Glycomimetic Ligands

Abstract

Glycan compounds and particles comprising glycan compounds are described herein. The compounds and particles described herein are useful in methods for treating immune and inflammatory related diseases, or pharma- ceutically acceptable salts thereof.

Description

[Technical Field]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 295,698, filed December 31, 2021. The entire teachings of the above application are incorporated herein by reference. [Background technology]

[0002] There is a need for improved compositions and methods for modulating sialic acid-binding self-associated pattern recognition receptors known as sialic acid-binding immunoglobulin-type lectins (Siglecs) for the treatment of diseases caused by acute, chronic, or aberrant immune system activation. Summary of the Invention [Means for solving the problem]

[0003] In an example embodiment, the present invention provides a compound having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof.

[0004] In formulae (S-1), (S-2), (D-1) and (D-2), R1 is, independently at each occurrence, -C(O)-A, where A is C1-C6 alkyl, C6-C 18 Aryl, (C6-C 18 )aryl(C1-C3)alkyl, 5- to 18-membered heteroaryl, (5- to 18-membered)heteroaryl(C1-C3)alkyl, C3-C8 cycloalkyl, (C3-C8)cycloalkyl(C1-C3)alkyl, 5- to 8-membered heterocycloalkyl, or (5- to 8-membered)heterocycloalkyl(C1-C3)alkyl, wherein one or two carbon atoms within the alkyl portion of A are each independently optionally replaced with a heteroatom selected from N, O, or S, and A is selected from 1 to 3 R 11 groups, and each of the R 11The group is C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, halogen, C6-C 12 aryl, 5- to 12-membered heteroaryl, cyano, or two groups R 11 R together with the atoms to which they are attached form a 5-7 membered heterocyclyl having 1-3 heteroatoms selected from N, O or S. 11 are each independently optionally substituted with 1 to 3 substituents selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or cyano; and R is independently selected at each occurrence from -R L -R F and R L is independently expressed as -O-(C1-C 12 ) Alkylenyl-, -O-, -S-, -NR 100 -, -S-(C1~C 12 ) Alkylenyl-, -NR 101 -(C1~C 12 ) Alkylenyl-, -NR 101 aO-(C1~C 12 )Alkylenyl-;-O-(CH2CH2O) m -, -O-(CH2CH2O) k -(CH2CH2)-, -NR 102 -X 100 -(C1~C 12 ) Alkylenyl-, -NR 102a -NR 102b -C(O)-(C1~C 12 ) alkylenyl, and R 100 , R 101 , R 101a , R 102 , R 102a and R 102b are each independently H or C1-C3 alkyl, and X 100 is —O— or —NH—, m and k each independently represent an integer of 1 to 12; R Frepresents independently at each occurrence H, C1-C3 alkyl, -NH2, -NH-Fmoc, -NH-Boc, -NH-CBz, -NH-Troc, -NH-TFA, mono(C1-C3)alkylamino, di(C1-C3)alkylamino; -C(O)-R 103 (In the formula, R 103 is -H, -OH, or (C1-C3) alkyl); [ka] or a click chemistry reagent.

[0005] In another exemplary embodiment, the present invention provides a compound of formula (2): GLP (2) The particle includes a compound represented by the formula:

[0006] In structural formula (2), P is a biocompatible polymer. For example, the biocompatible polymer may include at least one of polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), poly(ethylene glycol), polyethylene oxide, poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (Pluronic F127), polyoxyethylene-polyoxypropylene block copolymer (Pluronic F68), poloxamer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan; L is a covalent linker; and G is a polymer having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof. Values ​​and example values ​​of the variable R1 are as defined above in connection with formulae (S-1), (S-2), (D-1) and (D-2).

[0007] In another exemplary embodiment, the present invention provides a compound of formula (2): GLP (2) wherein P is a biocompatible polymer. For example, the biocompatible polymer can include at least one of polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), poly(ethylene glycol), polyethylene oxide, poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (Pluronic F127), polyoxyethylene-polyoxypropylene block copolymer (Pluronic F68), poloxamer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan; L is a covalent linker; and G is any one of the moieties represented by Structural Formulas (S-1A), (S-2A), (D-1A), and (D-2A), or a pharmaceutically acceptable salt thereof. A method of making particles comprising a compound represented by the symbol [ka] represents the point of attachment to L, and values ​​and example values ​​of the variable R1 are as defined above in relation to formulae (S-1), (S-2), (D-1) and (D-2). GR F1 (I) The compound represented by structural formula (II) PR F2 (II) (In the formula, R F1 and R F2 are reactive moieties, respectively) and a compound represented by the moiety R F1 and R F2 with each other under conditions suitable to thereby produce a covalently linked -L- moiety.

[0008] In various example embodiments, the compounds and particles described herein are useful in methods for treating diseases and disorders that respond to modulation (e.g., amplification, reduction, or elimination) of Siglec receptor activity. Such diseases and disorders include, but are not limited to, cancer, immune-related, and inflammation-related diseases and disorders. For example, the compounds and particles described herein are useful in methods for treating a disorder selected from cancer, ophthalmic disease, fibrosis, parasitic inflammation, fungal inflammation, viral inflammation, autoimmune inflammation, neurogenic inflammation, neurodegeneration, skin inflammation, renal inflammation, cardiovascular disease, gastrointestinal inflammation, or rheumatic disease.

[0009] In other embodiments, the compounds and particles described herein are useful for the treatment of a disorder selected from cancer (e.g., breast cancer, non-small cell lung cancer (NSCLC), prostate cancer, colorectal cancer, melanoma, pancreatic cancer, and myelofibrosis), diabetic retinopathy, idiopathic pulmonary fibrosis, hepatic fibrosis, sickle cell anemia, and acute respiratory distress syndrome (ARDS).

[0010] In a further exemplary embodiment, the present invention provides a compound of the following structural formula: [ka] or a pharmaceutically acceptable salt thereof. In formula (S000-C), R is independently at each occurrence -R L -R F and R L are, independently for each occurrence, -O-, -S-, and -NR 100 -, -O-(C1~C 12 ) Alkylenyl-, -S-(C1-C 12 ) Alkylenyl-, -NR 101 -(C1~C 12 ) Alkylenyl-, -NR 101a -O-(C1~C 12 )Alkylenyl-;-O-(CH2CH2O) m -, -O-(CH2CH2O) k -(CH2CH2)-, -NR 102 -X 100 -(C1~C 12 ) Alkylenyl-, -NR 102a -NR102b -C(O)-(C1~C 12 ) alkylenyl, and R 100 , R 101 , R 101a , R 102 , R 102a and R 102b are each independently H or C1-C3 alkyl, and X 100 is —O— or —NH—, m and k each independently represent an integer of 1 to 12; R F represents independently at each occurrence H, C1-C3 alkyl, -NH2, -NH-Fmoc, -NH-Boc, -NH-CBz, -NH-Troc, -NH-TFA, mono(C1-C3)alkylamino, di(C1-C3 alkyl)amino; -C(O)-R 103 (In the formula, R 103 is -H, -OH, or (C1-C3) alkyl); [ka] or a click chemistry reagent, which is a moiety represented by -R L -R F is not -OH.

[0011] In another embodiment, the present invention provides a compound of the following structural formula: GLP wherein P is a biocompatible polymer. For example, the biocompatible polymer can include at least one of polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), poly(ethylene glycol), polyethylene oxide, poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (Pluronic F127), polyoxyethylene-polyoxypropylene block copolymer (Pluronic F68), poloxamer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan; L is a covalent linker; and G is a polymer having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof) are particles containing a compound represented by the symbol [ka] represents the point of attachment to L.

[0012] In another embodiment, the present invention is a method of treating a disorder in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of a compound of formula (S000-C) or a pharmaceutically acceptable salt thereof, or a compound of the following structural formula: GLP or a pharmaceutically acceptable salt thereof, wherein G is represented by the structural formula (S000-P), and the disorder is influenza.

[0013] In another exemplary embodiment, the present invention is any of the compounds listed in Table 4 or a pharmaceutically acceptable salt thereof.

[0014] In another exemplary embodiment, the present invention is a compound represented by any of the structural formulas listed in Figures 6B-6F, or a pharmaceutically acceptable salt thereof.

[0015] In another embodiment, the invention is a compound or particle described herein for use in therapy, e.g., for use in the treatment of diseases and disorders that respond to modulation (e.g., amplification, reduction, or elimination) of Siglec receptor activity. Such diseases and disorders include, but are not limited to, cancer, immune-related, and inflammatory-related diseases and disorders, such as those described herein.

[0016] In another embodiment, the invention is a compound or particle as described herein for the manufacture of a medicament for use in the treatment of diseases and disorders responsive to modulation (e.g., amplification, reduction, or elimination) of Siglec receptor activity. Such diseases and disorders include, but are not limited to, cancer, immune-related, and inflammatory-related diseases and disorders such as those described herein.

[0017] The compounds, particles, compositions and methods described herein can be used for the treatment of the disorders described herein and provide additional treatments for many difficult to treat diseases and disorders.

[0018] The foregoing will become apparent from the following more particular description of example embodiments of the invention, as illustrated in the accompanying drawings, in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating embodiments of the invention. [Brief explanation of the drawings]

[0019] [Figure 1A-B] 2D 1H-NMR spectra of compounds 22 and 23 of Scheme 1-6, which correspond to compounds 240 and 250 of Scheme 10-10. [Figure 2] 1 is a plot showing the results of tracking the reaction participants and products of Reaction Schemes 1-7 and Reaction Schemes 10-11 using hydrophilic interaction liquid chromatography / electrospray ionization mass spectrometry (HILIC-L / ESI-MS). [Figure 3A-M] 1 is a bar graph showing the results of microarray screening of compounds described herein for binding to the indicated Siglecs and viral particles (the vertical axis corresponds to fluorescence intensity measured in relative fluorescence units (RFU)). [Figure 4A-E] 1 is a sensorgram generated by biolayer interferometry (BLI) measurement of binding constants of certain compounds of the invention, as described in Example 6. [Figure 5] HSQC (600 MHz) 2D NMR spectrum of S106 azide. [Figure 6A-F] 1 shows the structural formula of the polymer-ligand conjugate. [Figure 7] 1 is a plot of THP-1 monocyte-derived macrophage viability in an MTT assay as a function of nanoparticle concentration. [Figure 8] 1 is a bar graph showing the suppression of TNF-α production in LPS-challenged THP-1 cells after incubation with nanoparticles described herein. [Figure 9] 1 is a bar graph showing the suppression of IL-6 production in LPS-challenged THP-1 cells after incubation with nanoparticles described herein. [Figure 10] 1 is a bar graph showing the suppression of VEGF production in LPS-challenged THP-1 cells after incubation with nanoparticles described herein. [Figure 11A-B] 1 is a bar graph showing the effect of tested nanoparticle formulations on fibrocyte differentiation. [Figure 12] 1 is a plot of ROS production in neutrophils 1 hour after PMA treatment as a function of nanoparticle dose. [Figures 13A-C] 1 is a plot of MPO (myeloperoxidase) levels in neutrophil supernatants after PMA treatment of neutrophils at different time points in the presence of the indicated nanoparticles. [Figure 14A-B] 1 is a plot showing quantification of LysoBrite-labeled BV-2 microglia using a fluorescence spectrophotometer after treatment with the indicated nanoparticle formulations. [Figure 15] FIG. 1 is a schematic diagram of a cell surface binding assay of recombinant Siglec-9 Fc. [Figure 16] Flow cytometry analysis of Siglec-9 Fc binding to PANC-1 cells + / - neuraminidase. [Figure 17] 1 shows flow cytometry analysis of Siglec-9 Fc binding to PANC-1 cells + / - PLGA-based nanoparticles. [Figure 18]1 shows the results of flow cytometry analysis of Siglec-9 Fc binding to PANC-1 cells + / - PLGA-based nanoparticles. [Figure 19] FIG. 1 is a schematic diagram of exemplary S-series ligands bound to FluoSpheres™ Neutravidin™-labeled microspheres. [Figure 20] 1 shows the results of flow cytometry analysis of Siglec-9 Fc binding to PANC-1 cells + / - FluoSpheres™ microspheres. [Figure 21] 1 shows a schematic diagram of the FLISA setup described herein. [Figure 22] 1 is a bar graph showing the results of a FLISA analysis of FluoSpheres™ microsphere binding to immobilized Siglec-9 Fc. [Figure 23] FIG. 1 is a schematic diagram of the binding assay setup using Siglec-9-expressing HEK293T cells. [Figure 24] 1 is a histogram showing the results of flow cytometry analysis of Siglec-9-PE binding to Siglec-9-expressing HEK293T cells + / - PLGA-based nanoparticles. DETAILED DESCRIPTION OF THE INVENTION

[0020] An example embodiment of the present invention is described below.

[0021] The present disclosure provides methods and compositions for modulating the activity of self-associated pattern recognition receptors, such as Siglecs (sialic acid-binding immunoglobulin-type lectins). The provided compositions include nanoparticles, microparticles, and other polymeric structures decorated with modified glycan structures that bind to, agonize, or antagonize self-associated molecular pattern recognition receptors and infectivity-associated sialic acid-binding moieties, enabling invasion, propagation, and evasion of immune surveillance in the host.

[0022] Binding to and / or stimulating or antagonizing the activity of such self-associated pattern recognition receptors can resolve innate, adaptive, multimodal, inflammatory, or complement-mediated immune responses, thereby providing treatment for diseases of (1) acute inflammation, such as viral, bacterial, allergen, transplant rejection, or autoimmune-induced inflammation; (2) chronic inflammation, such as chronic obstructive pulmonary disease or rheumatic diseases; and (3) congenital and adaptive chronic non-resolving inflammation, such as exudative or non-exudative macular degeneration or Alzheimer's disease.

[0023] The provided compositions can also be used to inhibit or antagonize self-associated molecular pattern recognition receptors that enable infectious agents, such as cancer cells, viral, bacterial, helminthic, parasitic, or damage-associated molecular patterns (DAMPs), to evade immune surveillance, detection, and clearance by the innate or adaptive immune system.

[0024] Stimulation of Siglecs 3, 5, 7, 8, 9, 10, 11, or 15 dephosphorylates all activated (phosphorylated) tyrosine kinases in a given cell, resulting in intracellular shutdown of activation in that particular cell. The modified oligosaccharide ligands and the presentation of these ligands to specific Siglec receptors determine their ability to stimulate, antagonize, or block receptor binding sites.

[0025] Antagonism of Siglec-14 or -16, which activates inflammation through immunoglobulin tyrosine kinase activation motifs (ITAMs), is another mechanism for inactivating inflammation. Stimulation of Siglec-14 or -16 activates the ITAMs, phosphorylating tyrosine residues within the ITAMs by SRC family kinases, resulting in a conformational change that allows the motifs to become docking sites for SH2 domain-containing proteins. Stimulation of Siglec-14 and -16 can be used to activate inflammation for the treatment of infectious diseases or in oncology settings.

[0026] Antagonism or blocking of the binding site of Siglec 3, 5, 7, 8, 9, 10, 11, or 15 is a method for treating conditions in which Siglecs are stimulated with self-associated molecular pattern (SAMP)-mimicking surface sialic acid ligands to avoid immune surveillance or activation. Conditions for which this method is used include cancer and infectious diseases. Cancers have been shown to evade immune activation of macrophages, natural killer (NK) cells, and monocytes by expressing sialic acid structures on their surface. Group B streptococci also express sialic acid ligands on their surface that bind to Siglec 7 to avoid immune invasion.

[0027] Siglec-9 has been extensively studied in both fibrocytes and fibrosis. Siglec-9 is one of the major Siglecs in human blood monocytes / macrophages and regulates innate immunity. Siglec-9 expression in alveolar and peripheral blood neutrophils was increased in patients with chronic obstructive pulmonary disease (COPD). These fibrocytes are unique cells that possess the proinflammatory properties of macrophages and the tissue remodeling properties of fibroblasts. Apart from fibrosis, neutrophils, including diabetic retinopathy models, have demonstrated a role for Siglec-9.

[0028] Glycophorin, a sialoglycoprotein present on the surface of red blood cells, inhibits NET release / neutrophil activation by binding sialic acid to Siglec-9 in the circulation. The interaction of lactoferrin with polySia increases the inhibition of NET release. Glycophorin A, the most abundant sialoglycoprotein on red blood cells, binds to neutrophil Siglec-9, a sialic acid-recognizing receptor known to suppress innate immune cell activation in lung inflammation models. The nanoparticles disclosed herein have been shown to be effective in inhibiting key pathways involved in both the pathogenesis of Siglec-9-mediated diseases.

[0029] Sialic acids are also used as entry points for several viral families, such as influenza A, influenza B, influenza C, SARS-CoV-1, and SARS-CoV-2. The binding receptors on these viruses can be viral capsid segments such as hemagglutinin esterase (viral HE), neuraminidase (viral N), or spike proteins (viral SP) that bind to sialic acid ligands on the surface of host cells to facilitate viral entry into the host cell, or CD147, a sialic acid-binding lectin used for infectious entry by SARS-CoV-2 and Plasmodium falciparum. Binding these sialic acid receptors with decoy ligands can prevent the virus from infecting the host cell and prevent viral particles from leaving the infected cell.

[0030] There is a need for improved compositions and methods for stimulating sialic acid-binding self-associated pattern recognition receptors for the treatment of diseases resulting from acute, chronic, or aberrant immune system activation. There is also a need to prevent these SAMP receptors from being directed by cancer and infectious diseases to evade immune surveillance and attack. The present invention provides nanoparticles that can present ligands that specifically and potently stimulate, block, or antagonize specific Siglec receptors.

[0031] Definitions of specific functional groups and chemical terms are described in more detail below. Chemical elements are listed in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th Ed., inside cover, and specific functional groups are generally defined as described therein. In addition, general principles of organic chemistry and specific functional moieties and reactivities are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th Edition, John Wiley&Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3 rd Edition, Cambridge University Press, Cambridge, 1987.

[0032] The compounds described herein may contain one or more asymmetric centers and therefore may exist in various stereoisomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high-performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions, Wiley Interscience, New York, 1981; Wilen et al., Tetrahedron 33:2725 (1977); Eliel, ELStereochemistry of Carbon Compounds, McGraw-Hill, NY, 1962; and Wilen, SH, Tables of Resolving Agents and Optical Resolutions p. 268, ELS Eliel, Ed., University of Notre Dame Press, Notre Dame, IN 1972. The present invention further encompasses compounds as individual isomers substantially free of other isomers or as mixtures of various isomers.

[0033] In the formula: [ka] is a single bond, and the stereochemistry of the moiety directly bonded thereto is unspecified; --- is absent or a single bond; [ka] is a single bond or a double bond.

[0034] When a range of values ​​is listed, it is intended to encompass each value and subrange within that range. For example, "C 1~6 "Alkyl" refers to C1, C2, C3, C4, C5, C6, C 1~6 , C 1~5 , C 1~4 , C 1~3 , C 1~2 , C 2~6 , C 2~5 , C 2~4 , C 2~3 , C 3~6 , C 3~5 , C 3~4 , C 4~6 , C 4~5 and C 5~6 Alkyl is intended to be included.

[0035] The term "alkyl" refers to the radical of a straight or branched chain saturated hydrocarbon group having the specified range of carbon atoms (e.g., "C 1~16 In some embodiments, an alkyl group has 1 to 9 carbon atoms ("C 1~9 In some embodiments, an alkyl group has 1 to 8 carbon atoms ("C 1~8 In some embodiments, the alkyl group has 1 to 7 carbon atoms ("C alkyl"). In some embodiments, the alkyl group has 1 to 6 carbon atoms ("C 1~6 In some embodiments, an alkyl group has 1 to 5 carbon atoms ("C 1~5In some embodiments, an alkyl group has 1 to 4 carbon atoms ("C 1~4 In some embodiments, an alkyl group has 1 to 3 carbon atoms ("C 1~3 In some embodiments, the alkyl group has 1 to 2 carbon atoms ("C 1~2 In some embodiments, the alkyl group has one carbon atom ("C alkyl"). In some embodiments, the alkyl group has two to six carbon atoms ("C 2~6 alkyl). C 1~6 Examples of alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, iso-butyl), pentyl (C5) (e.g., n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2-butanyl, tertiary amyl), and hexyl (C6) (e.g., n-hexyl). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted ("unsubstituted alkyl") or substituted with one or more substituents (e.g., halogen, such as F) ("substituted alkyl"). In certain embodiments, an alkyl group is an unsubstituted C1 to 10 Alkyl (unsubstituted C 1~6 Alkyl, such as -CH3 (Me), unsubstituted ethyl (Et), unsubstituted propyl (Pr, e.g., unsubstituted n-propyl (n-Pr), unsubstituted isopropyl (i-Pr)), unsubstituted butyl (Bu, e.g., unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl (tert-Bu or t-Bu), unsubstituted sec-butyl (sec-Bu), unsubstituted isobutyl (i-Bu)). In certain embodiments, the alkyl group is a substituted C1 to 10 Alkyl (substituted C 1~6 alkyl, for example, -CF3, Bn, etc.

[0036] The term "alkylenyl" refers to a group having a specific range of carbon atoms (e.g., "C 1~16"Alkyl" refers to a divalent radical of a linear, cyclic, or branched saturated hydrocarbon group (which may have 1 to 16 carbon atoms). An example of an alkylenyl is methylene (-CH-). An alkylenyl can be substituted as described above for alkyl.

[0037] The term "haloalkyl" refers to a substituted alkyl group in which one or more of the hydrogen atoms are independently replaced with a halogen, such as fluoro, bromo, chloro, or iodo. In some embodiments, the haloalkyl moiety has 1 to 8 carbon atoms ("C 1~8 In some embodiments, the haloalkyl moiety has 1 to 6 carbon atoms ("C 1~6 In some embodiments, the haloalkyl moiety has 1 to 4 carbon atoms ("C 1~4 In some embodiments, the haloalkyl moiety has 1 to 3 carbon atoms ("C 1~3 In some embodiments, the haloalkyl moiety has 1 to 2 carbon atoms ("C 1~2 Examples of haloalkyl groups include -CHF2, -CH2F, -CF3, -CH2CF3, -CF2CF3, -CF2CF2CF3, -CCl3, -CFCl2, -CF2Cl, and the like.

[0038] The term "hydroxyalkyl" refers to a substituted alkyl group in which one or more of the hydrogen atoms is independently replaced with a hydroxyl. In some embodiments, the hydroxyalkyl moiety has 1 to 8 carbon atoms ("C 1~8 In some embodiments, the hydroxyalkyl moiety has 1 to 6 carbon atoms ("C 1~6 In some embodiments, the hydroxyalkyl moiety has 1 to 4 carbon atoms ("C 1~4 In some embodiments, the hydroxyalkyl moiety has 1 to 3 carbon atoms ("C 1~3In some embodiments, the hydroxyalkyl moiety has 1 to 2 carbon atoms ("C 1~2 hydroxyalkyl).

[0039] The term "alkoxy" refers to an alkyl group, as defined herein, attached to the parent molecular moiety through an oxygen atom. In some embodiments, the alkoxy moiety has 1 to 8 carbon atoms ("C 1~8 In some embodiments, the alkoxy moiety has 1 to 6 carbon atoms ("C 1~6 In some embodiments, the alkoxy moiety has 1 to 4 carbon atoms ("C 1~4 In some embodiments, the alkoxy moiety has 1 to 3 carbon atoms ("C 1~3 alkoxy).

[0040] In some embodiments, the alkoxy moiety has 1 to 2 carbon atoms ("C 1~2 Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, and tert-butoxy.

[0041] The term "haloalkoxy" refers to a haloalkyl group, as defined herein, attached to the parent molecular moiety through an oxygen atom. In some embodiments, the alkoxy moiety has 1 to 8 carbon atoms ("C 1~8 In some embodiments, the alkoxy moiety has 1 to 6 carbon atoms ("C 1~6 In some embodiments, the alkoxy moiety has 1 to 4 carbon atoms ("C 1~4 In some embodiments, the alkoxy moiety has 1 to 3 carbon atoms ("C 1~3 In some embodiments, the alkoxy moiety has 1 to 2 carbon atoms ("C 1~2Representative examples of haloalkoxy include, but are not limited to, difluoromethoxy, trifluoromethoxy, and 2,2,2-trifluoroethoxy.

[0042] The term "alkoxyalkyl" refers to a substituted alkyl group in which one or more of the hydrogen atoms is independently replaced with an alkoxy group, as defined herein. In some embodiments, the alkoxyalkyl moiety has 1 to 8 carbon atoms ("C 1~8 In some embodiments, the alkoxyalkyl moiety has 1 to 6 carbon atoms ("C 1~6 In some embodiments, the alkoxyalkyl moiety has 1 to 4 carbon atoms ("C 1~4 In some embodiments, the alkoxyalkyl moiety has 1 to 3 carbon atoms ("C 1~3 In some embodiments, the alkoxyalkyl moiety has 1 to 2 carbon atoms ("C 1~2 alkoxyalkyl").

[0043] The term "heteroalkyl" refers to an alkyl group that further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within the parent chain (i.e., inserted between adjacent carbon atoms of the parent chain) and / or disposed at one or more terminal positions of the parent chain. In certain embodiments, a heteroalkyl group is a saturated group having 1 to 20 carbon atoms and one or more heteroatoms within the parent chain ("heteroC 1~20 In some embodiments, a heteroalkyl group refers to a saturated group having 1 to 18 carbon atoms and one or more heteroatoms in the parent chain ("heteroC 1~18 In some embodiments, a heteroalkyl group is a saturated group having 1 to 16 carbon atoms and one or more heteroatoms in the parent chain ("heteroC 1~16 In some embodiments, heteroalkyl groups are saturated groups having 1 to 14 carbon atoms and one or more heteroatoms in the parent chain ("heteroC1~14 In some embodiments, a heteroalkyl group is a saturated group having 1 to 12 carbon atoms and one or more heteroatoms in the parent chain ("heteroC 1~12 In some embodiments, a heteroalkyl group is a saturated group having 1 to 10 carbon atoms and one or more heteroatoms in the parent chain ("heteroC 1~10 In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and one or more heteroatoms in the parent chain ("heteroC 1~8 In some embodiments, a heteroalkyl group is a saturated group having 1 to 6 carbon atoms and one or more heteroatoms in the parent chain ("heteroC 1~6 In some embodiments, heteroalkyl groups are saturated groups having 1 to 4 carbon atoms and 1 or 2 heteroatoms in the parent chain ("heteroC 1~4 In some embodiments, a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom in the parent chain ("heteroC 1~3 In some embodiments, heteroalkyl groups are saturated groups having 1 to 2 carbon atoms and 1 heteroatom in the parent chain ("heteroC 1~2 In some embodiments, a heteroalkyl group is a saturated group having one carbon atom and one heteroatom ("heteroC alkyl"). In some embodiments, a heteroalkyl group, as defined herein, is a partially unsaturated group having one or more heteroatoms and at least one unsaturated carbon in the parent chain, e.g., a carbonyl group. For example, a heteroalkyl group can include an amide or ester functionality in its parent chain such that one or more carbon atoms is an unsaturated carbonyl group. Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted ("unsubstituted heteroalkyl") or substituted with one or more substituents ("substituted heteroalkyl"). In certain embodiments, a heteroalkyl group is an unsubstituted heteroC 1~20 In certain embodiments, the heteroalkyl group is an unsubstituted heteroC1-C1 alkyl. 10In certain embodiments, the heteroalkyl group is a substituted heteroC 1~20 In certain embodiments, the heteroalkyl group is an unsubstituted heteroC1-C1 alkyl. 10 It is alkyl.

[0044] The term "alkenyl" refers to the radical of a straight-chain or branched hydrocarbon group having 2 to 10 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds). In some embodiments, an alkenyl group has 2 to 9 carbon atoms ("C 2~9 In some embodiments, an alkenyl group has 2 to 8 carbon atoms ("C 2~8 In some embodiments, an alkenyl group has 2 to 7 carbon atoms ("C 2~7 In some embodiments, an alkenyl group has 2 to 6 carbon atoms ("C 2~6 In some embodiments, an alkenyl group has 2 to 5 carbon atoms ("C 2~5 In some embodiments, an alkenyl group has 2 to 4 carbon atoms ("C 2~4 In some embodiments, the alkenyl group has 2 to 3 carbon atoms ("C 2~3 In some embodiments, the alkenyl group has two carbon atoms ("C2 alkenyl"). The one or more carbon-carbon double bonds can be internal (e.g., in 2-butenyl) or terminal (e.g., in 1-butenyl). C 2~4 Examples of alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. 2~6 Examples of alkenyl groups include the above-mentioned C 2~4Alkenyl groups include pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (C8), and the like. Unless otherwise specified, each instance of an alkenyl group is independently unsubstituted ("unsubstituted alkenyl") or substituted with one or more substituents ("substituted alkenyl"). In certain embodiments, an alkenyl group is an unsubstituted C 2~10 In certain embodiments, the alkenyl group is a substituted C 2~10 Alkenyl groups are those in which the C=C double bond is unspecified in terms of stereochemistry (e.g., -CH=CHCH3 or [ka] ) can be an (E)- or a (Z)-double bond.

[0045] The term "heteroalkenyl" refers to an alkenyl group that further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within the parent chain (i.e., inserted between adjacent carbon atoms of the parent chain) and / or disposed at one or more terminal positions of the parent chain. In certain embodiments, a heteroalkenyl group is a group having 2 to 10 carbon atoms, at least one double bond, and one or more heteroatoms within the parent chain ("heteroalkenyl"). 2~10 In some embodiments, heteroalkenyl groups have 2 to 9 carbon atoms, at least one double bond, and one or more heteroatoms in the parent chain ("heteroC"). 2~9 alkenyl).

[0046] In some embodiments, heteroalkenyl groups have 2 to 8 carbon atoms, at least one double bond, and one or more heteroatoms in the parent chain ("heteroC"). 2~8 In some embodiments, heteroalkenyl groups have 2 to 7 carbon atoms, at least one double bond, and one or more heteroatoms in the parent chain ("heteroC"). 2~7In some embodiments, heteroalkenyl groups have 2 to 6 carbon atoms, at least one double bond, and one or more heteroatoms in the parent chain ("heteroC"). 2~6 In some embodiments, heteroalkenyl groups have 2 to 5 carbon atoms, at least one double bond, and 1 or 2 heteroatoms in the parent chain ("heteroC 2~5 In some embodiments, heteroalkenyl groups have 2 to 4 carbon atoms, at least one double bond, and 1 or 2 heteroatoms in the parent chain ("heteroC 2~4 In some embodiments, heteroalkenyl groups have 2 to 3 carbon atoms, at least one double bond, and one heteroatom in the parent chain ("heteroC 2~3 In some embodiments, heteroalkenyl groups have 2 to 6 carbon atoms, at least one double bond, and 1 or 2 heteroatoms in the parent chain ("heteroC 2~6 Unless otherwise specified, each instance of a heteroalkenyl group is independently unsubstituted (an "unsubstituted heteroalkenyl") or substituted (a "substituted heteroalkenyl") with one or more substituents. In certain embodiments, a heteroalkenyl group is an unsubstituted heteroC 2~10 In certain embodiments, the heteroalkenyl group is a substituted heteroC 2~10 It is alkenyl.

[0047] The term "alkynyl" refers to the radical of a straight-chain or branched hydrocarbon group having 2 to 10 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) ("C 2~10 In some embodiments, an alkynyl group has 2 to 9 carbon atoms ("C 2~9 In some embodiments, an alkynyl group has 2 to 8 carbon atoms ("C 2~8 In some embodiments, an alkynyl group has 2 to 7 carbon atoms ("C 2~7 In some embodiments, an alkynyl group has 2 to 6 carbon atoms ("C2~6 In some embodiments, an alkynyl group has 2 to 5 carbon atoms ("C 2~5 In some embodiments, an alkynyl group has 2 to 4 carbon atoms ("C 2~4 In some embodiments, an alkynyl group has 2 to 3 carbon atoms ("C 2~3 In some embodiments, the alkynyl group has two carbon atoms ("C2 alkynyl"). The one or more carbon-carbon triple bonds can be internal (e.g., in 2-butynyl) or terminal (e.g., in 1-butynyl). C 2~4 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. 2~6 Examples of alkenyl groups include the above-mentioned C 2~4 Alkynyl groups include pentynyl (C5), hexynyl (C6), and the like. Additional examples of alkynyl include heptynyl (C7), octynyl (C8), and the like. Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted ("unsubstituted alkynyl") or substituted with one or more substituents ("substituted alkynyl"). In certain embodiments, an alkynyl group is an unsubstituted C 2~10 In certain embodiments, the alkynyl group is a substituted C 2~10 It is alkynyl.

[0048] The term "heteroalkynyl" refers to an alkynyl group that further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within the parent chain (i.e., inserted between adjacent carbon atoms of the parent chain) and / or disposed at one or more terminal positions of the parent chain. In certain embodiments, a heteroalkynyl group is a group having 2 to 10 carbon atoms, at least one triple bond, and one or more heteroatoms within the parent chain ("heteroalkynyl"). 2~10 In some embodiments, heteroalkynyl groups have 2 to 9 carbon atoms, at least one triple bond, and one or more heteroatoms in the parent chain ("heteroC").2~9 In some embodiments, heteroalkynyl groups have 2 to 8 carbon atoms, at least one triple bond, and one or more heteroatoms in the parent chain ("heteroC"). 2~8 In some embodiments, heteroalkynyl groups have 2 to 7 carbon atoms, at least one triple bond, and one or more heteroatoms in the parent chain ("heteroC 2~7 In some embodiments, heteroalkynyl groups have 2 to 6 carbon atoms, at least one triple bond, and one or more heteroatoms in the parent chain ("heteroC"). 2~6 In some embodiments, heteroalkynyl groups have 2 to 5 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms in the parent chain ("heteroC 2~5 In some embodiments, heteroalkynyl groups have 2 to 4 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms ("heteroC 2~4 In some embodiments, heteroalkynyl groups have 2 to 3 carbon atoms, at least one triple bond, and one heteroatom in the parent chain ("heteroC 2~3 In some embodiments, heteroalkynyl groups have 2 to 6 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms in the parent chain ("heteroC 2~6 Unless otherwise specified, each instance of a heteroalkynyl group is independently unsubstituted (an "unsubstituted heteroalkynyl") or substituted (a "substituted heteroalkynyl") with one or more substituents. In certain embodiments, a heteroalkynyl group is an unsubstituted heteroC 2~10 In certain embodiments, the heteroalkynyl group is a substituted heteroC 2~10 It is alkynyl.

[0049] The term "carbocyclyl" or "carbocyclic" refers to a ring system having 3 to 14 ring carbon atoms ("C 3~14"Carbocyclyl" refers to the radical of a non-aromatic cyclic hydrocarbon group having 3 to 10 ring carbon atoms ("C 3~10 In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms ("C 3~8 In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms ("C 3~7 In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms ("C 3~6 In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms ("C 4~6 In some embodiments, the carbocyclyl group has 5 to 6 ring carbon atoms ("C 5~6 In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms ("C 5~10 carbocyclyl). Exemplary C 3~6 Carbocyclyl groups include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like.

