by Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, Proteus mirabilis, Bacillus anthracis, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, mallei), Burkholderia pseudomallei, or Yersinia pestis.

[0065] In some embodiments, the bacterial infection is a gram-negative bacterial infection. In some embodiments, the gram-negative bacterial infection is an infection caused by Pseudomonas aeruginosa, Stenotrophomonas maltophila, Burkholderia cepacia, or Alcaligenes xylosoxidans.

[0066] In some embodiments, the bacterial infection is caused by multidrug resistant (MDR) bacteria.

[0067] In some embodiments, the bacterial infection is an infection caused by an extended spectrum beta-lactamase (ESBL)-producing or carbapenem-resistant Enterobacteriaceae.

[0068] In some embodiments, the bacterial infection is caused by Staphylococcus aureus. In some embodiments, the Staphylococcus aureus is methicillin-resistant Staphylococcus aureus (MRSA).

[0069] The present disclosure includes, but is not limited to, the following embodiments.

[0070] Embodiment 1. Compounds having a structure according to formula (I):

[0071]

[0072] or a pharmaceutically acceptable salt or solvate thereof,

[0073] in:

[0074] R a is hydrogen (H) or methyl; and

[0075] R b C 1-6 Alkyl, or having a structure according to formula (II):

[0076]

[0077] in:

[0078] Q is NH, O or S;

[0079] n is an integer from 0 to 4;

[0080] R 1 is hydrogen or C 1-3 alkyl;

[0081] R 2 and R 3 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, halogen, and -OH; and

[0082] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b Each independently selected from H and C 1-3 Alkyl; or

[0083] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0084] Embodiment 2: A compound as described in embodiment 1, wherein R a For H.

[0085] Embodiment 3: A compound as described in embodiment 1, wherein R a It is methyl.

[0086] Embodiment 4: A compound as described in any one of embodiments 1-3, wherein R b Selected from:

[0087]

[0088] Embodiment 5: A compound as described in any one of embodiments 1-3, wherein R b Selected from:

[0089]

[0090] Embodiment 6: A compound as described in embodiment 1, wherein the compound of formula (I) has a structure selected from the following:

[0091]

[0092] Embodiment 7: A compound as described in any one of embodiments 1-3, wherein R b It is ethyl.

[0093] Embodiment 8: The compound as described in Embodiment 7, wherein the compound of formula (I) has the following structure:

[0094]

[0095] Embodiment 9: Compounds having a structure according to formula (III):

[0096]

[0097] in:

[0098] R a is H or methyl; and

[0099] R b H, C 1-6 Alkyl, or having a structure according to formula (II):

[0100]

[0101] in:

[0102] Q is NH, O or S;

[0103] n is an integer from 0 to 4;

[0104] R 1 is hydrogen or C 1-3 alkyl;

[0105] R 2 and R 3 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, halogen, and -OH; and

[0106] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b are each independently selected from hydrogen and C 1-3 Alkyl; or

[0107] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0108] Embodiment 10: A compound as described in Embodiment 9, wherein R a For H.

[0109] Embodiment 11: A compound as described in Embodiment 9, wherein R a It is methyl.

[0110] Embodiment 12: A compound as described in any one of embodiments 9-11, wherein the compound of formula (III) has a structure selected from the following:

[0111]

[0112] Embodiment 13: A compound as described in any one of embodiments 9-12, wherein R b It is ethyl.

[0113] Embodiment 14: A compound as described in Embodiment 13, wherein the compound of formula (III) has a structure selected from the following:

[0114]

[0115] Embodiment 15: A compound as described in any one of embodiments 9-12, wherein R b Selected from:

[0116]

[0117] Embodiment 16: A compound as described in any one of embodiments 9-12, wherein R b Selected from:

[0118]

[0119] Embodiment 17: A compound as described in Embodiment 16, wherein the compound of formula (III) has a structure selected from the following:

[0120]

[0121]

[0122] Embodiment 18: A pharmaceutical composition comprising the compound according to any one of Embodiments 1 to 17 or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, and at least one pharmaceutically acceptable excipient.

[0123] Embodiment 19: A method for treating a bacterial infection in a subject in need thereof, the method comprising administering to the subject a compound as described in any one of Embodiments 1-17 or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or administering a pharmaceutical composition as described in Embodiment 18.

[0124] Embodiment 20: The method of embodiment 19, wherein the bacterial infection is caused by Gram-positive bacteria, Gram-negative bacteria, aerobic bacteria, or facultative anaerobic bacteria.

[0125] Embodiment 21: A method as described in embodiment 19, wherein the bacterial infection is an infection caused by: Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Burkholderia, or Enterococcus species.

[0126] Embodiment 22: A method as described in embodiment 20 or 21, wherein the bacterial infection is an infection caused by the following species: Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter krusei, Proteus mirabilis, Bacillus anthracis, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei, or Yersinia pestis.

[0127] Embodiment 23: The method of any one of Embodiments 19 to 22, wherein the bacterial infection is a Gram-negative bacterial infection.

[0128] Embodiment 24: The method of embodiment 23, wherein the Gram-negative bacterial infection is an infection caused by Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Burkholderia cepacia, or Alcaligenes xylosoxidans.

[0129] Embodiment 25: The method of any one of embodiments 19-24, wherein the bacterial infection is caused by a multidrug resistant (MDR) bacteria.

[0130] Embodiment 26: The method of any one of embodiments 19-22, wherein the bacterial infection is an infection caused by an extended-spectrum beta-lactamase (ESBL)-producing bacterium or a carbapenem-resistant Enterobacteriaceae.

[0131] Embodiment 27: The method of any one of embodiments 19-25, wherein the bacterial infection is caused by Staphylococcus aureus.

[0132] Embodiment 28: The method of embodiment 27, wherein the Staphylococcus aureus is methicillin-resistant Staphylococcus aureus (MRSA).

[0133] By reading the specific embodiments below, these and other features, aspects and advantages of the present disclosure will become clear. The present disclosure includes any combination of two, three, four or more features or elements set forth in the present disclosure, regardless of whether these features or elements are explicitly combined or otherwise referenced in the specific exemplary embodiments described herein. The present disclosure is intended to be understood as a whole so that any separable features or elements of the present disclosure, in any aspect and exemplary embodiment thereof, should be considered combinable, unless the context of the present disclosure clearly provides otherwise.

[0134] Therefore, it is understood that the present disclosure is provided only to summarize some exemplary embodiments to provide a basic understanding of some aspects of the present disclosure. Therefore, it should be understood that the above exemplary embodiments are merely examples and should not be interpreted as narrowing the scope or spirit of the present disclosure in any way. Other exemplary embodiments, aspects and advantages will be clear from the following specific embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0135] Having generally described various aspects of the present disclosure above, the present disclosure will now be described with reference to the accompanying drawings, which are not necessarily drawn to scale. The accompanying drawings are merely exemplary and should not be construed as limiting the present disclosure.

[0136] Figure 1 A non-limiting scheme for preparing a compound of the present disclosure having a structure of Ring A1 according to one embodiment is shown;

[0137] Figure 2A non-limiting scheme for preparing a compound of the present disclosure having a structure of Ring A2 according to one embodiment is shown;

[0138] Figure 3 A non-limiting scheme for preparing a compound of the present disclosure having a structure of Ring A3 according to one embodiment is shown;

[0139] Figure 4 A non-limiting flow chart for preparing a compound of the present disclosure having a structure of Ring B according to one embodiment is shown;

[0140] Figure 5 A non-limiting flow chart for preparing a compound of the present disclosure having a structure of ring AB1 according to one embodiment is shown;

[0141] Figure 6 A non-limiting flow chart for preparing compounds of the present disclosure having a structure of ring AB2 according to one embodiment is shown;

[0142] Figure 7 A non-limiting scheme for preparing compounds of the present disclosure having a structure of ring AB3 according to one embodiment is shown;

[0143] Figure 8 A non-limiting scheme for preparing a compound of the present disclosure having a structure of ring ABCD1 according to one embodiment is shown;

[0144] Fig. 9 A non-limiting scheme for preparing compounds of the present disclosure having a structure of ring ABCD2 according to one embodiment is shown;

[0145] Fig.10 A non-limiting scheme for preparing compounds of the present disclosure having a structure of ring ABCD3 according to one embodiment is shown;

[0146] Fig.11 A non-limiting scheme for preparing a compound of the present disclosure according to formula (I) having a structure of ring ABCD1 according to one embodiment is shown;

[0147] Fig.12 A non-limiting scheme for preparing a compound of the present disclosure according to formula (III) having a structure of ring ABCD2 according to one embodiment is shown;

[0148] Fig.13 shows a non-limiting scheme for preparing a compound of the present disclosure according to formula (III) having a structure of ring ABCD3 according to one embodiment; and

[0149] Fig.14A non-limiting scheme for preparing a protected precursor of (2R,3R)-4-amino-3-fluoro-2-hydroxybutyric acid according to one embodiment is shown. DETAILED DESCRIPTION

[0150] The present disclosure will be described more fully below with reference to its exemplary embodiments. Before describing several exemplary embodiments of the present technology, it should be understood that the present technology is not limited to the details of the construction or process steps set forth in the following description. The present technology can have other embodiments and can be practiced or performed in various ways.

[0151] The following description sets forth many exemplary configurations, methods, parameters, etc., in order to provide a thorough understanding of various embodiments of the present disclosure. However, it should be appreciated that such description is not intended to limit the scope of the present disclosure, but is provided as a description of exemplary embodiments.

[0152] The present disclosure generally relates to aminoglycoside compounds, including stereoisomers, tautomers, solvates and pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising the compounds or stereoisomers, tautomers, solvates and pharmaceutically acceptable salts thereof. The present disclosure particularly relates to such compounds with antibacterial activity, stereoisomers, tautomers, solvates and pharmaceutically acceptable salts thereof. Therefore, compounds of the present disclosure and pharmaceutical compositions can be used for treating bacterial infections and medical conditions associated therewith. Compounds of the present disclosure and pharmaceutical compositions can also be used for treating refractory resistance mechanisms and multidrug resistant (MDR) bacterial infections.

[0153] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs.All patents and publications cited herein are incorporated by reference.

[0154] definition

[0155] Unless otherwise indicated, all scientific and technological terms used herein have the same common meaning as understood by those skilled in the art to which the present invention belongs. Regarding the terms used in this disclosure, the following definitions are provided. The application will use the following terms as defined below, unless the text context in which the term appears requires a different meaning.

[0156] The articles "a" and "an" used herein may refer to one or more than one (i.e., at least one) of the grammatical object of the article. For example, "an element" refers to one or more than one element.

[0157] The term "and / or" used in this disclosure may mean "and" or "or" unless otherwise stated.

[0158] The term "about" used throughout this specification is used to describe and explain small fluctuations. For example, the term "about" may refer to less than or equal to ±10%, or less than or equal to ±5%, such as 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%. Whether or not it is explicitly stated, all numerical values ​​herein are modified by the term "about". Values ​​modified by the term "about" certainly include specific values. For example, "about 5.0" necessarily includes 5.0.

[0159] As used herein, throughout the specification and claims, unless the context requires otherwise, the terms "including," "containing," and "comprising" are used in their open, non-limiting sense.

[0160] "Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, and preferably having 1 to 15 carbon atoms (i.e., C1-C 15 In certain embodiments, the alkyl group contains 1 to 13 carbon atoms (i.e., C1-C 13 In some embodiments, the alkyl group contains 1 to 8 carbon atoms (i.e., C1-C8 alkyl). In other embodiments, the alkyl group contains 1 to 5 carbon atoms (i.e., C1-C5 alkyl). In other embodiments, the alkyl group contains 1 to 4 carbon atoms (i.e., C1-C4 alkyl). In other embodiments, the alkyl group contains 1 to 3 carbon atoms (i.e., C1-C3 alkyl). In other embodiments, the alkyl group contains 1 to 2 carbon atoms (i.e., C1-C2 alkyl). In some embodiments, the alkyl group contains 2 carbon atoms (i.e., C2 alkyl or ethyl). In other embodiments, the alkyl group contains 1 carbon atom (i.e., C1 alkyl). In other embodiments, the alkyl group contains 5 to 15 carbon atoms (i.e., C5-C 15 In other embodiments, the alkyl group contains 5 to 8 carbon atoms (i.e., C5-C8 alkyl). In other embodiments, the alkyl group contains 2 to 5 carbon atoms (i.e., C2-C5 alkyl). In other embodiments, the alkyl group contains 3 to 5 carbon atoms (i.e., C3-C5 alkyl). In certain embodiments, the alkyl group is selected from the group consisting of: methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl group is attached to the remainder of the molecule by a single bond.

[0161] When used in conjunction with a chemical moiety (e.g., alkyl, alkenyl, or alkynyl), the term "C x-y" is intended to include groups containing from x to y carbons in the chain. For example, the term "C 1-6 "Alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight chain alkyl and branched chain alkyl groups containing 1 to 6 carbon atoms. The term -C x-y Alkylene- refers to a substituted or unsubstituted alkylene chain having from x to y carbons in the alkylene chain. For example, -C 1-6 Alkylene- may be selected from: methylene, ethylene, propylene, butylene, pentylene, and hexylene, any of which are optionally substituted.

[0162] "Alkoxy" refers to a group attached through an oxygen atom of the formula -O-alkyl wherein alkyl is an alkyl chain as defined above.

[0163] "Alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and preferably having 2 to 12 carbon atoms (i.e., C2-C 12 In some embodiments, the alkenyl group contains 2 to 8 carbon atoms (i.e., C2-C8 alkenyl). In some embodiments, the alkenyl group contains 2 to 6 carbon atoms (i.e., C2-C6 alkenyl). In other embodiments, the alkenyl group contains 2 to 4 carbon atoms (i.e., C2-C4 alkenyl). The alkenyl group is connected to the rest of the molecule by a single bond, such as vinyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, pent-1,4-dienyl, and the like.

[0164] "Alkynyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, and preferably having from 2 to 12 carbon atoms (i.e., C2-C 12 In some embodiments, the alkynyl group contains 2-8 carbon atoms (i.e., C2-C8 alkynyl). In other embodiments, the alkynyl group contains 2 to 6 carbon atoms (i.e., C2-C6 alkynyl). In other embodiments, the alkynyl group contains 2 to 4 carbon atoms (i.e., C2-C4 alkynyl). The alkynyl group is connected to the remainder of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc.

[0165] The term "C x-y "Alkenyl" and "C x-y "Alkynyl" refers to a substituted or unsubstituted unsaturated aliphatic group similar in length and possible substitution to the alkyl group described above, but containing at least one double bond or triple bond, respectively. The term -C x-y Alkenylene- refers to a substituted or unsubstituted alkenylene chain having from x to y carbons in the alkenylene chain. For example, -C 2-6Alkenylene-can be selected from: vinylene, propenylene, butenylene, pentenylene, and hexenylene, any of which are optionally substituted. The alkenylene chain can have 1 double bond or more than one double bond in the alkenylene chain. The term -C x-y Alkyne- refers to a substituted or unsubstituted alkynylene chain having from x to y carbons in the alkynylene chain. 2-6 Alkyne- may be selected from: ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any of which are optionally substituted. The alkynylene chain may have 1 triple bond or more than one triple bond in the alkynylene chain.

[0166] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the remainder of the molecule to a group, which consists only of carbon and hydrogen, does not contain unsaturation, and preferably has 1 to 12 carbon atoms, such as: methylene, ethylene, propylene, n-butylene, etc. The alkylene chain is connected to the remainder of the molecule and to the group by a single bond. The connection point of the alkylene chain to the remainder of the molecule and the connection point to the group can be achieved through any two carbons in the chain. In some embodiments, the alkylene contains 1 to 10 carbon atoms (i.e., C1-C8 alkylene). In some embodiments, the alkylene contains 1 to 8 carbon atoms (i.e., C1-C8 alkylene). In other embodiments, the alkylene contains 1 to 5 carbon atoms (i.e., C1-C5 alkylene). In other embodiments, the alkylene contains 1 to 4 carbon atoms (i.e., C1-C4 alkylene). In other embodiments, the alkylene contains 1 to 3 carbon atoms (i.e., C1-C3 alkylene). In other embodiments, the alkylene group contains 1 to 2 carbon atoms (i.e., C1-C2 alkylene). In other embodiments, the alkylene group contains 1 carbon atom (i.e., C1 alkylene). In other embodiments, the alkylene group contains 5 to 8 carbon atoms (i.e., C5-C8 alkylene). In other embodiments, the alkylene group contains 2 to 5 carbon atoms (i.e., C2-C5 alkylene). In other embodiments, the alkylene group contains 3 to 5 carbon atoms (i.e., C3-C5 alkylene).

[0167] "Alkenylene" or "alkenylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the remainder of the molecule to a radical, which consists only of carbon and hydrogen, contains at least one carbon-carbon double bond, and preferably has 2 to 12 carbon atoms. The alkenylene chain is connected to the remainder of the molecule and to the radical by a single bond. The point of attachment of the alkenylene chain to the remainder of the molecule and to the radical can be achieved through any two carbons in the chain. In certain embodiments, an alkenylene group contains 2 to 10 carbon atoms (i.e., C2-C 10In some embodiments, the alkenylene group contains 2 to 8 carbon atoms (i.e., C2-C8 alkenylene). In other embodiments, the alkenylene group contains 2 to 5 carbon atoms (i.e., C2-C5 alkenylene). In other embodiments, the alkenylene group contains 2 to 4 carbon atoms (i.e., C2-C4 alkenylene). In other embodiments, the alkenylene group contains 2 to 3 carbon atoms (i.e., C2-C3 alkenylene). In other embodiments, the alkenylene group contains 2 carbon atoms (i.e., C2 alkenylene). In other embodiments, the alkenylene group contains 5 to 8 carbon atoms (i.e., C5-C8 alkenylene). In other embodiments, the alkenylene group contains 3 to 5 carbon atoms (i.e., C3-C5 alkenylene).

[0168] "Alkynylidene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the remainder of the molecule to a radical, which consists only of carbon and hydrogen, contains at least one carbon-carbon triple bond, and preferably has 2 to 12 carbon atoms. The alkynylene chain is connected to the remainder of the molecule by a single bond and to the radical by a single bond. The point of attachment of the alkynylene chain to the remainder of the molecule and to the radical can be achieved through any two carbons in the chain. In certain embodiments, an alkynylene group comprises 2 to 10 carbon atoms (i.e., C2-C 10 In certain embodiments, the alkynylene group comprises 2 to 8 carbon atoms (i.e., C2-C8 alkynylene). In other embodiments, the alkynylene group comprises 2 to 5 carbon atoms (i.e., C2-C5 alkynylene). In other embodiments, the alkynylene group comprises 2 to 4 carbon atoms (i.e., C2-C4 alkynylene). In other embodiments, the alkynylene group comprises 2 to 3 carbon atoms (i.e., C2-C3 alkynylene). In other embodiments, the alkynylene group comprises 2 carbon atoms (i.e., C2 alkynylene). In other embodiments, the alkynylene group comprises 5 to 8 carbon atoms (i.e., C5-C8 alkynylene). In other embodiments, the alkynylene group comprises 3 to 5 carbon atoms (i.e., C3-C5 alkynylene).

[0169] "Aryl" refers to a radical derived from an aromatic monocyclic or polycyclic hydrocarbon ring system by removing hydrogen atoms from ring carbon atoms. The aromatic monocyclic or polycyclic hydrocarbon ring system contains only hydrogen and carbon atoms, and 5 to 18 carbon atoms, wherein at least one ring of the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) π-electron system conforming to the Hückel theory. Ring systems from which aromatic radicals are derived include, but are not limited to, radicals such as benzene, fluorene, indane, indene, tetralin, and naphthalene.

[0170] "Aralkyl" refers to a radical of the formula -R-aryl where R is an alkylene chain as defined above, eg, methylene, ethylene, and the like.

[0171] "Aralkenyl" refers to a radical of the formula -R-aryl, wherein R is an alkenylene chain as defined above. "Aralkynyl" refers to a radical of the formula -R-aryl, wherein R is an alkynylene chain as defined above.

[0172] "Carbocycle" refers to a saturated, unsaturated or aromatic ring in which each atom in the ring is carbon. Carbocycles 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 group. In some embodiments, the carbocycle is a cycloalkyl group. In some embodiments, the carbocycle is a cycloalkenyl group. 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). Where valence permits, any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocycle. Exemplary carbocycles include: cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl.

[0173] The term "cycloalkyl" refers to a stable, fully saturated monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms, including fused or bridged ring systems, and preferably having 3 to 12 carbon atoms. In certain embodiments, the cycloalkyl contains 3 to 10 carbon atoms. In other embodiments, the cycloalkyl contains 5 to 7 carbon atoms. The cycloalkyl can be connected to the remainder of the molecule by a single bond. Examples of monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyls include, for example, adamantyl, bornyl (i.e., bicyclo[2.2.1]heptyl), norbornenyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, etc.

[0174] The term "cycloalkenyl" refers to an unsaturated non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms, which comprises a fused or bridged ring system, preferably having 3 to 12 carbon atoms, and comprising at least one double bond. In certain embodiments, the cycloalkenyl comprises 3 to 10 carbon atoms. In other embodiments, the cycloalkenyl comprises 5 to 7 carbon atoms. The cycloalkenyl can be connected to the remainder in the molecule by a single bond. Examples of cycloalkenyl include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl.

[0175] "Cycloalkylalkyl" means a radical of the formula -R-cycloalkyl where R is an alkylene chain as described above.

[0176] "Cycloalkylalkoxy" refers to a group of the formula -OR c -cycloalkyl group bonded via an oxygen atom, wherein R c is an alkylene chain as described above.

[0177] "Halo" or "halogen" refers to halogen substituents such as bromo, chloro, fluoro, and iodo substituents.

[0178] As used herein, the term "haloalkyl" or "haloalkane" refers to an alkyl group as defined above substituted with one or more halogen groups, for example, trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. In some embodiments, the alkyl portion of the fluoroalkyl group is optionally further substituted. Examples of halogen-substituted alkanes ("haloalkanes") include halogenated methane (e.g., chloromethane, bromomethane, fluoromethane, iodomethane), dihalogenated and trihalogenated methane (e.g., chloroform, bromoform, trifluoromethane, iodoform), 1-haloethane, 2-haloethane, 1,2-dihaloethane, 1-halopropane, 2-halopropane, 3-halopropane, 1,2-dihalopropane, 1,3-dihalopropane, 2,3-dihalopropane, 1,2,3-trihalopropane, and any other suitable combination of an alkane (or substituted alkane) and a halogen (e.g., Cl, Br, F, I, etc.). If the alkyl group is substituted with more than one halogen group, each halogen may be independently selected, such as 1-chloro, 2-fluoroethane.

[0179] "Fluoroalkyl" refers to an alkyl group as defined above substituted with one or more fluorine groups, for example: trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like.

[0180] "Heterocycle" refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3 to 10-membered monocyclic rings, 6 to 12-membered bicyclic rings, and 6 to 12-membered bridged rings. Each ring in the bicyclic heterocycle can be selected from saturated, unsaturated, and aromatic rings. In some embodiments, the heterocycle is a heteroaryl. In some embodiments, the heterocycle is a heterocycloalkyl. "Heterocyclylene" refers to a divalent heterocycle that connects the remainder of the molecule to a radical.

[0181] "Heterocycloalkyl" refers to a stable 3 to 12-membered non-aromatic ring group containing 2 to 12 carbon atoms and at least one heteroatom, wherein each heteroatom can be selected from N, O, Si, P, B, and S atoms. The heterocycloalkyl can be selected from a monocyclic or bicyclic, and fused or bridged ring system. The heteroatoms in the heterocycloalkyl are optionally oxidized. One or more nitrogen atoms (if present) are optionally quaternized. The heterocycloalkyl is partially or completely saturated. Where valence permits, the heterocycloalkyl is connected to the remainder of the molecule through any atom in the heterocycloalkyl, such as any carbon atom or nitrogen atom of the heterocycloalkyl. Examples of heterocycloalkyl include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.

[0182] "Heterocycloalkylalkyl" refers to a radical of the formula -R-heterocycloalkyl, wherein R is an alkylene chain as defined above. If the heterocycloalkyl is a nitrogen-containing heterocycloalkyl, the heterocycloalkyl is optionally attached to the alkylene chain at the nitrogen atom.

[0183] "Heteroaryl" or "aromatic heterocycle" refers to a group derived from a 3- to 12-membered aromatic ring group containing 1 to 11 carbon atoms and at least 1 heteroatom, wherein each heteroatom can be selected from N, O, and S. As used herein, the heteroaryl ring can be selected from a monocyclic or bicyclic ring, and a fused bridged ring system ring, wherein at least one ring in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) π-electron system that conforms to the Hückel theory. The heteroatoms in the heteroaryl group may be optionally oxidized. One or more nitrogen atoms (if present) are optionally quaternized. Where valence permits, the heteroaryl group is connected to the rest of the molecule through any atom in the heteroaryl group, such as a carbon atom or a nitrogen atom of the heteroaryl group. Examples of heteroaryl groups include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolanyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolanyl, benzodioxinyl, benzo pyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothiophenyl, benzothieno[3,2d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2a]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-dihydro Benzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furano[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, indazole 1,2-dole, 1,2-dole-2-yl, 1,2-dole-3-yl, 1,2-dole-4-yl, 1,2-dole-5-yl, 1,2-dole-6-yl, 1,2-dole-7-yl, 1,2-dole-8-yl, 1,2-dole-9-yl, 1,2-dole-10-yl, 1,2-dole-20-yl, 1,2-dole-11-yl, 1,2-dole-21-yl, 1,2-dole-12-yl, 1,2-dole-13-yl, 1,2-dole-22-yl, 1,2-dole-14-yl, 1,2-dole-15-yl, 1,2-dole-16-yl, 1,2-dole-17-yl, 1,2-dole-18-yl, 1,2-dole-19-yl, 1,2-dole-19-yl, 1,2-dole-110a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl , quinazolinyl, quinoxalinyl, quinolinyl, isoquinoxalinyl, 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]pyridinyl, and thienyl (i.e., thienyl). "X-membered heteroaryl" refers to the number of ring atoms (i.e., X) in the ring. For example, a 5-membered heteroaryl ring or a 5-membered aromatic heterocyclic ring has 5 atoms in the ring, such as triazole, oxazole, thiophene, etc.

[0184] "Heteroarylalkyl" refers to a radical of the formula -R-heteroaryl, wherein R is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkylene chain at the nitrogen atom.

[0185] "Isomers" are different compounds with the same molecular formula. "Stereoisomers" are isomers that differ only in the way 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. Where appropriate, the term "(±)" is used to represent a racemic mixture. "Diastereoisomers" or "diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified by R or S. Compounds whose absolute configuration is unknown can be specified as (+) or (-) according to the direction (right-handed or left-handed) in which they rotate the plane polarized light of the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms, whose asymmetric centers may be defined in terms of absolute stereochemistry as (R)- or (S)-. The chemical entities, pharmaceutical compositions, and methods of the present invention are intended to include all such possible stereoisomers, including racemic mixtures, optically pure forms, diastereomeric mixtures, and intermediate mixtures. Optically active (R)- and (S)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. The optical activity of the compounds may be analyzed by any suitable method, including, but not limited to, chiral chromatography and polarimetry, and the degree of predominance of one stereoisomer over another may be determined.

[0186] In certain embodiments, the compounds of the present disclosure may contain asymmetric or chiral centers and therefore exist in different stereoisomeric forms. The term "stereoisomer" may refer to a group of compounds that have the same number and type of atoms, the same bonds between these atoms, but the compounds are different in three-dimensional structure. The term "stereoisomer" may refer to any member of the group of compounds. For example, a stereoisomer may be an enantiomer or a diastereomer. It is intended that all stereoisomeric forms of the compounds of the present disclosure and mixtures thereof (including racemic mixtures) constitute part of the present disclosure.

[0187] When stereochemistry is not specified, some molecules described herein include isomers, such as enantiomers and diastereomers, mixtures of enantiomers, including racemates, mixtures of diastereomers, and other mixtures thereof, as long as they can be made by those of ordinary skill in the art by routine experiments. In some embodiments, single enantiomers or diastereomers (i.e., optically active forms) can be obtained by asymmetric synthesis or by splitting a mixture of racemates or diastereomers. If possible, the splitting of a racemate or a mixture of diastereomers can be completed, for example, by conventional methods, such as crystallization in the presence of a resolving agent, or using the chromatography of a chiral high pressure liquid chromatography (HPLC) column, for example. In addition, a mixture of two enantiomers (one of which is rich in both) can be purified by recrystallization and / or grinding (trituration), so as to provide a further optically enriched form of a major enantiomer.

[0188] In certain embodiments, the chiral centers of the present disclosure can have the S or R configuration as defined in the IUPAC 1974 recommendations.

[0189] As used herein, the term "amino" refers to -NH2.

[0190] As used herein, the term "hydroxy" or "hydroxyl" refers to -OH.

[0191] As used herein, the term "oxo" refers to a "=O" group. It may also be abbreviated as C(O) or C=O in the text.

[0192] The term "substituted" refers to a substituent in a part that replaces one or more carbons in the structure or hydrogen on a heteroatom (e.g., NH) that can be substituted. It should be understood that the implicit condition included in "replacement" or "replacement with..." is that this substitution conforms to the valence allowed by the substituted atom and the substituent, and the substitution results in a stable compound, i.e., the compound will not spontaneously transform by, for example, rearrangement, cyclization, elimination, etc. In certain embodiments, substitution refers to a substituent in a part that replaces two hydrogen atoms on the same carbon atom, such as replacing two hydrogen atoms on a single carbon with an oxo, imino, or thio group. As used herein, the term "substituted" is intended to include all allowed substituents of an organic compound. In a broad sense, allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. For appropriate organic compounds, allowed substituents may be one or more and the same or different. For the purposes of this disclosure, heteroatoms (such as nitrogen) may have hydrogen substituents, and / or any allowed substituents of organic compounds described herein that satisfy the valence of heteroatoms.