[0050] Exemplary C 3~8 The carbocyclyl group includes, but is not limited to, the above-mentioned C 3~6 Examples of carbocyclyl groups include cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), and the like. 3~10 The carbocyclyl group includes, but is not limited to, the above-mentioned C 3~8 Carbocyclyl groups and cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C 10 ), cyclodecenyl (C10 ), octahydro-1H-indenyl (C9), decahydronaphthalenyl (C 10 ), spiro[4.5]decanyl (C 10 As the foregoing examples illustrate, in certain embodiments, a carbocyclyl group is either monocyclic ("monocyclic carbocyclyl") or polycyclic (including, for example, fused, bridged, or spiro ring systems such as a bicyclic ring system ("bicyclic carbocyclyl") or a tricyclic ring system ("tricyclic carbocyclyl")) and may be saturated or contain one or more carbon-carbon double or triple bonds. "Carbocyclyl" also includes ring systems in which a carbocyclyl ring, as defined above, is fused to one or more aryl or heteroaryl groups, the point of attachment being on the carbocyclyl ring; in such cases, the carbon numbering continues to designate the number of carbons in the carbocyclyl ring system. Unless otherwise specified, each instance of a carbocyclyl group is independently unsubstituted ("unsubstituted carbocyclyl") or substituted with one or more substituents ("substituted carbocyclyl"). In certain embodiments, a carbocyclyl group is an unsubstituted C 3~14 In certain embodiments, the carbocyclyl group is a substituted C 3~14 It is a carbocyclyl.

[0051] In some embodiments, a "carbocyclyl" has 3 to 14 ring carbon atoms ("C 3~14 In some embodiments, a cycloalkyl group has 3 to 10 ring carbon atoms ("C 3~10 In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms ("C 3~8 In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms ("C 3~6 In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms ("C 4~6 In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms ("C 5~6In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms ("C 5~10 Cycloalkyl). C 5~6 Examples of cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C6). 3~6 Examples of cycloalkyl groups include the aforementioned C 5~6 Cycloalkyl groups include cyclopropyl (C3) and cyclobutyl (C4). 3~8 Examples of cycloalkyl groups include the aforementioned C 3~6 Cycloalkyl groups include cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted ("unsubstituted cycloalkyl") or substituted with one or more substituents ("substituted cycloalkyl"). In certain embodiments, a cycloalkyl group is an unsubstituted C 3~14 In certain embodiments, the cycloalkyl group is a substituted C 3~14 It is cycloalkyl.

[0052] As used herein, the term "heterocyclyl" refers to an aromatic (also called heteroaryl), unsaturated or saturated cyclic hydrocarbon containing at least one heteroatom in the ring.

[0053] For example, the term "heterocyclyl" or "heterocyclic" refers to the radical of a 3- to 14-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, each independently selected from nitrogen, oxygen, and sulfur (a "3- to 14-membered heterocyclyl"). In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be at a carbon or nitrogen atom, where valence permits. Heterocyclyl groups may be either monocyclic (a "monocyclic heterocyclyl") or polycyclic (e.g., fused, bridged, or spiro ring systems such as bicyclic ring systems (a "bicyclic heterocyclyl") or tricyclic ring systems (a "tricyclic heterocyclyl")), and may be saturated or contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems may contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which a heterocyclyl ring as defined above is fused to one or more carbocyclyl groups (the point of attachment is on either the carbocyclyl or heterocyclyl ring) or to one or more aryl or heteroaryl groups (the point of attachment is on the heterocyclyl ring); in such cases, the numbering of ring members continues to designate the numbering of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl is independently unsubstituted (an "unsubstituted heterocyclyl") or substituted with one or more substituents (a "substituted heterocyclyl"). In certain embodiments, a heterocyclyl group is an unsubstituted 3- to 14-membered heterocyclyl. In certain embodiments, a heterocyclyl group is a substituted 3- to 14-membered heterocyclyl.

[0054] In some embodiments, a heterocyclyl group is a 5- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, each independently selected from nitrogen, oxygen, and sulfur (a "5- to 10-membered heterocyclyl"). In some embodiments, a heterocyclyl group is a 5- to 8-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, each independently selected from nitrogen, oxygen, and sulfur (a "5- to 8-membered heterocyclyl"). In some embodiments, a heterocyclyl group is a 5- to 6-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, each independently selected from nitrogen, oxygen, and sulfur (a "5- to 6-membered heterocyclyl"). In some embodiments, a 5- to 6-membered heterocyclyl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5- to 6-membered heterocyclyl has 1 to 2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heterocyclyl has one ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0055] Exemplary 3-membered heterocyclyl groups containing one heteroatom include, but are not limited to, aziridinyl, oxiranyl, and thiiranyl. Exemplary 4-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclyl groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groups containing two heteroatoms include, but are not limited to, dioxolanyl, oxathiolanyl, and dithiolanyl. Exemplary 5-membered heterocyclyl groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclyl groups containing three heteroatoms include, but are not limited to, triazinyl. Exemplary 7-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azepanyl, oxepanyl, and thiepanyl. Exemplary 8-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azocanyl, oxecanyl, and thiocanyl.Exemplary bicyclic heterocyclyl groups include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl, phthalimidyl, naphthalimidyl, chromanyl, chromenyl, 1H-benzo[e][1,4]diazepine ... nyl, 1,4,5,7-tetrahydropyrano[3,4-b]pyrrolyl, 5,6-dihydro-4H-furo[3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H-pyrrolo[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofuro[3,2-c]pyridinyl, 4,5,6,7-tetrahydrothieno[3,2-b]pyridinyl, 1,2,3,4-tetrahydro-1,6-naphthyridinyl, and the like.

[0056] The term "aryl" refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 pi electrons shared within the cyclic array) having 6 to 14 ring carbon atoms and 0 heteroatoms provided in the aromatic ring system ("C 6~14 In some embodiments, an aryl group has 6 ring carbon atoms ("C aryl"; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C 10 Aryl"; e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("C 14"Aryl"; e.g., anthracyl). "Aryl" also includes ring systems in which an aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups, where the radical or point of attachment is on the aryl ring, and in such cases the carbon numbering continues to designate the number of carbons in the aryl ring system. Unless otherwise specified, each instance of an aryl group is independently unsubstituted ("unsubstituted aryl") or substituted with one or more substituents ("substituted aryl"). In certain embodiments, an aryl group is an unsubstituted C 6~14 In certain embodiments, the aryl group is a substituted C 6~14 It is aryl.

[0057] "Aralkyl" is a subset of "alkyl" and refers to an alkyl group substituted with an aryl group, where the point of attachment is on the alkyl portion.

[0058] The term "heteroaryl" refers to the radical of a 5- to 14-membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared within the cyclic array) having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, each heteroatom being independently selected from nitrogen, oxygen, and sulfur ("5- to 14-membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be at a carbon or nitrogen atom, valence permitting. Heteroaryl polycyclic ring systems may contain one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which a heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups, the point of attachment being on the heteroaryl ring; in such cases, the numbering of the ring members continues to designate the numbering of the ring members within the heteroaryl ring system. "Heteroaryl" also includes ring systems in which a heteroaryl ring, as defined above, is fused to one or more aryl groups, and the point of attachment can be on either the aryl or heteroaryl ring; in such cases, the ring member numbering continues to designate the ring member numbering within the fused polycyclic (aryl / heteroaryl) ring system. In polycyclic heteroaryl groups in which one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.), the point of attachment can be on either ring, i.e., on the ring containing a heteroatom (e.g., 2-indolyl) or on the ring without a heteroatom (e.g., 5-indolyl).

[0059] In some embodiments, a heteroaryl group is a 5- to 10-membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, each heteroatom independently selected from nitrogen, oxygen, and sulfur ("5- to 10-membered heteroaryl"). In some embodiments, a heteroaryl group is a 5- to 8-membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, each heteroatom independently selected from nitrogen, oxygen, and sulfur ("5- to 8-membered heteroaryl"). In some embodiments, a heteroaryl group is a 5- to 6-membered aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms provided in the aromatic ring system, each heteroatom independently selected from nitrogen, oxygen, and sulfur ("5- to 6-membered heteroaryl"). In some embodiments, a 5- to 6-membered heteroaryl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5- to 6-membered heteroaryl has 1 to 2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heteroaryl has one ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted ("unsubstituted heteroaryl") or substituted with one or more substituents ("substituted heteroaryl"). In certain embodiments, the heteroaryl group is an unsubstituted 5- to 14-membered heteroaryl. In certain embodiments, the heteroaryl group is a substituted 5- to 14-membered heteroaryl.

[0060] Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pterinidyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplary tricyclic heteroaryl groups include, but are not limited to, phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl, and phenazinyl.

[0061] "Heteroaralkyl" is a subset of "alkyl" and refers to an alkyl group substituted with a heteroaryl group, where the point of attachment is on the alkyl portion.

[0062] The suffix "-ene" added to a group indicates that the group is a divalent moiety, for example, alkylene is a divalent moiety of alkyl, alkenylene is a divalent moiety of alkenyl, alkynylene is a divalent moiety of alkynyl, heteroalkylene is a divalent moiety of heteroalkyl, heteroalkenylene is a divalent moiety of heteroalkenyl, heteroalkynylene is a divalent moiety of heteroalkynyl, carbocyclylene is a divalent moiety of carbocyclyl, heterocyclylene is a divalent moiety of heterocyclyl, arylene is a divalent moiety of aryl, and heteroarylene is a divalent moiety of heteroaryl.

[0063] A group is optionally substituted unless expressly specified otherwise. The term "optionally substituted" refers to substituted or unsubstituted. In certain embodiments, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups are optionally substituted. "Optionally substituted" refers to a group that can be substituted or unsubstituted (e.g., "substituted" or "unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl, "substituted" or "unsubstituted" alkynyl, "substituted" or "unsubstituted" heteroalkyl, "substituted" or "unsubstituted" heteroalkenyl, "substituted" or "unsubstituted" heteroalkynyl, "substituted" or "unsubstituted" carbocyclyl, "substituted" or "unsubstituted" heterocyclyl, "substituted" or "unsubstituted" aryl, or "substituted" or "unsubstituted" heteroaryl group). In general, the term "substituted" means that at least one hydrogen atom present on a group is replaced with an acceptable substituent, e.g., a substituent that, upon substitution, results in a stable compound, e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise specified, a "substituted" group has a substituent at one or more substitutable positions of the group; when more than one position in any given structure is substituted, the substituents may be the same or different at each position. The term "substituted" includes substitution with all permissible substituents of organic compounds and is intended to include any of the substituents described herein that result in the formation of a stable compound. The present invention contemplates any and all such combinations to arrive at a stable compound. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any suitable substituents, as described herein, that satisfy the valence of the heteroatom and result in the formation of a stable moiety. The present invention is in no way limited by the exemplary substituents described herein.

[0064] Exemplary carbon atom substituents are halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR aa , -ON(R bb)2、-N(R bb )2、-N(R bb )3 + X - 、-N(OR cc )R bb 、-SH、-SR aa 、-SSR cc 、-C(=O)R aa 、-CO2H、-CHO、-C(OR cc )3、-CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(R bb )2、-OC(=O)N(R bb )2、-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2、-C(=O)NR bb SO2R aa 、-NR bb SO2R aa 、-SO2N(R bb )2、-SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3、-OSi(R aa )3、-C(=S)N(R bb )2、-C(=O)SR aa 、-C(=S)SRaa , -SC(=S)SR aa , -SC(=O)SR aa , -OC(=O)SR aa , -SC(=O)OR aa , -SC(=O)R aa , -P(=O)(R aa )2, -P(=O)(OR cc )2, -OP(=O)(R aa )2, -OP(=O)(OR cc )2, -P(=O)(N(R bb )2)2, -OP(=O)(N(R bb )2)2, -NR bb P(=O)(R aa )2, -NR bb P(=O)(OR cc )2, -NR bb P(=O)(N(R bb )2)2, -P(R cc )2, -P(OR cc )2, -P(R cc )3 + X - , -P(OR cc )3 + X - , -P(R cc )4, -P(OR cc )2, -OP(R cc )2, -OP(R cc )3 + X - , -OP(OR cc )2, -OP(OR cc )3 + X - , -OP(R cc )4, -OP(OR cc )4, -B(R aa )2, -B(OR cc )2, -BR aa (OR cc ), C 1~10 Alkyl, C 1~10 Perhaloalkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, Hetero C 1~10 Alkyl, Hetero C 2~10 Alkenyl, Hetero C 2~10 Alkynyl, C3~10 Carbocyclyl, 3- to 14-membered heterocyclyl, C 6~14 Each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R dd independently substituted with groups; X - is a counterion; or the two geminal hydrogens on the carbon atom are ═O, ═S, ═NN(R bb )2, =NNR bb C(=O)R aa , =NNR bb C(=O)OR aa , =NNR bb S(=O)2R aa , =NR bb or =NOR cc substituted with R aa Each example of C 1~10 Alkyl, C 1~10 Haloalkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, Hetero C 1~10 Alkyl, Hetero C 2~10 Alkenyl, Hetero C 2~10 Alkynyl, C 3~10 Carbocyclyl, 3- to 14-membered heterocyclyl, C 6~14 aryl and 5- to 14-membered heteroaryl, or two R aa The groups may be joined to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R dd independently substituted with R groups; bb Examples of each are hydrogen, -OH, -OR aa , -N(R cc )2, -CN, -C(=O)R aa , -C(=O)N(R cc )2, -CO2R aa , -SO2R aa , -C(=NRcc ) OR aa , -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc , -SOR aa , -C(=S)N(R cc )2, -C(=O)SR cc , -C(=S)SR cc , -P(=O)(R aa )2, -P(=O)(OR cc )2, -P(=O)(N(R cc )2)2, C 1~10 Alkyl, C 1~10 Perhaloalkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, Hetero C 1~10 Alkyl, Hetero C 2~10 Alkenyl, Hetero C 2~10 Alkynyl, C 3~10 Carbocyclyl, 3- to 14-membered heterocyclyl, C 6~14 aryl and 5- to 14-membered heteroaryl, or two R bb The groups may be joined to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R dd independently substituted with groups; X - is the counterion; R cc Each example of is independently hydrogen, C1 10 Alkyl, C1~ 10 Perhaloalkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, hetero C1 10 Alkyl, Hetero C 2~10 Alkenyl, Hetero C 2~10 Alkynyl, C 3~10 Carbocyclyl, 3- to 14-membered heterocyclyl, C 6~14 aryl and 5- to 14-membered heteroaryl, or two R ccThe groups may be joined to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R dd independently substituted with R groups; dd Each example is independently a halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee , -ON(R ff )2, -N(R ff )2, -N(R ff )3 + X - , -N(OR ee )R ff , -SH, -SR ee , -SSR ee , -C(=O)R ee , -CO2H, -CO2R ee , -OC(=O)R ee , -OCO2R ee , -C(=O)N(R ff )2, -OC(=O)N(R ff )2, -NR ff C(=O)R ee , -NR ff CO2R ee , -NR ff C(=O)N(R ff )2, -C(=NR ff ) OR ee , -OC(=NR ff )R ee , -OC(=NR ff ) OR ee , -C(=NR ff )N(R ff )2, -OC(=NR ff )N(R ff )2, -NR ff C(=NR ff )N(R ff )2, -NR ff SO2R ee , -SO2N(R ff )2, -SO2R ee , -SO2OR ee , -OSO2R ee, -S(=O)R ee , -Si(R ee )3, -OSi(R ee )3, -C(=S)N(R ff )2, -C(=O)SR ee , -C(=S)SR ee , -SC(=S)SR ee , -P(=O)(OR ee )2, -P(=O)(R ee )2, -OP(=O)(R ee )2, -OP(=O)(OR ee )2, C 1~6 Alkyl, C 1~6 Perhaloalkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, Hetero C 1~6 Alkyl, Hetero C 2~6 Alkenyl, Hetero C 2~6 Alkynyl, C 3~10 Carbocyclyl, 3-10 membered heterocyclyl, C 6~10 aryl, and 5- to 10-membered heteroaryl, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R gg groups, or two geminal R dd The substituents can be linked to form =O or =S; X - is the counterion; R ee Each example of C 1~6 Alkyl, C 1~6 Perhaloalkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, Hetero C 1~6 Alkyl, Hetero C 2~6 Alkenyl, Hetero C 2~6 Alkynyl, C 3~10 Carbocyclyl, C 6~10aryl, 3- to 10-membered heterocyclyl, and 3- to 10-membered heteroaryl, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may be selected from 0, 1, 2, 3, 4, or 5 R gg independently substituted with R groups; ff Each instance of is independently hydrogen, C 1~6 Alkyl, C 1~6 Perhaloalkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, Hetero C 1~6 Alkyl, Hetero C 2~6 Alkenyl, Hetero C 2~6 Alkynyl, C 3~10 Carbocyclyl, 3-10 membered heterocyclyl, C 6~10 aryl and 5- to 10-membered heteroaryl, or two R ff groups are joined to form a 3- to 10-membered heterocyclyl or 5- to 10-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R gg independently substituted with R groups; gg Each example is independently a halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC 1~6 Alkyl, -ON(C 1~6 alkyl)2, -N(C 1~6 alkyl)2, -N(C 1~6 alkyl)3 + X - , -NH(C 1~6 alkyl)2 + X - , -NH2(C 1~6 alkyl) + X - , -NH3 + X - , -N(OC 1~6 Alkyl)(C 1~6 alkyl), -N(OH)(C 1~6 alkyl), -NH(OH), -SH, -SC 1~6 Alkyl, -SS(C1~6 alkyl), -C(=O)(C 1~6 alkyl), -CO2H, -CO2(C 1~6 alkyl), -OC(=O)(C 1~6 alkyl), -OCO2(C 1~6 alkyl), -C(=O)NH2, -C(=O)N(C 1~6 alkyl)2, -OC(=O)NH(C 1~6 alkyl), -NHC(=O)(C 1~6 alkyl), -N(C 1~6 alkyl)C(=O)(C 1~6 alkyl), -NHCO2(C 1~6 alkyl), -NHC(=O)N(C 1~6 alkyl)2, -NHC(=O)NH(C 1~6 alkyl), -NHC(=O)NH2, -C(=NH)O(C 1~6 alkyl), -OC(=NH)(C 1~6 alkyl), -OC(=NH)OC 1~6 Alkyl, -C(=NH)N(C 1~6 alkyl)2, -C(=NH)NH(C 1~6 alkyl), -C(=NH)NH2, -OC(=NH)N(C 1~6 alkyl)2, -OC(=NH)NH(C 1~6 alkyl), -OC(=NH)NH2, -NHC(=NH)N(C 1~6 alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1~6 alkyl), -SO2N(C 1~6 alkyl)2, -SO2NH(C 1~6 alkyl), -SO2NH2, -SO2(C 1~6 alkyl), -SO2O(C 1~6 alkyl), -OSO2(C 1~6 alkyl), -SO(C 1~6 alkyl), -Si(C 1~6 alkyl)3, -OSi(C 1~6 alkyl)3, -C(=S)N(C 1~6 alkyl)2, -C(=S)NH(C 1~6 alkyl), -C(=S)NH2, -C(=O)S(C 1~6 alkyl), -C(=S)SC1~6 Alkyl, -SC(=S)SC 1~6 Alkyl, -P(=O)(OC 1~6 alkyl)2, -P(=O)(C 1~6 alkyl)2, -OP(=O)(C 1~6 alkyl)2, -OP(=O)(OC 1~6 Alkyl)2, C 1~6 Alkyl, C 1~6 Perhaloalkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, Hetero C 1~6 Alkyl, Hetero C 2~6 Alkenyl, Hetero C 2~6 Alkynyl, C 3~10 Carbocyclyl, C 6~10 aryl, 3- to 10-membered heterocyclyl, 5- to 10-membered heteroaryl; or two geminal R gg The substituents can be linked to form =O or =S; X - is the counter ion.

[0065] The term "halo" or "halogen" refers to fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br) or iodine (iodo, -I).

[0066] The term "hydroxyl" or "hydroxy" refers to an -OH group. By extension, the term "substituted hydroxyl" or "substituted hydroxyl" refers to a hydroxyl group in which the oxygen atom directly attached to the parent molecule has been replaced with a group other than hydrogen, OR aa , -ON(R bb )2, -OC(=O)SR aa , -OC(=O)R aa , -OCO2R aa , -OC(=O)N(R bb )2, -OC(=NR bb )R aa , -OC(=NR bb ) OR aa , -OC(=NR bb )N(R bb )2, -OS(=O)R aa , -OSO2R aa , -OSi(R aa)3, -OP(R cc )2, -OP(R cc )3 + X - , -OP(OR cc )2, -OP(OR cc )3 + X - , -OP(=O)(R aa )2, -OP(=O)(OR cc )2 and -OP(=O)(N(R bb )2)2(wherein, X - , R aa , R bb and R cc is as described herein).

[0067] The term "amino" refers to the group -NH. By extension, the term "substituted amino" refers to a mono-, di-, or tri-substituted amino. In certain embodiments, a "substituted amino" is a mono- or di-substituted amino group.

[0068] The term "monosubstituted amino" refers to an amino group in which the nitrogen atom directly attached to the parent molecule is replaced with one hydrogen and one non-hydrogen group, and includes -NH(R bb ), -NHC(=O)R aa , -NHCO2R aa , -NHC(=O)N(R bb )2, -NHC(=NR bb )N(R bb )2, -NHSO2R aa , -NHP(=O)(OR cc )2 and -NHP(=O)(N(R bb )2)2(wherein, R aa , R bb and R cc is as described herein, and —NH(R bb )R bb is not hydrogen).

[0069] The term "disubstituted amino" refers to an amino group in which the nitrogen atom directly attached to the parent molecule is replaced with two groups other than hydrogen, -N(R bb )2, -NRbb C(=O)R aa , -NR bb CO2R aa , -NR bb C(=O)N(R bb )2, -NR bb C(=NR bb )N(R bb )2, -NR bb SO2R aa , -NR bb P(=O)(OR cc )2 and -NR bb P(=O)(N(R bb )2)2(wherein, R aa , R bb and R cc is as described herein, except that the nitrogen atom directly attached to the parent molecule is not replaced with a hydrogen.

[0070] The term "trisubstituted amino" refers to an amino group in which the nitrogen atom directly attached to the parent molecule is substituted with three groups, -N(R bb )2 and -N(R bb )3 + X - , (where R bb and X - is as defined herein.

[0071] The term "sulfonyl" refers to -SO2N(R bb )2, -SO2R aa and SO2OR aa (In the formula, R aa and R bb refers to a group selected from:

[0072] The term "sulfinyl" refers to -S(=O)R aa group (in the formula, R aa refers to a compound as defined herein.

[0073] The term "acyl" refers to a group having the general formula -C(=O)R X1 , -C(=O)OR X1, -C(=O)-OC(=O)R X1 , -C(=O)SR X1 , -C(=O)N(R X1 )2, -C(=S)R X1 , -C(=S)N(R X1 )2, -C(=S)O(R X1 ), -C(=S)S(R X1 ), -C(=NR X1 )R X1 , -C(=NR X1 ) OR X1 , -C(=NR X1 )SR X1 and -C(=NR X1 )N(R X1 )2, and R X1 is hydrogen; halogen; substituted or unsubstituted hydroxyl; substituted or unsubstituted thiol; substituted or unsubstituted amino; substituted or unsubstituted acyl, cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched alkyl; cyclic or acyclic, substituted or unsubstituted, branched or unbranched alkenyl; substituted or unsubstituted alkynyl; substituted or unsubstituted aryl, substituted or unsubstituted is unsubstituted heteroaryl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, mono- or di-aliphaticamino, mono- or di-heteroaliphaticamino, mono- or di-alkylamino, mono- or di-heteroalkylamino, mono- or di-arylamino, or mono- or di-heteroarylamino; or two R X1 The groups taken together form a 5- to 6-membered heterocyclic ring.

[0074] Exemplary acyl groups include aldehydes (-CHO), carboxylic acids (-COH), ketones, acyl halides, esters, amides, imines, carbonates, carbamates, and ureas. Acyl substituents include, but are not limited to, any of the substituents described herein that result in the formation of a stable moiety (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thioxo, cyano, isocyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphatic amino, heteroaliphatic amino, alkylamino, heteroalkylamino, arylamino, heteroarylamino, alkylaryl, arylalkyl, aliphaticoxy, heteroaliphaticoxy, alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, aliphaticthioxy, heteroaliphaticthioxy, alkylthioxy, heteroalkylthioxy, arylthioxy, heteroarylthioxy, acyloxy, etc., each of which may or may not be further substituted).

[0075] The term "carbonyl" refers to the carbon directly attached to the parent molecule being sp 2 Groups that are hybridized and substituted with oxygen, nitrogen or sulfur atoms, such as ketones (e.g., -C(=O)R aa ), carboxylic acids (e.g., -COH), aldehydes (CHO), esters (e.g., -COR aa , -C(=O)SR aa , -C(=S)SR aa ), amides (e.g., -C(=O)N(R bb )2, C(=O)NR bb SO2R aa , -C(=S)N(R bb )2 and imines (e.g., -C(=NR bb )R aa , -C(=NR bb ) OR aa ), C(=NR bb )N(R bb )2(wherein, R aa and R bb refers to a group selected from:

[0076] The term "oxo" refers to the group =O and the term "thiooxo" refers to the group =S.

[0077] The term "cyano" refers to the group --CN.

[0078] The term "azido" refers to the group -N3.

[0079] Nitrogen atoms can be substituted or unsubstituted, where valence allows, and include primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents are hydrogen, -OH, -OR aa , -N(R cc )2, -CN, -C(=O)R aa , -C(=O)N(R cc )2, -CO2R aa , -SO2R aa , -C(=NR bb )R aa , -C(=NR cc ) OR aa , -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc , -SOR aa , -C(=S)N(R cc )2, -C(=O)SR cc , -C(=S)SR cc , -P(=O)(OR cc )2, -P(=O)(R aa )2, -P(=O)(N(R cc )2)2, C 1~10 Alkyl, C 1~10 Perhaloalkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, hetero C1 10 Alkyl, Hetero C 2~10 Alkenyl, Hetero C 2~10 Alkynyl, C 3~10 Carbocyclyl, 3- to 14-membered heterocyclyl, C 6~14including, but not limited to, aryl and 5-14 membered heteroaryl, or two R cc The groups are joined to form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R dd groups, and R aa , R bb , R cc and R dd is as described herein.

[0080] In certain embodiments, the substituent present on a nitrogen atom is a nitrogen protecting group (also referred to herein as an "amino protecting group"). Nitrogen protecting groups include -OH, -OR aa , -N(R cc h, -C(=O)R aa , -C(=O)N(R cc )2, -CO2R aa , -SO2R aa , -C(=NR cc )R aa , -C(=NR cc ) OR aa , -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc , -SOR aa , -C(=S)N(R cc )2, -C(=O)SR cc , -C(=S)SR cc , C 1~10 Alkyl (e.g., aralkyl, heteroaralkyl), C 2~10 Alkenyl, C 2~10 Alkynyl, Hetero C 1~10 Alkyl, Hetero C 2~10 Alkenyl, Hetero C 2~10 Alkynyl, C 3~10 Carbocyclyl, 3- to 14-membered heterocyclyl, C 6~14Each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aralkyl, aryl, and heteroaryl may have 0, 1, 2, 3, 4, or 5 R dd groups, and R aa , R bb , R cc and R dd are as described herein. Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Butts, 3rd edition, John Wiley & Sons, 1999 (incorporated herein by reference).

[0081] For example, a nitrogen protecting group such as an amide group (e.g., —C(═O)R aa ) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxyacylaminoacetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinamide, N-acetylmethionine derivatives, o-nitrobenzamide, and o(benzoyloxymethyl)benzamide.

[0082] Nitrogen protecting groups such as carbamate groups (e.g., -C(=O)OR aa) are methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfa)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluorenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl Carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc) ), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC or Boc), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl Dithiocarbamates, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenylcarbamate (Mtpc), 2,4-dimethylthiophenylcarbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chloro Monylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, t-amyl carbamate, S-benzylthiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2, 2-dimethoxyacyl vinyl carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isobornyl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-Methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate and 2,4,Examples include, but are not limited to, 6-trimethylbenzyl carbamate.

[0083] Sulfonamide groups (e.g., -S(=O)R aa Nitrogen protecting groups such as p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), and 2,4,6-trimethylbenzenesulfonamide (Mts) , 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), -trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0084] Other nitrogen protecting groups include phenothiazinyl-(10) acyl derivatives, N'-p-toluenesulfonylaminoacyl derivatives, N'-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, and N-1,1,4,4-tetramethyldisilylazacyclo Pentane adducts (STA bases), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-ones, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-2-ones, 1-substituted 3,5-dinitro-4-pyridones, N-methylamines, N-allylamines, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl)amine, Quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fern), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethylamine N-Benzylideneamine, Np-Methoxybenzylideneamine, N-Diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N',N'-Dimethylaminomethylene)amine, N,N'-Isopropylidenediamine, Np-Nitrobenzylideneamine, N-Salicylideneamine, N-5-Chlorsalicylideneamine, N-(5-Chloro-2-hydroxyphenyl)phenylmethyleneamine, N-Cyclohexylideneamine, N-(5,These include, but are not limited to, 5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylborinic acid derivatives, N[phenyl(pentaacychromium- or tungsten)acyl]amine, N-copper chelates, N-zinc chelates, N-nitroamines, N-nitrosamines, amine N-oxides, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridine sulfenamide (Npys). In certain embodiments, the nitrogen protecting group is benzyl (Bn), tert-butyloxycarbonyl (BOC), carbobenzyloxy (Cbz), 9-flurenylmethyloxycarbonyl (Fmoc), trifluoroacetyl, triphenylmethyl, acetyl (Ac), benzoyl (Bz), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), 2,2,2-trichloroethyloxycarbonyl (Troc), triphenylmethyl (Tr), tosyl (Ts), brosyl (Bs), nosyl (Ns), mesyl (Ms), triflyl (Tf), or dansyl (Ds).

[0085] In certain embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also referred to herein as a "hydroxyl protecting group"). An oxygen protecting group is -R aa , -N(R bb )2, -C(=O)SR aa , -C(=O)R aa , -CO2R aa , -C(=O)N(R bb )2, -C(=NR bb )R aa , -C(=NR bb ) OR aa , -C(=NR bb )N(Rbb )2, -S(=O)R aa , -SO2R aa , -Si(R aa )3, -P(R cc )2, -P(R cc )3 + X - , -P(OR cc )2, -P(OR cc )3 + X - , -P(=O)(R aa )2, -P(=O)(OR cc )2 and -P(=O)(N(R bb )2)2, including but not limited to, X - , R aa , R bb and R cc are as described herein. Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Butts, 3rd edition, John Wiley & Sons, 1999 (incorporated herein by reference).

[0086] Exemplary oxygen protecting groups are methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-Trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4- Methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl , 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, a-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tris(levulinoyloxyphenyl)methyl, 4,4',4"-tris(benzoyloxyphenyl)methyl ) methyl, 3-(imidazol-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylsilyl isopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TEMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetic acid, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate , phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyl dithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), ethyl carbonate, 2,2,2-Trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), isobutyl carbonate, vinyl carbonate, allyl carbonate, t-butyl carbonate (BOC or Boc), p-nitrophenyl carbonate, benzyl carbonate, p- Methoxybenzyl carbonate, 3,4-dimethoxybenzyl carbonate, o-nitrobenzyl carbonate, p-nitrobenzyl carbonate, S-benzylthiocarbonate, 4-ethoxy-1-naphthotyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2- These include, but are not limited to, (methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o(methoxyacyl)benzoate, a-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). In certain embodiments, the oxygen protecting group is silyl. In certain embodiments, the oxygen protecting group is t-butyldiphenylsilyl (TBDPS), t-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS), triphenylsilyl (TPS), triethylsilyl (TES), trimethylsilyl (TMS), triisopropylsiloxymethyl (TOM), acetyl (Ac), benzoyl (Bz), allyl carbonate, 2,2,2-trichloroethyl carbonate (Troc), 2-trimethylsilylethyl carbonate, methoxymethyl (MOM), 1-ethoxyethyl (EE), 2-methoxy-2-propyl (MOP), 2,2,2-trichloroethoxyethyl, 2-methoxyethoxymethyl (MEM), 2-trimethylsilylethoxymethyl (SEM), methylthiomethyl (MTM), tetrahydropyranyl (THP), tetrahydrofuranyl (THF), p-methoxyphenyl (PMP), triphenylmethyl (Tr), methoxytrityl (MMT), dimethoxytrityl (DMT), allyl, p-methoxybenzyl (PMB), t-butyl, benzyl (Bn), allyl, or pivaloyl (Piv).

[0087] In certain embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also referred to herein as a "hydroxyl protecting group"). An oxygen protecting group is -R aa , -N(R bb )2, -C(=O)SR aa , -C(=O)R aa , -CO2R aa , -C(=O)N(R bb )2, -C(=NR bb )R aa , -C(=NR bb ) OR aa , -C(=NR bb )N(R bb )2, -S(=O)R aa , -SO2R aa , -Si(R aa )3, -P(R cc )2, -P(R cc )3 + X - , -P(OR cc )2, -P(OR cc )3 + X - , -P(=O)(R aa )2, -P(=O)(OR cc )2 and -P(=O)(N(R bb )2)2, including but not limited to, X - , R aa , R bb and R ccare as described herein. Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Butts, 3rd edition, John Wiley & Sons, 1999 (incorporated herein by reference).

[0088] Exemplary oxygen protecting groups are methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-Trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4- Methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl , 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, a-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tris(levulinoyloxyphenyl)methyl, 4,4',4"-tris(benzoyloxyphenyl)methyl ) methyl, 3-(imidazol-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylsilyl isopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TEMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetic acid, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate , phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyl dithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), ethyl carbonate, 2,2,2-Trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), isobutyl carbonate, vinyl carbonate, allyl carbonate, t-butyl carbonate (BOC or Boc), p-nitrophenyl carbonate, benzyl carbonate, p- Methoxybenzyl carbonate, 3,4-dimethoxybenzyl carbonate, o-nitrobenzyl carbonate, p-nitrobenzyl carbonate, S-benzylthiocarbonate, 4-ethoxy-1-naphthotyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2- These include, but are not limited to, (methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o(methoxyacyl)benzoate, a-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). In certain embodiments, the oxygen protecting group is silyl. In certain embodiments, the oxygen protecting group is t-butyldiphenylsilyl (TBDPS), t-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS), triphenylsilyl (TPS), triethylsilyl (TES), trimethylsilyl (TMS), triisopropylsiloxymethyl (TOM), acetyl (Ac), benzoyl (Bz), allyl carbonate, 2,2,2-trichloroethyl carbonate (Troc), 2-trimethylsilylethyl carbonate, methoxymethyl (MOM), 1-ethoxyethyl (EE), 2-methoxy-2-propyl (MOP), 2,2,2-trichloroethoxyethyl, 2-methoxyethoxymethyl (MEM), 2-trimethylsilylethoxymethyl (SEM), methylthiomethyl (MTM), tetrahydropyranyl (THP), tetrahydrofuranyl (THF), p-methoxyphenyl (PMP), triphenylmethyl (Tr), methoxytrityl (MMT), dimethoxytrityl (DMT), allyl, p-methoxybenzyl (PMB), t-butyl, benzyl (Bn), allyl, or pivaloyl (Piv).

[0089] In certain embodiments, the substituent present on the sulfur atom is a sulfur protecting group (also referred to as a "thiol protecting group"). A sulfur protecting group is a -R aa , -N(R bb )2, -C(=O)SR aa , -C(=O)R aa , -CO2R aa , -C(=O)N(R bb )2, -C(=NR bb )R aa , -C(=NR bb ) OR aa , -C(=NR bb )N(R bb )2, -S(=O)R aa , -SO2R aa , -Si(R aa )3, -P(R cc )2, -P(R cc )3 + X - , -P(OR cc )2, -P(OR cc )3 + X - , -P(=O)(R aa )2, -P(=O)(OR cc )2 and -P(=O)(N(R bb )2)2, including but not limited to, R aa , R bb and R ccis as described herein. Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Buts, 3rd edition, John Wiley & Sons, 1999 (incorporated herein by reference). In certain embodiments, the sulfur protecting group is acetamidomethyl, t-Bu, 3-nitro-2-pyridinesulfenyl, 2-pyridinesulfenyl, or triphenylmethyl.