[0193] In some embodiments, the substituents may include any of the substituents described herein, for example, halogen, hydroxy, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=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 -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2), and -R b -S(O) t N(R a )2 (wherein 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 (=NH), oxime (=N-OH), hydrazine (=N-NH2), -R b -OR a , -Rb -OC(O)-R a , -R b -OC(O)-OR a , -R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2), and -R b -S(O) t N(R a )2 (where t is 1 or 2); where each R a are independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, wherein each R a Where valence permits, it may be optionally substituted by: alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=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(Ra )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2), and -R b -S(O) t N(R a )2 (where t is 1 or 2); and wherein each R b are independently selected from a direct bond, or a straight or branched alkylene, alkenylene, or alkynylene chain, and each R c Each is a straight or branched alkylene, alkenylene, or alkynylene chain.

[0194] As used herein, the term "group of a compound" refers to a structure derived from a parent compound by removing one or more atoms (e.g., hydrogen atoms). In one embodiment, a "group of a compound" is a monovalent group derived by removing a hydrogen atom from a parent compound.

[0195] It will be understood that certain group naming conventions may include monoradicals or diradicals, depending on the context. For example, if a substituent requires two points of attachment to the rest of the molecule, it is understood that the substituent is a diradical. For example, substituents defined as alkyl groups that require two points of attachment include diradicals such as: -CH2-, -CH2CH2-, -CH2CH(CH3)CH2-, and the like. Other group naming conventions explicitly indicate that the group is a diradical, such as "alkylene," "alkenylene," "arylene," and the like.

[0196] Whenever a substituent is described as a diradical (ie, having two points of attachment to the rest of the molecule), it is understood that the substituent may be attached in any orientation configuration unless otherwise specified.

[0197] "Tautomers" refer to molecules in which a proton moves from one atom in a molecule to another atom in the molecule. In certain embodiments, the compounds presented herein exist as tautomers. In situations 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:

[0198]

[0199] As used herein, the term "unsubstituted" may mean that a particular group bears no substituents other than the moiety being described (eg, where hydrogen satisfies valencies).

[0200] "Stable compound" and "stable structure" may refer to a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0201] As used herein, the terms "parenteral administration" and "administered parenterally" refer to forms of administration / administration other than enteral and topical administration, usually by injection, including but not limited to intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraocular, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion administration / administration.

[0202] The phrase "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions and / or dosage forms that are, in accordance with a reasonable benefit / risk ratio, suitable for use in contact with the tissues of humans and animals without causing excessive toxicity, irritation, allergic response, or other problems or complications according to sound medical judgment.

[0203] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; (4) powdered gum tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0204] The term "carrier" used in the present disclosure may include carriers, excipients and diluents, and may refer to materials, compositions or carriers that are involved in carrying or transporting a medicament from one organ or part of a subject's body to another organ or part, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, wherein the medicament is, for example, one or more compounds of the present disclosure, or a pharmaceutically acceptable salt, solvate (e.g., hydrate), isomer (e.g., stereoisomer) and tautomer thereof. The carrier should be selected based on the compatibility and release profile properties of the desired dosage form. Exemplary carrier materials may include, for example, adjuvants, adhesives, suspending agents, disintegrants, fillers, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, spray-dried dispersions, etc. See, for example, Hoover, John E. "Remington's Pharmaceutical Sciences", 18th edition, (1990, Mack Publishing Company, Easton, Pennsylvania 1975). Exemplary carrier materials may also include, but are not limited to, any adjuvant, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier certified by the U.S. Food and Drug Administration as acceptable for use in humans or domestic animals.

[0205] The term "pharmaceutically acceptable" or "pharmacologically acceptable" may refer to a material that is not biologically or otherwise undesirable - the material may be administered to a subject without causing any substantial undesirable biological effect or interacting in a deleterious manner with any ingredients of a composition in which the material is contained.

[0206] "Pharmaceutical composition" may refer to a preparation of a compound of the present disclosure and a medium generally accepted in the art for delivering a biologically active compound to a subject (eg, a mammal or a human). Such a medium may include all pharmaceutically acceptable carriers therefor.

[0207] The terms "subject," "individual," and "patient" are used interchangeably and refer to humans as well as non-human mammals (e.g., non-human primates, canines, equines, felines, porcines, bovines, ungulates, lagomorphs, etc.). In various embodiments, the subject may be a human (e.g., an adult male, an adult female, an adolescent male, an adolescent female, a male child, a female child) who is under the care of a physician or other health worker in a hospital, as an outpatient clinic, or other clinical setting. In certain embodiments, the subject may not be under the care or prescription of a physician or other health worker.

[0208] As used herein, the phrase "a subject in need thereof" refers to a subject suffering from or at risk of a pathology, as described below, who is to be treated prophylactically or therapeutically with a compound or salt described herein.

[0209] "Agent" or "bioactive agent" as used herein refers to an organism, drug or compound or other part. Non-limiting examples include organic or inorganic molecules, peptides, proteins, peptide nucleic acids (PNA), oligonucleotides (including, for example, aptamers and polynucleotides), antibodies, antibody derivatives, antibody fragments, vitamin derivatives, carbohydrates, toxins, branched-chain amino acids in the form of free amino acids or their metabolites, or chemotherapy (chemotherapeutic) compounds. Various compounds can be synthesized, such as small molecules and oligomers (such as 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, etc. Those skilled in the art can easily appreciate that the structural properties of the agents are unlimited.

[0210] The term "administer, administered, administrators, administering" is defined as providing a composition to a subject by a route known in the art, including but not limited to intravenous, intraarterial, oral, parenteral, oral, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal or intraperitoneal administration routes. In certain embodiments, an oral route of administration of the composition may be used. The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or salt described herein sufficient to affect the intended application (including but not limited to disease treatment as defined below). The therapeutically effective amount may vary depending on the intended application (in vitro or in vivo), or the subject and disease condition being treated, for example, the subject's weight and age, the severity of the disease condition, the mode of administration, etc., which can be easily determined by a person of ordinary skill in the art. The term also applies to doses that can induce a specific response in a target cell, for example, a decrease in proliferation or downregulation of activity of a target protein. The specific dosage may vary depending on the specific compound selected, the dosing regimen to be followed, whether it is administered in conjunction with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system that carries it.

[0211] As used herein, "treatment" or "treatment" refers to a method of obtaining a beneficial or desired result associated with a disease, disorder or medical condition, including but not limited to a therapeutic benefit and / or a preventive benefit. In certain embodiments, treatment or treatment involves administering a compound or composition disclosed herein to a subject. A therapeutic benefit may include eradication or improvement of the underlying disorder being treated. In addition, a therapeutic benefit may be achieved by eradication or improvement of one or more physiological symptoms associated with the underlying disorder, such as an improvement observed in a subject, although the subject may still be suffering from the underlying disorder. In certain embodiments, for preventive benefit, the composition may be administered to a subject at risk of developing a particular disease or to a subject reporting one or more physiological symptoms of a disease, even if a diagnosis of the disease is not possible. Treatment may include, for example, reducing, delaying or alleviating the severity of one or more symptoms of a disease or disorder, or it may include reducing the frequency with which a patient experiences symptoms of a disease, defect, disorder or adverse condition. As used herein, treatment may be used to refer to a method that results in treatment or improvement of a disease or disorder at some level, and a range of results for this purpose may be considered, including but not limited to preventing the disorder completely.

[0212] In certain embodiments, the term "prevent" or "preventing" in relation to a disease or disorder may refer to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in a treated sample 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 an untreated control sample.

[0213] The term "prodrug" is intended to encompass compounds that are converted to the therapeutic agents of the present disclosure under physiological conditions. A common method of preparing a prodrug is to include one or more selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids, and phosphonates) are preferred prodrugs of the present disclosure.

[0214] As used herein, the term "treat / treat" can include alleviating, alleviating or ameliorating symptoms of a disease or condition, preventing additional symptoms, ameliorating or preventing the underlying cause of symptoms, inhibiting the disease or condition, such as preventing the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, alleviating conditions caused by the disease or condition, or preventing the symptoms of the disease or condition prophylactically and / or therapeutically.

[0215] The term "effective amount" or "therapeutically effective amount" when used with one or more compounds or pharmaceutical compositions, may refer to an amount of one or more compounds or pharmaceutical compositions sufficient to provide a desired biological result. The result may be a reduction and / or alleviation of the signs, symptoms or causes of a condition, or any other desired change in a biological system. For example, an "effective amount" for therapeutic use may be the amount of a pharmaceutical composition as disclosed herein, comprising one or more compounds or pharmaceutically acceptable salts thereof, solvates (e.g., hydrates), isomers (e.g., stereoisomers), and tautomers, required to provide a clinically significant reduction in the condition. In any individual case, an appropriate "effective amount" may be determined by a person of ordinary skill in the art using routine experimentation.

[0216] In certain embodiments, the terms "disease" and "disorder" may be used interchangeably, or may be different, in the latter case because a particular disease or disorder may not have a known causative agent (and thus the cause is not yet known) and therefore is not yet considered a disease but rather merely an undesirable condition or syndrome in which a more or less specific set of symptoms has been identified by a clinician.

[0217] The terms "antimicrobial agent," "antibiotic," or "antibacterial compound" are used interchangeably herein and may refer to an agent or compound having bactericidal or bacteriostatic activity. The term "inhibiting growth" may mean that the rate of increase in the number of a particular bacterial population is reduced. Thus, the term may include situations where the bacterial population increases but at a reduced rate, as well as situations where the growth of the bacterial population is stopped, as well as situations where the number of bacteria in the bacterial population is reduced or even eliminated. The activity of an antimicrobial agent or compound is not necessarily limited to bacteria, but may also encompass general antimicrobial activity against parasites or fungi, or general antiviral activity against viruses.

[0218] Compounds of the Disclosure

[0219] In general, the present disclosure provides aminoglycoside compounds that can be characterized as having antibacterial activity.

[0220] In one aspect, provided are compounds having a structure according to Formula (I):

[0221]

[0222] or a pharmaceutically acceptable salt or solvate thereof;

[0223] in:

[0224] R a is hydrogen (H) or methyl; and

[0225] R b C 1-6Alkyl, or having a structure according to formula (II):

[0226]

[0227] in:

[0228] Q is NH, O or S;

[0229] n is an integer from 0 to 4;

[0230] R 1 is hydrogen or C 1-3 alkyl;

[0231] R 2 and R 3 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, halogen, and -OH; and

[0232] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b Each independently selected from H and C 1-3 Alkyl; or

[0233] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0234] In another aspect, provided are compounds having a structure according to formula (III):

[0235]

[0236] or a pharmaceutically acceptable salt or solvate thereof;

[0237] in:

[0238] R a is H or methyl; and

[0239] R b H, C 1-6 Alkyl, or having a structure according to formula (II):

[0240]

[0241] in:

[0242] Q is NH, O or S;

[0243] n is an integer from 0 to 4;

[0244] R1 is hydrogen or C 1-3 alkyl;

[0245] R 2 and R 3 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, halogen, and -OH; and

[0246] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b are each independently selected from hydrogen and C 1-3 Alkyl; or

[0247] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0248] Those skilled in the art will appreciate that the compounds of the present disclosure according to formula (I) and formula (III) all have a common hexahydropyranyl ring system:

[0249]

[0250] wherein Z is H (formula (I)) or methyl (formula (III)).

[0251] Without wishing to be bound by any particular theory, it is believed that the specific hexahydropyranyl ring system of compounds according to formula (I) and formula (III) can impart favorable antibacterial activity to compounds containing such ring systems. The compounds of formula (I) and formula (III) further contain additional optionally substituted saturated carbocyclic or heterocyclic ring systems (e.g., tricyclic ring systems, such that the compounds of formula (I) and formula (III) each contain four connected rings including the hexahydropyranyl ring shown), which together constitute the aminoglycoside core. In this article, with reference to the tetracyclic ring system (aminoglycoside) that constitutes the compound, the compounds of formula (I) and formula (III) may be referred to as "ABCD compounds", wherein the A, B, C and D rings are as follows:

[0252]

[0253] Herein, conventional numbering is applied to such ring systems. For simplicity, the conventional numbering is described below with reference to a simplified, non-limiting ABCD ring skeleton (ABCD', i.e., many substituents have been removed relative to formula (I) and formula (III) for ease of visualization):

[0254]

[0255] The structure of the ABCD' ring backbone may alternatively be described in three-dimensional form as shown below:

[0256]

[0257] in:

[0258] In some embodiments, provided are compounds having a structure according to formula (I),

[0259]

[0260] in:

[0261] R a is hydrogen (H) or methyl; and

[0262] R b C 1-6 Alkyl, or having a structure according to formula (II):

[0263]

[0264] in:

[0265] Q is NH, O or S;

[0266] n is an integer from 0 to 4;

[0267] R 1 is hydrogen or C 1-3 alkyl;

[0268] R 2 and R 3 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, halogen, and -OH; and

[0269] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b Each independently selected from H and C 1-3 Alkyl; or

[0270] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0271] In some embodiments, R a For H.

[0272] In some embodiments, R a It is methyl (CH3).

[0273] In some embodiments, R b C 1-6 Alkyl, such as methyl, ethyl, propyl, butyl (including isobutyl and sec-butyl), pentyl (including all possible isomers thereof), or hexyl (including all possible isomers thereof).

[0274] In some embodiments, R b It is ethyl (C2H5).

[0275] In some embodiments, R b is ethyl, and the compound of formula (I) has the following structure:

[0276]

[0277] In some embodiments, R b is ethyl, and the compound of formula (I) has the following structure:

[0278]

[0279] In some embodiments, R b has a structure according to formula (II) as defined above.

[0280] In some embodiments, referring to formula (II), Q is NH; n is an integer from 1 to 3; R 1 is hydrogen; R 2 and R 3 Each occurrence is independently selected from hydrogen, alkyl, halogen, and -OH; and R 4 is H. In some embodiments, n is 2.

[0281] In some embodiments, R b for Wherein n is 1, 2, or 3.

[0282] In some embodiments, R b Selected from:

[0283]

[0284] In some embodiments, R b Selected from:

[0285]

[0286] In some embodiments, the compound of formula (I) has a structure selected from the following:

[0287] In some embodiments, the compound of formula (I) has the following structure:

[0288] In some embodiments, the compound of formula (I) has the following structure:

[0289] In some embodiments, the compound of formula (I) has a structure selected from the following:

[0290] In some embodiments, the compound of formula (I) has the following structure:

[0291]

[0292] In some embodiments, the compound of formula (I) has the following structure:

[0293] In another aspect, provided are compounds having a structure according to formula (III):

[0294] in:

[0295] R a is H or methyl; and

[0296] R b H, C 1-6 Alkyl, or having a structure according to formula (II):

[0297] in:

[0298] Q is NH, O or S;

[0299] n is an integer from 0 to 4;

[0300] R 1 is hydrogen or C 1-3 alkyl;

[0301] R 2 and R 3 Each occurrence is independently selected from: hydrogen, alkyl, halogen, and -OH; and

[0302] R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b are each independently selected from hydrogen and C 1-3 Alkyl; or

[0303] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0304] In some embodiments, R a For H.

[0305] In some embodiments, R a It is methyl.

[0306] In some embodiments, the compound of formula (III) has a structure selected from the following:

[0307]

[0308] In some embodiments, R b For H.

[0309] In some embodiments, R b C 1-6 Alkyl, such as methyl, ethyl, propyl, butyl (including isobutyl and sec-butyl), pentyl (including all possible isomers thereof), or hexyl (including all possible isomers thereof). b It is ethyl.

[0310] In some embodiments, R b is ethyl, and the compound of formula (I) has a structure selected from the following:

[0311]

[0312] In some embodiments, R b has a structure according to formula (II) as defined above.

[0313] In some embodiments, referring to formula (II), Q is NH; n is an integer from 1 to 3; R 1 is hydrogen; R 2 and R 3 Each occurrence is independently selected from hydrogen, alkyl, halogen, and -OH; and R 4 is H. In some embodiments, n is 2.

[0314] In some embodiments, R b for Wherein n is 1, 2, or 3.

[0315] In some embodiments, R b Selected from:

[0316]

[0317] In some embodiments, R b Selected from:

[0318]

[0319] In some embodiments, the compound of formula (III) has a structure selected from the following:

[0320]

[0321] In some embodiments, the compound of formula (III) has the following structure:

[0322]

[0323] In some embodiments, the compound of formula (III) has the following structure:

[0324]

[0325] In some embodiments, the compound of formula (III) has the following structure:

[0326] In some embodiments, the compound of formula (III) has the following structure:

[0327]

[0328] In some embodiments, the compound of formula (III) has the following structure:

[0329]

[0330] In some embodiments, the compound of formula (III) has the following structure:

[0331]

[0332] In some embodiments, the compound of formula (III) has the following structure:

[0333]

[0334] In some embodiments, the compound of formula (III) has the following structure:

[0335]

[0336] Isotopes and isotope-labeled compounds

[0337] The compounds described herein may exhibit their natural isotopic abundance, or one or more atoms may be artificially enriched in a particular isotope having the same atomic number but an atomic mass or mass number different from that predominantly found in nature. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure. Thus, reference to an element is intended to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, then the group also includes isotopes of hydrogen. For example, hydrogen has three naturally occurring isotopes, represented as 1 H (hydrogen), 2 H (deuterium), and 3 H (tritium). Protium is the most abundant isotope of hydrogen in nature. Deuterium enrichment can provide certain therapeutic advantages, such as increasing half-life and / or exposure in vivo, or can provide compounds that can be used to study in vivo pathways of drug elimination and metabolism. Isotopically enriched compounds can be prepared by conventional techniques well known to those skilled in the art.

[0338] The compounds described herein further include all pharmaceutically acceptable isotope-labeled compounds. An "isotope-labeled" or "radiolabeled" compound may be a compound in which one or more atoms are replaced or substituted by an atom with an atomic mass or mass number different from that normally present in nature (i.e., naturally occurring). For example, in some embodiments, in the compounds described herein, a hydrogen atom is replaced or substituted by one or more deuterium or tritium.

[0339] Certain isotope-labeled compounds of the present disclosure (e.g., compounds that include a radioactive isotope) can be used in drug and / or substrate tissue distribution studies. 3 H, and carbon 14, i.e. 14 C, are particularly useful for this purpose because they are easy to incorporate and detection methods are readily available. 2 H) substitution may provide certain therapeutic advantages due to higher metabolic stability, such as increasing half-life in vivo or reducing dosage requirements, and may therefore be preferred in some cases. In some embodiments, the compound comprises at least one deuterium atom. For example, in the compounds of the present disclosure, one or more hydrogen atoms may 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 into the compounds described herein include, but are not limited to: 2 H (also written as D, representing deuterium), 3 H (also written as T, representing tritium), 11 C.13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 18 F. 35 S. 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I. 124 I. 125 I, and 131 I. Using positron-emitting isotopes (such as 11 C. 18 F. 15 O. and 13 N) substitution may be useful in Positron Emission Topography (PET).

[0340] Isotopically labeled forms 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 labeled reagent for a non-isotopically labeled reagent.

[0341] The disclosure herein is also intended to cover the in vivo metabolites of the disclosed compounds. Such products can be obtained by oxidation, reduction, hydrolysis, amidation, esterification, etc. of the administered compound, mainly due to enzymatic processes. Therefore, the disclosure may include compounds produced by the following method: the method includes administering the compound of the present disclosure to a subject, such as a mammal, for a period of time sufficient to obtain the metabolites of the subject. Such products are generally identified as follows: administering the disclosed radiolabeled compound to a subject (e.g., rat, mouse, guinea pig, monkey or human) at a detectable dose for a period of time long enough for metabolism to occur, and isolating its conversion products from urine, blood or other biological samples.

[0342] Salts and solvates

[0343] The present disclosure further provides pharmaceutically acceptable salts, solvates (e.g., hydrates), and combinations thereof of any of the compounds disclosed herein. The use of the terms "salt," "hydrate," "solvate," and the like is intended to be equally applicable to salts, hydrates, or solvates of enantiomers, diastereomers, isomers, stereoisomers, rotational isomers, tautomers, positional isomers, or racemates of the disclosed compounds.

[0344] The term "salt" or "pharmaceutically acceptable salt" refers to salts derived from a variety of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed from inorganic and organic acids. Inorganic acids from which salts can be obtained include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Organic acids from which salts can be obtained 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, etc. Pharmaceutically acceptable base addition salts can be formed from inorganic and organic bases. Inorganic bases from which salts can be obtained include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Organic bases from which salts can be obtained 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 selected from ammonium, potassium, sodium, calcium and magnesium salts. In some embodiments, "pharmaceutically acceptable salts" may include, for example, water-soluble salts and water-insoluble salts, such as acetate, stilbenesulfonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camphorsulfonate, carbonate, chloride, citrate, clavulanate, dihydrochloride, edetate, edisylate, propionate lauryl sulfate, ethanesulfonate, fumarate, glucoheptonate, gluconate, glutamate, glycolyl arsanoate, hexafluorophosphate, hexylresorcinate, halamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate , lactate, lactobionate, laurate, magnesium, malate, maleate, mandelate, methanesulfonate, methyl bromide, methyl nitrate, methyl sulfate, mucate, naphthenate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate, 1,1-methylene-bis-2-hydroxy-3-naphthoate, pamoate, einbonate, pantothenate, phosphate / diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate, sulfamate, tannate, tartrate, theoclate, toluenesulfonate, triethyl iodide, and valerate.

[0345] The compounds of the present invention also include crystalline and amorphous forms of these compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same activity, including, for example: polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs and amorphous forms of the compounds, and mixtures thereof.

[0346] The compounds of the present disclosure (including stereoisomers and tautomers thereof, and salts of any of them) may exist as solvates. Typically, crystallization produces solvates of the compounds of the present disclosure. The term "solvate" as used herein refers to: an aggregate comprising one or more molecules of a solvent having one or more molecules and a compound of the present disclosure. The solvent may be water, in which case the solvate may be a hydrate. Alternatively, the solvent may be an organic solvent. Therefore, the compounds of the present disclosure and salts may exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvate forms. The compounds of the present disclosure may be true solvates, while in other cases, the compounds of the present disclosure may retain only exogenous water or a mixture of water and some exogenous solvents.

[0347] Preparation of compounds

[0348] Compounds of the present disclosure having an ABCD ring system as described above (e.g., compounds of formula (I) and formula (III)) can be prepared by methods known in the art of organic synthesis, which are partially described by the following description, examples, and synthetic schemes in conjunction with the guidance provided herein. In the schemes described below and provided in the figures, it is understood that, if necessary, protecting groups can be used for sensitive or reactive groups according to the guidance provided herein, according to general principles or chemistry. Protecting groups can be operated according to standard methods of organic synthesis (TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis", 3rd edition, Wiley, New York, 1999). These groups can be removed at a convenient stage of compound synthesis using methods that are clear to those skilled in the art based on the detailed teachings provided herein. The selection process and reaction conditions and the order in which they are performed should be consistent with the present disclosure.

[0349] In general, the preparation of the disclosed compounds having an ABCD ring system comprises a combination of reactions and conditions that provide certain novel intermediate compounds. Figure 1 Representative, non-limiting strategies for preparing compounds having an ABCD ring system (eg, compounds of Formula (I) and Formula (III)) are described.

[0350] Solution 1

[0351]

[0352] Aspects of the preparation methods according to Scheme 1 are discussed further below. The reactions in Scheme 1 can be carried out by 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.

[0353] Preparation of A-ring compounds

[0354] According to Scheme 1, an exemplary preparation of compounds of the present disclosure having an ABCD ring system (e.g., compounds of Formula (I) and Formula (III)) uses an A ring compound. In some embodiments, the A ring compound has the general structure (A):

[0355]

[0356] in:

[0357] X 1 is an amine, an amine precursor, or a protected amino group;

[0358] X 2 is an amine, an amine precursor, or a protected amino group; and

[0359] R 5 It is hydrogen (H) or methyl (CH3).

[0360] In some embodiments, R 5 It's H.

[0361] In some embodiments, R 5 For CH3.

[0362] In some embodiments, X 1 is azide (N3). In some embodiments, X 1 In some embodiments, X 1 It is -N(CH3)(Cbz).

[0363] In some embodiments, X 2 It is azide (N3).

[0364] In some embodiments, the A ring compound has a structure of formula A1 (ie, Ring A1):

[0365]

[0366] Figure 1Exemplary schemes for preparing Ring A1 are illustrated and discussed in more detail herein. Generally, Ring A1 can be prepared starting from 2-formyl-3,4-dihydro-2H-pyran using a combination of condensation, reduction, oxidation, alkylation, acylation, deprotection, and substitution reactions.

[0367] Reference Figure 1 In some embodiments, 2-formyl-3,4-dihydro-2H-pyran is converted to a chiral sulfinylimine in the presence of a Lewis acid dehydrating agent such as copper sulfate, such as in the case of (S)-2-methylpropane-2-sulfinamide. The sulfinylimine can then be reduced to the corresponding sulfinamide using a suitable reducing agent such as sodium borohydride. In some embodiments, the nitrogen of the resulting sulfinamide is then protected using a suitable protecting group such as benzyl. In some embodiments, tetrahydropyran is converted to the corresponding halogenated lactal by reaction with, for example, bromine or iodine under aqueous conditions. In some embodiments, the hydroxyl group of the halogenated lactal is oxidized using a suitable oxidizing agent such as a hexavalent chromium-based reagent to form the corresponding lactone. In some embodiments, the azide group is introduced by replacing the halogen (e.g., iodine) group with an azide ion source. In some embodiments, the lactone carbonyl is reduced using a suitable reducing agent such as an aluminum hydride reagent to form the corresponding lactone. Subsequently, the sulfenamide chiral auxiliary can be cleaved, for example under acidic conditions, and the resulting amino group protected with a suitable protecting group (eg benzyl carbamate) to afford Ring A1.

[0368] In some embodiments, the A ring compound has a structure of formula A2 (ie, Ring A2):

[0369]

[0370] An exemplary scheme for preparing ring A2 is as follows Figure 2 As shown in . Figure 2In some embodiments, in the presence of a Lewis acid dehydrating agent (e.g., copper sulfate), the aldehyde is converted into a chiral sulfenyl imine, such as in the case of (S)-2-methylpropane-2-sulfenamide. Subsequently, the sulfenyl imine can be treated with a suitable organometallic reagent (e.g., methylmagnesium bromide) to obtain a chiral sulfenamide. In some embodiments, a suitable protecting group (e.g., benzyl) is then used to protect the nitrogen of the resulting sulfenamide. In some embodiments, tetrahydropyran is converted into a corresponding halogenated lactal by reacting with, for example, bromine or iodine under aqueous conditions. In some embodiments, the hydroxyl group of the halogenated lactal is oxidized using a suitable oxidant (e.g., a reagent based on hexavalent chromium) to form a corresponding lactone. In some embodiments, a halogen (e.g., iodine) group is replaced with an azide ion source, thereby introducing an azide group. In some embodiments, a suitable reducing agent (e.g., aluminum hydride reagent) is used to reduce the lactone carbonyl to form a corresponding lactal. The sulfenamide chiral auxiliary can then be cleaved, for example under acidic conditions, and the resulting amino group protected with a suitable protecting group (eg, benzyl carbamate) to provide Ring A2.

[0371] In some embodiments, the A ring compound has a structure of formula A3 (ie, Ring A3):

[0372]

[0373] Ring A3 can be based on Figure 2 Prepared as described in the general protocol, but using Figure 3 The R-enantiomer of 2-methylpropane-2-sulfenamide is shown in FIG.

[0374] In some embodiments, the A ring compound has a structure of formula A4 (ie, Ring A4):

[0375]

[0376] In some embodiments, the A ring compound has a structure of formula A5 (ie, Ring A5):

[0377]

[0378] Ring A4 and Ring A5 can be respectively Figure 2 and Figure 3 Ring A2 and Ring A3 were prepared according to the general scheme discussed, but replacing the benzyl protecting group with a methyl group.

[0379] Preparation of B-ring compounds

[0380] According to Scheme 1, an exemplary preparation of compounds of the present disclosure having an ABCD ring system (e.g., compounds of Formula (I) and Formula (III)) uses a B ring compound. In some embodiments, the B ring compound has the following general structure:

[0381]

[0382] in:

[0383] X is an amine, an amine precursor, or a protected amino group; and

[0384] R is a protecting group.

[0385] In some embodiments, X is NH(Cbz) and R is Ac. Therefore, in some embodiments, the B ring compound has a structure of formula B1 (ie, ring B1):

[0386]

[0387] An exemplary method for preparing ring B1 is as follows Figure 4 See Figure 4 , generally, ring B1 can be prepared from dihydrostreptomycin by hydrolysis of the guanidine group, N-protection, O-protection, and hydrolysis of the glycosyl bond. Procedures for preparing such compounds are reported, for example, in International Patent Application Publication Nos. WO2019194858 and WO2019046126, both of which are attributed to Achaogen, Inc., each of which is incorporated herein by reference for the teaching of the synthesis of ring B1.

[0388] Preparation of AB ring compounds

[0389] According to Scheme 1, an exemplary preparation of a disclosed compound having an ABCD ring system (e.g., a compound of Formula (I) and Formula (III)) uses an AB ring compound. In general, referring to Scheme 1, the A ring and the B ring compounds are linked via a glycosidic bond to form an AB ring compound. Typically, the reaction is carried out under conditions that allow chemical glycosylation by coupling a glycosyl donor and a glycosyl acceptor. For example, a hydroxyl group on the B ring compound can act as a glycosyl donor and react with an A ring compound having a suitable leaving group to act as a glycosyl acceptor.

[0390] As a non-limiting example, in a suitable solvent, the anomeric hydroxyl group of the A ring compound is converted into a suitable leaving group, which can react with the hydroxyl group on the B ring compound to form an interglycosidic bond (interglycosidic linkage), and is allowed to react with the free hydroxyl group on the B ring compound to form the bond. Examples of suitable leaving groups include, but are not limited to, halides, alkylthioates, arylthioates, imidoesters / salts, acetates / salts, phosphates / salts, O-pentenyls, mesylates / salts, toluenesulfonates / salts, etc. In one embodiment, the A ring compound is treated with an activator (e.g., trichloroacetonitrile) to form an imidoester, and the B ring compound is added, followed by an additional activator (e.g., Lewis acid). In some embodiments, the Lewis acid is trimethylsilyl trifluoromethanesulfonate / ester, trimethylaluminum, or boron trifluoride etherate.