[0090] Additionally, the following terms are used herein: · "Ac" refers to an acyl group; "Fmoc" refers to the fluorenylmethoxycarbonyl protecting group; "tBoc" refers to the tert-butyloxycarbonyl protecting group; "Cbz" refers to the benzyl chlorocarbonate protecting group; "Troc" refers to the trichloroethyl chloroformate protecting group; "TFA" refers to a trifluoroacetamide protecting group; The "[1,3]dioxolo group" has the following structural formula: [ka] It refers to the part having

[0091] As used herein, the term "click chemistry reagent" (used interchangeably with "click reagent" or "click moiety") refers to any of two chemical moieties, referred to as a "click reagent pair," that react with its corresponding click moiety via an electrocycloaddition reaction ("click chemistry reaction" or "click reaction") to produce a "click reaction product." Typically, click reactions are performed under mild conditions in aqueous solvents and can include any one of the following: neutral pH, ambient temperature, and low reactant concentrations. Exemplary click reagent pairs include, but are not limited to, Cu(I)-catalyzed azide-alkyne click chemistry reaction (CuAAC); strain-promoted azide-alkyne click chemistry reaction (SPAAC), and tetrazine and alkene (trans-cyclooctene).

[0092] Any suitable click reagent pair can be used in the present invention. Examples of click reagents include: bicyclo[6.1.0]nonyne (BCN) groups, for example [ka] Reagents including; - Alkynes, e.g. [ka] Reagents including; - azides, e.g. N3-CH2-containing reagents; dibenzocyclooctyne (DBCO) groups, for example [ka] Reagents including; trans-cyclooctene, for example [ka] Reagents including; - tetrazines, e.g. [ka] A reagent comprising:

[0093] "Sialic acid" refers to neuraminic acid or any chemical modification of neuraminic acid, whether naturally occurring or synthetically derived. The structural formula of neuraminic acid is reproduced below: [ka]

[0094] As used herein, the terms "poly(ethylene glycol)", "polyethylene oxide" and "polyoxyethylene" are used interchangeably and are defined by the following structural formula: H-(O-CHCH) m It refers to a polymer having an -OH group, where m is the number of repeating units. The polymer molecular weight can vary from 300 g / mol to 10,000,000 g / mol.

[0095] As used herein, the term "particle" includes microparticles and nanoparticles as defined herein.

[0096] As used herein, the term "particle" includes microparticles and nanoparticles as defined herein.

[0097] As used herein, "subject" refers to a subject receiving treatment according to the provided treatment methods. The subject may be a human, primate, dog, cat, cow, horse, mouse, etc. A subject also refers to an animal used in laboratory testing.

[0098] As used herein, "nanoparticle" refers to a particle composed of one or more polymers, the size of which, in nanometers (nm), includes a linear dimension range of 10 nanometers to 2000 nanometers. As used herein, "linear dimension" refers to the distance between any two points on the surface of a nanoparticle measured in a straight line. Nanoparticles of the present disclosure can be irregular, oblong, spindle-shaped, rod-shaped, cylindrical, pancake-shaped, disc-shaped, spherical, biconcave, or erythrocyte-like. Linear dimensions can be measured using several methods, including, but not limited to, transmission electron microscopy or tunable resistive pulse sensing, which are some of the standard means of measuring nanoparticle size. One widely used technique for measuring nanoparticle size is dynamic light scattering (DLS), which can provide the diameter and polydispersity of nanoparticles. DLS assumes that nanoparticles are essentially spherical and that the size of a nanoparticle is the average diameter (or radius) of such an assumed sphere. In such measurements, nanoparticles can be described as having a size range of 10 nm to 1000 nm or 1 nm to 500 nm.

[0099] As used herein, "microparticle" refers to a small particle composed of one or more polymers whose size in micrometers (μm) is less than 1000 μm and includes a maximum cross-sectional width of 1 μm or more.

[0100] As used herein, "polymer" refers to a molecule composed of multiple repeating structural units linked by covalent bonds. As used herein, "polymer particle" refers to a solid or porous particle, in contrast to the shell-like structures of liposomes and polymersomes and the relatively open structure of hydrogel particles. As used herein, "hydrogel particle" refers to a crosslinked network of polymer chains that is absorbent yet stable in an aqueous environment.

[0101] As used herein, the term "biocompatible polymer" refers to a polymer that does not undesirably interfere with the biological function of tissue. Biocompatible polymers may, in some cases, be biodegradable, bioabsorbable, and bioerodible. Biodegradable, bioabsorbable, and bioerodible, as well as degraded, eroded, and absorbed, are used interchangeably (unless the context indicates otherwise) and refer to polymers and materials that can be broken down or absorbed when exposed to body fluids, such as blood, and their components, such as enzymes, and can be gradually resorbed, absorbed, and / or eliminated by the body.

[0102] The terms "treatment" or "treating" are used herein to characterize methods or processes that aim to (1) delay or prevent the onset of a disease, disorder, or condition; (2) slow or halt the progression, worsening, or deterioration of one or more symptoms of a disease, disorder, or condition; (3) bring about an improvement in the symptoms of a disease, disorder, or condition; (4) reduce the severity or incidence of a disease, disorder, or condition; or (5) cure a disease, disorder, or condition. A treatment can be administered prior to the onset of a disease, disorder, or condition for a prophylactic or preventative effect. Alternatively, or in addition, a treatment can be administered after the initiation of a disease, disorder, or condition for a therapeutic effect.

[0103] As used herein, "effective amount" refers to an amount of an active compound agent that induces a desired biological response in a subject, e.g., an amount that results in treatment as defined herein above. In one embodiment, an effective amount of a compound of the invention is from about 0.01 mg / kg / day to about 1000 mg / kg / day, from about 0.1 mg / kg / day to about 100 mg / kg / day, or from about 0.5 mg / kg / day to about 50 mg / kg / day.

[0104] particle In one aspect, the present disclosure provides nanoparticles comprising a polymer that provides for tethering via covalent chemical conjugation of a ligand (e.g., a compound represented by structural formula S-1, S-2, D-1, D-2, S-1-1, S-2-1, D-1-1, D-2-1 or a moiety represented by S-1A, S-2A, D-1A, D-2A, also referred to herein as a sialic acid-containing ligand) for presentation on the nanoparticle surface. The nanoparticles can be used to contact immune cells expressing sialic acid-binding immunoglobulin-like lectins (Siglecs) to modulate inflammatory processes. It has been determined that providing a ligand capable of targeting and binding to immune cells expressing sialic acid-binding immunoglobulin-like lectins (Siglecs) can be used to modulate inflammatory responses in the targeted cells and associated environments.

[0105] Displaying a ligand on the surface of a nanoparticle means that the ligand is decorated on the nanoparticle so that it is available to be bound by a Siglec receptor on a target cell or organism. Suitably, they may be provided to bind, activate or block the receptor.

[0106] A single nanoparticle can be decorated with multivalent ligands, thereby allowing for multivalent binding of different Siglec receptors by this single nanoparticle, resulting in modulation of the inflammatory response. Nanoparticles decorated with unique ligands can also be mixed with other ligand-decorated nanoparticles that can target different Siglec receptors, again allowing for desired modulation of the inflammatory response. In some embodiments, the presentation of a ligand on the surface of a nanoparticle or microparticle can provide an increase in cellular uptake of the particle by at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, or at least about 10-fold. In some embodiments, the presentation of a ligand on a nanoparticle or microparticle can reduce the inflammatory response. In non-limiting embodiments, the reduction in the inflammatory response is greater than about 2-fold, greater than about 3-fold, greater than about 4-fold, greater than about 5-fold, greater than about 10-fold, greater than about 20-fold, greater than about 50-fold, greater than about 100-fold, greater than about 500-fold, or greater than about 1000-fold.

[0107] Nanoparticles or microparticles can be used for systemic or local delivery to target diseased tissues in a subject in need of treatment, resulting in modulation of the inflammatory response in the subject, resolving innate and adaptive inflammation, activating innate and / or adaptive immunity when enhanced immune surveillance is desired, or reducing the infectivity of infectious organisms. The targeted immune cells or viruses should have Siglec receptors or viral sialic acid-ligand binding regions, respectively. Innate immune system activity includes, for example, the cellular response of the innate immune system; the non-cellular / humoral response of the innate immune system; the complement system; the alternative complement pathway; the amplification loop of the alternative complement pathway; and / or the amplification loop of the alternative complement pathway activated by complement factor H. Adaptive immune system activity includes dendritic cell maturation and presentation to T cells, T cell activation, T cell regulation, T cell checkpoint inhibition or activation, neutrophil NETosis, and B cell activation. Reducing infectivity includes reducing viral entry into host cells, reducing viral particle reproduction, or reducing the inflammatory response to viral infection.

[0108] Several types and configurations of nanoparticles are encompassed by the present disclosure. For example, nanoparticles can be composed of a variety of materials, including, but not limited to, biodegradable polymers, biocompatible polymers, bioabsorbable polymers, or combinations thereof.

[0109] The polymer backbone of the nanoparticles onto which the sialic acid-containing ligands are attached can be composed of naturally occurring polymers such as carbohydrates or proteins, or can be composed of synthetic polymers. The polymer backbone will have unique terminal functional groups to provide tethering of the sialic acid-containing ligands to the nanoparticle surface. The polymer backbone can be first attached to multiple sialic acid ligands before forming the nanoparticles via chemical conjugation methods, or the polymer backbone can be first formed into nanoparticles, and then functional groups displayed on the surface of the nanoparticles can be attached to the sialic acid-containing ligands via chemical conjugation methods. Suitable nanoparticles include polymer particles and hydrogel particles.

[0110] Polymers suitable for preparing nanoparticles include, but are not limited to, poly(lactide-co-glycolide)-poly(ethylene glycol) or poly(lactide-co-glycolide)-block-poly(ethylene glycol) or poly(lactide-co-glycolide) containing click chemistry functional groups (e.g., azides or alkynes, such as DBCO) or amine-reactive esters such as succinimidyl esters. Such polymers include, but are not limited to, PLGA-PEG-DBCO and PLGA-PEG-NHS. Other polymers include poly(N-acetylglucosamine) (chitin), chitosan, poly(3-hydroxyvalerate), poly(lactide-co-glycolide (e.g., poly(D,L-lactide-co-glycolide), poly(L-lactide-co-glycolide)), poly(3-hydroxybutyrate), poly(4-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate), polyorthoesters, polyanhydrides, poly(glycolic acid), poly(glycolide), poly(lactic acid) (e.g., poly(L-lactic acid), poly(D,L-lactic acid)), poly(lactide) (e.g., poly(D,L-lactide)), poly(D,L-lactide)-b-poly(ethylene glycol)-azide , poly(D,L-lactide)-b-poly(ethylene glycol)-methyltetrazine, poly(L-lactide-co-D,L-lactide), poly(D,L-lactide)-b-poly(ethylene glycol)-b-poly(ethylene glycol)-carboxylic acid, poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolide), poly(lactide-co-glycolide)-b-poly(ethylene glycol)-b-poly(lactide-co-glycolide), poly(lactide-co-glycolide)-b-poly(ethylene glycol)-azide, poly(lactide-co-glycolide)-b-poly(ethylene glycol)-alkyne, poly((D,L)lactic acid)-b-poly(ethylene glycol)-azide, poly((D,L)lactic acid)-b-poly(ethylene glycol)-alkyne, poly(caprolactone), poly(caprolactone)-b-poly(ethylene glycol), polycaprolactone-b-poly(ethylene glycol), poly(lactide-co-caprolactone)-b-poly(ethylene glycol)-b-poly(lactide-co-caprolactone), poly(L-lactide-co-caprolactone), poly(L-lactide-co-caprolactone), poly(D,Poly(L-lactide-co-caprolactone), poly(glycolide-co-caprolactone), poly(DL-lactide)-b-poly(ethylene glycol)-b-poly(DL-lactide), poly(trimethylene carbonate), polyesteramides, poly(glycolic acid-co-trimethylene carbonate), acrylate-poly(caprolactone)-b-poly(ethylene glycol)-alkyne, co-poly(ether esters) (e.g., PEO / PLA), poly(N-isopropylacrylamide-co-acrylic acid), poly(N-isopropylacrylamide-co-methoxypoly(ethylene glycol) methacrylate), polyphosphazenes, biomolecules (such as fibrin, fibrin glue, fibrinogen, cellulose, starch, collagen and hyaluronic acid, elastin and hyaluronic acid), polyurethanes, silicones, polyesters, polyolefins, polyisobutylene and ethylene-alpha-olefin copolymers, acrylics other than polyacrylates vinyl polymers and copolymers, vinyl halide polymers and copolymers (such as polyvinyl chloride), polyvinyl ethers (such as polyvinyl methyl ether), polyvinylidene halides (such as polyvinylidene chloride), poly(vinylidene fluoride), poly(vinylidene fluoride-co-hexafluoropropylene), polyacrylonitrile, polyvinyl ketone, polyvinyl aromatic compounds (such as polystyrene), polyvinyl esters (such as polyvinyl acetate), acrylonitrile-styrene copolymers, ABS resins, polyamides (such as nylon 66 and polycaprolactam), polycarbonates such as tyrosine-based polycarbonates, polyoxymethylene, polyimides, polyethers, polyurethanes, rayon, rayon-triacetate, cellulose, cellulose acetate, cellulose butyrate, cellulose acetate butyrate, cellophane, cellulose nitrate, cellulose propionate, cellulose ethers, carboxymethyl cellulose, and fullerenes.

[0111] In one aspect, the nanoparticles are formed from polycaprolactone, and in other embodiments, from polymers including polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, and poly(3-hydroxybutyric acid). In some embodiments, the nanoparticles may be formed from poly(lactide) (PLA), poly(glycolide) (PGA), polylactic-glycolic acid (PLGA), poly(butyl cyanoacrylate) (PBCA), or N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer. In other aspects, the nanoparticles are formed from polymers such as poly(ethylene glycol), polyethylene oxide, Pluronic F127, Pluronic F68, poloxamer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, and poly(vinylpyrrolidone).

[0112] In some embodiments, the nanoparticles are formed from a mixture of biodegradable and non-biodegradable polymers as block copolymers (BCPs), including the preferred embodiment of PLGA-block-PEG. Block copolymers include polymers having two or more different polymer subunits covalently linked. In some embodiments, the nanoparticles are formed from a mixture of biodegradable and non-biodegradable polymers as block copolymers, including the preferred embodiment of PLGA-block-PEG. In yet another aspect, the nanoparticles are formed from naturally occurring polymers in the form of hydrogel nanoparticles, including those formed from collagen, hyaluronic acid, heparin, heparin sulfate, chitosan, and alginate.

[0113] Methods for synthesizing nanoparticles are well known to those skilled in the art (see, e.g., Spence et al., Science Translational Medicine, 2015, 7:303 303ra140 and references cited therein). For example, methods for synthesizing nanoparticles with known decomposition rates are known to those skilled in the art and are described in U.S. Patent No. 6,451,338 to Gregoriadis et al., U.S. Patent No. 6,168,804 to Samuel et al., and U.S. Patent No. 6,258,378 to Schneider et al., which are incorporated herein by reference in their entireties.

[0114] Chemical attachment of the ligands described herein to the nanoparticle surface can be achieved through the use of click chemistry reactions. In such reactions, a chemical reaction occurs between a terminal functional group of the nanoparticle polymer and a terminal functional group of the ligand (referred to herein as a "terminal functional conjugate pair"), resulting in the attachment of the polymer to the sialic acid-containing ligand. The type of terminal functional group present on the surface of the polymer and its binding partner ligand determines the type of click chemistry reaction used to chemically attach the ligand to the nanoparticle surface. Furthermore, the selection of a polymer with a specific terminal functional group can be used to control the type, density, and spatial arrangement of the ligand conjugate partners to be presented on the surface of the nanoparticle. In some embodiments, the polymers have specific functional groups that provide chemical binding sites on the surface of the formed nanoparticles, including azides, alkynes, aryl esters, amides, amines, arylamides, aldehydes, acetyls, substituted aryl esters, alkyl esters, alkyl ketones, aryl ketones, substituted aryl ketones, ketones, alkyl halides, aminooxys, alcohols, aza-ylides, carboxylic acids, esters, amides, bicyclononynes, dihydrazides, halo-carbonyls, halosulfonyls, hydrazides, N -hydroxysuccinimide, succinimidyl ester, mono- and difluorinated cyclooctynes, isothiocyanates, iodoacetamides, maleimides, methylcyclopropenes, hydrazines, nitriles, nitros, phosphines, phosphazides, tertazines, methyltetrazines, trans-cyclooctene, strained alkynes, dibenzocyclooctynes, biarylazacyclooctynones, azadibenzylcyclooctynes, vinyls, sulfonyl esters, thioesters, thiocarboxylates, thioesters, sulfonyl halides, thiols, and thiolenes. In certain embodiments, the polymer is poly(lactide-co-glycolide)-poly(ethylene glycol) or poly(lactide-co-glycolide)-block-poly(ethylene glycol) or poly(lactide-co-glycolide) having at least one of the functional groups listed immediately above.

[0115] Such conjugation sites provide locations for the attachment of ligands to the surface of the nanoparticles through the performance of click chemistry reactions. In one example, nanoparticles are formed from PLGA-PEG polymers with azide or alkyne terminal functional groups. In non-limiting embodiments, blends of various polymers with different terminal functional groups can be used. Such polymers include, for example, PLGA-PEG-alkyne, PLGA-PEG-ester, and PLGA-PEG-DBCO. In certain aspects, blends of PLGA-PEG-alkyne and PLGA-PEG-carboxylic acid can be prepared as nanoparticles. In another specific aspect, blends of PLGA-PEG-alkyne and PLGA-PEG-ester can be prepared as nanoparticles. In another specific embodiment, blends of PLGA-PEG-DBCO and PLGA-PEG-carboxylic acid can be prepared as nanoparticles. In another specific aspect, blends of PLGA-PEG-DBCO and PLGA-PEG-ester can be prepared as nanoparticles.

[0116] Although PLGA polymers can have free terminal alkyne groups, many of these are buried in the particle matrix and unavailable for attachment to the particle surface. In some embodiments, more alkyne groups can be introduced to the particles by adding a second polymer or copolymer surfactant or coating to the particle's first PGLA polymer or copolymer. Preferably, the second polymer or copolymer can be branched or linear and can contain multiple terminal alkyl groups, where the alkyl groups contain only carbon and hydrogen, forming a homologous series with the general formula CnH2n+1. In other embodiments, sialic acid-containing ligands can be attached to particles, such as polymeric nanoparticles, via covalent bonds.

[0117] In other embodiments, the ligand comprises a terminal functional group (i.e., a conjugation site) that achieves anchoring to the nanoparticle surface, such as azide, alkyne, aryl ester, amide, amine, arylamide, aldehyde, acetyl, substituted aryl ester, alkyl ester, alkyl ketone, aryl ketone, substituted aryl ketone, ketone, alkyl halide, aminooxy, alcohol, aza-lyde, carboxylic acid, ester, amide, bicyclononyne, dihydrazide, halocarbonyl, halosulfonyl, hydrazide, N-hydroxysuccinimide, norbornene, oxanorbornadiene, succinimidyl ester, isothiocyanate, or the like. Examples of conjugation moieties include acetamides, iodoacetamides, mono- and difluorinated cyclooctynes, maleimides, methylcyclopropenes, isocyanopropanoates, hydrazines, nitriles, nitros, phosphines, phosphazides, tertazines, methyltetrazines, trans-cyclooctene, strained alkynes, dibenzocyclooctynes, biarylazacyclooctynones, propargyls, isocyanides, azadibenzylcyclooctynes, vinyls, sulfonyl esters, thioesters, thiocarboxylates, thioesters, sulfonyl halides, thiols, and thiolenes. Such conjugation moieties provide a location for the attachment of ligands to the surface of nanoparticles through the performance of click chemistry reactions.

[0118] In one embodiment, click chemistry reactions are used to attach ligands to nanoparticle surfaces. These click chemistry reactions are characterized as a class of biocompatible small molecule reactions commonly used in bioconjugation reactions and chemical ligation to modify other molecules, biomolecules, nanoparticles, and other surfaces. In general, click chemistry reactions possess the following properties: modularity, insensitivity to solvent parameters, high chemical yield, insensitivity to oxygen and water, regiospecificity and stereospecificity, and a large thermodynamic driving force (>20 kcal / mol) that favors reactions with a single reaction product. Click chemistry reactions offer high reaction specificity and confer control over both regiospecificity and stereospecificity. This reaction specificity is particularly useful in achieving the desired display of sialic acid ligands on the nanoparticle surface, thereby enabling optimal binding of the nanoparticles to immune cell Siglec receptors. The bond formed by the click reaction during conjugation provides access to an extremely stable covalent bond between the sialic acid ligand and the nanoparticle, which does not undergo rearrangement or reaction or degradation or hydrolysis under biological conditions.

[0119] A variety of different click chemistry reactions can be used to link ligands to the surface of nanoparticles. The use of such click chemistry reactions provides a controlled reaction medium for producing nanoparticles with desired sialic acid ligand density and spatial arrangement. The density and spatial arrangement of sialic acid ligands on the nanoparticle surface can be controlled, for example, by adjusting the amount of functionalized polymer used to form the nanoparticles, polymer molecular weight, polymer density, the number of functional groups per polymer, solvent, type of functional group, concentration of ligand, type of click chemistry used, and type of click chemistry conjugate pair.

[0120] In various embodiments, the average molecular weight of a polymer, e.g., PEG, PLGA, PEG-PLGA block copolymer, can be determined by any method known in the art, such as anion exchange chromatography, gel permeation chromatography, viscosity measurement, among others.

[0121] Click chemistry reactions used to tether ligands to polymers are well known to those skilled in the art and include, for example, Huisgen 1,3-dipolar cycloaddition, copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to produce 1,3-substituted products, ruthenium-catalyzed alkyne-azide cycloaddition (RuAAC) to produce 1,5-substituted triazoles, strain-promoted alkyne azide cycloaddition (SPAAC) to produce 1,4-substituted products, strain-promoted alkyne-nitrone cycloaddition, alkene and tetrazine reverse demand Diels-Alder, tetrazine trans-cyclooctene ligation, thiol-ene reaction, thiol-yne reaction, Staudinger reaction, [4+1] cycloaddition, quadricyclane ligation [2+2+2] cycloaddition, norbornene cycloaddition, and alkene-tetrazole photoclick reaction.

[0122] The tethering of the ligand to the surface of the nanoparticle is performed in a manner that achieves presentation of the ligand for maximum binding affinity to Siglec receptors expressed on the surface of immune cells or sialic acid ligand receptors expressed on the surface of viral particles. The ligand density can be adjusted to provide a desired multivalent or polyvalent ligand interaction with the Siglec receptor upon contact with immune cells, as such an interaction correlates with a desired cellular immune response. The multivalent or polyvalent ligand-receptor interaction can be controlled based on the density of the ligand provided on the nanoparticle surface, which can affect the response elicited by the immune cell upon contact.

[0123] As disclosed, click chemistry reactions require functionalization of both the nanoparticle polymer and the sialic acid ligand to enable the desired attachment of the ligand to the nanoparticle surface. In one embodiment, a terminal alkyne is presented on the nanoparticle surface for covalent chemical conjugation with the azide-presented sialic acid ligand through copper(I)-catalyzed azide-alkyne (CuAAC) reaction; copper-free reaction; strain-promoted azide-alkyne reaction (SPAAC); tetrazine-alkene ligation reaction; or trans-cyclooctene (TCO)-tetrazine reaction. For example, azide functional groups can be added to the ligand using sialyltransferase ST8SIA4.

[0124] In one embodiment, ligand / nanoparticle conjugate pairs can be prepared using copper(I) azide-alkyne cycloaddition of azide-bearing ligands with alkyne-functionalized nanoparticles or alkyne-bearing ligands with azide-functionalized nanoparticles. Ligands containing dibenzylcyclooctyne, difluorooctyne, or biarylazacyclooctynone can be reacted with azides via SPACC. Ligands containing transcyclooctene can be reacted with tetrazine-functionalized nanoparticles.

[0125] Nanoparticles can be prepared by mixing poly(D,L-lactide-co-glycolide-COOH)-PEG-COOH (PLGA 10,000 Da - PEG-COOH 5,000 Da) with poly(lactide-co-glycolide)-b-poly(ethylene glycol)-alkyne (PLGA-PEG-alkyne; 10,000 Da PLGA:1,000 Da PEG) in a PLGA-PEG-COOH:PLGA-alkyne (DBCO) ratio of 75:25 (w / w). This 75:25 ratio represents one embodiment of the density of alkyne functional groups on the nanoparticle surface. Other ratios of PLGA-PEG-COOH to PLGA-PEG-alkyne (DBCO) used in nanoparticle preparation include 95:5, 90:10, 85:15, 80:20, 70:30, 65:35, 60:40, 55:45, and 50:50. The ratio of PLGA-PEG-COOH to PLGA-PEG-alkyne (DBCO) was designed to provide sufficient space between functional groups to allow efficient conjugation of polymer ligands and achieve the desired ligand density.

[0126] Nanoparticles can be prepared that contain one or more polymers with different click chemistry functional groups for pairing with their respective sialic acid-containing ligand conjugation partners, thereby allowing for the presentation of one or more ligands on the nanoparticle surface at different densities and / or spatial arrangements. By using more than one nanoparticle polymer / ligand pair, nanoparticles can be engineered to use different conjugate pairs.

[0127] By providing different click chemistry functional groups available on the nanoparticle surface, different types of ligands can be conjugated to the nanoparticle surface. The density of the different functional groups can be controlled by the ratio of different polymers, the concentration of the polymers and the type of click chemistry conjugate pair, the type of click chemistry reaction, and the size and shape of the sialic acid-containing ligand. The number of different ligands that can be displayed on the surface can be determined by one of ordinary skill in the art. In a non-limiting embodiment, the number of different ligands present on the nanoparticle surface ranges from 1 to 20. The number of different ligands can be, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In another embodiment, the number of different ligands present on the nanoparticle surface ranges from 2 to 20. The number of different ligands can be, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In another embodiment, the nanoparticles will comprise at least two different ligands. In another embodiment, the nanoparticles will comprise at least three different ligands. In another embodiment, the nanoparticles will comprise at least four different ligands. In another embodiment, the nanoparticles will comprise at least five different ligands.

[0128] Generally, the density of functional groups on the nanoparticle surface determines the maximum ligand density that can be tethered to the nanoparticle surface via covalent chemical bonds via click chemistry. The ligand density can be controlled and quantified in terms of the number of functional groups per square nanometer of surface area.

[0129] The density of the ligands on the nanoparticle surface can be controlled by several methods, including chemical conjugation techniques, ligand density on the polymer, type of ligand, solvent, pH, and ionic strength. The ligand density on the surface of the nanoparticles can be tailored so that contact of immune cells with such nanoparticles results in an immunomodulatory response, including an anti-inflammatory biological response. Control of the ligand density can also be used to modulate the magnitude of the desired anti-inflammatory response.

[0130] In some embodiments, the sialic acid-containing ligands can be displayed on the nanoparticle in groups of at least 2, at least 5, at least 10, at least 15, at least 20, or at least 25, at least 50, at least 100, at least 200, or at least 400. In some embodiments, the sialic acid-containing ligands can be spaced apart on the surface of the nanoparticle so that they or the nanoparticle can bind to more than one Siglec receptor. In some embodiments, the sialic acid-containing ligands can be spaced apart on the surface of the nanoparticle so that they or the nanoparticle can bind to multiple Siglec receptors displayed on individual cell types, which may have varying amounts of Siglec receptors displayed on their plasma membranes.

[0131] In some embodiments, nanoparticles can comprise a polymer containing a sialic acid-containing ligand at a concentration of 0.05 nmol / mg of sialic acid-containing ligand to 250 nmol / mg of sialic acid-containing ligand to nanoparticle, preferably 0.5 nmol / mg to 25 nmol / mg, and most preferably 0.5 to 15 nmol of sialic acid-containing ligand per mg of nanoparticle. In embodiments, devices can be coated with such nanoparticles. In alternative embodiments, devices can be formed from polymers, e.g., the device is a microparticle or nanoparticle, and the sialic acid-containing ligand is provided in the polymer at a concentration of 0.05 nmol / mg of sialic acid-containing ligand to 250 nmol / mg of sialic acid-containing ligand to nanoparticle, preferably 1 nmol / mg to 25 nmol / mg, and most preferably 2 to 15 nmol of sialic acid-containing ligand per mg of nanoparticle.

[0132] In some embodiments, nanoparticles can have a maximum cross-sectional width or diameter of less than about 1000 nm, less than about 500 nm, less than about 250 nm, or less than about 200 nm. In embodiments, nanoparticles can have a width greater than about 1 nm, greater than about 10 nm, greater than about 50 nm, or greater than about 100 nm. In embodiments, nanoparticles coated with sialic acid or sialic acid analogs can have a maximum cross-sectional width or diameter ranging from about 130 nm to about 170 nm, more preferably a width of about 150 nm. In some embodiments, these size ranges can be the average width of the nanoparticles. In some embodiments, at least 80% of the nanoparticles are within the disclosed ranges.

[0133] Preferably, in some embodiments, at least about 80%, more preferably at least 90%, of the particles have a maximum cross-sectional width of 130 nm to 170 nm. In some embodiments, the particles can have an average maximum cross-sectional width of 150 nm, with no particles having a width greater than or less than one standard deviation of 150 nm. In some embodiments, the nanoparticles can have a volume equivalent to a sphere having a diameter of 10 nm to 500 nm, preferably 50 nm to 250 nm, 100 nm to 200 nm, or 130 nm to 170 nm.

[0134] In one example, a nanoparticle may have a volume equivalent to a sphere having a diameter of about 100 nm.

[0135] In another aspect of the present invention, linking nanoparticles to ligands provides a means for nanoparticles to evade the immune system, i.e., opsonization and phagocytosis via the reticuloendothelial system (RES). PEGylation of nanoparticles, i.e., coating nanoparticles with polyethylene glycol, is known to provide a protective barrier from detection by immune cells. However, PEG has drawbacks such as toxicity, immunogenicity, reduced cellular uptake, reduced binding, and non-biodegradable or bioabsorbable properties. Coating nanoparticles with sialic acid-containing ligands overcomes the drawbacks of PEG and provides a natural, non-immunogenic nanoparticle coating that can evade RES and immune detection. Thus, the nanoparticles disclosed herein have the ability to evade immune detection and reduce immunogenic responses.

[0136] The nanoparticles or microparticles disclosed herein can further comprise a bioactive agent encapsulated within the nanoparticle, attached to its surface, or incorporated into its structure. For example, the nanoparticles can further comprise at least one of an antibiotic, an antiviral agent, an anti-inflammatory agent, a cytokine, a cytokine inhibitor, an immunomodulator, an immunotoxin, an anti-angiogenic agent, an antihypertensive agent, an anti-edema agent, a radiosensitizer, an oligonucleotide including DNA or RNA, a peptide, an anticancer agent, or any combination thereof. Methods for preparing nanoparticles containing a bioactive agent encapsulated within the nanoparticle, attached to its surface, or incorporated into its structure are known to those skilled in the art.

[0137] Modified Oligosaccharides Disclosed herein are modified oligosaccharides, generally referred to as S-series (i.e., S-1XX and S-2XX) compounds and D-series (i.e., D-1XX and D-2XX) compounds, also referred to herein as sialic acid-containing ligands. The S-series compounds are compounds represented by the following general structural formula (S-1) (i.e., S-1XX compounds) or (S-2) (i.e., S-2XX compounds), or pharmaceutically acceptable salts thereof. The D-series compounds are compounds represented by the general structural formula (D-1) (i.e., D-1XX compounds) or (D-2) (i.e., D-2XX compounds), or pharmaceutically acceptable salts thereof. [ka]

[0138] Values ​​and example values ​​of the variables R and R1 in formulas (S-1), (S-2), (D-1) and (D-2) are defined herein below.

[0139] Exemplary compounds of the S- and D-series and their syntheses are described in Examples 1A, 1B, and 2 below.

[0140] Conjugation of oligosaccharides to polymers and formation of nanoparticles General structural formula GLP Also described herein are nanoparticles comprising a polymer conjugated to the above-described oligosaccharide, having the formula: [ka] or a pharmaceutically acceptable salt thereof. Values ​​and example values ​​for the variables R, R1, and L in formulas (S-1A), (S-2A), (D-1A), and (D-2A) are described herein below.

[0141] Exemplary compounds of conjugated oligoglycans of formula (S-1A), (S-2A), (D-1A), and (D-2A) and their synthesis are described in Example 3 below.

[0142] Indications In various example embodiments, the compounds and particles described herein are useful in methods for treating diseases and disorders that respond to modulation (e.g., amplification, reduction, or elimination) of Siglec receptor activity, including cancer, immune-related, and inflammatory-related diseases and disorders, including, but not limited to, breast cancer, non-small cell lung cancer (NSCLC), prostate cancer, colorectal cancer, melanoma, pancreatic cancer, and myelofibrosis, diabetic retinopathy, idiopathic pulmonary fibrosis, hepatic fibrosis, sickle cell anemia, and acute respiratory distress syndrome (ARDS).

[0143] In further embodiments, the present disclosure provides methods of treating immune-related and inflammation-related diseases including, but not limited to, dry and wet macular degeneration, retinal vascular disease, diabetic retinopathy, diabetic macular edema, cystoid macular edema, proliferative diabetic retinopathy, proliferative vitreoretinopathy, dry eye, allergic conjunctivitis, rheumatoid arthritis, arthritis, lupus, nephritis, immune complex nephropathy, allergic esophagitis, allergic gastritis, hepatitis, liver fibrosis, idiopathic pulmonary fibrosis, acute respiratory distress syndrome, sepsis, bacterial and viral infections, influenza, SARS-CoV-1 and SARS-CoV-2, HIV / AIDS, group B streptococcal infections, Neisseria infections, cancers involving solid organs, non-small cell lung cancer, colorectal cancer, prostate cancer, uveal melanoma, malignant melanoma-skin, myelofibrosis or hematopoietic cancers, in each case in an afflicted subject through administration of such pharmaceutical compositions. The present disclosure provides methods of modulating an inflammatory response in a cell, the method comprising providing a cell with a sialic acid-containing ligand, the sialic acid-containing ligand being presented on a nanoparticle such that a pro-inflammatory response in the cell is suppressed or an anti-inflammatory response in the cell is increased. In certain embodiments, the method provides for suppression of a pro-inflammatory response. In alternative embodiments, the method provides for an increase in an anti-inflammatory response. In some embodiments, the method provides for an enhancement of a pro-inflammatory response in conditions such as infection or cancer.

[0144] Thus, there is provided a method of treating an inflammatory disease in a subject in need thereof, said method comprising administering sialic acid or an analog thereof to the subject, wherein the sialic acid-containing ligand is presented on a nanoparticle such that a pro-inflammatory immune response is suppressed or an anti-inflammatory immune response is increased in the subject.

[0145] The method may include identifying a subject having a proinflammatory immune response and / or suffering from a disorder associated with or caused by a proinflammatory immune response, or at risk of developing a proinflammatory immune response or a disorder associated with or caused by a proinflammatory immune response; administering to the subject a sialic acid-containing ligand, wherein the sialic acid-containing ligand is presented on a nanoparticle.

[0146] In specific embodiments, the methods can be used to treat a subject having a pulmonary disease, including inflammatory and non-inflammatory diseases of the lung, including, but not limited to, tuberculosis, chronic obstructive pulmonary disorder (COPD), asthma, acute lung injury, acute respiratory distress syndrome, cystic fibrosis, bronchiectasis, pulmonary fibrosis interstitial lung disease, pulmonary vascular disease, influenza, viral pneumonia, bacterial pneumonia, allergic bronchitis, non-allergic bronchitis, rhinitis, and fibrosing alveolitis.

[0147] In some embodiments, the methods can be used to treat rheumatic diseases, including but not limited to rheumatoid arthritis, fibromyalgia, systemic lupus erythematosus, systemic sclerosis (scleroderma), psoriatic arthritis, ankylosing spondylitis, Sjogren's syndrome, polymyalgia rheumatica, gout, osteoarthritis, infectious arthritis, and juvenile idiopathic arthritis.

[0148] In some embodiments, the methods can be used to treat gastrointestinal inflammation, including but not limited to Crohn's disease, ulcerative colitis, irritable bowel syndrome, celiac disease, diverticulitis, gastroesophageal reflux disease, lactose intolerance, peptic ulcer, cholecystitis, gastritis, colitis, pancreatitis, autoimmune hepatitis, hepatitis, infectious hepatitis, and pancreatitis.

[0149] In some embodiments, the methods can be used to treat cardiovascular diseases, including but not limited to, septic shock, atherosclerosis, diastolic dysfunction, heart failure, cardiac fibrosis, Coxsackie myocarditis, congenital heart block, autoimmune myocarditis, giant cell myocarditis, and inflammation.

[0150] In some embodiments, the method can be used to treat renal inflammation, including but not limited to, kidney transplant rejection, glomerulonephritis, acute nephritis, cystitis, prostatitis, diabetic nephritis, diabetic kidney disease, and urinary tract infections.

[0151] In some embodiments, the method can be used to treat dermatological inflammation, including but not limited to dermatitis, eczema, inflammatory rash, scleroderma, keloids, acne, sarcoidosis, tinea cruris, tinea corporis, tinea pedis, tinea capitis, tinea unguium, rosacea, vitiligo, lichen sclerosis, autoimmune urticaria, dermatomyositis, and hidradenitis suppurativa.

[0152] In some embodiments, the methods can be used to treat neurological inflammation and degeneration, including but not limited to neuromyelitis, multiple sclerosis, encephalitis, neurosarcoidosis, Alzheimer's, amyotrophic lateral sclerosis, and Huntington's disease.

[0153] In some embodiments, the methods can be used to treat autoimmune inflammation, including, but not limited to, diabetes, SLE, multiple sclerosis, Sjogren's syndrome, Addison's disease, Graves' disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, celiac disease, pernicious anemia, alopecia areata, autoimmune hepatitis, autoimmune angioedema, autoimmune encephalomyelitis, autoimmune inner ear disease, Guillain-Barré, Kawasaki disease, Lambert-Eaton syndrome, Vogt-Koyanagi-Harada syndrome, systemic vasculitis, giant cell arteritis, sarcoidosis, and polyarteritis nodosa.

[0154] In some embodiments, the methods can be used to treat viral inflammation, including but not limited to, influenza A, B, C, SARS-CoV1, SARS-CoV2, Newcastle Disease, Sendai virus, Polyomavirus, HIV, Flavivirus, Caclivirus, Herpes virus, Picoronovirus, and Coronavirus.

[0155] In some embodiments, the method can be used to treat fungal inflammation, including but not limited to fungemia, fungal abscesses, fungal keratitis, candidiasis, tinea pedis, and tinea cruris.

[0156] In some embodiments, the methods can be used to treat parasitic infections, including but not limited to amebiasis, giardiasis, toxoplasmosis, and toxocara.

[0157] In some embodiments, the methods can be used to treat fibrosis, including idiopathic pulmonary fibrosis, bone marrow fibrosis, liver fibrosis, cardiac fibrosis associated with diastolic dysfunction and CHF, renal fibrosis, retinal fibrosis, skin fibrosis, and scarring.

[0158] In some embodiments, the methods can be used to treat acute, life-threatening inflammation, including, but not limited to, sepsis and cytokine storm. In specific embodiments, methods are provided for treating multiple ocular inflammatory diseases, such as macular degeneration, uveitis, optic neuritis, neuromyelitis, and inflammation resulting from ocular infections, drug and toxin exposure, as well as general immune disorders, including autoimmune diseases. In non-limiting embodiments, methods are provided that are useful for preventing, treating, or ameliorating macular degeneration, such as dry macular degeneration, wet macular degeneration, geographic atrophy, moderate macular degeneration, and age-related macular degeneration, in patients. The methods for treating, preventing, or ameliorating ocular inflammation, including macular degeneration, comprise administering a composition of sialic acid-ligand nanoparticles to a patient suffering from or at risk of developing ocular inflammation, such as macular degeneration.

[0159] In some embodiments, the ophthalmic preparation is provided as eye drops, eye ointments, or ophthalmic injections, in which case intravitreal or subconjunctival injections can be used to administer the nanoparticles.

[0160] The co-administration of additional compounds that are used in the method for treating, preventing or improving macular degeneration can be co-administered with the nanoparticle-containing pharmaceutical composition used to treat macular degeneration.For example, anti-angiogenic drugs for treating wet age-related macular degeneration, such as pegaptanib sodium, ranibizumab, bevacizumab, aflibrecept and brolucizumab, can be used as co-administration agents.Although specific embodiments of the present disclosure have been shown and described, it will be clear to those skilled in the art that changes and modifications can be made without departing from the disclosure in its broader aspects.Therefore, the appended claims are intended to encompass within their scope all changes and modifications that fall within the true spirit and scope of the present disclosure.

[0161] In example embodiments, the compounds described herein can be used in methods for treating the following disorders and conditions:

[0162] Ophthalmic diseases such as age-related macular degeneration, dry eye, diabetic retinopathy, allergic ophthalmopathy and ocular fibrosis.

[0163] Neurological diseases such as Alzheimer's "neuroinflammation / degeneration, ALS, multiple sclerosis, and neuropsychiatric "schizophrenia."

[0164] Cancers including solid tumors (breast, lung, colon, prostate, brain, glioblastoma), lymphoma and leukemia.

[0165] Infectious diseases such as viral infections including influenza, avian influenza and SARS-COV-2, bacterial infections such as meningococcal, streptococcal, and group B streptococcus.

[0166] Allergic conditions such as anaphylactic shock, food allergies and contact allergies.

[0167] Fibrosis, including pulmonary fibrosis, hepatitis-induced fibrosis, and bone marrow fibrosis.

[0168] Autoimmune conditions such as uveitis, arthritis, systemic lupus erythematosus, and sepsis.

[0169] Kidney disease such as AHUS or glomerulonephritis.

[0170] GI conditions such as Crohn's disease or ulcerative colitis.

[0171] Cardiac diseases such as atherosclerosis "macrophage-driven", cardiac fibrosis, ischemia-induced cardiomyopathy, and heart failure.

[0172] Inflammatory conditions In other embodiments, the compositions disclosed herein can provide an inhibition of the pro-inflammatory response and an increase in the anti-inflammatory response.

[0173] In one aspect, the pro-inflammatory response can be suppressed by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99%. In alternative embodiments, the anti-inflammatory response can be increased by at least 10%, at least 20%, at least 30%, at least 40%, and at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99%.

[0174] Suitably, pro-inflammatory cytokines can be measured to determine the efficacy of nanoparticle drug treatment. Such measurements can be performed during actual treatment of subjects or, alternatively, during animal testing of the nanoparticles disclosed herein. In embodiments, pro-inflammatory cytokines can include, for example, TNF-α and IL-6. Suitably, anti-inflammatory cytokines, such as IL-10, can also be measured. Those skilled in the art will know suitable assay methods for measuring such cytokines. For example, the Bio-Plex™ Cytokine Assay (Bio-Rad) can be used. A suitable method that can be used to determine whether cells produce more or less pro-inflammatory cytokines is to resuspend the cells and measure 2 x 10 5 The cells are seeded into 96-well plates at 200 μl per well and left overnight to adhere to the plate. They can then be treated with LPS and ligands at a range of concentrations for 24 hours. The supernatant can then be removed and stored at -70°C. Cytokine levels can then be assessed by ELISA (R&D systems). As will be appreciated, similar methods can be applied to determine anti-inflammatory cytokines.

[0175] In an embodiment, TNF-α levels can be suitably determined by coating a 96-well plate overnight with a TNF-α capture antibody diluted in 1x phosphate-buffered saline (PBS). All steps can be performed at room temperature. The wells can be washed three times in 1x PBS / 0.1% polyoxyethylene sorbitan monolaurate (Tween 20) and then blocked with 1% BSA (BDH) in 1x PBS for 1 hour. The washing step can be repeated, and 50 μl of treated cell supernatant or standard ranging from 2000 pg / ml to 0 pg / ml can be added to the wells and left for 2 hours. Subsequently, the supernatant can be aspirated, the wells washed three times, and 50 μl of TNF-α detection antibody diluted in 1% BSA / 1x PBS can be added for 2 hours. The wells can again be washed three times, and horseradish peroxidase (HRP)-conjugated antibody can be added at a 1 in 200 dilution in 1% BSA / 1x PBS for 20 minutes. At this stage, the plate can be covered with aluminum foil. After washing the wells, 3,3',5,5'-tetramethylbenzidine (TMB) can be added for 20 minutes, again protected from light. The reaction can be stopped by adding 1 M hydrochloric acid, and the absorbance can be read at 450 nM on a plate reader. TNF-α concentrations can then be extrapolated from the standard curve. As will be appreciated, similar methodologies can be applied to determine other cytokine levels, substituting the TNF-α detection antibody with other detection antibodies or agents specific for the applicable cytokine.

[0176] In an embodiment, IL-10 levels can be suitably determined by coating a 96-well plate overnight with an IL-10 capture antibody diluted in 1x phosphate-buffered saline (PBS). All steps can be performed at room temperature. The wells can be washed three times in 1x PBS / 0.1% polyoxyethylene sorbitan monolaurate (Tween 20) and then blocked with 1% BSA (BDH) in 1x PBS for 1 hour. The washing step can be repeated, and 50 μl of treated cell supernatant or standard ranging from 2000 pg / ml to 0 pg / ml can be added to the wells and left for 2 hours. Subsequently, the supernatant can be aspirated, the wells washed three times, and 50 μl of IL-10 detection antibody diluted in 1% BSA / 1x PBS can be added for 2 hours. The wells can again be washed three times, and horseradish peroxidase (HRP)-conjugated antibody can be added at a 1 in 200 dilution in 1% BSA / 1x PBS for 20 minutes. At this stage, the plate can be covered with aluminum foil. After washing the wells, 3,3',5,5'-tetramethylbenzidine (TMB) can be added for 20 minutes, again protected from light. The reaction can be stopped by adding 1 M hydrochloric acid, and the absorbance can be read at 450 nM on a plate reader. IL-10 concentrations can then be extrapolated from the standard curve. As will be appreciated, similar methodologies can be applied to determine other cytokine levels, substituting the IL-10 detection antibody with other detection antibodies or agents specific for the applicable cytokine.

[0177] One method that can be used to determine whether a treated subject or test animal develops a higher or lower proinflammatory response is the analysis of serum cytokine levels. For example, this can be accomplished by collecting 50 μl of blood from the treated subject using a capillary tube. The blood is allowed to clot at room temperature for 30 minutes and then centrifuged at 1300 rpm to pellet red blood cells. The serum is decanted into a clean microcentrifuge tube and analyzed by ELISA. For more extensive analysis, larger volumes of blood (approximately 600 μl to 1 ml) can be collected by direct cardiac puncture, thus allowing larger volumes of serum to be collected and analyzed by ELISA or other such techniques. Other suitable techniques for determining whether a treated subject or test animal develops a higher or lower proinflammatory response, particularly for detecting and measuring cytokines, are known in the art.

[0178] Pharmaceutical Composition As used herein, "pharmaceutically acceptable carriers or excipients" can include any and all medically suitable inactive ingredients and / or solvents, diluents or other liquid vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, fillers, extenders, lubricants, etc., appropriate for the particular dosage form desired. Remington's Pharmaceutical Sciences, Eighteenth Edition, A.R. Geunaro (Mack Publishing Co., Easton, Pa., 1990) discloses various carriers used in formulating pharmaceutical compositions and known techniques for their preparation.

[0179] The present disclosure further provides pharmaceutical or veterinary compositions comprising the sialic acid-containing, ligand-conjugated nanoparticles disclosed herein. Such pharmaceutical compositions are formulated to be compatible with their intended route of administration. Examples of routes of administration include both parenteral and non-parenteral methods of administration, including, for example, intravenous, intravitreal, oral, intraocular, subretinal, subtenon, intrascleral, periocular, intravenous, nasal and oral inhalation, intramuscular, intraarterial, intraspinal, intrathecal, intracranial, intradermal, transdermal (topical), transmucosal, subcutaneous, pulmonary lavage, gastric lavage, intrahepatic, subcutaneous, and rectal administration.

[0180] Suitably, in some embodiments, the nanoparticles can be administered parenterally. After parenteral administration, the nanoparticles can selectively accumulate in specific tissues or body sites. In some embodiments, the nanoparticles can deliver a therapeutic payload to cells or tissues. In some embodiments, the nanoparticles can access diseased tissues via enhanced permeability and retention effects.

[0181] Generally, pharmaceutical compositions are provided that contain an effective amount of nanoparticles together with pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants, and / or carriers. Such compositions may contain additives such as diluents of various buffer contents (e.g., Tris-HCl, acetate, phosphate), pH, and ionic strength; detergents and solubilizers (e.g., Tween 80, polysorbate 80), antioxidants (e.g., ascorbic acid, sodium metabisulfite), preservatives (e.g., thimersol, benzyl alcohol), and bulking agents (e.g., lactose, mannitol). Such compositions may affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the nanoparticles. See, for example, Remington's Pharmaceutical Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, Pa. 18042), pp. 1435-1712, incorporated herein by reference. The compositions may be prepared in liquid form or may be formulated into a dry powder, such as a lyophilized form.

[0182] The term "pharmaceutically acceptable salt" also refers to a salt prepared from a compound disclosed herein or any other compound depicted herein that has an acidic functional group, such as a carboxylic acid functional group, and a pharmaceutically acceptable inorganic or organic base.

[0183] Pharmaceutically acceptable salts of the compounds of the present invention are also included. For example, acid salts of compounds of the present invention containing an amine or other basic group can be obtained by reacting the compound with a suitable organic or inorganic acid to provide a pharmaceutically acceptable anionic salt form. Examples of anionic salts include acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, glyceptate, gluconate, glutamate, glycolylarsanilate, hexylresorcinol, hydrobromide, hydrochloride, hydroxynaphthoate ... These include ethate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methylsulfate, mucoate, napsylate, nitrate, pamoate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, diacetate, succinate, sulfate, tannate, tartrate, teoclate, tosylate and triethiodide salts.

[0184] Salts of compounds of the present invention containing a carboxylic acid or other acidic functional group can be prepared by reacting them with a suitable base. Such pharmaceutically acceptable salts can be made with a base that provides a pharmaceutically acceptable cation, including alkali metal salts (especially sodium and potassium), alkaline earth metal salts (especially calcium and magnesium), aluminum salts, and ammonium salts, as well as physiologically acceptable organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N,N'-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, N,N'-bisdehydroabietylamine, glucamine, N-methylglucamine, collidine, quinine, quinoline, and basic amino acids such as lysine and arginine.

[0185] Thus, in a first exemplary embodiment, the present invention provides a compound having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof.

[0186] In a first aspect of the first exemplary embodiment, R1 is, independently at each occurrence, -C(O)-A, where A is C1-C6 alkyl, C6-C 18 Aryl, (C6-C 18)aryl(C1-C3)alkyl, 5- to 18-membered heteroaryl, (5- to 18-membered)heteroaryl(C1-C3)alkyl, C3-C8 cycloalkyl, (C3-C8)cycloalkyl(C1-C3)alkyl, 5- to 8-membered heterocycloalkyl, or (5- to 8-membered)heterocycloalkyl(C1-C3)alkyl; one or two carbon atoms within the alkyl portion of A are each independently optionally replaced with a heteroatom selected from N, O, or S. (Examples of alkyl portions of A containing heteroatoms are CH3-CH2-O-CH2-CH2-, CH3-NH-CH2-, CH3-N(CH3)-CH2-, CH3-S-CH2-CH2-, etc.). Additionally, A may be selected from the group consisting of one to three R 11 groups, and each of the R 11 The group is C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, halogen, C6-C 12 aryl, 5- to 12-membered heteroaryl, cyano, or two groups R 11 together with the atom to which they are attached form a 5- to 7-membered heterocyclyl having 1 to 3 heteroatoms selected from N, O, or S; R 11 are each independently optionally substituted with 1 to 3 substituents selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or cyano; and R is independently selected at each occurrence from -R L -R F and R L is independently expressed as -O-(C1-C 12 ) Alkylenyl-, -O-, -S-, -NR 100 -, -S-(C1~C 12 ) Alkylenyl-, -NR 101 -(C1~C 12 ) Alkylenyl-, -NR 101a -O-(C1~C 12 )Alkylenyl-;-O-(CH2CH2O) m -, -O-(CH2CH2O) k -(CH2CH2)-, -NR 102 -X 100 -(C1~C 12 ) Alkylenyl-, -NR102a -NR 102b -C(O)-(C1~C 12 ) alkylenyl, and R 100 , R 101 , R 101a , R 102 , R 102a and R 102b are each independently H or C1-C3 alkyl, and X 100 is —O— or —NH—, m and k each independently represent an integer of 1 to 12; R F represents independently at each occurrence H, C1-C3 alkyl, -NH2, -NH-Fmoc, -NH-Boc, -NH-CBz, -NH-Troc, -NH-TFA, mono(C1-C3)alkylamino, di(C1-C3 alkyl)amino; -(O)-R 103 (R 103 is -H, -OH or (C1-C3) alkyl; The following structural formula [ka] or a click chemistry reagent.

[0187] In a second aspect of the first exemplary embodiment, the compound has the following structural formula: [ka] or a pharmaceutically acceptable salt thereof. The values ​​and example values ​​of the variables in formulae (S-1-1), (S-2-1), (D-1-1) and (D-2-1) are defined above in relation to the first aspect.

[0188] In a third aspect of the first example embodiment, the click chemistry reagent comprises an azide, a C2-C3 alkyne, a tetrazine, a trans-cyclooctene, or a cyclooctyne. The remainder of the values ​​and example values ​​are defined above in relation to the first and second aspects.

[0189] In a fourth aspect of the first exemplary embodiment, the click chemistry reagent is an azide, a C2-C3 alkyne, or a hydroxyl group having the following structural formula: [ka] (In the formula, R co is hydrogen or halogen; X is absent or O; and R 0 In each case, hydrogen, halogen, C 1~8 Alkyl, C 1~8 Alkoxy, C6-C 12 Aryl, 5-8 membered heteroaryl, C 3~8 cycloalkyl or 3- to 8-membered heterocyclyl; any two or more R 0 The groups, together with the atoms to which they are attached, can optionally form an unsaturated, saturated, or aromatic 5- to 8-membered ring; R td is hydrogen, C 1~8 Alkyl, C 1~8 Alkoxy, C6-C 12 Aryl, 5-8 membered heteroaryl, C 3~8 cycloalkyl or 3- to 8-membered heterocyclyl. The remainder of the values ​​and example values ​​are defined above in relation to the first to third aspects. is a part represented by one of

[0190] In a fifth aspect of the first exemplary embodiment, the click reagent is an azide, a C2-C3 alkyne, or a compound of the following structural formula: [ka] (In the formula, R * is H or methyl, and R # is independently at each occurrence H or C1-C3 alkyl, and R X is —NH—C(O)O—. The remainder of the values ​​and example values ​​are defined above in relation to the first to fourth aspects. It is one of the parts represented by

[0191] In a sixth aspect of the first exemplary embodiment, R is, independently at each occurrence, —O—(CH) x -NH2 or -O-(CH2) x -NH-Fmoc, and x is, independently at each occurrence, an integer from 1 to 10, e.g., x is 5. The remainder of the values ​​and example values ​​are defined above in relation to the first to fifth aspects.

[0192] In an additional embodiment, R is independently at each occurrence -O-(CH) x -N3, and x is independently at each occurrence an integer from 1 to 10, for example, x is 5.

[0193] In a seventh aspect of the first exemplary embodiment, the cycloalkyl portion of moiety A is selected from C-C cycloalkyl; the heterocycloalkyl portion of moiety A is selected from 5-6 membered heterocycloalkyl having one or two heteroatoms selected from N, O, or S; the aryl portion of moiety A is selected from phenyl or naphthalenyl; and the heteroaryl portion of moiety A has the following structural formula: [ka] X is selected from the part represented by one of 1 , X 2 and X 3 are each independently, NR H , O or S; R H is H or C1-C3 alkyl; when present, each R 11 are independently selected from cyano, halogen, phenyl, halophenyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C3 alkoxy, or two groups R 11 together with the atoms to which they are attached form a [1,3]dioxolo group optionally substituted with one or two methyl or ethyl groups. The remainder of the values ​​and example values ​​are defined above in connection with the first through sixth aspects.

[0194] In an eighth aspect of the first exemplary embodiment, R1 is, independently at each occurrence, selected from the following structural formulas: [ka] where the wavy line represents the point of attachment of R1 to the nitrogen atom.

[0195] The remainder of the values ​​and example values ​​are defined above in relation to the first to seventh aspects.

[0196] In a ninth aspect of the first exemplary embodiment, the compound is any one of compounds S-101 to S-131 or D-101 to D-131, and the moiety R is -O-(CH2)5-NH-Fmoc.

[0197] In additional embodiments, the compound is any one of compounds S-101 through S-131 or D-101 through D-131, and the moiety R is —O—(CH 2 ) 5 —NH 2 or —O—(CH 2 ) 5 —N 3 .

[0198] In a second exemplary embodiment, the present invention provides a compound having the following structural formula: GLP The particle includes a compound represented by the formula:

[0199] In a first aspect of the second example embodiment, P is a biocompatible polymer; L is a covalent linker; and G is a group having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof.

[0200] In formulas (S-1A), (S-2A), (D-1A) and (D-2A), the symbols [ka] represents the point of attachment to L; R1 is, independently at each occurrence, -C(O)-A, where A is C1-C6 alkyl, C6-C 18 Aryl, (C6-C 18)aryl(C1-C3)alkyl, 5- to 18-membered heteroaryl, (5- to 18-membered)heteroaryl(C1-C3)alkyl, C3-C8 cycloalkyl, (C3-C8)cycloalkyl(C1-C3)alkyl, 5- to 8-membered heterocycloalkyl, or (5- to 8-membered)heterocycloalkyl(C1-C3)alkyl; one or two carbon atoms within the alkyl portion of A are each independently optionally replaced with a heteroatom selected from N, O, or S. (Examples of alkyl portions of A containing heteroatoms are CH3-CH2-O-CH2-CH2-, CH3-NH-CH2-, CH3-N(CH3)-CH2-, CH3-S-CH2-CH2-, etc.) A is selected from the group consisting of 1 to 3 R 11 groups, and each of the R 11 The group is C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, halogen, C6-C 12 aryl, 5- to 12-membered heteroaryl, cyano, or two groups R 11 together with the atom to which they are attached form a 5- to 7-membered heterocyclyl having 1 to 3 heteroatoms selected from N, O, or S; R 11 are each independently optionally substituted with 1 to 3 substituents selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or cyano.

[0201] In a second aspect of the second example embodiment, the moiety G has the following structural formula: [ka] or a pharmaceutically acceptable salt thereof. The remainder of the values ​​and example values ​​are defined above in relation to the first aspect.

[0202] In some embodiments, the biocompatible polymer can comprise at least one biocompatible polymer selected from the group consisting of polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), poly(ethylene glycol), polyethylene oxide, poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (Pluronic F127), polyoxyethylene-polyoxypropylene block copolymer (Pluronic F68), poloxamer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan. For example, the polymer can comprise the copolymer PLGA-PEG. The remainder of the values ​​and example values ​​are defined above in connection with the first and second embodiments.

[0203] In a third aspect, the biocompatible polymer comprises at least one of polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), polyethylene oxide, polyoxyethylene-polyoxypropylene block copolymer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan. The remainder of the values ​​and example values ​​are defined above in connection with the first and second aspects.

[0204] In a fourth aspect, the biocompatible polymer comprises poly(L-lactic acid), poly(D-lactic acid), poly(D / L-lactic acid), copolymers thereof, or combinations thereof. The remainder of the values ​​and example values ​​are defined above in connection with the first and second aspects.

[0205] In a fifth aspect, the biocompatible polymer comprises polyethylene oxide, a polyoxyethylene-polyoxypropylene block copolymer, a copolymer thereof, or a combination thereof. The remainder of the values ​​and example values ​​are defined above in relation to the first and second aspects.

[0206] In a sixth aspect of the second exemplary embodiment, the polymer P has the following structural formula: [ka] and is represented by the symbol [ka] represents the point of attachment of the polymer to the linker L, and further, y is an integer from 0 to 1000, x is an integer from 0 to 1000, and m is an integer from 0 to 450, with the proviso that x and y are not simultaneously 0. The remainder of the values ​​and example values ​​are defined above in connection with the first to fifth aspects.

[0207] In particular examples of the sixth aspect, y is an integer between 0 and 500, x is an integer between 0 and 500, and m is an integer between 0 and 250. For example, in various aspects, m is between 50 and 200, or m is between 75 and 175, or m is between 90 and 150, or m is between 90 and 140, or m is between 100 and 130. In various aspects, x is between 50 and 200, or x is between 75 and 175, or x is between 90 and 150, or x is between 90 and 140, or x is between 100 and 130. In additional aspects, y is between 5 and 100, or y is between 10 and 75, or y is between 15 and 50. Any combination of values ​​of x, y, and m according to the various aspects is also contemplated. For example, m may be 100-130, x may be 100-130, and y may be 15-50.

[0208] In another example of the sixth aspect, x is an integer from 90 to 140; y is an integer from 10 to 75; and m is an integer from 90 to 140.

[0209] The remainder of the values ​​and example values ​​are defined above in relation to the first to fifth aspects.

[0210] In a seventh embodiment, the particles are acid-terminated PLGA (PLGA-COOH); acid-terminated PLGA-PEG copolymers, where the acid moieties terminate the PEG blocks, acid-terminated PLGA-PEG copolymers (PLGA-PEG-COOH); PLGA-PEG copolymers, where the PEG blocks have the following structural formula: [ka] or a PLGA-PEG copolymer (PLGA-PEG-DBCO), wherein the PEG block is represented by the following structural formula: [ka] wherein the dashed line indicates the point of attachment of the terminal moiety to the PEG block.

[0211] In an example of the seventh embodiment, the particles comprise at least one of: a blend of 75% by weight PLGA-COOH and 25% by weight PLGA-PEG-DBCO; a blend of 75% by weight PLGA-COOH and 25% by weight PLGA-PEG-NHS; a blend of 90% by weight PLGA-PEG-COOH and 10% by weight PLGA-PEG-DBCO; and PLGA-PEG-NHS.

[0212] The remainder of the values ​​and example values ​​are defined above in relation to the first to sixth aspects.

[0213] In an eighth aspect of the second example embodiment, the linker L comprises a moiety that is the product of a click chemistry reaction. The remainder of the values ​​and example values ​​are defined above in relation to the first through seventh aspects.

[0214] In a ninth aspect of the second exemplary embodiment, the linker L comprises a moiety represented by any one of the following structural formulas: [ka] each independently represents a point of attachment to an additional portion of the linker L, P, or G. [ka] In the formula, R * is H or methyl, and R # is independently at each occurrence H or C1-C3 alkyl, and R X is —NH—C(O)O—, and R 200 is —H or C1-C3 alkyl. Examples of additional moieties in the linker include the moiety R L An example of an additional moiety of the linker is -O-(C1-C 12 ) alkylenyl, for example the moiety —O—(CH 2 ) 5 . The remainder of the values ​​and example values ​​are defined above in relation to the first to eighth aspects.

[0215] In a tenth aspect of the second exemplary embodiment, the polymer P is PLGA(10k)-PEG(5k). The remainder of the values ​​and example values ​​are defined above in relation to the first through ninth aspects.

[0216] In an eleventh aspect of the second exemplary embodiment, the weight of G per unit weight of P (ligand density) is 10-75 μg / mg, the remainder of the values ​​and example values ​​being as defined above in relation to the first through tenth aspects.

[0217] In a twelfth aspect of the second exemplary embodiment, the weight of G (also referred to herein as sialic acid-containing ligand) per weight of total solids is from about 1 μg / mg to about 1000 μg / mg.

[0218] In a third example embodiment, the present invention is a method of treating a disorder in a subject in need thereof.

[0219] In a first aspect of the third exemplary embodiment, the method includes administering to a subject an effective amount of a composition comprising a compound or particle described herein, or a pharmaceutically acceptable salt thereof, wherein the disorder is selected from cancer, ophthalmic disease, fibrosis, parasitic inflammation, fungal inflammation, viral inflammation, autoimmune inflammation, neurogenic inflammation, neurodegeneration, skin inflammation, renal inflammation, cardiovascular disease, gastrointestinal inflammation, or rheumatic disease.

[0220] In a fourth example embodiment, the present invention is a method of treating a disorder in a subject in need thereof.

[0221] In a first aspect of the fourth exemplary embodiment, the method includes administering to a subject an effective amount of a composition comprising a compound or particle described herein, or a pharmaceutically acceptable salt thereof, wherein the disorder is selected from breast cancer, non-small cell lung cancer (NSCLC), prostate cancer, colorectal cancer, melanoma, pancreatic cancer, myelofibrosis, diabetic retinopathy, idiopathic pulmonary fibrosis, hepatic fibrosis, sickle cell anemia, and acute respiratory distress syndrome (ARDS).

[0222] In a fifth exemplary embodiment, the invention is a pharmaceutical composition comprising a compound or particle, or a pharmaceutically acceptable salt thereof, as described herein in connection with the first and second exemplary embodiments and various aspects and exemplary aspects thereof, in a pharmaceutically acceptable carrier.

[0223] In a sixth exemplary embodiment, the present invention provides a compound having the following structural formula: GLP A method for making particles comprising a molecule represented by

[0224] Values ​​and example values ​​for the variables G, L and P of the sixth example embodiment are defined herein above in connection with the first and second example embodiments and various aspects thereof.

[0225] In a first aspect of the sixth exemplary embodiment, the method comprises: GR F1 (I) The compound represented by structural formula (II) PRF2 (II) (In the formula, R F1 and R F2 are reactive moieties, respectively) under appropriate conditions to form a compound represented by the moiety R F1 and R F2 with each other to form a covalently linked -L- moiety.

[0226] In a second aspect of the sixth example embodiment, R F1 and R F2 is a pair of click reagents, and the conditions are R F1 and R F2 and the linker L is suitable for a click reaction between R F1 and R F2 and a moiety that is the product of a click chemistry reaction between: and Values ​​and example values ​​for the remainder of the variables are as defined in relation to the first aspect.

[0227] In a third aspect, the biocompatible polymer comprises at least one biocompatible polymer selected from the group consisting of polyglycolic acid, poly(L-lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), polyethylene oxide, polyoxyethylene-polyoxypropylene block copolymer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan. The remainder of the values ​​and example values ​​are defined above in connection with the first and second aspects.

[0228] In a seventh exemplary embodiment, the present invention provides a compound having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof.

[0229] In a first aspect of the seventh example embodiment, R is independently at each occurrence -R L -RF and R L are, independently for each occurrence, -O-, -S-, and -NR 100 -, -O-(C1~C 12 ) Alkylenyl-, -S-(C1-C 12 ) Alkylenyl-, -NR 101 -(C1~C 12 ) Alkylenyl-, -NR 101a -O-(C1~C 12 )Alkylenyl-;-O-(CH2CH2O) m -, -O-(CH2CH2O) k -(CH2CH2)-, -NR 102 -X 100 -(C1~C 12 ) Alkylenyl-, -NR 102a -NR 102b -C(O)-(C1~C 12 ) alkylenyl, and R 100 , R 101 , R 101a , R 102 , R 102a and R 102b are each independently H or C1-C3 alkyl, and X 100 is —O— or —NH—, m and k each independently represent an integer of 1 to 12; R F is expressed independently for each occurrence as H, C1-C3 alkyl, -NH2, -NH-Fmoc, -NH-Boc, -NH-CBz, -NH-Troc, -NH-TFA, mono(C1-C3) alkylamino, di(C1-C3 alkyl)amino; -C(O)-R 103 (R 103 is -H, -OH or (C1-C3) alkyl; The following structural formula [ka] or a click chemistry reagent.

[0230] In one aspect of the seventh example embodiment, -R L -R F is not -OH.

[0231] In various additional aspects, values ​​and example values ​​for the variable R are described above in connection with the first through sixth example embodiments and their various aspects.

[0232] In an eighth exemplary embodiment, the present invention provides a compound having the following structural formula: GLP is a particle containing a molecule represented by

[0233] In a first aspect of the eighth example embodiment, P is a biocompatible polymer, L is a covalent linker; and G is a group having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof, [ka] represents the point of attachment to L.

[0234] In additional aspects, values ​​and example values ​​for the variables L and P are described above in connection with the second example embodiment and its various aspects.

[0235] In a second aspect, the biocompatible polymer comprises at least one of polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), polyethylene oxide, polyoxyethylene-polyoxypropylene block copolymer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan. The remainder of the values ​​and example values ​​are defined above in connection with the first and additional aspects of the eighth embodiment.

[0236] In a second aspect, the biocompatible polymer comprises poly(L-lactic acid), poly(D-lactic acid), poly(D / L-lactic acid), copolymers thereof, or combinations thereof. The remainder of the values ​​and example values ​​are defined above in connection with the first and additional aspects of the eighth embodiment.

[0237] In a third aspect, the biocompatible polymer comprises polyethylene oxide, a polyoxyethylene-polyoxypropylene block copolymer, a copolymer thereof, or a combination thereof. The remainder of the values ​​and example values ​​are defined above in connection with the first and additional aspects of the eighth embodiment.

[0238] In a fourth aspect, the polymer scaffold comprises the block copolymer PLGA-PEG. The remainder of the values ​​and example values ​​are defined above in relation to the first and additional aspects of the eighth embodiment.

[0239] In a fifth embodiment, the polymer P has the following structural formula: [ka] and is represented by the symbol [ka] represents the point of attachment of the polymer to the linker L, and further, y is an integer from 0 to 1000, x is an integer from 0 to 1000, and m is an integer from 0 to 450, with the proviso that x and y are not simultaneously 0. The remainder of the values ​​and example values ​​are defined above in relation to the first to fourth and additional aspects of the eighth embodiment. do.

[0240] In particular examples of the fifth aspect, y is an integer between 0 and 500, x is an integer between 0 and 500, and m is an integer between 0 and 250. For example, in various aspects, m is between 50 and 200, or m is between 75 and 175, or m is between 90 and 150, or m is between 90 and 140, or m is between 100 and 130. In various aspects, x is between 50 and 200, or x is between 75 and 175, or x is between 90 and 150, or x is between 90 and 140, or x is between 100 and 130. In additional aspects, y is between 5 and 100, or y is between 10 and 75, or y is between 15 and 50. In an example of the fifth aspect, x is an integer between 90 and 140; y is an integer between 10 and 75; and m is an integer between 90 and 140. The remainder of the values ​​and example values ​​are defined above in relation to the first through fourth and additional aspects of the eighth embodiment.

[0241] In a sixth embodiment, the particles are acid-terminated PLGA (PLGA-COOH); acid-terminated PLGA-PEG copolymers, where the acid moieties terminate the PEG blocks, acid-terminated PLGA-PEG copolymers (PLGA-PEG-COOH); PLGA-PEG copolymers, where the PEG blocks have the following structural formula: [ka] or a PLGA-PEG copolymer (PLGA-PEG-DBCO), wherein the PEG block is represented by the following structural formula: [ka] wherein the dashed line indicates the point of attachment of the terminal moiety to the PEG block.

[0242] The remainder of the values ​​and example values ​​are defined above in relation to the first to fifth and additional aspects of the eighth embodiment.