[0391] The structure of the AB ring compound can vary. In some embodiments, the AB ring compound has the following general structure:

[0392]

[0393] in:

[0394] X 1 , X 2 , and X 3 is in each instance independently selected from amines, amine precursors, and protected amino groups;

[0395] R 5 is H or CH3; and

[0396] R 6 and R 7 is in each instance independently selected from H, Bn and Piv.

[0397] In some embodiments, R 5 It's H.

[0398] In some embodiments, R 5 For CH3.

[0399] In some embodiments, X 1 is azide (N3). In some embodiments, X 1 In some embodiments, X 1 In some embodiments, X 1 It is -N(CH3)(Cbz).

[0400] In some embodiments, X 2 is azide (N3). In some embodiments, X2 is -NH(Cbz).

[0401] In some embodiments, X 3 is azide (N3). In some embodiments, X 3 is -NH(Cbz).

[0402] In some embodiments, R 6 is benzyl. In some embodiments, R 6 It is pivaloyl.

[0403] In some embodiments, R7 is H.

[0404] In some embodiments, the AB ring compound has a structure selected from the following:

[0405]

[0406] In some embodiments, the AB ring compound is ring AB1. An exemplary scheme for preparing ring AB1 is as follows Figure 5 In one embodiment, refer to Figure 5 , ring A1 is activated, for example, by treatment with an activating agent such as trichloroacetonitrile or 2,2,2-trifluoro-N-phenyl-acetimidyl chloride, followed by addition of ring B1 and a Lewis acid such as bismuth trifluoroformate to give a glycosylated product from which the acetate protecting group is subsequently removed (using, for example, methanol in the presence of a carbonate base or using sodium methoxide). The resulting 1,2-diol functionality is then protected, for example, by conversion to a ketal. A suitable ketal is a ketal formed by reaction of a diol with cyclohexanone or cyclohexanone dimethyl ketal. The remaining B ring hydroxyl group may then be protected, for example, as a benzyl ether. The ketal protecting group may then be removed to give a 1,2-diol. In order to obtain a single free hydroxyl group of the desired ring AB1, the adjacent amino and hydroxyl groups are protected. A suitable protection strategy is to incorporate the amino and hydroxyl groups into an oxazolidinone. A suitable means of forming the oxazolidinone is to treat with a strong non-nucleophilic base such as sodium hydride to form an oxazolidinone and expel the benzyl alcohol.

[0407] In some embodiments, the AB ring compound is ring AB2. An exemplary scheme for preparing ring AB2 is as follows Figure 6 This scheme uses Figure 5 The same general strategy as provided in , but with ring A2 replacing ring A1.

[0408] In some embodiments, the AB ring compound is ring AB3. An exemplary scheme for preparing ring AB3 is as follows Figure 7 This scheme uses Figure 5 The same general strategy is provided in , but with ring A3 replacing ring A1.

[0409] Other AB ring compounds can be prepared in a similar manner, for example, using alternative A ring compounds (eg, Ring A4 or Ring A5), alternative B ring compounds, or both, and varying the protecting group strategy as desired.

[0410] Preparation of CD ring compounds

[0411] According to Scheme 1, an exemplary preparation of compounds of the present disclosure having an ABCD ring system (e.g., compounds of Formula (I) and Formula (III)) uses a CD ring compound. In some embodiments, the CD ring compound is ring CD1:

[0412]

[0413] Ring CD1 can be prepared according to the procedures reported in, for example, International Patent Application Publication No. WO2019 / 194858, assigned to Achaogen, Inc., which is incorporated herein by reference with respect to the preparation of CD ring compounds.

[0414] In some embodiments, the CD ring compound is ring CD2:

[0415]

[0416] Ring CD2 can be prepared according to the procedures reported in, for example, International Patent Application Publication No. WO2019 / 194858, assigned to Achaogen, Inc., which is incorporated herein by reference with respect to the preparation of CD ring compounds.

[0417] Preparation of ABCD ring compounds

[0418] The preparation of compounds having an ABCD ring system (i.e., compounds of formula (I) and formula (III)) according to Scheme 1 generally (and with reference to Scheme 1) comprises connecting the AB ring and the CD ring compounds via a glycosidic bond, each of which is as described above. As a non-limiting example, in a suitable solvent, the anomeric hydroxyl group of the CD ring compound is converted into a suitable leaving group (acceptor) that is capable of reacting with the hydroxyl group (donor) on the AB ring compound to form an interglycosidic bond and allowing the free hydroxyl group on the AB ring compound to react to form the bond. Examples of suitable leaving groups include, but are not limited to, halides, alkylthioates, arylthioates, imidates / salts, acetates / salts, phosphates / salts, O-pentenyls, mesylates / salts, tosylates / salts, and the like. In one embodiment, the CD ring compound is treated with an activator (e.g., trichloroacetonitrile) to form an imidate, and the AB ring compound is added, followed by an additional activator (e.g., Lewis acid). In some embodiments, the Lewis acid is trimethylsilyl triflate, trimethylaluminum, or boron trifluoride etherate.

[0419] In some embodiments, the AB ring compound is ring AB1, ring AB2, or ring AB3. In some embodiments, the CD ring compound is ring CD1. Exemplary schemes for preparing ring ABCD1, ring ABCD2, and ring ABCD3 are respectively Figure 8 , 9 and as shown in 10.

[0420] In one embodiment, the Figure 8 The scheme shown in is used to prepare ring ABCD1 (i.e., a precursor of the compound of formula (I)). Figure 8 , ring CD1 is activated by, for example, treatment with an activating agent such as trichloroacetonitrile, and then ring AB1 and a Lewis acid are added to obtain a compound having an ABCD ring system. In some embodiments, the Lewis acid is boron trifluoride etherate. After the glycosylation reaction, the protected amino and hydroxy groups are linked with an oxazolidinone and released by hydrolysis under alkaline conditions to obtain ring ABCD1.

[0421] In one embodiment, the Fig. 9 The scheme shown in is used to prepare ring ABCD2 (i.e., the precursor of the compound of formula (III)). Fig. 9 , ring CD1 is activated by, for example, treatment with an activating agent such as trichloroacetonitrile, and then ring AB2 and a Lewis acid are added to obtain a compound having an ABCD ring system. In some embodiments, the Lewis acid is boron trifluoride etherate. After the glycosylation reaction, the protected amino and hydroxy groups are linked with an oxazolidinone and released by hydrolysis under alkaline conditions to obtain ring ABCD2.

[0422] In one embodiment, the Fig.10 The scheme shown in is used to prepare ring ABCD3 (i.e., a precursor of the compound of formula (III)). This scheme utilizes the above-mentioned Fig. 9 The same transformation of , but replacing ring AB2 with ring AB3.

[0423] Other ABCD ring compounds can be prepared using a scheme similar to the above scheme, substituting the appropriate AB ring and CD ring compounds, while adjusting the protecting group strategy accordingly. Those skilled in the art will recognize the adjustment of the protecting group strategy to accommodate the various combinations of potential AB ring and CD ring compounds envisioned in the general formulas disclosed herein (e.g., (I) and (III)).

[0424] Introducing the substituent R b

[0425] In any of the foregoing embodiments, the ABCD compound (ie, the compound of formula (I) or formula (III)) carries a substituent R b In some embodiments, R b H, C 1-6 Alkyl, or having a structure according to formula (II).

[0426] In some embodiments, the ABCD compound is a compound of formula (III) with a substituent R b , where R b is H. In such an embodiment, the corresponding ABCD compound is subjected to comprehensive deprotection to obtain a compound of formula (III). In some embodiments, the comprehensive deprotection can be achieved, for example, by hydrogenolysis, thereby cleaving all benzyl protecting groups and reducing the azide to the corresponding amine. The remaining protecting groups (e.g., acetate) can be removed by acidic or basic hydrolysis before or after hydrogenolysis, or the remaining protecting groups can be simultaneously cleaved by methanolysis under hydrogenolysis conditions.

[0427] In some embodiments, the ABCD compound is a compound of formula (I) or formula (III), and has a substituent R b , where R b C 1-6Alkyl. In such an embodiment, the corresponding ABCD compound is subjected to reductive alkylation using the desired aldehyde and a suitable reducing agent (e.g., acetoxy or cyanoborohydride) to obtain the corresponding N-alkyl compound. The resulting N-alkyl compound is then subjected to comprehensive deprotection to obtain a compound of formula (I) or formula (III). In some embodiments, the comprehensive deprotection can be achieved by, for example, hydrogenolysis, thereby cutting off all benzyl protecting groups and reducing the azide to the corresponding amine. The remaining protecting groups (e.g., acetate) can be removed by acidic or alkaline hydrolysis before or after hydrogenolysis, or the remaining protecting groups can be cut off simultaneously by methanolysis under hydrogenolysis conditions.

[0428] In some embodiments, the ABCD compound is a compound of formula (I) or formula (III), and has a substituent R b , where R b Having a structure according to formula (II):

[0429]

[0430] in:

[0431] Q is NH, O or S;

[0432] n is an integer from 0 to 4;

[0433] R 1 is hydrogen or C 1-3 alkyl;

[0434] R 2 and R 3 Each occurrence is independently selected from: hydrogen, alkyl, halogen, and -OH; and R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b are each independently selected from hydrogen and C 1-3 Alkyl; or

[0435] R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

[0436] Therefore, in various embodiments, preparing the ABCD compound of formula (I) or formula (III) may include: b The reaction of a suitable protected precursor with the free NH group of the corresponding ABCD ring compound gives a substituted R c The ABCD ring compound, which after additional steps as shown, gives the desired substituent Rb An ABCD ring compound of formula (I) or formula (III).

[0437] Referring to the structure of formula (II), in some embodiments, Q is NH; R 1 is hydrogen; R 2 and R 3 Each occurrence is independently selected from hydrogen, alkyl, halogen, and -OH; R 4 is H. Therefore, in some embodiments, the substituent R c Having a structure according to formula (IIa):

[0438]

[0439] in:

[0440] Pg is a protecting group;

[0441] Z is an amine precursor or a protected amino group; and

[0442] The wavy line indicates the point of attachment to the amine of the B ring.

[0443] In some embodiments, Z is azide (N3). In some embodiments, Z is NHCBz.

[0444] In some embodiments, Pg is benzyl. In some embodiments, Pg is p-methoxybenzyl (PMB).

[0445] In the above embodiments, those skilled in the art will recognize that for the introduction of the substituent R c For example, a suitable reactant is a corresponding carboxylic acid or an activated form thereof (ie, the wavy line in the compound of formula (IIa) represents an OH group or a leaving group). Typically, the introduction of the substituent R c It is through the ABCD ring compound and R c In addition, the skilled person will recognize that the final comprehensive deprotection of the ABCD ring compound (e.g., by hydrogenolysis) will also remove all benzyl protecting groups and / or replace the substituent R c The azide is reduced to an amine to obtain the corresponding substituent R of the final ABCD ring compound of formula (I) or formula (III): b ,like Figure 11-13 shown.

[0446] In some embodiments, R b Having a structure selected from the following:

[0447]

[0448] Therefore, in some embodiments, the substituent R chas a structure according to formula (IIa), wherein n is 2; and R 2 and R 3 Each occurrence is independently selected from hydrogen and F. In such an embodiment, the substituent R c Having a structure selected from the following:

[0449]

[0450] In some embodiments, R b Selected from:

[0451] as well as

[0452] R c One of the following:

[0453]

[0454] In some embodiments, Rb has the following structure:

[0455]

[0456] The corresponding reactant is (S)-2-(benzyloxy)-4-((2-oxo-2-phenyl-1λ-ethyl)amino)butyric acid, which has the following structure:

[0457]

[0458] Both the reactant shown and the corresponding substituent Rb are referred to herein as "HABA" referring to the hydroxyaminobutyric acid portion of the compound.

[0459] In some embodiments, the substituent Rb has the following structure:

[0460]

[0461] The corresponding reactant is (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid, which has the following structure:

[0462]

[0463] Both the reactant shown and the corresponding substituent Rb are referred to herein as "F-HABA" referring to the fluoro-hydroxy-aminobutyric acid portion of the compound.

[0464] In one embodiment, (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid (F-HABA) can be prepared according to Fig.14 Prepared according to the protocol shown in . Fig.14, D-isoascorbic acid is protected as a ketal and cleaved oxidatively to give an α-hydroxy ester. The ester is then reduced to an alcohol and the primary hydroxyl group is converted to an azide. The secondary hydroxyl group is then converted to an F group with stereochemical inversion. The ketal is removed and the two hydroxyl groups are protected in different ways. For example, the primary hydroxyl group is protected as a silyl ether and the secondary hydroxyl group is protected as a benzyl ether (e.g., p-methoxybenzyl). The silyl ether is then cleaved and the primary hydroxymethyl group is oxidized to the corresponding carboxylic acid to give the desired (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid.

[0465] Pharmaceutical compositions and preparations

[0466] The compounds described herein may be considered useful pharmaceutical compositions for administration to a subject in need thereof. Pharmaceutical compositions may comprise at least a compound or salt as described herein, and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersants, suspending agents and / or thickening agents.

[0467] The pharmaceutical composition can be formulated using one or more physiologically acceptable carriers (including excipients or adjuvants). The formulation can be changed according to the selected route of administration. The pharmaceutical composition comprising the compound or salt as described herein can be manufactured by, for example, lyophilizing, mixing, dissolving, emulsifying, encapsulating or encapsulating the compound or salt. The pharmaceutical composition may also include the compound or salt described herein in free base form or in the form of a pharmaceutically acceptable salt.

[0468] The pharmaceutical compositions described herein may include at least one active ingredient (e.g., a compound or salt). The active ingredient may be encapsulated in microcapsules, which are prepared, for example, by coacervation techniques in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or macroemulsions, or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules).

[0469] The pharmaceutical compositions described herein may generally further comprise more than one active compound (e.g., compound or salt, and other agents) that is necessary for the particular indication being treated. The active compounds may have complementary activities that do not adversely affect each other. For example, the composition may also comprise a chemotherapeutic agent, a cytotoxic agent, a cytokine, a growth inhibitor, an antihormonal agent, an antiangiogenic agent, and / or a cardioprotectant. These molecules may be present in combination in an effective amount of the target effect.

[0470] The compositions and formulations may be sterilized. Sterilization may be accomplished by sterile filtration.

[0471] The compositions described herein can be formulated as injections for administration. Non-limiting examples of injection preparations may include sterile suspensions, solutions or emulsions in oily or aqueous vehicles. Suitable oily vehicles may include, but are not limited to, lipophilic solvents or carriers, such as fatty oils or synthetic fatty acid esters, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension. The suspension may also contain a suitable stabilizer. The injection may be formulated as a bolus injection or a continuous infusion. Alternatively, the compositions described herein may be lyophilized or in powder form to be reconstituted with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0472] For parenteral administration / application, the compound or salt can be combined with a pharmaceutically acceptable parenteral carrier and formulated into an injectable form of a unit dose (e.g., using a solution, suspension, emulsion). Such a carrier can be non-toxic and non-therapeutic in itself. The carrier can be water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous carriers such as fixed oils and ethyl oleate can also be used. Liposomes can be used as carriers. The carrier can contain a small amount of additives, such as substances that enhance isotonicity and chemical stability (e.g., buffers and preservatives).

[0473] Sustained-release preparations can also be prepared. Examples of sustained-release preparations may include semipermeable matrices of solid hydrophobic polymers, which may contain compounds or salts, and which may be in the form of shaped articles (e.g., films or microcapsules). Examples of sustained-release matrices may include: polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate), or poly(vinyl alcohol)), polylactides, copolymers of L-glutamic acid and γ-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers, such as LUPRON DEPOTM (i.e., injectable microspheres consisting of lactic acid-glycolic acid copolymers and leuprolide acetate) and poly-D-(-)-3-hydroxybutyric acid.

[0474] The pharmaceutical preparations described herein can be prepared by mixing the compound or salt with a pharmaceutically acceptable carrier, excipient and / or stabilizer for storage. The preparation may be a lyophilized preparation or an aqueous solution. At the dosage and concentration used, the acceptable carrier, excipient and / or stabilizer may be non-toxic to the recipient. Acceptable carriers, excipients and / or stabilizers may include buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; preservatives, polypeptides; proteins such as serum albumin or gelatin; hydrophilic polymers; amino acids; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes; and / or nonionic surfactants or polyethylene glycols.

[0475] In some embodiments, the drug delivery composition can be incorporated into a system comprising a substrate that delivers the composition to a site of administration, or a delivery site, or a treatment site. The substrate can be retained in the composition at the time of administration (or delivery of the composition) and for any time or thereafter indefinitely, or removed at the time of administration (or delivery of the composition) to leave the composition at the site of administration, or a delivery site, or a treatment site.

[0476] The compounds of formula (I) or (III) can be administered in "natural" form, or if desired, can be administered / administered in the form of salts, esters, amides, prodrugs, clathrates, derivatives, etc., provided that the salts, esters, amides, prodrugs, clathrates or derivatives are pharmacologically suitable, e.g., effective for treating pathologies and / or their various symptoms, e.g., as described herein. Salts, esters, amides, clathrates, prodrugs and other derivatives of compounds of formula (I) or (III) can be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry, as described, for example, by March (1992) Advanced Organic Chemistry - Reactions, Mechanisms and Structure, 4th Edition, Wiley-Interscience, New York, and as described above.

[0477] For example, pharmaceutically acceptable salts can be prepared for any compound of formula (I) or formula (III) described herein, which have a functional group capable of forming a salt. A pharmaceutically acceptable salt is any salt that retains the activity of the parent compound and does not produce any harmful or adverse effects on the subject to which it is administered and in the environment in which it is administered.

[0478] In various embodiments, pharmaceutically acceptable salts may be derived from organic or inorganic bases. The salts may be monovalent or polyvalent ions. Of particular interest are inorganic ions such as lithium, sodium, potassium, calcium, and magnesium. Organic salts may be made from amines, particularly ammonium salts such as monoalkylamines, dialkylamines, and trialkylamines, or ethanolamine. Salts may also be formed from caffeine, tromethamine, and similar molecules.

[0479] Methods for preparing the pharmaceutical compound of formula (I) or formula (III) into salts, esters, amides, prodrugs, etc. are well known to those skilled in the art. For example, a conventional method generally involving reaction with a suitable acid can be used to prepare a salt from a free base. Generally speaking, the base form of the drug is dissolved in a polar organic solvent (such as methanol or ethanol) and an acid is added thereto. The resulting salt or precipitate, or is taken out of the solution by adding a less polar solvent. The acid used to prepare the acid addition salt includes, but is not limited to, both organic acids and inorganic acids, such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc., and the inorganic acid is, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. The acid addition salt can be converted back into a free base by treating with a suitable base. Some particularly preferred acid addition salts of compounds of formula (I) or (III) herein include halide salts, such as salts that can be prepared using hydrochloric acid or hydrobromic acid. In contrast, formulations of basic salts of compounds of formula (I) or (III) disclosed herein are prepared in a similar manner using a pharmaceutically acceptable base, such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine, etc. Particularly preferred basic salts include alkali metal salts, such as sodium salts, and copper salts.

[0480] For preparing salt forms of basic drugs, the pKa of the counterion is preferably at least about 2 pH units lower than the pKa of the drug. Similarly, for preparing salt forms of acidic drugs, the pKa of the counterion is preferably at least about 2 pH units higher than the pKa of the drug. This allows the counterion to bring the pH of the solution to below pHmax (pH 最大 ) level in order to reach a salt plateau, where the solubility of the salt outweighs the solubility of the free acid or free base. The general rule of difference in pKa units of the ionizable groups in the active pharmaceutical ingredient (API) and the pKa units in the acid or base is intended to make proton transfer energetically favorable. When the pKa of the API and the counterion are not significantly different, solid complexes may form but may disproportionate (i.e., decompose into separate entities of drug and counterion) rapidly in an aqueous environment.

[0481] Preferably, the counterion is a pharmaceutically acceptable counterion. Suitable anionic salt forms include, but are not limited to, acetate, benzenesulfonate, benzoate, benzylate, bicarbonate, bitartrate, bitartrate, bromide, calcium edetate, camphorsulfonate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, propionate lauryl sulfate, ethanesulfonate, fumarate, glucoheptonate, gluconate, glutamate, glycolyl arsanoate, hexylresorcinol salt, halamine, hydrobromide, hydrochloride, hydroxy naphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, methanesulfonate, methylbromide, methylnitrate, methylsulfate, mucate, naphthenate, nitrate, pamoate (enbolate), pantothenate, phosphate and diphosphate, polygalacturonate, salicylate and disalicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, theoclorate, toluenesulfonate, triethyl iodide, valerate, and the like; and suitable cationic salt forms include, but are not limited to, aluminum, benzathine, calcium, ethylenediamine, lysine, magnesium, meglumine, potassium, procaine, sodium, tromethamine, zinc, and the like. Suitable cationic salt forms include, but are not limited to, benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and the like.

[0482] Preparation of esters generally involves functionalization of hydroxyl and / or carboxyl groups present in the molecular structure of the compound of formula (I) or formula (III). In certain embodiments, esters are generally acyl-substituted derivatives of free alcohol groups, i.e., moieties derived from carboxylic acids of formula RCOOH, wherein R is an alkyl group and preferably a lower alkyl group. If desired, the esters can be converted back into free acids using conventional hydrogenolysis or hydrolysis procedures.

[0483] Amides can also be prepared using techniques known to those skilled in the art or described in the relevant literature. For example, amides can be prepared from esters using suitable amine reactants, or they can be prepared from anhydrides or acid chlorides by reaction with ammonia or lower alkylamines.

[0484] In various embodiments, the compounds of Formula (I) or Formula (III) described herein can be used for parenteral administration / administration, topical administration / administration, oral administration / administration, nasal administration / administration (or inhalation in other ways), rectal administration / administration, or local administration / administration (e.g., by aerosol or transdermal administration / administration) for the prophylactic and / or therapeutic treatment of one or more pathologies / indications described herein (e.g., pathologies characterized by excessive amyloid plaque formation and / or deposition, or undesirable amyloid or pre-amyloid processing).

[0485] The compounds of formula (I) or formula (III) described herein may also be combined with a pharmaceutically acceptable carrier (excipient) to form a pharmacological composition. A pharmaceutically acceptable carrier may contain one or more physiologically acceptable compounds, which, for example, act to stabilize the composition or increase or decrease the absorption of the active agent. Physiologically acceptable compounds may include, for example: carbohydrates, such as glucose, sucrose or dextran, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, protective and uptake enhancers, such as lipids, compositions that reduce the clearance or hydrolysis of the active agent, or excipients, or other stabilizers and / or buffers.

[0486] Other physiologically acceptable compounds (especially those used in the preparation of tablets, capsules, gel caps, etc.) include, but are not limited to, binders, diluents / fillers, disintegrants, lubricants, suspending agents, and the like.

[0487] In certain embodiments, to make an oral dosage form (e.g., tablet), excipients (e.g., lactose, sucrose, starch, mannitol, etc.), optional disintegrants (e.g., calcium carbonate, calcium carboxymethylcellulose, sodium starch glycolate, crospovidone, etc.), binders (e.g., α-starch, gum arabic, microcrystalline cellulose, carboxymethylcellulose, polyvinyl pyrrolidone, hydroxypropyl cellulose, cyclodextrin, etc.), and optional lubricants (e.g., talc, magnesium stearate, polyethylene glycol 6000, etc.) are added to one or more active ingredients (e.g., compounds represented by the structure of Formula I, etc.), and the resulting composition is compressed. If necessary, the compressed product is coated (e.g., using known methods) to mask the taste or for enteric or sustained release. Suitable coating materials include, but are not limited to, ethyl cellulose, hydroxymethyl cellulose, Polyethylene glycol, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, and Eudragit (Rohm & Haas, Germany; methacrylic acid-acrylic acid copolymer).

[0488] Other physiologically acceptable compounds include wetting agents, emulsifiers, dispersants or preservatives, which are particularly useful for avoiding microbial growth or action. Various preservatives are well known, including, for example, phenol and ascorbic acid. It will be appreciated by those skilled in the art that the selection of pharmaceutically acceptable carriers (including physiologically acceptable compounds) depends on, for example, the route of administration of the activating agent and the specific physicochemical properties of the activating agent. In some embodiments, the excipient is sterile and generally free of undesirable substances. These compositions can be sterilized by conventional, well-known sterilization techniques. For various oral dosage form excipients (such as tablets and capsules), sterility is not required. USP / NF standards are generally sufficient.

[0489] According to the method of administration / application, the pharmaceutical composition can be administered / administered in a variety of unit dosage forms. Suitable unit dosage forms include, but are not limited to, powders, tablets, pills, capsules, lozenges, suppositories, patches, nasal sprays, injections, implantable sustained-release preparations, mucoadherent films, topical varnishes, lipid complexes, etc.

[0490] Pharmaceutical compositions comprising compounds of formula (I) or formula (III) can be manufactured by conventional mixing, dissolving, granulating, dragee making, levigating, emulsifying, encapsulating, embedding or lyophilizing processes. Pharmaceutical compositions can be prepared in a conventional manner using one or more physiologically acceptable carriers, diluents, excipients or adjuvants, which facilitate processing of the active agent into a pharmaceutically acceptable preparation. Suitable formulations depend on the selected route of administration.

[0491] In certain embodiments, the active agents described herein (e.g., compounds of formula (I) or formula (III)) and compositions comprising the compounds are formulated for oral administration / application. For oral administration / application, suitable formulations can be readily prepared by combining the active agent with a pharmaceutically acceptable carrier suitable for oral delivery as is well known in the art. Such carriers enable the active agents described herein to be formulated into tablets, pills, dragees, caplets, lozenges, capsules, liquids, gels, syrups, pastes, suspensions, etc., for oral ingestion by the patient to be treated. For oral solid formulations, such as powders, capsules, and tablets, suitable excipients may include fillers such as sugars (e.g., lactose, sucrose, mannitol, and sorbitol), cellulose preparations (e.g., corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose), synthetic polymers (e.g., polyvinyl pyrrolidone (PVP)), granulating agents; and binders. If desired, a disintegrant may be added, such as cross-linked polyvinyl pyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate. If desired, the solid dosage form may be coated with a sugar coating or an enteric coating using standard techniques. Preparation of enteric-coated particles is disclosed, for example, in U.S. Pat. Nos. 4,786,505 and 4,853,230.

[0492] For administration / application by inhalation, the active agent is conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer, using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules or cartridges (comprising, for example, 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.

[0493] In various embodiments, the active agent can be formulated into rectal or vaginal compositions, such as suppositories or retention enemas, containing, for example, 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 skilled in the art (see, for example, Allen (2007) Suppositories, Pharmaceutical Press), and generally involve mixing the active agent with a suitable base (e.g., hydrophilic substances (PEG), lipophilic materials such as cocoa butter or Witepsol W45, amphiphilic materials such as SUPPOCIRE AP and polyglycolized glycerides, etc.). The base is selected and compounded for the desired melting / delivery profile.

[0494] For topical administration / application, the compounds of formula (I) or formula (III) may be formulated into solutions, gels, ointments, creams, suspensions, etc. as is well known in the art.

[0495] In certain embodiments, the compounds of formula (I) or formula (III) described herein are formulated for systemic administration / application (e.g., as an injection) according to standard methods well known to those skilled in the art. Systemic formulations include, but are not limited to, those designed for administration by injection, such as subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, and those designed for transdermal, transmucosal oral, or pulmonary administration. For injection, the active agents described herein may be formulated in an aqueous solution, preferably in a physiologically compatible buffer (e.g., Hank's solution, Ringer's solution or saline buffer), and / or in certain emulsions. The solution may contain a formulation agent, such as a suspending agent, a stabilizer and / or a dispersant. In certain embodiments, the activator may be provided in powder form for use with a suitable carrier (e.g., sterile pyrogen-free water) prior to use. For transmucosal administration, and / or for blood / brain barrier passage, a penetrant suitable for passing through the barrier may be used in the formulation. Such penetrants are generally known in the art. Injectable and inhalable formulations are generally provided as sterile or substantially sterile formulations.

[0496] In addition to the above-described formulations, the active agent can also be formulated as a depot preparation. Such long-acting preparations can be administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. Thus, for example, the active agent can be formulated with a suitable polymer or hydrophobic material (e.g., as an emulsion in an acceptable oil) or an ion exchange resin, or formulated as a sparingly soluble derivative, for example, as a sparingly soluble salt.

[0497] In some embodiments, the activating agent described herein can also be delivered by skin using conventional transdermal drug delivery system, i.e. transdermal "patch", wherein the activating agent is generally contained in a laminated structure, which is used as a drug delivery device fixed on the skin. In this structure, the pharmaceutical composition is generally contained in a layer or "reservoir" below the upper backing layer. It will be understood that in this context, the term "reservoir" refers to a certain amount of "active ingredient" that can eventually be used for delivery to the skin surface. Therefore, for example, the "reservoir" may include the active ingredient in the adhesive on the backing layer of the patch, or in any of the various different matrix preparations known to persons skilled in the art. The patch may contain a single reservoir, or may contain multiple reservoirs.

[0498] In an illustrative embodiment, the storage body comprises a polymer matrix, and this polymer matrix is ​​a pharmaceutically acceptable contact adhesive material, and this polymer matrix is ​​used for fixing the system to the skin during drug delivery.The example of suitable skin contact adhesive material includes but is not limited to polyethylene, polysiloxane, polyisobutylene, polyacrylate, polyurethane etc.Alternatively, the storage body containing medicine and skin contact adhesive exist as separate and different layers, wherein adhesive is below the storage body, and in this case, the storage body can be a polymer matrix as described above, or can be a liquid or hydrogel storage body, or take some other forms.The backing layer in these laminates serves as the upper surface of device, is preferably used as the main structural element of " patch ", and provides most flexibility for this device.The material selected for the backing layer is preferably substantially impermeable to the activating agent and any other material existing.