[0243] In a seventh aspect, the particles comprise at least one of a blend of 75 wt% PLGA-COOH and 25 wt% PLGA-PEG-DBCO; a blend of 75 wt% PLGA-COOH and 25 wt% PLGA-PEG-NHS; a blend of 90 wt% PLGA-PEG-COOH and 10 wt% PLGA-PEG-DBCO; and PLGA-PEG-NHS. The remainder of the values ​​and example values ​​are defined above in connection with the first through fourth and additional aspects of the eighth embodiment.

[0244] In a ninth exemplary embodiment, the invention is a pharmaceutical composition comprising a compound as described in connection with the seventh exemplary embodiment or any of its aspects, or a particle as described in connection with the eighth exemplary embodiment or any of its aspects, or a pharmaceutically acceptable salt thereof, in a pharmaceutically acceptable carrier.

[0245] In a tenth exemplary embodiment, the present invention is a method of treating a disorder in a subject in need thereof. The method comprises administering to the subject a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof described above in connection with the seventh exemplary embodiment and various aspects thereof, or a composition comprising a particle or a pharmaceutically acceptable salt thereof described above in connection with the eighth exemplary embodiment and various aspects thereof, or a pharmaceutical composition of the ninth exemplary embodiment. In one aspect, the disorder is influenza. The influenza can be human influenza or avian influenza.

[0246] In an eleventh exemplary embodiment, the invention is any of the compounds represented by the structural formulas listed in Table 4, or a pharmaceutically acceptable salt thereof, wherein the variable R is —O—(CH)—NH, —O—(CH)—NH-Fmoc, —O—(CH)—N, or —O—(CH)-biotin.

[0247] In a twelfth exemplary embodiment, the present invention is a compound represented by any of the following structural formulas, a pharmaceutically acceptable salt thereof: [ka] [ka] [ka] [ka] wherein, independently at each occurrence, x is an integer from 90 to 140; y is an integer from 10 to 75; and m is an integer from 90 to 140. [Example]

[0248] Example 1A: Synthesis of S-Compound - Procedure A The following reaction scheme was used for the synthesis of 6-sulfo-sialyl Lewis X derivatives (S series of compounds).

[0249] Scheme 1: Attachment of terminally modified fragments [ka] In Scheme 1, the notation S1XX denotes a compound in which the substituent at the 5-position of the terminal sialic acid is one of the fragments set forth in Table 1 below, and the notation S2XX denotes a compound in which the 9-position of the terminal sialic acid is substituted with one of the fragments set forth in Table 1 below.

[0250] I. Synthesis of precursors 30 and 35 Precursor compounds 30 and 35 in Scheme 1 were synthesized as follows.

[0251] 1. Synthesis of galactose building blocks. Scheme 1-1 [ka] The following conditions were used in the reaction shown in Scheme 1-1: a) BzCl, pyridine; b) AcOH, Ac2O, HBr; c) Ag2CO3, acetone:H2O; d) CNCCl3, DBU.

[0252] More specifically, in a 1 L round bottom flask in an ice bath, galactose (30 g, 166 mmol) was dissolved in 400 mL of pyridine. To the cooled solution, benzoyl chloride (154 mL, 1.33 mol) was added dropwise over 1 hour. After all the benzyl chloride was added, the reaction was removed from the ice bath and mixed at room temperature for 2 hours, which resulted in significant salt formation. The reaction was monitored by TLC (30% EtOAc / Hex, R fThe reaction mixture was monitored by a pH of 0.8 (pH 7.0), and upon completion, the salts were removed by vacuum filtration. Excess pyridine was evaporated under reduced pressure, and the crude material was dissolved in EtOAc (1 L), washed sequentially with 5% H2SO4 (5 × 100 mL), saturated CuSO4 (2 × 100 mL), 1 M HCl (2 × 100 mL), and brine (2 × 100 mL), dried over MgSO4, and the solvent was evaporated under reduced pressure. The resulting oil was transferred to a 2 L round-bottom flask equipped with a condenser and a magnetic stir bar and placed in an oil bath. With vigorous stirring, the round-bottom flask was heated to reflux while MeOH (approximately 1 L total) was added. This process resulted in the formation of the desired perbenzoylated galactose 1, which was collected by vacuum filtration and used in the next step without further purification. 1 was suspended in acetic anhydride (50 mL) and acetic acid (80 mL) in an ice bath with vigorous stirring. To this suspension, hydrogen bromide in acetic acid (204 mL, 833 mmol) was added dropwise over 1 h; the mixture was allowed to reach room temperature. After approximately 2 h, the reaction mixture turned translucent orange. When total conversion was observed by TLC (20% EtOAc / Hex, Rf = 0.50), the reaction was diluted with EtOAc (1 L), extracted with DI water (5 × 200 mL), quenched with sodium bicarbonate (100 mL of DI water, followed by solid sodium bicarbonate), brine (2 × 100 mL), dried over MgSO, and reduced in volume to give 2 as a white solid (104 g, 95% over two steps). To a solution of 2 (104 g, 158 mmol) in acetone (300 mL) and water (14.2 mL) was added freshly prepared AgCO (22 g, 79 mmol). The reaction was kept in the dark at room temperature and monitored by TLC (30% EtOAc / Hex, Rf = 0.45). When all starting material was consumed, the reaction was filtered through Celite, reduced in vacuo, dissolved in EtOAc (500 mL), washed with saturated sodium bicarbonate (3 x 100 mL), brine (2 x 100 mL), dried over MgSO4, and reduced in volume to give the desired product 3. The per-benzoylated galactol 3 was dissolved in DCM (316 mL) containing trichloroacetonitrile (48 mL, 470 mmol). To this solution was added DBU dropwise until the pH of the reaction reached approximately 10. The progress of the reaction was monitored by TLC (15% EtOAc / Hex, R f=0.2) and the reaction volume was reduced when all starting material was consumed. The desired product 4 was purified by flash chromatography (15% EtOAc / Hex) to give the desired α-imidate as a white solid (79.05 g, 68% over two steps). 1 The H data were consistent with those reported in the literature.

[0253] 2. Synthesis of N-trifluoroacetyl-glucosamine building blocks. Scheme 1-2 [ka] The following conditions were used in the reactions shown in Scheme 1-2: a) ethyl trifluoroacetate, sodium methoxide; b) AcO; c) hydrozonium acetate; d) CNCCl, DBU; e) 5-azidopentan-1-ol, TMSOTf; f) sodium methoxide; g) benzaldehyde dimethyl acetal; CSA; h) AcO; i) TFA, triethylsilane.

[0254] More specifically, glucosamine hydrochloride (GlcNH2, 10 g, 46.3 mmol) was added to a 500 mL round-bottom flask containing MeOH (130 mL) and sodium methoxide (51 mL of a 1 M stock solution). The solution was stirred at room temperature until the GlcNAH2 dissolved, at which point ethyl trifluoroacetate (6.2 mL, 51 mmol) was added in one portion. The mixture was stirred at room temperature and monitored by TLC (20% MeOH / DCM, Rf = 0.5). Upon completion, the solvent was reduced in vacuo to provide crude material 5. 5 was dissolved in pyridine (70 mL) in an ice bath. To this solution was added acetic anhydride (22 mL, 231.5 mmol) dropwise over 30 minutes, at which time the reaction was allowed to warm to room temperature. TLC (40% EtOAC / hex, Rf = 0.48) indicated the reaction was complete after 2 hours. The crude reaction mixture was diluted with EtOAc (500 mL), and water-insoluble impurities were extracted with HCl (5 × 100 mL) and brine (3 × 100 mL). The organic layer was dried over MgSO and concentrated under reduced pressure to give the tetra-acetate intermediate 6. Crude intermediate 6 was dissolved in THF (156 mL), and to this solution was added a freshly prepared solution of 2 M hydrazonium acetate (27.8 mL, 55.5 mmol) in methanol. Anomeric acetate deprotection was monitored by TLC (1:1 EtOAc / Hex, Rf = 0.47). When all starting material was consumed, the reaction volume was reduced, and crude 7 was dissolved in EtOAc (500 mL), washed with 1 M HCl (3 × 100 mL), brine (2 × 100 mL), dried over MgSO, and concentrated under reduced pressure. Intermediate 7 was dissolved in DCM (176 mL) and trichloroacetonitrile (9.2 mL, 96.6 mmol) was added to the solution. The reaction was mixed at room temperature and the pH was adjusted to 9 by adding DBU. After 1 h, product formation was observed by TLC (40% EtOAc / Hex, Rf = 0.51). The reaction volume was reduced in vacuo and the product was purified by flash chromatography (20 → 40% EtOAc / Hex) to give the desired imidate 8 as a white foam (15.2 g, 52% over four steps). To a 500 mL round bottom was added 7 (12.7 g, 23.2 mmol), 5-azidopentan-1-ol (3.6 g, 28 mmol), and 4 Å molecular sieves (5 g).The components were dissolved in DCM (232 mL) and stirred at room temperature for 30 minutes, after which the reaction was placed in an ice bath for 30 minutes. To the cooled solution was added TMSOTf (840 μL, 4.64 mmol), and the reaction was monitored by TLC (40% EtOAc / Hex, Rf = 0.4). After conversion of the starting material to the glycosylated product, the reaction was quenched by the addition of triethylamine (970 μL, 6.96 mmol), and the product 9 was purified by flash chromatography (35% EtOAc / Hex) to give a white solid (10 g, 85%). The protected advanced intermediate 9 (10 g, 19.5 mmol) was suspended in MeOH (195 mL), and the pH was adjusted to 8 by the addition of sodium methoxide. The desired triol was observed by TLC (10% MeOH / DCM, Rf = 0.45), and the reaction was purified by Dowex 50WX8 H. +The reaction mixture was neutralized with a resin. The reaction was filtered, and MeOH was removed under reduced pressure to give 10 as a pale yellow wax, which was carried on to the next step without further purification. 10 (7.5 g, 19.4 mmol) was dissolved in ACN (190 mL), and to this solution was added benzaldehyde dimethyl acetal (3.2 mL, 21.3 mmol) and camphorsulfonic acid until the pH reached 3. The reaction mixture was transferred to a rotary evaporator, where it was stirred at 50 °C under reduced pressure. The reaction was monitored by TLC (40% acetone / Hex, Rf = 0.6), and after all starting material was consumed, the pH of the solution was adjusted to 7 with triethylamine. The solvent was removed to give the benzylidene intermediate 11 as a brown solid, which was dissolved in DCM (87 mL). Pyridine (6 mL, 77.6 mmol) and acetic anhydride (3.6 mL, 38.8 mmol) were added to the reaction mixture, which was stirred at room temperature. The progress of the reaction was monitored using TLC (40% acetone / Hex, Rf = 0.65). After all starting material was consumed, the reaction was reduced, resuspended in EtOAc (300 mL), washed with 1 M HCl (3 × 50 mL), brine (2 × 50 mL), dried over MgSO, and concentrated under vacuum. The resulting solid was recrystallized using EtOAc and Hex to give the 3-O-Ac product 12 (6.2 g, 67% over three steps). Selective benzylidene ring opening was achieved by dissolving 12 (2 g, 3.88 mmol) and DCM (33 mL). The reaction was placed in an ice bath, and to the cooled solution were added triethylsilane (3.2 mL, 19.4 mmol) and trifluoroacetic acid (1.48 mL, 19.4 mmol). The reaction was monitored by TLC (40% acetone / Hex, Rf=0.62) and when all 12 was consumed, the reaction was diluted with EtOAc (200 mL), extracted with sodium bicarbonate (3×50 mL), brine (2×50 mL), dried over MgSO4, and the crude reaction mixture was purified by flash chromatography (30% acetone / Hex) to provide 13 (1.52 g, 76%) as a clear oil that turned to a white solid after standing.

[0255] 3. Disaccharide glycosylation Schemes 1-3 [ka] The following conditions were used for the reactions shown in Schemes 1-3: TMSOTf, 4A MS.

[0256] More specifically, acceptor 13 (1.52 g, 3 mmol) and donor 4 (2.8 g, 3.75 mmol) were dissolved in DCM (30 mL), and 4Å MS (2 g) was added to the solution. The solution was stirred at room temperature for 30 min and then placed in an ice bath for 30 min. To the cooled solution, TMSOTf (135 μL, 0.75 mmol) was added in one portion. The reaction was kept on ice with vigorous stirring. After 30 min, TLC (30% EtOAc / Hex, Rf = 0.35) indicated the complete disappearance of 4 and 13, and the desired disaccharide 14 had formed. The reaction was quenched with triethylamine (167 μL, 1.2 mmol) and purified by flash chromatography (30% EtOAc / Hex) to yield pure 14 (1.3 g, 40%) as a white foam.

[0257] 4. 6-O-benzyl deprotection Schemes 1-4 [ka] The following conditions were used for the reactions shown in Schemes 1-4: sodium dithiothionite / sodium bromate.

[0258] More specifically, fully protected disaccharide 14 (1.1 g, 1 mmol) was dissolved in 20 mL of EtOAc and 20 mL of HO with vigorous mixing. To the biphasic solution was added sodium dithiothionite (870 mg, 5 mmol). The reaction was placed in an ice bath, and sodium bromate (150 mg, 1 mmol) was added, resulting in the in situ formation of the bromine radical (the solution turned yellow / orange). The reaction was monitored every 20 min by TLC (40% Actone / Hex, Rf = 0.56). Additional sodium bromate (50 mg, 0.3 mol) was added as needed to convert all 14 back to 15. The reaction was quenched by the addition of saturated sodium thiosulfate and purified by flash chromatography (30% acetone / hex) to give the desired 3-OH product (0.62 g, 62%) as a white solid foam.

[0259] 5.6-O-Sulfation, Global Deprotection, and Amine Acetylation Schemes 1-5 [ka] The following conditions were used in the reactions shown in Schemes 1-5: a) chlorosulfonic acid, TEA; b) NaOH; c) Ac2O.

[0260] More specifically, 15 (620 mg, 0.61 mmol) was dissolved in THF (5 mL) along with triethylamine (1.3 mL, 9.2 mmol) in an ice bath, to which chlorosulfonic acid (405 μL, 6.1 mmol) was added. The reaction was mixed on ice for 1 h, then brought to room temperature and mixed overnight. The reaction was monitored by TLC (10% MeOH / DCM, Rf = 0.58) and desalted by LH-20 size exclusion chromatography (eluted with 1:1 MeOH / DCM). The recovered 3-O-sulfated 16 was suspended in MeOH (8 mL) and HO (4 mL). NaOH (1.1 mL, 5.49 mmol, 5 M stock solution) was added to the reaction mixture, and the solution was stirred vigorously at room temperature, resulting in total deprotection to provide 17, which was observed by TLC (EtOH:NHOH:HO - 7:3:2, Rf = 0.82). The pH of the reaction was adjusted with concentrated AcOH, and the solvent was reduced by rotary evaporation. Crude deprotected 17 was redissolved in MeOH (10 mL), and to this solution were added acetic anhydride (115 μL, 1.22 mmol) and triethylamine (170 μL, 1.22 mmol). Amine acetylation was achieved by mixing at room temperature. 6-Sulfo-N-acetyllactosamine (18) was purified by Bio-Gel P-2 size chromatography using ammonium bicarbonate (100 mM) elution. Product-containing fractions were lyophilized to afford the desired product as a white fluffy solid (203 mg, 55% over three steps).

[0261] 6. Chemical synthesis of 6-sulfo-N-acetylglucosamine Schemes 1-6 [ka] The following conditions were used in the reactions shown in Schemes 1-6: a) AcO, sodium acetate; b) TMSOTf; c) 5-azidopentan-1-ol, TMSOTf; d) sodium methoxide; e) sulfur trioxide-pyridine.

[0262] More specifically, N-acetylglucosamine (100 g, 450 mmol) was added portionwise to a refluxing solution of acetic anhydride (340 mL, 3375 mmol) and sodium acetate (37.2 g, 450 mmol). The mixture was stirred under reflux until all materials dissolved. TLC (100% EtOAc, Rf = 0.47) indicated the reaction was complete. Excess acetic anhydride was removed under reduced pressure, and the crude material was dissolved in EtOAc (1 L), washed with saturated sodium bicarbonate (10 × 150 mL), brine (3 × 100), dried over MgSO4, and reduced in vacuo to provide peracetylated N-acetylglucosamine (19, 129 g, 74%) as a white solid. 19 (5.4 g, 13.9 mmol) was dissolved in DCE (69 mL), and to this solution was added TMSOTf (2.5 mL, 13.87 mmol). The solution was warmed to 60 °C to promote oxazoline formation, which was monitored by TLC (40% acetone / hex, Rf = 0.3). The reaction was quenched with TEA (2.2 mL, 15.18 mmol), the solvent was reduced, and the product was purified by flash chromatography (40% acetone / hex) to give 20 (3.9 g, 85%) as a clear oil. Tri-acetylated oxazoline 20 (4.36 g, 13.24 mmol) was dissolved in DCE (66 mL). To this solution was added 5-azidopentan-1-ol (3.4 g, 26.48 mmol) and 4Å MS (3 g). The combination was mixed at room temperature, after which TMSOTf (2.4 mL, 13.24 mmol) was added, and the reaction was warmed to 60 °C to promote glycosylation. The reaction was monitored by TLC (90% EtOAc / Hex, Rf = 0.55), and when all starting material was converted to product, the reaction was quenched with TEA (1.84 mL, 13.24 mmol) and cooled to room temperature. The solvent was removed under reduced pressure, and the glycosylated N-acetylglucosamine was recovered by flash chromatography (80 → 90% EtOAc / Hex) to provide 21 (5 g, 80%) as a white solid. 21 (14.38, 31.37 mmol) was dissolved in MeOH (156 mL), and the solution was adjusted to pH 9 with sodium methoxide. The reaction was monitored by TLC (20% MeOH / DCM, Rr = 0.76), and upon completion, the pH was adjusted to 9.0 with Dowex 50WX8 H+ The mixture was neutralized with resin. The resin was removed by filtration, and the MeOH was removed under reduced pressure to give triol 22 (9.3 g, 90%) as a white solid. The 6-O-sulfation was added by dissolving 22 (600 mg, 1.8 mmol) in DMF (36 mL) and placing the mixture in an ice bath. To the cooled solution was added sulfur trioxide-pyridine complex (287 mg, 1.8 mmol). The reaction was stirred overnight at 4 °C for 3 days, with daily additions of sulfur trioxide-pyridine (287 mg, 1.8 mmol). The reaction was monitored by TLC (20% MeOH / DCM, Rf = 0.23) and quenched with an aliquot of NaOH (5 M, pH → 8). The solvent was removed, and the reaction was purified by Bio-Gel size-exclusion chromatography. The product-containing fractions were lyophilized to give 23 (514 mg, 69%) as a white powder.

[0263] 2D of compounds 22 and 23 in Schemes 1-6 1 The H-NMR spectra are shown in Figures 1A and 1B.

[0264] 7. Enzymatic introduction of β1,4-Gal to form 6-sulfo-N-acetyllactosamine Schemes 1-7 [ka] β1,4-Gal incorporation was achieved by dissolving 5 mg of 6S-GlcNAc (23.5 mg, 11.7 μmol) and UDP-galactose (10 mg, 17.6 μmol) in Tris buffer (585 μL, pH 7.3, 0.1 M) containing 10 mM MnCl2. To this solution was added B4GALT1 (50 μg, 1% wt / wt) and calf intestinal alkaline phosphatase (CIAP, 5.85 μL, 1 kU stock). The reaction was incubated at 37°C for 3 h. The reaction progress was monitored by LC-ESI-MS equipped with a Waters XBridge BEH amide column, 2.5 μm, 130 Å, 2.1 × 150 mm (flow rate 0.25 mL / min, A = 10 mM ammonium formate, B = ACN, linear gradient 80% → 60% B over 18 min).

[0265] The reaction was monitored by tracking participants and products using hydrophilic interaction liquid chromatography / electrospray ionization mass spectrometry (HILIC-LC / ESI-MS), and the results of such tracking are shown in Figure 2.

[0266] The remaining starting material was converted to product by adding excess UDP-Gal (5 mg, 5.85 μmol) and B4GALT1 (25 μg). The reaction mixture was filtered through a PALL Nanosep® centrifugal spin filter (3k MWCO), and the filtrate was purified by Bio-Gel P2 size-exclusion chromatography. The product-containing fractions were lyophilized to provide the desired disaccharide 24 (4.8 mg, 70%).

[0267] 8.6S-LacNAc azide reduction and Fmoc introduction Schemes 1-8 [ka] The reactions shown in Schemes 1-8 used the following conditions: a) Pd / C, H2, b) Fmoc-OSu.

[0268] More specifically, 24 (100 mg, 0.17 mmol) was dissolved in a 1:1 mixture of HO / t-BuOH (8 mL) containing 5% wt / wt Pd / C. The reaction vessel was evacuated three times and maintained under an atmosphere of H to promote the reduction of the azide to the primary amine. After completion of the azide reduction, the reaction was filtered through a 0.22 μm syringe filter, and the filtrate was lyophilized to provide 25, which was subsequently dissolved in HO (8 mL) containing NaHCO (0.51 mmol). In a separate vessel, FmocOSu (114 mg, 0.34 mmol) was dissolved in ACN (8 mL). To this solution was added dropwise an aqueous solution containing ACN and 25. The mixture was vigorously vortexed to give 26, the formation of which was monitored by C18 TLC (30% ACN / water; Rf = 0.3). The mixture was reduced in vacuo and the product purified by Bio-Gel P2 size exclusion chromatography to give 26 (103 mg, 77% over two steps). 1 H NMR(600MHz,D2O)δ7.23-7.21(m,4H,FmocH1,H1',H4,H4'),6.95(m,4H,FmocH2,H2',H3,H3'),4.40-4.39(d,J=Hz,1H,Ga lH1),4.21-4.16(m,3H,GlcNAcH1,H6),4.06-4.02(m,2H,FmocH8),3.79-3.78(m,1H,GalH4),3.73-3.40(m,11H,Gal,H2, H3,H5,H6,GlcNAcH2,H3,H4,H5,FmocH7, lipid chain H5a),3.11-3.10(m,1H,H5b),2.67-2.65(m,1.5H,H1),2.32(m,0.5H,H1),1. 79(s,3H,GlcNAc,Ac,CH3),1.16-1.12(m,1.5H,H2),1.00-0.98(m,2H,H4),0.87-0.83(m,1.5H,H3),0.69(m,1H,H2,H3). 13C NMR(151MHz,D2O)δ173.90(GlcNAc,-CO-),157.49(Fmoc,-CO-),143.62(Fmoc C6,C6'),140.76(Fmoc C5,C5'),127.60(Fmoc C3,C3'),127.09(Fmoc C2,C2'),124.86(Fmoc C1,C1'),119.75(Fmoc C4,C4'),102.37(Gal,C1),101.03(GlcNAc,C1),77.19(Gal,C3),75.22(GlcNAc,C4) ,72.47(GlcNAc,C5),72.42(Gal,C5),72.15(GlcNAc,C3),70.95(Gal,C2),70.01(C5) ,68.58(Gal,C4),66.11(GlcNAc,C6),65.88(Fmoc,C8),60.94(Gal,C6),54.99(GlcNA c,C2),46.77(Fmoc,C7),40.27(C1),28.50(C4),28.20(C2),22.21(C3,GlcNAc,CH3).

[0269] 9. Enzymatic introduction of α2,3 Neu5Az Schemes 1-9 [ka] Amine-protected 6-sulfo-LacNAc (26, 50 mg, 63 μmol) and ManNAz (27, 33 mg, 126 μmol) were dissolved in Tris buffer (0.1 M, pH 8.5, 6.3 mL) containing MgCl (20 mM), sodium pyruvate (630 μmol), and cytidine-5'-triphosphate (CTP, 252 μmol). 1 (PmNANA, 660 μg), CMP-sialic acid synthetase 2 (NmCSS, 660 μg) and sialyltransferase 1 3-5(PmST1, 500 μg) was added. The reaction was incubated at 37 °C with gentle agitation for 2 h. The reaction was quenched by the addition of EtOH (8 mL) and frozen. The protein precipitate was removed by centrifugation, and the supernatant was reduced by rotary evaporation. The crude reaction mixture was purified by preparative HPLC (Agilent 1200, Eclipse XDB-C18, 21.2 × 250 mm, 7 μm. Solvent A = 50 mM ammonium bicarbonate; B = ACN, λ = 262 μm) using a linear gradient of 10 to 60% B over 30 min at a flow rate of 20 mL / min. Compound-containing fractions were lyophilized to give trisaccharide 28 (59 mg, 83%). 1 H NMR(600MHz,D2O)δ7.57-7.52(m,2H,FmocH4,H4'),7.41-7.35(m,2H,FmocH1,H1'),7.22-7.14(m,4H,FmocH2,H2',H3,H3'),4.46-4 .45(d,1H,GalH1),4.27-4.15(m,5H,FmocH8,GlcNAcH1,H6),4.00-3.99(m,1H,GlcNAcH4),3.93(m,3H,FmocH7,Neu5Az,Az,-CH2),3 .83-3.75(m,4H,GalH4,Neu5AzH5,H6,H9a),3.62-3.46(m,12H,H5a,GalH3,H5,H6,GlcNAcH2,H3,H5,Neu5AzH4,H7,H8,H9b),3.44-3 .41(t,1H,GalH2),3.26-3.14(m,1H,H5b),2.74(m,1.5H,H1),2.64-2.62(dd,1H,Neu5AzH3a),2.29(m,0.5H,H1),1.81(s,3H,GlcNAc Ac,-CH3),1.71-1.67(t,1H,Neu5AzH3b),1.24(m,1.5H,H2),1.08(m,2H,H4),0.94(m,1.5H,H3),0.67(m,1H,H2,H3). 13C NMR(151MHz,D2O)δ174.12(Neu5Az,C1),173.72(GlcNAc,Ac,-CO-),170.98(Neu5Az,Az,-CO-) ,157.94(Fmoc,C9,-CO-),143.67(Fmoc,C6),140.86(Fmoc,C5),127.85(Fmoc,C3),127.29(Fmo c,C2),124.87(Fmoc,C1),119.96(Fmoc,C4),102.07(Gal,C1),101.01(GlcNAc,C1),99.56(Ne u5Az,C2),77.18(Gal,C3),75.27(GlcNAc,C4),74.95(Neu5Az,C7),72.46(GlcNAc,C5),72.43( GlcNAz,C3),72.18(Gal,C5),71.39(Gal,C4),70.14(C5),69.38(Gal,C2),68.15(Neu5Az,C4) ,67.96(Neu5Az,C8),67.38(Neu5Az,C6),66.17(GlcNAz,C6),65.74(Fmoc,C8),62.46(Neu5Az, C9),60.93(Gal,C6),54.96(GlcNAz,C2),51.83(Neu5Az,Az,CH2),51.74(Neu5Az,C5),46.98(F moc,C7),40.13(C1),39.53(Neu5Az,C3),28.26(C4),28.10(C2),22.11(GlcNAc,Ac,CH3 and びC3).

[0270] 10. Introduction of α1,3 フコースの enzyme スキーム1-10

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[0271] 11. アジド return to the original; ライブラリ generates のためのハンドルの modulation. スキーム1-11

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[0272] The synthesis of 6-sulfo-9-amino-sialyl Lewis X was carried out using the same protocol as described above in connection with Schemes 1-8 to 1-11.

[0273] 12. Construction of 6-sulfo-9-amino-sialyl Lewis X Schemes 1-12 [ka] Compounds 33-351 The 1H-NMR spectrum was obtained: Compound 33: 1 1H NMR (600 MHz, D2O) δ 7.48 - 7.43 (m, 2H, FmocH4, H4’), 7.36 - 7.28 (m, 2H, FmocH1, H1’), 7.15 - 7.10 (m, 4H, FmocH2, H2’, H3, H3’), 4.46 - 4.45 (d, 1H, GalH1), 4.24 - 4.19 (m, 5H, FmocH8, GlcNAcH1, H6), 3.98 - 3.96 (m, 2H, GalH4, GlcNAcH4), 3.87 - 3.82 (m, 2H, FmocH7, Neu5AzH6), 3.74 - 3.71 (t, 1H, Neu5AzH5), 3.62 - 3.46 (m, 12H, H5a, GalH3, H5, H6, GlcNAcH2, H3, H5, Neu5AzH4, H7, H8, H9a), 3.43 - 3.38 (m, 2H, GalH2, Neu5AzH9b), 3.21 - 3.09 (m, 1H, H5b), 2.72 (m, 1.5H, H1), 2.63 - 2.60 (dd, 1H, Neu5AzH3a), 2.29 (m, 0.5H, H1), 1.90 (s, Neu5Az, Az, -CH3), 1.80 (s, 3H, GlcNAc Ac, -CH3), 1.69 - 1.65 (t, 1H, Neu5AzH3b), 1.21 (m, 1.5H, H2), 1.06 (m, 2H, H4), 0.91 (m, 1.5H, H3), 0.67 (m, 1H, H2, H3). 1313C NMR (151 MHz, D2O) δ 174.77 (Neu5Ac, Ac, -CO-), 174.07 (Neu5Ac, C1), 173.69 (GlcNAc, Ac, -CO-), 157.84 (Fmoc, C9, -CO-), 143.65 (Fmoc, C6), 140.83 (Fmoc, C5), 127.79 (Fmoc, C3), 127.23 (Fmoc, C2), 124.86 (Fmoc, C1), 119.92 (Fmoc, C4), 102.06 (Gal, C1), 101.03 (GlcNAc, C1), 99.50 (Neu5Ac, C2), 77.12 (Gal, C3), 75.34 (GlcNAc, C4), 74.96 (Neu5Ac, C7), 72.59 (GlcNAc, C5), 72.44 (GlcNAc, C3), 72.14 (Gal, C5), 70.10 (C5), 69.84 (Gal, C4), 69.37 (Gal, C2), 68.58 (Neu5Ac, C4), 68.31 (Neu5Ac, C8), 67.32 (Neu5Ac, C6), 66.05 (GlcNAc, C, 6), 65.79 (Fmoc, C8), 60.93 (Gal, C6), 54.96 (GlcNAc, C2), 52.97 (Neu5Ac, C9), 51.65 (Neu5Ac, C5), 46.90 (Fmoc, C7), 40.15 (C1), 39.64 (Neu5Ac, C3), 28.32 (C4), 28.12 (C2), 22.13 (GlcNAc, Ac, CH3 and C3), 22.01 (Neu5Ac, Ac, CH3). Compound 34: 1H NMR(600MHz,D2O)δ7.75-7.74(d,2H,FmocH4,H4'),7.53-7.52(d,2H,FmocH1,H1'),7 .35-7.33(t,2H,FmocH3,H3'),7.28-7.25(t,2H,FmocH2,H2'),4.95-4.94(d,1H,FucH 1),4.67-4.61(m,1H,FucH5),4.58(m,0.5H,FmocH8),4.50-4.48(d,1H,GalH1),4.37( m,1.5H,FmocH8),4.36(m,1H,GlcNAcH1),4.25-4.18(m,2H,GlcNAcH6),4.12(m,1H,Fm ocH7),3.97-3.95(m,2H,GalH4,GlcNAcH4),3.87-3.80(m,2H,FucH4,Neu5AzH6),3.76-3.52(m,13H,H5a,FucH2,H3,GalH5,H6,GlcNAcH2,H3,H5,Neu5AcH4,H5,H7,H9a),3. 47-3.36(m,5H,H5b,GalH2,GlcNAcH4,Neu5AcH8,H9b),2.80(m,1.5H,H1),2.63-2.60(dd,1H,Neu5AzH3a),2.33(m,0.5H,H1),1.90(s,Neu5Az,Az,-CH3),1.81(s,3H,GlcNAc Ac,-CH3),1.69-1.65(t,1H,Neu5AcH3b),1.32(m,1.5H,H2),1.15(m,2H,H4),1.14-0.99(m,4.5H,H3,FucH6),0.71(m,1H,H2,H3). 1313C NMR (151 MHz, D2O) δ 174.78 (Neu5Ac, Ac, -CO-), 173.99 (Neu5Ac, C1), 173.36 (GlcNAc, Ac, -CO-), 158.16 (Fmoc, C9, -CO-), 143.77 (Fmoc, C6), 140.94 (Fmoc, C5), 127.91 (Fmoc, C3), 127.36 (Fmoc, C2), 124.89 (Fmoc, C1), 120.03 (Fmoc, C4), 101.06 (Gal, C1), 100.77 (GlcNAc, C1), 99.05 (Neu5Ac, C2), 98.48 (Fuc, C1), 75.37 (Gal, C3), 74.70 (GlcNAc, C4), 74.65 (Neu5Ac, C7), 72.83 (GlcNAc, C5), 72.65 (GlcNAc, C3), 72.62 (Fuc, C4), 71.83 (Gal, C5), 70.21 (C5), 69.56 (Gal, C4), 69.29 (Gal, C2), 69.11 (Fuc, C3), 68.48 (Neu5Ac, C4), 68.12 (Neu5Ac, C8), 67.71 (Fuc, C2), 67.04 (Neu5Ac, C6), 66.62 (Fuc, C5), 65.70 (GlcNAc, C6), 65.62 (Fmoc, C8), 61.31 (Gal, C6), 55.60 (GlcNAc, C2), 53.05 (Neu5Ac, C9), 51.65 (Neu5Ac, C5), 47.19 (Fmoc, C-seven), 40.05 (C1), 39.68 (Neu5Ac, C3), 28.18 (C4), 28.06 (C2), 22.12 (GlcNAc, Ac, CH3, C3), 21.99 (Neu5Ac, Ac, CH3), 15.22 (Fuc, C6). Compound 35: 1H NMR(600MHz,D2O)δ7.76-7.75(d,2H,FmocH4,H4’),7.55-7.54(d,2H,FmocH1,H1’),7.36-7.33(t,2H,FmocH3,H3’),7.29-7.26(t,2H,FmocH2,H2’),4.95(d,1H,FucH1),4.67-4.61(m,1.5H,FucH5,FmocH8),4.47-4.46(m,2.5H,GalH1,FmocH8),4.37(m,1H,GlcNAcH1),4.29-4.15(m,3H,GlcNAcH6,FmocH7),3.99-3.93(m,2H,GalH3,Neu5AzH8),3.88-3.85(m,1H,GlcNAcH3),3.76-3.30(m,19H,H5,FucH2,H3,H4,GalH2,H4,H5,H6,GlcNAcH2,H4,H5,Neu5Az,H4,H5,H6,H7,H8,H9a),2.90-2.87(t,1H,Neu5AcH9b),2.80(m,1.5H,H1),2.67-2.64(dd,1H,Neu5AzH3a),2.34(m,0.5H,H1),1.89(s,Neu5Az,Az,-CH3),1.82(s,3H,GlcNAc Ac,-CH3),1.64-1.60(t,1H,Neu5AcH3b),1.32(m,1.5H,H2),1.15-0.98(m,4.5H,H3,FucH6),0.71(m,1H,H2,H3). 13C NMR(151MHz,D2O)δ174.89(Neu5Ac,Ac,-CO-),174.03(Neu5Ac,C1),173.56(GlcNAc,Ac,-CO-),158.19(Fmo c,C9,-CO-),143.80(Fmoc,C6),140.96(Fmoc,C5),127.92(Fmoc,C3),127.37(Fmoc,C2),124.89(Fmoc,C1) ,120.03(Fmoc,C4),101.10(Gal,C1),100.77(GlcNAc,C1),98.99(Neu5Ac,C2),98.52(Fuc,C1),75.47(Gal ,C3),74.80(GlcNAc,C4),74.58(Neu5Ac,C7),72.70(GlcNAc,C5),72.47(GlcNAc,C3),72.40(Fuc,C4),71. 82(Gal,C5),70.23(C5),70.08(Gal,C4),69.28(Gal,C2),69.11(Fuc,C3),68.05(Neu5Ac,C4),67.83(Neu5 Ac,C8),67.68(Fuc,C2),66.73(Neu5Ac,C6),66.61(Fuc,C5),65.78(GlcNAc,C6),65.59(Fmoc,C8),61.34( Gal,C6),55.67(GlcNAc,C2),51.55(Neu5Ac,C5),47.23(Fmoc,C7),42.24(Neu5Ac,C9),40.24(C1),40.04( Neu5Ac,C3),28.18(C4),28.07(C2),22.12(GlcNAc,Ac,CH3,C3),21.96(Neu5Ac,Ac,CH3),15.23(Fuc,C6).

[0274] 13. General Protocol for Fmoc Deprotection Deprotection of S-series compounds was generally carried out according to Schemes 1-13. After deprotection, compounds S1XX and S2XX shown in Scheme 13 are also referred to herein as S1XX-amine and S2XX-amine (see, for example, Example 8, in which S212-amine and S217-amine are conjugated to nanoparticles).

[0275] Scheme 1-13 [ka] For the reactions shown in Schemes 1-13, the following conditions were used: H2O / triethylamine.

[0276] Specifically, compound S1XX or S2XX (0.15 μmol) was dissolved in a 4:1 (v / v) solution of HO / triethylamine (150 μL). The solution was mixed periodically using vortexing over a 3-hour reaction time. Progress was monitored by ESI-MS, and upon conversion of the starting material to the product, the organic material was extracted with EtOAc (500 μL). The aqueous solution was separated and lyophilized to yield the desired deprotected amine target as a white amorphous solid. This material was used for microarray analysis without further purification.

[0277] II. Library of S-Compounds According to Example 1A - Procedure A Tetrasaccharide (30 or 35, 1 mg, 0.8 μmol) was dissolved in DMF (100 μL) containing diisopropylethylamine (DIPEA, 4 μmol). To this solution, acyl chloride (1.6 μmol) was added, and the reaction was mixed using a vortex for 30 min. The reaction was monitored by ESI. When complete, conversion to the product was achieved, and the product was purified by HPLC chromatography under the following conditions: Agilent 1200 HPLC, Eclipse XDB-C8, 4.6 × 250 mm, 5 μm analytical column, solvent = 50 mM ammonium bicarbonate; B = ACN, λ = 262 μm), linear gradient of 10 → 60% B over 30 min, flow rate 1 mL / min. Fractions containing the desired product were lyophilized to yield the desired product as a white fluffy solid. Table 1 lists the 31 R1-acyl chloride fragments used in the synthesis of the S-series compound library.

[0278] Table 1 [ka]

[0279] The following compounds were synthesized and their identities confirmed: 1Confirmed by H-NMR. For the following compounds, the naming convention is such that R has the following formula: [ka]

[0280] The general procedure referred to in the SXX-Fmoc examples below is General Procedure A.

[0281] Compound S101-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1 H NMR(600MHz,D2O)δ8.29(m,1H),7.94-7.87(m,3H),7.78-7.73(m,3H),7.56-7.50(m,4H),7.37-7.35(t ,2H),7.30-7.27(t,2H),4.95-4.95(d,1H),4.67-4.61(m,1.5H),4.48-4.47(m,2.5H),4.39-4.38(m,1H) ),4.22-4.17(m,3H),3.97-3.95(m,1H),3.88-3.36(m,23H),2.81(m,1.5H),2.64-2.61(dd,1H),2.34(m ,0.5H),1.82(s,3H),1.71-1.67(t,1H),1.34(m,1.5H),1.17(m,2H),1.04-1.00(m,4.5H),0.73(m,1H).

[0282] Compound S102-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,dmso)δ8.74-8.72(t,1H),8.30-8.29(dd,1H),8.14(m,1H),8.00-7.99(m,1H),7.96-7.94(dd,1H),7.89-7.86(m ,3H),7.68-7.63(m,3H),7.55-7.52(m,3H),7.41-7.38(t,2H),7.32-7.30(t,2H),7.25-7.24(t,1H),4.82(d,1H),4.44-4.36( m,4H),4.26-4.17(m,4H),4.12-3.96(m,6H),3.90-3.88(m,1H),3.75(m,1H),3.69-3.46(m,12H),3.38(m,1H),3.25-3.19(m, 3H),2.94(m,1H),2.60-2.59(m,1H),1.77(s,3H),1.44-1.38(m,2H),1.37-1.36(m,2H),1.24-1.21(m,2H),0.99-0.97(d,3H).

[0283] Compound S103-Fmoc

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[0284] Compound S104-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1 H NMR(600MHz,dmso)δ8.82-8.80(t,1H),8.07-8.06(m,1H),7.87-7.86(m,3H),7.77-7.76(d,1H),7.68-7.66( m,3H),7.56(s,1H),7.47-7.44(t,1H),7.41-7.38(t,2H),7.33-7.30(m,3H),7.25-7.23(t,1H),4.82-4.81(d ,1H),4.55-4.35(m,3H),4.26-4.10(m,5H),4.07-3.87(m,5H),3.73(m,1H),3.68-3.37(m,13H),3.24-3.18( m,3H),2.93(m,1H),2.70-2.60(m,1H),1.76(s,3H),1.44-1.35(m,4H),1.24-1.21(m,2H),0.98-0.97(d,3H).

[0285] Compound S105-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,dmso)δ9.13-9.11(t,1H),8.23-8.22(d,1H),8.16-8.15(d,1H),8.10-8.08(m,1H),7.89-7.86(m, 3H),7.68-7.67(d,2H),7.64-7.56(m,2H),7.40-7.38(t,2H),7.32-7.30(t,2H),7.25-7.23(t,1H),4.82-4.81 (d,1H),4.58-4.35(m,3H),4.26-4.03(m,10H),3.94-3.90(m,2H),3.73(m,1H),3.68-3.37(m,13H),3.25-3.18 (m,3H),2.93(m,3H),2.71-2.60(m,1H),1.76(s,3H),1.44-1.34(m,4H),1.25-1.21(m,2H),0.98-0.97(d,3H).

[0286] Compound S106-Fmoc

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[0287] Compound S107-Fmoc

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[0288] Compound S108-Fmoc

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[0289] Compound S109-Fmoc

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[0290] Compound S110-Fmoc

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[0291] Compound S111-Fmoc

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[0292] Compound S112-Fmoc

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[0293] Compound S113-Fmoc

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[0294] Compound S114-Fmoc

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[0295] Compound S115-Fmoc (0.15μmol) is prepared by using the usual formula.

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[0296] Compound S116-Fmoc

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[0297] Compound S117-Fmoc

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[0298] Compound S118-Fmoc

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[0299] Compound S119-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0300] Compound S120-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.78(d,J=7.6Hz,2H),7.56(d,J=7.6Hz,2H),7.36(t,J=7.5Hz,2H),7.29(t,J=7.5Hz,2H) ,4.95(d,J=3.4Hz,1H),4.46(d,J=7.8Hz,1H),4.38(d,J=7.7Hz,1H),4.25-4.16(m,3H),3.95(d,J=10.0Hz,1H ),3.86(t,J=8.8Hz,1H),3.82-3.35(m,26H),2.81(s,2H),2.61(dd,J=12.4,4.6Hz,1H),2.05(s,2H),1.81(s, 1H),1.66(t,J=12.1Hz,1H),1.33(s,1H),1.21-1.18(m,2H),1.02(d,J=6.6Hz,5H),0.85(s,8H),0.72(s,1H).

[0301] Compound S121-Fmoc

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[0302] Compound S122-Fmoc

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[0303] Compound S123-Fmoc

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[0304] Compound S124-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.77(d,J=7.6Hz,2H),7.68(d,J=9.0Hz,2H),7.56(d,J=7.5Hz,2H),7.36(t,J=7.5Hz,2H),7. 28(t,J=7.5Hz,2H),6.94(d,J=9.0Hz,2H),4.94(d,J=4.0Hz,1H),4.46(d,J=8.1Hz,1H),4.38(d,J=7.4Hz,1H),4. 26-4.13(m,3H),4.01-3.92(m,3H),3.85(t,J=8.8Hz,1H),3.83-3.30(m,25H),2.81(s,2H),2.61(dd,J=12.6,4.7 Hz,1H),1.81(s,3H),1.67(t,J=12.2Hz,1H),1.33(s,1H),1.23-1.10(m,6H),1.02(d,J=6.6Hz,5H),0.72(s,1H).

[0305] Compound S125-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.77(d,J=7.6Hz,2H),7.56(d,J=7.5Hz,2H),7.52(s,1H),7.47(d,J=8.0Hz,1H),7.39-7.32( m,3H),7.31-7.25(m,3H),4.94(d,J=4.0Hz,1H),4.46(d,J=7.9Hz,1H),4.38(d,J=7.7Hz,1H),4.26-4.16(m,3H) ,4.03-3.91(m,3H),3.86(t,J=8.9Hz,1H),3.83-3.22(m,22H),2.81(s,2H),2.61(dd,J=12.5,4.7Hz,1H),2.26( s,3H),1.81(s,1H),1.67(t,J=12.2Hz,1H),1.33(s,1H),1.21-1.11(m,4H),1.02(d,J=6.6Hz,5H),0.72(s,1H).

[0306] Compound S126-Fmoc

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[0307] Compound S127-Fmoc

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[0308] Compound S128-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0309] Compound S129-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.78(d,J=7.6Hz,2H),7.57(dd,J=17.6,7.3Hz,3H),7.36(t,J=7.5Hz,2H),7.28(t,J=7.5Hz,2H ),7.04(t,J=9.1Hz,1H),4.94(d,J=4.0Hz,1H),4.46(d,J=8.1Hz,1H),4.38(d,J=5.7Hz,2H),4.25-4.14(m,7H),4. 01-3.92(m,6H),3.86(t,J=8.7Hz,1H),3.83-3.32(m,23H),2.81(t,J=5.4Hz,3H),2.61(dd,J=12.1,4.4Hz,1H),2. 17(s,2H),1.81(s,2H),1.67(t,J=12.2Hz,1H),1.33(s,1H),1.20-1.13(m,6H),1.02(d,J=6.6Hz,5H),0.72(s,1H).

[0310] Compound S130-Fmoc

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[0311] Compound S131-Fmoc

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[0312] The following compounds were synthesized according to the general procedure:

[0313] Compound S201-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0314] Compound S202-Fmoc (0.135 μmol) was prepared using the general procedure. [ka]

[0315] Compound S203-Fmoc (0.135 μmol) was prepared using the general procedure. [ka]

[0316] Compound S204-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0317] Compound S205-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0318] Compound S206-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0319] Compound S207-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0320] Compound S208-Fmoc (0.075 μmol) was prepared using the general procedure. [ka]

[0321] Compound S209-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0322] Compound S210-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0323] Compound S211-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0324] Compound S212-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0325] Compound S213-Fmoc [ka] (0.075 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.74(d,J=7.5Hz,2H),7.53(d,J=8.4Hz,5H),7.34(t,J=7.5Hz,2H),7.30-7. 24(m,2H),7.13(d,J=6.4Hz,3H),4.93(s,1H),4.43(t,J=7.2Hz,4H),4.34(s,1H),4.24-4.04(m, 2H),3.95-3.36(m,26H),2.90-2.75(m,4H),2.62(d,J=10.4Hz,1H),2.18(s,3H),1.89-1.78(m,1 0H),1.63(t,J=12.3Hz,1H),1.35(s,1H),1.20-1.19(m,2H),1.02(d,J=6.6Hz,5H),0.73(s,1H).

[0326] The following compounds were synthesized according to the general procedure:

[0327] Compound S214-Fmoc (0.075 μmol) was prepared using the general procedure. [ka]

[0328] Compound S215-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0329] Compound S216-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0330] Compound S217-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0331] Compound-Fmoc (0.15 μmol) was prepared using the general procedure. S218 [ka]

[0332] Compound S219-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0333] Compound S220-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.78(d,J=7.6Hz,2H),7.56(d,J=7.5Hz,2H),7.36(t,J=7.5Hz,2H),7.28(t,J=7.5Hz,2H),4.94(d,J=3.9 Hz,1H),4.47(d,J=8.1Hz,1H),4.39(d,J=5.6Hz,1H),4.27(d,J=10.5Hz,1H),4.19(t,J=13.0Hz,2H),3.92(dd,J=9.6,2.9Hz ,2H),3.90-3.82(m,2H),3.81-3.20(m,22H),2.81(s,2H),2.61(dd,J=12.5,4.8Hz,1H),2.12-1.95(m,2H),1.90-1.85(m,3H ),1.82(s,2H),1.62(t,J=12.2Hz,1H),1.34(s,1H),1.19(d,J=6.6Hz,2H),1.03(d,J=6.5Hz,4H),0.84(s,5H),0.73(s,2H).

[0334] Compound S221-Fmoc

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[0335] Compound-Fmoc

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[0336] Compound S223-Fmoc (0.15 μmol) was prepared using the general procedure. [ka] 1H NMR(600MHz,D2O)δ7.93-7.42(m,8H),7.37-7.29(m,2H),7.26(t,J=7.5Hz,2H),4.93(d ,J=4.0Hz,1H),4.48-4.40(m,1H),4.37(d,J=6.0Hz,1H),4.21-4.06(m,2H),4.00-3.32( m,26H),2.91-2.74(m,2H),2.63(dd,J=12.5,4.6Hz,1H),1.87(s,3H),1.82(s,3H),1.6 3(t,J=12.2Hz,1H),1.34(s,1H),1.20-1.19(m,2H),1.02(d,J=6.6Hz,5H),0.73(s,1H).

[0337] Compound S224-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.71(d,J=7.7Hz,2H),7.67-7.47(m,4H),7.32(q,J=8.6,8.1Hz,2H),7.26(q,J=9 .2,7.5Hz,2H),6.97-6.75(m,2H),4.93(d,J=3.9Hz,1H),4.50-4.38(m,1H),4.37-4.28(m,1H),4.18 -4.04(m,2H),4.03-3.21(m,28H),2.90-2.77(m,2H),2.62(dd,J=12.4,4.5Hz,1H),1.86(s,3H),1.8 2(s,3H),1.63(t,J=12.2Hz,1H),1.35(s,1H),1.20-1.17(m,2H),1.02(d,J=6.6Hz,5H),0.74(s,1H).

[0338] Compound S225-Fmoc

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[0339] Compound S226-Fmoc

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[0340] Compound S227-Fmoc

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[0341] Compound S228-Fmoc (0.15μmol) is used, and it is prepared by using a general hand-operated method.

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[0342] Compound S229-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ7.75(dd,J=32.6,7.7Hz,1H),7.62-7.39(m,4H),7.30(dq,J=43.3,9.1,7.4Hz ,4H),7.08-6.87(m,1H),4.93(d,J=4.0Hz,1H),4.51-4.29(m,2H),4.26-4.06(m,2H),4.02-3.24( m,26H),2.91-2.76(m,1H),2.62(dd,J=12.4,4.5Hz,1H),2.08(s,3H),1.84(d,J=28.4Hz,6H),1.6 3(t,J=12.2Hz,1H),1.38-1.32(m,1H),1.20(d,J=6.5Hz,2H),1.02(d,J=6.6Hz,5H),0.73(s,1H).

[0343] Compound S230-Fmoc (0.15 μmol) was prepared using the general procedure. [ka]

[0344] Compound S231-Fmoc [ka] (0.15 μmol) was prepared using the general procedure. 1H NMR(600MHz,D2O)δ8.00-7.79(m,2H),7.69(dd,J=31.0,7.9Hz,3H),7.59-7.39(m,3H),7.37-7 .20(m,4H),4.93(d,J=4.0Hz,1H),4.47-4.40(m,1H),4.39-4.33(m,1H),4.22-4.06(m,2H),4. 03-3.15(m,26H),2.90-2.75(m,1H),2.63(dd,J=12.4,4.6Hz,1H),1.88(s,3H),1.82(s,3H),1 .63(t,J=12.2Hz,1H),1.34(s,1H),1.19(d,J=6.6Hz,2H),1.02(d,J=6.7Hz,5H),0.74(s,1H).

[0345] Example 1B - Synthesis of S-Compound - Procedure B I.Synthesis The following reaction scheme was used for the synthesis of 6-sulfo-sialyl Lewis X derivatives (S series of compounds).

[0346] Scheme 10: Attachment of terminally modified fragments [ka] In Scheme 10, the designation S1XX denotes a compound in which the substituent at the 5-position of the terminal sialic acid is one of the fragments set forth in Table 1 herein, and the designation S2XX denotes a compound in which the 9-position of the terminal sialic acid is substituted with one of the fragments set forth in Table 1. All compounds made according to Scheme 10 are [ka] and is referred to herein as S1XX-azide, etc.

[0347] I. Synthesis of Precursors 300 and 350 Precursor compounds 300 and 350 of Scheme 10 were synthesized as follows.

[0348] 1. Synthesis of Galactose Building Blocks Scheme 10-1 [ka] The following conditions were used in the reaction shown in Scheme 10-1: a) BzCl, pyridine; b) AcOH, Ac2O, HBr; c) Ag2CO3, acetone:HO; d) CNCCl3, DBU. Note that Scheme 10-1 shows the same transformation as Scheme 1-1 in Procedure A. The compound numbering in Scheme 10-1 is different from Scheme 1-1, but the compounds are the same.

[0349] More specifically, galactose (30 g, 166 mmol) was dissolved in 400 mL of pyridine in a 1 L round-bottom flask placed in an ice bath. To this cooled solution, benzoyl chloride (154 mL, 1.33 mol) was added dropwise over 1 h. After all the benzoyl chloride had been added, the reaction was removed from the ice bath and allowed to mix at room temperature for 2 h, which resulted in significant salt formation. The reaction was monitored by TLC (30% EtOAc / Hex, Rf = 0.8), and upon completion, the salt was removed by vacuum filtration. Excess pyridine was evaporated under reduced pressure, and the crude material was dissolved in EtOAc (1 L) and washed sequentially with 5% H2SO4 (5 x 100 mL), saturated CuSO4 (2 x 100 mL), 1 M HCl (2 x 100 mL), and brine (2 x 100 mL), dried over MgSO4, and the solvent was evaporated under reduced pressure. The resulting oil was transferred to a 2 L round-bottom flask equipped with a condenser and a magnetic stir bar and placed in an oil bath. With vigorous stirring, the round-bottom flask was heated to reflux while MeOH (approximately 1 L total) was added. This process resulted in the formation of the desired perbenzoylated galactose 10, which was collected by vacuum filtration and used in the next step without further purification. In an ice bath, 10 was suspended in acetic anhydride (50 mL) and acetic acid (80 mL) with vigorous stirring. To this suspension, hydrogen bromide in acetic acid (204 mL, 833 mmol) was added dropwise over 1 h; the mixture was allowed to reach room temperature. After approximately 2 h, the reaction mixture turned translucent orange. When complete conversion was observed by TLC (20% EtOAc / Hex, Rf = 0.50), the reaction was diluted with EtOAc (1 L), extracted with DI water (5 × 200 mL), quenched with sodium bicarbonate (100 mL of DI water, followed by solid sodium bicarbonate), brine (2 × 100 mL), dried over MgSO, and reduced in volume to give 20 as a white solid (104 g, 95% over two steps). To a solution of 20 (104 g, 158 mmol) in acetone (300 mL) and water (14.2 mL) was added freshly prepared AgCO (22 g, 79 mmol). The reaction was kept in the dark at room temperature and monitored by TLC (30% EtOAc / Hex, Rf = 0.45).When all starting material had been consumed, the reaction was filtered through Celite, reduced in vacuo, dissolved in EtOAc (500 mL), washed with saturated sodium bicarbonate (3 × 100 mL), brine (2 × 100 mL), dried over MgSO, and reduced in volume to give the desired product 3. Perbenzoylated galactol 30 was dissolved in DCM (316 mL) containing trichloroacetonitrile (48 mL, 470 mmol). DBU was added dropwise to this solution until the pH of the reaction reached approximately 10. The reaction progress was monitored by TLC (15% EtOAc / Hex, Rf = 0.2), and the reaction volume was reduced when all starting material had been consumed. The desired product 40 was purified by flash chromatography (15% EtOAc / Hex) to give the desired α-imidate as a white solid (79.05 g, 68% over two steps). 1 The H data were consistent with those reported in the literature.

[0350] 2. Synthesis of N-phthalimido-glucosamine building blocks Scheme 10-2 [ka] The following conditions were used in the reaction shown in Scheme 10-2: a) phthalic anhydride, sodium methoxide; b) acetic anhydride, pyridine; c) ethylenediamine / acetic acid; d) trichloroacetonitrile, DBU; e) 5-amino-Cbz-pentan-1-ol, TMSOTf; f) sodium methoxide; g) TBDPS-Cl, imidazole.

[0351] Glucosamine hydrochloride (20 g, 93 mmol) was dissolved in MeOH (100 mL) containing sodium methoxide (1 M, 110 mL) and mixed at room temperature for 30 minutes. The reaction mixture was filtered to remove NaCl, and the filtrate was added to a 500 mL round-bottom flask. To this solution was added phthalic anhydride (15.1 g, 102 mmol), and the mixture was mixed vigorously under reflux for 2 hours. During this time, the product formed as a white, insoluble material. After 2 hours, the reaction was cooled in an ice bath, and the insoluble material (50) was filtered, washed with cold MeOH, and dried under reduced pressure. The solid material (50) was suspended in pyridine (130 mL) at 4 °C, and to this solution was added acetic anhydride (52 mL, 560 mmol) dropwise, followed by catalytic DMAP. The mixture was kept on ice until all the acetic anhydride had been added, after which the reaction was allowed to warm to room temperature and mix overnight. The reaction progress was monitored by TLC (1:1 EtOAc / Hex), and upon completion, excess pyridine and acetic anhydride were removed under reduced pressure. The resulting crude material was dissolved in EtOAc (1 L) and washed thoroughly with 1 M HCl (10 × 150 mL), sodium bicarbonate (5 × 100 mL), brine (2 × 100 mL), and dried over MgSO. The solvent was removed under reduced pressure to give a white solid 60 (35 g), which was carried on to the next step without further purification. Ethylenediamine (5.88 mL, 88 mmol) and acetic acid (5 mL, 88 mmol) were dissolved in THF (366 mL) at room temperature and mixed for 30 min. To this solution was added 60 (35 g), and the solution was left stirring overnight. The reaction was monitored by TLC (1:1 EtOAc / Hex) and upon completion, THF was removed under reduced pressure and the crude material was redissolved in EtOAc (500 mL), washed with 1 M HCl (5 × 75 mL), saturated sodium bicarbonate (3 × 50 mL), brine (2 × 50 mL), dried over MgSO4, and reduced in vacuo to provide 70 as a white solid. 70 (27.1 g) was dissolved in DCM (311 mL) containing trichloroacetonitrile (7.5 mL, 75 mmol). To this mixture was added DBU dropwise until the reaction reached pH → 10.The progress of the reaction was monitored by TLC (1:1 EtOAc / Hex) and upon completion, the solvent was removed under reduced pressure and the product purified by flash chromatography (1:1 EtOAc / Hex containing 0.1% TEA) to give 80 as a clear oil (25.2 g, 46% yield over four steps). 1 H NMR and mass spectrometry data were consistent with the details reported in the literature. 80 (12.6 g, 21.7 mmol) and 5-amino-Cbz-pentan-1-ol (5.15 g, 21.7 mmol) were dissolved in DCM (217 mL) containing flame-dried 4 Å molecular sieves (5 g, Sigma-Aldrich 208590). The solution was mixed at room temperature for 30 min and then placed in an ice bath at 4 °C for 30 min. To the ice-cold solution was added TMSOTf (786 μL, 4.35 mmol), and progress was monitored by TLC (40% EtOAc / Hex). When all the starting imidate was consumed, the reaction was quenched by the addition of triethylamine (1.5 mL, 10.85 mmol). The reaction was filtered through a pad of Celite, reduced in vacuo, and purified by flash chromatography (30→40% EtOAc / Hex) to give 90 (8 g, 57%) as a viscous oil. 1H NMR and mass spectrometry data were consistent with details reported in the literature. 90 (8 g, 12.2 mmol) was dissolved in MeOH (120 mL), and sodium methoxide in methanol (1 M stock solution) was added dropwise to this solution until the reaction pH stabilized at approximately 9. Care was taken not to allow the pH to become basic above 9 due to the risk of phenylimide ring-opening. The reaction was monitored by TLC (10% MeOH / DCM) and confirmed by MALDI-MS. When all acetate had been removed, the reaction pH was neutralized using an Amberchrom 50WX8 hydrogen form, 200-400 mesh (Sigma #217514). The resin was isolated by vacuum filtration, and the reaction solvent was removed under reduced pressure to yield the desired triol 100 as a sticky white foam. To 100 was added DMF (120 mL) and imidazole (1.66 g, 24.4 mmol). The solution was placed in an ice bath, and TBDPS-Cl (4.7 mL, 18.3 mmol) was added dropwise to the mixture over 1 h. The reaction was allowed to warm to room temperature overnight, and the next day the reaction was monitored by TLC (100% EtOAc). DMF was removed by rotary evaporation, and the crude product was redissolved in EtOAc (500 mL). The organic layer was washed three times with 1 M HCl (3 × 50 mL), brine (2 × 50 mL), dried over MgSO4, reduced, and purified by flash chromatography (40% EtOAc / Hex) to provide 110 (6.53 g, 70% over two steps) as a white foam. 1 1 H NMR and mass spectrometry data were consistent with details reported in the literature.

[0352] 3. Disaccharide glycosylation Scheme 10-3 [ka] The following conditions were used in the reaction shown in Scheme 10-3: TMSOTf, 4A MS.

[0353] More specifically, acceptor 110 (6.53 g, 8.5 mmol) and donor 40 (6.3 g, 8.5 mmol) were dissolved in DCM (85 mL) containing 4 Å MS and mixed at room temperature for 30 min. The reaction mixture was cooled to −40 °C in a dry ice bath, and TMSOTf (308 μL, 1.7 mmol) was added to the mixture. The reaction was slowly warmed to −20 °C for 1 h. During this time, the reaction was monitored by TLC (40% EtOAc / Hex) and, upon completion, quenched with TEA (474 ​​μL, 3.4 mmol). Note: This reaction is a regioselective glycosylation. Overglycosylation at the 3-position is possible if the reaction temperature is too high or if an excess of donor 40 is used. The reaction was filtered through Celite and purified by flash chromatography (40% EtOAc / Hex) to give the product 120 as a white foam (5.1 g, 44% yield). 1 H(600MHz,CDCl3),δ(ppm):GlcNPhth:5.13(H1),4.18(dd,J=10.8,8.6Hz,H2),4.60(H3),4.09(H4),3.46(H5),3.79(H6a,H6b);Gal:5.11(H1),5.85( dd,J=10.5,8.1Hz,H2),5.57(dd,J=10.5,3.5Hz,H3),5.95(d,J=3.3Hz,H4 ),4.29(H5),4.69(ddd,J=11.7,4.3,H6a),4.37(ddd,J=11.6,8.3Hz,H6b). from HSQC (75MHz, CDCl3) 13 C, δ(ppm):GlcNPhth:97.8(C1),56.1(C2),69.8(C3)80.2(C4),75.3(C5),61. 7(C6);Gal:101.5(C1),69.6(C2),71.6(C3),67.9(C4),72.5(C5),62.6(C6).

[0354] MALDI-TOF MS C 77 H 76 N2O 18 SiNa(M+Na) + Calculated m / z for = 1367.4755, Found = 1367.6501

[0355] 4. Debenzoylation, phthalimide deprotection, and global acetylation Scheme 10-4 [ka] The reaction shown in Scheme 10-4 used the following conditions: a) sodium methoxide in methanol; b) hydrazine hydrate; c) acetic anhydride and pyridine.

[0356] More specifically, disaccharide 120 (4.7 g, 3.5 mmol) was dissolved in MeOH (175 mL), and aliquots of sodium methoxide (1 M stock solution) were added to the reaction until the reaction pH reached approximately 9. Care was taken to ensure the pH did not exceed 9, as increased basicity could result in undesired imide ring opening. The reaction progress was monitored by MALDI-MS. When all benzoyl groups were removed, the reaction pH was neutralized with Amberchrom 50WX8 hydrogen form, the resin was removed by vacuum filtration, and the MeOH was reduced under reduced pressure to give 130, which was carried on to the next step without further purification. 130 was dissolved in EtOH (175 mL), and hydrazine hydrate (50-60%, 1.6 mL, 17.5 mmol) was added to the solution. The reaction was stirred at reflux until phenylimide deprotection was achieved (monitored by MALDI-MS). After successful phalimide deprotection, the reaction was cooled, the insoluble material was filtered through Celite, and the filtrate was reduced to give crude 140, which was carried forward without further purification. 140 was dissolved in pyridine (65 mL) and catalytic DMAP was added to the reaction. The reaction was placed in an ice bath and acetic anhydride (6.6 mL, 70 mmol) was added dropwise. After all the anhydride had been added, the reaction was allowed to warm to room temperature. The reaction was monitored by TLC (40% EtOAc / Hex). After conversion to the product, the reaction was quenched with MeOH (20 mL), the solvent was reduced in vacuo, and the crude material was redissolved in EtOAc (500 mL), washed with 1 M HCl (10 × 50 mL), saturated copper sulfate (2 × 50 mL), saturated sodium bicarbonate (5 × 50 mL), brine (2 × 50 mL), dried over MgSO, reduced, and purified by flash chromatography (40% EtOAc / Hex) to give 150 (2.03 g, 55% over 3 steps) as a white foam. 1H(600MHz,CDCl3),δ(ppm):GlcNAc:4.35(d,J=7.8Hz,H1),4.07(H2),5.01(H3),4.11(H4),3.31(H5),3.94(H6a),3.89(H6b); Gal:4.76(d,J=8.0Hz,H1),5.06(H2),4.92(dd,J=3.5Hz,H3),5.31(dd,J=3.2Hz,H4),3.76(dd,J=6.6Hz,H5),4.11(H6a,H6b) from HSQC (75MHz, CDCl3) 13 C:δ(ppm):GlcNAc:101.1(C1),53.6(C2),72.3(C3),73.9(C4),75.4(C5),61. 3(C6);Gal:100.5(C1),69.3(C2),71.0(C3),67.2(C4),70.7(C5),61.3(C6).

[0357] MALDI-TOF MS C 53 H 70 N2NaO 18 SiNa(M+Na) + m / z calculated for = 1073.4285, observed = 1073.5575

[0358] 5.6-O-TBDPS deprotection. Scheme 10-5 [ka] The following conditions were used in the reaction shown in Scheme 10-5: a) HF-pyridine

[0359] More specifically, 150 (2.03 g, 1.93 mmol) was dissolved in pyridine (15 mL) and placed in an ice bath. To this solution was added HF-pyridine (70%, 2.3 mL, 19.3 mmol). When all of the HF-pyridine had been added, the reaction was kept on ice for 30 minutes and then warmed to room temperature. The reaction was monitored by TLC (40% acetone / hex), and upon completion, the contents were transferred to EtOAc (300 mL), extracted with 1 M HCl, brine, and dried over MgSO. The crude reaction was purified by flash chromatography using 1:1 acetone / hex to give 160 (1.12 g, 72%) as a white foam. 1 H(600MHz,CDCl3),δ(ppm):GlcNAc:4.40(d,J=8.2Hz,H1),3.96(H2),5.06(H3),3.91(H4),3.35(H5),3.86(H6a),3.72(H6b );Gal:4.62(d,J=7.9Hz,H1),5.11(H2),4.99(H3),5.34(dd,J=2.5Hz,H4),3.90(H5),4.09(ddd,J=11.1,5.9Hz,H6a and H6b). From HSQC 13 C(75MHz,CDCl3),δ(ppm):GlcNAc:101.3(C1),54.1(C2),73.0(C3),74.9(C4),75.0( C5),60.7(C6);Gal:100.9(C1),69.8(C2),70.9(C3),68.0(C4),70.4(C5),61.2(C6).

[0360] MALDI-TOF MS C 37 H 52 N2O 18 Na(M+Na) + m / z calculated for = 835.3113, found = 835.4063

[0361] Introduction of 6.6-O sulfate Scheme 10-6 [ka] The following conditions were used in the reaction shown in Scheme 10-6: a) sulfur trioxide pyridine complex in DMF

[0362] More specifically, 160 (1.34 g, 1.65 mmol) was dissolved in 33 mL of DMF at room temperature. To this mixture was added sulfur trioxide pyridine complex (787 mg, 4.9 mmol), and the reaction was stirred overnight. Product formation was monitored by TLC (10% MeOH / DCM). When all starting material was consumed, the reaction was quenched by adding 3 mL of a 1:1 mixture of MeOH / TEA. The solvent was removed under reduced pressure, and the product was desalted using size exclusion chromatography (2.5 × 100 cm, elution with 1:1 MeOH / DCM, 4 min / fraction, 4–5 mL / fraction) on an LH-20 column. The product was found to elute in fractions 66–76 and, after drying, yielded a white foam. Product 170 was carried on to the next step without further purification.

[0363] 7. Global deacetylation Scheme 10-7 [ka] The following conditions were used in the reaction shown in Scheme 10-7: a) sodium methoxide in methanol.

[0364] More specifically, 170 (1.25 g, 12.8 mmol) was dissolved in MeOH (50 mL) and the pH was adjusted to approximately 9 with sodium methoxide in methanol (1 M stock). The progress of the reaction was monitored by TLC (7:3:2 EtOH:NH4OH:HO) and ESI-MS. When all acetate groups had been removed, the reaction pH was adjusted using Amberchrom H as previously described. +The product was neutralized with resin, which was then removed by vacuum filtration. The filtrate was dried under reduced pressure, and the corresponding solid was dissolved in a minimal volume of 100 mM ammonium bicarbonate. This material was applied to a Bio-Rad P2 size-exclusion column (60 x 5 cm) and eluted with 100 mM ammonium bicarbonate, with each fraction showing a 15-minute elution (approximately 40 mL / fraction). Fractions were stained with 5% sulfuric acid in ethanol and confirmed by ESI. The product-containing fractions were collected, concentrated under reduced pressure, and placed on a lyophilizer to give 180 (726 mg, 80% yield) as a white, fluffy solid. 1 H(600MHz,CDCl3),δ(ppm):GlcNAc:4.38(H1),3.58(H2),3.63(H5),4.26(H6a),4.17 (H6b), 1.85 (NHAc); Gal: 4.38 (H1), 3.38 (H2), 3.55 (H3), 3.79 (H4), 3.62 (H6a and H6b). From HSQC 13 C(75MHz,CDCl3),δ(ppm):GlcNAc:102.1(C1),55.2(C2),72.6(C5),66.0(C6),22.1(NHAc);Gal:102.1(C1),71.2(C2),72.3(C3),68.8(C4),61.0(C6)

[0365] ESI-TOF MS C 27 H 41 N2O 16 S(MH) - m / z calculated for = 681.2182, found = 681.0994

[0366] 8.Cbz deprotection Scheme 10-8 [ka] The following conditions were used in the reaction shown in Scheme 10-8: a) Pd / C, H2, 1:1 t-BuOH / H2O.

[0367] More specifically, 180 (100 mg, 0.14 mmol) was dissolved in a 1:1 mixture of t-BuOH / HO (1.5 mL), and to this solution was added a catalyst, approximately 5% (wt / wt) Pd / C. The reaction was stirred overnight under a hydrogen atmosphere. The reaction progress was monitored by TLC (7:3:2 EtOH:NHOH:HO), and upon completion, the reaction was filtered through a 0.2 μm Whatman syringe filter and lyophilized to give a white solid, 190 (73 mg, 91%).

[0368] 9. Azide introduction Scheme 10-9 [ka] The following reaction conditions were used in the reaction shown in Scheme 10-9: a) 1-(azidosulfonyl)-1H-imidazol-3-ium chloride, copper(II) sulfate, potassium carbonate.