[0499] Alternatively, other drug delivery systems may be used. For example, liposomes, emulsions, and microemulsions / nanoemulsions are well-known examples of delivery vehicles that can be used to protect and deliver pharmaceutically active compounds. Certain organic solvents, such as dimethyl sulfoxide (DMSO), may also be used, although usually at the expense of greater toxicity.

[0500] In certain embodiments, the compounds of Formula (I) or Formula (III) as described herein are formulated into nanoemulsions. Nanoemulsions include, but are not limited to, oil-in-water (O / W) nanoemulsions and water-in-oil (W / O) nanoemulsions. A nanoemulsion can be defined as an emulsion having an average droplet diameter ranging from about 20 to about 1000 nm. Typically, the average droplet size is between about 20 nm and about 500 nm or between about 50 nm and about 500 nm. The terms submicron emulsion (SME) and mini-emulsion are used as synonyms.

[0501] Illustrative oil-in-water (O / W) nanoemulsions include, but are not limited to: surfactant micelles - micelles composed of small molecule surfactants or detergents (e.g., SDS / PBS / 2-propanol); polymer micelles - micelles composed of polymer, copolymer or block copolymer surfactants (e.g., Pluronic L64 / PBS / 2-propanol); mixed micelles - micelles in which more than one surfactant component is present, or in which one of the liquid phases (usually an alcohol or fatty acid compound) participates in the formation of micelles (e.g., octanoic acid / PBS / EtOH); integral micelles - mixed micelles in which the active agent serves as a co-surfactant, forming an integral part of the micelle; and Pickering (solid phase) emulsions / emulsions - emulsions in which the active agent is associated with the exterior of solid nanoparticles (e.g., polystyrene nanoparticles / PBS / oil-free phase).

[0502] Illustrative water-in-oil (W / O) nanoemulsions include, but are not limited to: surfactant micelles - micelles composed of small molecule surfactants or detergents (e.g., dioctyl sulfosuccinate / PBS / 2-propanol, isopropyl myristate / PBS / 2-propanol, etc.); polymer micelles - micelles composed of polymer, copolymer or block copolymer surfactants (e.g., L121 / PBS / 2-propanol); mixed micelles - micelles in which more than one surfactant component is present, or one of the liquid phases (usually an alcohol or fatty acid compound) participates in the formation of the micelle (e.g., capric / caprylic diglyceride / PBS / EtOH); integrated micelles - mixed micelles in which the active agent acts as a co-surfactant, forming an integral part of the micelle (e.g., active agent / PBS / polypropylene glycol); and Pickering (solid phase) emulsions / emulsions - emulsions in which the active agent is externally associated with solid nanoparticles (e.g., chitosan nanoparticles / anhydrous phase / mineral oil).

[0503] As described above, in certain embodiments, the nanoemulsion comprises one or more surfactants or detergents. In some embodiments, the surfactant is a non-anionic detergent (e.g., a polysorbate surfactant, a polyoxyethylene ether, etc.). The surfactant used in the present invention includes, but is not limited to, surfactants such as the following: and Family of compounds.

[0504] In certain embodiments, the emulsion further comprises one or more cationic halogen-containing compounds, including but not limited to cetylpyridinium chloride. In further embodiments, the composition further comprises one or more compounds that increase the interaction of the composition with the microorganism (e.g., chelating agents such as ethylenebis(oxyethylenenitrilo)tetraacetic acid, or ethylenediaminetetraacetic acid in a buffer) ("interaction enhancers").

[0505] In some embodiments, the nanoemulsion further comprises an emulsifier to help form the emulsion. Emulsifiers include certain compounds that aggregate at the oil / water interface to form a continuous film to prevent direct contact between two adjacent droplets. Certain embodiments of the present invention are characterized by an oil-in-water emulsion composition that can be easily diluted with water to a desired concentration without compromising its anti-pathogenic properties.

[0506] In addition to discrete oil droplets dispersed in an aqueous phase, certain oil-in-water emulsions may also contain other lipid structures, such as small lipid vesicles (e.g., lipid globules that are typically composed of several substantially concentric lipid bilayers separated from each other by layers of aqueous phase), micelles (e.g., amphiphilic molecules in small clusters of 50-200 molecules arranged with the polar head groups facing toward the aqueous phase and the nonpolar tails sequestered inwardly away from the aqueous phase), or lamellar phases (lipid dispersions in which each particle is composed of parallel amphiphilic bilayers separated by a thin film of water).

[0507] These lipid structures are formed due to the hydrophobic forces that drive non-polar residues (e.g., long hydrocarbon chains) from water. The above lipid formulations can be generally described as surfactant lipid formulations (SLPs). SLPs are minimally toxic to mucosa, and are believed to be metabolized in the small intestine (see, e.g., Hamouda et al., (1998) J. Infect. Disease 180:1939).

[0508] In certain embodiments, the emulsion comprises a discontinuous oil phase distributed in an aqueous phase, a first component comprising an alcohol and / or glycerol, and a second component comprising a surfactant or a halogen-containing compound. The aqueous phase may comprise any type of aqueous phase, including but not limited to: water (e.g., deionized water, distilled water, tap water) and a solution (e.g., a phosphate buffered saline solution, or other buffer system). The oil phase may comprise any type of oil, including but not limited to vegetable oils (e.g., soybean oil, avocado oil, linseed oil, coconut oil, cottonseed oil, squalene oil, olive oil, rapeseed oil, corn oil, rapeseed oil, safflower oil, and sunflower oil), animal oils (e.g., fish oil), flavor oils, water-insoluble vitamins, mineral oils, and motor oils. In certain embodiments, the oil phase accounts for 30-90% by volume of the oil-in-water emulsion (i.e., 30-90% of the total volume of the final emulsion), more preferably 50-80%. The formulation need not be limited to a specific surfactant, but in certain embodiments, the surfactant is: a polysorbate surfactant (e.g., and ), phenoxypolyethoxyethanol (e.g., X-100, X-301, X-165, X-102 and X-200 and ), or sodium lauryl sulfate, etc.

[0509] In certain embodiments, there is a halogen-containing component. The nature of the halogen-containing compound, in some preferred embodiments, the halogen-containing compound includes: chloride salts (e.g., NaCl, KCl, etc.), hexadecylpyridinium halides, hexadecyltrimethylammonium halides, hexadecyldimethylethylammonium halides, hexadecyldimethylbenzylammonium halides, hexadecyltributylphosphonium halides, dodecyltrimethylammonium halides, tetradecyltrimethylammonium halides, hexadecylpyridinium chloride, hexadecyltrimethylammonium chloride, hexadecylbenzylammonium chloride, hexadecylpyridinium bromide, hexadecyltrimethylammonium bromide, hexadecyldimethylethylammonium bromide, hexadecyltributylphosphonium bromide, dodecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide, and the like.

[0510] In certain embodiments, the emulsion comprises a quaternary ammonium compound. Quaternary ammonium compounds include, but are not limited to: N-alkyl dimethyl benzyl ammonium saccharinate, 1,3,5-triazine-1,3,5 (2H, 4H, 6H) -triethanol; 1-decyl betaine (decanaminium), N-decyl-N, N-dimethyl-, chloride (or) didecyl dimethyl ammonium chloride; 2-(2-(p-(diisobutyl) tolyloxy (cresosxy)) ethoxy) ethyl dimethyl benzyl ammonium chloride; 2-(2-(p-(diisobutyl) phenoxy) ethoxy) ethyl dimethyl benzyl ammonium chloride; alkyl 1 or 3 benzyl-1-(2-hydroxyethyl)-2-imidazolium chloride; alkyl bis (2-hydroxyethyl) benzyl ammonium chloride; alkyl dimethyl benzyl ammonium chloride; Alkyl dimethyl 3,4-dichlorobenzylammonium chloride (100% C12); Alkyl dimethyl 3,4-dichlorobenzylammonium chloride (50% C14, 40% C12, 10% C16); Alkyl dimethyl 3,4-dichlorobenzylammonium chloride (55% C14, 23% C12, 20% C16); Alkyl dimethyl benzyl ammonium chloride; Alkyl dimethyl benzyl ammonium chloride (100% C14); Alkyl dimethyl benzyl ammonium chloride (100% C16); Alkyl dimethyl benzyl ammonium chloride (41% C14, 28% C12); Alkyl dimethyl benzyl ammonium chloride (47% C12, 18% C14); Alkyl dimethyl benzyl ammonium chloride (55% C16, 20% C14); Alkyl Dimethylbenzylammonium chloride (58% C14, 28% C16); Alkyldimethylbenzylammonium chloride (60% C14, 25% C12); Alkyldimethylbenzylammonium chloride (61% C11, 23% C14); Alkyldimethylbenzylammonium chloride (61% C12, 23% C14); Alkyldimethylbenzylammonium chloride (65% C12, 25% C14); Alkyldimethylbenzylammonium chloride (67% C12, 24% C14); Alkyldimethylbenzylammonium chloride (67% C12, 25% C14); Alkyldimethylbenzylammonium chloride (90% C14, 5% C12); Alkyldimethylbenzylammonium chloride (93% C14, 4% C12); Alkyldimethyl Benzyl ammonium chloride (95% C16, 5% C18); alkyl dimethyl benzyl ammonium chloride (and) didecyl dimethyl ammonium chloride; alkyl dimethyl benzyl ammonium chloride (as in fatty acids); alkyl dimethyl benzyl ammonium chloride (C12-C16); alkyl dimethyl benzyl ammonium chloride (C12-C18); alkyl dimethyl benzyl and dialkyl dimethyl ammonium chlorides; alkyl dimethyl dimethyl benzyl ammonium chloride; alkyl dimethyl ethyl ammonium bromide (90% C14, 5% C16, 5% C12); alkyl dimethyl ethyl ammonium bromide (mixed alkyl and alkenyl, as in the fatty acids of soybean oil); alkyl dimethyl ethyl benzyl ammonium chloride; alkyl dimethyl ethyl benzyl ammonium chloride (60% C14);Alkyl dimethyl isopropyl benzyl ammonium chloride (50% C12, 30% C14, 17% C16, 3% C18); Alkyl trimethyl ammonium chloride (58% C18, 40% C16, 1% C14, 1% C12); Alkyl trimethyl ammonium chloride (90% C18, 10% C16); Alkyl dimethyl (ethyl benzyl) ammonium chloride (C12-18); Di-(C8-10)-alkyl dimethyl ammonium chloride; Dialkyl dimethyl ammonium chloride; Dialkyl dimethyl ammonium chloride; Dialkyl dimethyl ammonium chloride; Dialkyl dimethyl ammonium chloride; Dialkylmethylbenzylammonium chloride; didecyldimethylammonium chloride; diisodecyldimethylammonium chloride; dioctyldimethylammonium chloride; dodecylbis(2-hydroxyethyl)octylammonium chloride; dodecyldimethylbenzylammonium chloride; dodecylcarbamoylmethyldimethylbenzylammonium chloride; heptadecylhydroxyethylimidazolium chloride; hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine; myristalkonium chloride (and) Quaternium 14; N,N-dimethyl-2-hydroxypropylammonium chloride polymer; n-alkyldimethylbenzylammonium chloride; n-alkyldimethylethylbenzylammonium chloride; n-tetradecyldimethylbenzylammonium chloride monohydrate; octyldecyldimethylammonium chloride; octyldodecyldimethylammonium chloride; octylphenoxyethoxyethyldimethylbenzylammonium chloride; oxydiethylenebis(alkyldimethylammonium chloride); quaternary ammonium compounds, dicocoylalkyldimethyl, chloride; trimethoxysilylpropyldimethyloctadecylammonium chloride; trimethoxysilylquaternary ammonium salt, trimethyldodecylbenzylammonium chloride; n-dodecyldimethylethylbenzylammonium chloride; n-hexadecyldimethylbenzylammonium chloride; n-tetradecyldimethylbenzylammonium chloride; n-tetradecyldimethylethylbenzylammonium chloride; and n-octadecyldimethylbenzylammonium chloride. ;

[0511] Nanoemulsion formulations and methods of making them are well known to those skilled in the art and are described, for example, in U.S. Pat. Nos. 7,476,393, 7,468,402, 7,314,624, 6,998,426, 6,902,737, 6,689,371, 6,541,018, 6,464,990, 6,461,625, 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, and 5,152,923, and Fanun et al. (2009) Microemulsions: Properties and Applications (Surfactant Science), CRC Press, Boca Raton, FL.

[0512] In certain embodiments, one or more of the active agents described herein may be provided as a "concentrate," e.g., in a storage container (e.g., in a pre-measured volume) ready for dilution, or in a soluble capsule ready for addition of a volume of water, alcohol, hydrogen peroxide, or other diluent.

[0513] In certain embodiments, the compounds of formula (I) or formula (III) as described herein are formulated as inclusion complexes. Although not limited to cyclodextrin inclusion complexes, it is noted that cyclodextrin is the most commonly used agent for forming drug inclusion complexes. Cyclodextrin (CD) is a cyclic oligomer of glucose, typically containing 6, 7, or 8 glucose monomers, which are linked by α-1,4 bonds. These oligomers are generally referred to as α-CD, β-CD, and γ-CD, respectively. Higher oligomers containing up to 12 glucose monomers are known and are contemplated in the formulations described herein. Functionalized cyclodextrin inclusion complexes are also contemplated. Illustrative but non-limiting functionalized cyclodextrins include, but are not limited to: sulfonates, sulfonates and sulfinates, or disulfonates of hydroxybutenyl cyclodextrins; sulfonates, sulfonates and sulfinates, or disulfonates of mixed ethers of cyclodextrins, wherein at least one ether substituent is hydroxybutenyl cyclodextrin. Exemplary cyclodextrins include: polysaccharide ethers, which include at least one 2-hydroxybutenyl substituent, wherein at least one hydroxybutenyl substituent is sulfonated and sulfinated, or disulfonated, and alkyl polyglycoside ethers, which include at least one 2-hydroxybutenyl substituent, wherein at least one hydroxybutenyl substituent is sulfonated and sulfinated, or disulfonated. In various embodiments, inclusion compounds formed between sulfonated hydroxybutenyl cyclodextrin and one or more activating agents described herein are contemplated. Methods for preparing cyclodextrins and cyclodextrin inclusion compounds are found in, for example, U.S. Patent Publication No. 2004 / 0054164 and documents cited therein, and U.S. Patent Publication No. 2011 / 0218173 and documents cited therein.

[0514] In certain embodiments, the compounds of Formula (I) or Formula (III) described herein may also be administered using medical devices known in the art. For example, in one embodiment, the pharmaceutical compositions of the present invention may be administered using a needle-free subcutaneous injection device, such as those disclosed in: U.S. Patent No. 5,399,163; U.S. Patent No. 5,383,851; U.S. Patent No. 5,312,335; U.S. Patent No. 5,064,413; U.S. Patent No. 4,941,880; U.S. Patent No. 4,790,824; or U.S. Patent No. 4,596,556. Well-known examples of implants and modules that can be used for such delivery include, but are not limited to: U.S. Patent No. 4,487,603, which discloses an implantable microinfusion pump for dispensing drugs at a controlled rate; U.S. Patent No. 4,486,194, which discloses a therapeutic device for administering drugs through the skin; U.S. Patent No. 4,447,233, which discloses a drug infusion pump for delivering drugs at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow implantable infusion device for continuous drug delivery; U.S. Patent No. 4,439,196, which discloses an osmotic drug delivery system with multiple chamber compartments; and U.S. Patent No. 4,475,196, which discloses an osmotic drug delivery system. In a specific embodiment, a compound of formula (I) or formula (III) can be administered using a drug-eluting stent, such as those described in WO 01 / 87263 and related disclosures or those described by Perin (Perin, EC, 2005). Many other such implants, delivery systems and modules are known to those skilled in the art.

[0515] As described herein, the dosage of the compound of formula (I) or formula (III) will vary depending on the specific compound, the disease involved, the subject, the nature and severity of the disease, and the physical condition of the subject, and the selected dosage / administration route. Appropriate dosage can be easily determined by those skilled in the art.

[0516] In certain embodiments, the composition may contain from 0.1%, such as 0.1-70%, or 5-60%, or 10-30%, of one or more compounds of Formula (I) or Formula (III), depending on the method of administration.

[0517] Those skilled in the art will recognize that the optimal amount and interval of a compound of formula (I) or formula (III) as described herein will depend on the nature and extent of the condition being treated, the form, route and site of administration, and the age and condition of the specific subject being treated, and that the physician will ultimately determine the appropriate dose to be used. The dose may be repeated as appropriate. If side effects occur, the amount and / or frequency of the dose may be altered or reduced in accordance with normal clinical practice.

[0518] In some embodiments, pharmaceutically acceptable salts may also include both acid addition salts and base addition salts. In some embodiments, "pharmaceutically acceptable acid addition salts" may refer to salts that retain the biological effectiveness and properties of free bases, are not biologically or otherwise undesirable, and can be formed from inorganic acids and organic acids, such as but not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; organic acids such as but not limited to acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid. Acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, etc.

[0519] In some embodiments, "pharmaceutically acceptable base addition salts" may refer to those salts that retain the biological effectiveness and properties of the free acid, which are not biologically or otherwise undesirable. These salts may be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases may include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. For example, inorganic salts may include, but are not limited to, ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases may include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, arginine, histidine, caffeine, procaine, hydrazine, choline, betaine, phenylethylamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like.

[0520] For the purpose of administration, the compounds of the present disclosure can be applied as raw chemicals, or can be formulated into pharmaceutical compositions. The pharmaceutical compositions of the present disclosure include one or more compounds of formula (I) or formula (III), or any pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, and pharmaceutically acceptable carriers. The one or more compounds of formula (I) or formula (III), or any pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, are present in an amount effective for treating a specific disease or target condition-for example, in an amount sufficient to treat a bacterial infection, and toxicity is generally acceptable to the subject. In some embodiments, the one or more compounds of formula (I) or formula (III), or any pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, are present in the composition in an amount effective for treating a specific disease or target condition. The antibacterial activity of a compound of formula (I) or formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof can be determined by a person skilled in the art, for example, as described in the Examples herein. Appropriate concentrations and dosages can be easily determined by a person skilled in the art.

[0521] Administration of one or more compounds of the present disclosure (e.g., compounds of Formula (I) or Formula (III)) or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers of any of them in pure form or in an appropriate pharmaceutical composition can be carried out by any accepted mode of administration that exerts a similar effect. In some embodiments, the one or more compounds of the present disclosure administered are compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers of any of the foregoing, administered in pure form or in an appropriate pharmaceutical composition.

[0522] The pharmaceutical composition of the present disclosure can be prepared as follows: one or more compounds of the present disclosure, or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof, are combined with an appropriate pharmaceutically acceptable carrier, and can be formulated into a preparation in solid, semisolid, liquid or gaseous form, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres and aerosols. Typical routes of administration of such pharmaceutical compositions may include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, oral, rectal, vaginal, and intranasal. The term "parenteral" as used herein may include subcutaneous injection, intravenous, intramuscular, intrasternal injection or infusion techniques. The pharmaceutical composition of the present disclosure can be formulated so as to allow the active ingredients contained therein to be bioavailable when the composition is administered to a subject. The composition to be administered to the subject or patient takes the form of one or more dosage units, where, for example, a tablet can be a single dosage unit, while a container containing one or more compounds of the present disclosure or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof in aerosol form can contain multiple dosage units. Actual methods of preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000).

[0523] The pharmaceutical compositions of the present disclosure may be in solid or liquid form. In one aspect, the carrier is a particulate matter, such that the composition is in the form of, for example, a tablet or powder. The carrier may be a liquid, wherein the composition is, for example, an oral syrup, an injectable liquid, or an aerosol, which may be used, for example, for inhalation administration.

[0524] When intended for oral administration, the pharmaceutical compositions of the present disclosure are typically in solid or liquid form, wherein the forms considered herein as solid or liquid may include semi-solid, semi-liquid, suspension, and gel forms.

[0525] As a solid composition for oral administration, the pharmaceutical composition can be formulated into powder, granules, compressed tablets, pills, capsules, chewing gum, wafers or similar forms. Such solid compositions will generally contain one or more inert diluents or edible carriers. In addition, one or more of the following may be present: a binder, such as carboxymethyl cellulose, ethyl cellulose, microcrystalline cellulose, tragacanth gum or gelatin; an excipient, such as starch, lactose or dextrin, a disintegrant, such as alginic acid, sodium alginate, Corn starch, etc.; Lubricants, such as magnesium stearate or Glidants, such as colloidal silicon dioxide; sweeteners, such as sucrose or saccharin; flavoring agents, such as mint, methyl salicylate, or orange flavoring; and coloring agents.

[0526] If the pharmaceutical composition is in the form of a capsule (eg, a gelatin capsule), the pharmaceutical composition may contain, in addition to the materials disclosed herein, a liquid carrier such as polyethylene glycol or oil.

[0527] Pharmaceutical compositions of the present disclosure may be in liquid form, such as elixirs, syrups, solutions, emulsions or suspensions. As two examples, the liquid may be used for oral administration or for delivery by injection. If intended for oral administration, pharmaceutical compositions of the present disclosure may also generally contain one or more of sweeteners, preservatives, dyes / colorants and flavor enhancers in addition to one or more compounds of the present disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers or their tautomers. Intentions may include one or more of surfactants, preservatives, wetting agents, dispersants, suspending agents, buffers, stabilizers and isotonic agents in compositions for administration by injection.

[0528] Liquid pharmaceutical compositions of the present disclosure, whether solutions, suspensions or other similar forms, may include one or more of the following adjuvants: sterile diluents, such as water for injection, saline solution, physiological saline, Ringer's solution, isotonic sodium chloride, non-volatile oils, such as synthetic monoglycerides or diglycerides that can serve as solvents or suspension media, polyethylene glycol, glycerol, propylene glycol or other solvents; antibacterial agents, such as benzyl alcohol or parabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates or phosphates, and agents for adjusting tension, such as sodium chloride or dextrose. Parenteral formulations may be packaged in ampoules, disposable syringes or multi-dose vials made of glass or plastic. In some embodiments, the adjuvant is physiological saline. In some embodiments, the injectable pharmaceutical composition is sterile.

[0529] Liquid pharmaceutical compositions of the present disclosure for parenteral or oral administration should contain an amount of one or more compounds of the present disclosure, or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof, so that a suitable dosage is obtained.

[0530] Pharmaceutical compositions of the present disclosure may be intended for topical administration, in which case the carrier may suitably include a solution, emulsion, ointment or gel substrate. For example, the substrate may include one or more of the following substances: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents (such as water and alcohol), and emulsifiers and stabilizers. Thickeners may be present in the pharmaceutical compositions for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or an iontophoresis device.

[0531] Pharmaceutical compositions of the present disclosure may be intended for rectal administration, in the form of, for example, suppositories, which will melt and release the drug in the rectum. Compositions for rectal administration may include an oleaginous base as a suitable non-irritating excipient. Such bases may include, but are not limited to, lanolin, cocoa butter, and polyethylene glycol.

[0532] Pharmaceutical compositions of the present disclosure may include various materials that change the physical form of solid or liquid dosage units. For example, the composition may include a material that forms a coating shell around the active ingredient. The material that forms the coating shell is usually inert and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be encapsulated in a gelatin capsule.

[0533] Solid or liquid forms of pharmaceutical compositions of the present disclosure may include agents that bind to one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof to assist in delivering the one or more compounds or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof. Suitable agents that may play this role may include monoclonal or polyclonal antibodies, proteins, or liposomes.

[0534] The pharmaceutical composition of the present disclosure can be prepared into a dosage unit that can be used as an aerosol. The term "aerosol" is used to represent various systems ranging from systems with colloidal properties to systems consisting of pressurized packaging. Delivery can be carried out by liquefied or compressed gas or by a suitable pump system that distributes the active ingredient. Aerosols of one or more compounds of the present disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers or tautomers can be delivered in a single-phase, two-phase or three-phase system to deliver the active ingredient. The delivery of an aerosol may include: necessary containers, activators, valves, subcontainers, etc., which together form a test kit. Those skilled in the art can determine the preferred aerosol without excessive experimentation.

[0535] The pharmaceutical composition of the present disclosure can be prepared by methods well known in the pharmaceutical field. For example, a pharmaceutical composition intended to be administered by injection can be prepared by mixing one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof with sterile distilled water to form a solution. A surfactant can be added to promote the formation of a uniform solution or suspension. A surfactant is a compound that non-covalently interacts with one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof to promote the dissolution or uniform suspension of the one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof in an aqueous delivery system.

[0536] In some embodiments, the pharmaceutical composition of the present disclosure can be enriched to provide mainly one enantiomer of the compound described herein. The mixture enriched with enantiomers can contain, for example, at least 60 mol % of one enantiomer, or at least 75 mol %, at least 80 mol %, at least 85 mol %, at least 90 mol %, at least 95 mol %, at least 96 mol %, at least 97 mol %, at least 98 mol %, at least 99 mol %, at least 99.5 mol %, or even 100 mol % of one enantiomer. In certain embodiments, the compound described herein enriched in one enantiomer can be substantially free of another enantiomer, wherein substantially free of can mean that the substance in question accounts for less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% compared to the amount of other enantiomers 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 can be said to contain 98 mole percent of the first enantiomer and only 2 percent of the second enantiomer.

[0537] In some embodiments, the pharmaceutical composition of the present disclosure can be enriched to provide mainly one diastereomer of the compound disclosed herein. A diastereomer-enriched mixture can contain, for example, at least 60 mol % of one diastereomer, or at least 75 mol %, at least 80 mol %, at least 85 mol %, at least 90 mol %, at least 95 mol %, at least 96 mol %, at least 97 mol %, at least 98 mol %, at least 99 mol %, at least 99.5 mol %, or even 100 mol % of one diastereomer. In certain embodiments, the compound described herein that is enriched in one diastereomer can be substantially free of other diastereomers, wherein "substantially free" can mean that the substance in question accounts for less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% compared to the amount of other diastereomers in, for example, a pharmaceutical composition or a mixture of compounds.

[0538] Diastereomeric mixtures can be separated into their respective diastereomers according to their physicochemical differences by methods well known to those skilled in the art, such as by chromatography and / or fractional crystallization. Enantiomers can be separated by reacting with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acyl chloride) to convert the enantiomeric mixture into a diastereomeric mixture, separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers into the corresponding pure enantiomers. Enantiomers can also be separated by using a chiral HPLC column. In addition, some compounds of the present disclosure may be atropisomers or rotameric forms and are considered to be part of the present disclosure.

[0539] Therapeutic applications

[0540] The compounds, compositions and methods of the present disclosure can be used in a variety of different subjects, including, but not limited to, mammals, humans, non-human mammals, domesticated animals (e.g., laboratory animals, household pets, or livestock), non-domesticated animals (e.g., wild animals), dogs, cats, rodents, mice, hamsters, cows, birds, chickens, fish, pigs, horses, goats, sheep, rabbits, or any combination thereof.

[0541] The compounds and compositions described herein can be used as therapeutic agents, e.g., therapeutic agents that can be administered to a subject in need thereof. The therapeutic effects of the present disclosure can be obtained in a subject by reducing, inhibiting, alleviating, or eradicating a disease state (including, but not limited to, its symptoms). The therapeutic effect on a subject suffering from a disease or condition, or susceptible to or beginning to suffer from the disease or condition, can be obtained by reducing, inhibiting, preventing, alleviating, or eradicating the disease or condition, or a pre-disease condition or pre-disease state.

[0542] In practicing the methods described herein, a therapeutically effective amount of the compositions described herein can be administered to a subject in need thereof, typically for the treatment and / or prevention of a condition or progression thereof. The pharmaceutical composition can affect the subject's physiology, such as the immune system, inflammatory response, or other physiological effects. The therapeutically effective amount can vary depending on the severity of the disease, the age and relative health of the subject, the efficacy of the compound used, and other factors.

[0543] Treatment and / or treatment (in) may refer to any successful signs of treating or ameliorating a disease or condition. Treatment may include, for example, reducing, delaying or alleviating the severity of one or more symptoms of a disease or condition, or it may include reducing the frequency with which a patient experiences symptoms of a disease, defect, condition or adverse condition. Herein, treatment may be used to refer to a method that results in treating or ameliorating a disease or condition at some level, and a range of results for this purpose may be considered, including but not limited to preventing the condition altogether.

[0544] Prevent (prevent, preventing, etc.) can refer to preventing a disease or condition, such as tumor formation, in a patient. For example, if an individual at risk for developing a tumor or other form of cancer is treated with the methods of the present disclosure and does not subsequently develop a tumor or other form of cancer, the disease has been prevented in the individual, at least for a period of time. Preventing can also refer to preventing a patient who has been previously treated for a disease or condition from developing the disease or condition again (e.g., by preventing recurrence).

[0545] A therapeutically effective amount may be an amount of the composition or its active ingredient sufficient to provide a beneficial effect or otherwise reduce harmful non-beneficial events to an individual receiving the composition. A therapeutically effective dose may be a dose that produces one or more desired or required (e.g., beneficial) effects, such administration occurring once or multiple times within a given time. The exact dose may depend on the purpose of the treatment and may be determined by those skilled in the art using known techniques.

[0546] Treatments for bacterial infections

[0547] The disclosure features methods and uses for treating bacterial infections comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of the disclosure, or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof, or administering a therapeutically effective amount of a pharmaceutical composition comprising one or more compounds of the disclosure, or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof.

[0548] The compounds of the present disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers or tautomers, and the pharmaceutical compositions of the present disclosure (e.g., compounds of formula (I) or formula (III)) may possess antibacterial activity against a broad 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 may be inhibited by the compounds of the present disclosure, including, but not limited to, Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacter, Neisseria, Bacillus, Peptococcus, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Francisella, Yersinia, Corynebacterium, Moraxella, Enterococcus, Burkholderia, and other organisms.