[0369] More specifically, 190 (72 mg, 0.13 mmol), 1-(azidosulfonyl)-1H-imidazol-3-ium chloride (80 mg, 0.38 mmol), copper sulfate (10 µmol), and potassium carbonate (79 mg, 0.57 mmol, 579 µL of a 1 M stock solution) were dissolved in water (1.96 mL). The reaction was mixed at room temperature and monitored by TLC (7:3:2 EtOH:NH4OH:HO). When all starting material was consumed, Cuprisorb™ was added to the reaction, which was incubated at room temperature for 2 hours to remove the copper from the solution. The resin was removed by filtration, the reaction volume was reduced under vacuum, and the product was purified by P2 size exclusion (70 x 2.5 cm column, 100 mM ammonium bicarbonate, 8 min / fraction, approximately 5 mL / fraction). The product-containing fractions (26-32) were pooled and dried under reduced pressure to give a white solid 200 (67 mg, 89% yield). 1H(600MHz,CDCl3),δ(ppm):GlcNAc:4.39(H1),3.59(H2),3.64(H5),4.26(ddd,J=10.9Hz,H6a), 4.17(ddd,J=11.0,3.7Hz,H6b);Gal:4.41(H1),3.38(H2),3.53(H3),3.77(H4),3.60(H6a and H6b) From HSQC 13 C(75MHz,CDCl3),δ(ppm):GlcNAc:102.5(C1),55.1(C2),65.9(C6),21.9(NHAc);Gal:101.3(C1),71.1(C2),72.1(C3),68.7(C4),61.0(C6)

[0370] ESI-TOF MS C 19 H 33 N4O 14 S(MH) - m / z calculated for = 573.1719, observed = 573.0749

[0371] 10. Chemical Synthesis of 6-Sulfo-N-acetylglucosamine Scheme 10-10 [ka] The reactions shown in Scheme 10-10 used the following conditions: a) AcO, sodium acetate; b) TMSOTf; c) 5-azidopentan-1-ol, TMSOTf; d) sodium methoxide; e) sulfur trioxide-pyridine. Note that Scheme 10-10 shows the same transformation as Scheme 1-6 in Procedure A. The numbering of the compounds in Scheme 10-10 is different from Scheme 1-6, but the compounds are the same.

[0372] More specifically, N-acetylglucosamine (100 g, 450 mmol) was added portionwise to a refluxing solution of acetic anhydride (340 mL, 3375 mmol) and sodium acetate (37.2 g, 450 mmol). The mixture was stirred at reflux until all materials dissolved, and TLC analysis (100% EtOAc, Rf = 0.47) indicated the reaction was complete. Excess acetic anhydride was removed under reduced pressure, and the crude material was dissolved in EtOAc (1 L), washed with saturated sodium bicarbonate (10 × 150 mL), brine (3 × 100 mL), dried over MgSO4, and reduced in vacuo to provide peracetylated N-acetylglucosamine (210, 129 g, 74%) as a white solid. 210 (5.4 g, 13.9 mmol) was dissolved in DCE (69 mL), and to this solution was added TMSOTf (2.5 mL, 13.87 mmol). The solution was warmed to 60 °C to promote oxazolidone formation, which was monitored by TLC (40% acetone / hex, Rf = 0.3). The reaction was quenched with TEA (2.2 mL, 15.18 mmol), the solvent was reduced, and the product was purified by flash chromatography (40% acetone / hex) to give 220 (3.9 g, 85%) as a clear oil. Tri-acetylated oxazolidone 220 (4.36 g, 13.24 mmol) was dissolved in DCE (66 mL). To this solution was added 5-azidopentan-1-ol (3.4 g, 26.48 mmol) and 4Å MS (3 g). The combination was mixed at room temperature, followed by the addition of TMSOTf (2.4 mL, 13.24 mmol). The reaction was warmed to 60 °C to promote glycosylation, and progress was monitored by TLC (90% EtOAc / Hex, Rf = 0.55). When all starting material was converted to product, the reaction was quenched with TEA (1.84 mL, 13.24 mmol) and cooled to room temperature. The solvent was removed under reduced pressure, and the glycosylated N-acetylglucosamine was recovered by flash chromatography (80→90% EtOAc / Hex) to provide 230 (5 g, 80%) as a white solid. 230 (14.38 g, 31.37 mmol) was dissolved in MeOH (156 mL), and the solution was adjusted to pH 9 with sodium methoxide in methanol (1 M stock solution).The reaction was monitored by TLC (20% MeOH / DCM, Rr=0.76) and when complete the pH was adjusted to 100 with Dowex 50WX8 H. + The mixture was neutralized with resin. The resin was removed by filtration, and the MeOH was removed under reduced pressure to give triol 240 (9.3 g, 90%) as a white solid. 6-O-Sulfation was achieved by dissolving 240 (600 mg, 1.8 mmol) in DMF (36 mL) and placing the mixture in an ice bath. To the cooled solution was added sulfur trioxide-pyridine complex (287 mg, 1.8 mmol). The reaction was stirred overnight at 4 °C for 3 days, with additional sulfur trioxide-pyridine (287 mg, 1.8 mmol) added daily. The reaction was monitored by TLC (20% MeOH / DCM, Rf = 0.23) and quenched with an aliquot of NaOH (5 M, pH → 8). The solvent was removed, and the reaction was purified by Bio-Gel size-exclusion chromatography. The product-containing fractions were lyophilized to give 250 (514 mg, 69%) as a white powder.

[0373] 2D of compounds 240 and 250 in Scheme 10-10 1 The H-NMR spectra are shown in Figures 1A and 1B.

[0374] 11. Enzymatic introduction of β1,4-Gal to form 6-sulfo-N-acetyllactosamine Schemes 10-11 (Same transformation as Schemes 1-7 in Step A - numbering is different) [ka] β1,4-Gal incorporation was achieved by dissolving 250 (5 mg, 11.7 μmol) and UDP-galactose (10 mg, 17.6 μmol) in Tris buffer (585 μL, pH 7.3, 0.1 M) containing 10 mM MnCl2. To this solution was added B4GALT1 (50 μg, 1% wt / wt) and calf intestinal alkaline phosphatase (CIAP, 5.85 μL, 1 kU stock). The reaction was incubated at 37°C for 3 h. The reaction progress was monitored by LC-ESI-MS equipped with a Waters XBridge BEH amide column, 2.5 μm, 130 Å, 2.1 × 150 mm (flow rate 0.25 mL / min, 10 mM ammonium formate, B = ACN, linear gradient 80% → 60% B over 18 min).

[0375] The reaction was monitored by following the crude reaction using hydrophilic interaction liquid chromatography / electrospray ionization mass spectrometry (HILIC-LC / ESI-MS). The results of such a follow-up are shown in Figure 2. If the reaction did not go to completion, excess UDP-Gal (5 mg, 5.85 μmol) and B4GALT1 (25 μg) were added to the reaction. The reaction mixture was filtered through a PALL Nanosep® centrifugal spin filter (3k MWCO), and the filtrate was purified by Bio-Gel P2 size-exclusion chromatography. The product-containing fractions were lyophilized to provide the desired disaccharide 200 (4.8 mg, 70%).

[0376] 12. Chemoenzymatic synthesis of CMP-Neu5-glycine Schemes 10-12 [ka] The reactions shown in Schemes 10-12 used the following conditions: a) sodium pyruvate, cytidine triphosphate disodium salt; PmNanA; NmCSS, PmPPA; b) Lindlar catalyst, H2.

[0377] More specifically, ManAz (260, 100 mg, 0.38 mmol), cytidine triphosphate disodium (219 mg, 0.42 mmol), and sodium pyruvate (210 mg, 1.91 mmol) were dissolved in Tris buffer (100 mM, pH 8.5, 19 mL) containing MgCl (10 mM).This reaction was carried out using 1 mg of Pasteurella multocida sialic acid aldolase (PmNanA) (Li, Y.; Yu, H.; Cao, H.; Lau, K.; Muthana, S.; Tiwari, VK; Son, B.; Chen, X., Pasteurella multocida sialic acid aldolase: a promising biocatalyst. Appl Microbiol Biot 2008, 79(6), 963), and 1 mg of Neisseria meningitidis group B CMP-sialic acid synthetase (NmCSS) (Yu, H.; Yu, H.; Karpel, R.; Chen, X., Chemoenzymatic synthesis of CMP-sialic acid derivatives by a one-pot two-enzyme system: comparison of substrate flexibility of three microbial CMP-sialic acid derivatives). synthetases. Bioorg Med Chem 2004,12 (24), 6427-6435) and inorganic pyrophosphatase from Pasteurella multocida (PmPPA) (1 mg) (Lau, K.; Thon, V.; Yu, H.; Ding, L.; Chen, Y.; Muthana, M. M.; Wong, D.; Huang, R.; Chen, X., Highly efficient chemoenzymatic synthesis of [small beta]1-4-linked galactosides with promiscuous bacterial [small beta]1-4-galactosyltransferases. Chem Commun 2010,46(33), 6066-6068) were added, and the reaction was incubated overnight at 37°C. When all of the 260 had been consumed as determined by TLC, calf intestinal alkaline phosphate (CIAP, 1000 units) was added to the reaction and the mixture was again incubated at 37°C overnight.On day 3, cold ethanol (17 mL) was added to the mixture, and all insoluble material was removed by centrifugation. The supernatant was concentrated, and CMP-Neu5Az (270) was purified by P2 size-exclusion chromatography (5 × 60 cm, elution with 100 mM ammonium bicarbonate, 15 min / fraction or approximately 35 mL). Product-positive fractions were pooled to give a white fluffy solid (185 mg, 74%), and analytical details were consistent with those found in the literature. 270 (185 mg, 0.28 mmol) was dissolved in 20 mM ammonium hydroxide (28 mL), and Lindlar's catalyst (9 mg) was added to the mixture. The mixture was stirred under a hydrogen atmosphere until no starting material was observed by TLC (7:3:2 EtOH:NH4OH:HO). The catalyst was removed with a 0.2 μm syringe filter and the filtrate was dried by lyophilization to give a white solid 280 (219 mg, 92%) which was used without further purification.

[0378] Compound 280 was analyzed by LC-MS using a Waters XBridge BEH, amide column, 2.5 μm, 130 Å, 2.1 × 150 mm, with an ESI detector at a flow rate of 0.25 mL / min; 80% to 60% in 18 min (A = 10 mM ammonium formate, pH 3.4, B = acetonitrile). The peak for compound 280 eluted at 19.0 min. Electrospray ionization time-of-flight mass spectrometry (ESI-TOF MS) was used to determine the molecular mass of compound 280; C 20 H 31 N5O 16 P(MH) - m / z calculated for = 628.1509, found = 628.0397.

[0379] 13. Enzymatic incorporation of α2,3-Neu5-glycine Schemes 10-13 [ka] The following reaction conditions were used in the reactions shown in Schemes 10-13: a) PmST1, CIAP.

[0380] Specifically, 200 (70 mg, 0.12 mmol) and 280 (115 mg, 183 mmol) were dissolved in 12 mL of Tris buffer (100 mM, pH 8.5) containing 700 μg of Pasteurella multocida sialyltransferase-I (PmSt1) (Yu H, Chokhawala H, Karpel R, Yu H, Wu B, Zhang J, Zhang Y, Jia Q, Chen XA. Multifunctional Pasteurella multocida sialyltransferase: A powerful tool for the synthesis of sialoside libraries. J Am Chem Soc. 2005;127:17618-17619) and CIAP (120 U). The reaction was incubated at 30 °C until all 200 was consumed, as determined by TLC (7:3:2 EtOH:NH4OH:HO). After all the starting disaccharide was consumed, cold EtOH was added to the mixture (12 mL) and insoluble material was removed by centrifugation. The supernatant was concentrated and purified by P2 size-exclusion chromatography (2.5 × 70 cm, elution with 100 mM ammonium bicarbonate, 8 min / fraction). Fractions containing the desired product were pooled and concentrated by lyophilization to give 290 as a white solid (82 mg, 77% yield).

[0381] Compound 290 was analyzed by LC-MS HPLC using a Waters XBridge BEH, amide column, 2.5 μm, 130 Å, 2.1 × 150 mm, with ESI as the detector, at a flow rate of 0.25 mL / min; 80% → 60% A in 18 min (A = 10 mM ammonium formate, pH 3.4, B = acetonitrile). The peak for compound 290 eluted at 11.2 min. The molecular mass of compound 290 was determined using ESI-TOF MS: C 30 H 51 NO 22 S(MH) - m / z calculated for = 879.2783, found = 879.1189.

[0382] 14. Enzymatic introduction of α1,3-fucose using FUT6 Schemes 10-14 [ka] The following conditions were used in the reactions shown in Schemes 10-14: a) FUT6, GDP-fucose.

[0383] More generally, 290 (71 mg, 80.8 μmol) and GDP-fucose (102 mg, 161 μmol) were dissolved in 4 mL of sodium cacodylate buffer (100 mM, pH 6.5) containing 10 mM MnCl2. To this solution was added FUT6 (50 μg) and CIAP (40 units). The reaction was incubated overnight at 37 °C, and the next day progress was monitored by a Shimadzu LC-ESI-IT-TOF with a Waters XBridge BEH, amide column, 2.5 μm, 130 Å, 2.1 × 150 mm, using ESI as the detector, at a flow rate of 0.25 mL / min; 20% → 40% A in 18 min (A = 10 mM ammonium formate, pH 3.4, B = acetonitrile). If the reaction did not go to completion, additional aliquots of FUT6 were added until the reaction reached completion. Upon completion, cold EtOH (4 mL) was added and insoluble material was removed by centrifugation. The product was purified by P2 chromatography (70 × 2.5 cm, 100 mM ammonium bicarbonate, 8 min / fraction). Fractions containing the product were pooled to give 300 (65 mg, 79%) as a white fluffy solid. 1H(600MHz,CDCl3),δ(ppm):GlcNAc:4.42(d,J=7.9Hz,H1),3.75(H2),3.73(H3),3.86(H4),3.64(H5),4.23( H6a and H6b);Gal:4.47(d,J=7.8Hz,H1),3.36(H2),3.97(dd,J=9.9,3.1Hz,H2),3.82(H3),3.46(H4),3.55(H 6a and H6b); Fuc: 4.96 (d, J = 3.9 Hz, H1), 3.52 (H2), 3.75 (H3), 3.64 (H4), 4.68 (H5), 1.04 (d, J = 6.6 Hz, CH3); Neu5 Glycine: 2.61 (dd, J = 12.4, 6.0 Hz, H3-equatorial), 1.68 (dd, J = 12.2 Hz, H3-axial), 3.57 (H4), 3.75 (H9a and H9b). From HSQC 13 C(75MHz,CDCl3),δ(ppm):GlcNAc:100.9(C1),55.8(C2),74.7(C3),73.0(C4),72.9(C5),65.9(C6);Gal:101.3(C1),69 .6(C2),75.4(C3),67.2(C4),67.9(C5),61.3(C6);Fuc:98.8(C1),67.0(C2),69.2(C3),71.8(C4),66.9(C5)15.9(CH3); Neu5 Glycine: 39.5 (C3), 72.6 (C4), 51.7 (C5), 62.7 (C9)

[0384] Compound 300 was analyzed by HPLC LC-MS using a Waters XBridge BEH, amide column, 2.5 μm, 130 Å, 2.1 × 150 mm, with ESI as the detector, at a flow rate of 0.25 mL / min; 80% to 60% in 18 min (A = 10 mM ammonium formate, pH 3.4, B = acetonitrile). The peak for compound 300 eluted at 13 min. The molecular mass of compound 300 was determined using ESI-TOF MS: C 36 H 61 NO 26 S(MH) - m / z calculated for = 1025.3362 found = 1025.1684.

[0385] 15. Enzymatic synthesis of 9-aminoCMP-Neu5Ac Schemes 10-15 [ka] The following conditions can be used in the reactions shown in Schemes 10-15: a) sodium pyruvate, cytidine triphosphate disodium salt; PmNanA; NmCSS, PmPPA; b) Lindlar catalyst, H2.

[0386] More specifically, 330 can be synthesized following the same protocol as shown for the synthesis of 280.

[0387] 16. Enzymatic introduction of α2,3-Neu5Ac-9-amino Schemes 10-16 [ka] The following conditions can be used in the reactions shown in Schemes 1-16: a) PmST1, CIAP.

[0388] More specifically, 340 can be synthesized following the same protocol as shown for the synthesis of 290.

[0389] 17.Enzymatic introduction of α1,3-fucose using FUT6 Schemes 10-17 [ka] The following conditions can be used in the reactions shown in Schemes 10-17: a) FUT6, GDP-fucose.

[0390] More specifically, 350 can be synthesized following the same protocol as for the synthesis of 300.

[0391] II. Library of S-compounds by procedure B Exemplary S-compounds were prepared according to procedure B illustrated below.

[0392] Synthesis of S106-azide Schemes 10-18 [ka] The following reaction conditions were used in the reactions shown in Schemes 10-18: a) Triethylamine.

[0393] More specifically, 300 (14 mg, 13.66 μmol) and quinoline-2-carbonyl-NHS ester (11 mg, 54.6 μmol) were dissolved in a 1:1 mixture of HO / DMF (683 μL), and triethylamine (11 μL, 81.9 μmol) was added to the solution. The reaction mixture was incubated overnight at 37 °C, and the next day, progress was monitored by LC-ESI-MS equipped with a Shimadzu C18 5 μm, 50 × 4.6 mm column, with a flow rate of 0.25 mL / min and a 10-min gradient of 0 to 100% acetonitrile (solvent A = 10 mM ammonium formate, pH 3.4). When all starting material was consumed, the product was purified by P2 chromatography (1.5 × 80 cm, 100 mM ammonium bicarbonate, 15 drops / fraction), and the product-containing fractions were pooled and lyophilized to give S106-azide (11 mg, 68%) as a white fluffy solid.

[0394] An LC-MS trace of S106-azide was performed using the same conditions described above to monitor the progress of the reaction. The product, S106-azide, eluted at 6 minutes. The molecular mass of compound S106-azide was determined using ESI-TOF MS: 46 H 66 N7O 27 S(MH - ) m / z calculated = 1180.3733 found = 1180.1682.

[0395] The HSQC (600 MHz) 2D NMR spectrum of S106 azide is shown in FIG.

[0396] The following representative compounds were made according to Scheme 10 in general, and Schemes 10-18 in particular. All compounds were analyzed by LC-ESI-MS equipped with a Shimadzu C18 5 μm, 50 × 4.6 mm column at a flow rate of 0.25 mL / min, with a gradient of 0 to 100% acetonitrile in 10 min (solvent A = 10 mM ammonium formate, pH 3.4).

[0397] S101-Azide: [ka] S101-azide: Product eluted at 6.75 min. The molecular mass of S101-azide was determined using ESI-TOF MS: C 47 H 67 NO 27 S(MH - ) calculated m / z = 1179.3780, found = 1179.1732.

[0398] S103-Azide: [ka] S103-azide: Product eluted at 5.6 min. The molecular mass of S103-azide was determined using ESI-TOF MS: C 41 H 63 NO 27 S2(MH - ) calculated m / z = 1135.3188, found = 1135.1208.

[0399] S105-Azide: [ka] S105-azide: Product eluted at 6.5 min. The molecular mass of S105-azide was determined using ESI-TOF MS: C 44 H 64 N7O 27 S2(MH - ) calculated m / z = 1186.3297 found = 1186.0807.

[0400] S107-Azide [ka] S107-azide: Product eluted at 7 min. The molecular mass of S107-azide was determined using ESI-TOF MS: C 49 H 69 NO 27 S(MH - ) calculated m / z = 1205.3937, found = 1205.1807.

[0401] S110-Azide: [ka] The product eluted at 6.6 min. The molecular mass of S110-azide was determined using ESI-TOF MS: C 45 H 65 NO 27 S2(MH - ) calculated m / z = 1185.3345, found = 1185.1246.

[0402] S112-Azide: [ka] The molecular mass of S112-azide was determined using ESI-TOF MS:C 45 H 64 ClNO 27 S2(MH - ) calculated m / z = 1219.2955, found = 1219.1099.

[0403] Example 2: Synthesis of a library of D-compounds The following reaction scheme was used for the synthesis of the D series of compounds.

[0404] Scheme 2: Attachment of terminally modified fragments [ka] In Scheme 2, the notation D1XX denotes a compound in which the N-acetyl at position 5 of the terminal sialic acid is substituted with one of the fragments described below, and the notation D2XX denotes a compound in which the N-acetyl at position 9 of the terminal sialic acid is substituted with one of the fragments described below.

[0405] I. Synthesis of precursors 45 and 46 The precursor compounds 45 and 46 in Scheme 2 were synthesized as follows.

[0406] 1. Chemical synthesis of anomeric lactose Scheme 1-2 [ka] The following conditions were used in the reactions shown in Scheme 1-2: a) BzCl, pyridine; b) HBr-AcOH / Ac2O; c) 5-azidopentan-1ol, AgOTf; d) sodium methoxide.

[0407] More specifically, lactose (25 g, 73 mmol) was suspended in pyridine (300 mL) and the mixture was placed in an ice bath. To the cooled solution, benzoyl chloride (100 mL, 876 mmol) was added dropwise over 30 min. After all the benzoyl chloride was added, the reaction mixture was brought to room temperature and vigorously stirred until the lactose was perbenzoylated, as determined by TLC (30% EtOAc / Hex, Rf = 0.30). The contents of the reaction mixture were added to an ice / water mixture, which resulted in the formation of an oily layer after standing. The aqueous phase was decanted, leaving behind a bottom oily material, which was dissolved in boiling EtOH and left at -20 °C overnight to give product 36 as a white / yellow solid, which was carried forward without further purification. The perbenzylated lactose was suspended in acetic acid (142 mL) and acetic anhydride (47 mL) in an ice bath. To this suspension, HBr-AcOH (176 mL, 10 eq) was added dropwise over 1 h. After all the HBr-AcOH was added, the reaction was allowed to reach room temperature and stirred vigorously. The reaction progress was monitored by TLC (30% EtOAc / Hex, Rf = 0.44). The reaction was worked up by dilution in EtOAc (2 L), the acid extracted with water (5 × 250 mL), sodium bicarbonate (the reaction was added to a beaker, and the stirring solution was added to the solid sodium bicarbonate), quenched with brine (2 × 200 mL), dried over MgSO4, and reduced to give the anomeric bromide product (3, 74.9 g, 91% 2 steps) as a white solid. 37 (75 g, 66 mmol) was dissolved in toluene (660 mL) containing 20 g of 4Å MS and 5-azidopentan-1-ol (9.3 g, 72 mmol). The mixture was stirred at room temperature for 30 min and then cooled to −80 °C in a dry ice bath. To the cooled reaction was added silver trifluoromethanesulfonate (21 g, 82.5 mmol). The reaction was monitored by TLC (30% EtOAc / Hex, Rf = 0.34), and upon completion, the reaction mixture was filtered over a pad of Celite, reduced, and purified by flash chromatography (30% EtOAc / Hex) to give 38 (53.8 g, 69%) as a white solid. 38 (53.8 g, 45.5 mmol) was dissolved in MeOH (455 mL) and the pH was adjusted to 9 by the addition of a 1 M solution of sodium methoxide.The reaction was monitored by TLC (20% MeOH / DCM, Rf=0.57) and purified using Dowex 50WX8 H. + Quenching with resin (pH adjusted to neutral) and reducing the solvent gave a brown solid which was washed with acetone (acetone removed the benzyl methyl ester) to give a light brown solid (39, 17.3 g, 84%).

[0408] 2. Chemoenzymatic synthesis of Fmoc-protected alpha 2,3-sialyllactose Scheme 2-2 [ka] The following conditions were used in the reaction shown in Scheme 2-2: a) PmST1, NmCSS, PmPPA, Neu5Ac, CTP; b) Pd / C, H2, c) Fmoc-OSu.

[0409] More specifically, lactose 39 (100 mg, 0.22 mmol) containing an anomeric linker was dissolved in Tris buffer (11 mL, 100 mM, pH 8.5) containing MgCl (10 mM), N-acetylneuraminic acid (102 mg, 0.33 mmol), and cytidine-5'-triphosphate (159 mg, 0.33 mmol). The solution was adjusted to pH 8.5 using 5 M NaOH. To this solution were added PmST1 (1 mg), NmCSS (2 mg), and PmPPA (2 mg). The reaction mixture was incubated at 37 °C with periodic inversion of the reaction vessel. Progress was monitored by TLC (7:3:2 EtOH, NH4OH, HO), and when complete conversion was observed, the enzyme was removed by precipitation facilitated by the addition of EtOH (11 mL). The mixture was placed at -20°C for 2 hours, and insoluble material was removed by centrifugation. The crude product was purified using Bio-Gel P2 size-exclusion chromatography. The product-containing fractions were concentrated and lyophilized to give the trisaccharide 40 (142 mg, 87%) as a white, fluffy solid. 40 (142 mg, 0.19 mmol) was dissolved in t-BuOH / HO (1.9 mL, 1:1 v / v) containing Pd / C (7 mg). The reactants were mixed under a hydrogen atmosphere at room temperature, and the reaction progress was monitored by ESI-MS. The catalyst was removed using a 0.2 μm PALL Acrodisc syringe filter, and the solution was lyophilized to yield a white, fluffy solid (41). 41 was dissolved in DI water (9.5 mL) containing sodium bicarbonate (4.5 mmol). In a separate vial, Fmoc-OSu (128 mg, 0.38 mmol) was dissolved in ACN (9.5 mL), and 41 was added dropwise to this vial with vigorous mixing. The reaction progress was monitored by ESI-MS. After all starting material was converted to the desired product, the reaction was reduced in vacuo and purified by preparative HPLC (Agilent 1200, Eclipse XDB-C18, 21.2 × 250 mm, 7 μm column; A solvent = 50 mM ammonium bicarbonate; B = ACN, λ = 262 μm) using a linear gradient of 10 to 60% over 30 min at a flow rate of 20 mL / min. The compound-containing fractions were lyophilized to give the Fmoc-protected trisaccharide 42 (100 mg, 56% over two steps) as a white fluffy solid.

[0410] 3. Enzymatic synthesis of GD3-azide derivatives Scheme 2-3 [ka] The following conditions were used in the reactions shown in Scheme 2-3: a) CSTII, CMP-Neu5Ac-9-N3; b) CSTII, CMP-Neu5Az.

[0411] More specifically, 42 (50 mg, 53 μmol) was treated with MgCl (10 mM), CMP-Neu5Ac-9-N3 or CMP-Neu5Az (106 μmol), CSTIIΔ32 I53S (1 mg) and CIAP (53 μL, 1 kU stock solution) 6 The reaction mixture was dissolved in Tris buffer (5.3 mL, 100 mM, pH 8.5) containing 43 (40 mg, 59%). The reaction mixture was incubated at 37 °C and monitored by ESI-MS. When no further product conversion was observed, the reaction mixture was filtered through a PALL Nanosep® centrifugal spin filter (3 kJ MWCO), and the filtrate was purified by Bio-Gel P2 size-exclusion chromatography. The product-containing fractions were lyophilized to afford the desired tetrasaccharides 43 (40 mg, 59%) and 44 (44 mg, 65%) as white fluffy solids.

[0412] 4. General Protocol for α2,8-Azido-Sialic Acid Reduction Scheme 2-4 [ka] The following reaction conditions were used in the reactions shown in Schemes 2-4: Zn, AcOH, H2O.

[0413] More specifically, 44 (30 mg, 23.5 μmol) was dissolved in HO (3 mL), and to this solution was added acid-treated Zn(s) (60 mg). The mixture was placed in an ice bath, and acetic acid (100 μL) was added to the cooled solution. The reaction was monitored by ESI-MS, and when all starting material was converted to product, the reaction was quenched with saturated ammonium bicarbonate (pH 7). The reaction was filtered through a 0.2 μm PALL Acrodisc syringe filter and purified by preparative HPLC (Agilent 1200, Eclipse XDB-C18, 21.2 × 250 mm, 7 μm column) using a linear gradient of 10 to 60% B in 30 min at a flow rate of 20 mL / min. A solvent = 50 mM ammonium bicarbonate; B = ACN, λ = 262 μm. The compound-containing fractions were lyophilized to give the Fmoc-protected trisaccharide 45 (20 mg, 69%) as a white fluffy solid.

[0414] 5. General Protocol for Fmoc Deprotection Deprotection of D series compounds was generally carried out according to Schemes 1-13 above.

[0415] II.D - Compound Library The tetrasaccharide (45 or 46, 1 mg, approximately 0.8 μmol) was dissolved in DMF (100 μL) containing diisopropylethylamine (DIPEA, 4 μmol). To this solution was added acyl chloride (1.6 μmol), and the reaction was mixed using vortexing for 30 minutes. The reaction was monitored by ESI, and upon completion, conversion to the product was achieved. The product was purified by HPLC chromatography under the following conditions: Agilent 1200 HPLC, Eclipse XDB-C8, 4.6 × 250 mm, 5 μm analytical column, solvent = 50 mM ammonium bicarbonate; B = ACN, λ = 262 μm), linear gradient from 10 to 60% B in 30 minutes, flow rate 1 mL / min. Fractions containing the desired product were lyophilized to give the desired product as a white fluffy solid.

[0416] The following compounds of the D series were prepared using the terminally modified fragments listed in Table 1 above. The identity of the compounds is as follows: 1The R of the D series compounds listed below was confirmed using H-NMR spectroscopy. [ka] is.

[0417] Compound D101 (0.15 μmol) was prepared using the general procedure. [ka]

[0418] Compound D102 (0.15 μmol) was prepared using the general procedure. [ka]

[0419] Compound D103 (0.15 μmol) was prepared using the general procedure. [ka]

[0420] Compound D104 (0.15 μmol) was prepared using the general procedure. [ka]

[0421] Compound D105 (0.15 μmol) was prepared using the general procedure. [ka]

[0422] Compound D106 (0.15 μmol) was prepared using the general procedure. [ka]

[0423] Compound D107 (0.15 μmol) was prepared using the general procedure. [ka]

[0424] Compound D108 (0.15 μmol) was prepared using the general procedure. [ka]

[0425] Compound D109 (0.15 μmol) was prepared using the general procedure. [ka]

[0426] Compound D110 (0.15 μmol) was prepared using the general procedure. [ka]

[0427] Compound D111 (0.15 μmol) was prepared using the general procedure. [ka]

[0428] Compound D112 (0.15 μmol) was prepared using the general procedure. [ka]

[0429] Compound D113 (0.15 μmol) was prepared using the general procedure. [ka]

[0430] Compound D114 (0.15 μmol) was prepared using the general procedure. [ka]

[0431] Compound D115 (0.15 μmol) was prepared using the general procedure. [ka]

[0432] Compound D116 (0.15 μmol) was prepared using the general procedure. [ka]

[0433] Compound D117 (0.15 μmol) was prepared using the general procedure. [ka]

[0434] Compound D118 (0.15 μmol) was prepared using the general procedure. [ka]

[0435] Compound D119 (0.15 μmol) was prepared using the general procedure. [ka]

[0436] Compound D120 (0.15 μmol) was prepared using the general procedure. [ka]

[0437] Compound D121 (0.15 μmol) was prepared using the general procedure. [ka]

[0438] Compound D122 (0.15 μmol) was prepared using the general procedure. [ka]

[0439] Compound D123 (0.15 μmol) was prepared using the general procedure. [ka]

[0440] Compound D124 (0.15 μmol) was prepared using the general procedure. [ka]

[0441] Compound D125 (0.15 μmol) was prepared using the general procedure. [ka]

[0442] Compound D126 (0.15 μmol) was prepared using the general procedure. [ka]

[0443] Compound D127 (0.15 μmol) was prepared using the general procedure. [ka]

[0444] Compound D128 (0.15 μmol) was prepared using the general procedure. [ka]

[0445] Compound D129 (0.15 μmol) was prepared using the general procedure. [ka]

[0446] Compound D130 (0.15 μmol) was prepared using the general procedure. [ka]

[0447] Compound D131 (0.15 μmol) was prepared using the general procedure. [ka]

[0448] Compound D201 (0.15 μmol) was prepared using the general procedure. [ka]

[0449] Compound D202 (0.15 μmol) was prepared using the general procedure. [ka]

[0450] Compound D203 (0.15 μmol) was prepared using the general procedure. [ka]

[0451] Compound D204 (0.15 μmol) was prepared using the general procedure. [ka]

[0452] Compound D205 (0.15 μmol) was prepared using the general procedure. [ka]

[0453] Compound D206 (0.15 μmol) was prepared using the general procedure. [ka]

[0454] Compound D207 (0.15 μmol) was prepared using the general procedure. [ka]

[0455] Compound D208 (0.15 μmol) was prepared using the general procedure. [ka]

[0456] Compound D209 (0.15 μmol) was prepared using the general procedure. [ka]

[0457] Compound D210 (0.15 μmol) was prepared using the general procedure. [ka]

[0458] Compound D211 (0.15 μmol) was prepared using the general procedure. [ka]

[0459] Compound D212 (0.15 μmol) was prepared using the general procedure. [ka]

[0460] Compound D213 (0.15 μmol) was prepared using the general procedure. [ka]

[0461] Compound D214 (0.15 μmol) was prepared using the general procedure. [ka]

[0462] Compound D215 (0.15 μmol) was prepared using the general procedure. [ka]

[0463] Compound D216 (0.15 μmol) was prepared using the general procedure. [ka]

[0464] Compound D217 (0.15 μmol) was prepared using the general procedure. [ka]

[0465] Compound D218 (0.15 μmol) was prepared using the general procedure. [ka]

[0466] Compound D219 (0.15 μmol) was prepared using the general procedure. [ka]

[0467] Compound D220 (0.15 μmol) was prepared using the general procedure. [ka]

[0468] Compound D221 (0.15 μmol) was prepared using the general procedure. [ka]

[0469] Compound D222 (0.15 μmol) was prepared using the general procedure. [ka]

[0470] Compound D223 (0.15 μmol) was prepared using the general procedure. [ka]

[0471] Compound D224 (0.15 μmol) was prepared using the general procedure. [ka]

[0472] Compound D225 (0.15 μmol) was prepared using the general procedure. [ka]

[0473] Compound D226 (0.15 μmol) was prepared using the general procedure. [ka]

[0474] Compound D227 (0.15 μmol) was prepared using the general procedure. [ka]

[0475] Compound D228 (0.15 μmol) was prepared using the general procedure. [ka]

[0476] Compound D229 (0.15 μmol) was prepared using the general procedure. [ka]

[0477] Compound D230 (0.15 μmol) was prepared using the general procedure. [ka]

[0478] Compound D231 (0.15 μmol) was prepared using the general procedure. [ka]

[0479] Example 3: Conjugation of polymers and glycans The following synthetic schemes were used to conjugate glycans with polymers. In these schemes, the glycans are partially represented using only carbohydrate units directly attached to linker groups. Additional preparation protocols are provided in Example 8.

[0480] Synthesis Example 3-1: Preparation of glycans with click reagents Synthetic Scheme 3-1 [ka] Synthetic Scheme 3-2 [ka] Synthetic Scheme 3-3 [ka] Synthetic Scheme 3-4 [ka] Synthetic Schemes 3-5 [ka] Synthetic Schemes 3-6 [ka] For each of Synthetic Schemes 3-1 through 3-6, the anomeric linker is modified with the corresponding click reagent using the following general protocol: Dissolve amine-terminated glycan (1 eq.) and click reagent (5 eq.) in DMF to a final glycan concentration of 10 mM. Add diisopropylethylamine (DIPEA, 5 eq.) to this solution and incubate the reaction at 37 °C until no further starting material is observed by ESI-MS. Purify the reaction by P-2 size-exclusion chromatography eluting with 0.1 M ammonium bicarbonate. Pool product-containing fractions and remove the solvent by lyophilization.