[0549] Therefore, the compounds disclosed herein, or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, and pharmaceutical compositions comprising the compounds or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, can be used, for example, to treat bacterial infections and to treat conditions caused by the bacterial infections. According to the present disclosure, bacterial infections susceptible to treatment may include primary infections and mixed infections caused by one bacterium and one or more additional infectious agents (e.g., bacteria, viruses, parasites and fungi).

[0550] In one aspect, a method for treating a bacterial infection is provided, the method comprising administering to a subject in need thereof a therapeutically effective amount of one or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof, or a pharmaceutical composition comprising one or more compounds of Formula (I) or Formula (III) or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer thereof.

[0551] In some embodiments, the bacterial infection is an infection caused by a species of Staphylococcus, Lactobacillus, Streptococcus, Sarcina, Escherichia, Enterobacter, Klebsiella, Pseudomonas, Acinetobacter, Mycobacterium, Proteus, Campylobacter, Citrobacil, or other similar species. cter species, Neisseria species, Bacillus species, Peptococcus species, Salmonella species, Shigella species, Serratia species, Haemophilus species, Brucella species, Francisella species, Yersinia species, Corynebacterium species, Moraxella species, Burkholderia species, or Enterococcus species.

[0552] In some embodiments, the bacterial infection is caused by Gram-positive bacteria, Gram-negative bacteria, aerobic bacteria, or facultative anaerobic bacteria.

[0553] In some embodiments, the bacterial infection is caused by gram-positive bacteria. Examples of the gram-positive bacteria include, but are not limited to, Staphylococcus species, Lactobacillus species, Streptococcus species, Sarcina species, Bacillus species, Peptococcus species, Corynebacterium species, and Enterococcus species. In some embodiments, the gram-positive bacterial infection is an infection caused by Staphylococcus species, Lactobacillus species, Streptococcus species, Sarcina species, Bacillus species, Peptococcus species, Corynebacterium species, or Enterococcus species. In some embodiments, the bacterial infection is caused by Staphylococcus aureus. In some embodiments, the bacterial infection is caused by methicillin-resistant Staphylococcus aureus (MRSA).

[0554] In some embodiments, the bacterial infection is caused by gram-negative bacteria. The gram-negative bacteria include, but are not limited to, Escherichia species, Enterobacter species, Klebsiella species, Pseudomonas species, Acinetobacter species, Proteus species, Campylobacter species, Citrobacter species, Neisseria species, Salmonella species, Shigella species, Serratia species, Haemophilus species, Brucella species, Francisella species, Yersinia species, Burkholderia species, and Moraxella species.

[0555] In some embodiments, bacterial infection is caused by fermentation or non-fermentation Gram-negative bacteria.Examples of fermentation or non-fermentation Gram-negative bacteria include but are not limited to: Pseudomonas aeruginosa; Stenotrophomonas maltophilia; Burkholderia cepacia; Alcaligenes xylosoxidans; Enterobacteriaceae; Haemophilus; Franciscellaceae (e.g. Franciscellaceae); 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.

[0556] In some embodiments, the gram-negative bacterial infection is caused by the following infection: Escherichia species, Enterobacter species, Klebsiella species, Pseudomonas species, Acinetobacter species, Proteus species, Campylobacter species, Citrobacter species, Neisseria species, Salmonella species, Shigella species, Serratia species, Haemophilus species, Brucella species, Francisella species, Yersinia species, Burkholderia species or Moraxella species. In some embodiments, the gram-negative bacterial infection is caused by the infection of Enterobacteriaceae, Haemophilus, Freundii, or Neisseria species. In some embodiments, the bacterial infection is caused by Enterobacteriaceae. In some embodiments, the bacterial infection is caused by the Enterobacteriaceae that produces extended spectrum beta-lactamase (ESBL) or tolerates carbapenem (carbapenem).

[0557] In some embodiments, the gram-negative bacterial infection is an infection caused by: Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter kremerii, Proteus mirabilis, Bacillus anthracis, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia glanders, Burkholderia pseudomallei, or Yersinia pestis. In some embodiments, the gram-negative bacterial infection is an infection caused by: Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Burkholderia cepacia, or Alcaligenes xylosoxidans.

[0558] In some embodiments, the bacterial infection is caused by multidrug resistant (MDR) bacteria. In some embodiments, the bacterial infection is caused by multidrug resistant biological threat pathogens, including those defined by the A / B / C bioterrorism agent designation of the Centers for Disease Control and Prevention. In some such embodiments, the biological threat pathogen is a gram-negative bacterium.

[0559] In some embodiments, one or more compounds of Formula (I) or Formula (III) or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, or pharmaceutical compositions of the present disclosure can be used to treat diseases or conditions caused by bacterial infection disclosed herein, including but not limited to: skin infections; pneumonia; nosocomial pneumonia; sepsis; respiratory tract infections (e.g., lower or upper respiratory tract infections); lung infections in patients with cystic fibrosis; acute exacerbation of chronic bronchitis; community acquired pneumonia; Acquired pneumonia; nosocomial pneumonia (including ventilator-associated pneumonia (VAP)); hospital-acquired or community-acquired infections caused by gram-negative bacteria; upper airway disease; diffuse panbronchiolitis; tonsillitis; pharyngitis; acute sinusitis and otitis media, including mastoiditis; urinary tract and genital infections, such as complicated urinary tract infection, cystitis, resistant or recurrent cystitis, urethritis, pyelonephritis, acute pyelonephritis, endometritis, prostatitis, salpingitis, and epididymitis; eye infections, such as conjunctivitis, corneal ulcers, bloodstream infections, such as septicemia or bacteremia; skin and soft tissue infections, such as infected dermatitis, infected wounds, infected burns, phlegmon, folliculitis, and impetigo; bone and joint infections, such as osteomyelitis and septic arthritis; gastrointestinal infections, such as dysentery, enteritis, colitis, necrotizing enterocolitis, and anorectal infection;Intra-abdominal infections, including but not limited to complicated intra-abdominal infections, typhoid fever, infectious diarrhea, appendicitis with peritonitis, pelvic peritonitis and intra-abdominal abscesses; infections of the oral region, such as infections following dental surgery; other infections, such as melioidosis, endocarditis, sexually transmitted diseases caused by gram-negative or gram-positive bacteria, hepatic abscesses, cholecystitis, cholangitis, mastitis, and meningitis and nervous system infections.

[0560] In some embodiments, the methods disclosed herein treat an infection selected from the group consisting of complicated intra-abdominal infection, complicated urinary tract infection, drug-resistant or recurrent cystitis, acute pyelonephritis, and bacteremia.

[0561] In a further aspect, the present disclosure provides the use of one or more compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, for treating a bacterial infection in a subject in need thereof as described herein.

[0562] In a further aspect, the present disclosure provides the use of one or more compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, for treating a bacterial infection in a subject in need thereof.

[0563] In another aspect, the present disclosure provides one or more compounds of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer of any of the foregoing, for use as a medicament.

[0564] In a further aspect, the present disclosure provides the use of one or more compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, for the manufacture of a medicament for treating a bacterial infection in a subject in need thereof.

[0565] In a still further aspect, provided herein is the use of one or more pharmaceutical compositions of the present disclosure in a method of treating a bacterial infection in a subject in need thereof.

[0566] In a still further aspect, provided herein is the use of one or more pharmaceutical compositions of the present disclosure in the manufacture of a medicament for treating a bacterial infection in a subject in need thereof.

[0567] In any of the above methods and uses disclosed herein, the subject may be a mammal. In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human.

[0568] Dosage / Administration

[0569] One or more compounds of Formula (I) or Formula (III), or a pharmaceutically acceptable salt, solvate, stereoisomer or tautomer of any of them, or a composition of any of them, may be administered in a therapeutically effective amount, which will vary depending on various factors including: the activity of the specific compound employed; the metabolic stability and duration of action of the compound; the age, weight, general health, sex and diet of the subject; the mode and time of administration; the rate of excretion; drug combinations; the severity of the specific disease or condition; and the subject being treated.

[0570] Administration of one or more compounds of the present disclosure (e.g., compounds of Formula (I) or Formula (III)) or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers of any of them in pure form or in a suitable pharmaceutical composition can be carried out by any accepted mode of administration that exerts a similar effect. In some embodiments, the one or more compounds of the present disclosure administered are compounds of Formula (I) or Formula (III), or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers of any of the foregoing, administered in pure form or in a suitable pharmaceutical composition.

[0571] The pharmaceutical compositions can be used in the methods described herein and can be administered to a subject in need using techniques known to those of ordinary skill in the art, which techniques can be suitable as a therapy for a disease or condition affecting the subject. It will be understood by those of ordinary skill in the art that the dosage, duration of administration, and frequency of administration of the pharmaceutical compositions described herein to a subject in need depends on several factors, including, but not limited to: the subject's health status, the patient's specific disease or condition, the grade or level of the specific disease or condition, additional therapies that the subject is or has been using, etc.

[0572] The methods and compositions described herein can be used to administer to a subject in need thereof. Generally, administration of the compositions described herein can include routes of administration, non-limiting routes of administration include: oral, intravenous, intraarterial, subcutaneous, subdural, intramuscular, intracranial, intrasternal, intratumoral or intraperitoneal. In addition, the pharmaceutical composition can be administered to a subject by other routes of administration, for example, by inhalation, skin, intranasal or intrathecal administration.

[0573] Compositions of the present disclosure can be used in the first administration and in one or more additional administrations to objects in need thereof. The one or more additional administrations can be used to objects in need within a few minutes, hours, days, weeks or months after the first administration. Any of the additional administrations can be used to objects in need 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 be carried out more than once a day, more than once a week, or more than once a month. Administration can be weekly, every other week (every two weeks), every three weeks, every month or every two months.

[0574] One or more compounds of formula (I) or formula (III) or any of them, pharmaceutically acceptable salts, solvates, stereoisomers or tautomers, may also be administered simultaneously with, before, or after one or more other therapeutic agents. Such combination therapy may include administering a single pharmaceutical dosage formulation containing: one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, and one or more additional active agents, and administering one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, each active agent being in its own pharmaceutical dosage formulation. For example, one or more compounds of the present disclosure or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, and other active agents may be administered to a subject together in a single oral dosage composition (e.g., tablets or capsules), or each agent may be administered in a separate oral dosage formulation. When separate dosage forms are used, one or more compounds of the present disclosure, or pharmaceutically acceptable salts, solvates, stereoisomers or tautomers thereof, and one or more other active agents may be administered at substantially the same time (e.g., simultaneously), or at separately staggered times (e.g., sequentially); combination therapy is understood to include all of these regimens.

[0575] Reagent test kit

[0576] In some aspects, the present disclosure provides a kit comprising a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof, or a composition comprising the compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof, and instructions.

[0577] In some embodiments, the present disclosure provides a kit comprising a pharmaceutical composition as disclosed herein and instructions.

[0578] In some embodiments, the instructions include instructions for administering the disclosed compound, salt, or pharmaceutical composition to an object in need thereof. In some embodiments, the kit includes a compound, salt, or pharmaceutical composition disclosed herein, which is packaged in a low moisture vapor permeation container (transmission container) with a desiccant. Optionally, the label is on or associated with the container. For example, when letters, numbers, or other characters forming the label are attached, molded, or etched onto the container itself, the label may be located on the container; when it is present in a receptacle or carrier that also holds the container, the label may be associated with the container, such as as a package insert. In addition, the label can be used to indicate that the contents will be used for a specific therapeutic application. In some embodiments, the label includes instructions for use of the contents, such as in the methods described herein. In some embodiments, the compound, salt, or pharmaceutical composition disclosed herein is present in a package or container, which contains one or more unit dosage forms, and the unit dosage forms contain a compound, salt, or pharmaceutical composition disclosed herein. The package may include metal or plastic foil (such as a blister pack). The package or container may be accompanied by instructions for administration of the unit dosage form. In some embodiments, the package or container is accompanied by a notice in a form prescribed by a governmental agency that manages the manufacture, use or sale of drugs, which reflects the agency's approval of the drug form for human or veterinary administration. For example, such notices may be prescription drug labels approved by the U.S. Food and Drug Administration or approved product inserts. In some embodiments, a composition comprising a compound, salt, or pharmaceutical composition disclosed herein is prepared, placed in an appropriate container, and labeled to treat the indicated condition.

[0579] As used herein, a "kit" may include a container for containing at least one pharmaceutical composition or compound of the present disclosure, and may also include a divided container, such as a divided bottle or a divided foil packet. The container may be any conventional shape or form known in the art made of pharmaceutically acceptable materials, such as a paper or cardboard box, a glass or plastic bottle or jar, a resealable bag (e.g., for containing "refilled" tablets for placement in a different container), or a blister pack with individual doses to be pressed out of the package according to the treatment regimen. The container used may depend on the exact dosage form involved, for example, a conventional cardboard box is not usually used to hold a liquid suspension. It is feasible that more than one container can be used simultaneously in a single package to sell a single dosage form. For example, tablets can be contained in a bottle, and the bottle is contained in a box.

[0580] An example of such a kit is a so-called blister package. Blister packages are well known in the packaging industry and are widely used in the packaging of drug unit dosage forms (tablets, capsules, etc.). Blister packages are usually composed of a sheet of relatively hard material, which is covered with a foil of a transparent plastic material. During the packaging process, a recess is formed in the plastic foil. The recess has the size and shape of a single tablet or capsule to be packaged, or may have a size and shape to accommodate a plurality of tablets and / or capsules to be packaged. Subsequently, the tablet or capsule is placed in the recess accordingly, and the sheet of relatively hard material is sealed against the plastic foil on the face of the foil opposite to the groove formation direction. Therefore, the tablet or capsule is individually sealed or collectively sealed in the recess between the plastic foil and the sheet as required. In some embodiments, the strength of the sheet makes it possible to remove the tablet or capsule from the blister package by manually applying pressure to the recess, thereby forming an opening in the sheet at the recess. Subsequently, the tablet or capsule can be removed via the opening.

[0581] Another embodiment of the kit is a dispenser designed to dispense one daily dose at a time in the order of its intended use. In some embodiments, the dispenser is equipped with a memory-aid to further promote compliance with the regimen. An example of such a memory-aid is a mechanical counter that indicates the number of daily doses that have been dispensed. Another example of such a memory-aid is a battery-powered microchip memory coupled to a liquid crystal reader or an audible reminder signal, for example, to read out the date of administration of the last daily dose and / or to remind people when the next dose is to be taken.

[0582] In addition to one or more compounds or pharmaceutical compositions of the present disclosure, the kits of the present disclosure may also include 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 include a second antibacterial compound. The additional compound or pharmaceutical composition may be administered in the same dosage form as the one or more compounds or pharmaceutical compositions of the present disclosure, or in a different dosage form. Similarly, the additional compound or pharmaceutical composition may be administered simultaneously with the one or more compounds or pharmaceutical compositions of the present disclosure, or may be administered at different times.

[0583] In some embodiments, the kit of the present disclosure has a notice that is a notice required by a governmental regulatory agency that regulates the manufacture, use or sale of drugs or biological products, which reflects (a) approval for manufacture, use or sale for human administration, (b) instructions for use, or both.

[0584] Although described herein with reference to specific embodiments, it should be understood that these embodiments only illustrate the concept and application of the present disclosure. It is obvious to those skilled in the art that various modifications and changes can be made to the methods and devices of the present technology without departing from the spirit and scope of the present disclosure. Therefore, the present technology is intended to include modifications and changes within the scope of the attached claims and their equivalents. Therefore, the present disclosure is not limited except for the attached claims.

[0585] "One embodiment" or "an embodiment" mentioned in the specification means that the specific features, structures or characteristics described in the embodiment are included in at least one embodiment of the present disclosure. Therefore, the phrases "in one embodiment" or "in one embodiment" appearing in various places in this specification do not necessarily refer to the same embodiment. Moreover, the specific features, structures or characteristics can be combined in any suitable manner in one or more embodiments. Any ranges cited herein are inclusive.

[0586] Although preferred embodiments of the present invention are shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Those skilled in the art may make various changes, variations and substitutions without departing from the present invention. It should be understood that various alternative forms of the embodiments of the present invention described herein may be used to implement the present invention. The appended claims define the scope of the present invention, and methods and structures within the scope of these claims and their equivalents are encompassed by the present invention.

[0587] 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.

[0588] By reference to the following examples, aspects of the present technology can be more fully described. Before describing several exemplary embodiments of the present technology, it should be understood that the present technology is not limited to the details of the construction or process steps set forth in the following description. The present technology can have other embodiments and can be practiced or performed in various ways. The following examples are intended to illustrate certain aspects of the present technology and should not be construed as limitations of the present technology. It is understood that those skilled in the art may be able to manufacture these compounds by similar methods or by combining other methods known to those skilled in the art. It is also understood that those skilled in the art will be able to manufacture further compounds within the scope of the present disclosure in a similar manner as described below by using appropriate starting materials and modifying the synthetic route as needed. Typically, starting materials and reagents can be obtained from commercial suppliers, or synthesized according to sources known to those skilled in the art, or prepared as described herein.

[0589] The reagent / reactant names are given as given on commercial bottles or by IUPAC conventions ChemDraw 19.0 ( The compounds named salts (e.g., hydrochloride, acetate, sulfate) may contain more than one molar equivalent of acid.

[0590] Example

[0591] abbreviation

[0592] The following abbreviations are used herein.

[0593] Ac acetate or acetyl

[0594] Ac2O Acetic anhydride

[0595] AcOH Glacial acetic acid

[0596] AIBN Azobisisobutyronitrile

[0597] BF3·OEt2 Boron trifluoride etherate

[0598] Bn Benzyl

[0599] BnBr Benzyl bromide

[0600] Boc tert-Butyloxycarbonyl

[0601] BSA bovine serum albumin

[0602] Bu3SnH Tri-n-butyltin hydride

[0603] Cbz Carboxybenzyl

[0604] CCl3CN Trichloroacetonitrile

[0605] CsF Cesium fluoride

[0606] d Double

[0607] DAST Diaminosulfur trifluoride

[0608] DCM Dichloromethane

[0609] dd double doublet

[0610] DIAD Diisopropyl azodicarboxylate

[0611] DMAP N,N-Dimethylaminopyridine

[0612] DMEM Dulbecco's Modified Ehrlich Medium

[0613] DMF N,N-Dimethylformamide

[0614] DMSO dimethyl sulfoxide

[0615] DMTMM chloride 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride

[0616] DPPA Diphenylphosphoryl azide

[0617] equiv equivalent

[0618] EtOAc Ethyl acetate

[0619] Et2O Ether

[0620] EtOH

[0621] g

[0622] h hour

[0623] HPLC High Pressure Liquid Chromatography

[0624] LAH Lithium Aluminum Hydride

[0625] LC / MS Liquid Chromatography / Mass Spectrometry

[0626] LiOH Lithium Hydroxide

[0627] m multiplet

[0628] M Moore

[0629] MeCN Acetonitrile

[0630] MeOH Methanol

[0631] min

[0632] mmol millimole

[0633] MPLC Medium Pressure Liquid Chromatography

[0634] MS

[0635] n-normal (unbranched)

[0636] N Positive / Normal

[0637] NaH Sodium hydride

[0638] NaN3 Sodium Azide

[0639] NaOMe Sodium Methanol

[0640] NBS N-Bromosuccinimide

[0641] NH4OAc Ammonium acetate

[0642] NMR Nuclear Magnetic Resonance

[0643] Pd(OH)2 Palladium Hydroxide

[0644] pH-log[H + ]

[0645] PhI(OAc)2 Phenyliodonium acetate

[0646] Piv Pivaloyl (trimethylacetyl)

[0647] PMB p-Methoxybenzyl

[0648] PMB-Cl p-Methoxybenzyl chloride

[0649] ppm parts per million

[0650] psi pounds per square inch

[0651] PTSA p-Toluenesulfonic acid

[0652] RP-HPLC Reversed Phase High Pressure Liquid Chromatography

[0653] R t Retention time

[0654] rt room temperature

[0655] s single peak

[0656] tTriple Peak

[0657] t-uncle

[0658] t-BuOH tert-Butanol

[0659] TBAI Tetra-n-butylammonium iodide

[0660] TBSCl tert-Butyldimethylsilyl chloride

[0661] TEA triethylamine

[0662] TfN3 trifluoromethanesulfonyl azide

[0663] TFA Trifluoroacetic acid

[0664] THF Tetrahydrofuran

[0665] TMS Trimethylsilyl

[0666] TMSOTf trimethylsilyl trifluoromethanesulfonate

[0667] TolSH p-Toluenethiophenol

[0668] TsCl p-Toluenesulfonyl chloride

[0669] USP United States Pharmacopeia

[0670] wt% weight percentage / weight%

[0671] Analytical methods

[0672] The analytical data are included in the following procedures. Unless otherwise stated, all 1 H NMR data were all collected on a Bruker Ultra Shield 500 MHz / 54 mm instrument (BZH 43 / 500 / 70B, D221 / 54-3209) and chemical shifts are expressed in parts per million (ppm).

[0673] Example 1. Synthesis of (benzyl(((2S,5R)-5-azido-6-hydroxytetrahydro-2H-pyran-2-yl)methyl)(benzyl 1-(4-(4-(4-yl)carbamate) (ring A1)

[0674]

[0675] Step A. (S)-N-((E)-((S)-3,4-dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinyl Amide

[0676]

[0677] (S)-(+)-2-methyl-2-propanesulfinamide (50g, 412mmol) and CuSO4 (131.7g, 825mmol) are mixed in DCM (600.0mL). During 20 minutes, a solution of 2-formyl-3,4-dihydro-2H-pyran (48.6g, 433mmol, Synthonix company) in DCM (100.0mL) is added. The mixture is stirred at room temperature for 18h, then filtered through diatomaceous earth, and rinsed with DCM. The filtrate is evaporated under reduced pressure. Use 0-20% Et2O in hexane solution as eluent, by MPLC on silica gel (5x330g, dry load) upper purified material, obtain title compound (23.74g, 27%, the diastereomer of the second elution). 1 H NMR (400MHz, CDCl3) δ8.08 (d, J = 3.0 Hz, 1H), 6.47 (dt, J = 6.3, 1.6 Hz, 1H), 4.81-4.72 (m, 2H), 2.19-1.94 (m, 4H), 1.24 (s, 9H).

[0678] Step B. (R)-N-(((S)-3,4-dihydro-2H-pyran-2-yl)methyl)-2-methylpropane-2-sulfenamide

[0679]

[0680] To a solution of (NE)-N-[[(2S)-3,4-dihydro-2H-pyran-2-yl]methylene]-2-methyl-propane-2-sulfinamide (572 mg, 2.66 mmol) in ethanol (10.0 mL) at 0 ° C. Sodium borohydride (101 mg, 2.66 mmol) was added. The ice bath was removed and the reaction was kept stirring for another 30 minutes. The reaction was cooled to 0 ° C. and saturated NH4Cl (20.0 mL) was added (note: gas is produced). EtOH was evaporated under reduced pressure, and the residue was extracted with EtOAc (30.0 mL). The layers were separated, and the organic phase was washed with water (10.0 mL) and brine (10.0 mL) in sequence, the organic phase was dried (Na2SO4), filtered and concentrated under reduced pressure to give the title compound as a solid (547 mg, 95%). LCMS (m / z): ES + [M+H] + :218.20; (A05) retention time = 2.04m. This material was used in the subsequent step without further purification.

[0681] Step C. (R)-N-Benzyl-N-[[(2S)-3,4-dihydro-2H-pyran-2-yl]methyl]-2-methyl-propane-2-yl Sulfenamide

[0682]

[0683] To a mixture of (R)-N-[[(2S)-3,4-dihydro-2H-pyran-2-yl]methyl]-2-methyl-propane-2-sulfinamide (547 mg, 2.52 mmol) and BnBr (448 μL, 3.78 mmol) in DMF (1.5 mL) at 0 ° C. Sodium hydride (60%, 116 mg, 2.89 mmol) was added. The mixture was stirred at room temperature for 1 hour, then cooled to 0 ° C., followed by the addition of water (10 mL). The aqueous phase was extracted with EtOAc (25.0 mL), and the organic phase was washed successively with water (10.0 mL) and brine (5.0 mL), and the organic phase was dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (24 g column) using EtOAc and hexane (0-40%) to give the title compound (768 mg, 99%) as an oil. LCMS m / z: ES + [M+H] + :308.18; (A05) retention time = 2.57m.

[0684] Step D. (R)-N-Benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-2-methyl 2-Propane-2-sulfenamide

[0685]

[0686] To a suspension of (R)-N-benzyl-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (320 mg, 1.04 mmol) and NaHCO (262 mg, 3.12 mmol) in 1:1 H2O / ACN (8.0 mL) at ambient temperature was added iodine (272 mg, 1.07 mmol). After 30 minutes, MeCN was evaporated and the residue was partitioned between EtOAc (20.0 mL) and 1:1 saturated NaHCO3 / saturated Na2S2O3 (10.0 mL). The organic layer was washed with water (5.0 mL) and brine (5.0 mL), dried (Na2SO4), filtered and concentrated under reduced pressure to give the title compound as a foam (4 diastereomers, 450 mg, 96%). LCMS m / z:ES + [M+H] + :452.06; (A05) retention time = 2.30-2.50m. This material was used in the subsequent step without further purification.

[0687] Step E. (R)-N-[[(2S)-5-Azido-6-oxo-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl- Propane-2-sulfenamide

[0688]

[0689] To a solution of benzyl (R)-N-benzyl-N-[[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]methyl]-2-methyl-propane-2-sulfenamide (450 mg, 997 μmol) in dry DCM (10.0 mL) at ambient temperature under N2 was added molecular sieves ( 500mg) and pyridinium dichromate (750mg, 1.99mmol). After 24h, the solution was concentrated and the crude product was distributed between EtOAc (30.0mL) and water (15.0mL). The organic layer was washed with water (2x10.0mL), brine (10.0mL), dried (Na2SO4) and concentrated. The crude product was dissolved in dry DMF (1.5mL) under N2, and NaN3 (130mg, 1.99mmol) was added. After 1h, the solution was distributed between EtOAc (30.0mL) and water (15.0mL). The organic layer was washed with water (2x10.0mL) and brine (10.0mL), the organic layer was dried (Na2SO4) and concentrated to give the title compound (235mg, 64%) as an oil. LCMS m / z:ES + [M+H] + :365.17; (A05) retention time = 2.37m. This material was used in the subsequent step without further purification.

[0690] Step F. (R)-N-[[(2S,5R)-5-Azido-6-hydroxy-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl 2-Propane-2-sulfenamide

[0691]

[0692] To a solution of (R)-N-[[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl-propane-2-sulfinamide (235 mg, 645 μmol) in dry DCM (8.0 mL) at -78 ° C under N2 was added a solution of DIBAL-H in toluene (1M, 1.03 mL, 1.03 mmol) dropwise. After 100 minutes, saturated potassium sodium tartrate (15.0 mL) was slowly added to the solution, followed by water (15.0 mL). The mixture was warmed to room temperature and stirred for 16 hours. The mixture was extracted with DCM (10.0+2x5.0 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (12 g column) using EtOAc and hexanes (10%-50%) to give the title compound (diastereomers, 195 mg, 74%) as an oil. LCMS m / z: ES + [M+H] + :367.19; (A05) retention time = 2.28-2.40min.

[0693] Step G. Benzyl N-[[(2S,5R)-5-azido-6-hydroxytetrahydropyran-2-yl]methyl]-N-benzyl-amino Formate (Ring A1)

[0694]

[0695] To a solution of (R)-N-[[(2S)-5-azido-6-hydroxy-tetrahydropyran-2-yl]methyl]-N-benzyl-2-methyl-propane-2-sulfinamide (195 mg, 532 μmol) in dioxane (2.7 mL) was added aqueous HCl (1.0 M, 2.66 mL, 2.66 mmol) dropwise with vigorous stirring. After 1 hour, solid NaCO (451 mg, 4.26 mmol) was added. After another 5 minutes, CbzCl (106 μL, 745 mmol) was added dropwise. After another 1 hour, the dioxane was evaporated and the residue was partitioned between EtOAc (20.0 mL) and H2O (20.0 mL). The organic phase was washed with brine (10.0 mL), dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude product was purified by C18 reverse phase chromatography (80 g column) using ACN and 0.1% aqueous formic acid (40-70%) to give the title product (mixture of 4 diastereomers) as an oil (167 mg, 79%). UPLC m / z: ES+ [M-OH] + :379.21; (CSH 5 to 100% ACN / ammonium formate pH 4) retention times = 2.12, 2.15, 2.20, and 2.22 m. A portion (40 mg) of the material was purified by preparative HPLC (CSH C18 ACN / ammonium formate 50-70%) to provide the α-azide as a film (20 mg, 50% recovery). UPLC m / z: (CSH 5 to 100% ACN / ammonium formate pH 4) retention times = 2.15 and 2.22 min.

[0696] Example 2. Synthesis of ((1S)-1-((2S,5R)-5-azido-6-hydroxytetrahydro-2H-pyran-2-yl)ethyl) Benzyl (Benzyl)carbamate (Ring A2)

[0697]

[0698] Step A. ((S)-N-((E)-((S)-3,4-dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-yl Sulfonamide

[0699]

[0700] (S)-(+)-2-methyl-2-propanesulfinamide (50g, 412mmol) and CuSO4 (131.7g, 825mmol) are mixed in DCM (600.0mL). During 20 minutes, a solution (100.0mL) of 2-formyl-3,4-dihydro-2H-pyran (48.6g, 433mmol, Synthonix company) in DCM is added. The mixture is stirred at room temperature for 18h, then filtered through diatomaceous earth, and rinsed with DCM. The filtrate is evaporated under reduced pressure. Using 0-20% Et2O in hexane solution as eluent, material is purified by MPLC on silica gel (5x330g, dry load) to obtain the title compound (23.74g, 27%, the diastereomer of the second elution). 1 H NMR (400MHz, CDCl3) δ8.08 (d, J = 3.0 Hz, 1H), 6.47 (dt, J = 6.3, 1.6 Hz, 1H), 4.81-4.72 (m, 2H), 2.19-1.94 (m, 4H), 1.24 (s, 9H).

[0701] Step B. (S)-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinyl Amide

[0702]

[0703] (S)-N-((E)-((S)-3,4-dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinamide (7.30 g, 33.9 mmol) was dissolved in dry DCM (100 mL), and the solution was cooled to -78 ° C. MeMgBr (3.0 M in Et2O, 22.6 mL, 67.8 mmol) was added dropwise to the mixture, and stirring was continued at low temperature for 2 hours, after which the mixture was allowed to warm to room temperature. Stirring was continued at room temperature for 1 hour. Concentrated NH4Cl aqueous solution (50 mL) was added, and the mixture was extracted with DCM (3x50 mL). The combined organic layers were dried (MgSO4) and concentrated under vacuum to give a crude oil containing a mixture of diastereoisomers. The material was purified using silica gel chromatography (120 g column) with hexane and ethyl acetate (0-100%) to give the title compound (4.40 g, 56%) as an oil. LCMS m / z:ES + [M+H] + :232.16.

[0704] Step C. (S)-N-Benzyl-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane Alkane-2-sulfenamide

[0705]

[0706] To a solution of (S)-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide (4.40 g, 19.0 mmol) and benzyl bromide (3.39 mL, 28.5 mmol) in DMF (100 mL) was added NaH (60% dispersion, 913 mg, 22.8 mmol) at 0 ° C. The mixture was stirred at this temperature for 2 hours, followed by the addition of brine (100 mL) at 0 ° C. The aqueous layer was extracted with Et2O (3x50.0 mL). The combined organic layers were dried (MgSO4) and concentrated under vacuum to give a residue that was purified using silica gel chromatography (120 g column) using hexane and ethyl acetate (0-100%). The pure fractions were concentrated under vacuum to give the title compound (3.50 g, 57%) as an oil. LCMS m / z:ES + 322.14.

[0707] 1H NMR(500MHz,DMSO)δ7.45(d,J=7.1Hz,2H),7.35-7.29(m,2H),7.23(t,J=7.3Hz,1H ),6.38(d,J=6.1Hz,1H),4.65(ddd,J=6.2,4.3,1.4Hz,1H),4.35(d,J=16.5Hz,1H) ,3.91(t,J=11.4Hz,1H),3.80(td,J=9.2,2.3Hz,1H),3.22-3.11(m,1H),1.97-1.9 2(m,1H),1.89-1.78(m,2H),1.53-1.42(m,1H),1.19(t,J=3.4Hz,3H),1.10(s,8H).

[0708] Step D. (S)-N-Benzyl-N-((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)- 2-Methylpropane-2-sulfenamide

[0709]

[0710] (S)-N-Benzyl-N-((S)-1-((S)-3,4-dihydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide (4.40 g, 13.7 mmol) and NaHCO (3.45 g, 41.1 mmol) were dissolved in H2O (75.0 mL) and ACN (75.0 mL). I2 (3.82 g, 15.1 mmol) was added portionwise at 0 °C for 10 min. The cooling bath was removed and stirring was continued at room temperature for 90 min. The reaction mixture was quenched with a saturated aqueous solution of Na2S2O3 (30.0 mL) and extracted with EtOAc (3x20.0 mL). The combined organic layers were dried (MgSO4) and concentrated under vacuum to give the title mixture as a solid (4.70 g, 74%). LCMS m / z: ES+[M+H]+: 466.04.

[0711] Step E. Benzyl((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)carbamate ester

[0712]

[0713] (S)-N-Benzyl-N-((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)-2-methylpropane-2-sulfinamide (4.70 g, 10.1 mmol) was dissolved in 1,4-dioxane (150 mL) and 1N HCl (25.2 mL, 25.2 mmol) aqueous solution was added. Stirring was continued at room temperature for 20 minutes, after which solid NaCO (8.56 g, 80.8 mmol) was added to the mixture. Stirring was continued for 20 minutes, after which CbzCl (1.72 mL, 12.1 mmol) was added dropwise. Stirring was continued for 2 hours. Water (200 mL) was added, and the mixture was extracted with EtOAc (3x50 mL). The combined organic layers were dried (MgSO4) and concentrated in vacuo to give a residue which was purified by silica gel chromatography (80 g column) with hexanes and ethyl acetate (0-100%). Pure fractions were concentrated in vacuo to give the title compound (3.70 g, 74%) as an oil. LCMS m / z: ES+[M+H]+: 518.04.

[0714] Step F. ((1S)-1-((2S)-5-azido-6-oxotetrahydro-2H-pyran-2-yl)ethyl)(benzyl)amino Benzyl formate

[0715]

[0716] Benzyl ((1S)-1-((2S)-6-hydroxy-5-iodotetrahydro-2H-pyran-2-yl)ethyl)carbamate (3.70 g, 7.47 mmol) was dissolved in DCM (350 mL). Suspend the mixture Molecular sieves (1.00 g) were added, and PDC (8.43 g, 22.4 mmol) was added. Stirring was continued at room temperature for 24 hours, then filtered through a pad of celite using ethyl acetate and concentrated under vacuum to give crude benzyl ((1S)-1-((2S)-5-iodo-6-oxotetrahydro-2H-pyran-2-yl)ethyl)carbamic acid benzyl ester as an oil. The crude product was dissolved in DMF (50.0 mL). Subsequently, NaN3 (971 mg, 14.9 mmol) was added as a solid, and stirring was continued at room temperature for 2 hours. The solvent was removed under vacuum, and the residue was dissolved in EtOAc (100 mL) and washed with water (3x25 mL). The combined organic layers were dried (MgSO4) and concentrated under vacuum to give an oil, which was purified by silica gel chromatography (80 g column) using ethyl acetate and hexanes (0-100%). The pure fractions were combined and concentrated in vacuo to give the title compound as an oil (1.40 g, 46%). LCMS m / z: ES + [M+Na] + :431.13; calculated:431.18.

[0717] Step G. ((1S)-1-((2S,5R)-5-azido-6-hydroxytetrahydro-2H-pyran-2-yl)ethyl)(benzyl)amine Benzyl carboxylate (ring A2)

[0718]

[0719] Benzyl ((1S)-1-((2S)-5-azido-6-oxotetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (1.40 g, 3.43 mmol) was dissolved in dry DCM (75.0 mL) and the mixture was cooled to -78 °C. A solution of DIBAL-H (1.0 M in toluene, 6.86 mL, 6.86 mmol) was added to the mixture and stirring was continued at low temperatures for 1 hour. EtOH (2.00 mL) was added dropwise to the cold reaction mixture and then poured into a saturated aqueous solution of Rochelle salt (200 mL). The mixture was stirred vigorously for 1 hour and then extracted with EtOAc (3x50.0 mL). The combined organic layers were dried (MgSO4) and concentrated under vacuum to give a residue which was purified using silica gel chromatography (80 g column) using hexane and ethyl acetate (0-70%). Pure fractions were collected and concentrated in vacuo to give the title compound as a mixture of diastereomers (800 mg, 57%). LCMS m / z: ES + [M+Na] + :433.16; Calculated:433.19.

[0720] Example 3. Synthesis of N-[(1R)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N- Benzyl-carbamic acid benzyl ester (ring A3)

[0721]

[0722] Step A. (R)-N-((E)-((S)-3,4-dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinyl Amide

[0723]

[0724] (R)-(+)-2-methyl-2-propanesulfinamide (81.1g, 669mmol) and CuSO4(213.5g, 1.34mol) are mixed in DCM (700mL).During 20 minutes, a solution (100mL) of 2-formyl-3,4-dihydro-2H-pyrans (75g, 669mmol) in DCM is added.The mixture is stirred at room temperature for 72h, then filtered through diatomite, and rinsed with DCM.The filtrate is evaporated under reduced pressure.Use 0-10% Et2O hexane solution as eluent, by MPLC on silica gel (5x330g, dry load) upper purified material, obtain the title compound (28g, 19%) in solid. 1H NMR (400MHz, CDCl3) δ8.07 (d, J = 3.1 Hz, 1H), 6.40 (d, J = 5.8 Hz, 1H), 4.76-4.68 (m, 1H), 4.68-4.63 (m, 1H), 2.13-1.83 (m, 4H), 1.18 (s, 9H).

[0725] Step B N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinyl Amide

[0726]

[0727] To a solution of (R)-N-((E)-((S)-3,4-dihydro-2H-pyran-2-yl)methylene)-2-methylpropane-2-sulfinamide (7.10 g, 33.0 mmol) in dry DCM (110 mL) was added MeMgBr (3.0 M in EtO, 22.0 mL, 66.0 mmol) at -40 °C under N2. After 2 hours, the reaction was allowed to warm to room temperature within 1 h. After another 14 h, saturated NH4Cl (160 mL) was added dropwise (caution: gas was produced). The two phases were separated and the aqueous phase was extracted with DCM (3x20.0 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (220 g column) using EtOAc and hexanes (20-45%) to give the title compound (6.00 g, 79%) as a solid. 1 H NMR (500MHz, CDCl3) δ6.37 (d, J=6.1Hz, 1H), 4.75-4.61 (m, 1H), 3.93 (ddd, J=1 1.2,3.3,2.0Hz,1H),3.69(d,J=6.1Hz,1H),3.56(pd,J=6.7,3.5Hz,1H),2.16- 2.06(m,1H),1.99(dtt,J=17.1,5.6,1.6Hz,1H),1.81(ddd,J=13.3,6.6,1.8Hz ,1H),1.66(dtd,J=13.3,11.4,5.8Hz,1H),1.22(s,9H),1.18(d,J=6.7Hz,3H).

[0728] Step C. (R)-N-Benzyl-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl- Propane-2-sulfenamide

[0729]

[0730] At 0 ° C, to a mixture of (R)-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl] ethyl]-2-methyl-propane-2-sulfinamide (1.03 g, 4.44 mmol) and BnBr (0.79 mL, 6.67 mmol) in DMF (60 mL) was added NaH (60%, 187 mg, 4.89 mmol). The mixture was stirred at room temperature for 1 h, and then brine (250 mL) was added at 0 ° C. The aqueous phase layer was extracted with Et2O (3x80 mL). The combined organic solution was dried over MgSO4 and concentrated under reduced pressure. Using hexane to 60% EtOAc in hexane, the material was purified by MPLC on silica gel (40 g, dry load) to obtain the title compound (1.2 g, 84%) as an oil. 1 H NMR (400MHz, cdcl3) δ7.37(d,J=7.5Hz,2H),7.32(t,J=7.4Hz,2H),7.26(d,J=3.8Hz,1H),6.32(d,J=6.2Hz,1H),4.64(s,1H),4.39(d,J=15.3H z,1H),4.14(d,J=15.2Hz,1H),3.66-3.57(m,1H),3.32-3.20(m,1H),1. 98-1.80(m,3H),1.54-1.43(m,1H),1.37(d,J=6.8Hz,3H),1.19(s,9H).

[0731] Step D. (R)-N-Benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-2-methyl 2-Propane-2-sulfenamide

[0732]

[0733] To a suspension of (R)-N-benzyl-N-[(1R)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (1.2 g, 3.73 mmol) and NaHCO (941 mg, 11.2 mmol) in ACN (50 mL) and H O (50 mL) was added I (947 mg, 3.73 mmol) in portions at 0 ° C. The mixture was stirred at room temperature for 1 hour, followed by the addition of a saturated aqueous solution of Na S O (10 mL). The aqueous layer was extracted with EtOAc (3 x 150 mL). The combined organic layers were washed with brine, dried over MgSO, filtered and concentrated under reduced pressure to give the title compound (1.6 g, 92%) as a solid. M+H + :466.1.

[0734] Step E-N-Benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]carbamic acid Benzyl ester

[0735]

[0736] To (R)-N-benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (1.2 g, 2.58 mmol) in dioxane (40 mL) was added 1N HCl (6.45 mL, 6.45 mmol). The mixture was stirred at room temperature for 20 minutes, followed by the addition of Na2CO3 (2.19 g, 20.6 mmol). After 20 minutes, CbzCl (0.51 mL, 3.61 mmol) was added dropwise. The mixture was stirred at room temperature for 2 hours. Water (200 mL) was added. The separated aqueous phase was extracted with EtOAc (3x150 mL). The combined organic layers were washed with brine, dried over MgSO4, and concentrated under reduced pressure. The material was purified by MPLC on silica gel (80 g, dry loaded) using hexanes to 80% EtOAc to give the title compound as a solid (1.1 g, 86%). M+H + :496.0.

[0737] Step F N-[(1R)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-benzyl-amino Benzyl formate

[0738]

[0739] To a solution of N-benzyl-N-[(1R)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]carbamic acid benzyl ester (1.10 g, 2.22 mmol) in DCM (100 mL) at 0°C was added Dess-Martin Periodinane (1.88 g, 4.44 mmol). The mixture was stirred at room temperature for 5 hours. Water (100 mL) was added, followed by a saturated aqueous solution of Na2S2O3. The separated aqueous phase was extracted with DCM (2x50 mL). The combined organic layers were washed with a saturated aqueous solution of NaHCO3 (2x100 mL), brine (100 mL), dried over MgSO4, and concentrated under reduced pressure. The residue was then taken up in anhydrous DMF (75 mL), and NaN3 (217 mg, 3.33 mmol) was added. The mixture was stirred at room temperature for 15 minutes, followed by the addition of brine (300 mL). The aqueous layer was extracted with Et2O (3x100 mL). The combined organic layers were dried over MgSO4 and concentrated under reduced pressure. The material was purified by MPLC on silica gel (40 g, dry loaded) using hexanes to EtOAc to give the title compound (800 mg, 88%) as an oil. M+H + :409.3.

[0740] Step GN-[(1R)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl- Benzyl carbamate (ring A3)

[0741]

[0742] To a solution of N-[(1R)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (800 mg, 1.96 mmol) in DCM (60 mL) was added a solution of DIBAL-H (1M in toluene, 11.8 mL, 11.8 mmol) dropwise at -78 ° C. After 1 h at -78 ° C, EtOH (0.5 mL) was added dropwise. The mixture was poured into a saturated aqueous solution of Rochelle salt (300 mL). The mixture was vigorously stirred for 1 hour. The separated aqueous phase was extracted with DCM (2x75 mL). The combined organic layers were washed with brine, dried over MgSO4, and concentrated under reduced pressure. The residue was purified by MPLC on silica gel (40 g, dry load) using hexane to 60% EtOAc in hexane to give the title compound (363 mg, 41%) as an oil. M+H + :411.2.

[0743] Example 4. Synthesis of (1R,2R,3S,4R,5S,6S)-3,5-bis(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexane Alkane-1,2,4-triyl triacetate (ring B1)

[0744]

[0745] Step A: Synthesis of (1S,2R,3S,4S,5R,6S)-3,5-diamino-6-(((2R,3R,4R,5S)-3-(((2R,3S,4S,5R,6S)-4,5-dihydroxy-6-(hydroxymethyl)-3-(methylamino)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-4-(hydroxymethyl)-5-methyltetrahydrofuran-2-yl)oxy)cyclohexane-1,2,4-triol

[0746]

[0747] Dihydrostreptomycin sulfate (50g), Ba (OH) -8H o (4 equivalents) and water (500mL) are merged and heated 2 days under reflux.After being cooled to room temperature, by CO blowing through reaction, until this reaction becomes slightly acidic (pH 6-7).The gained mixture is filtered through diatomite, washed with water twice (75mL).Under vacuum, the aqueous layer is concentrated, and in the situation of not further purification, the aqueous solution will be directly used.

[0748] 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

[0749]

[0750] During ~1 hour, at room temperature, Cbz-OSu (57.7 g, 3.4 eq.) in THF (110 mL) was added to the crude compound from step A. The reaction was stirred at room temperature for 2 days. A saturated aqueous solution of sodium bicarbonate (50 mL) was added, and the product was extracted into EtOAc (2x75 mL) and washed with brine (50 mL). The combined organics were dried over sodium sulfate and the solvent was removed under vacuum. The crude product was dissolved in 2-butanone (80 ml) and slowly dripped into MTBE (750 mL). The precipitated solid was collected by filtration and dried in a vacuum oven at 50 ° C to give the title product (52.1 g) as an off-white solid (purity 72% determined by absorption at 210 nm).

[0751] 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

[0752]

[0753] The crude compound (52.1 g) from step B was dissolved in EtOAc (210 mL) and DMAP (721 mg, 0.1 equivalent) was added at room temperature. At room temperature, DIPEA (103 mL) and Ac2O (51 mL) in EtOAc (80 mL) were added dropwise to the reaction mixture during 1 h. The reaction was stirred overnight at room temperature. The reaction was then transferred to a separatory funnel with EtOAc (50 mL, 0.9 V) and washed with water (50 mL), sodium bicarbonate solution (2x50 mL), and brine (50 mL). The organic matter was dried over sodium sulfate and filtered to give the title product (70 g; 63% AP at 210 nm) as a creamy foam.

[0754] Step D: Synthesis (1R,2R,3S,4R,5S,6S)-3,5-Bis(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexane Alkane-1,2,4-triyl triacetate (ring B1)

[0755]

[0756] The crude product from step C is added to DCM (420mL), dodecanethiol (29.4mL, 2.1 equivalents) and BF3.Et2O (16.3mL, 2.2 equivalents). The reaction is heated for 2 hours under reflux. The reaction is washed with sodium bicarbonate aqueous solution, water and brine (each 20mL). The organic portion is concentrated under vacuum, and the resulting solid is vigorously stirred with MeCN / hexane (1:2, 210mL) to obtain the title product. The crude product is ground with heptane (300mL) to obtain the title product (23.9g, 71% yield).

[0757] Example 5. Synthesis of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butyric acid (F-HABA)

[0758]

[0759] Step A: Synthesis of (R)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyacetic acid methyl ester

[0760]

[0761] To a solution of D-isoascorbic acid (100 g, 0.568 mol, 1.0 eq.) and 2,2-dimethoxypropane (76.8 mL, 0.624 mol, 1.1 eq.) in dry acetone (500 mL) was slowly added concentrated H2SO4 (0.6 mL, 11.2 mmol) and the mixture was stirred at 0°C for 5 h. To the reaction mixture was slowly added NaHCO3 (142.8 g, 1.7 mol, 3.0 eq.) and H2O (400 mL). Acetone was removed by rotary evaporator under reduced pressure. To the residue was slowly added 30% H2O2 (128.4 mL, 1.132 mol, 2.0 eq.) using a dropping funnel at 0°C. After the mixture was stirred for 3 h, Na2SO3 (14.28 g, 113.2 mmol, 0.2 eq.) was added and the mixture was stirred for 30 min. Sodium bicarbonate (192.8 g, 2.28 mol, 4.0 equivalents) and dimethyl sulfate (225.6 mL, 2.36 mol, 4.0 equivalents) were added and stirred at 50 ° C for 5 h. The reaction mixture was filtered and the aqueous phase layer (800 mL × 2) was extracted with CH2Cl2. The organic layers were combined and dried over anhydrous MgSO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography, eluted with petroleum ether / ethyl acetate (2: 1, v / v) to obtain (R)-2-((R)-2,2-dimethyl-1,3-dioxolane-4-yl)-2-hydroxyacetic acid methyl ester (70 g, 64.8%) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ4.35-4.14(m,2H),4.01-3.90(m,2H),3.75(s,3H),3.03(d,J=6.4Hz,1H),1.36(s,3H),1.29(s,3H).

[0762] Step B: Synthesis of (S)-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethane-1,2-diol

[0763]

[0764] To a suspension of LAH (16.59 g, 0.44 mol, 1.2 eq.) in dry THF (600 mL) was added dropwise a solution of (R)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyacetic acid methyl ester (70 g, 0.36 mol, 1.0 eq.) in THF (200 mL) under N. The resulting mixture was stirred at room temperature for 3 hours, then H2O (20 mL) was dripped dropwise, followed by addition of 15% NaOH aqueous solution (20 mL) and H2O (60 mL). The reaction mixture was filtered and concentrated, and the residue was purified by flash column chromatography, eluting with petroleum ether / ethyl acetate (1:1) to give (S)-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethane-1,2-diol (45 g, 75.4% yield). 1 H NMR (400MHz, DMSO-d6) δ4.21(br,s,2H),3.79-3.95(m,3H),3.29-3.50(m,3H),1.30(s,3H),1.25(s,3H).

[0765] Step C: Synthesis of (S)-2-((R)-2,2-dimethyl-1,3-dioxolane-4-yl)-2-hydroxyethyl 4-methylbenzenesulfonate

[0766]

[0767] To a solution of (S)-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethane-1,2-diol (45 g, 277.8 mmol, 1.0 equiv) in dry pyridine (500 mL) was added TsCl (52.96 g, 277.8 mmol, 1.0 equiv) portionwise at 0°C over 1 h. The resulting mixture was stirred at room temperature overnight, and then the solvent was removed. The residue was dissolved in EA (1000 mL), washed with saturated aqueous NH4Cl (500 mL×2), dried over MgSO4, filtered, and concentrated. The residue was purified by flash column chromatography using petroleum ether / ethyl acetate (4:1) as eluent to give (S)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyethyl 4-methylbenzenesulfonate (59 g, 75.9% yield). 1H NMR (400MHz, DMSO) δ7.79(d,J=8.2Hz,2H),7.49(d,J=8.1Hz,2H),5.53(d,J=5.9Hz,1H),4.11-4.03(m,1H), 3.98-3.85(m,2H),3.80(td,J=10.0,4.9Hz,2H),3.56-3.44(m,1H),2.42(s,3H),1.21(s,3H),1.19(s,3H).

[0768] Step D: Synthesis of (S)-2-azido-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethan-1-ol

[0769]

[0770] To a solution of (S)-2-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2-hydroxyethyl 4-methylbenzenesulfonate (59 g, 186.6 mmol, 1.0 equiv) in dry DMF (500 mL) was added NaN3 (36.38 g, 559.7 mmol, 3.0 equiv) at room temperature. The mixture was warmed to 80°C and stirred overnight. The resulting mixture was poured into a saturated aqueous solution of Na2CO3 (2000 mL) and extracted with ethyl acetate (1000 mL×2). The combined organic phases were washed with a saturated aqueous solution of NH4Cl (1000 mL×2), dried over MgSO4, filtered and concentrated. The residue was purified by flash column chromatography using petroleum ether / ethyl acetate (8:1) to give (S)-2-azido-1-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)ethan-1-ol (34 g, 97.4% yield). 1 H NMR (400MHz, DMSO) δ5.54(d,J=5.8Hz,1H),3.99(dd,J=7.7,5.9Hz,1H),3.92-3.79(m,2H),3.53( ddd,J=8.3,6.5,2.8Hz,1H),3.32(m,1H),3.22(dd,J=12.8,6.5Hz,1H),1.30(s,3H),1.25(s,3H).

[0771] Step E: Synthesis of (R)-4-((R)-2-azido-1-fluoroethyl)-2,2-dimethyl-1,3-dioxolane

[0772]

[0773] To a solution of (S)-2-azido-1-((R)-2,2-dimethyl-1,3-dioxolane-4-yl)ethane-1-ol (34 g, 0.18 mol, 1.0 equiv) in dry DCM (170 mL) was added a solution of DAST (73 mL, 0.54 mol, 3.0 equiv) dropwise at -20 ° C, and the reaction mixture was then allowed to warm to room temperature and stirred overnight. The resulting mixture was slowly poured into a saturated aqueous solution of NaHCO3 (1500 mL) and extracted with DCM (500 mL×2), and the combined organic phases were dried over MgSO4, filtered and concentrated. The residue was purified by flash column chromatography, eluting with petroleum ether / ethyl acetate (30:1) to give (R)-4-((R)-2-azido-1-fluoroethyl)-2,2-dimethyl-1,3-dioxolane (3.4 g, 9.9% yield). 1 H NMR (400MHz, CDCl3) δ4.57(ddt,J=48.0,7.4,3.6Hz,1H),4.24(dtd,J=21.4,6.7,3.6Hz,1H),4.14-4.05(m,1H),3.94(dd ,J=8.4,6.5Hz,1H),3.63(ddd,J=17.0,13.5,7.5Hz,1H),3.44(ddd,J=27.4,13.5,3.6Hz,1H),1.44(s,3H),1.38(s,3H). 19 F NMR (400MHz, CDCl3) δ198.91 (s, 1F).

[0774] Step F: Synthesis of (2R,3R)-4-azido-3-fluorobutane-1,2-diol.

[0775]

[0776] To a solution of (R)-4-((R)-2-azido-1-fluoroethyl)-2,2-dimethyl-1,3-dioxolane (3.4 g, 18.0 mmol, 1.0 equiv) in THF (30 mL) was added 10% HCl (4 mL) dropwise at 0°C. The mixture was allowed to warm to room temperature and stirred overnight. The organic solvent was removed, and the residue was basified to pH = 9 with a saturated aqueous solution of Na2CO3, extracted with EA (40 mL×4), dried over MgSO4, filtered, and concentrated. The residue was purified by flash column chromatography using petroleum ether / ethyl acetate (1:1) as eluent to give (2R,3R)-4-azido-3-fluorobutane-1,2-diol (2.5 g, yield: 93.3%). 1H NMR (400MHz, DMSO) δ5.17 (d, J = 5.9 Hz, 1H), 4.86 (t, J = 5.5 Hz, 1H), 4.74 (ddt, J = 48.8, 8.4, 2.9 Hz, 1H), 3.79-3.44 (m, 5H). 19 F NMR (400MHz, DMSO-d6) δ202.16 (s, 1F).

[0777] Step G: Synthesis of (2R,3R)-4-azido-1-((tert-butyldimethylsilyl)oxy)-3-fluorobutan-2-ol.

[0778]

[0779] To a solution of (2R,3R)-4-azido-3-fluorobutane-1,2-diol (2.5 g, 16.7 mmol, 1.0 eq) in DCM (50 mL) was added DMAP (4.51 g, 36.9 mmol, 2.2 eq) at 0°C, followed by portionwise addition of TBS-Cl (2.77 g, 18.5 mmol, 1.1 eq). The reaction mixture was allowed to warm to room temperature and stirred for 3 days. The organic solvent was evaporated and the residue was purified by flash chromatography eluting with petroleum ether / ethyl acetate (10:1) to give (2R,3R)-4-azido-1-((tert-butyldimethylsilyl)oxy)-3-fluorobutan-2-ol (2.6 g, yield: 59.1%). 1 H NMR (400MHz, DMSO) δ5.17 (d, J = 5.7Hz, 1H), 4.71-4.50 (m, 1H), 3.73-3.37 (m, 5H), 0.82 (s, 9H), 0.00 (s, 6H). 19 F NMR (400MHz, DMSO-d6) δ203.48 (s, 1F).

[0780] Step H: Synthesis of ((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butoxy)(tert-butyl)dimethylsilane

[0781]

[0782] To a solution of (2R,3R)-4-azido-1-((tert-butyldimethylsilyl)oxy)-3-fluorobutan-2-ol (2.6 g, 9.88 mmol, 1.0 eq) in THF (250 mL) was added NaH (474 ​​mg, 11.86 mmol, 1.2 eq, 60%) portionwise at 0°C. After stirring at 0°C for 30 minutes, TBAI (3.64 g, 9.88 mmol, 1.0 eq) was added, followed by PMB-Cl (2.0 mL, 14.73 mmol, 1.5 eq) dropwise at 0°C. The reaction was allowed to warm to room temperature and stirred overnight. The mixture was poured into a saturated aqueous solution of NH4Cl and extracted with EA (50 mL×3). The extract was dried over MgSO4, filtered and concentrated. The residue was purified by flash column chromatography using petroleum ether / ethyl acetate (40:1) to give ((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butoxy)(tert-butyl)dimethylsilane (2.5 g, yield: 66.1%). 1 H NMR (400 MHz, dioxane) δ 8.31 (d, J = 8.5 Hz, 2H), 7.95 (d, J = 8.6 Hz, 2H), 5.90-5.69 (m, 1H), 5.58 (dd, J = 40.1, 11.2 Hz, 2H), 4.79 (s, 3H), 4.78-4.48 (m, 5H), 1.91 (s, 9H), 1.09 (s, 6H). 19 FNMR(400MHz,DMSO-d6)δ199.15(s,1F).

[0783] Step I: Synthesis of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butan-1-ol

[0784]

[0785] To a solution of ((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butoxy)(tert-butyl)dimethylsilane (2.5 g, 6.48 mmol, 1.0 equiv) in DMSO (15 mL) was added CsF (2.98 g, 19.44 mmol, 3.0 equiv) followed by 5 drops of MeOH. The mixture was stirred at room temperature overnight and then poured into saturated NaCl (40 mL), extracted with EA (40 mL×3), and the extract was dried over MgSO4, filtered and concentrated. The residue was purified by flash column chromatography using petroleum ether / ethyl acetate (4:1) as eluent to give (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butane-1-ol (1 g, yield: 56.8%). 1H NMR (400MHz, DMSO) δ7.35-7.20(m,2H),7.01-6.83(m,2H),5.76(s,3H),4.93(t,J=5.3Hz,1H),4.74(ddd,J=25.8,8 .0,5.6Hz,1H),4.52(dd,J=49.2,11.3Hz,2H),3.74(s,3H),3.69(ddd,J=18.8,13.9,8.1Hz,1H),3.58-3.42(m,4H). 19 FNMR(400MHz,DMSO-d6)δ199.45(s,1F).