[0481] Those skilled in the art will appreciate that many other click chemistry reagents are commercially available and can be used depending on the need, availability, and desirability of the final product. For example, with respect to Synthetic Scheme 3-1, any one of the following commercially available click reagents (from vendors such as Broadpharma, Sigma-Millipore, or Fisher Scientific) can be used: [ka]

[0482] Synthesis Example 3-2: Exemplary nanoparticle production using polymers with DBCO. Nanoparticles are prepared by the emulsion method. A representative nanoparticle preparation example involves the following: PLGA(10k)-PEG(5k)-COOH and PLGA(10k)-PEG(5k)-DBCO are weighed in a 3:1 weight ratio and dissolved in a mixture of organic solvents. The polymer concentration is 37 mg / mL in organic solvent (59.5% ethyl acetate, 40.5% benzyl alcohol). 10 mL of the polymer solution is added to 40 mL of an aqueous phase consisting of 0 or 0.1% Tween 80 in water and homogenized using a rotatator to form a coarse emulsion. The coarse emulsion is further homogenized into a fine nanoemulsion by passing it three times using a microfluidizer. The nanoemulsion is then quenched by adding it to 450 mL of cold water to harden the nanoparticles. The quenched nanoparticle solution is then concentrated and washed by tangential flow filtration to remove the organic solvent. The concentration of the polymer nanoparticles is determined by evaporating water from a known volume of the nanoparticle solution, and the nanoparticle solution was then prepared for conjugation to oligoglycan ligands.

[0483] Synthesis Example 3-3: Ligand-azide conjugation to the surface of core DBCO nanoparticles via SPAAC chemistry The purified N3 ligand, prepared as described above, is conjugated to the surface of purified PLGA-DBCO nanoparticles using the SPAAC copper-free click chemistry protocol. Briefly, the DBCO-containing nanoparticles are incubated with the ligand-azide overnight at 4°C, room temperature, or 37°C. After the conjugation reaction is complete, the nanoparticles are washed using TFF to remove unreacted components, and the buffer is exchanged for an appropriate isotonic solution, such as 10% sucrose. The nanoparticle solution is then sterile-filtered through a 0.2 μm filter.

[0484] Synthesis Examples 3-4: Generation of Additional Exemplary Nanoparticles by Ligand-Azide Preconjugation to DBCO-Polymer Nanoparticles were prepared by the emulsion method. A representative example of nanoparticle production is described below. 10 mg of ligand-azide was dissolved in 0.6 mL of DMSO, DMF, acetonitrile, or some other suitable solvent. The dissolved ligand solution was added to 15 mg of PLGA(10k)-PEG(5k)-DBCO (dissolved in an organic solvent mixture of ethyl acetate and benzyl alcohol). The mixture was stirred and mixed overnight to conjugate the ligand-azide with PLGA-PEG-DBCO via azide-DBCO click chemistry coupling. After mixing overnight, 285 mg of PLGA(10k)-PEG(5k)-COOH was added to the reaction mixture and mixed / stirred until the solution was visually clear.

[0485] The final volume of organic solvent was 3.03 mL, containing 2.43 mL of ethyl acetate:benzyl alcohol (60:40 volume ratio) and 0.6 mL of DMSO. The polymer, fluorescein-PLGA, can be used to introduce fluorescent moieties into the nanoparticles. The total proportion of fluorescein polymer in the total polymer used to prepare the nanoparticles was 1 wt%. Thus, 3 mg of PLGA-fluorescein was also added to the polymer solution.

[0486] The polymer solution is added to 27 mL of cold water (saturated with ethyl acetate) and homogenized using an IKA T-18 Rotastator to form a coarse emulsion. The coarse emulsion is further homogenized into a fine nanoemulsion by passing it three times through a microfluidizer (Microfluidics LM10). The nanoemulsion is then quenched by adding it to 270 mL of cold water to harden the nanoparticles. The quenched nanoparticle solution is then concentrated and washed with cold water via tangential flow filtration (KR2i TFF, Repligen) to remove organic solvents and unreacted ligand-azide. The concentration of the polymer nanoparticles is determined by evaporating water from a known volume of nanoparticle solution. Sucrose is added to the nanoparticle solution at 10% wt / wt and filtered using a 0.2 μm Millipore syringe filter. The nanoparticle solution is frozen at -20°C. The size of the nanoparticles is measured using dynamic light scattering using a Malvern Zetasizer. Dynamic light scattering techniques utilize the constant, random thermal motion of particles and molecules, known as Brownian motion, to measure size. Particles diffuse at a rate related to their size, with smaller particles diffusing faster than larger ones. The diffusion rate is measured from the speckle pattern generated by illuminating the particles with a laser. Fluctuations in scattered intensity at specific angles are detected using a highly sensitive photodiode detector. The intensity changes are analyzed with a digital autocorrelator to generate a correlation function. This curve is then analyzed to obtain the particle size and size distribution. Nanoparticles are diluted with clean water to a concentration of approximately 0.1-1.0 mg / mL and measured with a Zetasizer. Each value obtained is the average of three readings.

[0487] Synthetic Examples 3-5: Conjugation of Glycans to Polymers by Amide Coupling Synthetic Schemes 3-7 [ka] Carboxylic acid-terminated PLGA or PLGA-PEG is dissolved in DCM to a final concentration of 5 mM. The solution is mixed at room temperature, and EDC and NHS or sulfo-NHS are added to the sample to bring the final concentration of each reagent to 25 mM. The solution is stirred at room temperature for 1 hour to obtain the desired NHS or sulfo-NHS activated ester. The activated ester is precipitated by the addition of MeOH (10x the reaction volume) and collected by centrifugation (2,000 g for 10 minutes). The activated PLGA-PEG-NHS pellet is redissolved in DCM and precipitated in MeOH a total of three times to ensure complete removal of residual EDC and NHS or sulfo-NHS. Amide formation is facilitated by dissolving the amine-containing glycan and activated PLGA in a mixture of organic solvents to promote reaction between the amine moiety of the glycan-amine and the NHS group on the PLGA-PEG-NHS. The choice of organic solvent can be optimized for each glycan-amine and PLGA-PEG-NHS system. For example, glycans are typically dissolved in DMSO or DMF, with a small amount of water added if necessary. The PLGA-PEG-NHS block copolymer is dissolved in DMSO or DMF. The glycan and PLGA-PEG-NHS solution should be clear and then mixed to initiate the reaction. N,N-diisopropylethylamine (DIPEA) is added to the reaction mixture to allow coupling of the PLGA-PEG and glycan. The reaction is carried out for at least 4 hours. After the reaction proceeds to completion, the organic solvent mixture is emulsified in an aqueous phase to obtain a nanoemulsion. Unreacted components of the reaction are expected to be removed during nanoparticle production. The aqueous phase used during nanoemulsion formation can be slightly basic, at pH 7.2 or higher, to aid in the hydrolysis of unreacted NHS groups to COOH.

[0488] Synthetic Examples 3-6: Conjugation of Glycans to Polymers via Azide / Alkyne Click Coupling Synthetic Schemes 3-8 [ka] Azide-containing glycans (>1.2 eq) and alkyne-containing polymers (1 eq) are mixed in a solvent system containing DMSO or DMF, benzyl alcohol, and ethyl acetate. The reaction is allowed to proceed to completion for at least 4 hours. Unreacted alkyne groups in the polymer can be capped by adding an excess of azide-containing molecules. After the reaction has proceeded to completion, the organic solvent mixture is emulsified in an aqueous phase to yield a nanoemulsion. Unreacted components of the reaction are expected to be removed during nanoparticle production.

[0489] Synthetic Examples 3-7: Conjugation of Glycans to Polymers by Copper-Free Click Coupling Synthetic Scheme 3-9A [ka] Synthetic Scheme 3-9B [ka] Synthetic Scheme 3-9C [ka] An azide-containing glycan (1 eq) and a cyclooctyne-containing polymer (1 eq) are mixed in a solvent system containing DMSO or DMF, benzyl alcohol, and ethyl acetate. The reaction is allowed to proceed to completion for at least 4 hours. Unreacted alkyne groups in the polymer can be capped by adding an excess of an azide-containing molecule. After the reaction proceeds to completion, the organic solvent mixture is emulsified in an aqueous phase to produce a nanoemulsion. Unreacted components of the reaction are expected to be removed during nanoparticle production.

[0490] Synthetic Scheme 3-9D [ka] Thiol-containing glycan (1 eq) and maleimide-containing polymer (1 eq) are mixed in a solvent system containing DMSO or DMF, benzyl alcohol, and ethyl acetate. The reaction is allowed to proceed to completion for at least 4 hours. Unreacted maleimide groups in the polymer can be capped by adding an excess of a thiol-containing molecule, such as cysteine. After the reaction proceeds to completion, the organic solvent mixture is emulsified in an aqueous phase to produce a nanoemulsion. Unreacted components of the reaction are expected to be removed during nanoparticle formation.

[0491] The above process for making nanoparticles can be applied to various combinations of coupling chemistries, including maleimide-thiol or cysteine, NHS and amine, click chemistry, etc.

[0492] Example 4: Microarray printing and screening The compounds described herein were first tested for binding to Siglec targets using the microarray printing technique described above.

[0493] All compounds were printed onto NHS-ester-activated glass slides (NEXTERION® Slide H, Schott Inc.) using a Scienion sciFLEXARRAYER S3 non-contact microarrayer equipped with a Scienion PDC80 nozzle (Scienion Inc.). Individual samples were dissolved in sodium phosphate buffer (50 μL, 0.225 M, pH 8.5) at a concentration of 100 μM and printed in 10 replicates with a spot volume of approximately 400 pL at 20°C and 50% humidity. Each slide contained 24 subarrays in a 3 × 8 layout. After printing, the slides were incubated in a humidity chamber for 8 h and then blocked with 5 mM ethanolamine in Tris buffer (pH 9.0, 50 mM) for 30 min at 40°C. The blocked slides were rinsed with DI water, spin-dried, and stored at room temperature in a desiccator for future use.

[0494] Screening was performed by incubating slides with protein solutions for a specified amount of time, followed by washing and drying. The buffers used for screening were TSM buffer (TSM, 20 mM Tris-Cl, pH 7.4, 150 mM NaCl, 2 mM CaCl, and 2 mM MgCl), TSM binding buffer (TSMBB, TSM buffer containing 0.05% Tween-20 and 1% BSA), and TSM wash buffer (TSMWB, TSM buffer containing 0.05% Tween-20). A typical washing procedure involved immersing glass slides in TSM wash buffer (2 min, containing 0.05% Tween-20), TSM buffer (2 min), and water (2 × 2 min), followed by spin drying.

[0495] For screening of Fc-tagged Siglec-2, -3, -5, -6, -7, and -9, slides were incubated with Siglec-2 (Fc-tagged, R&D system catalog number 1968-SL-050, 1 μg / mL in TSMBB), Siglec-3 (Fc-tagged, R&D system catalog number 1137-SL-050, 1 μg / mL in TSMBB), Siglec-5 (Fc-tagged, R&D system catalog number 1072-SL-050, 20 μg / mL in TSMBB), Siglec-6 (Fc-tagged, R&D system catalog number 2859-SL-050, 1 μg / mL in TSMBB), Siglec-7 (Fc-tagged, R&D system catalog number 1138-SL-050, 20, 10, and 51 μg / mL in TSMBB), or Siglec-9 (Fc-tagged, R&D system catalog number 1138-SL-050, 20, 10, and 51 μg / mL in TSMBB). After incubation with goat anti-human IgG antibody (Alexa Fluor 647-conjugated, Jackson ImmunoResearch, 10 μg / mL) for 1 hour, slides were washed and incubated for 30 minutes with a solution of goat anti-human IgG antibody (Alexa Fluor 647-conjugated, Jackson ImmunoResearch, 10 μg / mL). For screening of strep-tagged enterovirus proteins B3 / 1013, A2 / 2018, B2 / 039, A4 / P4, and H5VN and H3N8 hemagglutinins, slides were incubated with the proteins (50 μg / mL in TSMBB) for 1 hour, washed, and incubated for 30 minutes with a solution of anti-Strep tag antibody (StrepMAB-Classic Oyster 645, IBA Lifesciences 2-1555-050, 10 μg / mL). For screening of MERS spike protein, slides were incubated with a premixed mixture of MERS spike protein (Fc-tagged, 50 μg / mL) and pAb-LS nanoparticles (50 μg / mL) for 1 hour, then washed and incubated with a solution of goat anti-human IgG antibody (Alexa Fluor 647-conjugated, Jackson ImmunoResearch, 10 μg / mL) for 30 minutes. After washing and drying, slides were scanned at the appropriate excitation wavelengths with a GenePix 4000B microarray scanner (Molecular Devices) at a resolution of 5 μm.Various gain and PMT values ​​were used for the scans to ensure that all signals were within the linear range of the scanner's detector and that there was no signal saturation. Images were analyzed using GenePix Pro 7 software (version 7.2.29.2, Molecular Devices). Data were analyzed using an Excel macro (http: / / zenodo.org / record / 5146251). The highest and lowest values ​​of the total fluorescence intensity for each replicate were removed, and the remaining values ​​were used to calculate the mean and standard deviation.

[0496] The results are shown in Figures 3A-3M (the vertical axis corresponds to the fluorescence intensity measured in relative fluorescence units (RFU)).

[0497] Example 5: Surface plasmon resonance analysis of binding to Siglecs The K of the bond of the compounds of the present invention d is determined using surface plasmon resonance (SPR) technology.

[0498] Surface / Sample Conjugation: All analyses were performed on a GE Biacore T100 instrument. Streptavidin (2,000 response units, Thermo Fisher Scientific No. 434301) was coated onto a Series Scm5 chip (Cytiva No. 29149603) using standard EDC / NHS amide conjugation chemistry (flow cells 1, 2, 3, and 4). All flow cells were then blocked with ethanolamine, and the chip surface was stabilized by flowing (10 μL / min) overnight with a degassed solution of PBS-P (10 mM NaHPO, 1.8 mM KHPO, 137 mM NaCl, 2.7 mM KCl, and 0.5% surfactant P20 (Cytiva No. BR100054), pH 7.4).

[0499] Once the baseline has stabilized, introduce a biotinylated compound dissolved in DI water at a concentration of 50 μg / mL (total volume = 200 μL). Using the T100 software in manual mode, introduce a 50 μg / mL solution of biotinylated disialoside into flow cell 2 at a flow rate of 30 μL / min for 90 seconds, followed by PBS-P at a flow rate of 30 μL / min for 90 seconds. This process is repeated three times, after which no additional biotin conjugation is observed.

[0500] Siglec-3 screening Lyophilized hFc-Siglec-3 (R&D Systems No. 1137-SL-050) is reconstituted in PBS-P buffer to a concentration of 200 μg / mL and allowed to equilibrate on ice for 30 minutes before analysis. A binding assay is set up to measure the interaction between Siglec-3 and biotinylated sialoside by subtracting the response units measured from flow cells 2 and 1. This analysis is accomplished by introducing Siglec-3 at three different concentrations: 200 μg / mL, 100 μg / mL, and 50 μg / mL, using the following parameters: a. Flow rate = 30 μL / min; b. Contact time = 60 seconds; c. Dissociation time = 90 seconds; d. Stabilization time = 30 seconds.

[0501] Siglec-5 screening Lyophilized hFc-Siglec-5 (R&D Systems No. 1072-SL-050) is reconstituted in PBS-P buffer to a concentration of 200 μg / mL and allowed to equilibrate on ice for 30 minutes before analysis. A binding assay is set up to measure the interaction between Siglec-3 and biotinylated sialoside by subtracting the response units measured from flow cells 2 and 1. This analysis is accomplished by introducing Siglec-5 at three different concentrations: 200 μg / mL, 100 μg / mL, and 50 μg / mL, using the following parameters: a. Flow rate = 30 μL / min; b. Contact time = 60 seconds; c. Dissociation time = 90 seconds; d. Stabilization time = 30 seconds.

[0502] Siglec-7 screening Lyophilized hFc-Siglec-7 (R&D Systems No. 1138-SL-050) is reconstituted in PBS-P buffer to a concentration of 200 μg / mL and allowed to equilibrate on ice for 30 minutes before analysis. A binding assay is set up to measure the interaction between Siglec-7 and biotinylated sialoside by subtracting the response units measured from flow cells 2 and 1. This analysis is accomplished by introducing Siglec-7 at three different concentrations: 200 μg / mL, 100 μg / mL, and 50 μg / mL, using the following parameters: a. Flow rate = 30 μL / min; b. Contact time = 60 seconds; c. Dissociation time = 90 seconds; d. Stabilization time = 30 seconds.

[0503] Siglec-9 screening Lyophilized hFc-Siglec-9 (R&D Systems No. 1139-SL-050) was reconstituted in PBS-P buffer to a concentration of 200 μg / mL and equilibrated on ice for 30 minutes before analysis. A binding assay was set up to measure the interaction between Siglec-9 and biotinylated sialoside by subtracting the response units measured from flow cells 2 and 1. This analysis was accomplished by introducing Siglec-9 at concentrations of 100 μg / mL, 50 μg / mL, and 25 μg / mL using the following parameters: a. Flow rate = 30 μL / min; b. Contact time = 60 seconds; c. Dissoc...

Claims

1. The following structural formula: 【Chemistry 1】 (In the formula, R １ is, independently at each occurrence, -C(O)-A, where A is C １ ~C ６ Alkyl, C ６ ~C １８ Aryl, (C ６ ~C １８ ) aryl (C １ ~C ３ ) alkyl, 5- to 18-membered heteroaryl, (5- to 18-membered) heteroaryl (C １ ~C ３ ) alkyl, C ３ ~C ８ cycloalkyl, (C ３ ~C ８ ) cycloalkyl (C １ ~C ３ ) alkyl, 5- to 8-membered heterocycloalkyl or (5- to 8-membered) heterocycloalkyl(C １ ~C ３ ) alkyl; one or two carbon atoms within the alkyl portion of A are each independently optionally replaced with a heteroatom selected from N, O, or S; A is 1 to 3 R １１ groups, and each of the R １１ The group is C １ ~C ６ Alkyl, C １ ~C ６ Haloalkyl, C １ ~C ６ Alkoxy, halogen, C ６ ~C １２ or two groups R independently selected from aryl, 5- to 12-membered heteroaryl and cyano; １１ together with the atom to which they are attached form a 5-7 membered heterocyclyl having 1 to 3 heteroatoms selected from N, O or S; R １１ are each independently a halogen, C １ ~C ６ Alkyl, C １ ~C ６ Haloalkyl, C １ ~C ６ optionally substituted with 1 to 3 substituents selected from alkoxy or cyano; moreover, R is independently at each occurrence -R Ｌ -R Ｆ and R Ｌ represents independently at each occurrence -O-(C １ ~C １２ ) alkylenyl-, —O—, —S—, —NR １００ -, -S-(C １ ~C １２ ) alkylenyl-, -NR １０１ -(C １ ~C １２ ) alkylenyl-, -NR １０１ａ -O-(C １ ~C １２ ) alkylenyl-; —O—(CH ２ CH ２ O) ｍ -, -O-(CH ２ CH ２ O) ｋ - (CH ２ CH ２ ) -, -NR １０２ -X １００ -(C １ ~C １２ ) alkylenyl-, -NR １０２ａ -NR １０２ｂ -C(O)-(C １ ~C １２ ) alkylenyl, and R １００ , R １０１ , R １０１ａ , R １０２ , R １０２ａ and R １０２ｂ are each independently H or C １ ~C ３ alkyl, and X １００ is —O— or —NH—, and m and k each independently represent an integer of 1 to 12; R Ｆ is expressed independently for each occurrence as H, C １ ~C ３ Alkyl, —NH ２ , -NH-Fmoc, -NH-Boc, -NH-CBz, -NH-Troc, -NH-TFA, Mono(C １ ~C ３ ) alkylamino, di(C １ ~C ３ alkyl)amino; or -C(O)-R １０３ (In the formula, R １０３ is —H, —OH or (C １ ~C ３ ) alkyl); The following structural formula 【Chemistry 2】 -SH, which is a moiety represented by Click Chemistry Reagents is) or a pharmaceutically acceptable salt thereof.

2. The following structural formula: 【Transformation 3】 2. The compound of claim 1, represented by any one of: or a pharmaceutically acceptable salt thereof.

3. The click chemistry reagent is an azide, C ２ ~C ３ The compound of claim 1, comprising an alkyne, a tetrazine, a trans-cyclooctene, or a cyclooctyne.

4. The click chemistry reagent is an azide, C ２ ~C ３ Alkynes or alkynes having the following structural formula: 【Chemistry 4】 (In the formula, R ｃｏ is hydrogen or halogen; X is absent or O; and R ０ In each case, hydrogen, halogen, C １～８ Alkyl, C １～８ Alkoxy, C ６ ~C １２ Aryl, 5- to 8-membered heteroaryl, C ３～８ cycloalkyl or 3- to 8-membered heterocyclyl; any two or more R ０ groups, together with the atoms to which they are attached, optionally form an unsaturated, saturated, or aromatic 5- to 8-membered ring; and R ｔｄ is hydrogen, C １～８ Alkyl, C １～８ Alkoxy, C ６ ~C １２ Aryl, 5- to 8-membered heteroaryl, C ３～８ cycloalkyl or 3- to 8-membered heterocyclyl) 4. The compound of claim 3, wherein the compound is a moiety represented by any one of:

5. The click reagent is an azide, C ２ ~C ３ Alkynes or alkynes having the following structural formula: 【Transformation 5】 (In the formula, R ＊ is H or methyl, and R ＃ is independently at each occurrence H or C １ ~C ３ alkyl, and R Ｘ is —NH—C(O)O—) 4. The compound of claim 3, wherein the compound is any one of the moieties represented by:

6. R is independently at each occurrence -O-(CH ２ ) ｘ -NH ２ or -O-(CH ２ ) ｘ 2. The compound of claim 1, wherein x is -NH-Fmoc and x is, independently at each occurrence, an integer from 1 to 10.

7. 7. The compound of claim 6, wherein x is 5.

8. The cycloalkyl portion of moiety A is C ６ ~C ７ cycloalkyl; The heterocycloalkyl portion of moiety A is selected from 5-6 membered heterocycloalkyl having 1 or 2 heteroatoms selected from N, O, or S; The aryl portion of moiety A is selected from phenyl or naphthalenyl; and The heteroaryl portion of moiety A has the following structural formula: 【Transformation 6】 (In the formula, X １ , X ２ and X ３ are each independently NR Ｈ , O or S; R Ｈ is H or C １ ~C ３ alkyl) and further comprising: If present, each R １１ is cyano, halogen, phenyl, halophenyl, C １ ~C ６ Alkyl, C １ ~C ６ Haloalkyl, C １ ~C ３ or two groups R １１ The compound of any one of claims 1 to 7, wherein: together with the atoms to which they are attached form a [1,3]dioxolo group optionally substituted with one or two methyl or ethyl groups.

9. R １ may independently represent at each occurrence the following structural formula: 【Transformation 7】 and the wavy line represents the R １ A compound according to any one of claims 1 to 7, wherein the compound represents an attachment point of

10. Compounds S-101 to S-131 or D-101 to D-131 Table 1 Table 2 Table 3 Table 4 Table 5 Table 6 Table 7 and the moiety R is —O—(CH ２ ) ５ The compound according to any one of claims 1 to 7, which is -NH-Fmoc, -O-(CH 2 ) 5 -NH 2 or -O-(CH 2 ) 5 -N 3.

11. The following structural formula: G-L-P (In the formula, P is a biocompatible polymer; L is a covalent linker; and G has the following structural formula: 【Transformation 8】 (In the formula, symbol 【Chemistry 9】 represents the point of attachment to L; R １ is, independently at each occurrence, -C(O)-A, where A is C １ ~C ６ Alkyl, C ６ ~C １８ Aryl, (C ６ ~C １８ ) aryl (C １ ~C ３ ) alkyl, 5- to 18-membered heteroaryl, (5- to 18-membered) heteroaryl (C １ ~C ３ ) alkyl, C ３ ~C ８ cycloalkyl, (C ３ ~C ８ ) cycloalkyl (C １ ~C ３ ) alkyl, 5- to 8-membered heterocycloalkyl or (5- to 8-membered) heterocycloalkyl(C １ ~C ３ ) alkyl; one or two carbon atoms within the alkyl portion of A are each independently optionally replaced with a heteroatom selected from N, O, or S; A is 1 to 3 R １１ groups, and each of the R １１ The group is C １ ~C ６ Alkyl, C １ ~C ６ Haloalkyl, C １ ~C ６ Alkoxy, halogen, C ６ ~C １２ or two groups R independently selected from aryl, 5- to 12-membered heteroaryl and cyano; １１ together with the atom to which they are attached form a 5-7 membered heterocyclyl having 1 to 3 heteroatoms selected from N, O or S; R １１ are each independently a halogen, C １ ~C ６ Alkyl, C １ ~C ６ Haloalkyl, C １ ~C ６ optionally substituted with 1 to 3 substituents selected from alkoxy and cyano or a pharmaceutically acceptable salt thereof) A particle comprising a compound represented by:

12. The moiety G has the following structural formula: 【Chemistry 10】 12. The particle of claim 11, represented by any one of: or a pharmaceutically acceptable salt thereof.

13. The particle of claim 11, wherein the biocompatible polymer comprises a copolymer PLGA-PEG.

14. P has the following structural formula: 【Chemistry 11】 (wherein the symbol 【Chemistry 12】 represents the point of attachment of the polymer to the linker L, and y is an integer from 0 to 1000, x is an integer from 0 to 1000, and m is an integer from 0 to 450, provided that x and y are not simultaneously 0. The particle of claim 11 , represented by:

15. 15. The particle of claim 14, wherein y is an integer from 0 to 500, x is an integer from 0 to 500, and m is an integer from 0 to 250.

16. The particle of any one of claims 11 to 15, wherein the linker L comprises a moiety that is the product of a click chemistry reaction.

17. The linker L has the following structural formula: 【Chemistry 13】 【Chemistry 14】 (In the formula, R ＊ is H or methyl, and R ＃ is independently at each occurrence H or C １ ~C ３ alkyl, and R Ｘ is —NH—C(O)O—, and R ２００ is -H or C １ ~C ３ alkyl) and a portion represented by any one of the symbols 【Chemistry 15】 and each independently represent a point of attachment of the linker L to an additional moiety, P, or G.

18. The particle according to any one of claims 11 to 15, wherein P is PLGA(10k)-PEG(5k).

19. 16. The particle according to any one of claims 11 to 15, wherein the weight of G per unit weight of P (ligand density) is 10 to 75 μg / mg.

20. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 or the particles according to any one of claims 11 to 15, or a pharmaceutically acceptable salt thereof, in a pharmaceutically acceptable carrier.

21. The pharmaceutical composition of claim 20, for use in treating a disorder in a subject in need thereof, wherein the disorder is selected from cancer, ophthalmic disease, fibrosis, parasitic inflammation, fungal inflammation, viral inflammation, autoimmune inflammation, neurogenic inflammation, neurodegeneration, skin inflammation, renal inflammation, cardiovascular disease, gastrointestinal inflammation, or rheumatic disease.

22. The pharmaceutical composition of claim 20, for use in treating a disorder in a subject in need thereof, wherein the disorder is selected from breast cancer, non-small cell lung cancer (NSCLC), prostate cancer, colorectal cancer, melanoma, pancreatic cancer, myelofibrosis, diabetic retinopathy, idiopathic pulmonary fibrosis, hepatic fibrosis, sickle cell anemia, and acute respiratory distress syndrome (ARDS).

23. The following structural formula: G-L-P (In the formula, P is a biocompatible polymer; L is a covalent linker; and G has the following structural formula: 【Chemistry 16】 (In the formula, symbol 【Chemistry 17】 represents the point of attachment to L; R １ is, independently at each occurrence, -C(O)-A, where A is C １ ~C ６ Alkyl, C ６ ~C １８ Aryl, (C ６ ~C １８ ) Ara (C １ ~C ３ ) alkyl, 5- to 18-membered heteroaryl, (5- to 18-membered) heteroarylene (C １ ~C ３ ) alkyl, C ３ ~C ８ cycloalkyl, (C ３ ~C ８ ) cycloalkyl (C １ ~C ３ ) alkyl, 5- to 8-membered heterocycloalkyl or (5- to 8-membered) heterocycloalkyl(C １ ~C ３ ) alkyl; one or two carbon atoms within the alkyl portion of A are each independently optionally replaced with a heteroatom selected from N, O, or S; A is 1 to 3 R １１ groups, and each of the R １１ The group is C １ ~C ６ Alkyl, C １ ~C ６ Haloalkyl, C １ ~C ６ Alkoxy, halogen, C ６ ~C １２ aryl, 5- to 12-membered heteroaryl, cyano, or two groups R １１ together with the atom to which they are attached form a 5-7 membered heterocyclyl having 1 to 3 heteroatoms selected from N, O or S; R １１ are each independently a halogen, C １ ~C ６ Alkyl, C １ ~C ６ Haloalkyl, C １ ~C ６ optionally substituted with 1 to 3 substituents selected from alkoxy and cyano or a pharmaceutically acceptable salt thereof) 1. A method of making a particle comprising a molecule represented by Structural formula (I) G-R Ｆ１ (I) with a compound represented by structural formula (II) -) Ｆ２ (=) (In the formula, R Ｆ１ and R Ｆ２ are reactive moieties) and a compound represented by the moiety R Ｆ１ and R Ｆ２ under conditions suitable to react with each other, thereby forming said covalent linker -L- moiety.

24. R Ｆ１ and R Ｆ２ is a click reagent pair, and the condition is R Ｆ１ and R Ｆ２ and the linker L is suitable for a click reaction between R Ｆ１ and R Ｆ２ 24. The method of claim 23, comprising a moiety that is the product of a click chemistry reaction between

25. The following structural formula: [Chemistry 18] (In the formula, R is independently at each occurrence -R Ｌ -R Ｆ and R Ｌ represents independently at each occurrence -O-, -S-, -NR １００ -, -O-(C １ ~C １２ ) alkylenyl-, -S-(C １ ~C １２ ) alkylenyl-, -NR １０１ -(C １ ~C １２ ) alkylenyl-, -NR １０１ａ -O-(C １ ~C １２ ) alkylenyl-; —O—(CH ２ CH ２ O) ｍ -, -O-(CH ２ CH ２ O) ｋ - (CH ２ CH ２ ) -, -NR １０２ -X １００ -(C １ ~C １２ ) alkylenyl-, -NR １０２ａ -NR １０２ｂ -C(O)-(C １ ~C １２ ) alkylenyl, and R １００ , R １０１ , R １０１ａ , R １０２ , R １０２ａ and R １０２ｂ are each independently H or C １ ~C ３ alkyl, and X １００ is —O— or —NH—, and m and k each independently represent an integer of 1 to 12; R Ｆ is expressed independently for each occurrence as H, C １ ~C ３ Alkyl, —NH ２ , -NH-Fmoc, -NH-Boc, -NH-CBz, -NH-Troc, -NH-TFA, Mono(C １ ~C ３ ) alkylamino, di(C １ ~C ３ alkyl)amino; -C(O)-R １０３ (In the formula, R １０３ is —H, —OH or (C １ ~C ３ ) alkyl); The following structural formula 【Chemistry 19】 -SH, which is a moiety represented by Click Chemistry Reagents However, -R Ｌ -R Ｆ is not -OH) or a pharmaceutically acceptable salt thereof.

26. The following structural formula: G-L-P (In the formula, P is a biocompatible polymer; L is a covalent linker; and G has the following structural formula: 【Chemistry 20】 (In the formula, symbol 【Chemistry 21】 represents the point of attachment to L) or a pharmaceutically acceptable salt thereof) A particle comprising a compound represented by:

27. 27. The particle of claim 26, wherein the polymer comprises a block copolymer PLGA-PEG.

28. 28. A pharmaceutical composition comprising the compound of claim 25 or a pharmaceutically acceptable salt thereof, or the particle of claim 26 or 27 or a pharmaceutically acceptable salt thereof, for treating a disorder in a subject in need thereof, wherein the disorder is influenza.

29. The compound according to claim 10, wherein the variable R is any one of —O—(CH ２ ) ５ -NH ２ , —O—(CH ２ ) ５ -NH-Fmoc, -O-(CH ２ ) ５ -N ３ or -O-(CH ２ ) ５ -biotin, or a pharmaceutically acceptable salt thereof.

30. The structural formula: 【Chemistry 22】 【Chemistry 23】 【Chemistry 24】 wherein, independently for each occurrence: x is an integer from 90 to 140; y is an integer from 10 to 75; and m is an integer from 90 to 140. or a pharmaceutically acceptable salt thereof.

31. R is independently at each occurrence -O-(CH ２ ) ｘ -N ３ and x is, independently at each occurrence, an integer from 1 to 10.

32. 32. The compound of claim 31, wherein x is 5.

33. The additional portion of the linker is —O—(C １ ~C １２ 18. The particle of claim 17, wherein the alkyl group is .

34. (a) The biocompatible polymer comprises at least one of polyglycolic acid, poly(lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, poly(3-hydroxybutyric acid), polyethylene oxide, polyoxyethylene-polyoxypropylene block copolymer, poly(hydroxymethyl methacrylate), polyvinyl alcohol, poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, polysialic acid, and chitosan; (b) the biocompatible polymer comprises poly(L-lactic acid), poly(D-lactic acid), poly(D / L-lactic acid), copolymers thereof, or combinations thereof; or 27. The particle of any one of claims 11, 12, or 26, wherein (c) the biocompatible polymer comprises polyethylene oxide, polyoxyethylene-polyoxypropylene block copolymer, copolymers thereof, or combinations thereof.

35. x is an integer from 90 to 140; y is an integer from 10 to 75; and 15. The particle according to claim 14, wherein m is an integer from 90 to 140.

36. P has the following structural formula: 【Chemistry 25】 (wherein the symbol 【Chemistry 26】 represents the point of attachment of the polymer to the linker L, and y is an integer from 0 to 1000, x is an integer from 0 to 1000, and m is an integer from 0 to 450, provided that x and y are not simultaneously 0.

27. The particle of claim 26, represented by:

37. x is an integer from 90 to 140; y is an integer from 10 to 75; and 37. The particle according to claim 36, wherein m is an integer from 90 to 140.

38. acid-terminated PLGA (PLGA-COOH); Acid-terminated PLGA-PEG copolymers, where the acid moieties terminate the PEG blocks (PLGA-PEG-COOH); A PLGA-PEG copolymer, wherein the PEG block has the following structural formula: 【Chemistry 27】 or a PLGA-PEG copolymer (PLGA-PEG-DBCO) terminating in a moiety represented by: A PLGA-PEG copolymer, wherein the PEG block has the following structural formula: 【Chemistry 28】 PLGA-PEG copolymer (PLGA-PEG-NHS) terminated by a moiety represented by 27. The particle of claim 11, 12, or 26, further comprising one or more of:

39. a blend of 75 wt% PLGA-COOH and 25 wt% PLGA-PEG-DBCO; a blend of 75 wt% PLGA-COOH and 25 wt% PLGA-PEG-NHS; A blend of 90% by weight PLGA-PEG-COOH and 10% by weight PLGA-PEG-DBCO; and PLGA-PEG-NHS 39. The particle of claim 38, comprising at least one of:

40. 27. A pharmaceutical composition comprising the compound of claim 25 or the particle of claim 26, or a pharmaceutically acceptable salt thereof, in a pharmaceutically acceptable carrier.

41. 40. The particle of any one of claims 11 to 15, 33, 35 or 39, wherein the amount of G per unit weight of the particle is from 1 μg / mg to 1000 μg / mg.

42. 40. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7 or 31 to 32 or a particle according to any one of claims 11 to 15, 33, 35 or 39 for use in treating a disorder responsive to modulation of the activity of a Siglec receptor.

43. 43. The pharmaceutical composition of claim 42, wherein the disorder is selected from cancer, an immune-related disorder, and an inflammation-related disorder.

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