[0786] Step J: Synthesis of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid (F-HABA)

[0787]

[0788] To a solution of (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butan-1-ol (1 g, 3.76 mmol, 1.0 eq) in ACN / H2O (10 mL / 10 mL) was added PhI(OAc)2 (2.99 g, 9.41 mmol, 2.5 eq) followed by TEMPO (292 mg, 1.88 mmol, 0.5 eq). The mixture was stirred at room temperature overnight. Sodium hydroxide (2N, 10 mL) was added and the mixture was extracted with PE (20 mL×4) and the extract was discarded. The aqueous phase was acidified to pH 3 with 2N HCl and extracted with EtOAc (30 mL x 4). The extract was dried over Na2SO4 and concentrated. The residue was purified by flash column chromatography using DCM / MeOH (20:1, v / v) to give (2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butanoic acid (500 mg, yield: 47.5%). 1 H NMR(400MHz,DMSO)δ7.30(d,J=8.7Hz,2H),6.93(d,J=8.7Hz,2H),5.11-4.86(m,1H),4.52 (dd,J=119.7,11.0Hz,2H),4.19(dd,J=31.5,2.6Hz,1H),3.76(s,3H),3.75-3.65(m,2H). 19 F NMR (400MHz, DMSO-d6) δ196.81 (s, 1F).

[0789] Example 6. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R, 4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl) oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl 2-Hydroxy-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutyramide (Compound 1)

[0790]

[0791] Step A: Synthesis of (E)-2,2,2-trifluoro-N-phenylacetimidate (3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl ester

[0792]

[0793] A mixture of N-[[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]methyl]-N-benzyl-carbamic acid benzyl ester (Example 1; 1.1 g, 2.50 mmol), Cs2CO3 (2.44 g, 7.49 mmol), and (1E)-2,2,2-trifluoro-N-phenyl-acetimidoyl chloride (1.2 mL, 7.49 mmol) in DCM (20 mL) was stirred at room temperature for 18 h. The mixture was filtered, rinsed with DCM, and concentrated under reduced pressure. The material was purified by silica gel chromatography (80 g column) using a gradient of 0 to 30% EtOAc in hexanes to give the title product as a syrup (1.2 g, 85%). m / z (ESI) [M+Na] + 590.3, HPLC (A05) tR = 3.02 min.

[0794] Step B: Synthesis of (1S,2S,3R,4S,5S,6R)-6-(((3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-3,5-bis(((benzyloxy)carbonyl)amino)cyclohexane-1,2,4-triyl triacetate

[0795]

[0796] [(3R,6S)-3-azido-6-[[benzyl(benzyloxycarbonyl)amino]methyl]tetrahydropyran-2-yl](1E)-2,2,2-trifluoro-N-phenyl-acetimidate (1.2 g, 2.12 mmol) and [(1S,2R,3S,4S,5R,6R)-3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (Ring B1; Example 4; 1.816 g, 3.17 mmol) were co-evaporated with anhydrous toluene (2x5 mL). The mixture was dried under reduced pressure for 30 minutes. The residue was taken up in anhydrous DCM (100 mL) and then transferred via cannula to a column filled with pre-activated and sieved RBF. The mixture was stirred at room temperature for 20 minutes and then cooled to -25 ° C. Bi (OTf) 3 (1.526g, 2.32mmol) was added once, and the mixture was stirred at -25 ° C for 20 minutes and stirred at room temperature for 2h. The mixture was diluted with saturated NH4Cl solution (50mL) and DCM (100mL), then filtered (diatomaceous earth). The separated aqueous layer was extracted with DCM (3x50mL). The combined organic layer was washed with brine (20mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse phase chromatography using a gradient of 5 to 100% ammonium formate (10mM, pH 4) in MeCN, and the material was freeze-dried to obtain the title product (1.22g, 59%) as a mixture of anomers (α / β=~2.7:1). m / z (ESI) [M+H] + :951.5, HPLC tR=2.82 min.

[0797] Step C: Synthesis of dibenzyl ((1R,2R,3S,4R,5R,6S)-4-(((3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2,5,6-trihydroxycyclohexane-1,3-diyl)dicarbamate

[0798]

[0799] To a suspension of [(1S,2R,3S,4S,5R,6S)-3,4-diacetoxy-5-[(3R,6S)-3-azido-6-[[benzyl(benzyloxycarbonyl)amino]methyl]tetrahydropyran-2-yl]oxy-2,6-bis(benzyloxycarbonyl-amino)cyclohexyl]acetate (1.22 g, 1.28 mmol) in MeOH (30 mL) was added K2CO3 (887 mg, 6.41 mmol). The suspension was stirred at room temperature for 80 minutes and then filtered. The filtrate was concentrated under reduced pressure (bath temperature <30°C) to give a residue. Water (15 mL) was added to form a precipitate, which was sonicated, filtered and washed with water (50 mL). The solid was collected, dissolved in DCM / MeOH (50 mL, 10:1), dried (MgSO4), filtered, and concentrated under reduced pressure to give the title product as a solid (1.06 g, crude, quantitative). m / z (ESI) [M+H] + ; 825.4, HPLC A(05)tR=2.63 min.

[0800] Step D: Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarbamate

[0801]

[0802] A mixture of N-[[(2S,5R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy-cyclohexyloxy]tetrahydropyran-2-yl]methyl]-N-benzyl-carbamic acid benzyl ester (1 g, 1.21 mmol), 1,1-dimethoxycyclohexane (1.11 mL, 7.27 mmol) and PPTS (122 mg, 0.485 mmol, dried on V10 via high boiling method before use) in dry DME (30 mL) was stirred at 90° C. for 2 h. The mixture was cooled and then concentrated under reduced pressure. The material was purified by reverse phase column on ACCQ using 65-87% ammonium formate (pH 4) in MeCN to give the title compound as a solid (633 mg, 58%, α anomer) β anomer (250 mg, 23%). m / z: ES+[M+H] + 905.1; LCMS (A05); tR = 2.99 min (α anomer).

[0803] Step E: Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarbamate

[0804]

[0805] To a mixture of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarbamate (630 mg, 0.644 mmol) and BnBr (0.94 mL, 7.85 mmol) in THF (20 mL) at 0°C under nitrogen was added NaH (60% oil dispersion, 64.5 mg, 1.62 mmol). The mixture was stirred at 0°C for 3 h, then AcOH (0.1 mL, 1.29 mmol) and EtOAc (30 mL) were added. The mixture was concentrated under reduced pressure. The material was purified by silica gel chromatography (40 g column) using a gradient of 0 to 50% EtOAc in hexanes to give the title compound as a solid (630 mg, 91%). m / z: ES+[M+H] + 995.5; LCMS (A05); tR = 3.13 min.

[0806] Step F: Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate

[0807]

[0808] A solution of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarbamate (630 mg, 0.633 mmol) in THF (12 mL), water (3.5 mL) and AcOH (8 mL) was heated at 80 °C for 9 h and then cooled to room temperature. The mixture was diluted by slowly adding saturated aqueous NaHCO3 (~50 mL) at 0 °C and stirred for 20 minutes. The separated aqueous layers were extracted with EtOAc (3x50 mL) each. The combined organic layers were washed with saturated aqueous NaHCO3 (3x10 mL), water (3x10 mL), brine (2x10 mL), dried (MgSO4), filtered and concentrated under reduced pressure to give the title product as an off-white solid (570 mg, 98%). The material was used directly without further purification. m / z (ESI) [M+H] + :915.4, HPLC (A05) tR=2.85 min.

[0809] Step G: Synthesis of benzyl (((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)methyl)(benzyl)carbamate (ring AB1)

[0810]

[0811] To a solution of dibenzyl ((lS,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate (570 mg, 0.623 mmol) in dry DMF (9 mL) at 0°C was added sodium hydride (60% oil dispersion, 125 mg, 3.11 mmol). The mixture was stirred at 0°C for 3 h then diluted with AcOH (0.28 mL, 4.83 mmol) and concentrated under reduced pressure. The material was purified by silica gel chromatography (40 g column) using a gradient of 0 to 100% EtOAc in hexanes to give the title compound (440 mg, 88% yield) as a solid. m / z(ESI)[M+H] +:807.4, HPLC (A05) tR=2.77 min.

[0812] Step H: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((3aR,4S,5S,6R,7S,7aS)-6-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-oxooctahydroxybenzo[d]oxazol-7-yl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0813]

[0814] To a mixture of [(2R, 3R, 4R)-4-acetoxy-3-[(2R, 3R, 4R, 5R, 6S)-4, 5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-5-hydroxy-tetrahydrofuran-2-yl]methyl acetate (ring CD1, International Patent Application Publication No. WO2019194858, incorporated by reference; 1157 mg, 2.18 mmol) and K2CO3 (452.4 mg, 3.272 mmol) in dry DCM (15 mL) was added CCl3CN (0.328 mL, 3.27 mmol). The mixture was stirred at room temperature for 18 h, then filtered on celite, rinsed with CH2Cl2, and concentrated under reduced pressure. To the previous crude material was added N-[[(2S,5R,6R)-6-[[(3aR,4S,5S,6R,7S,7aS)-4-benzyloxy-5-(benzyloxycarbonylamino)-7-hydroxy-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-6-yl]oxy]-5-azido-tetrahydropyran-2-yl]methyl]-N-benzyl-carbamic acid benzyl ester (Cycle AB1; 440 mg, 0.545 mmol) in dry DCM (30 mL) and then transferred to a column filled with pre-activated and The mixture was stirred at room temperature for 1 h and then diluted with a saturated aqueous solution of NaHCO (15 mL). The mixture was filtered through diatomaceous earth and the solid was rinsed with DCM (200 mL). The separated aqueous phase was extracted with DCM (3x20 mL). The combined organic layers were washed with brine (15 mL), dried (MgSO ), filtered, and concentrated under reduced pressure. The material was purified by a reverse phase column (ACCQ) using a gradient of 64%-85% MeCN (10 mM, pH 4) in ammonium acetate to obtain the title product (455 mg, 63%). m / z (ESI) [M+H] + :1319.5, HPLC (A05) tR=2.95 min.

[0815] Step I: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-amino-6-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0816]

[0817] To the preparation of [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6[[benzyl(benzyloxycarbonyl)amino]methyl]-tetrahydropyran-2-yl]oxy-4-benzyloxy-5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-7-yl]acetate To a solution of 1,4-dioxane (12 mL) and water (6 mL) was added LiOH HO (434 mg, 10.35 mmol). The mixture was stirred at 40 ° C for 18 h, then cooled to room temperature and diluted with EtOAc (60 mL). The separated aqueous layer was extracted with EtOAc (3x20 mL), respectively. The combined organic layers were washed with water (3x5 mL), brine (10 mL), dried (MgSO4), filtered and concentrated under reduced pressure to give the title compound (382 mg, crude, 98%) as an off-white solid. The material was used directly without further purification. m / z(ESI)[M+H] + :1125.5, HPLC (A05) tR=2.43 minutes.

[0818] Step J: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butyramido)-6-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0819]

[0820] To N-[[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy- [cyclohexyloxy]-5-azido-tetrahydropyran-2-yl]methyl]-N-benzyl-carbamic acid benzyl ester (180mg, 0.160mmol) and (2R, 3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butyric acid (F-HABA, Example 5; 77.7mg, 0.274mmol) in anhydrous THF (12mL) were added DMTMM chloride (77.7mg, 0.281mmol). The mixture was stirred at room temperature for 18h, then diluted with NH4Cl saturated aqueous solution (5mL) and DCM (30mL). The separated aqueous layer was extracted with DCM (3x10mL) respectively. The combined organic layer was washed with brine (5mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse phase column using a 10 to 100% MeCN gradient in 10 mM ammonium formate (pH 3.8) and lyophilized to give the title compound as a white solid (187 mg, 84%). m / z (ESI) [M+H] + :1390.6, HPLC A(05)tR=2.85min.

[0821] Step K: Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (Compound 1)

[0822]

[0823] To a suspension of Pd(OH)2 (20% wet, 652 mg, 0.93 mmol) in a mixture of AcOH (4.8 mL) and water (2 mL) was added N-[[(2S,5R,6R)-5-azido-6-[(1R,2R,3S,4S,5R,6S)-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydro- A solution of 4-[[(2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butyryl]amino]-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy-cyclohexyloxy]tetrahydropyran-2-yl]methyl]-N-benzyl-carbamic acid benzyl ester (187 mg, 134 μmol) in MeOH (6 mL). The mixture was degassed by 3x vacuum-H2 cycles and then bubbling with hydrogen for 10 minutes. The mixture was stirred under a hydrogen atmosphere (1 atm) for 3 h and then degassed with N2 for 5 min. The suspension was filtered on a 0.40 μM PVDF filter, then washed with MeOH (5x8 mL) and water (5x10 mL), and the volatiles were evaporated under reduced pressure (rotary evaporation bath temperature <35 ° C). 10mM NH is used by preparative HPLC4HCO3 in 5 to 15% MeCN (pH 10) is purified by material, and lyophilized to obtain solid (100mg). At 0°C, H2SO4 (0.05M in H2O) is added dropwise to the solution of the above solid in H2O (3mL), until pH 4, then lyophilized. The solid is dissolved in water (2mL), and MeOH (10mL) is added to obtain precipitation. The precipitation is then filtered through a PVDF syringe filter, and washed with MeOH (2x5mL). Mother liquor is discarded, and the solid is dissolved (5mL) with water, and the solution is lyophilized to obtain the title product compound 1 (67.2mg, 49%) as an off-white solid. 1 H NMR(400MHz,D2O)δ5.77(d,J=3.5Hz,1H),5.24(d,J=1.9Hz,1H),5.22-5.02( m,2H),4.47-4.35(m,1H),4.33-4.24(m,2H),4.21-4.15(m,1H),4.14-3.94(m ,4H),3.89-3.74(m,3H),3.72-3.55(m,3H),3.55-3.47(m,1H),3.47-3.04(m, 8H), 2.91 (dd, J=13.4, 8.2Hz, 1H), 2.01-1.69 (m, 3H), 1.44 (t, J=12.6Hz, 1H). Note: 26 exchangeable protons not observed.

[0824] Example 7. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R,4S, 5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy (((2R,3R,6S)-3-amino-6-(aminomethyl)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl) Tetrahydro-2H-pyran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutyramide (Compound 2)

[0825] (Compound 2).

[0826] Step A: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0827]

[0828] (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-amino-6-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy ... To a solution of (2S)-2-benzyloxy-4-(benzyloxycarbonylamino)butanoic acid (HABA; 101 mg, 0.293 mmol), and DIPEA (0.10 mL, 0.587 mmol) in anhydrous DMF (4 mL) was added PyBOP (122 mg, 0.235 mmol). The mixture was stirred at room temperature for 1.5 h and poured into water (5 mL) and EtOAc (5 mL). The separated aqueous layers were extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse phase column (ACCQ) using ACN and ammonium formate (10 mM, pH 3.8) and lyophilized to give the title compound (142 mg, 50%) as a white solid.

[0829] M+H+:1451.7

[0830] Step B: Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-4-(((2R,3R,6S)-3-amino-6-(aminomethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 2)

[0831]

[0832] To (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino To a solution of (474 ​​mg, 0.67 mmol, 20% wet) in a mixture of AcOH (5.0 mL) and water (2.0 mL) was added Pd(OH)2 (474 ​​mg, 0.67 mmol, 20% wet) in AcOH (5.0 mL) and water (2.0 mL). The mixture was bubbled with hydrogen for 10 minutes. The mixture was stirred under a hydrogen atmosphere (1 atm) for 3 h and then degassed with N2 for 5 min. The suspension was filtered on a 0.22 μM PPVDF filter syringe and washed with MeOH (3 x 3 mL) and water (2 x 2 mL). The volatiles were evaporated under reduced pressure (temperature < 35 °C). Use 10mM NH4HCO35% MeCN (pH 10) by filtration to purify the material, and freeze-dry to obtain a solid. At 0 ° C, to the above solid in H2O (3mL) solution dropwise add H2SO4 (0.05M in H2O), until pH 4-5, then freeze-dried. The solid is dissolved in water (2mL), then MeOH (10mL) is added to obtain a precipitation. The solid is filtered through a 0.22uM PVDF syringe filter, rinsed with MeOH (2x5mL), then washed with MeOH (5mL). The solid is dissolved in water (10mL), then freeze-dried to obtain the title compound 2 (58mg, 60%) as a white solid. 1H NMR(400MHz,D2O)δ5.96(d,J=3.6Hz,1H),5.44(s,1H),5.33(s,1H),4.62-4.54(m,1H),4.50-4.45 (m,1H),4.45-4.35(m,2H),4.30-4.12(m,4H),4.03-3.90(m,3H),3.88-3.76(m,3H),3.69(t,J=9. 8Hz,1H),3.65-3.55(m,2H),3.51-3.40(m,2H),3.39-3.25(m,2H),3.25-3.16(m,2H),3.11(dd,J= 13.5,8.2Hz,1H),2.30-2.17(m,1H),2.14-2.02(m,3H),1.97(d,J=14.7Hz,1H),1.68-1.58(m,1H).

[0833] Example 8. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R, 6S)-3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4- (((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutyramide (compound 3)

[0834]

[0835] Step A: Synthesis of (E)-2,2,2-trifluoro-N-phenylacetimidate (3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl ester

[0836]

[0837] To a solution of 2,2,2-trifluoro-N-phenyl-acetimidyl chloride (10.4 mL, 64.9 mmol) and N-[(1S)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (Ring A2; Example 2, Step G; 14.8 g, 36.1 mmol) in DCM (336 mL) at 23° C. under nitrogen was added cesium carbonate (23.5 g, 72.2 mmol). The mixture was stirred at 23° C. for 45 minutes, filtered through a pad of celite, rinsed with DCM (200 mL), and concentrated. The residue was purified by silica gel chromatography (330 g high capacity column) using EtOAc and hexanes (0-20%) to give the title compound (21 g, quantitative) as a solid. m / z: ES+ no ionization; LCMS (A05) tR = 3.14 min.

[0838] Step B: Synthesis of (1S,2S,3R,4S,5S,6R)-6-(((3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-3,5-bis(((benzyloxy)carbonyl)amino)cyclohexane-1,2,4-triyl triacetate

[0839]

[0840] [(3R, 6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl](1E)-2,2,2-trifluoro-N-phenyl-acetimidate 2 (21.5 g, 37 mmol) and [(1S, 2R, 3S, 4S, 5R, 6R)-3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (ring B1; 31.7 g, 55.5 mmol) were co-evaporated with anhydrous toluene (2x45 mL). The mixture was dried under reduced pressure for 30 minutes. The residue was taken up in anhydrous DCM (550 mL) and EtO (250 mL) and then transferred to a RBF filled with pre-activated 4A sieves via a syringe. The mixture was stirred at room temperature for 30 minutes, then Bi(OTf)3 (26.7 g, 40.7 mmol) was added at once at 0°C. The mixture was stirred at room temperature for 3h, then diluted with saturated aqueous NH4Cl solution (500 mL) and DCM (400 mL). The suspension was filtered through a celite pad, and the layers were partitioned. The separated aqueous layer was extracted with DCM (3x300 mL). The combined organic layers were washed with brine (500 mL), dried (MgSO4), filtered and concentrated under reduced pressure to give a crude product 3 (56 g crude product) as a mixture of β / α=1:9 anomers. The material was used in the next step without further purification. m / z:ES+[M+H] + 965.3; LCMS (A05); tR = 3.00 min (β anomer), 3.06 min (α anomer).

[0841] Step C: Synthesis of N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy-cyclohexyloxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester

[0842]

[0843] To a solution of [(1S,2S,3R,4S,5S,6R)-2-acetoxy-3-[(3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy-4,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (56 g, 58 mmol, crude) in MeOH (375 mL) was added sodium methoxide (4.60 M in MeOH, 10 mL, 46.4 mmol) at room temperature. The mixture was stirred at room temperature for 1 h, diluted with AcOH (10 mL) until pH ~ 5, and then concentrated under reduced pressure. The residue was purified by preparative HPLC (X-bridge C18, 50x250 mm, 10 μM) using 58-72% ACN (10 mM, pH 3.8) of ammonium formate. The fractions were pooled and concentrated under reduced pressure to remove the ACN. The aqueous residue was extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (2 x 100 mL), dried (MgSO4), filtered and concentrated under reduced pressure to give the title compound as a solid (17.5 g, 59% after 3 steps). m / z (ESI) [M+H] + ; 839.3, HPLC A(05)tR=2.80 min.

[0844] Step D: Synthesis of N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-5,7-bis(benzyloxycarbonylamino)-6-hydroxy-spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxol-2,1'-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester

[0845]

[0846] To a solution of N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy-cyclohexyloxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (17.5 g, 20.9 mmol) and 1,1-dimethoxycyclohexane (26.9 mL, 177 mmol) in DME (337 mL) was added dried PPTS (2.17 g, 8.62 mmol). The mixture was stirred at 90 °C under nitrogen for 3 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified on silica gel (350 g, HC 25 uM) using EtOAc and hexane (0-100%) to give the title compound (17.5 g, 91%) as a white solid. m / z:ES+,[M+H] + 919.4; LCMS (A05); tR = 3.12 min.

[0847] Step E: Synthesis of N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-6-benzyloxy-5,7-bis(benzyloxycarbonylamino)spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxol-2,1'-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester

[0848]

[0849] To a mixture of benzyl N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-5,7-bis(benzyloxycarbonylamino)-6-hydroxy-spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxol-2,1'-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamate (18.5 g, 20.1 mmol) and benzyl bromide (26.8 mL, 226 mmol) in THF (500 mL) was added sodium hydride (60% dispersion in mineral oil, 1.85 g, 46.3 mmol) at 0 °C under nitrogen. The mixture was stirred at room temperature for 6 h (less than 50% conversion) and transferred to a 30 ° C oil bath, stirred for 5 h (complete conversion), followed by the addition of AcOH (2.27 mL, 39.7 mmol), followed by silica gel and EtOAc (150 mL). The mixture was concentrated and purified by silica gel chromatography (350 g column, 25 uM) using a gradient of 0-100% EtOAc in hexane. The mixed fractions were pooled, concentrated, and repurified by silica gel chromatography (350 g column) using a gradient of 0-50% EtOAc in hexane to give the title compound (16.8 g, 83%) as a solid. m / z: ES+[M+H] + 1009.4; LCMS (A05); tR = 3.23 min.

[0850] Step F: Synthesis of dibenzyl ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarbamate

[0851]

[0852] To a solution of N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4R,5S,6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy-cyclohexyloxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (17.5 g, 20.9 mmol) and 1,1-dimethoxycyclohexane (26.9 mL, 177 mmol) in DME (337 mL) was added dried PPTS (2.17 g, 8.62 mmol). The mixture was stirred at 90 °C under nitrogen for 3 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel (350 g, HC 25 uM) using EtOAc and hexane (0-100%) to give the title compound (17.5 g, 91%) as a white solid. m / z:ES+,[M+H] + 919.4; LCMS (A05); tR = 3.12 min.

[0853] Step G: Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate

[0854]

[0855] A solution of N-[(1S)-1-[(2S,5R,6R)-6-[(3aS,4R,5S,6S,7R,7aS)-6-benzyloxy-5,7-bis(benzyloxycarbonylamino)spiro[3a,4,5,6,7,7a-hexahydro-1,3-benzodioxole-2,1'-cyclohexane]-4-yl]oxy-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (16.8 g, 16.6 mmol) in THF (180 mL), water (60 mL) and AcOH (150 mL) was heated at 80 ° C for 12 h and then cooled to room temperature. The mixture was slowly added to water (2 L). The resulting precipitate was filtered, and the solid was washed with water (3x100 mL), collected and dissolved in EtOAc (500 mL). The solution was washed with a saturated aqueous solution of NaHCO3 (100 mL) and water (3x100 mL). The aqueous solution was extracted with EtOAc (3x50 mL). The combined organic phases were washed with brine (2x50 mL), dried (MgSO4), filtered and concentrated under reduced pressure to give the title product as a solid (14.8 g, 96%). The solid was used directly without further purification. m / z (ESI) [2M+NH4]+ :1875.3, HPLC (A05) tR=2.99 min.

[0856] Step H: Synthesis of benzyl ((S)-1-((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydroxybenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (ring AB2)

[0857]

[0858] To a solution of N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2S,3R,4S,5S,6R)-3-benzyloxy-2,4-bis(benzyloxycarbonylamino)-5,6-dihydroxy-cyclohexyloxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (14.8 g, 15.9 mmol) in dry DMF (150 mL) at 0°C was added sodium hydride (60% oil dispersion, 3.19 g, 79.7 mmol). The mixture was stirred at 0°C for 1 h30, diluted with AcOH (7.8 mL, 135 mmol) and then slowly poured into stirring water (1 L). The precipitate was filtered and the solid was washed with water (3 x 200 mL) and lyophilized to give the title product as a solid (13.8 g, >99%). The solid was further purified by silica gel column using a gradient of 0-100% EtOAc in hexanes to give the title product as a solid (11 g, 84%). m / z (ESI) [M+H] + :821.3, HPLC (A05) tR=2.90 min.

[0859] Step I: [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy-4-benzyloxy-5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-7-yl]oxy]-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-tetrahydrofuran-2-yl]methyl acetate

[0860]

[0861] activation

[0862] 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 CD-1 (6.9 g, 13 mmol) and CClCN (2 mL, 19.7 mmol) in dry DCM (300 mL) was added KCO (2.73 g, 19.7 mmol). The mixture was stirred at room temperature for 18 h, then filtered on Celite, rinsed with DCM, and concentrated under reduced pressure to give a slurry.

[0863] Glycosylation

[0864] A mixture of the slurry and N-[(1S)-1-[(2S,5R,6R)-6-[[(3aR,4S,5S,6R,7S,7aS)-4-benzyloxy-5-(benzyloxycarbonylamino)-7-hydroxy-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-6-yl]oxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (2.7 g, 3.29 mmol) was co-evaporated with dry toluene (12 mL) under reduced pressure, dissolved in dry DCM (300 mL) and then transferred to a RBF filled with 4A sieves. The mixture was cooled to -78°C and then BF3.OEt2 (6.12 mL, 49.6 mmol) was slowly added at -78°C over 5 minutes. Once the addition is complete, the dry ice bath is removed immediately. The mixture is stirred at room temperature for 2h, and by NaHCO saturated aqueous solution (150mL) and DCM (300mL) dilution. The mixture is filtered through Celite, and the filter cake is washed with DCM (3x150mL). The mixture is distributed, and the aqueous solution is extracted with DCM (3x50mL). The combined organic layer is washed with saline (2x100mL), dried (MgSO ), filtered, and concentrated under reduced pressure. The material is purified by preparation HPLC (X-bridge C18, 50x250mm, 10 μM) using 72-86% ACN (10mM, pH 3.8) of ammonium formate. The product fractions are collected, and ACN is removed under reduced pressure. The aqueous layer is extracted with EtOAc (3x150mL). The combined organic layers were washed with brine (2 x 50 mL), dried (MgSO4), filtered, and concentrated under reduced pressure to afford the title compound [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxyacetic acid 4-benzyloxy-5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzoxazol-7-yl]oxy]-4-acetoxy-3-[(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-tetrahydrofuran-2-yl]methyl ester (4 g, 91%). m / z(ESI)[M+H] + :1334.2, HPLC (A05) tR=3.04 min.

[0865] Step J: Synthesis of N-[(1S)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy-cyclohexyloxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester

[0866]

[0867] [(2R,3R,4R,5S)-5-[[(3aR,4S,5S,6R,7S,7aS)-6-[(2R,3R,6S)-3-azido-6-[(1S)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy-4-benzyloxy-5-(benzyloxycarbonylamino)-2-oxo-3a,4,5,6,7,7a-hexahydro-3H-1,3-benzo[1,2-dihydro-2-yl]-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl]oxy]- To a solution of [(2R,3R,4R,5R,6S)-4,5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-tetrahydrofuran-2-yl]methyl ester (4.0 g, 3.00 mmol) in dioxane (100 mL) and water (100 mL) was added LiOH HO (3.78 g, 90.0 mmol). The mixture was stirred at 40 ° C for 18 h, cooled to room temperature, and diluted with EtOAc (300 mL) and HO (50 mL), followed by partitioning. The separated aqueous layer was extracted with EtOAc (3x100 mL), respectively. The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure to give the title compound (3.40 g, 99%) as a white solid. m / z (ESI) [M+H] + :1139.4, HPLC A(05)tR=2.54min.

[0868] Step K: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butyramide)-6-(((2R,3R ,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0869]

[0870] To N-[(1S)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxy)- To the 4-nitro-2-nitro-1-yl)-4-nitro-2-yl-2-[(4-methoxyphenyl) methoxy]-4-nitro-2-yl-4-nitro-2-yl-3-fluoro-2-[(4-methoxyphenyl) methoxy]-N-benzyl-carbamic acid benzyl ester (3.2g, 2.81mmol) and (2R, 3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl) methoxy] butyric acid (1.37g, 4.82mmol) in anhydrous THF (150mL) solution add DMTMM chloride (1.37g, 4.93mmol). The mixture is stirred at room temperature for 18h, then by NH4Cl saturated aqueous solution (100mL) and EtOAc (300mL) dilution. The isolated aqueous layer is extracted with EtOAc (50mL x3) respectively. The combined organic layer is washed with brine (2x50mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by preparative HPLC (X-bridge C18, 50x250mm, 10μM) using 70-85% ACN (10mM, pH 3.8) of ammonium formate. The product fractions were pooled and concentrated under reduced pressure to remove the ACN. The aqueous solution was extracted in EtOAc (3x150mL). The combined organic phases were washed with brine (2x50mL), dried (MgSO4), filtered and concentrated under reduced pressure to give the title compound (2.7g, 68%) as a solid. m / z (ESI) [M+H] +:1404.4, HPLC A(05)tR=2.96min.

[0871] Step L: Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (Compound 3)

[0872]

[0873] A mixture of MeOH (152 mL), AcOH (127 mL), and water (51 mL) was transferred via syringe under N2 to a round-bottom flask charged with N-[(1S)-1-[(2S,5R,6R)-5-azido-6-[(1R,2R,3S,4S,5R,6S)-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyrrolidone]-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyrrolidone] =Toluene-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-4-[[(2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoyl]amino]-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy-cyclohexyloxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (2.8 g, 1.99 mmol) and Pd(OH)2 (20% wet, 8.4 g, 12 mmol). The mixture was degassed by 3x vacuum-H2 cycles followed by bubbling with hydrogen (1 atm) for 2 h followed by bubbling with N2 for 15 min. The suspension was filtered on Celite followed by washing with MeOH (200 mL), water (200 mL). The volatiles were evaporated under reduced pressure (rotavap bath temperature <35°C). Use 10mM NH4HCO35-15% MeCN (pH 10) by preparation HPLC by material purification, then freeze-drying, obtain solid.Solid is dissolved in water (50mL), then ammonium sulfate (3 equivalents, 790mg, 5.98mmol) is added.The mixture is stirred at room temperature for 5 minutes, then filtered with 0.40uM syringe filter, and added dropwise to MeOH (1.5L) under vigorous stirring.The suspension is filtered on fine frit (fine frit), and rinsed with MeOH (50mL).The mother liquor is discarded, and the solid is dissolved in water (40mL), freeze-dried, so as to obtain the title compound (1100mg, 54%) in solid.Compound 3 (1.1g, 53%) is white solid. 1H NMR (400 MHz, D2O) δ 5.87 (d, J = 3.6 Hz, 1H), 5.29 (s, 1H), 5.25-5.05 (m, 2H), 4.47-4.17 (m, 4H), 4.18-3.97 (m, 3H), 3.88-3.78 (m, 3H), 3.77-3.68 (m, 3H), 3.62 (dd, J = 12.5, 5.6 Hz, 1H), 3.54 (t, J = 9.9 Hz, 1H), 3.49-3.44 (m, 1H), 3.44-3.16 (m, 7H), 1.99-1.82 (m, 3H), 1.49-1.34 (m, 1H), 1.19 (d, J = 6.7 Hz, 3H). Note: 20 exchangeable protons were not observed.

[0874] Example 9. Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R, 6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4- (((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy 5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclopentane Hexyl)-3-fluoro-2-hydroxybutyramide (Compound 4)

[0875]

[0876] Step A: Synthesis of (3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl(E)-2,2,2-trifluoro-N-phenylacetimidate

[0877]

[0878] To a mixture of N-[(1R)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (Ring A3, Example 3; 1.30 g, 2.85 mmol) and Cs2CO3 (1.86 g, 5.70 mmol) in DCM (10.0 mL) was added 2,2,2-trifluoro-N-phenylacetimidyl chloride (1.19 g, 5.70 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was filtered, the solid was rinsed with DCM, and the solvent was concentrated under reduced pressure. The residue was purified on silica gel (dry loading) using a gradient of 0 to 35% EtOAc in hexanes to give the title compound (1.20 g, 73%) as an oil. MS: no ionization.

[0879] Step B: Synthesis of ((1R,2R,3S,4R,5R,6S)-4-(((3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-2,5,6-trihydroxycyclohexane-1,3-diyl)dicarboxaminic acid dibenzyl ester

[0880]

[0881] [[[(2R, 3R, 6S)-3-azido-6-[(1R)-1-[benzyl(benzyloxycarbonyl)amino]ethyl]tetrahydropyran-2-yl](1E)-2,2,2-trifluoro-N-phenyl-acetimidate (2.47 g, 4.25 mmol) and [(1S, 2R, 3S, 4S, 5R, 6R)-3,4-diacetoxy-2,6-bis(benzyloxycarbonylamino)-5-hydroxy-cyclohexyl]acetate (ring B1, 3.28 g, 5.73 mmol) were co-evaporated with anhydrous toluene (2x3 mL). The mixture was dried under reduced pressure for 30 minutes. The residue was taken up in anhydrous DCM (100 mL) and then transferred via a syringe to a column filled with pre-activated and sieved RBF. Et2O (60mL) was added. The mixture was stirred at room temperature for 20 minutes and then cooled to 0°C. Bi(OTf)3 (3.07g, 4.67mmol) was added once, and the mixture was stirred at room temperature for 4h. A saturated aqueous solution (100mL) of NH4Cl was added. The mixture was filtered on diatomaceous earth. The aqueous layer was extracted with DCM respectively. The combined organic layer was dried (MgSO4), filtered, and concentrated under reduced pressure. The residue was purified on silica gel using a gradient of 0 to 100% EtOAc in hexane. The solid was dissolved in MeOH (100mL), and K2CO3 (1.52g, 11.0mmol) was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure (rotary evaporator water bath not exceeding 35°C). Water (100mL) and EtOAc (100mL) were added. The aqueous layer was extracted with EtOAc respectively. The combined organic layers were dried (MgSO4), filtered, and concentrated under reduced pressure to give the title compound (2.30 g, 99%) as a white solid. m / z (ESI) [M+H] + ;839.3.

[0882] Step C: Synthesis of ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarboxaminic acid dibenzyl ester

[0883]

[0884] A solution of N-[(1R)-1-[(2S, 5R, 6R)-5-azido-6-[(1R, 2S, 3R, 4R, 5S, 6R)-2,4-bis(benzyloxycarbonylamino)-3,5,6-trihydroxy-cyclohexyloxy]tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (2.30 g, 2.60 mmol), 1,1-dimethoxycyclohexane (2.38 mL, 15.6 mmol) and PPTS (262 mg, 1.01 mmol, dried over V10) in dry DME (111 mL) was stirred at 90 ° C for 30 minutes. The mixture was cooled to room temperature and concentrated under reduced pressure. The material was purified by ACCQ using 10 mM ammonium formate (pH 3.8) and MeCN. Subsequently, the product was diluted in EtOAc and washed with brine. The organic layer was dried (MgSO4), filtered, and concentrated under reduced pressure to afford (1.40 g, 59%). m / z (ESI) [M+H] + ;919.4.

[0885] Step D: Synthesis of ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarboxaminic acid dibenzyl ester

[0886]

[0887] To a mixture of ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxyhexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarboxaminic acid dibenzyl ester (1.40 g, 1.52 mmol) and BnBr (2.03 mL, 17.1 mmol) in THF (75 mL) at 0°C under nitrogen was added sodium hydride (60% dispersion in mineral oil, 141 mg, 3.51 mmol). The mixture was stirred at 0°C for 4 h, then AcOH (0.24 mL, 4.13 mL) was added. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (120 g column) using a gradient of 0 to 60% EtOAc in hexanes to give the title compound as a solid (1.43 g, 93%). m / z (ESI) [M+H] + ;1009.5.

[0888] Step E: Synthesis of dibenzyl ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarbamate

[0889]

[0890] A mixture of ((3aS,4R,5S,6S,7R,7aS)-4-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-6-(benzyloxy)hexahydrospiro[benzo[d][1,3]dioxolane-2,1'-cyclohexane]-5,7-diyl)dicarboxamate dibenzyl ester (1.44 g, 1.43 mmol) in AcOH (18 mL), water (8.0 mL) and THF (27 mL) was heated at 80 °C for 24 h. The mixture was cooled to room temperature and the THF was removed under reduced pressure. Water (100 mL) was added and a suspension was observed. The suspension was filtered over a medium frit. The solid was rinsed with water and dried under reduced pressure to obtain the title compound (1.21 g, 91%). m / z: ES+[M+H] + 929.6.

[0891] Step F: Synthesis of benzyl ((R)-1-((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydroxybenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (ring AB3)

[0892]

[0893] To a solution of ((1S,2R,3S,4R,5R,6S)-4-(((2R,3R,6S)-3-azido-6-((benzyl((benzyloxy)carbonyl)amino)methyl)tetrahydro-2H-pyran-2-yl)oxy)-2-(benzyloxy)-5,6-dihydroxycyclohexane-1,3-diyl)dicarboxamate (1.22 g, 1.31 mmol) in dry DMF (20 mL) at 0°C was added sodium hydride (210 mg, 5.25 mmol, 60% oil dispersion). The mixture was stirred at 0°C for 3 h, then AcOH (0.64 mL, 11.2 mmol) was added. The solution was poured into water (200 mL). The suspension was filtered and the solid was rinsed with water (20 mL) and dried under reduced pressure to give the title compound (830 mg, 77%) as a white solid. m / z(ESI)[M+H] + ;821.3.

[0894] Step G: Synthesis of benzyl ((1R)-1-((2S,5R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate

[0895]

[0896] To a mixture of [(2R, 3R, 4R)-4-acetoxy-3-[(2R, 3R, 4R, 5R, 6S)-4, 5-diacetoxy-3-azido-6-(azidomethyl)tetrahydropyran-2-yl]oxy-5-hydroxy-tetrahydrofuran-2-yl]methyl acetate (ring CD1, International Patent Application Publication No. WO2019194858, incorporated by reference, 961 mg, 1.81 mmol) and K2CO3 (429 mg, 3.11 mmol) in dry DCM (8 mL) was added CCl3CN (0.31 mL, 3.11 mmol). The mixture was stirred at room temperature for 18 h, then filtered on celite, rinsed with DCM, and concentrated under reduced pressure. To the previous crude material was added benzyl ((R)-1-((2S,5R,6R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (ring AB3; 425 mg, 0.518 mmol) in dry DCM (8 mL) and then transferred to a RBF packed with pre-activated 3A and 4A sieves. The mixture was cooled to -78 °C and then BF3-OEt2 (0.96 mL, 7.81 mmol) was added dropwise at -78 °C. The dry ice bath was removed once the addition was complete. The mixture was stirred at room temperature for 1h, diluted with a saturated aqueous solution of NaHCO3 (10mL), filtered through diatomaceous earth, and the filter cake was washed with DCM (20mL). The separated aqueous layer was extracted with DCM (3x5mL) respectively. The combined organic layer was washed with brine (15mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The residue was incorporated into 1,4-dioxane (4.0mL) and water (4.0mL), followed by the addition of LiOH-H2O (652mg, 15.5mmol), and the mixture was stirred at 40°C for 18h. The mixture was cooled to room temperature, and then 1,4-dioxane was removed under reduced pressure, followed by the addition of EtOAc (10mL). The separated aqueous layer was extracted with EtOAc (3x10mL) respectively. The combined organic layer was dried (MgSO4), filtered, and concentrated under reduced pressure. The residue was purified by preparative ACCQ using a gradient of MeCN in 10 mM ammonium bicarbonate (pH 10) to give the title compound as a solid (209 mg, 35% after 2 steps). m / z (ESI) [M+H] + ;1139.5.

[0897] Step H: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butyramide)-6-(((2R,3R ,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0898]

[0899] To a solution of benzyl ((1R)-1-((2S,5R)-5-azido-6-(((3aR,4S,5S,6R,7S,7aS)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-7-hydroxy-2-oxooctahydrobenzo[d]oxazol-6-yl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)(benzyl)carbamate (104 mg, 0.091 mmol) and (2R,3R)-4-azido-3-fluoro-2-[(4-methoxyphenyl)methoxy]butanoic acid (F-HABA, Example 5; 44 mg, 0.157 mmol) in anhydrous THF (5 mL) was added DMTMM chloride (44 mg, 0.160 mmol). The mixture was stirred at room temperature for 18 h then diluted with saturated aqueous NH4Cl (3 mL) and EtOAc (5 mL). The separated aqueous layer was extracted with EtOAc (3x4mL) respectively. The combined organic layer was washed with brine (5mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by preparative HPLC (ACCQ, BEH C18 column) using MeCN of ammonium formate (10mM, pH 3.8) and lyophilized to give the title compound (103mg, 80%) as a white solid. m / z(ESI)[M+H] + :M+H+:1404.6.

[0900] Step I: Synthesis of ((2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-3-fluoro-2-hydroxybutanamide (Compound 4)

[0901]

[0902] To a suspension of Pd(OH)2 (359 mg, 0.51 mmol, 20% wet) in a mixture of AcOH (7.50 mL) and water (3.0 mL) was added 2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2S,3S,4R,5S,6R)-3-((2R,3R)-4-azido-3-fluoro-2-((4-methoxybenzyl)oxy)butyramido)-6-(((2R,3R,6S A solution of 5-(((benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-(((benzyloxy)carbonyl)amino)-2-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (104 mg, 0.074 mmol) in MeOH (9.0 mL) was degassed by 3x vacuum-H2 cycles followed by bubbling with hydrogen for 10 min. The mixture was stirred for 2.5 hours under a hydrogen atmosphere (1 atm), degassed with N for 5 minutes, then filtered on a 0.22 μM PVDF filter syringe, washed with MeOH (3x3 mL) and water (2x2 mL), and then evaporated under reduced pressure (rotary evaporation bath temperature <35 ° C). 10 mM NH was used 5% MeCN (pH 10) in HCO3 to purify the material by filtration, and lyophilized to obtain a solid. At 0 ° C, a solution of the above solid in H2O (3 mL) was dropwise added with H2SO4 (0.05 M in H2O), until pH 4, then lyophilized to obtain a solid, which was dissolved in water (2 mL), followed by MeOH (10 mL) to obtain a precipitate. The precipitate was filtered through a 0.22 μM PVDF syringe filter, rinsed with MeOH (2x5 mL), and then washed with water (5 mL). The solid was dissolved in water (10 mL) and lyophilized to give the title compound 4 (30 mg, 40%) as a white solid. 1 H NMR(500MHz,D2O)δ5.95-5.88(m,1H),5.47-5.24(m,3H),4.57(d,J=10.2Hz ,1H),4.51-4.34(m,3H),4.27(d,J=3.7Hz,2H),4.18(d,J=12.1Hz,1H),4.04 -3.88(m,5H),3.82-3.59(m,5H),3.58-3.37(m,5H),3.33-3.22(m,2H),2.1 2-2.00(m,2H),1.96-1.89(m,1H),1.69-1.58(m,1H),1.31(d,J=6.9Hz,3H).

[0903] Example 10. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)- 3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R, 3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (Hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutyramide (Compound 5)

[0904]

[0905] Step A: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0906]

[0907] At 0°C, N-[(1R)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)- To a solution of [3-hydroxy-cyclohexyloxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (90 mg, 0.079 mmol), (2S)-2-benzyloxy-4-(benzyloxycarbonylamino)butyric acid (HABA; 40.7 mg, 0.119 mmol) and DIPEA (0.04 mL, 0.237 mmol) in anhydrous DMF (3 mL) was added PyBOP (49.3 mg, 0.0948 mol). The mixture was warmed to room temperature and stirred for 18 h. The mixture was poured into water (5 mL). 1N HCl was used to adjust the pH to 4, followed by addition of EtOAc (40 mL). The separated aqueous layers were extracted with EtOAc (5 mL x3), respectively. The combined organic layers were washed with brine (5 mL), dried (MgSO ), filtered, and concentrated under reduced pressure. The material was purified by reverse phase column (ACCQ) using 60-80% ACN gradient in ammonium formate (10mM, pH 3.8) and concentrated under reduced pressure. The material was then dissolved in EtOAc (30mL) and water (5mL). The separated aqueous layer was extracted with EtOAc (3x5mL) respectively. The combined organic layer was washed with brine (5mL), dried (MgSO4), filtered, and concentrated under reduced pressure to give the title compound (79.3mg, 69%) as a white solid. m / z(ESI)[M+H] + :1464.5, HPLC A(05)tR=2.65min.

[0908] Step B: Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 5)

[0909]

[0910] To a suspension of Pd(OH)2 (20% wet, 261 mg, 0.372 mmol) in a mixture of AcOH (5.5 mL) and water (2.2 mL) was added (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((R)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro- To a solution of ((((benzyloxy)carbonyl)amino)butyramido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (79 mg, 0.0539 mmol) in MeOH (6.5 mL) was degassed by 3x vacuum-H cycles followed by bubbling with hydrogen for 10 min. The mixture was stirred under hydrogen bubbling (1 atm, refilling the balloon with H as necessary) for 4 h. The suspension is degassed with N2 for 15 minutes, then filtered on a 0.22 μM PVDF filter, washed with MeOH (3x5mL) and water (2x5mL), and volatiles are evaporated under reduced pressure (rotary evaporation bath temperature <35 ° C). Use 10mM NH4HCO3 in 5% MeCN (pH 10) to purify the material by filtration, and freeze-dry to obtain a solid. At 0 ° C, to the above-mentioned solid in H2O (3mL) solution dropwise add H2SO4 (0.05M in H2O), until pH 4, then freeze-dried. The solid is dissolved in water (2mL), followed by MeOH (10mL). The suspension is filtered through a 0.22uM PVDF syringe filter, rinsed with MeOH (2x5mL), then washed with water (5mL). The solid is dissolved in water (10mL) and freeze-dried to obtain the title compound 5 (26mg, 48%) as a solid. 1H NMR(400MHz,D2O)δ5.76(dd,J=10.0,3.7Hz,1H),5.27(dd,J=6.2,2.8Hz,1H),5.16(d,J=1.8Hz,1 H),4.40(dd,J=7.2,4.7Hz,1H),4.35-4.28(m,1H),4.26-4.18(m,2H),4.10(t,J=3.3Hz,2H),4.03 -3.95(m,1H),3.93-3.67(m,5H),3.64-3.19(m,8H),3.18-3.08(m,1H),3.08-2.98(m,2H),2.13-1 .99(m,1H),1.98-1.84(m,3H),1.81-1.70(m,1H),1.55-1.40(m,1H),1.14(dd,J=6.9,1.2Hz,3H). m / z(ESI)[M+H] + ;714.4.

[0911] Example 11. Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)- 3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R, 3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5- (Hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutyramide (Compound 6)

[0912]

[0913] Step A: Synthesis of (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butanamido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate

[0914]

[0915] At 0°C, N-[(1S)-1-[(2S,5R,6R)-6-[(1R,2R,3S,4S,5R,6S)-4-amino-2-[(2S,3R,4S,5R)-4-[(2R,3R,4R,5S,6S)-3-azido-6-(azidomethyl)-4,5-dihydroxy-tetrahydropyran-2-yl]oxy-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]oxy-5-benzyloxy-6-(benzyloxycarbonylamino)-3-hydroxy- To a solution of [(2S)-2-benzyloxy]-5-azido-tetrahydropyran-2-yl]ethyl]-N-benzyl-carbamic acid benzyl ester (Example 8, Step J; 133 mg, 0.117 mmol), (2S)-2-benzyloxy-4-(benzyloxycarbonylamino)butanoic acid (HABA; 60.1 mg, 0.175 mmol) and DIPEA (0.061 mL, 0.350 mmol) in anhydrous DMF (4.0 mL) was added PyBOP (73.0 mg, 0.140 mol). The mixture was warmed to room temperature and stirred for 18 h. The mixture was poured into water (5.0 mL) and EtOAc (5.0 mL) was subsequently added. The separated aqueous layers were each extracted with EtOAc (3 x 5.0 mL). The combined organic layers were washed with brine (5.0 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The material was purified by reverse phase column (C18, 40 g) using a 0-100% ACN gradient of ammonium formate (10 mM, pH 3.8) and lyophilized to give the title compound as a white solid (66 mg, 38%). m / z (ESI) [M+H] + :1464.5, HPLC A(05)tR=2.94min.

[0916] Step B: Synthesis of (S)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R,6S)-3-amino-6-((S)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)-4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclohexyl)-2-hydroxybutanamide (Compound 6)

[0917]

[0918] To a suspension of Pd(OH)2 (20% wet, 218 mg, 0.311 mmol) in a mixture of AcOH (5.0 mL) and water (3.0 mL) was added (2S,3R,4R,5R,6R)-6-(((2R,3R,4R,5S)-4-acetoxy-2-(acetoxymethyl)-5-(((1R,2R,3S,4R,5S,6S)-2-(((2R,3R,6S)-3-azido-6-((S)-1-(benzyl((benzyloxy)methyl)- To this was added 4-((((benzyloxy)carbonyl)amino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-4-(benzyloxy)-5-((S)-2-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)butyramido)-3-(((benzyloxy)carbonyl)amino)-6-hydroxycyclohexyl)oxy)tetrahydrofuran-3-yl)oxy)-5-azido-2-(azidomethyl)tetrahydro-2H-pyran-3,4-diyl diacetate (66 mg, 0.0451 mmol) in MeOH (7 mL). Hydrogen was bubbled through for 3 h, followed by degassing with N2 for 15 min. The suspension was filtered on a 0.22 μM PVDF filter, washed with MeOH (8.0 mL), water (8.0 mL), and DCM (8.0 mL), and the volatiles evaporated under reduced pressure (rotary evaporator; bath temperature <35 °C). Use 10mM NH4HCO3 in 5% MeCN (pH 10) by filtration crude material purification and lyophilization to obtain solid. At 0 ° C to the above solid in H2O (3mL) solution dropwise add H2SO4 (0.05M in H2O), until pH 4, then lyophilized. The solid is dissolved in water (2mL), followed by MeOH (10mL). The suspension is filtered through a 0.22uM PVDF syringe filter, rinsed with MeOH (2x5mL), then washed with water (5mL). The solid is dissolved in water (10mL) and lyophilized to obtain the title product compound 6 (30mg, 66%) as a solid. 1 H NMR (500MHz, D2O) δ6.04 (d, J = 3 Hz, 1H), 5.47 (m, 1H), 5.33 (m, 2H), 4.58-4.55 (m, 1H), 4.48-4.34 (m, 3H), 4.30-4.21 (m, 3H), 4.05-3.85 (m, 6H), 3.81-3.66 (m, 2H), 3.63 (m, 1H), 3.61-3.14 (m, 7H), 2.26-2.16 (m, 1H), 2.11-2.01 (m, 4H), 1.63-1.50 (m, 1H), 1.35 (d, J = 6.0 Hz, 3H). Note: 20 exchangeable protons were not observed. m / z (ESI) [M+H] + :714.3,HPLC A(05)tR=0.33min.

[0919] Example 12: Synthesis of (2R,3R)-4-amino-N-((1R,2S,3S,4R,5R,6S)-3-amino-4-(((2R,3R, 6S)-3-amino-6-((S)-1-(methylamino)ethyl)tetrahydro-2H-pyran-2-yl)oxy)-5-(((2S,3R,4S,5R)- 4-(((2R,3R,4R,5S,6S)-3-amino-6-(aminomethyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-3- Hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)oxy)-2,6-dihydroxycyclic Hexyl)-3-fluoro-2-hydroxybutyramide (Compound 7)

[0920]

[0921] Step A: Synthesis of (S)-N-[(1S)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfenamide

[0922]

[0923] To a mixture of (S)-N-[(1S)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-2-methyl-propane-2-sulfinamide (83.0 g, 334 mmol) in THF (1.0 L) at 0°C under nitrogen was added sodium hydride (60% in mineral oil, 19.2 g, 500 mmol). The mixture was stirred at 0°C for 15 minutes. Iodomethane (41.5 mL, 667 mmol) was added and the mixture was stirred at room temperature for 3 hours. The mixture was diluted with saturated aqueous NH4Cl (500.0 mL), water (200.0 mL), and the aqueous phase was extracted with ethyl acetate (3x250 mL). The combined organic phases were washed with brine (250 mL), dried (MgSO4), filtered, and concentrated under reduced pressure. The crude residue (94.3 g) was split into two portions, each portion was purified by silica gel chromatography (350 g, Biotage HC, liquid load) using a gradient of 0 to 30% EtOAc in hexanes to give the title compound, and mixed portions. The mixed portions were subjected to a third round of purification, these portions were purified by silica gel chromatography (350 g HC, liquid load) using a gradient of 0 to 20% EtOAc in hexanes. All pure portions from these three purifications were combined to give the title compound (30.2 g, 31%, -85% purity). m / z: ES+[M+H]+246.2; HPLC(A05); tR=2.29m. (desired title product). m / z: ES+[M+H]+246.2; HPLC(A05); tR=2.34 min. (undesired diastereoisomer).

[0924] Step B: Synthesis of (S)-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfenamide

[0925]

[0926] At 0 ° C, to a suspension of (S)-N-[(1S)-1-[(2S)-3,4-dihydro-2H-pyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfinamide (30.4 g, 124 mmol) and NaHCO (31.2 g, 372 mmol) in ACN (500.0 mL) and H2O (500.0 mL) was added iodine (31.4 g, 124 mmol) portionwise. The mixture was stirred at 0 ° C for 30 minutes. The mixture was stirred at room temperature for 2 h30. The mixture was diluted with a saturated aqueous solution of Na2S2O3 (500.0 mL) and the aqueous layer was extracted with EtOAc (3x500.0 mL). The combined organic phases were washed with brine (300.0 mL), dried (MgSO4), filtered and concentrated to give the title compound (47.7 g, 99% crude) as an oil. The compound was used without further purification. m / z(ESI)[M+Na] + : 412.0, HPLC tR = 2.29 min and 2.40 min (desired product and diastereomers).

[0927] Step C: Synthesis of benzyl-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate

[0928]

[0929] To a solution of (S)-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N,2-dimethyl-propane-2-sulfinamide (47.7 g, 123 mmol) in dioxane (750.0 mL) was added 1 M aqueous HCl (147.0 mL, 147 mmol) at 5-10°C. The mixture was stirred at 5-10°C for 45 minutes, followed by the addition of PBS 10x buffer (~2.0 L) until the pH reached ~6. The mixture was warmed to room temperature, followed by the addition of CbzCl (35.0 mL, 245 mmol). The mixture was stirred at room temperature for 3 hours, followed by the addition of EtOAc (500.0 mL). The separated aqueous layers were extracted with EtOAc (3x400.0 mL), respectively. The combined organic layers were washed with brine, dried (MgSO4), filtered, and concentrated under reduced pressure. The residue was divided into two portions (2 x 45 g crude) and subsequently purified on silica (330 g, liquid load) using EtOAc and hexanes (0 to 50%) to give the title compound (15.0 g, 27%) as an oil. m / z (ESI) [M+Na]+: 442.0, HPLC, A(05) tR=2.49 min, 2.54 min. (title product).

[0930] Step D: Synthesis of benzyl-N-[(1S)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate

[0931]

[0932] Oxidation:

[0933] Benzyl-N-[(1S)-1-[(2S)-6-hydroxy-5-iodo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate (15.0 g, 35.8 mmol) and To a mixture of sieves (5.0 g) in DCM (250.0 mL) was added PDC (33.6 g, 89.4 mmol). The mixture was stirred at room temperature for 18 hours, then filtered on a pad of silica gel, rinsed with EtOAc (3 x 350.0 mL), and concentrated under reduced pressure. m / z (ESI) [M+H] + : 418.0, HPLC (A05); tR = 2.44 min (oxidized intermediate).

[0934] Azide:

[0935] The residue was taken up in DMF (150.0 mL) and cooled to 0 ° C. NaN3 (2.21 g, 34.0 mmol) was added and the mixture was subsequently stirred at 0 ° C for 1 h. Brine (500.0 mL) and Et2O (150.0 mL) were added. The aqueous layer was extracted with Et2O (2x150.0 mL) respectively. The combined organic layers were dried (MgSO4) and concentrated under reduced pressure. The material was purified by MPLC on silica (220 g, Biotage HC) using a gradient of 0-60% EtOAc in hexane to give the title compound (8.0 g, 67% after 2 steps). m / z(ESI)[M+K] + : 371.4, HPLC (A05); tR = 2.33 min (title compound).

[0936] Step E: Synthesis of benzyl-N-[(1S)-1-[(2S,5R)-5-azido-6-hydroxy-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate

[0937]

[0938] To a solution of benzyl-N-[(1S)-1-[(2S)-5-azido-6-oxo-tetrahydropyran-2-yl]ethyl]-N-methyl-carbamate (8.0 g, 24.1 mmol) in DCM (250.0 mL) was added DIBAL-H (1 M in toluene, 43.3 mL, 43.3 mmol) at -78 °C...

Claims

1. A compound having a structure according to formula (III): or a pharmaceutically acceptable salt or solvate thereof, in: R a is H or methyl; and R b H, C 1-6 Alkyl, or having a structure according to formula (II): in: Q is NH, O or S; n is an integer from 0 to 4; R 1 is hydrogen or C 1-3 alkyl; R 2 and R 3 Each occurrence is independently selected from: hydrogen, alkyl, halogen, and -OH; and R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b are each independently selected from hydrogen and C 1-3 Alkyl; or R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

2. The compound of claim 1, wherein R a For H.

3. The compound as claimed in claim 1, wherein R a It is methyl.

4. The compound according to any one of claims 1 to 3, wherein the compound of formula (III) is selected from:

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

6. The compound of claim 5, wherein the compound of formula (III) has a structure selected from the following:

7. The compound according to any one of claims 1 to 3, wherein R b Selected from:

8. A compound as described in any one of claims 1 to 3, wherein R b Selected from:

9. The compound of claim 8, wherein the compound of formula (III) has a structure selected from the group consisting of:

10. A compound having a structure according to formula (I): or a pharmaceutically acceptable salt or solvate thereof, in: R a is hydrogen (H) or methyl; and R b C 1-6 Alkyl, or having a structure according to formula (II): in: Q is NH, O or S; n is an integer from 0 to 4; R 1 is hydrogen or C 1-3 alkyl; R 2 and R 3 Each occurrence is independently selected from: hydrogen, alkyl, halogen, and -OH; and R 4 H, C 1-3 Alkyl, or -C(=NH)NR 4a R 4b , where R 4a and R 4b Each independently selected from H and C 1-3 Alkyl; or R 1 and R 4 Together with the atoms to which they are attached they form a heterocycloalkyl ring system containing at least one N.

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

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

13. The compound according to any one of claims 10 to 12, wherein R b Selected from:

14. The compound according to any one of claims 10 to 13, wherein R b Selected from:

15. The compound of claim 10, wherein the compound of formula (I) has a structure selected from the group consisting of:

16. A compound as described in any one of claims 10-12, wherein R b It is ethyl.

17. The compound of claim 16, wherein the compound of formula (I) has the following structure:

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

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

20. The method of claim 19, wherein the bacterial infection is caused by Gram-positive bacteria, Gram-negative bacteria, aerobic bacteria, or facultative anaerobic bacteria.

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

22. The method of claim 20, wherein the bacterial infection is an infection caused by Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter krusei, Proteus mirabilis, Bacillus anthracis, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus vulgaris, Francisella tularensis, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei, or Yersinia pestis.

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

24. The method of claim 23, wherein the Gram-negative bacterial infection is an infection caused by Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Burkholderia cepacia, or Alcaligenes xylosoxidans.

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

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

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

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

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